1/10000.Negative control was prepared by dilution of normal negative plasma to 1/100 with plasma-free dilution buffer.\",\n \"Once the antibody source is added to the antigen coated wells of the microtitre plate, the plate is incubated for 1-2 hrs at 37° C. The wells are then decanted and washed with the washing solution, as described above.\",\n \"To all wells 100 μL of Peroxidase-conjugated AffiniPure Goat Anti-Human IgG, Fcγ Fragment Specific (Jackson Immuno Research Laboratories, West Grove, Pa., USA) at a dilution of 1/5000 in assay dilution buffer is added and incubated at room temperature for 1 hr.\",\n \"All wells are then decanted again and washed with the washing solution.\",\n \"To all wells 125 μL of substrate solution [100 mM Sodium Acetate Trihydrate (pH 5.5), 0.0045% w/v hydrogen peroxide, 0.01% (w/v) 3,3′,5,5′-tetramethylbenzidine (Sigma-Aldrich, USA)] is then added.\",\n \"The substrate solution in the wells is then incubated for 15 minutes at room temperature protected from light.\",\n \"The reaction is then stopped by adding 100 μL of 2M sulfuric acid solution and the plate read with a 96-well plate reader/spectrophotometer at 450 nm and at 650 nm wavelength.\",\n \"A standard curve was plotted as linear optical density response (difference between the readings at 450 nm and at 650 nm wavelength) vs log-dose concentration.\",\n \"The standard curve can then be fitted with an iterative 4-parameter logistic analysis.\",\n \"Controls and samples are then interpolated using the logistic equation derived from the standard curve.\",\n \"The sample concentrations can then be corrected by dilution rate.\",\n \"ELISA and PRNT Results Correlated To make a VIG product, the plasma from donors and the final VIG product are assayed to determine the amounts of vaccinia virus specific antibodies.\",\n \"The number of antibodies specific to vaccinia virus can be determined by an ELISA assay or by the PRNT assay.\",\n \"The ELISA assay measures the amount of antibodies that bind to the vaccinia antigen while the PRNT assay is a functional assay that measures the amount of antibodies that actually neutralize the virus.\",\n \"It was previously assumed that the results of a PRNT assay were more accurate (Lublin-Tennenbaum et al., 1990, Viral Immunology 3: 19-25).\",\n \"Samples of a VIG were diluted to a specific concentration.\",\n \"Parallel samples were assayed by ELISA and PRNT, and compared to a standard VIG that was assigned a titre of 10,000 U/ml.\",\n \"A Bivariate Fit of log By Log graph (FIG.\",\n \"1) demonstrates the correlation between the results of the ELISA assay and the PRNT assay.\",\n \"Correlation coefficient of 0.98 indicates a very high level of correlation.\",\n \"Thus, the plasma and VIG may be screened by either ELISA or PRNT assay for the presence of vaccinia antibodies.\",\n \"Linear Fit log PRNT=−1.597741+1.2015617 Log ELISA Requirement to Vaccinate Plasma Donors The above-described ELISA assay was used to test plasma samples from 5 naïve donors (i.e.\",\n \"donors that have never been vaccinated for Smallpox).\",\n \"All the plasma samples tested from these naïve donors exhibited vaccinia-binding activities near the base line (less than 3 U/mL).\",\n \"In comparison, a plasma pool from donors vaccinated against Smallpox during childhood, but not subsequently revaccinated, had an activity of approximately 80 U/mL.\",\n \"These data suggest that a vaccinia immune globulin (VIG) must be manufactured from the plasma of donors that have been previously exposed to an Orthopoxvirus infection or an Orthopoxvirus vaccine.\",\n \"The plasma pool having an activity of approximately 80 U/mL was used to manufacture a Vaccinia Immune Globulin (VIG) solution (by the method described herein).\",\n \"The finished VIG product resulted in a 4.5% protein solution with an activity of around 800 U/mL, or approximately 8% of the activity of the VIG standard solution produced by Baxter.\",\n \"A VIG with 800 U/ml activity may be suitable for certain applications, such as for intravenous administration.\",\n \"The above results suggest that to produce a 5 g % (w/v) protein VIG comparable in activity to the VIG standard solution produced by Baxter (i.e.\",\n \"10,000 IU/ml), donor plasma having an activity of about 1,000 IU/ml should be collected.\",\n \"The above calculations are predicated on the assumption that the ELISA assay will correlate with the PRNT assay.\",\n \"As stated previously, a more informative assay is an assay that detects the levels of protective antibodies, such as the PRNT assay.\",\n \"In a preferred embodiment, a VIG should be produced wherein the levels of protective antibodies are comparable to the levels found in the standard solution produced by Baxter.\",\n \"However, the above results suggest that revaccination of the plasma donors with the Smallpox vaccine might be required to elevate the levels of vaccinia virus antibodies, and thus produce a VIG with increased potency.\",\n \"Immunization of Donors Donors were adults in generally good health, who had previously been immunized with vaccinia vaccine, as evidenced by a visible vaccination scare and either a written or oral history of being vaccinated.\",\n \"Dryvax®, the vaccinia (Smallpox) vaccine, available through the US Centers for Disease Control and Prevention in Atlanta, Ga., is a lyophilized, live-virus preparation of infectious vaccinia virus (Wyeth Laboratories, Inc., Marietta, Pa.).\",\n \"Vaccinia vaccine does not contain Smallpox (variola) virus.\",\n \"The vaccine had been prepared from calf lymph with a seed virus derived from the New York City Board of Health (NYCBOH) strain of vaccinia virus and has a minimum concentration of 108 pock-forming units (PFU)/ml.\",\n \"Vaccine was administered by using the multiple-puncture technique with a bifurcated needle, supplied with the vaccine.\",\n \"Typically, the site of vaccination is on the shoulder.\",\n \"No preparation of the skin at the site of injection is required.\",\n \"However, if site is obviously dirty, a cloth moistened with water may be used to wipe the site.\",\n \"Use of a disinfectant can kill the vaccine virus.\",\n \"Withdrawal of vaccine from ampoule.\",\n \"A sterile bifurcated needle (which is supplied with the vaccine and must be cool) is inserted into the ampoule of reconstituted vaccine.\",\n \"On withdrawal, a droplet of vaccine, sufficient for vaccination, is contained within the fork of the needle.\",\n \"Application of Vaccine to the Skin The needle is held at a 90 degree angle (perpendicular) to the skin.\",\n \"The needle then touches the skin to release the droplet of vaccine.\",\n \"For both primary and revaccination, 15 up and down (perpendicular) strokes of the needle are rapidly made in the area of about 5 mm in diameter (through the drop of vaccine deposited on the skin).\",\n \"The strokes should be sufficiently vigorous so that a trace of blood appears at the vaccination site.\",\n \"If a trace of blood does not appear, the strokes have not been sufficiently vigorous and the procedure should be repeated.\",\n \"Although it is desirable not to induce frank bleeding, the proportion of successful takes is not reduced if bleeding does occur.\",\n \"No dressing should be used after vaccination.\",\n \"Safety of Revaccinating Plasma Donors with Dryvax® Vaccine Since systematic vaccination for Smallpox ended in North America nearly 30 years ago, for the vast majority of the population, primary Smallpox vaccination occurred more than 30 years ago.\",\n \"To determine the safety of revaccinating with Smallpox vaccine 30 years after primary vaccination, 20 human study subjects with a previous history of Smallpox vaccination, were revaccinated with Dryvax® according to the procedure described above.\",\n \"The study subjects were then monitored for 8 weeks, and their observed reactions to the vaccine are summarized in Table 1.The typical “pox reaction” following vaccination with Dryvax® evolved with the following characteristics.\",\n \"1.Erythema surrounding the vaccination site can occur as early as 30 minutes following vaccination.\",\n \"In these 20 subjects erythema was seen in 100% of the cases, the mean duration of the erythema was 14 days and had a peak incidence of occurrence at day 7.2.Swelling around the vaccination site was observed in 95% of subjects, was most frequently reported on day 7 and lasted a mean of 10 days.\",\n \"3.Blistering (vesiculation) occurred in only 65% of the vaccinated subjects, lasted a mean of 5.5 days and occurred on day 2 most frequently.\",\n \"4.Pustulation of the vesicle occurred in 80% of subjects, peaked 9 days following vaccination and lasted a mean of 5 days.\",\n \"It is interesting to note that in 5 subjects, pustulation was reported without vesiculation.\",\n \"5.Crusting of the lesion occurred in 95% of subjects, was seen most frequently at day 14 and lasted a mean of 7 days.\",\n \"6.Pruritis of the vaccination site was reported 90% of the time, was seen most frequently at day 7 and had a mean duration of 8.5 days.\",\n \"During the study period, no unexpected adverse events were observed in these 20 subjects, and there were no serious vaccine complications that are reported in the literature such as accidental inoculation to other sites, eczema vaccinatum, vaccinia necrosum or vaccinia encephalitis.\",\n \"These results indicate the safety of revaccinating with Smallpox vaccine to stimulate antibody production in plasma donors.\",\n \"Efficacy of Revaccinating Plasma Donors on Vaccinia Virus Antibody Levels Following vaccination of the 20 human subjects described above, plasma was collected from the subjects by automated plasmaphoresis devices PCS®2 System by Haemonetics Corporation (Braintree, Mass.)\",\n \"and Autopheresis-C from Baxter (Round Lake, Ill.).\",\n \"The average unit collected is 0.8 liters.\",\n \"The plasma was screened for the presence of disease indications as required by the FDA, and the levels of vaccinia binding antibodies were determined by the ELISA assay described above.\",\n \"Table 2 summarizes the results.\",\n \"The results indicate that there was a significant increase in the levels of vaccinia-binding antibodies at 10 days post vaccination, and the elevated antibody levels persisted until approximately 60 days post vaccination.\",\n \"The vaccinia-binding antibody levels significantly decrease approximately 7 weeks post vaccination, though they remain elevated compared to pre-vaccination levels.\",\n \"This would suggest the optimal time to collect plasma from the donors for the preparation of VIG is during the period when the vaccinia binding antibody levels are high, which in this application is between 10 and 60 days post revaccination.\",\n \"All 20 of the subjects demonstrated a typical “pox reaction” following vaccination (i.e.\",\n \"the development of a pustular lesion at 6-10 days post vaccination).\",\n \"However, approximately 20% of the plasma donors demonstrated a very low antibody response to vaccination (less than 100 U/ml) as determined by the ELISA assay.\",\n \"This suggests that the current practice of observing the development of a pustular lesion at 6-10 days post vaccination is not an accurate indication of the “take” of a vaccine.\",\n \"Therefore, the plasma samples may be screened for vaccinia-binding antibody levels or for protective antibodies, prior to use of the plasma for the production of VIG.\",\n \"Such results also suggest that the effectiveness of vaccination should be determined using an assay to detect the levels of vaccinia binding antibodies or protective antibodies.\",\n \"Time Course For Vaccinia Antibodies in Donor Plasma Plasma donors were selected if they had medical documentation of a previous Smallpox vaccination or a Smallpox vaccination scar.\",\n \"The plasma donors were vaccinated and plasma was drawn twice weekly for nine weeks.\",\n \"The plasma was assayed for vaccinia antibodies by an ELISA assay, as shown in FIG.\",\n \"2.Re-Vaccinating the Plasma Donors Since the titres of vaccinia antibodies in the donor plasma, an attempt was made to revaccinate the plasma donors with the Smallpox vaccine.\",\n \"Plasma donors were revaccinated with the Smallpox vaccine at least 12 weeks after the original vaccination.\",\n \"Plasma samples were collected and assayed for vaccinia antibodies.\",\n \"The results are shown in FIG.\",\n \"3.There was no increase in the titre of vaccinia antibodies.\",\n \"Plasma from donors is periodically tested for the titre of vaccinia antibodies by ELISA by the method described above.\",\n \"In one embodiment, plasma units with vaccinia antibody levels below 200 U per ml is not used.\",\n \"Plasma units are pooled, and the pools tested for vaccinia DNA by PCR and may be tested for vaccinia antibody levels.\",\n \"Manufacture of VIG Plasma Pool Preparation The frozen plasma units are allowed to thaw at room temperature, and the plasma is transferred to the pooling tank.\",\n \"The pooled plasma is diluted in the tank with water suitable for injection into human patients (WFI) (2 parts plasma: 1 part WFI) and mixed.\",\n \"Dextran sulphate, dissolved in WFI, is added to the pooled plasma tank to a final concentration of 0.03% (w/w) and mixed.\",\n \"The plasma is then filtered through cellulose filter pads (Seitz 700 or equivalent).\",\n \"Anion-Exchange Chromatography The anion exchange chromatography column is prepared in a cold room (approximately 8° C.).\",\n \"DEAE-Sephadex™ A50 resin (Pharmacia) is allowed to swell in 0.025 M potassium phosphate buffer pH 7.5 for 2-24 hours in a chromatography column.\",\n \"The DEAE-Sephadex™ is then treated sequentially with 1 M NaOH and 0.08 M H3PO4.The column is equilibrated by flushing with phosphate buffer.\",\n \"The Dextran-Sulphate treated plasma is applied to the DEAE-Sephadex column.\",\n \"When all the plasma has been applied, the column is eluted with equilibrating buffer (0.025M phosphate, pH 7.5) at the same flow rate.\",\n \"The column effluent absorption at UV280 is monitored and the UV280 absorbing peak (IgG effluent) is collected.\",\n \"The collected IgG effluent is filtered through two 35 nm Planova filters in series, and collected.\",\n \"Ultrafiltration The IgG effluent collected from the DEAE-Sephadex column is concentrated to approximately 0.5× the original volume by ultrafiltration, using a Millipore Pellicon ultrafiltration system (or equivalent) with 10,000 MW cut-off polysulfone cassettes.\",\n \"Addition of Solvent/Detergent Solution Lipid enveloped viruses are inactivated by a solvent detergent method.\",\n \"tri-n-butyl phosphate (TNBP) and Triton X-100 are added to a final concentration of 0.3% TNBP: 1.0% Triton X-100 and the solution is mixed for 4-4.5 hours at 4° C. The TNBP and Triton X-100 is removed by reverse phase chromatography using a column packed with C-18 resin and eluted with WFI.\",\n \"The column effluent absorption at UV280 is monitored and the UV280 absorbing peak, containing the IgG, is collected.\",\n \"0.2 Micron Filtration and Dispensing The IgG is pH adjusted to 5.5 with 1 M H3PO4, diafiltered with >4 volumes of cooled WFI using a Millipore Pellicon ultrafiltration system (or equivalent) with 10,000 MW cut-off polysulfone cassettes.\",\n \"Maltose is added to the IgG from the diafiltration step to a final concentration of 10% w/w, and polysorbate 80 is added to a final concentration of 0.03% w/w and mixed.\",\n \"The formulated IgG solution is aseptically filtered through a 0.2 micron filter, stored at 2-8° C. until QC testing is complete and aseptically dispensed into vials.\",\n \"The vials are stoppered, sealed with aluminum caps and stored at 2-8° C. until ready for packaging.\",\n \"The immune globulin formulation thus produced (VIG-CAN-1, will be used for further studies as outlined below) QPCR Analysis of Plasma and VIG Product The target DNA sequence is a region of the B18R gene encoding the interferon-alpha/beta receptor from the vaccinia virus strain Wyeth (GenBank Accesssion # AJ269556).\",\n \"A set of forward and reverse primers were designed using Primer Express software version 2.0 from Applied Biosystems.\",\n \"The sequence of the forward primer is: 5′-TTCGGAGGCACAATGAATGA-3′ (SEQ ID NO.\",\n \"1), and anneals between residues 240 and 259.The sequence of the reverse primer is: 5′-TGTCTTCAATAGGAGGACACTTTGC-3′ (SEQ ID NO.\",\n \"2), and anneals between residues 312 and 288.A vaccinia DNA standard is a pUC119 plasmid encoding a 113 bp insert from residues 220 to 332 of the GenBank plasmid.\",\n \"A cloned internal control standard differs from the vaccinia DNA standard by a four base pair deletion (GAGC) from residues 276 to 279 and a four base pair insertion (AGCG) between residues 284 to 285.The sequence of the vaccinia MGB probe is: 5′-6-carboxyfluorescein (FAM)-CTCTAGGAGAGCCATTC-MGB-3′ (SEQ ID NO.\",\n \"3), anneals between residues 268 and 284 of the vaccinia DNA standard.\",\n \"The sequence of the internal control MGB probe is: 5′-VIC-CTCTAGGACATTCAGCG-MGB-3′ (SEQ ID NO.\",\n \"4).\",\n \"In the presence of the target and internal control sequences, two amplicons are produced that are 73 bp and 69 bp, respectively.\",\n \"The QPCR is analyzed on an ABI PRISM® 7700 Sequence Detection System (Applied Biosystems, Foster City, Calif.).\",\n \"Distinction is made between the Vaccinia target sequence and the internal control sequence due to a four base pair mismatch in the DNA sequence of the two Taqman MGB probes as well as the use of two different reporter dyes to label the target and the internal control, namely, the FAM signal for vaccinia target sequence detection and the VIC signal for internal control sequence detection.\",\n \"Simultaneous amplification with these two Taqman MGB probes results in a multiplex QPCR assay that provides simultaneous detection of the vaccinia target sequence and the internal control sequence in the same closed tube.\",\n \"The main advantage of using an internal control sequence designed with a different probe hybridization site is that it makes the QPCR assay suitable for quality control applications where co-detection of the target and the internal control is required in the prevention of false negative.\",\n \"A standard curve was prepared in a related sample matrix (i.e.\",\n \"plasma) extracted by QIAamp® UltraSens™ Virus Kit (Qiagen) and is included in every QPCR run.\",\n \"It consists of five concentrations of the DNA standard spanning a range above and below the expected copy number in the test samples: 106, 105, 104, 2500 and 250.The standard curve plot consists of the log of the starting copy number of the DNA standards versus the threshold cycle (Ct) value, which is a straight line.\",\n \"The Ct values are inversely proportional to the copy numbers values therefore the higher the copy number, the lower the Ct values.\",\n \"QPCR reaction mixtures, the final concentration of forward and reverse Vaccinia primers were 900 nM each.\",\n \"The final optimized concentration of the Vaccinia TaqMan MGB probe as well as the internal control TaqMan MGB probe was determined to be 225 nM each.\",\n \"Primers and probes were synthesized by Applied Biosystems.\",\n \"TaqMan Universal 2×Master Mix (Applied Biosystems) was used in all QPCR reactions.\",\n \"Twelve microlitres of the 30 μL total eluant was analyzed per well.\",\n \"All reaction volumes used per well were 50 μL.\",\n \"The universal cycling parameters used were according to Applied Biosystems recommendations.\",\n \"The Sequence Detection Software plots the standard curve and determines the slope, R value and the Y-intercept for the standard curve.\",\n \"A slope of −3.3 is indicative of 100% PCR efficiency whereby there is the expected 3.3 Ct difference based on a ten-fold dilution.\",\n \"The Y-intercept provides the PCR cycle that corresponds to the cycle where a single copy of the target can be detected.\",\n \"The PCR efficiency can be calculated using the formula: 10−1/S−1 where S is the slope of the standard curve.\",\n \"The copy number in the unknowns is calculated by interpolation from the standard curve using the slope and the Y-intercept values.\",\n \"Experiments with known numbers of live vaccinia virus strain Wyeth, added to 500 μL of vaccinia-negative plasma and DNA extracted by QIAamp® UltraSens™ Virus Kit (Qiagen) according to manufacturers instructions, proved the efficacy of the method in detecting vaccinia virus.\",\n \"Ten VIG plasma mini-pools, a Cangene VIG bulk drug product (lot # 196.01.001), a Cangene VIG final product (lot # 0730203) were tested by the QPCR method.\",\n \"The standard curve consisted of the following points, 106, 105, 104, 2500 and 250 copies resulting in Ct values of 17.54, 20.69, 23.73, 25.71 and 26.80 respectively.\",\n \"The standard curve had an R value of 0.944.The Ct values for the samples tested were greater than 30.Such results indicate all ten VIG plasma mini-pools, Cangene VIG bulk drug product and Cangene VIG final product to have less than 1250 copies of vaccinia virus genomes per milliliter.\",\n \"Clinical Studies Safety Study The safety and pharmacokinetic characteristics of VIG-CAN-1 given in two intravenous doses will be determined.\",\n \"The VIG-CAN-1 is drawn up with an equivalent amount of Saline USP for injection such as the total volume of the solution is to equal to 0.6 mL/kg: 0.3 ml/kg of VIG-CAN-1 intravenously (n=25) with 0.3 ml/kg Saline USP or 0.6 ml/kg of VIG-CAN-1 intravenously (n=25) The VIG-CAN-1 will be given intravenously via appropriate canula.\",\n \"After administration of the VIG-CAN-1, blood samples will be collected and adverse events recorded (at 30 min., 1, 2, 4, 8 and 24 hours).\",\n \"Subjects return over the course of six weeks on days 1, 3, 5, 7, 14, 21, 28, 42, to provide additional blood and urine samples for analysis, and for adverse event and concomitant medication reporting.\",\n \"A complete physical examination will be performed at screening and day 42.Study Assessments For the preparation of samples, 5 mL of blood is drawn into sterile, pyrogen-free, vacuum blood collection tubes using sodium heparin (15 U/ml blood) as anticoagulant.\",\n \"Two 5 mL samples of plasma to be measured for vaccinia IgG levels by ELISA assay will be sent at the following times after study drug administration: Time 0, (before administration of VIG-CAN-1) 30 min, 1 hr, 2 hr, 4 hr, 8 hr, 24 hr Day 1 (same as 24 hr sample), 3, 5, 7, 14, 21, 28, 42 The following tests are conducted at Baseline, days 7, 28, and 42: Hematology: including RBC, Hgb, Het, WBC, Differnetial cell counts, Platelet count, RBC indices, Prothrombin time, Partial thromboplastin time, Reticulocyte, Fibrinogen, ESR Blood Chemistry calcium, Urea nitrogen, Glucose [random], Total Bilirubin, AST, ALT, Alkaline Phosphatase, Lactate Dehydrogenase, Creatinine, Sodium, Potassium, Chloride, Albumin, Total proteins Urinalysis (Specific Gravity, pH, Protein, Glucose, Ketones, Bilirubin, and blood) A full physical examination will be conducted at the 28th and 42nd day visit The study subjects are questioned during all assessment visits following drug administration about adverse events they may be experiencing and concomitant medications they may be taking Beta hCG [for premenopausal female subjects] Assessments for Safety An adverse event (AE) is any untoward medical occurrence in a patient or clinical investigation subject.\",\n \"An AE includes significant exacerbation of any baseline medical condition including, but not limited to, the disease under study.\",\n \"The event must occur in association with drug treatment but a causal relationship to treatment is not necessarily proven or implied.\",\n \"Data collected to date indicates intravenous injection of the VIG-CAN-1 at 6,000 U/Kg is safe in humans, with no adverse events reported at the 6-week follow-up visit.\",\n \"Preclinical Studies Mouse Tail Infectivity Model: VIG has been indicated for prevention or modification of aberrant infections induced by the vaccinia virus vaccine.\",\n \"The following protocols employ a mouse tail infectivity model that involves inoculation of vaccinia virus into the murine tail vein.\",\n \"This model of infectivity closely resembles the human vaccination process in that dermal lesions occur on the tail similar to the vaccination site lesion that occurs in humans, and both result in a self-limiting infection.\",\n \"Therefore, this is a good surrogate model for VIG-CAN-1 efficacy testing with this indication.\",\n \"A tail infectivity model for vaccinia is established using female Balb/c mice (Weight range 19 to 23 g) challenged with Vaccinia strain IHD-J administered by tail vein injection.\",\n \"These animals are challenged with 1×105 PFU of virus in 200 μl total volume and the pox appearing on the tail scored for 7 days.\",\n \"The dose of virus leading to 50 pox per tail are determined and this dose will be used for subsequent challenge studies with vaccinia immune globulin (VIG-CAN-1).\",\n \"The post exposure protection of VIG-CAN-1 is determined in the above described tail infectivity model.\",\n \"VIG-CAN-1 is tested in Balb/c mice challenged with a dose for vaccinia virus determined above to result in 50 pox per tail.\",\n \"Five doses of test VIG-CAN-1 is administered 24 hours after infection.\",\n \"After 7 days, the number of tail pocks is assessed by tail staining.\",\n \"The primary endpoint is the number of pocks per tail at Day 7.A dose response analysis is applied to the negative control, the five test VIG-CAN-1 groups and the five control VIG-CAN-1 groups.\",\n \"Positive control is Cidofovir™ administered intramuscularly at 100 mg/Kg in a total volume of 100 μl.\",\n \"Negative control is Saline administered at 100 μl intramuscularly.\",\n \"VIG-CAN-1 administered at 6000 U/Kg and 30000 U/KG intramuscularly.\",\n \"VIG-CAN-1 (30000 U/Kg) and Cidofovir™ (100 mg/Kg) administered intramuscularly in a total volume of 100 μl.\",\n \"Treatment with 6000 U/Kg of VIG resulted in similar number of pocks compared to the positive control.\",\n \"Vig 30000 and Vig 30000+ Cidofovir™ gave significantly fewer pocks compared to the positive control, as shown in Table 2.While the present invention has been described with reference to what are presently considered to be the preferred examples, it is to be understood that the invention is not limited to the disclosed examples.\",\n \"To the contrary, the invention is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.\",\n \"All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety.\",\n \"TABLE 1 Number of subjects Incidence Mean Peak Adverse Events (n = 20) (n = 20) Duration Occurrence Redness 20 100% 14 days Day 7 Swelling 19 95% 10 days Day 7 Crusting 19 95% 7 days Day 14 Itching 18 90% 8.5 days Day 7 Pustule 16 80% 5 days Day 9 Blistering 13 65% 5.5 days Day 2 Pain 10 50% 3 days Day 7 Headache 8 40% 1 day Day 2 URI 6 30% 3 days Day 2 Adhesive skin 5 25% 2 days Day 8 reaction Fatigue 4 20% 7 days Day 7-9 Other 4 20% 7 days Day 10 Discharge at 3 15% 9 days Day 9 vaccination site Myalgia 3 15% 3 days Day 11 Nausea 2 10% 2 days Day 7 or 9 Lymphadenopathy 2 10% 5 days Day 3 Fever 1 5% 1 day Day 2 Local Spread 0 0% Distant Lesions 0 0% Necrosis 0 0% Vomiting 0 0% TABLE 2 Compared to positive control Compared to negative control p value for Mean 95% Cl for Mean 95% Cl for difference with Treatment Mean 95% Cl difference difference difference difference positive control Saline 41.9 (36.7, 47.1) Cidofovir ™ 24.2 (21.0, 27.4) −17.7 (−23.3, −12.1) VIG 3000 23.7 (21.4, 26.0) −18.2 (−23.5, −12.9) −0.5 (−4.2, 3.2) p = 0.78 VIG 6000 26.5 (20.0, 33.0) −15.4 (−23.1, −7.7) 2.3 (−4.4, 9.0) p = 0.48 VIG 30000 16.9 (14.3-19.5) −27.3 (−31.9, −22.7) −7.7 (−11.5, −3.9) p < 0.001 VIG 30000 + 13.7 (11.0-16.4) −30.5 (−35.1, −25.9) −10.9 (−14.7, −7.1) p < 0.001 Cidofovir ™\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476853"}}},{"rowIdx":977,"cells":{"text":{"kind":"list like","value":[["Peptide-immobilized substrate and method for measuring target protein","Disclosed is a peptide-immobilized substrate for measuring a target protein, with which the peptide can have a structure required for being recognized by the target protein, with which the accurate loading amount of the peptide can be attained, and by which a trace amount of the target protein may be measured accurately and simply.","The peptide-immobilized substrate for measuring a target protein according to the present invention comprises a chemically synthesized peptide having an.","expected spatial structure or having a binding ability with the target protein, which peptide can bind with the target protein and is immobilized on the substrate."],["1.A peptide-immobilized substrate for measuring (a) target protein(s), comprising (a) chemically synthesized peptide(s) having (an) expected spatial structure(s) or having (a) binding ability (abilities) with said target protein(s), which peptide(s) can bind with said target protein(s) and which is(are) immobilized on said substrate.","2.The peptide-immobilized substrate according to claim 1, wherein said peptide(s) is(are) labeled with (a) label(s) whose signal(s) is(are) changed upon binding of said peptide(s) with said target protein(s).","3.The peptide-immobilized substrate according to claim 1, wherein said peptide(s) has(have) (an) amino acid sequence(s) which is(are) so molecularly designed as to attain said expected spatial structure(s).","4.The peptide-immobilized substrate according to claim 1, wherein the number of amino acids constituting said peptide is 3 to 50.5.The peptide-immobilized substrate according to claim 2, wherein said label(s) is(are) (a) fluorescent label(s) bound to (an) amino acid(s) located at (a) particular site(s), which amino acid(s) is(are) among the amino acids constituting said peptide.","6.The peptide-immobilized substrate according to claim 5, wherein two types of fluorescent labels are bound to particular amino acids located at different sites.","7.The peptide-immobilized substrate according to claim 2, wherein said label(s) is(are) bound to one or both terminal regions(s) of said peptide.","8.The peptide-immobilized substrate according to claim 1, wherein said peptide(s) is(are) one(those) synthesized while protecting functional groups in the side chains of the amino acids, and is(are) bound to said substrate at (a) desired site(s) by binding the peptide(s) to the substrate while protecting said functional groups which are then deprotected after said peptide(s) is(are) bound to said substrate.","9.The peptide-immobilized substrate according to claim 1, wherein the each peptide has (a) functional group(s) at its terminal region, which chemically binds to said substrate, and is bound to said substrate via said functional group.","10.The peptide-immobilized substrate according to claim 9, wherein the each peptide has cysteine at one end thereof, and is bound to said substrate through the sulfhydryl group of said cysteine.","11.The peptide-immobilized substrate according to claim 1, wherein the each peptide is bound to said substrate by covalent bond.","12.The peptide-immobilized substrate according to claim 1, wherein the each peptide has a core region to be subjected to the binding with said target protein, and (a) binding region(s) to be subjected to the binding to said substrate and to the binding with said label(s) in cases where said peptide has said label(s), said core region having spatial structure optimum to the binding with said target protein.","13.The peptide-immobilized substrate according to claim 1, wherein a plurality of said peptides are bound to prescribed sites on a single substrate.","14.A method for measuring (a) target protein(s) comprising contacting said peptide-immobilized substrate according claim 1 with said target protein(s); and measuring said target protein(s) bound to said peptide(s) based on the change of said signal(s).","15.A method for measuring (a) protein(s) in a test sample, comprising the steps of bringing a plurality of chemically synthesized peptides each of which has an expected spatial structure or a binding ability with the target protein, which can or might bind to the target protein, into contact with said test sample which may contain the target protein(s), said plurality of peptides being immobilized on the same or different substrates; measuring the signals which change upon binding between the peptides and the target protein(s); converting each of the values measured for each peptide into data with which differences between measured values may be recognized with naked eye and outputting visible data of the peptides in an array.","16.The method according to claim 15, wherein the visible physical value is color, brightness and/or chroma, and differences in color may be recognized with naked eye depending on the measured value for each peptide.","17.The method according to claim 15, wherein a plurality of chemically synthesized peptides which bind with a plurality of target proteins are immobilized.","18.A method for identifying or characterizing (a) protein(s) in a test sample comprising carrying out the method according to claim 15 for various test samples; accumulating outputted measured values to produce a database; comparing measured data for an unknown test sample with the measured data in said database thereby identifying or characterizing said unknown test sample.","19.The method according to claim 15, wherein at least a portion of said plurality of peptides contain random sequences in at least a part of their amino acid sequences.","20.The method according to claim 15, wherein at least a portion of said plurality of chemically synthesized peptides are labeled with labels whose signals are changed upon binding of said peptides with said target protein(s).","21.The method according to claim 15, wherein at least a portion of said plurality of chemically synthesized peptides have amino acid sequences which are so molecularly designed as to attain said expected spatial structure.","22.The method according to claim 15, wherein the number of amino acids constituting each of said plurality of chemically synthesized peptides is 3 to 50.23.The method according to claim 15, wherein each of said labels is a fluorescent label bound to an amino acid located at a particular site, which amino acid is one of the amino acids constituting said peptide.","24.The method according to claim 23, wherein two types of fluorescent labels are bound to particular amino acids located at different sites in a single peptide.","25.The method according to claim 24, wherein said labels are bound to one or both termini of said single peptide.","26.The method according to claim 15, wherein each of said chemically synthesized peptides is one synthesized while protecting functional groups in the side chains of the amino acids, and is bound to said substrate at a desired site by binding the peptide to the substrate while protecting said functional groups which are then deprotected after said peptide is bound to said substrate.","27.The method according to claim 15, wherein each of said chemically synthesized peptides has (a) functional group(s) at its terminus, which chemically binds to said substrate, and is bound to said substrate via said functional group.","28.The method according to claim 27, wherein each of said chemically synthesized peptides has cysteine at one terminus thereof, and is bound to said substrate through the sulfhydryl group of said cysteine.","29.The method according to claim 15, wherein said plurality of chemically synthesized peptides are bound to said substrate by covalent bond.","30.The method according to claim 15, wherein each of at least a portion of said plurality of chemically synthesized peptides has a core region to be subjected to the binding with said target protein, and (a) binding region(s) to be subjected to the binding to said substrate and subjected to the binding with said label(s) in cases where said peptide has said label(s), said core region having spatial structure optimum to the binding with said target protein."],["<SOH> BACKGROUND ART <EOH>Analyses of functions of proteins existing in the body are important tasks.","Methods for identifying and quantifying these expressed proteins satisfy the wide social demands from the clarification of cell functions in the research field to the diagnoses and therapies in the medical field.","As means for identifying and quantifying proteins, methods utilizing antibodies and various biosensor systems utilizing arrays of specific proteins are known.","The concept for developing the conventional protein chips is based on immobilization of proteins or fragments thereof, whose structures are known.","Immobilization of antibody molecules is also based on this concept.","By this method, quantification of the loading amount is problematic, and uniform immobilization is difficult.","Particularly, since the yield of immobilization (loading amount) is generally low and depends on the properties of the peptide fragments, the method lacks reliability and so wide use thereof is prevented.","Further, in cases where protein fragments are used, the sites of cleavage are restricted because of specificities of the cleavage (if the proteins are cleaved without specificities, the resultants are random fragments and so the molecules immobilized on the chips cannot be identified), thus, free design can not be accomplished.","Still further, the thus immobilized proteins have structures different from those in the body, and may not have the structures indispensable for attaining the desired recognitions.","These are problems in practice.","Methods for analyzing interactions between peptides and proteins in solutions using fluorometer or the like are known.","However, with these methods, considerable amount of samples are necessary, so that a long time is required for sample preparation.","Recently, methods such as surface plasmon or the like are used.","However, the apparatuses are expensive, the sensitivities are generally low, and simple measurements at local places cannot be attained.","Thus, by the conventional methods, treatments of numerous samples are difficult."],["<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG.","1 schematically shows the concept of the peptide-immobilized substrate according to the present invention.","FIG.","2 schematically shows a linear structure of the peptide used in the present invention.","FIG.","3 schematically shows the measurement carried out in Example 1 of the present invention.","FIG.","4 schematically shows the measurement carried out in Example 2 of the present invention.","detailed-description description=\"Detailed Description\" end=\"lead\"?"],["TECHNICAL FIELD The present invention relates to a peptide-immobilized substrate for measuring (a) target protein(s), and to a method for measuring the target protein(s) using the same.","BACKGROUND ART Analyses of functions of proteins existing in the body are important tasks.","Methods for identifying and quantifying these expressed proteins satisfy the wide social demands from the clarification of cell functions in the research field to the diagnoses and therapies in the medical field.","As means for identifying and quantifying proteins, methods utilizing antibodies and various biosensor systems utilizing arrays of specific proteins are known.","The concept for developing the conventional protein chips is based on immobilization of proteins or fragments thereof, whose structures are known.","Immobilization of antibody molecules is also based on this concept.","By this method, quantification of the loading amount is problematic, and uniform immobilization is difficult.","Particularly, since the yield of immobilization (loading amount) is generally low and depends on the properties of the peptide fragments, the method lacks reliability and so wide use thereof is prevented.","Further, in cases where protein fragments are used, the sites of cleavage are restricted because of specificities of the cleavage (if the proteins are cleaved without specificities, the resultants are random fragments and so the molecules immobilized on the chips cannot be identified), thus, free design can not be accomplished.","Still further, the thus immobilized proteins have structures different from those in the body, and may not have the structures indispensable for attaining the desired recognitions.","These are problems in practice.","Methods for analyzing interactions between peptides and proteins in solutions using fluorometer or the like are known.","However, with these methods, considerable amount of samples are necessary, so that a long time is required for sample preparation.","Recently, methods such as surface plasmon or the like are used.","However, the apparatuses are expensive, the sensitivities are generally low, and simple measurements at local places cannot be attained.","Thus, by the conventional methods, treatments of numerous samples are difficult.","DISCLOSURE OF THE INVENTION An object of the present invention is to provide a peptide-immobilized substrate for measuring (a) target protein(s), with which the peptide can have a structure required for being recognized by the target protein, with which the accurate loading amount of the peptide can be attained, and by which (a) trace amount(s) of the target protein(s) may be measured accurately and simply.","The present inventors intensively studied to discover that the above-described object may be attained by a peptide-immobilized substrate for measuring a target protein, comprising a chemically synthesized peptide having an expected spatial structures or having a binding ability with the target protein, which the peptide can bind with the target proteins and which is immobilized on the substrate, thereby completing the present invention.","Further, the present inventors inferred that a protein in a test sample may be easily and simply measured, or a protein contained in an unknown test sample may be identified or characterized by immobilizing a plurality of chemically synthesized peptides on a substrate; reacting the peptides with a test sample which may contain the target protein; assigning a visible physical value to the signal measured for each of the peptides; and outputting visible data of the peptides in an array.","That is, the present invention provides a peptide-immobilized substrate for measuring (a) target protein(s), comprising (a) chemically synthesized peptide(s) having (an) expected spatial structure(s) or having (a) binding ability(abilities) with said target protein(s), which peptide(s) can bind with said target protein(s) and which is(are) immobilized on said substrate.","The present invention also provides a method for measuring (a) target protein(s) comprising contacting said peptide-immobilized substrate according to the present invention with said target protein(s); and measuring said target protein(s) bound to said peptide(s) based on the change of said signal(s).","The present invention further provides a method for measuring (a) protein(s) in a test sample, comprising the steps of bringing a plurality of chemically synthesized peptides each of which has an expected spatial structure or a binding ability with the target protein, which can or might bind to the target protein, into contact with said test sample which may contain the target protein(s), said plurality of peptides being immobilized on the same or different substrates; measuring the signals which change upon binding between the peptides and the target protein(s); converting each of the values measured for each peptide into data with which differences between measured values may be recognized with naked eye and outputting visible data of the peptides in an array.","The present invention still further provides a method for identifying or characterizing (a) protein(s) in a test sample comprising carrying out the method according to the present invention for various test samples; accumulating outputted measured values to produce a database; comparing measured data for an unknown test sample with the measured data in said database thereby identifying or characterizing said unknown test sample.","By the present invention, a novel peptide-immobilized substrate for measuring a target protein, by which a trace amount of target protein may be measured accurately and simply, was provided.","In the peptide-immobilized substrate according to the present invention, each of the peptides is a chemically synthesized peptide, and may be immobilized on the substrate under the conditions -wherein the functional groups in the side chains of the amino acids are protected or unprotected.","That is, the peptide may be immobilized on the substrate at a particular site, and the effective immobilizing amount may be arbitrarily controlled.","Further, the peptide may be freely designed, so that an arbitrary label or functional molecular group may be attached to an arbitrary site, or a peptide containing an amino acid which does not occur in natural proteins may be employed.","Still further, by the fingerprint method according to the present invention, identification or characterization of the protein contained in the test sample may be attained easily and quickly, and a number of test samples or unknown test samples may be measured.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 schematically shows the concept of the peptide-immobilized substrate according to the present invention.","FIG.","2 schematically shows a linear structure of the peptide used in the present invention.","FIG.","3 schematically shows the measurement carried out in Example 1 of the present invention.","FIG.","4 schematically shows the measurement carried out in Example 2 of the present invention.","BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, the peptide to be immobilized on the substrate is a chemically synthesized peptide.","Most of the naturally occurring proteins have the properties to molecularly recognize the oligopeptides having particular spatial structures and bind with them (Reference: W. E. Stites, Chem Rev, 97, 1233-1250 (1997), Protein-protein interactions: Interface structure, binding thermodynamics, and mutational analysis; D. P. Fairlie et al, Curr.","Medicinal Chem., 5, 29-62 (1998) Toward protein surface mimetics; and A. G. Cochran, Chem & Biol, 7, R85-R94 (2000), Antagonist of protein-protein interactions).","It is predetermined what protein binds with what oligopeptide having what spatial structure.","The first candidate of the peptide used in the present invention is such a peptide having a particular spatial structure.","The spatial structures of the oligopeptide, which bind with proteins include various spatial structures such as α-helix, α-loop-α, β-turn, β-loop-β and β-strand.","As the peptide to be immobilized on the substrate, peptides having such a particular spatial structure are exemplified.","These spatial structures are those which the peptide may have when the peptide is bound with the target protein, and the peptide does not necessarily have these spatial structures when it is merely immobilized on the substrate.","In the present invention, the term “having an expected spatial structure” means that the peptide has the expected spatial structure when bound with the target protein.","It should be noted that the entire peptide does not necessarily has the expected spatial structure, as long as most part of the peptide (preferably not less than 60%, more preferably not less than 70% in terms of the number of amino acids) has the expected spatial structure.","Therefore, to one or both terminal regions of the peptide, the label(s) hereinbelow described may be bound, and the region(s) may not participate in the formation of the expected spatial structure.","It is preferred rather to separate the region(s) (hereinafter also referred to as “binding region”) at which the peptide is immobilized on the substrate or to which the label(s) is(are) attached and the region to be subjected to the binding with the target protein.","It is preferred here that the region (hereinafter also referred to as “core region”) to be subjected to the binding with the target protein have the spatial structure which is optimum to the binding.","In this case, the binding region(s) is(are) preferably attached to one or both termini of the core region.","The peptide having the above-mentioned spatial structure may have a circular structure.","In some cases, by making the peptide in a circular form, the binding affinity with the protein may be largely increased.","Although a peptide antigen may be chemically synthesized and may be used as the peptide for measuring an antibody, the binding between the peptide and the target protein is not necessarily antigen-antibody reaction at all.","As the proteins which bind with the oligopeptides having the above-described spatial structures, the following proteins are exemplified.","As the proteins which bind with an oligopeptide having α-helix structure, calmodulin and gp41 of HIV are exemplified.","As the proteins which bind with an oligopeptide having β-loop structure, amylase is exemplified.","As the proteins which bind with a β-strand peptide, MHC proteins, aspartic acid proteases, HIV-1 protease, cysteine proteases, metalloproteases and serine proteases are exemplified.","As the proteins which bind with a β-loop-β or β-turn peptide, fibronectin, LH-RH receptors, substance P receptors, somatostatin receptors and the like are exemplified.","But the proteins are not restricted thereto.","In addition to the above-described peptides having the expected spatial structures, peptide derivatives having binding abilities with the target proteins, such as those in which an organic molecule group (functional molecule group) known to have an interaction with a protein is attached to a particular site, may also be used as the peptide.","Examples of such functional molecule groups include porphyrin derivatives (heme), flavin derivatives, alkyl groups such as retinal and palmityl group, nucleic acid bases and peptide nucleic acids, but the functional molecule groups are not restricted thereto.","As the proteins which bind with these functional molecule groups, the following proteins are exemplified.","As the proteins which bind with porphyrin derivative (heme)-containing peptides, apo-heme proteins are exemplified.","As the proteins which bind with flavin derivative-containing peptides, apo-flavin proteins are exemplified.","As the proteins which bind with peptides containing alkyl groups such as retinal and palmityl group, lipocalin proteins are exemplified.","As the proteins which bind with the nucleic acid base-containing peptides, DNases, RNases, various gene function-controlling proteins and RNA-binding proteins are exemplified.","The concept of the peptide-immobilized substrate comprising a chemically synthesized peptide having an expected spatial structure or having a binding ability with the target protein, which peptide is immobilized on the substrate, is schematically shown in FIG.","1.With the technology at present, what amino acid sequence is necessary for attaining a particular spatial structure may be easily determined by using a computer.","Computer softwares for carrying out such a molecular designing are commercially available, and the sequence of the peptide having the expected spatial structure may easily be determined by using the computer softwares.","The size of the peptide is not restricted, and usually, the number of amino acids is preferably about 2 to 50, more preferably about 5 to 20.The target protein bound with the peptide immobilized on the substrate may be measured by various methods.","Examples of these methods include the methods in which the peptide to be immobilized is labeled with (a) label(s) whose signal changes upon binding of the peptide with the target protein, and the target protein is measured based on the change of the label(s); surface plasmon resonance method; Grating method which is a modification of surface plasmon that does not use a prism; and mass-spectrometric analyses, but the methods are not restricted thereto.","Among these, the methods in which the peptide to be immobilized is labeled with (a) label(s) whose signal changes upon binding of the peptide with the target protein are preferred because they have the highest sensitivities and reproducibilities.","As mentioned above, the peptide used in the present invention is preferably labeled with (a) label(s) which changes the signal upon binding of the peptide with the target protein.","Examples of such a label include fluorescent labels, spin labels, UV or visible light-absorbing dyes, radioisotopes, stable isotopes and enzymes which cause chemiluminescent reactions (alkaline phosphatase, acid phosphatase, luciferase, green fluorescence protein and the like), but the labels are not restricted thereto.","Among these, fluorescent labels are most preferred.","Examples of fluorescent labels include fluorescein and derivatives thereof having the fluorescein structure such as carboxyfluorescein; dansyl group and derivatives thereof having the structure of dansyl group such as ethyldansyl group; coumarin derivatives; pyrene derivatives; and naphthalene derivatives, but the fluorescent labels are not restricted thereto.","These fluorescent label reagents per se are known and many of them are commercially available.","Further, the methods per se for labeling an amino acid by binding a commercially available fluorescent dye with (a) functional group(s) in the amino acid, such as amino group, are known.","For example, the label may easily be introduced by binding the label to the side chain of lysine, glutamic acid or cysteine, or to the amino terminal or carboxyl terminal of the peptide, by the method described in Example 1.Since various fluorescence-labeled building blocks used for the synthesis are commercially available from Molecular Probe, U.S. and so on, these commercially available building blocks may be used as they are.","As preferred examples of the fluorescent labels, chemical structures of tetramethylrhodamine and carboxyfluorescein are shown in Formulae [I] and [II] below, respectively.","The signals emitted by the above-described labels are changed upon binding of the peptide with the target protein.","Although the signal is changed even in cases where a single label is bound to the peptide, it is preferred to bind a plurality of different labels to different sites in the peptide because the change of the signals may be detected more sensitively and so the detection sensitivity may be further promoted.","In the especially preferred mode, two types of fluorescent pigments whose fluorescent wavelengths are different are bound to different sites in the peptide.","By so doing, sensitive detection may be attained utilizing fluorescence resonance energy transfer (FRET) system.","Examples of the combinations of the fluorescent labels, which are preferred for attaining FRET, include combinations of a fluorescein derivative and a rhodamine derivative; a dansyl derivative and a fluorescein derivative; and a pyrene derivative and a fluorescein derivative, but the combinations are not restricted thereto.","Examples of the combinations of the fluorescent labels, which are preferred for fluorescent-quenching system utilizing the same mechanism as FRET, include combinations of a dansyl derivative and dabsyl derivative; and a pyrene derivative and a nitrobenzene derivative, but the combinations are not restricted thereto.","The site(s) at which the label(s) is(are) bound is(are) preferably one or both termini of the peptide.","It is preferred to bind different labels, preferably two types of fluorescent labels whose fluorescent wavelengths are different, to the both end regions of the peptide.","It is also preferred to employ a peptide whose one end is cysteine for binding the peptide to the substrate.","Thus, as examples of preferred peptides, those having the structures shown in FIG.","2 are exemplified.","In FIG.","2, A and B denote amino acid residues whose side chains are bound with the fluorescent labels.","In these cases, each of the peptide chains is bound to the substrate through the sulfhydryl group in the side chain of the Cys residue.","Among the peptides represented by FIG.","2, preferred examples are those wherein B is Lys residue whose ε-position is bound with tetramethylrhodamine, and A is Lys residue whose ε-position is bound with fluorescein.","By employing such a structure, the peptide is a labeled peptide having two types of different fluorescent groups at both termini, and measurement utilizing FRET may be attained.","It should be noted that the amino acid residues A and B are not restricted to Lys, but any compounds having functional groups which are relatively highly reactive, such as Orn (ornithine), Glu(glutamic acid) and cysteine may be employed.","By introducing groups for labeling utilizing the highly reactive functional groups, building blocks may be prepared.","In cases where similar reactive functional groups in side chains do not exist in the peptide having the expected spatial structure, the label groups may be introduced site-specifically, after synthesis of the peptide (assembly of amino acid residues).","These synthesis procedures are well-known in the field of peptide synthesis chemistry.","One of the functional groups in each combination shown in Table 1 below may be employed, and so the amino acid is not restricted to Cys.","Thus, if similar reactive functional groups in the side chains do not exist in the peptide having the expected spatial structure, the peptide may be immobilized site-specifically.","The residue such as Cys used for immobilization may be arranged after the C-terminal (after B in the above-mentioned drawing).","That is, among the two structures shown in FIG.","2, the structure shown in the lower location may be employed.","In FIG.","2, the portion indicated as “peptide chain having the expected spatial structure” is the core region mentioned above.","The peptides used in the present invention are those chemically synthesized.","By using a chemically synthesized peptide, the peptide may be synthesized while protecting the functional groups in the side chains of the amino acids, and the synthesized peptide may be bound to the substrate site-specifically.","In cases where the peptide to be immobilized is a protected peptide, it is deprotected after immobilization of the peptide.","It is also possible to immobilize a free (unprotected) peptide.","For example, by arranging Cys residue at the terminal, or by arranging one of avidin and biotin, site-specific binding may be easily attained even if the peptide is unprotected (see Table 1 below).","By this, the peptide may be bound to the substrate at a prescribed site, and the loading amount of the effective peptide (i.e., the peptide which can have the expected spatial structure) may be arbitrarily controlled.","The protected functional groups are usually deprotected after the peptide is bound to the substrate.","Thus, binding of the peptide to the substrate at a site with which the expected spatial structure cannot be attained when it encounters the protein which interacts with the peptide can surely be prevented.","Further, by chemically synthesizing the peptide, the peptide may easily be purified and assayed, so that the loading amount to the substrate may surely be controlled.","Still further, the above-described functional molecule groups or the labels may easily be introduced to the amino acids, and in turn, may easily be bound to the peptide.","Still further, in cases where it is preferred to use an amino acid which does not naturally occur, such an amino acid may easily be incorporated into the peptide.","Thus, the peptides in which all or a part of the amino acid residues constituting the peptide chain are D-amino acids which are not naturally occurring, and the peptides having functional groups other than amino acids, such as heme, flavin and alkyl chains, and the like may also be employed as the peptide to be immobilized, so that the flexibility of measurement is largely increased.","The chemical synthesis per se of the peptide may be carried out by well-known methods, and may easily be carried out by using a commercially available peptide synthesizer.","In this case, it is preferred to carry out the synthesis while protecting the functional groups of the amino acids.","The methods per se for protecting the functional groups in the side chains of amino acids are well-known and described in, for example, Kiyoshi NOKIHARA, Journal of Synthetic Organic Chemistry Japan, 52, 347-358, 1994, “Highly Efficient Peptide Synthesis: Simultaneous and Automatic Synthesis of Multiple Peptides and Peptide Library”; Kiyoshi NOKIHARA and Yoshio YOKOMIZO, Shimadzu Review, 52, 1-8, 1995, “Cleaving Off of Synthetic Peptides by Fmoc-tBu Strategy”; Kiyoshi NOKIHARA, Shimadzu Review, 50, 13-24, 1993, “Reagents for Highly Efficient Solid Phase Synthesis of Peptides”; Kiyoshi NOKIHARA, Shimadzu Review, 50, 3-12, 1993, “Fundamental of Peptide Chemical Synthesis”; Kiyoshi NOKIHARA, Rintaro YAMAMOTO, Hajime HAZAMA, Shin NAKAMURA, Shimadzu Review, 50, 33-43, 1993 “Development of Practical Simultaneous Multiple Solid Phase Peptide Synthesizer PSSM-8”; and K. Nokihara, R. Yamamoto, M. Hazama, O. Wakizawa, S. Nakamura, and M. Yamaguchi, Peptide Chemistry 1991, ed., A. Suzuki, Protein Research Foundation, Osaka, 203-208, 1992, “Development and Applications of a Novel and Practical Simultaneous Multiple Solid Phase Peptide Synthesizer”; Chan, W. C.; White, P. D. Fmoc Solid Phase Peptide Synthesis: A Practical Approach; Oxford University Press: New York, 2000; pp 41-76.The chemically synthesized peptide is then immobilized on the substrate.","This may preferably be attained by reacting the amino group, carboxyl group or the like of the peptide with the amino group, carboxyl group or the like on the substrate thereby binding the peptide to the substrate by covalent bond.","Alternatively, biotin or avidin is bound to the peptide (this may be carried out through the functional group in the side chain of a desired amino acid by a conventional method), and the biotin or avidin may be bound to the avidin or biotin fixed on the substrate.","Examples of the combination of the functional group (or substance) and the functional group (or substance) on the peptide are shown in Table 1.The methods per se for immobilizing the peptide on the substrate through the reaction between the functional groups (or substances) shown in Table 1 are well-known.","The methods for preparing the substrate having the functional groups (or substances) shown in Table 1 are also well-known, and such substrates are commercially available.","The commercially available substrates may preferably be employed.","TABLE 1 Substrate Peptide Reagent for Immobilization Amino group Amino group Glutaraldehyde Amino group Carboxyl group Carbodiimide Carboxyl group Amino group Carbodiimide Sulfhydryl group Bromoacetyl Spontaneously reacts upon mixing.","group Bromoacetyl Sulfhydryl group Spontaneously reacts upon mixing.","group Biotin Avidin Spontaneously reacts upon mixing.","Avidin Biotin Spontaneously reacts upon mixing.","Sulfhydryl group Sulfhydryl group Oxidation (under alkaline pH) Aldehyde group Amino group Spontaneously reacts upon mixing.","Amino group Aldehyde group Spontaneously reacts upon mixing.","Aldehyde group Hydroxylamino Spontaneously reacts upon mixing.","group Hydroxyamino Aldehyde group Spontaneously reacts upon mixing.","group Aldehyde group Hydrazino group Spontaneously reacts upon mixing.","Hydrazino group Aldehyde group Spontaneously reacts upon mixing.","Aldehyde group Sulfhydryl group Spontaneously reacts upon mixing.","Sulfhydryl group Aldehyde group Spontaneously reacts upon mixing.","Active ester Amino group Spontaneously reacts upon mixing.","group Amino group Active ester Spontaneously reacts upon mixing.","group The amount of each peptide to be immobilized on the substrate is not restricted at all, and may be appropriately selected depending on the concentration of the target protein in the test sample, reactivity between the peptide and the target protein and the like.","For example, it is about 1 fmol (1 femto mol) to 1000 nmol, preferably about 0.01 pmol to 1000 pmol.","The binding of the peptide to the substrate may also be carried out by physical adsorption.","In this case, the immobilization of the peptide may be attained by contacting a solution of the peptide in a buffer with the substrate.","In this case, the immobilization reaction may be carried out, for example, at room temperature for about 15 minutes to 2 hours, or at 4° C. overnight, as in the conventional methods.","The concentration of the peptide solution used for immobilization may be appropriately selected depending on the type of the peptide, and type and concentration of the substance to be measured in the test sample.","For example, it is about 1 ng/ml to 100 μg/ml.","After binding the peptide to the substrate, the functional groups of the side chains of the amino acids are deprotected.","The methods for the deprotection are also well-known, and are described in, for example, Kiyoshi NOKIHARA, Journal of Synthetic Organic Chemistry Japan, 52, 347-358, 1994, “Highly Efficient Peptide Synthesis: Simultaneous and Automatic Synthesis of Multiple Peptides and Peptide Library”; Kiyoshi NOKIHARA and Yoshio YOKOMIZO, Shimadzu Review, 52, 1-8, 1995, “Cleaving Off of Synthetic Peptides by Fmoc-tBu Strategy”; Kiyoshi NOKIHARA, Rintaro YAMAMOTO, Hajime HAZAMA, Shin NAKAMURA, Shimadzu Review, 50, 33-43, 1993 “Development of Practical Simultaneous Multiple Solid Phase Peptide Synthesizer PSSM-8”; and K. Nokihara, R. Yamamoto, M. Hazama, O. Wakizawa, S. Nakamura, and M. Yamaguchi, Peptide Chemistry 1991, ed., A. Suzuki, Protein Research Foundation, Osaka, 203-208, 1992, “Development and Applications of a Novel and Practical Simultaneous Multiple Solid Phase Peptide Synthesizer”; Chan, W. C.; White, P. D. Fmoc Solid Phase Peptide Synthesis: A Practical Approach; Oxford University Press: New York, 2000; pp 41-76.In general, the methods for cleavage used in solid phase synthesis (the methods for cleaving peptides from a resin and for simultaneously deprotecting side chains) may be used as they are.","Under the reaction conditions for carrying out the cleavage, the peptide immobilized on the substrate is not cleaved from the substrate.","In the peptide-immobilized substrate according to the present invention, it is preferred to immobilize a plurality of kinds of peptides on a single substrate at specified regions, respectively.","By so doing, a plurality of target proteins may be measured simultaneously.","The number of the kinds of the peptides is not restricted, and usually, it is preferred to immobilize 100 to 10,000 kinds, more preferably about 1000 to 3000 kinds of peptides on a single substrate.","As the substrate, any substrate which can bind the chemically synthesized peptides may be employed without restriction, and glass substrates such as slide glass, may be employed as in the conventional DNA chips.","Substrates having small recesses or grooves may be employed, and wells of microplates may also be used as the substrates.","In cases where the wells of a microplate are used as the substrates, the inner wall of each well constitutes the peptide-immobilized substrate.","In cases where the peptides are immobilized by physical adsorption, it is preferred to use wells of a microplate, or grooves or recesses formed in a chip such as slide glass as the substrate, so that the peptide solution may be poured.","By using a recess as the substrate when the peptides are immobilized by physical adsorption, the measurement may be carried out without problem even if a portion of the peptides are eluted into the solution during the reaction with the target proteins.","The peptide-immobilized substrate according to the present invention may be used for measuring the target protein(s).","The term “measuring” herein includes both detection and quantification.","The measurement of the target protein(s) may be carried out by adding a sample which may contain the target protein(s) onto the peptide-immobilized substrate, allowing the reaction, then washing the chip and measuring the signal.","The reaction with the sample may be carried out, usually, at 4° C. to 40° C. for about 1 minute to 3 hours, preferably at room temperature for about 1 minute to 15 minutes, but the reaction conditions are not restricted thereto.","The sample is not restricted at all.","Examples of the sample include homoginates of animal cells, plant cells, bacterial cells and viruses as well as fractions thereof; body fluids such as blood, serum, plasma, urine, saliva, tissue fluid and spinal fluid; and various foods and beverages, but the samples are not restricted thereto.","By using the peptide-immobilized substrate according to the present invention, the target protein(s) may be measured accurately and simply.","The peptide-immobilized substrate according to the present invention may be used not only for the measurement of the intended target protein(s), but also for the screening of unknown target protein(s).","That is, the number of the types of the basal oligopeptides which can have the particular spatial structures are thought to be about 10.Based on these oligopeptides, when considering the combinations of the sizes, acidities, basicities, aromaticities, aliphaticities and so on, the number of the basal peptides are thought to be about 3000.Thus, after chemically synthesizing these 3000 kinds of peptides and preparing the peptide-immobilized substrate according to the present invention using the peptides, various proteins are reacted with the substrate.","By so doing, what protein reacts with what peptide having what spatial structure may be examined.","By accumulating the results to produce a database, a great number of proteins can be measured, which will largely contribute to the diagnoses of various diseases and developments of new drugs.","Considering the diversity due to incorporation of non-naturally occurring amino acids, the number of the kinds of the oligopeptides exceeds 1,000,000.By virtue of these numerous peptides, the type of recognition may be classified in more detail, so that the accuracy of the diagnosis is thought to be promoted, when the substrate is applied to diagnosis.","By carrying out the method of the present invention on a plurality of chemically synthesized peptides, converting the measured value for each peptide into data with which differences between measured values may be recognized with naked eye, and outputting visible data of the peptides in an array, the protein(s) in the test sample may be measured easily and simply, and the protein(s) contained in an unknown sample may be identified or characterized.","Preferred examples of the data recognized with naked eye include, physical values related to color, such as color, brightness and/or chroma, and it is preferred that the difference in color can be recognized with naked eye depending on the measured value for each peptide.","By this, the measured values can be distinguished with naked eye, and they are outputted in an array.","By so doing, a colored pattern is formed by the measured values for the peptides.","By increasing the number of the chemically synthesized peptides, this color pattern becomes unique to each test sample, so that it serves as if a fingerprint (the method according to the present invention in which the measured values are converted to data which can be recognized with naked eye may be referred to as “fingerprint method” for convenience).","Conversion of the differences among various values into visible data may easily be carried out by using a commercially available computer software.","That is, data processing softwares which enable to make the differences among values be visible are commercially available.","An example thereof is Igor Pro ver.4.04 (commercially available from WaveMetrics, Inc).","By using such a data processing software, for example, by inputting the rate of change of the fluorescence intensities before and after the reaction with the immobilized chemically synthesized peptides as measured values into the software, it is possible to output colors such that, for example, the larger the rate of change, the more yellowish the color (this color can be selected in the software).","Further, the colors corresponding to the measured values are outputted in an array.","Therefore, a colored stripe pattern is formed for each test sample.","Since this colored stripe pattern is unique depending on the type and amount of the protein contained in the test sample, it may be used as a fingerprint which enables to identify or characterize (judging of the characters or properties, identification or estimation of content) the protein contained in an unknown sample.","In this method, a plurality of target proteins may be contained in the test sample.","Even when a plurality of target proteins are contained, since a unique colored pattern is outputted depending on the types and amounts of the target proteins, it can be used as a fingerprint of the sample.","In this case, it is preferred that the chemically synthesized peptides which bind with each of the target proteins be immobilized on the substrate.","However, even if there are no chemically synthesized peptides which bind with one or more of the plurality of target proteins, a colored stripe pattern is formed corresponding thereto, so that lack of the chemically synthesized peptides which bind to one or more target proteins is not necessarily problematic.","In the finger print method, those chemically synthesized peptides which have possibilities to bind with the target protein(s) (i.e., which also have possibilities not to bind with the target protein(s)) may also be immobilized on the substrate.","For example, as described in the Examples below, chemically synthesized peptides each of which has a random region therein may be immobilized.","In such a case, it is unknown what peptide binds with what target protein.","However, anyway, since a unique colored pattern is outputted depending on the type(s) and amount(s) of the target protein(s), it can be used as a fingerprint of the sample.","By carrying out measurements by the fingerprint method using the same substrate on which chemically synthesized peptides are immobilized, accumulating the measured values to produce a database, and by comparing the measured data for an unknown sample with the measured data accumulated in the database, unknown test samples may be identified or characterized.","Since the fingerprint method is also one of the modes of the method according to the present invention using the above-described peptide-immobilized substrate, the above-described explanations about the chemically synthesized peptides, their labels, binding methods, test samples and reaction conditions are applicable to the fingerprint method.","EXAMPLES The present invention will now be described in more detail by way of examples.","It should be noted, however, that the present invention is not restricted to the examples below.","Example 1 As schematically shown in FIG.","3, calmodulin was measured using a peptide chip in which peptides having α-helix structure, each of which both termini were labeled with different fluorescent labels, were immobilized on a substrate.","The procedures were carried out as follows: The sequence of the core region of the peptides, which forms α-helix structure was designed by molecular modeling using a computer (molecular modeling using Insight II/Discover, Molecular Simulation, U.S.) based on the amino acid sequence of the peptide described in a reference (K. T. O'Neil and W. F. DeGrado, Trend Biochem Sci, 15, 59-64 (1990)).","As a result, the amino acid sequence Leu-Lys-Lys-Leu-Leu-Lys-Leu-Leu-Lys-Lys-Leu-Leu-Lys-Leu was employed as the amino acid sequence of the core region.","To this sequence, a Cys residue as an anchor for immobilization, and Glu(Rf1), Lys(Rf2) and Lys(Rf3) as residues labeled with fluorescent labels were added to synthesize Cys-Glu(Rf1)-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Lys(Rf2)-NH2, Glu(Rf1)-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Lys(Rf2)-Cys-NH2, and Lys(Rf3)-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Cys-NH2.Rf1, Rf2 and Rf3 were fluorescent groups, that is, ethyldansyl group, dabsyl group and carboxyfluorescein group, respectively.","Fluorescent amino acid derivatives in which the fluorescent groups (A and B in FIG.","3) were introduced in the side chains of glutamic acid and lysine were synthesized, and used as building blocks.","The carboxylic acid to which a dabsyl group was attached was converted into N-hydroxysuccinimide-activated ester derivative using diisopropylcarbidiimide.","The amino group in the side chain of Fmoc-Lys-OH (Product #04-12-1042, Novabiochem, Switzerland) and the above-described activated ester were stirred overnight at room temperature in dimethylformamide according to a conventional method.","Thereafter, the reaction mixture was concentrated and precipitated with ether to obtain Fmoc-Lys(Rf2)-OH labeled with a fluorescent label (B in FIG.","3).","Yield: 80%.","(Fmoc: fluorenylmethyloxycarbonyl).","Further, fluorescent amino acid Fmoc-Glu(Rf1)-OH obtained by introducing a fluorescent group (A in FIG.","3, ethyldansyl group) into the side chain of the glutamic acid Fmoc-Glu-OBut (tert-butyl ester) (Product #04-12-1075, Novabiochem, Switzerland), was synthesized.","That is, the carboxylic acid in the side chain of Fmoc-Glu-OBut and ethyldansyl were condensed by water-soluble carbodiimide in dioxane-water (3:1), and tert-butyl ester was removed by trifluoroacetic acid, thereby synthesizing Fmoc-glutamic ethyl dansyl.","Yield: 60%.","Further, carboxyfluorescein was converted to N-hydroxysuccinnimide-activated ester derivative using diisopropylcarbidiimide.","The amino group in the side chain of Fmoc-Lys-OH (Product #04-12-1042, Novabiochem, Switzerland) and the above-described activated ester were stirred overnight at room temperature in dimethylformamide according to a conventional method.","Thereafter, the reaction mixture was concentrated and precipitated with ether to obtain Fmoc-Lys(Rf3)-OH labeled with a fluorescent label.","Yield: 85%.","Using these products as the building blocks, the peptides labeled with the labels A and B in the drawing were synthesized in amounts of 15 pmol, respectively, by the solid phase peptide synthesis according to the well-known Fmoc strategy.","That is, using TentaGel SRAM, Product #S30-023 (Rink-amide resin with polyethylene glycol chain) of Rapp Polymere, Germany, as the solid carrier, Fmoc-amino acid derivatives were sequentially condensed.","This was carried out using the method described in Japanese Patent No.","2007165 entitled Multiple Simultaneous Chemical Reaction Apparatus, and using the Multiple Simultaneous Peptide Synthesizer Model PSSM-8 commercially available from Shimadzu Corporation.","The substrate was obtained by bromoacetylation of an aminopropyl glass (Product No.2550, Corning, U.S.).","That is, bromoacetic acid was dissolved in dimethylformamide, and water-soluble carbodiimide (1.05 equivalents) was added thereto under stirring while cooling the mixture in ice.","Twenty minutes later, an aminopropyl glass plate was immersed therein and the mixture was stirred by gently shaking the whole mixture at room temperature for 2 hours.","Then the glass plate was removed and washed with 50% aqueous dimethylformamide solution and tetrahydrofuran, followed by drying in a desiccator under reduced pressrue.","Each of the synthetic peptides (concentration: 0.1 to 10 μM) was immobilized by being reacted with the brom groups on the substrate at room temperature for 30 minutes.","With the system in which the combination of ethyldansyl-dabsyl groups was employed, measurement of fluorescence was performed employing exciting wavelength of 340 nm and detection wavelength of 480 nm.","By adding a calmodulin solution (Tris buffer pH7.4, 150 mM NaCl, 0.1 mM CaCl2, calmodulin concentration: 1.0 μM), about five times increase of the fluorescence intensity was measured using a fluorescence plate reader.","Thus, the protein was able to be detected and quantified.","With the system using the peptide containing carboxylfluorescein, measurement of fluorescence was performed employing exciting wavelength of 490 nm and detection wavelength of 520 nm.","In the presence of the protein, about ten times increase of the fluorescence intensity was measured, so that the protein was able to be detected and quantified.","The calmodulin solution and the peptide chip were reacted at 25° C. for 5 minutes.","Example 2 As schematically shown in FIG.","4, amylase was measured using a peptide chip in which a peptide whose one terminus was labeled with a fluorescent label, which peptide has β-loop structure, was immobilized on a substrate.","The procedures were carried out as follows: As in Example 1, as schematically shown in FIG.","4, a peptide which forms β-loop structure was designed by molecular modeling using a computer based on the amino acid sequence of the peptide described in a reference (F. J. Blanco et al., Eur J Biochem 200, 345-351 (1991), S. Ono et al, Biosci Biotechnol Biochem, 62, 1621-1623 (1998)).","As a result, as the amino acid sequence of the core region, the sequence Tyr-Gln-Ser-Trp-Arg-Tyr-Ser-Gln-Ala was employed.","The peptide having this sequence to which Cys residue as an anchor for immobilization was added to one terminus, and to which Lys residue having a fluorescent group Rf (fluorescent group such as carboxyfluorescein in FIG.","4) was added to another terminus, was synthesized.","Thus, the sequence was Lys(Rf)-Tyr-Gln-Ser-Trp-Arg-Tyr-Ser-Gln-Ala-Cys-NH2.As in Example 1, using the fluorescent amino acid derivative having a fluorescent group (such as carboxyfluorescein) on the side chain of the amino acid as a building block, the peptides were synthesized by the solid phase peptide synthesis method using the Fmoc strategy in an amount of about 15 μmol, respectively.","Each peptide (concentration: 0.1 to 10 μM) in Tris buffer, pH8.0 was immobilized on the plate through thioether bond as in Example 1.When carboxyfluorescein group was used, fluorescence measurement was performed employing exciting wavelength of 490 nm and detection wavelength of 520 nm.","By adding an amylase solution (Tris buffer pH7.4, 150 mM NaCl, amylase concentration: 1.0 μM), about ten times increase of the fluorescence intensity was measured, so that the amylase was able to be detected and quantified.","The amylase solution and the peptide chip were reacted at 25° C. for 5 minutes.","Example 3 Fingerprint Method By the method described in detail below, 126 kinds of peptides having the sequences Ac-Cys-Glu-Thr-Ile-Thr-Val-Xaa-Xaa-Xaa-Xaa-Lys-Thr-Tyr-Lys(F)-Lys-NH2 (wherein “Ac” represents acetyl group and (F) represents a fluorescent label) were synthesized, and each one of the peptides was immobilized on one well in a microplate.","Each of the peptides was reacted with 1) α-amylase, 2) bovine serum albumin (BSA), 3) β-glucosidase, 4) β-galactosidase, 5) lysozyme, 6) cellulase, 7) α-lactoglobulin, 8) defatted almond powder (protein mixture), 9) mixture of 1) to 7) or 10) cell lysate of E. coli and the fluorescence intensities before and after the reaction were measured.","The rate of change of the fluorescence intensities were inputted in Igor Pro ver.4.04 (commercially available from WaveMetrics, Inc), and the colors corresponding to the values (the higher the value, the more yellowish) were outputted in an array.","As a result, colored stripe patterns characteristic to each of the test samples were obtained.","The peptide number was taken along the abscissa such that a lateral width per one peptide was 1.5 mm (therefore, the overall width covering all of the 126 peptides was 1.5×126=189 mm), and the longitudinal length for each peptide was about 5 mm, so that the entire area of the pattern was landscape rectangular.","The region for each peptide (rectangle having a width of 1.5 mm and length of 5 mm) was painted with a color corresponding to the rate of change of the fluorescence intensity, and the regions for each peptide are adjacently arrayed, thereby entirely forming a colored stripe pattern.","The concrete amino acid sequences of the region represented by -Xaa-Xaa-Xaa-Xaa- in the above-described sequence are shown in Table 2 below.","TABLE 2 Peptide Peptide Peptide Peptide No.","Sequence No.","Sequence No.","Sequence No.","Sequence 1 LHGE 33 YHRG 65 YWGP 97 GLRS 2 LPGE 34 RHWG 66 YLHP 98 GWRS 3 RPHE 35 SYWG 67 LRHP 99 REYS 4 YGLE 36 WEYG 68 HGLP 100 RLYS 5 WRLE 37 EHYG 69 LERP 101 GRYS 6 WYLE 38 ERYG 70 GLRP 102 HWYS 7 HGPE 39 LRYG 71 LYRP 103 GLEW 8 LHPE 40 ESYG 72 GESP 104 YHGW 9 HSPE 41 RGEH 73 ELSP 105 RLGW 10 YSPE 42 LYEH 74 ERSP 106 SLGW 11 YWPE 43 WRLH 75 LRSP 107 LEHW 12 LYPE 44 GWLH 76 GLYP 108 YRHW 13 LGSE 45 GRPH 77 SWYP 109 PSHW 14 WHSE 46 RSPH 78 PWER 110 EYHW 15 YHSE 47 LWPH 79 WEGR 111 YGLW 16 PWSE 48 LSRH 80 HLGR 112 EHRW 17 PHWE 49 LWSH 81 YHLR 113 PYRW 18 GPWE 50 LYSH 82 SGPR 114 LESW 19 PGYE 51 GEWH 83 WHPR 115 EYSW 20 RPYE 52 SGYH 84 WSPR 116 HPYW 21 SHEG 53 RYEL 85 WESR 117 RHEY 22 PREG 54 WPGL 86 HWSR 118 HPEY 23 WSEG 55 SGHL 87 SPYR 119 WLHY 24 SPHG 56 HSPL 88 GWYR 120 RPHY 25 WSHG 57 EHRL 89 GRES 121 SPHY 26 YSLG 58 HESL 90 PWGS 122 SRHY 27 WHPG 59 YPSL 91 PYGS 123 WSLY 28 YHPG 60 RWSL 92 RGHS 124 PWLY 29 WRPG 61 PEWL 93 WPLS 125 WERY 30 LSPG 62 PRWL 94 ERLS 126 RSWY 31 RYPG 63 GHYL 95 BYLS 32 LERG 64 RWYL 96 EHRS Detailed Procedures in Example 3 Reagents and Apparatuses All chemicals and solvents were of reagent or HPLC grade, and used without further purification.","Dimethylformamide (DMF) as a solvent for the peptide synthesis was treated with molecular sieve 0.4 nm for overnight.","Fmoc protected amino acid derivatives were purchased from Watanabe Chemical Co., Novabiochem or Shimadzu Scientific Research Inc. Side chain protections were as follows: acetamidomethyl (Acm) or triphenylmethyl (Trt) for Cys; t-butyl (tBu) for Asp, Glu, Thr, Tyr, Ser; Trt for Asn, Gln, His; t-butyloxycarbonyl (Boc) or 4-methyltrityl (Mtt) for Lys; 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl (Pbf) for Arg.","Peptides were synthesized by the solid-phase method by means of Fmoc chemistry.","HPLC was performed on a system composed of a Hitachi D7500 Chromato-Integrator, a L7400 UV-VIS Detector using a Wakosil 5C18 (4.6×150 mm) for analysis or a YMC ODS A323 (10×250 mm) for purification or on a Shimadzu LC2010C system using a Cadenza CD-C18 column (4.6×50 mm) with a linear gradient of acetonitrile/0.1% trifluoroacetic acid (TFA) at a flow rate of 1.0 mL min−1.MALDI-TOFMS was performed on a Shimadzu KOMPACT MALDI III mass spectrometer with 3,5-dimethoxy-4-hydroxycinnamic acid (SA) or α-cyano-4-hydroxycinnamic acid (CHCA) as a matrix.","Amino acid analyses were carried out using a Wakopak WS-PTC column (4.0×200 mm, Wako Pure Chemical Industries) after hydrolysis in 6 M HCl at 110° C. for 24 h in a sealed tube and labeling by phenyl isocyanate (PITC).","Synthesis of Peptides with 15-23 Sequence Assembly of peptide chain: The peptides were assembled on NovaSyn TGR resin (Novabiochem, 0.15 mmol/g).","Fmoc-Tyr(tBu)-Gln(Trt)-Ser(tBu)-Trp-Arg(Pbf)-Tyr(tBu)-Ser(tBu)-Gln(Trt)-Ala-Cys(Acm)-resin (Fmoc(15-23)Acm-resin) and Fmoc-Tyr(tBu)-Gln(Trt)-Ser(tBu)-Trp-Arg(Pbf)-Tyr(tBu)-Ser(tBu)-Gln(Trt)-Ala-Cys(Trt)-resin (Fmoc(15-23)Trt-resin) and Fmoc-Tyr(tBu)-Gln(Trt)-Ser(tBu)-Trp-Arg(Pbf)-Tyr(tBu)-Ser(tBu)-Gln(Trt)-Ala-Lys(Mtt)-resin (Fmoc(15-23)Mtt-resin) were synthesized on an Advanced ChemTech Model 348 MPS peptide synthesizer.","Fmoc-amino acids (6 eq.",")(equivalent may be hereinafter referred to as “eq”.","), 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU, 6 eq.","), HOBt.H2O (6 eq.)","and DIEA (12 eq.)","in NMP were used for coupling of amino acids (for 30 min, twice), and 25% piperidine in NMP was used for Fmoc-removal.","DMF was used as solvent for other procedure such as washing.","Peptides Labeled by a Fluorescein Moiety, F(15-23)Acm and F(15-23)SH: Fmoc(15-23)Acm-resin or Fmoc(15-23)Trt-resin were treated with 20% (v/v) piperidine/NMP to remove a Fmoc group.","Then, 5(6)-carboxyfluorescein (5(6)-FAM, Molecular probes, Inc.) was coupled to N-terminus of peptides using 5(6)-FAM (1.5 eq.","), HBTU (1.5 eq.","), HOBt-H2O (1.5 eq.)","and DIEA (3 eq.)","in DMF for 1.5 h (×2) or 2.5 h (×1).","The resin and the protecting groups except Acm were removed by 1 h treatment of TFA/m-cresol/ethaneditiol/thioanisole (40/1/3/3, v/v) at room temperature.","The peptides were purified by RP-HPLC and characterized by MALDI-TOFMS: F(15-23)Acm m/z 1721.4 ([M+H]+calcd.","1721.8); F(15-23)SH 1649.4 (1649.7).","Peptides Labeled by a Dansyl Moiety, D(15-23)Acm and D(15-23)SH: Fmoc(15-23)Acm-resin or Fmoc(15-23)Trt-resin were treated with 20% (v/v) piperidine/NMP to remove a Fmoc group.","A dansyl moiety was introduced to N-terminus of peptides using 5-dimethylaminonaphthalene-1-sulfonyl chloride (dansyl Cl, 3-6 eq.)","in DMF for 2-3 h. The peptide was cleaved from resin and all protecting groups except Acm were removed according to the procedure same as fluorescein-labeled peptides.","The peptides were purified by RP-HPLC and characterized by MALDI-TOFMS: D(15-23)Acm m/z 1596.3 ([M+H]+ calcd.","1596.8); D(15-23)SH 1525.9 (1525.7).","Peptide Labeled by Fluorescein and EDANS Moieties, F(15-23)ED: Fmoc(15-23)Trt-resin was treated with 20% (v/v) piperidine/NMP to remove a Fmoc group.","A fluorescein moiety was introduced to N-terminus of peptides using 5(6)-carboxyfluorescein succinimisyl ester (5(6)-FAM, SE, Molecular probes, Inc., 1.2 eq.)","and DIEA (2 eq.)","in DMF for 7 h. The peptide was cleaved from resin and all protecting groups were removed according to the procedure same as fluorescein-labeled peptides and purified by RP-HPLC.","The obtained peptide was dissolved in 100 mM Tris-HCl (pH 7.4) and N-(iodoacetaminoethyl)-1-naphthylamine-5′-sulfonic (IAEDANS, Research Organics, 3 eq.)","dissolved in DMF was added.","After 1 h, the reactant was purified by RP-HPLC.","The peptide was characterized by MALDI-TOFMS: F(15-23)ED m/z 1959.2 ([M+H]+ calcd.","1956.0).","Peptides Labeled by a Toumarin Moiety, H(15-23)C or Coumarin and Fluorescein Moieties, F(15-23)C: Fmoc(15-23)Mtt-resin were washed by CHCl3 (four times) and DCM (once) and then treated with DCM/TIS/TFA=94/5/1 (4-6 times for 2 min each) to remove a Mtt group.","After washing the resin with CHCl3 (five times) and NMP (five times), a coumarin moiety was introduced into the side chain amino group of Lys residue using 7-diethylaminocoumarin-3-carboxylilc acid (Fluka, 2 eq.","), HBTU (2 eq.","), HOBt.H2O (2 eq.)","and DIEA (4 eq.)","in NMP for 30 min (×2).","Then, Fmoc group of peptide was removed by the treatment with 20% (v/v) piperidine/NMP and fluorescein moiety was introduced to the N-terminus of peptides by the procedure same as fluorescein-labeled peptides.","The peptide was cleaved from resin and all protecting groups were removed according to the procedure same as described above.","The peptides were purified by RP-HPLC and characterized by MALDI-TOFMS: H(15-23)C m/z 1559.7 ([M+H]+ calcd.","1559.7); F(15-23)C 1919.3 (1918.0).","Synthesis of Peptides with Designed Loop Sequence The peptides were assembled on Rink amide SS resin (Advanced ChemTech, 0.6 mmol/g).","Fmoc-Cys(Trt)-Glu(OtBu)-Thr(tBu)-Ile-Thr(tBu)-Val-Ser(tBu)-Trp-Arg(Pbf)-Tyr(tBu)-Lys(Boc)-Thr(tBu)-Tyr(tBu)-Lys(Mtt)-Lys(Boc)-resin (Fmoc-Loop1 (Mtt)-resin) was synthesized on an Advanced ChemTech Model 348 MPS peptide synthesizer with the same procedure described above.","Each peptide-resins were washed by CHCl3 (four times) and DCM (once) and then treated with DCM/TIS/TFA=94/5/1 (4-6 times for 2 min each) to remove a Mtt group.","After washing the resin with CHCl3 (five times) and DMF (five times), 5(6)-FAM, SE was coupled into the side chain amino group of Lys residue using 5(6)—FAM, SE (3 eq.)","and DIEA.","(1.5 eq.)","overnight.","The peptide was cleaved from resin and all protecting groups were removed according to the procedure as described above.","The peptides were purified by RP-HPLC and characterized by MALDI-TOFMS: Loop1 m/z 2306.9 ([M+H]+calcd.","2306.6).","Fluorescence Spectroscopy Fluorescence spectra were recorded on a Hitachi fluorescence spectrophotometer 850 with a thermo-regulator using a quartz cell with 10 mm pathlength.","All measurements were performed at 25° C., in 20 mM Tris-HCl (pH 7.4) or 20 mM Tris-HCl containing 150 mM NaCl (pH 7.4).","Circular Dichroism Spectroscopy Circular dichroism spectroscopy was performed on a Jasco J-720WI spectropolarimeter with thermo-regulator using a quartz cell with 1 mm pathlength.","All the measurements were performed in 20 mM Tris-HCl containing 150 mM NaCl (pH 7.4) at 25° C. Construction of Peptide Library The designed libraries were synthesized on a module for multiple simultaneous synthesis SRM96 (Shimadzu Scientific Research Inc).","The synthesis was carried out based on the instructions by the manufacturer and the following reference: Reference: Tadashi YASUHARA, Kiyoshi NOKIHARA, Tadasu FURUSHO and Eiko KATAOKA, Tokyo University of Agriculture, Nogaku Shuho (Journal of Tokyo University of Agriculture), 43, 260-267, 1999.The peptide library consisting of 63 library peptides and the peptide with native sequence (SWRY), that is, totally 64 kinds of peptides were synthesized at once.","The peptide chain were assembled on a 4-(2′,4′-dimethoxyphenyl-Fmoc-aminomethyl)phenoxyacetamido-norleusyl-MBHA resin (Rink amide MBHA resin).","The common residues sequence at the C-terminus were coupled assembled in bulk using a Big LibraTube Rt20 (Shimadzu Scientific Research Inc.).","Then, the dried resin was dispensed into the reaction tube of a SRM96 and following residues were coupled on a SRM using HBTU (10 eq.","), HOBt.H2O (10 eq.)","and DIEA (12 eq.).","Protected amino acid (10 eq.)","was reacted as an acyl component.","After assembly of all peptide chain, an Mtt group in the side chain (ε-position) of the protected peptide resin of interest was removed by the treatment with the solution of DCM/TIS/TFA=94/5/1 (for 20 min) and fluorescein moiety were introduced by using 5(6)-FAM (6 eq.)","with HBTU (6 eq.","), HOBt.H2O (6 eq.)","and DIEA (9 eq.","), or 5(6)-FAM (5 eq.)","O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU, 5 eq.)","and DIEA (6 eq.).","The fact that the reaction was completed very specifically was proved by characterization of cleavaged peptides.","Peptide chains were liberated from the resin.","This procedure is called cleavage in this field.","For cleavage of peptides from resin and side chain deprotection, the mixture of TFA/m-cresol/ethaneditiol/thioanisole (40/1/3/3, v/v) was used as a cleavage cocktail.","For 64 peptides, ca.","20 mL of solution was prepared.","To collect the filtrates, a 96-well glass titerplate was equipped under a tube holder.","The cleavage cocktail was added to the resins in reaction tubes dropwise over 30 min, and leave for 1 h. Then resins were washed with small amount of cleavage cocktail.","The filtrates containing peptides were concentrated by N2 blowing, and then diethyl ether was added to precipitate the peptides.","The solution with precipitants were transferred into 1.5 mL polypropylene tube, and centrifuged to collect precipitants.","The precipitant were washed with fresh ether for 5 times and dried in centrifugal evaporator.","The crude peptides were purified by gel permeation chromatography (GPC) using SRM96, of which tubes were filled with Sephadex G-10 (Pharmacia) swelled with 10% (v/v) AcOH aq.","(ca.","100 μL per tube).","Since the peptides labeled with the fluorescent label were colored, the desired fraction was easily obtained by visual observation.","The solution was evaporated to dryness by centrifuging evaporator.","The residual acetic acid was removed by freeze drying with addition of water.","The resulted peptides were dissolved in 200 μL of MeOH and stored with tightly capped at 4° C. Absorption Measurements in 96-Well Plate In order to estimate the concentrations of the above stock solutions of peptides, absorbance at 490 nm of the solutions of prepared peptides were measured by using Benchmark Multiplate Reader (Bio-Rad Laboratories) with 490 rn filter using microtiter plate (Assay Plate, Asahi Techno Glass).","Stock solutions (in MeOH) of peptides were diluted by 20 mM Tris-HCl containing 150 mM NaCl (pH 7.4).","The concentrations of stock solutions were estimated by the comparison of absorbance of each peptide to that of a standard peptide, whose peptide contents was determined by amino acid analysis.","Fluorescence Measurements in 96-Well Plate Fluorescence intensities of the fluorescent-labeled peptides in microtiter plates (Assay Plate, Asahi Techno Glass) as solutions or immobilized to the substrates were measured on a PerSeptive Biosystems CytoFluor™ 4000TR multi-well plate reader at 30° C. Excitation and emission filters used were 485/20 and 530/10 (for fluorescein-labeled peptides), 360/20 and 515/10 (for a dansyl-labeled peptide), 450/50 and 530/10 (for a coumarin and fluorescein labeled peptide), respectively.","Peptide Immobilization to 96-Well Polystyrene Plates Synthesis of Tresyl Activated Dextran (TAD): Dextran (100.0 mg, average MW 64,000-76,000, Sigma D4751) was freeze dried from water (ca.","20 mL).","Dried dextran was dissolved in hexamethylphosphoric triamide (10 mL) at 115° C. in a 50 mL single necked round bottom flask connected with a calcium chloride tube.","The solution was brought to room temperature, and 2,2,2-trifluoroethanesulfonyl chloride (tresyl chloride, 220 μL, 2 mmol, Aldrich 32,478-7) were added dropwise over 1 min under magnetic stirring.","After 15 min, dry pyridine (440 μL, 5.4 mmol) was added dropwise over 1 min.","The product was precipitated from cold ethanol (ca.","20 mL).","The peptide was collected by centrifugation and washed twice with fresh ethanol twice.","Finally the product was dissolved in aqueous acetic acid (1% v/v in water, ca 10 mL), freeze dried and stored at −20° C. Yield; 120.2 mg.","This yield corresponded to 7.4% of the hydroxyl groups in the dextran molecule (one out of 4.5 glucose units) were tresylated and this result was agreeable with the results described in the reference.","Reference: K. Gregorius, S. Mouritsen, H. I. Elsner, J. Immunol.","Meth., 1995, 181, 65-73.Preparation of BrAc Modified Microtiter Plates: A solution of poly-L-lysine (MW 70,000-150,000, Sigma P1274, 0.01 mg/mL) in carbonate buffer (0.1 M, pH 9.6) was added into each well of to polystyrene microtiter plates (Assay Plate, Asahi Techno Glass, 150 μL per well) and the plates were incubated for 2 h at room temperature or overnight at 4° C. The poly-L-lysine coated plates were washed four times with washing buffer (10 mM phosphate, 2.7 mM KCl, 500 mM NaCl, 1% (v/v) Triton X-100, pH 7.2), three times with water, and dried with N2.Then, the above TAD (supra) (0.5 mg/mL) in phosphate buffer (10 mM phosphate, 150 mM NaCl, pH 7.2) was added to the poly-L-lysine coated plates (150 L/well) and incubated for 2 h at 4° C. After washing with water and drying, 1,4-diaminobutane (10 mM, diluted from 1 M stock solution in DMF) in the carbonate buffer was added to the TAD modified plates (100 μL/well) and incubated for 2 h at room temperature.","Following a wash by water, blocking solution (0.1 M 2-aminoethanol in water, pH 8.0 adjusted by HCl) was added (200 μL/well) and incubated for 2 h at room temperature.","The amino-functionalized plates were then washed three times with the washing buffer (supra), three times with water, and dried with N2.Bromoacetic acid was introduced to the amino-functionalized plates using its anhydride.","Bromoacetic acid anhydride was prepared in situ by mixing of BrAcOH (0.6 M) and DIC (0.3 M) in DMF or NMP for 60 min.","It was diluted to 10 mM by water and added to the plate (100 μL/well), and incubated for 5 h at room temperature or overnight at 4° C. The plates were washed three times with water, and further processed as below.","Preparation of Peptide-Bound Microtiter Plates: The peptides with free SH group were dissolved in MeOH as stock solutions (ca.","1 mM).","The peptides solution were diluted to 10 μM by 100 mM Tris buffer (pH 8.0) and immediately added to the bromoacetylated acid-modified plate (100 μL/well).","The plate was incubated for 10 h at room temperature and then washing with the washing buffer for overnight at 4° C. After washing with water and dried with N2, the plate was kept at 4° C. Protein Detection Assay by Using Immobilized Peptides All measurements were performed in 20 mM Tris-HCl containing 150 mM NaCl (pH 7.4).","Purified and partially purified proteins were purchased from Sigma-Aldrich Japan (1.α-amylase from Aspergillus oryzae: A6211, 2.albumin from Bovine serum (BSA): A6793, 3.β-glucosidase from Almonds: G0395, 4.β-galactosidase from Aspergillus oryzae: G5160, 5.lysozyme from Chicken egg white: L6876, 6.cellulase from Aspergillus niger: C1184, 7.β-lactoglobulin from Bovine milk: L3908, 8.almond meal (protein mixture): A3265).","Additionally, cell lysate of of E. coli, was also used.","The solutions of proteins (0.5 mg/mL) were added to the peptide-immobilized plate (100 μL/well), and the plate was incubated at 4° C. for 24 hours.","The fluorescence intensities (1) were measured by using a microplate reader (CytoFluor™) at 30° C. After washing with water, the buffer was added to the plates (100 μL/well) and measured fluorescence intensities (I0) again by using a microplate reader (CytoFluor™) at 30° C. The experiments were repeated twice for each protein and the averages of I/I0 were recorded.","Preparation of E. coli Cell Lysate E. coli.","JA221 (F-, hsdR, DtrpES, leuB6, lacY, recAI) was inoculated onto ca.","5 mL of liquid media composed of 1% trypton, 0.5% yeast extract, and 1% NaCl (w/v) and preincubated at 37° C. for overnight.","Then all of the culture was inoculated into 400 mL of same media and incubated at 37° C. for overnight with vigorous shaking.","The cells were harvested by spinning, and the cell pellets were suspended in the buffer (100 mM potassium phosphate, pH 6.5), followed by washing.","The washed cells were resuspended in 16 mL of the buffer and disrupted by sonication using ultrasonic disintegrator in an ice bath.","Cell debris and larger particles were removed by centrifugation at 12000 rpm for 20 min, and the supernatant was collected.","Total protein concentration of obtained supernatant was estimated as 30 mg/mL, by using Bio-Rad DC Protein Assay kit.","The solution was diluted to a concentration of 0.86 mg/mL with 1.0 mM of phenylmethylsulfonyl fluoride and 5 mM of EDTA, and analyzed by the peptide-immobilized plate.","Processing by Computer of Fingerprint-Like Fluorescence Data The fluorescence data for each protein, obtained by the peptide library immobilized on the plate, were converted into a graph like a fingerprint.","As an example, the case carried out by using a sofware Igor Pro ver.4.04 (WaveMetrics, Inc.) will now be described.","First, the peptides (1-126) were taken along the x-axis.","The line of y=1 was displayed as a line graph for all proteins (10 lines) (1.α-amylase from Aspergillus oryzae, 2.albumin from Bovine serum (BSA), 3.β-glucosidase from Almonds, 4.β-galactosidase from Aspergillus oryzae, 5.lysozyme from Chicken egg white, 6.cellulase from Aspergillus niger, 7.β-lactoglobulin from Bovine milk, 8.almond meal (protein mixture), 9.mixture of 1-7, 10.E.","coli soluble proteins (cell homogenate)).","Then these lines were displayed after shifting the y-axis, and the fluorescence response value (I/I0) for each protein was assigned to each line as the value of z-axis.","As the color gradation, yello-hot was selected from the patterns included in the software.","As a result, characteristic fluorescence response pattern (protein fingerprint) depending on the amino acid sequences of the peptides was obtained for each protein, and the proteins were able to be characterized from the patterns with high reproducibility."]],"string":"[\n [\n \"Peptide-immobilized substrate and method for measuring target protein\",\n \"Disclosed is a peptide-immobilized substrate for measuring a target protein, with which the peptide can have a structure required for being recognized by the target protein, with which the accurate loading amount of the peptide can be attained, and by which a trace amount of the target protein may be measured accurately and simply.\",\n \"The peptide-immobilized substrate for measuring a target protein according to the present invention comprises a chemically synthesized peptide having an.\",\n \"expected spatial structure or having a binding ability with the target protein, which peptide can bind with the target protein and is immobilized on the substrate.\"\n ],\n [\n \"1.A peptide-immobilized substrate for measuring (a) target protein(s), comprising (a) chemically synthesized peptide(s) having (an) expected spatial structure(s) or having (a) binding ability (abilities) with said target protein(s), which peptide(s) can bind with said target protein(s) and which is(are) immobilized on said substrate.\",\n \"2.The peptide-immobilized substrate according to claim 1, wherein said peptide(s) is(are) labeled with (a) label(s) whose signal(s) is(are) changed upon binding of said peptide(s) with said target protein(s).\",\n \"3.The peptide-immobilized substrate according to claim 1, wherein said peptide(s) has(have) (an) amino acid sequence(s) which is(are) so molecularly designed as to attain said expected spatial structure(s).\",\n \"4.The peptide-immobilized substrate according to claim 1, wherein the number of amino acids constituting said peptide is 3 to 50.5.The peptide-immobilized substrate according to claim 2, wherein said label(s) is(are) (a) fluorescent label(s) bound to (an) amino acid(s) located at (a) particular site(s), which amino acid(s) is(are) among the amino acids constituting said peptide.\",\n \"6.The peptide-immobilized substrate according to claim 5, wherein two types of fluorescent labels are bound to particular amino acids located at different sites.\",\n \"7.The peptide-immobilized substrate according to claim 2, wherein said label(s) is(are) bound to one or both terminal regions(s) of said peptide.\",\n \"8.The peptide-immobilized substrate according to claim 1, wherein said peptide(s) is(are) one(those) synthesized while protecting functional groups in the side chains of the amino acids, and is(are) bound to said substrate at (a) desired site(s) by binding the peptide(s) to the substrate while protecting said functional groups which are then deprotected after said peptide(s) is(are) bound to said substrate.\",\n \"9.The peptide-immobilized substrate according to claim 1, wherein the each peptide has (a) functional group(s) at its terminal region, which chemically binds to said substrate, and is bound to said substrate via said functional group.\",\n \"10.The peptide-immobilized substrate according to claim 9, wherein the each peptide has cysteine at one end thereof, and is bound to said substrate through the sulfhydryl group of said cysteine.\",\n \"11.The peptide-immobilized substrate according to claim 1, wherein the each peptide is bound to said substrate by covalent bond.\",\n \"12.The peptide-immobilized substrate according to claim 1, wherein the each peptide has a core region to be subjected to the binding with said target protein, and (a) binding region(s) to be subjected to the binding to said substrate and to the binding with said label(s) in cases where said peptide has said label(s), said core region having spatial structure optimum to the binding with said target protein.\",\n \"13.The peptide-immobilized substrate according to claim 1, wherein a plurality of said peptides are bound to prescribed sites on a single substrate.\",\n \"14.A method for measuring (a) target protein(s) comprising contacting said peptide-immobilized substrate according claim 1 with said target protein(s); and measuring said target protein(s) bound to said peptide(s) based on the change of said signal(s).\",\n \"15.A method for measuring (a) protein(s) in a test sample, comprising the steps of bringing a plurality of chemically synthesized peptides each of which has an expected spatial structure or a binding ability with the target protein, which can or might bind to the target protein, into contact with said test sample which may contain the target protein(s), said plurality of peptides being immobilized on the same or different substrates; measuring the signals which change upon binding between the peptides and the target protein(s); converting each of the values measured for each peptide into data with which differences between measured values may be recognized with naked eye and outputting visible data of the peptides in an array.\",\n \"16.The method according to claim 15, wherein the visible physical value is color, brightness and/or chroma, and differences in color may be recognized with naked eye depending on the measured value for each peptide.\",\n \"17.The method according to claim 15, wherein a plurality of chemically synthesized peptides which bind with a plurality of target proteins are immobilized.\",\n \"18.A method for identifying or characterizing (a) protein(s) in a test sample comprising carrying out the method according to claim 15 for various test samples; accumulating outputted measured values to produce a database; comparing measured data for an unknown test sample with the measured data in said database thereby identifying or characterizing said unknown test sample.\",\n \"19.The method according to claim 15, wherein at least a portion of said plurality of peptides contain random sequences in at least a part of their amino acid sequences.\",\n \"20.The method according to claim 15, wherein at least a portion of said plurality of chemically synthesized peptides are labeled with labels whose signals are changed upon binding of said peptides with said target protein(s).\",\n \"21.The method according to claim 15, wherein at least a portion of said plurality of chemically synthesized peptides have amino acid sequences which are so molecularly designed as to attain said expected spatial structure.\",\n \"22.The method according to claim 15, wherein the number of amino acids constituting each of said plurality of chemically synthesized peptides is 3 to 50.23.The method according to claim 15, wherein each of said labels is a fluorescent label bound to an amino acid located at a particular site, which amino acid is one of the amino acids constituting said peptide.\",\n \"24.The method according to claim 23, wherein two types of fluorescent labels are bound to particular amino acids located at different sites in a single peptide.\",\n \"25.The method according to claim 24, wherein said labels are bound to one or both termini of said single peptide.\",\n \"26.The method according to claim 15, wherein each of said chemically synthesized peptides is one synthesized while protecting functional groups in the side chains of the amino acids, and is bound to said substrate at a desired site by binding the peptide to the substrate while protecting said functional groups which are then deprotected after said peptide is bound to said substrate.\",\n \"27.The method according to claim 15, wherein each of said chemically synthesized peptides has (a) functional group(s) at its terminus, which chemically binds to said substrate, and is bound to said substrate via said functional group.\",\n \"28.The method according to claim 27, wherein each of said chemically synthesized peptides has cysteine at one terminus thereof, and is bound to said substrate through the sulfhydryl group of said cysteine.\",\n \"29.The method according to claim 15, wherein said plurality of chemically synthesized peptides are bound to said substrate by covalent bond.\",\n \"30.The method according to claim 15, wherein each of at least a portion of said plurality of chemically synthesized peptides has a core region to be subjected to the binding with said target protein, and (a) binding region(s) to be subjected to the binding to said substrate and subjected to the binding with said label(s) in cases where said peptide has said label(s), said core region having spatial structure optimum to the binding with said target protein.\"\n ],\n [\n \"<SOH> BACKGROUND ART <EOH>Analyses of functions of proteins existing in the body are important tasks.\",\n \"Methods for identifying and quantifying these expressed proteins satisfy the wide social demands from the clarification of cell functions in the research field to the diagnoses and therapies in the medical field.\",\n \"As means for identifying and quantifying proteins, methods utilizing antibodies and various biosensor systems utilizing arrays of specific proteins are known.\",\n \"The concept for developing the conventional protein chips is based on immobilization of proteins or fragments thereof, whose structures are known.\",\n \"Immobilization of antibody molecules is also based on this concept.\",\n \"By this method, quantification of the loading amount is problematic, and uniform immobilization is difficult.\",\n \"Particularly, since the yield of immobilization (loading amount) is generally low and depends on the properties of the peptide fragments, the method lacks reliability and so wide use thereof is prevented.\",\n \"Further, in cases where protein fragments are used, the sites of cleavage are restricted because of specificities of the cleavage (if the proteins are cleaved without specificities, the resultants are random fragments and so the molecules immobilized on the chips cannot be identified), thus, free design can not be accomplished.\",\n \"Still further, the thus immobilized proteins have structures different from those in the body, and may not have the structures indispensable for attaining the desired recognitions.\",\n \"These are problems in practice.\",\n \"Methods for analyzing interactions between peptides and proteins in solutions using fluorometer or the like are known.\",\n \"However, with these methods, considerable amount of samples are necessary, so that a long time is required for sample preparation.\",\n \"Recently, methods such as surface plasmon or the like are used.\",\n \"However, the apparatuses are expensive, the sensitivities are generally low, and simple measurements at local places cannot be attained.\",\n \"Thus, by the conventional methods, treatments of numerous samples are difficult.\"\n ],\n [\n \"<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG.\",\n \"1 schematically shows the concept of the peptide-immobilized substrate according to the present invention.\",\n \"FIG.\",\n \"2 schematically shows a linear structure of the peptide used in the present invention.\",\n \"FIG.\",\n \"3 schematically shows the measurement carried out in Example 1 of the present invention.\",\n \"FIG.\",\n \"4 schematically shows the measurement carried out in Example 2 of the present invention.\",\n \"detailed-description description=\\\"Detailed Description\\\" end=\\\"lead\\\"?\"\n ],\n [\n \"TECHNICAL FIELD The present invention relates to a peptide-immobilized substrate for measuring (a) target protein(s), and to a method for measuring the target protein(s) using the same.\",\n \"BACKGROUND ART Analyses of functions of proteins existing in the body are important tasks.\",\n \"Methods for identifying and quantifying these expressed proteins satisfy the wide social demands from the clarification of cell functions in the research field to the diagnoses and therapies in the medical field.\",\n \"As means for identifying and quantifying proteins, methods utilizing antibodies and various biosensor systems utilizing arrays of specific proteins are known.\",\n \"The concept for developing the conventional protein chips is based on immobilization of proteins or fragments thereof, whose structures are known.\",\n \"Immobilization of antibody molecules is also based on this concept.\",\n \"By this method, quantification of the loading amount is problematic, and uniform immobilization is difficult.\",\n \"Particularly, since the yield of immobilization (loading amount) is generally low and depends on the properties of the peptide fragments, the method lacks reliability and so wide use thereof is prevented.\",\n \"Further, in cases where protein fragments are used, the sites of cleavage are restricted because of specificities of the cleavage (if the proteins are cleaved without specificities, the resultants are random fragments and so the molecules immobilized on the chips cannot be identified), thus, free design can not be accomplished.\",\n \"Still further, the thus immobilized proteins have structures different from those in the body, and may not have the structures indispensable for attaining the desired recognitions.\",\n \"These are problems in practice.\",\n \"Methods for analyzing interactions between peptides and proteins in solutions using fluorometer or the like are known.\",\n \"However, with these methods, considerable amount of samples are necessary, so that a long time is required for sample preparation.\",\n \"Recently, methods such as surface plasmon or the like are used.\",\n \"However, the apparatuses are expensive, the sensitivities are generally low, and simple measurements at local places cannot be attained.\",\n \"Thus, by the conventional methods, treatments of numerous samples are difficult.\",\n \"DISCLOSURE OF THE INVENTION An object of the present invention is to provide a peptide-immobilized substrate for measuring (a) target protein(s), with which the peptide can have a structure required for being recognized by the target protein, with which the accurate loading amount of the peptide can be attained, and by which (a) trace amount(s) of the target protein(s) may be measured accurately and simply.\",\n \"The present inventors intensively studied to discover that the above-described object may be attained by a peptide-immobilized substrate for measuring a target protein, comprising a chemically synthesized peptide having an expected spatial structures or having a binding ability with the target protein, which the peptide can bind with the target proteins and which is immobilized on the substrate, thereby completing the present invention.\",\n \"Further, the present inventors inferred that a protein in a test sample may be easily and simply measured, or a protein contained in an unknown test sample may be identified or characterized by immobilizing a plurality of chemically synthesized peptides on a substrate; reacting the peptides with a test sample which may contain the target protein; assigning a visible physical value to the signal measured for each of the peptides; and outputting visible data of the peptides in an array.\",\n \"That is, the present invention provides a peptide-immobilized substrate for measuring (a) target protein(s), comprising (a) chemically synthesized peptide(s) having (an) expected spatial structure(s) or having (a) binding ability(abilities) with said target protein(s), which peptide(s) can bind with said target protein(s) and which is(are) immobilized on said substrate.\",\n \"The present invention also provides a method for measuring (a) target protein(s) comprising contacting said peptide-immobilized substrate according to the present invention with said target protein(s); and measuring said target protein(s) bound to said peptide(s) based on the change of said signal(s).\",\n \"The present invention further provides a method for measuring (a) protein(s) in a test sample, comprising the steps of bringing a plurality of chemically synthesized peptides each of which has an expected spatial structure or a binding ability with the target protein, which can or might bind to the target protein, into contact with said test sample which may contain the target protein(s), said plurality of peptides being immobilized on the same or different substrates; measuring the signals which change upon binding between the peptides and the target protein(s); converting each of the values measured for each peptide into data with which differences between measured values may be recognized with naked eye and outputting visible data of the peptides in an array.\",\n \"The present invention still further provides a method for identifying or characterizing (a) protein(s) in a test sample comprising carrying out the method according to the present invention for various test samples; accumulating outputted measured values to produce a database; comparing measured data for an unknown test sample with the measured data in said database thereby identifying or characterizing said unknown test sample.\",\n \"By the present invention, a novel peptide-immobilized substrate for measuring a target protein, by which a trace amount of target protein may be measured accurately and simply, was provided.\",\n \"In the peptide-immobilized substrate according to the present invention, each of the peptides is a chemically synthesized peptide, and may be immobilized on the substrate under the conditions -wherein the functional groups in the side chains of the amino acids are protected or unprotected.\",\n \"That is, the peptide may be immobilized on the substrate at a particular site, and the effective immobilizing amount may be arbitrarily controlled.\",\n \"Further, the peptide may be freely designed, so that an arbitrary label or functional molecular group may be attached to an arbitrary site, or a peptide containing an amino acid which does not occur in natural proteins may be employed.\",\n \"Still further, by the fingerprint method according to the present invention, identification or characterization of the protein contained in the test sample may be attained easily and quickly, and a number of test samples or unknown test samples may be measured.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 schematically shows the concept of the peptide-immobilized substrate according to the present invention.\",\n \"FIG.\",\n \"2 schematically shows a linear structure of the peptide used in the present invention.\",\n \"FIG.\",\n \"3 schematically shows the measurement carried out in Example 1 of the present invention.\",\n \"FIG.\",\n \"4 schematically shows the measurement carried out in Example 2 of the present invention.\",\n \"BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, the peptide to be immobilized on the substrate is a chemically synthesized peptide.\",\n \"Most of the naturally occurring proteins have the properties to molecularly recognize the oligopeptides having particular spatial structures and bind with them (Reference: W. E. Stites, Chem Rev, 97, 1233-1250 (1997), Protein-protein interactions: Interface structure, binding thermodynamics, and mutational analysis; D. P. Fairlie et al, Curr.\",\n \"Medicinal Chem., 5, 29-62 (1998) Toward protein surface mimetics; and A. G. Cochran, Chem & Biol, 7, R85-R94 (2000), Antagonist of protein-protein interactions).\",\n \"It is predetermined what protein binds with what oligopeptide having what spatial structure.\",\n \"The first candidate of the peptide used in the present invention is such a peptide having a particular spatial structure.\",\n \"The spatial structures of the oligopeptide, which bind with proteins include various spatial structures such as α-helix, α-loop-α, β-turn, β-loop-β and β-strand.\",\n \"As the peptide to be immobilized on the substrate, peptides having such a particular spatial structure are exemplified.\",\n \"These spatial structures are those which the peptide may have when the peptide is bound with the target protein, and the peptide does not necessarily have these spatial structures when it is merely immobilized on the substrate.\",\n \"In the present invention, the term “having an expected spatial structure” means that the peptide has the expected spatial structure when bound with the target protein.\",\n \"It should be noted that the entire peptide does not necessarily has the expected spatial structure, as long as most part of the peptide (preferably not less than 60%, more preferably not less than 70% in terms of the number of amino acids) has the expected spatial structure.\",\n \"Therefore, to one or both terminal regions of the peptide, the label(s) hereinbelow described may be bound, and the region(s) may not participate in the formation of the expected spatial structure.\",\n \"It is preferred rather to separate the region(s) (hereinafter also referred to as “binding region”) at which the peptide is immobilized on the substrate or to which the label(s) is(are) attached and the region to be subjected to the binding with the target protein.\",\n \"It is preferred here that the region (hereinafter also referred to as “core region”) to be subjected to the binding with the target protein have the spatial structure which is optimum to the binding.\",\n \"In this case, the binding region(s) is(are) preferably attached to one or both termini of the core region.\",\n \"The peptide having the above-mentioned spatial structure may have a circular structure.\",\n \"In some cases, by making the peptide in a circular form, the binding affinity with the protein may be largely increased.\",\n \"Although a peptide antigen may be chemically synthesized and may be used as the peptide for measuring an antibody, the binding between the peptide and the target protein is not necessarily antigen-antibody reaction at all.\",\n \"As the proteins which bind with the oligopeptides having the above-described spatial structures, the following proteins are exemplified.\",\n \"As the proteins which bind with an oligopeptide having α-helix structure, calmodulin and gp41 of HIV are exemplified.\",\n \"As the proteins which bind with an oligopeptide having β-loop structure, amylase is exemplified.\",\n \"As the proteins which bind with a β-strand peptide, MHC proteins, aspartic acid proteases, HIV-1 protease, cysteine proteases, metalloproteases and serine proteases are exemplified.\",\n \"As the proteins which bind with a β-loop-β or β-turn peptide, fibronectin, LH-RH receptors, substance P receptors, somatostatin receptors and the like are exemplified.\",\n \"But the proteins are not restricted thereto.\",\n \"In addition to the above-described peptides having the expected spatial structures, peptide derivatives having binding abilities with the target proteins, such as those in which an organic molecule group (functional molecule group) known to have an interaction with a protein is attached to a particular site, may also be used as the peptide.\",\n \"Examples of such functional molecule groups include porphyrin derivatives (heme), flavin derivatives, alkyl groups such as retinal and palmityl group, nucleic acid bases and peptide nucleic acids, but the functional molecule groups are not restricted thereto.\",\n \"As the proteins which bind with these functional molecule groups, the following proteins are exemplified.\",\n \"As the proteins which bind with porphyrin derivative (heme)-containing peptides, apo-heme proteins are exemplified.\",\n \"As the proteins which bind with flavin derivative-containing peptides, apo-flavin proteins are exemplified.\",\n \"As the proteins which bind with peptides containing alkyl groups such as retinal and palmityl group, lipocalin proteins are exemplified.\",\n \"As the proteins which bind with the nucleic acid base-containing peptides, DNases, RNases, various gene function-controlling proteins and RNA-binding proteins are exemplified.\",\n \"The concept of the peptide-immobilized substrate comprising a chemically synthesized peptide having an expected spatial structure or having a binding ability with the target protein, which peptide is immobilized on the substrate, is schematically shown in FIG.\",\n \"1.With the technology at present, what amino acid sequence is necessary for attaining a particular spatial structure may be easily determined by using a computer.\",\n \"Computer softwares for carrying out such a molecular designing are commercially available, and the sequence of the peptide having the expected spatial structure may easily be determined by using the computer softwares.\",\n \"The size of the peptide is not restricted, and usually, the number of amino acids is preferably about 2 to 50, more preferably about 5 to 20.The target protein bound with the peptide immobilized on the substrate may be measured by various methods.\",\n \"Examples of these methods include the methods in which the peptide to be immobilized is labeled with (a) label(s) whose signal changes upon binding of the peptide with the target protein, and the target protein is measured based on the change of the label(s); surface plasmon resonance method; Grating method which is a modification of surface plasmon that does not use a prism; and mass-spectrometric analyses, but the methods are not restricted thereto.\",\n \"Among these, the methods in which the peptide to be immobilized is labeled with (a) label(s) whose signal changes upon binding of the peptide with the target protein are preferred because they have the highest sensitivities and reproducibilities.\",\n \"As mentioned above, the peptide used in the present invention is preferably labeled with (a) label(s) which changes the signal upon binding of the peptide with the target protein.\",\n \"Examples of such a label include fluorescent labels, spin labels, UV or visible light-absorbing dyes, radioisotopes, stable isotopes and enzymes which cause chemiluminescent reactions (alkaline phosphatase, acid phosphatase, luciferase, green fluorescence protein and the like), but the labels are not restricted thereto.\",\n \"Among these, fluorescent labels are most preferred.\",\n \"Examples of fluorescent labels include fluorescein and derivatives thereof having the fluorescein structure such as carboxyfluorescein; dansyl group and derivatives thereof having the structure of dansyl group such as ethyldansyl group; coumarin derivatives; pyrene derivatives; and naphthalene derivatives, but the fluorescent labels are not restricted thereto.\",\n \"These fluorescent label reagents per se are known and many of them are commercially available.\",\n \"Further, the methods per se for labeling an amino acid by binding a commercially available fluorescent dye with (a) functional group(s) in the amino acid, such as amino group, are known.\",\n \"For example, the label may easily be introduced by binding the label to the side chain of lysine, glutamic acid or cysteine, or to the amino terminal or carboxyl terminal of the peptide, by the method described in Example 1.Since various fluorescence-labeled building blocks used for the synthesis are commercially available from Molecular Probe, U.S. and so on, these commercially available building blocks may be used as they are.\",\n \"As preferred examples of the fluorescent labels, chemical structures of tetramethylrhodamine and carboxyfluorescein are shown in Formulae [I] and [II] below, respectively.\",\n \"The signals emitted by the above-described labels are changed upon binding of the peptide with the target protein.\",\n \"Although the signal is changed even in cases where a single label is bound to the peptide, it is preferred to bind a plurality of different labels to different sites in the peptide because the change of the signals may be detected more sensitively and so the detection sensitivity may be further promoted.\",\n \"In the especially preferred mode, two types of fluorescent pigments whose fluorescent wavelengths are different are bound to different sites in the peptide.\",\n \"By so doing, sensitive detection may be attained utilizing fluorescence resonance energy transfer (FRET) system.\",\n \"Examples of the combinations of the fluorescent labels, which are preferred for attaining FRET, include combinations of a fluorescein derivative and a rhodamine derivative; a dansyl derivative and a fluorescein derivative; and a pyrene derivative and a fluorescein derivative, but the combinations are not restricted thereto.\",\n \"Examples of the combinations of the fluorescent labels, which are preferred for fluorescent-quenching system utilizing the same mechanism as FRET, include combinations of a dansyl derivative and dabsyl derivative; and a pyrene derivative and a nitrobenzene derivative, but the combinations are not restricted thereto.\",\n \"The site(s) at which the label(s) is(are) bound is(are) preferably one or both termini of the peptide.\",\n \"It is preferred to bind different labels, preferably two types of fluorescent labels whose fluorescent wavelengths are different, to the both end regions of the peptide.\",\n \"It is also preferred to employ a peptide whose one end is cysteine for binding the peptide to the substrate.\",\n \"Thus, as examples of preferred peptides, those having the structures shown in FIG.\",\n \"2 are exemplified.\",\n \"In FIG.\",\n \"2, A and B denote amino acid residues whose side chains are bound with the fluorescent labels.\",\n \"In these cases, each of the peptide chains is bound to the substrate through the sulfhydryl group in the side chain of the Cys residue.\",\n \"Among the peptides represented by FIG.\",\n \"2, preferred examples are those wherein B is Lys residue whose ε-position is bound with tetramethylrhodamine, and A is Lys residue whose ε-position is bound with fluorescein.\",\n \"By employing such a structure, the peptide is a labeled peptide having two types of different fluorescent groups at both termini, and measurement utilizing FRET may be attained.\",\n \"It should be noted that the amino acid residues A and B are not restricted to Lys, but any compounds having functional groups which are relatively highly reactive, such as Orn (ornithine), Glu(glutamic acid) and cysteine may be employed.\",\n \"By introducing groups for labeling utilizing the highly reactive functional groups, building blocks may be prepared.\",\n \"In cases where similar reactive functional groups in side chains do not exist in the peptide having the expected spatial structure, the label groups may be introduced site-specifically, after synthesis of the peptide (assembly of amino acid residues).\",\n \"These synthesis procedures are well-known in the field of peptide synthesis chemistry.\",\n \"One of the functional groups in each combination shown in Table 1 below may be employed, and so the amino acid is not restricted to Cys.\",\n \"Thus, if similar reactive functional groups in the side chains do not exist in the peptide having the expected spatial structure, the peptide may be immobilized site-specifically.\",\n \"The residue such as Cys used for immobilization may be arranged after the C-terminal (after B in the above-mentioned drawing).\",\n \"That is, among the two structures shown in FIG.\",\n \"2, the structure shown in the lower location may be employed.\",\n \"In FIG.\",\n \"2, the portion indicated as “peptide chain having the expected spatial structure” is the core region mentioned above.\",\n \"The peptides used in the present invention are those chemically synthesized.\",\n \"By using a chemically synthesized peptide, the peptide may be synthesized while protecting the functional groups in the side chains of the amino acids, and the synthesized peptide may be bound to the substrate site-specifically.\",\n \"In cases where the peptide to be immobilized is a protected peptide, it is deprotected after immobilization of the peptide.\",\n \"It is also possible to immobilize a free (unprotected) peptide.\",\n \"For example, by arranging Cys residue at the terminal, or by arranging one of avidin and biotin, site-specific binding may be easily attained even if the peptide is unprotected (see Table 1 below).\",\n \"By this, the peptide may be bound to the substrate at a prescribed site, and the loading amount of the effective peptide (i.e., the peptide which can have the expected spatial structure) may be arbitrarily controlled.\",\n \"The protected functional groups are usually deprotected after the peptide is bound to the substrate.\",\n \"Thus, binding of the peptide to the substrate at a site with which the expected spatial structure cannot be attained when it encounters the protein which interacts with the peptide can surely be prevented.\",\n \"Further, by chemically synthesizing the peptide, the peptide may easily be purified and assayed, so that the loading amount to the substrate may surely be controlled.\",\n \"Still further, the above-described functional molecule groups or the labels may easily be introduced to the amino acids, and in turn, may easily be bound to the peptide.\",\n \"Still further, in cases where it is preferred to use an amino acid which does not naturally occur, such an amino acid may easily be incorporated into the peptide.\",\n \"Thus, the peptides in which all or a part of the amino acid residues constituting the peptide chain are D-amino acids which are not naturally occurring, and the peptides having functional groups other than amino acids, such as heme, flavin and alkyl chains, and the like may also be employed as the peptide to be immobilized, so that the flexibility of measurement is largely increased.\",\n \"The chemical synthesis per se of the peptide may be carried out by well-known methods, and may easily be carried out by using a commercially available peptide synthesizer.\",\n \"In this case, it is preferred to carry out the synthesis while protecting the functional groups of the amino acids.\",\n \"The methods per se for protecting the functional groups in the side chains of amino acids are well-known and described in, for example, Kiyoshi NOKIHARA, Journal of Synthetic Organic Chemistry Japan, 52, 347-358, 1994, “Highly Efficient Peptide Synthesis: Simultaneous and Automatic Synthesis of Multiple Peptides and Peptide Library”; Kiyoshi NOKIHARA and Yoshio YOKOMIZO, Shimadzu Review, 52, 1-8, 1995, “Cleaving Off of Synthetic Peptides by Fmoc-tBu Strategy”; Kiyoshi NOKIHARA, Shimadzu Review, 50, 13-24, 1993, “Reagents for Highly Efficient Solid Phase Synthesis of Peptides”; Kiyoshi NOKIHARA, Shimadzu Review, 50, 3-12, 1993, “Fundamental of Peptide Chemical Synthesis”; Kiyoshi NOKIHARA, Rintaro YAMAMOTO, Hajime HAZAMA, Shin NAKAMURA, Shimadzu Review, 50, 33-43, 1993 “Development of Practical Simultaneous Multiple Solid Phase Peptide Synthesizer PSSM-8”; and K. Nokihara, R. Yamamoto, M. Hazama, O. Wakizawa, S. Nakamura, and M. Yamaguchi, Peptide Chemistry 1991, ed., A. Suzuki, Protein Research Foundation, Osaka, 203-208, 1992, “Development and Applications of a Novel and Practical Simultaneous Multiple Solid Phase Peptide Synthesizer”; Chan, W. C.; White, P. D. Fmoc Solid Phase Peptide Synthesis: A Practical Approach; Oxford University Press: New York, 2000; pp 41-76.The chemically synthesized peptide is then immobilized on the substrate.\",\n \"This may preferably be attained by reacting the amino group, carboxyl group or the like of the peptide with the amino group, carboxyl group or the like on the substrate thereby binding the peptide to the substrate by covalent bond.\",\n \"Alternatively, biotin or avidin is bound to the peptide (this may be carried out through the functional group in the side chain of a desired amino acid by a conventional method), and the biotin or avidin may be bound to the avidin or biotin fixed on the substrate.\",\n \"Examples of the combination of the functional group (or substance) and the functional group (or substance) on the peptide are shown in Table 1.The methods per se for immobilizing the peptide on the substrate through the reaction between the functional groups (or substances) shown in Table 1 are well-known.\",\n \"The methods for preparing the substrate having the functional groups (or substances) shown in Table 1 are also well-known, and such substrates are commercially available.\",\n \"The commercially available substrates may preferably be employed.\",\n \"TABLE 1 Substrate Peptide Reagent for Immobilization Amino group Amino group Glutaraldehyde Amino group Carboxyl group Carbodiimide Carboxyl group Amino group Carbodiimide Sulfhydryl group Bromoacetyl Spontaneously reacts upon mixing.\",\n \"group Bromoacetyl Sulfhydryl group Spontaneously reacts upon mixing.\",\n \"group Biotin Avidin Spontaneously reacts upon mixing.\",\n \"Avidin Biotin Spontaneously reacts upon mixing.\",\n \"Sulfhydryl group Sulfhydryl group Oxidation (under alkaline pH) Aldehyde group Amino group Spontaneously reacts upon mixing.\",\n \"Amino group Aldehyde group Spontaneously reacts upon mixing.\",\n \"Aldehyde group Hydroxylamino Spontaneously reacts upon mixing.\",\n \"group Hydroxyamino Aldehyde group Spontaneously reacts upon mixing.\",\n \"group Aldehyde group Hydrazino group Spontaneously reacts upon mixing.\",\n \"Hydrazino group Aldehyde group Spontaneously reacts upon mixing.\",\n \"Aldehyde group Sulfhydryl group Spontaneously reacts upon mixing.\",\n \"Sulfhydryl group Aldehyde group Spontaneously reacts upon mixing.\",\n \"Active ester Amino group Spontaneously reacts upon mixing.\",\n \"group Amino group Active ester Spontaneously reacts upon mixing.\",\n \"group The amount of each peptide to be immobilized on the substrate is not restricted at all, and may be appropriately selected depending on the concentration of the target protein in the test sample, reactivity between the peptide and the target protein and the like.\",\n \"For example, it is about 1 fmol (1 femto mol) to 1000 nmol, preferably about 0.01 pmol to 1000 pmol.\",\n \"The binding of the peptide to the substrate may also be carried out by physical adsorption.\",\n \"In this case, the immobilization of the peptide may be attained by contacting a solution of the peptide in a buffer with the substrate.\",\n \"In this case, the immobilization reaction may be carried out, for example, at room temperature for about 15 minutes to 2 hours, or at 4° C. overnight, as in the conventional methods.\",\n \"The concentration of the peptide solution used for immobilization may be appropriately selected depending on the type of the peptide, and type and concentration of the substance to be measured in the test sample.\",\n \"For example, it is about 1 ng/ml to 100 μg/ml.\",\n \"After binding the peptide to the substrate, the functional groups of the side chains of the amino acids are deprotected.\",\n \"The methods for the deprotection are also well-known, and are described in, for example, Kiyoshi NOKIHARA, Journal of Synthetic Organic Chemistry Japan, 52, 347-358, 1994, “Highly Efficient Peptide Synthesis: Simultaneous and Automatic Synthesis of Multiple Peptides and Peptide Library”; Kiyoshi NOKIHARA and Yoshio YOKOMIZO, Shimadzu Review, 52, 1-8, 1995, “Cleaving Off of Synthetic Peptides by Fmoc-tBu Strategy”; Kiyoshi NOKIHARA, Rintaro YAMAMOTO, Hajime HAZAMA, Shin NAKAMURA, Shimadzu Review, 50, 33-43, 1993 “Development of Practical Simultaneous Multiple Solid Phase Peptide Synthesizer PSSM-8”; and K. Nokihara, R. Yamamoto, M. Hazama, O. Wakizawa, S. Nakamura, and M. Yamaguchi, Peptide Chemistry 1991, ed., A. Suzuki, Protein Research Foundation, Osaka, 203-208, 1992, “Development and Applications of a Novel and Practical Simultaneous Multiple Solid Phase Peptide Synthesizer”; Chan, W. C.; White, P. D. Fmoc Solid Phase Peptide Synthesis: A Practical Approach; Oxford University Press: New York, 2000; pp 41-76.In general, the methods for cleavage used in solid phase synthesis (the methods for cleaving peptides from a resin and for simultaneously deprotecting side chains) may be used as they are.\",\n \"Under the reaction conditions for carrying out the cleavage, the peptide immobilized on the substrate is not cleaved from the substrate.\",\n \"In the peptide-immobilized substrate according to the present invention, it is preferred to immobilize a plurality of kinds of peptides on a single substrate at specified regions, respectively.\",\n \"By so doing, a plurality of target proteins may be measured simultaneously.\",\n \"The number of the kinds of the peptides is not restricted, and usually, it is preferred to immobilize 100 to 10,000 kinds, more preferably about 1000 to 3000 kinds of peptides on a single substrate.\",\n \"As the substrate, any substrate which can bind the chemically synthesized peptides may be employed without restriction, and glass substrates such as slide glass, may be employed as in the conventional DNA chips.\",\n \"Substrates having small recesses or grooves may be employed, and wells of microplates may also be used as the substrates.\",\n \"In cases where the wells of a microplate are used as the substrates, the inner wall of each well constitutes the peptide-immobilized substrate.\",\n \"In cases where the peptides are immobilized by physical adsorption, it is preferred to use wells of a microplate, or grooves or recesses formed in a chip such as slide glass as the substrate, so that the peptide solution may be poured.\",\n \"By using a recess as the substrate when the peptides are immobilized by physical adsorption, the measurement may be carried out without problem even if a portion of the peptides are eluted into the solution during the reaction with the target proteins.\",\n \"The peptide-immobilized substrate according to the present invention may be used for measuring the target protein(s).\",\n \"The term “measuring” herein includes both detection and quantification.\",\n \"The measurement of the target protein(s) may be carried out by adding a sample which may contain the target protein(s) onto the peptide-immobilized substrate, allowing the reaction, then washing the chip and measuring the signal.\",\n \"The reaction with the sample may be carried out, usually, at 4° C. to 40° C. for about 1 minute to 3 hours, preferably at room temperature for about 1 minute to 15 minutes, but the reaction conditions are not restricted thereto.\",\n \"The sample is not restricted at all.\",\n \"Examples of the sample include homoginates of animal cells, plant cells, bacterial cells and viruses as well as fractions thereof; body fluids such as blood, serum, plasma, urine, saliva, tissue fluid and spinal fluid; and various foods and beverages, but the samples are not restricted thereto.\",\n \"By using the peptide-immobilized substrate according to the present invention, the target protein(s) may be measured accurately and simply.\",\n \"The peptide-immobilized substrate according to the present invention may be used not only for the measurement of the intended target protein(s), but also for the screening of unknown target protein(s).\",\n \"That is, the number of the types of the basal oligopeptides which can have the particular spatial structures are thought to be about 10.Based on these oligopeptides, when considering the combinations of the sizes, acidities, basicities, aromaticities, aliphaticities and so on, the number of the basal peptides are thought to be about 3000.Thus, after chemically synthesizing these 3000 kinds of peptides and preparing the peptide-immobilized substrate according to the present invention using the peptides, various proteins are reacted with the substrate.\",\n \"By so doing, what protein reacts with what peptide having what spatial structure may be examined.\",\n \"By accumulating the results to produce a database, a great number of proteins can be measured, which will largely contribute to the diagnoses of various diseases and developments of new drugs.\",\n \"Considering the diversity due to incorporation of non-naturally occurring amino acids, the number of the kinds of the oligopeptides exceeds 1,000,000.By virtue of these numerous peptides, the type of recognition may be classified in more detail, so that the accuracy of the diagnosis is thought to be promoted, when the substrate is applied to diagnosis.\",\n \"By carrying out the method of the present invention on a plurality of chemically synthesized peptides, converting the measured value for each peptide into data with which differences between measured values may be recognized with naked eye, and outputting visible data of the peptides in an array, the protein(s) in the test sample may be measured easily and simply, and the protein(s) contained in an unknown sample may be identified or characterized.\",\n \"Preferred examples of the data recognized with naked eye include, physical values related to color, such as color, brightness and/or chroma, and it is preferred that the difference in color can be recognized with naked eye depending on the measured value for each peptide.\",\n \"By this, the measured values can be distinguished with naked eye, and they are outputted in an array.\",\n \"By so doing, a colored pattern is formed by the measured values for the peptides.\",\n \"By increasing the number of the chemically synthesized peptides, this color pattern becomes unique to each test sample, so that it serves as if a fingerprint (the method according to the present invention in which the measured values are converted to data which can be recognized with naked eye may be referred to as “fingerprint method” for convenience).\",\n \"Conversion of the differences among various values into visible data may easily be carried out by using a commercially available computer software.\",\n \"That is, data processing softwares which enable to make the differences among values be visible are commercially available.\",\n \"An example thereof is Igor Pro ver.4.04 (commercially available from WaveMetrics, Inc).\",\n \"By using such a data processing software, for example, by inputting the rate of change of the fluorescence intensities before and after the reaction with the immobilized chemically synthesized peptides as measured values into the software, it is possible to output colors such that, for example, the larger the rate of change, the more yellowish the color (this color can be selected in the software).\",\n \"Further, the colors corresponding to the measured values are outputted in an array.\",\n \"Therefore, a colored stripe pattern is formed for each test sample.\",\n \"Since this colored stripe pattern is unique depending on the type and amount of the protein contained in the test sample, it may be used as a fingerprint which enables to identify or characterize (judging of the characters or properties, identification or estimation of content) the protein contained in an unknown sample.\",\n \"In this method, a plurality of target proteins may be contained in the test sample.\",\n \"Even when a plurality of target proteins are contained, since a unique colored pattern is outputted depending on the types and amounts of the target proteins, it can be used as a fingerprint of the sample.\",\n \"In this case, it is preferred that the chemically synthesized peptides which bind with each of the target proteins be immobilized on the substrate.\",\n \"However, even if there are no chemically synthesized peptides which bind with one or more of the plurality of target proteins, a colored stripe pattern is formed corresponding thereto, so that lack of the chemically synthesized peptides which bind to one or more target proteins is not necessarily problematic.\",\n \"In the finger print method, those chemically synthesized peptides which have possibilities to bind with the target protein(s) (i.e., which also have possibilities not to bind with the target protein(s)) may also be immobilized on the substrate.\",\n \"For example, as described in the Examples below, chemically synthesized peptides each of which has a random region therein may be immobilized.\",\n \"In such a case, it is unknown what peptide binds with what target protein.\",\n \"However, anyway, since a unique colored pattern is outputted depending on the type(s) and amount(s) of the target protein(s), it can be used as a fingerprint of the sample.\",\n \"By carrying out measurements by the fingerprint method using the same substrate on which chemically synthesized peptides are immobilized, accumulating the measured values to produce a database, and by comparing the measured data for an unknown sample with the measured data accumulated in the database, unknown test samples may be identified or characterized.\",\n \"Since the fingerprint method is also one of the modes of the method according to the present invention using the above-described peptide-immobilized substrate, the above-described explanations about the chemically synthesized peptides, their labels, binding methods, test samples and reaction conditions are applicable to the fingerprint method.\",\n \"EXAMPLES The present invention will now be described in more detail by way of examples.\",\n \"It should be noted, however, that the present invention is not restricted to the examples below.\",\n \"Example 1 As schematically shown in FIG.\",\n \"3, calmodulin was measured using a peptide chip in which peptides having α-helix structure, each of which both termini were labeled with different fluorescent labels, were immobilized on a substrate.\",\n \"The procedures were carried out as follows: The sequence of the core region of the peptides, which forms α-helix structure was designed by molecular modeling using a computer (molecular modeling using Insight II/Discover, Molecular Simulation, U.S.) based on the amino acid sequence of the peptide described in a reference (K. T. O'Neil and W. F. DeGrado, Trend Biochem Sci, 15, 59-64 (1990)).\",\n \"As a result, the amino acid sequence Leu-Lys-Lys-Leu-Leu-Lys-Leu-Leu-Lys-Lys-Leu-Leu-Lys-Leu was employed as the amino acid sequence of the core region.\",\n \"To this sequence, a Cys residue as an anchor for immobilization, and Glu(Rf1), Lys(Rf2) and Lys(Rf3) as residues labeled with fluorescent labels were added to synthesize Cys-Glu(Rf1)-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Lys(Rf2)-NH2, Glu(Rf1)-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Lys(Rf2)-Cys-NH2, and Lys(Rf3)-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Cys-NH2.Rf1, Rf2 and Rf3 were fluorescent groups, that is, ethyldansyl group, dabsyl group and carboxyfluorescein group, respectively.\",\n \"Fluorescent amino acid derivatives in which the fluorescent groups (A and B in FIG.\",\n \"3) were introduced in the side chains of glutamic acid and lysine were synthesized, and used as building blocks.\",\n \"The carboxylic acid to which a dabsyl group was attached was converted into N-hydroxysuccinimide-activated ester derivative using diisopropylcarbidiimide.\",\n \"The amino group in the side chain of Fmoc-Lys-OH (Product #04-12-1042, Novabiochem, Switzerland) and the above-described activated ester were stirred overnight at room temperature in dimethylformamide according to a conventional method.\",\n \"Thereafter, the reaction mixture was concentrated and precipitated with ether to obtain Fmoc-Lys(Rf2)-OH labeled with a fluorescent label (B in FIG.\",\n \"3).\",\n \"Yield: 80%.\",\n \"(Fmoc: fluorenylmethyloxycarbonyl).\",\n \"Further, fluorescent amino acid Fmoc-Glu(Rf1)-OH obtained by introducing a fluorescent group (A in FIG.\",\n \"3, ethyldansyl group) into the side chain of the glutamic acid Fmoc-Glu-OBut (tert-butyl ester) (Product #04-12-1075, Novabiochem, Switzerland), was synthesized.\",\n \"That is, the carboxylic acid in the side chain of Fmoc-Glu-OBut and ethyldansyl were condensed by water-soluble carbodiimide in dioxane-water (3:1), and tert-butyl ester was removed by trifluoroacetic acid, thereby synthesizing Fmoc-glutamic ethyl dansyl.\",\n \"Yield: 60%.\",\n \"Further, carboxyfluorescein was converted to N-hydroxysuccinnimide-activated ester derivative using diisopropylcarbidiimide.\",\n \"The amino group in the side chain of Fmoc-Lys-OH (Product #04-12-1042, Novabiochem, Switzerland) and the above-described activated ester were stirred overnight at room temperature in dimethylformamide according to a conventional method.\",\n \"Thereafter, the reaction mixture was concentrated and precipitated with ether to obtain Fmoc-Lys(Rf3)-OH labeled with a fluorescent label.\",\n \"Yield: 85%.\",\n \"Using these products as the building blocks, the peptides labeled with the labels A and B in the drawing were synthesized in amounts of 15 pmol, respectively, by the solid phase peptide synthesis according to the well-known Fmoc strategy.\",\n \"That is, using TentaGel SRAM, Product #S30-023 (Rink-amide resin with polyethylene glycol chain) of Rapp Polymere, Germany, as the solid carrier, Fmoc-amino acid derivatives were sequentially condensed.\",\n \"This was carried out using the method described in Japanese Patent No.\",\n \"2007165 entitled Multiple Simultaneous Chemical Reaction Apparatus, and using the Multiple Simultaneous Peptide Synthesizer Model PSSM-8 commercially available from Shimadzu Corporation.\",\n \"The substrate was obtained by bromoacetylation of an aminopropyl glass (Product No.2550, Corning, U.S.).\",\n \"That is, bromoacetic acid was dissolved in dimethylformamide, and water-soluble carbodiimide (1.05 equivalents) was added thereto under stirring while cooling the mixture in ice.\",\n \"Twenty minutes later, an aminopropyl glass plate was immersed therein and the mixture was stirred by gently shaking the whole mixture at room temperature for 2 hours.\",\n \"Then the glass plate was removed and washed with 50% aqueous dimethylformamide solution and tetrahydrofuran, followed by drying in a desiccator under reduced pressrue.\",\n \"Each of the synthetic peptides (concentration: 0.1 to 10 μM) was immobilized by being reacted with the brom groups on the substrate at room temperature for 30 minutes.\",\n \"With the system in which the combination of ethyldansyl-dabsyl groups was employed, measurement of fluorescence was performed employing exciting wavelength of 340 nm and detection wavelength of 480 nm.\",\n \"By adding a calmodulin solution (Tris buffer pH7.4, 150 mM NaCl, 0.1 mM CaCl2, calmodulin concentration: 1.0 μM), about five times increase of the fluorescence intensity was measured using a fluorescence plate reader.\",\n \"Thus, the protein was able to be detected and quantified.\",\n \"With the system using the peptide containing carboxylfluorescein, measurement of fluorescence was performed employing exciting wavelength of 490 nm and detection wavelength of 520 nm.\",\n \"In the presence of the protein, about ten times increase of the fluorescence intensity was measured, so that the protein was able to be detected and quantified.\",\n \"The calmodulin solution and the peptide chip were reacted at 25° C. for 5 minutes.\",\n \"Example 2 As schematically shown in FIG.\",\n \"4, amylase was measured using a peptide chip in which a peptide whose one terminus was labeled with a fluorescent label, which peptide has β-loop structure, was immobilized on a substrate.\",\n \"The procedures were carried out as follows: As in Example 1, as schematically shown in FIG.\",\n \"4, a peptide which forms β-loop structure was designed by molecular modeling using a computer based on the amino acid sequence of the peptide described in a reference (F. J. Blanco et al., Eur J Biochem 200, 345-351 (1991), S. Ono et al, Biosci Biotechnol Biochem, 62, 1621-1623 (1998)).\",\n \"As a result, as the amino acid sequence of the core region, the sequence Tyr-Gln-Ser-Trp-Arg-Tyr-Ser-Gln-Ala was employed.\",\n \"The peptide having this sequence to which Cys residue as an anchor for immobilization was added to one terminus, and to which Lys residue having a fluorescent group Rf (fluorescent group such as carboxyfluorescein in FIG.\",\n \"4) was added to another terminus, was synthesized.\",\n \"Thus, the sequence was Lys(Rf)-Tyr-Gln-Ser-Trp-Arg-Tyr-Ser-Gln-Ala-Cys-NH2.As in Example 1, using the fluorescent amino acid derivative having a fluorescent group (such as carboxyfluorescein) on the side chain of the amino acid as a building block, the peptides were synthesized by the solid phase peptide synthesis method using the Fmoc strategy in an amount of about 15 μmol, respectively.\",\n \"Each peptide (concentration: 0.1 to 10 μM) in Tris buffer, pH8.0 was immobilized on the plate through thioether bond as in Example 1.When carboxyfluorescein group was used, fluorescence measurement was performed employing exciting wavelength of 490 nm and detection wavelength of 520 nm.\",\n \"By adding an amylase solution (Tris buffer pH7.4, 150 mM NaCl, amylase concentration: 1.0 μM), about ten times increase of the fluorescence intensity was measured, so that the amylase was able to be detected and quantified.\",\n \"The amylase solution and the peptide chip were reacted at 25° C. for 5 minutes.\",\n \"Example 3 Fingerprint Method By the method described in detail below, 126 kinds of peptides having the sequences Ac-Cys-Glu-Thr-Ile-Thr-Val-Xaa-Xaa-Xaa-Xaa-Lys-Thr-Tyr-Lys(F)-Lys-NH2 (wherein “Ac” represents acetyl group and (F) represents a fluorescent label) were synthesized, and each one of the peptides was immobilized on one well in a microplate.\",\n \"Each of the peptides was reacted with 1) α-amylase, 2) bovine serum albumin (BSA), 3) β-glucosidase, 4) β-galactosidase, 5) lysozyme, 6) cellulase, 7) α-lactoglobulin, 8) defatted almond powder (protein mixture), 9) mixture of 1) to 7) or 10) cell lysate of E. coli and the fluorescence intensities before and after the reaction were measured.\",\n \"The rate of change of the fluorescence intensities were inputted in Igor Pro ver.4.04 (commercially available from WaveMetrics, Inc), and the colors corresponding to the values (the higher the value, the more yellowish) were outputted in an array.\",\n \"As a result, colored stripe patterns characteristic to each of the test samples were obtained.\",\n \"The peptide number was taken along the abscissa such that a lateral width per one peptide was 1.5 mm (therefore, the overall width covering all of the 126 peptides was 1.5×126=189 mm), and the longitudinal length for each peptide was about 5 mm, so that the entire area of the pattern was landscape rectangular.\",\n \"The region for each peptide (rectangle having a width of 1.5 mm and length of 5 mm) was painted with a color corresponding to the rate of change of the fluorescence intensity, and the regions for each peptide are adjacently arrayed, thereby entirely forming a colored stripe pattern.\",\n \"The concrete amino acid sequences of the region represented by -Xaa-Xaa-Xaa-Xaa- in the above-described sequence are shown in Table 2 below.\",\n \"TABLE 2 Peptide Peptide Peptide Peptide No.\",\n \"Sequence No.\",\n \"Sequence No.\",\n \"Sequence No.\",\n \"Sequence 1 LHGE 33 YHRG 65 YWGP 97 GLRS 2 LPGE 34 RHWG 66 YLHP 98 GWRS 3 RPHE 35 SYWG 67 LRHP 99 REYS 4 YGLE 36 WEYG 68 HGLP 100 RLYS 5 WRLE 37 EHYG 69 LERP 101 GRYS 6 WYLE 38 ERYG 70 GLRP 102 HWYS 7 HGPE 39 LRYG 71 LYRP 103 GLEW 8 LHPE 40 ESYG 72 GESP 104 YHGW 9 HSPE 41 RGEH 73 ELSP 105 RLGW 10 YSPE 42 LYEH 74 ERSP 106 SLGW 11 YWPE 43 WRLH 75 LRSP 107 LEHW 12 LYPE 44 GWLH 76 GLYP 108 YRHW 13 LGSE 45 GRPH 77 SWYP 109 PSHW 14 WHSE 46 RSPH 78 PWER 110 EYHW 15 YHSE 47 LWPH 79 WEGR 111 YGLW 16 PWSE 48 LSRH 80 HLGR 112 EHRW 17 PHWE 49 LWSH 81 YHLR 113 PYRW 18 GPWE 50 LYSH 82 SGPR 114 LESW 19 PGYE 51 GEWH 83 WHPR 115 EYSW 20 RPYE 52 SGYH 84 WSPR 116 HPYW 21 SHEG 53 RYEL 85 WESR 117 RHEY 22 PREG 54 WPGL 86 HWSR 118 HPEY 23 WSEG 55 SGHL 87 SPYR 119 WLHY 24 SPHG 56 HSPL 88 GWYR 120 RPHY 25 WSHG 57 EHRL 89 GRES 121 SPHY 26 YSLG 58 HESL 90 PWGS 122 SRHY 27 WHPG 59 YPSL 91 PYGS 123 WSLY 28 YHPG 60 RWSL 92 RGHS 124 PWLY 29 WRPG 61 PEWL 93 WPLS 125 WERY 30 LSPG 62 PRWL 94 ERLS 126 RSWY 31 RYPG 63 GHYL 95 BYLS 32 LERG 64 RWYL 96 EHRS Detailed Procedures in Example 3 Reagents and Apparatuses All chemicals and solvents were of reagent or HPLC grade, and used without further purification.\",\n \"Dimethylformamide (DMF) as a solvent for the peptide synthesis was treated with molecular sieve 0.4 nm for overnight.\",\n \"Fmoc protected amino acid derivatives were purchased from Watanabe Chemical Co., Novabiochem or Shimadzu Scientific Research Inc. Side chain protections were as follows: acetamidomethyl (Acm) or triphenylmethyl (Trt) for Cys; t-butyl (tBu) for Asp, Glu, Thr, Tyr, Ser; Trt for Asn, Gln, His; t-butyloxycarbonyl (Boc) or 4-methyltrityl (Mtt) for Lys; 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl (Pbf) for Arg.\",\n \"Peptides were synthesized by the solid-phase method by means of Fmoc chemistry.\",\n \"HPLC was performed on a system composed of a Hitachi D7500 Chromato-Integrator, a L7400 UV-VIS Detector using a Wakosil 5C18 (4.6×150 mm) for analysis or a YMC ODS A323 (10×250 mm) for purification or on a Shimadzu LC2010C system using a Cadenza CD-C18 column (4.6×50 mm) with a linear gradient of acetonitrile/0.1% trifluoroacetic acid (TFA) at a flow rate of 1.0 mL min−1.MALDI-TOFMS was performed on a Shimadzu KOMPACT MALDI III mass spectrometer with 3,5-dimethoxy-4-hydroxycinnamic acid (SA) or α-cyano-4-hydroxycinnamic acid (CHCA) as a matrix.\",\n \"Amino acid analyses were carried out using a Wakopak WS-PTC column (4.0×200 mm, Wako Pure Chemical Industries) after hydrolysis in 6 M HCl at 110° C. for 24 h in a sealed tube and labeling by phenyl isocyanate (PITC).\",\n \"Synthesis of Peptides with 15-23 Sequence Assembly of peptide chain: The peptides were assembled on NovaSyn TGR resin (Novabiochem, 0.15 mmol/g).\",\n \"Fmoc-Tyr(tBu)-Gln(Trt)-Ser(tBu)-Trp-Arg(Pbf)-Tyr(tBu)-Ser(tBu)-Gln(Trt)-Ala-Cys(Acm)-resin (Fmoc(15-23)Acm-resin) and Fmoc-Tyr(tBu)-Gln(Trt)-Ser(tBu)-Trp-Arg(Pbf)-Tyr(tBu)-Ser(tBu)-Gln(Trt)-Ala-Cys(Trt)-resin (Fmoc(15-23)Trt-resin) and Fmoc-Tyr(tBu)-Gln(Trt)-Ser(tBu)-Trp-Arg(Pbf)-Tyr(tBu)-Ser(tBu)-Gln(Trt)-Ala-Lys(Mtt)-resin (Fmoc(15-23)Mtt-resin) were synthesized on an Advanced ChemTech Model 348 MPS peptide synthesizer.\",\n \"Fmoc-amino acids (6 eq.\",\n \")(equivalent may be hereinafter referred to as “eq”.\",\n \"), 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU, 6 eq.\",\n \"), HOBt.H2O (6 eq.)\",\n \"and DIEA (12 eq.)\",\n \"in NMP were used for coupling of amino acids (for 30 min, twice), and 25% piperidine in NMP was used for Fmoc-removal.\",\n \"DMF was used as solvent for other procedure such as washing.\",\n \"Peptides Labeled by a Fluorescein Moiety, F(15-23)Acm and F(15-23)SH: Fmoc(15-23)Acm-resin or Fmoc(15-23)Trt-resin were treated with 20% (v/v) piperidine/NMP to remove a Fmoc group.\",\n \"Then, 5(6)-carboxyfluorescein (5(6)-FAM, Molecular probes, Inc.) was coupled to N-terminus of peptides using 5(6)-FAM (1.5 eq.\",\n \"), HBTU (1.5 eq.\",\n \"), HOBt-H2O (1.5 eq.)\",\n \"and DIEA (3 eq.)\",\n \"in DMF for 1.5 h (×2) or 2.5 h (×1).\",\n \"The resin and the protecting groups except Acm were removed by 1 h treatment of TFA/m-cresol/ethaneditiol/thioanisole (40/1/3/3, v/v) at room temperature.\",\n \"The peptides were purified by RP-HPLC and characterized by MALDI-TOFMS: F(15-23)Acm m/z 1721.4 ([M+H]+calcd.\",\n \"1721.8); F(15-23)SH 1649.4 (1649.7).\",\n \"Peptides Labeled by a Dansyl Moiety, D(15-23)Acm and D(15-23)SH: Fmoc(15-23)Acm-resin or Fmoc(15-23)Trt-resin were treated with 20% (v/v) piperidine/NMP to remove a Fmoc group.\",\n \"A dansyl moiety was introduced to N-terminus of peptides using 5-dimethylaminonaphthalene-1-sulfonyl chloride (dansyl Cl, 3-6 eq.)\",\n \"in DMF for 2-3 h. The peptide was cleaved from resin and all protecting groups except Acm were removed according to the procedure same as fluorescein-labeled peptides.\",\n \"The peptides were purified by RP-HPLC and characterized by MALDI-TOFMS: D(15-23)Acm m/z 1596.3 ([M+H]+ calcd.\",\n \"1596.8); D(15-23)SH 1525.9 (1525.7).\",\n \"Peptide Labeled by Fluorescein and EDANS Moieties, F(15-23)ED: Fmoc(15-23)Trt-resin was treated with 20% (v/v) piperidine/NMP to remove a Fmoc group.\",\n \"A fluorescein moiety was introduced to N-terminus of peptides using 5(6)-carboxyfluorescein succinimisyl ester (5(6)-FAM, SE, Molecular probes, Inc., 1.2 eq.)\",\n \"and DIEA (2 eq.)\",\n \"in DMF for 7 h. The peptide was cleaved from resin and all protecting groups were removed according to the procedure same as fluorescein-labeled peptides and purified by RP-HPLC.\",\n \"The obtained peptide was dissolved in 100 mM Tris-HCl (pH 7.4) and N-(iodoacetaminoethyl)-1-naphthylamine-5′-sulfonic (IAEDANS, Research Organics, 3 eq.)\",\n \"dissolved in DMF was added.\",\n \"After 1 h, the reactant was purified by RP-HPLC.\",\n \"The peptide was characterized by MALDI-TOFMS: F(15-23)ED m/z 1959.2 ([M+H]+ calcd.\",\n \"1956.0).\",\n \"Peptides Labeled by a Toumarin Moiety, H(15-23)C or Coumarin and Fluorescein Moieties, F(15-23)C: Fmoc(15-23)Mtt-resin were washed by CHCl3 (four times) and DCM (once) and then treated with DCM/TIS/TFA=94/5/1 (4-6 times for 2 min each) to remove a Mtt group.\",\n \"After washing the resin with CHCl3 (five times) and NMP (five times), a coumarin moiety was introduced into the side chain amino group of Lys residue using 7-diethylaminocoumarin-3-carboxylilc acid (Fluka, 2 eq.\",\n \"), HBTU (2 eq.\",\n \"), HOBt.H2O (2 eq.)\",\n \"and DIEA (4 eq.)\",\n \"in NMP for 30 min (×2).\",\n \"Then, Fmoc group of peptide was removed by the treatment with 20% (v/v) piperidine/NMP and fluorescein moiety was introduced to the N-terminus of peptides by the procedure same as fluorescein-labeled peptides.\",\n \"The peptide was cleaved from resin and all protecting groups were removed according to the procedure same as described above.\",\n \"The peptides were purified by RP-HPLC and characterized by MALDI-TOFMS: H(15-23)C m/z 1559.7 ([M+H]+ calcd.\",\n \"1559.7); F(15-23)C 1919.3 (1918.0).\",\n \"Synthesis of Peptides with Designed Loop Sequence The peptides were assembled on Rink amide SS resin (Advanced ChemTech, 0.6 mmol/g).\",\n \"Fmoc-Cys(Trt)-Glu(OtBu)-Thr(tBu)-Ile-Thr(tBu)-Val-Ser(tBu)-Trp-Arg(Pbf)-Tyr(tBu)-Lys(Boc)-Thr(tBu)-Tyr(tBu)-Lys(Mtt)-Lys(Boc)-resin (Fmoc-Loop1 (Mtt)-resin) was synthesized on an Advanced ChemTech Model 348 MPS peptide synthesizer with the same procedure described above.\",\n \"Each peptide-resins were washed by CHCl3 (four times) and DCM (once) and then treated with DCM/TIS/TFA=94/5/1 (4-6 times for 2 min each) to remove a Mtt group.\",\n \"After washing the resin with CHCl3 (five times) and DMF (five times), 5(6)-FAM, SE was coupled into the side chain amino group of Lys residue using 5(6)—FAM, SE (3 eq.)\",\n \"and DIEA.\",\n \"(1.5 eq.)\",\n \"overnight.\",\n \"The peptide was cleaved from resin and all protecting groups were removed according to the procedure as described above.\",\n \"The peptides were purified by RP-HPLC and characterized by MALDI-TOFMS: Loop1 m/z 2306.9 ([M+H]+calcd.\",\n \"2306.6).\",\n \"Fluorescence Spectroscopy Fluorescence spectra were recorded on a Hitachi fluorescence spectrophotometer 850 with a thermo-regulator using a quartz cell with 10 mm pathlength.\",\n \"All measurements were performed at 25° C., in 20 mM Tris-HCl (pH 7.4) or 20 mM Tris-HCl containing 150 mM NaCl (pH 7.4).\",\n \"Circular Dichroism Spectroscopy Circular dichroism spectroscopy was performed on a Jasco J-720WI spectropolarimeter with thermo-regulator using a quartz cell with 1 mm pathlength.\",\n \"All the measurements were performed in 20 mM Tris-HCl containing 150 mM NaCl (pH 7.4) at 25° C. Construction of Peptide Library The designed libraries were synthesized on a module for multiple simultaneous synthesis SRM96 (Shimadzu Scientific Research Inc).\",\n \"The synthesis was carried out based on the instructions by the manufacturer and the following reference: Reference: Tadashi YASUHARA, Kiyoshi NOKIHARA, Tadasu FURUSHO and Eiko KATAOKA, Tokyo University of Agriculture, Nogaku Shuho (Journal of Tokyo University of Agriculture), 43, 260-267, 1999.The peptide library consisting of 63 library peptides and the peptide with native sequence (SWRY), that is, totally 64 kinds of peptides were synthesized at once.\",\n \"The peptide chain were assembled on a 4-(2′,4′-dimethoxyphenyl-Fmoc-aminomethyl)phenoxyacetamido-norleusyl-MBHA resin (Rink amide MBHA resin).\",\n \"The common residues sequence at the C-terminus were coupled assembled in bulk using a Big LibraTube Rt20 (Shimadzu Scientific Research Inc.).\",\n \"Then, the dried resin was dispensed into the reaction tube of a SRM96 and following residues were coupled on a SRM using HBTU (10 eq.\",\n \"), HOBt.H2O (10 eq.)\",\n \"and DIEA (12 eq.).\",\n \"Protected amino acid (10 eq.)\",\n \"was reacted as an acyl component.\",\n \"After assembly of all peptide chain, an Mtt group in the side chain (ε-position) of the protected peptide resin of interest was removed by the treatment with the solution of DCM/TIS/TFA=94/5/1 (for 20 min) and fluorescein moiety were introduced by using 5(6)-FAM (6 eq.)\",\n \"with HBTU (6 eq.\",\n \"), HOBt.H2O (6 eq.)\",\n \"and DIEA (9 eq.\",\n \"), or 5(6)-FAM (5 eq.)\",\n \"O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU, 5 eq.)\",\n \"and DIEA (6 eq.).\",\n \"The fact that the reaction was completed very specifically was proved by characterization of cleavaged peptides.\",\n \"Peptide chains were liberated from the resin.\",\n \"This procedure is called cleavage in this field.\",\n \"For cleavage of peptides from resin and side chain deprotection, the mixture of TFA/m-cresol/ethaneditiol/thioanisole (40/1/3/3, v/v) was used as a cleavage cocktail.\",\n \"For 64 peptides, ca.\",\n \"20 mL of solution was prepared.\",\n \"To collect the filtrates, a 96-well glass titerplate was equipped under a tube holder.\",\n \"The cleavage cocktail was added to the resins in reaction tubes dropwise over 30 min, and leave for 1 h. Then resins were washed with small amount of cleavage cocktail.\",\n \"The filtrates containing peptides were concentrated by N2 blowing, and then diethyl ether was added to precipitate the peptides.\",\n \"The solution with precipitants were transferred into 1.5 mL polypropylene tube, and centrifuged to collect precipitants.\",\n \"The precipitant were washed with fresh ether for 5 times and dried in centrifugal evaporator.\",\n \"The crude peptides were purified by gel permeation chromatography (GPC) using SRM96, of which tubes were filled with Sephadex G-10 (Pharmacia) swelled with 10% (v/v) AcOH aq.\",\n \"(ca.\",\n \"100 μL per tube).\",\n \"Since the peptides labeled with the fluorescent label were colored, the desired fraction was easily obtained by visual observation.\",\n \"The solution was evaporated to dryness by centrifuging evaporator.\",\n \"The residual acetic acid was removed by freeze drying with addition of water.\",\n \"The resulted peptides were dissolved in 200 μL of MeOH and stored with tightly capped at 4° C. Absorption Measurements in 96-Well Plate In order to estimate the concentrations of the above stock solutions of peptides, absorbance at 490 nm of the solutions of prepared peptides were measured by using Benchmark Multiplate Reader (Bio-Rad Laboratories) with 490 rn filter using microtiter plate (Assay Plate, Asahi Techno Glass).\",\n \"Stock solutions (in MeOH) of peptides were diluted by 20 mM Tris-HCl containing 150 mM NaCl (pH 7.4).\",\n \"The concentrations of stock solutions were estimated by the comparison of absorbance of each peptide to that of a standard peptide, whose peptide contents was determined by amino acid analysis.\",\n \"Fluorescence Measurements in 96-Well Plate Fluorescence intensities of the fluorescent-labeled peptides in microtiter plates (Assay Plate, Asahi Techno Glass) as solutions or immobilized to the substrates were measured on a PerSeptive Biosystems CytoFluor™ 4000TR multi-well plate reader at 30° C. Excitation and emission filters used were 485/20 and 530/10 (for fluorescein-labeled peptides), 360/20 and 515/10 (for a dansyl-labeled peptide), 450/50 and 530/10 (for a coumarin and fluorescein labeled peptide), respectively.\",\n \"Peptide Immobilization to 96-Well Polystyrene Plates Synthesis of Tresyl Activated Dextran (TAD): Dextran (100.0 mg, average MW 64,000-76,000, Sigma D4751) was freeze dried from water (ca.\",\n \"20 mL).\",\n \"Dried dextran was dissolved in hexamethylphosphoric triamide (10 mL) at 115° C. in a 50 mL single necked round bottom flask connected with a calcium chloride tube.\",\n \"The solution was brought to room temperature, and 2,2,2-trifluoroethanesulfonyl chloride (tresyl chloride, 220 μL, 2 mmol, Aldrich 32,478-7) were added dropwise over 1 min under magnetic stirring.\",\n \"After 15 min, dry pyridine (440 μL, 5.4 mmol) was added dropwise over 1 min.\",\n \"The product was precipitated from cold ethanol (ca.\",\n \"20 mL).\",\n \"The peptide was collected by centrifugation and washed twice with fresh ethanol twice.\",\n \"Finally the product was dissolved in aqueous acetic acid (1% v/v in water, ca 10 mL), freeze dried and stored at −20° C. Yield; 120.2 mg.\",\n \"This yield corresponded to 7.4% of the hydroxyl groups in the dextran molecule (one out of 4.5 glucose units) were tresylated and this result was agreeable with the results described in the reference.\",\n \"Reference: K. Gregorius, S. Mouritsen, H. I. Elsner, J. Immunol.\",\n \"Meth., 1995, 181, 65-73.Preparation of BrAc Modified Microtiter Plates: A solution of poly-L-lysine (MW 70,000-150,000, Sigma P1274, 0.01 mg/mL) in carbonate buffer (0.1 M, pH 9.6) was added into each well of to polystyrene microtiter plates (Assay Plate, Asahi Techno Glass, 150 μL per well) and the plates were incubated for 2 h at room temperature or overnight at 4° C. The poly-L-lysine coated plates were washed four times with washing buffer (10 mM phosphate, 2.7 mM KCl, 500 mM NaCl, 1% (v/v) Triton X-100, pH 7.2), three times with water, and dried with N2.Then, the above TAD (supra) (0.5 mg/mL) in phosphate buffer (10 mM phosphate, 150 mM NaCl, pH 7.2) was added to the poly-L-lysine coated plates (150 L/well) and incubated for 2 h at 4° C. After washing with water and drying, 1,4-diaminobutane (10 mM, diluted from 1 M stock solution in DMF) in the carbonate buffer was added to the TAD modified plates (100 μL/well) and incubated for 2 h at room temperature.\",\n \"Following a wash by water, blocking solution (0.1 M 2-aminoethanol in water, pH 8.0 adjusted by HCl) was added (200 μL/well) and incubated for 2 h at room temperature.\",\n \"The amino-functionalized plates were then washed three times with the washing buffer (supra), three times with water, and dried with N2.Bromoacetic acid was introduced to the amino-functionalized plates using its anhydride.\",\n \"Bromoacetic acid anhydride was prepared in situ by mixing of BrAcOH (0.6 M) and DIC (0.3 M) in DMF or NMP for 60 min.\",\n \"It was diluted to 10 mM by water and added to the plate (100 μL/well), and incubated for 5 h at room temperature or overnight at 4° C. The plates were washed three times with water, and further processed as below.\",\n \"Preparation of Peptide-Bound Microtiter Plates: The peptides with free SH group were dissolved in MeOH as stock solutions (ca.\",\n \"1 mM).\",\n \"The peptides solution were diluted to 10 μM by 100 mM Tris buffer (pH 8.0) and immediately added to the bromoacetylated acid-modified plate (100 μL/well).\",\n \"The plate was incubated for 10 h at room temperature and then washing with the washing buffer for overnight at 4° C. After washing with water and dried with N2, the plate was kept at 4° C. Protein Detection Assay by Using Immobilized Peptides All measurements were performed in 20 mM Tris-HCl containing 150 mM NaCl (pH 7.4).\",\n \"Purified and partially purified proteins were purchased from Sigma-Aldrich Japan (1.α-amylase from Aspergillus oryzae: A6211, 2.albumin from Bovine serum (BSA): A6793, 3.β-glucosidase from Almonds: G0395, 4.β-galactosidase from Aspergillus oryzae: G5160, 5.lysozyme from Chicken egg white: L6876, 6.cellulase from Aspergillus niger: C1184, 7.β-lactoglobulin from Bovine milk: L3908, 8.almond meal (protein mixture): A3265).\",\n \"Additionally, cell lysate of of E. coli, was also used.\",\n \"The solutions of proteins (0.5 mg/mL) were added to the peptide-immobilized plate (100 μL/well), and the plate was incubated at 4° C. for 24 hours.\",\n \"The fluorescence intensities (1) were measured by using a microplate reader (CytoFluor™) at 30° C. After washing with water, the buffer was added to the plates (100 μL/well) and measured fluorescence intensities (I0) again by using a microplate reader (CytoFluor™) at 30° C. The experiments were repeated twice for each protein and the averages of I/I0 were recorded.\",\n \"Preparation of E. coli Cell Lysate E. coli.\",\n \"JA221 (F-, hsdR, DtrpES, leuB6, lacY, recAI) was inoculated onto ca.\",\n \"5 mL of liquid media composed of 1% trypton, 0.5% yeast extract, and 1% NaCl (w/v) and preincubated at 37° C. for overnight.\",\n \"Then all of the culture was inoculated into 400 mL of same media and incubated at 37° C. for overnight with vigorous shaking.\",\n \"The cells were harvested by spinning, and the cell pellets were suspended in the buffer (100 mM potassium phosphate, pH 6.5), followed by washing.\",\n \"The washed cells were resuspended in 16 mL of the buffer and disrupted by sonication using ultrasonic disintegrator in an ice bath.\",\n \"Cell debris and larger particles were removed by centrifugation at 12000 rpm for 20 min, and the supernatant was collected.\",\n \"Total protein concentration of obtained supernatant was estimated as 30 mg/mL, by using Bio-Rad DC Protein Assay kit.\",\n \"The solution was diluted to a concentration of 0.86 mg/mL with 1.0 mM of phenylmethylsulfonyl fluoride and 5 mM of EDTA, and analyzed by the peptide-immobilized plate.\",\n \"Processing by Computer of Fingerprint-Like Fluorescence Data The fluorescence data for each protein, obtained by the peptide library immobilized on the plate, were converted into a graph like a fingerprint.\",\n \"As an example, the case carried out by using a sofware Igor Pro ver.4.04 (WaveMetrics, Inc.) will now be described.\",\n \"First, the peptides (1-126) were taken along the x-axis.\",\n \"The line of y=1 was displayed as a line graph for all proteins (10 lines) (1.α-amylase from Aspergillus oryzae, 2.albumin from Bovine serum (BSA), 3.β-glucosidase from Almonds, 4.β-galactosidase from Aspergillus oryzae, 5.lysozyme from Chicken egg white, 6.cellulase from Aspergillus niger, 7.β-lactoglobulin from Bovine milk, 8.almond meal (protein mixture), 9.mixture of 1-7, 10.E.\",\n \"coli soluble proteins (cell homogenate)).\",\n \"Then these lines were displayed after shifting the y-axis, and the fluorescence response value (I/I0) for each protein was assigned to each line as the value of z-axis.\",\n \"As the color gradation, yello-hot was selected from the patterns included in the software.\",\n \"As a result, characteristic fluorescence response pattern (protein fingerprint) depending on the amino acid sequences of the peptides was obtained for each protein, and the proteins were able to be characterized from the patterns with high reproducibility.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476861"}}},{"rowIdx":978,"cells":{"text":{"kind":"list like","value":[["Method and apparatus for parameter estimation, modulation classification and interference characterization in satellite communication systems","A digital signal processing (DSP)-based approach to parameter estimation modulation identification and interference charaterization in connection with a satellite Communication Monitoring System (CSM).","The techniques descried here also allow automatic generation of satellite frequency plans (S74) without any a priori knowledge of such plans.","Individual processes for carrier isolation (S71), segmentation (S72), frequency estimation (S73), symbol rate estimation, bit error rate estimation, modulation identification and interference characterization are disclosed may be combined in a totally automated process."],["1.A method of automatically isolating carriers of a composite waveform having a bandwidth in a satellite communication system, comprising: a) performing an FFT processing of size NFFT on the composite waveform, where the value of NFFT is programmable; b) obtaining a power spectrum from the FFT processing by computing the squared magnitude of each FFT coefficient; c) repeating step b) a plurality of times and averaging the results; d) setting, a noise floor pn; e) filtering the power spectrum; f) setting a minimum carrier level pc X dB above the noise floor pn, where X is a programmable parameter, g) identifying the lower and upper frequency limits for each carrier; and h) digitally filtering the individual carriers.","2.The method of claim 1 wherein, when a value higher than pc is first detected at a certain frequency, that frequency is taken as the lower frequency limit of a carrier and when the value at the filter output drops first to a level below pc, the corresponding frequency is taken as the upper frequency limit of the carrier.","3.The method of claim 1 wherein.","The processing proceeds through individual frequency points within the spectrum.","4.A method of automatically providing segmentation of time domain data record in a satellite communication system comprising: a) determining a number of samples that are to be included in a segment; b) computing, the instantaneous power in each sample; c) filtering instantaneous power values; d) computing an average and a standard deviation of the instantaneous power; e) computing a normalized standard deviation and comparing the normalized standard deviation to a threshold; f) determining whether the threshold is exceeded and if not exceeded, the segment is rejected, otherwise, it is accepted; and g) repeating the foregoing process for at least one additional segment.","5.The method of claim 4 wherein the determining step includes identifying a suitable power of 2 to represent the selected number of samples in the segment.","6.The method of claim 4 wherein the threshold of step e) is based on a value of Eb/No, where Eb is the signal strength and No is a corresponding noise value.","7.A method of automatically estimating frequency in a satellite communication system comprising: a) computing a frequency estimate; b) providing a modified an instantaneous phase method; c) computing a second frequency estimate is computed using the modified instantaneous phase method; d) detecting any harmonics in the spectrum; e) enhancing a FFT-based location of harmonics by using unbiased interpolation of the FFT coefficients; f) computing a third frequency estimate on the basis of the enhanced harmonics location process; g) determining a weighted average of the frequency estimates; h) assigning weights on the basis of spectral symmetry, envelope fluctuations, and strength of the frequency harmonics.","8.The method of claim 7 wherein step a) uses the centroid method.","9.The method of claim 7 wherein the modifying step comprises assigning a weight proportional to the square of the magnitude to each instantaneous phase value, so that samples with higher SNR are given more weight.","10.The method of claim 7 wherein the waveform is passed through a non-linearity 11.The method of claim 7 further comprising using a phase locked loop (PLL) to track the received carrier.","12.A method of automatically estimating symbol rate in a satellite communication system comprising: a) applying a delay and multiply technique wherein both a received signal and its delayed replica are passed through a non-linearity to produce harmonics at the symbol rate; b) computing the number of crossings per unit time where a signal envelope crosses a half power level; c) tracking the timing of the received waveform; d) providing non-uniform sampling at a non-uniform sampling rate that is slowly and monotonically increasing, and covers the range of uncertainty in the symbol rate; e) once lock is achieved, resuming uniform sampling; and f) using a PLL to fine tune the symbol rate estimate.","13.The method of claim 12 wherein the timing step comprises tracking clock frequency and phase, using a second order PLL.","14.The method of claim 12 wherein said non-uniform sampling comprises generating a set by digital interpolation between the available uniformly sampled samples.","15.A method of automatically estimating bit error rate on received signals in a satellite communication system, comprising: a) processing the received signals with a properly matched and equalized filter; b) tracking of the carrier phase and the clock phase; c) using maximum likelihood techniques to estimate one or more of the phase noise, intermodulation products, quadrature imbalances, and non-linearity's.","d) constructing a waveform with estimated parameters and modulation type; e) subjecting the waveform to estimated impairments, f) estimating the bit error rate.","16.The method of claim 15 further comprising, estimating the waveform parameters and determining the modulation type.","17.The method of claim 15 further comprising automatically considering side information regarding the transmitter characteristics.","18.The method of claim 15 further comprising constructing a noise-free scattering diagram based on the estimated impairments, estimating the uncoded and coded BER, using maximum likelihood, from the noise-free scattering diagram, and obtaining an estimated Eb/No, where Eb is the signal strength and No is a corresponding noise value.","19.A method of automatically classifying modulation of a received signal in a satellite communication system, comprising: a) estimating the parameters of the received signal waveform; b) estimating the signal-to-noise ratio (SNR) of the received signal waveform; c) assigning each sample contributing to a key feature a weight proportional to its SNR; d) modifying the key features computation such that each sample contributing to a key feature is assigned a weight proportional to its distance from the symbol edges; e) modifying a sub-optimum hierarchical classification approach to a vector approach, wherein several features are applied simultaneously to a multidimensional threshold; f) making the number of segments processed SNR-dependent; g) for each segment processed, assign a ranking as to how likely it is that the waveform under examination belongs to each of the modulation classes under consideration; and h) soft combining all the segment rankings to arrive at the most likely overall classification of a modulation type.","20.The method of claim 19, further comprising using side information to narrow down the set of potential modulation formats.","21.The method of claim 19 wherein, in the modifying step e) the threshold setting is made SNR-dependent; 22.The method of claim 19 wherein the side information is input automatically from a data base.","23.A method of automatically characterizing interference in a satellite communication system comprising: a) obtaining waveform parameters and modulation type of the desired signal, b) processing the received samples with a properly matched and equalized filter; c) tracking the carrier phase and the clock phase; d) estimating at least one of the phase noise, intermodulation products, quadrature imbalances, and non-linearity's using maximum likelihood techniques; e) demodulating the received signal and recovering the transmitted bits; f) remodulating the transmitted bits on a carrier according to the modulation type, symbol rate, and filter characteristics; and g) performing a correlation and spectral analysis on the residual signal to extract interferer information from the noise.","24.The method of claim 23 further comprising estimating the parameters.","25.The method of claim 23 further comprising and automatically determining modulation type.","26.The method of claim 23 further comprising using side information available regarding the transmitter characteristics.","27.The method of claim 23 wherein, if the SNR is low and the error rate is high, applying FEC decoding of the signal to recover the information bits; and re-encoding the information bits.","28.A method of automatically generating a satellite frequency plan in a satellite system based on signals received from a satellite, comprising: a) isolating the carrier is isolated automatically; b) segmentation processing the received signal; and c) automatically estimating the frequency d) automatically combining the result of carrier isolation, segmentation and frequency estimation to develop a frequency plan for the satellite.","29.An automated CSM system for use in a satellite communication system and operative to implement any one of the methods set forth in claim 1."],["<SOH> BACKGROUND <EOH>In a satellite communication system, particularly a system where the satellite is deployed in a geostationary orbit, the satellite will be able to receive signals transmitted to the satellite by earth stations at an allocated uplink frequency band and will be operative to transmit signals to earth stations on allocated downlink frequency bands.","The uplink bands are selected to be spaced apart from the downlink bands in order to avoid interference.","Nonetheless, interference may be generated due to transmissions from adjacent earth stations or adjacent satellites having overlapping beams.","In addition, interference may arise from natural phenomena, such as rainfall, scattering, terrestrial communications and the like.","With respect to the downlink, the signal received at an earth station from the satellite is frequency-down-converted and digitized by means of an analog-to-digital (A/D) converter.","A typical value for the bandwidth of the A/D converted frequency band is 36 MHz.","The A/D converter output is a stream of bits which essentially captures all the information in the received signal.","FIG.","1 illustrates a satellite system 10 having a plurality of satellites 11 , 12 , 13 , 14 in geostationary orbit, including a satellite 13 that is intended to communicate with an earth station 15 and several adjacent satellites 11 , 12 and 14 .","Satellite 13 transmits signals on a downlink band to the earth station 15 having a frequency down converter, A/D converter and CSM equipment 18 that can provide conventional CSM functions.","The CSM 18 has an adequate processing capability, which may be provided by a conventional processor (not shown) with appropriate software modules.","The CSM system 18 includes a digital spectrum analyzer, operating on the A/D output bits, that provides a panoramic look of the carriers in the entire frequency band that was digitized.","In order to properly monitor the received signal, the carriers must be separated or isolated, and subdivided as needed.","Then, an estimation made of their parameters, identifying their modulation types, as well as detecting and characterizing any interferer that may be present in the digitized frequency band.","The successive steps involve carrier isolation, segmentation, frequency estimation, symbol rate estimation, bit error rate estimation, modulation classification and interference characterization.","Carrier Isolation: Carrier isolation consists of identifying and separating the individual carriers in the digitized frequency band.","After carrier isolation has been performed, each carrier is processed separately to estimate its parameters, determine its modulation type, etc.","Typically, the carriers are identified on a spectrum analyzer by a human operator, in a straightforward and well known process.","However, automated carrier detection is difficult, as the process must be capable of differentiating true carriers from thermal noise, statistical fluctuations, side lobes, intermodulation products, and spurious spikes.","Segmentation: For the sake of computational simplicity, it is often necessary to segment the time domain data record containing the digital samples into segments of appropriate size, and to process each segment separately.","Furthermore, when the channel is not constant, segmentation has the additional advantage of providing a channel which is approximately constant over each segment.","Examples of non-constant channels include bursty channels, fading channels, and voice activated channels.","The size of the segment is usually chosen as a power of 2 because such a choice leads to the use of efficient FFT processing.","FFT processing is the backbone of the digital spectrum analysis to be performed on such segments.","Frequency Estimation: There are several well-known techniques to carrier frequency estimation.","One popular technique is the centroid method.","In this method, the center frequency is estimated as a weighted average frequency, where the weights are taken as the squares of the spectral coefficients.","A second method consists of fitting a straight line to the instantaneous phase data.","Finding the best straight-line fit is a simple mean square error minimization problem, where the slope of the line provides the frequency estimate and the value at the origin provides the initial phase.","When using this technique, the phase values must be unwrapped before the straight line fit A third method consists of passing the received waveform through a nonlinearity, such as quadrupling, and detecting spectral lines at harmonics of the carrier.","The frequency location of these spectral lines, which are obtained via a high resolution FET, would provide an accurate estimate of the carrier frequency.","While the above three methods are suitable in many situations, they each have their shortcomings, making them unsuitable for some applications.","For example, the centroid method is not suitable if the frequency spectrum is not symmetric.","The instantaneous phase square error minimization is best suited to constant envelope modulations, and the nonlinearity does not always produce line spectra at harmonics of the carrier frequency.","Furthermore, the accuracy provided by these methods may sometimes be insufficient.","Symbol Rate Estimation: There are several well-known techniques for symbol rate estimation.","One conventional scheme is the delay and multiply method, where the received waveform is multiplied by a replica of itself, that has been delayed by a fraction of the symbol rate.","Spectral lines will then appear in the spectrum at harmonics of the symbol rate when the delay is properly chosen.","The amount of delay, and the number and magnitude of spectral lines are modulation scheme-dependent and well known in each case.","Those spectral lines therefore provide a signature identifying the symbol rate, and may also be used for modulation discrimination.","Another method is to use a first order phase lock loop (PLL) to track the timing of the received signal.","This is a typical way of achieving clock synchronization in digital modems.","While the above methods are suitable for many situations, they each have their shortcomings, making them unsuitable for some applications.","For example, the delay and multiply method does not always produce spectral lines at harmonics of the symbol rate.","As to the PLL tracking method, it needs a sufficiently accurate knowledge of the symbol rate at the start.","BER Estimation: When a signal is demodulated and FEC is decoded, it is possible to obtain an accurate estimate of the BER, without having access to the actual transmitted bits.","BER is determined by a well-known procedure based on re-encoding the decoded bits.","In the absence of FEC decoding, an accurate BER estimate (coded or uncoded) may be obtained over an AWGN (additive white gaussian noise) channel from accurate estimation of energy per bit/noise density (Eb/No), and knowledge of the modulation format and FEC type and rate.","Estimating the uncoded and coded BER becomes more difficult if the channel is not AWGN.","In order to provide a fairly accurate BER estimate in this case, understanding the nature and magnitude of the various channel impairments is paramount.","Indeed, if by a process of reverse engineering one is able to completely determine all the channel impairments, then one could in principle reconstruct a waveform statistically identical to the one under examination, and therefore one would be able to accurately estimate the BER.","In reality of course, it is not possible to completely determine all the channel impairments, and one would attempt to estimate them as accurately as possible.","Modulation Classification: There are a number of well-know techniques for modulation classification.","They mostly fall into one of two categories: pattern-recognition based and decision theory-based.","The most practical techniques are a hybrid of these two approaches, where a set of key features is extracted from the modulated waveform (as in pattern recognition), and the principles of decision theory are applied to classify the modulation based on those features.","Numerous key features have been used for modulation classification.","A partial list of those features include: amplitude histograms, frequency histograms, phase histograms, phase difference histograms, the variance of the amplitude, frequency, and phase, higher order moments, kurtosis, cumulants, the square of the signal envelope, zero crossings, the power spectrum of the received signal, the presence of harmonics at selected frequencies, the magnitude of the spectral component at twice the carrier frequency of the signal squared, the magnitude of the spectral component at 4 times the carrier frequency of the signal raised to the fourth power, and the power spectrum asymmetry.","If properly chosen and applied, the key features can help discriminate among different modulation formats, even under adverse conditions, such as low signal to noise ratio (SNR), limited amount of data, presence of interference, and channel impairments.","Existing modulation classification schemes typically have several shortcomings.","One shortcoming is that the key features computation does not take into account that different samples have different reliability values, as they are often taken asynchronously with the signal symbols.","Another shortcoming is that the band-limited nature of the waveform (which causes signal fluctuations around the symbol edges) is usually not taken into account A third shortcoming is that the outcome of a classification scheme is often dependent on the sequence of applying the key features.","Another shortcoming is that the thresholds used in determining the decision regions are independent of SNR.","A further shortcoming is that simple majority rule is used to make a final decision based on the individual segments decisions.","Last but not least, is the fact that many existing classification schemes require exact knowledge of the signal parameters, and are not robust to inaccuracies in the value of those parameters.","Unfortunately, the schemes that perform the best under idealized conditions tend to be the least robust.","Interference Characterization: Interference identification and characterization can significantly enhance the utility of a Communication System Monitoring system.","In this regard, “characterization” refers to determining the power level; carrier frequency and occupied bandwidth of the interferer, plus any other transmission parameters that may be estimated.","Generalizing the interference characterization to the case of multiple interferers is done iteratively.","There are many potential sources of interference in a satellite communication system such as inclined satellites, radars, terrestrial microwave links, in-orbit test equipment generated carriers, rogue transmitters, and carriers on mistaken frequencies and/or directions.","In addition, as previously noted, adjacent satellites in the geostationary arc are a main source of interference.","Adjacent Satellite Interference (ASI) can occur on the uplink and on the downlink.","While the interference mechanism is different in these two cases, both uplink ASI and downlink ASI result in the presence of interfering signals in a frequency band.","An interferer's power may be sufficiently low to make its detection and identification difficult, yet sufficiently high to cause noticeable performance degradation to desired signals.","Furthermore, the capability to characterize interferers in a desired frequency band can provide useful data on whether other satellite systems are abiding by the frequency coordination agreements to which they are party.","A practical algorithm for interference identification and characterization is known in the art.","The received waveform consists of a distorted version of the desired signal, thermal noise, and an unknown interferer.","It is desired to characterize the interferer to the extent possible.","In other words, it is desired to determine the interferer power, center frequency, occupied bandwidth, modulation type, symbol rate, and any other potentially useful information.","If one could completely cancel out the desired signal, standard correlation techniques could be used to extract interferer information from the thermal noise.","However, the distortion of the desired signal makes its complete cancellation impractical.","The goal is then to cancel the desired signal as much as possible so that any residual energy is small and does not mask the presence of an interferer.","Impairments that are expected to distort the desired signal waveform include: the non-ideal channel, phase noise, oscillator drift, transmitter non-linearities, non-ideal filtering, clock jitter, intermodulation products, and quadrature imbalance.","In order to perform a fairly complete cancellation of the strong signal in this case, understanding the nature and magnitude of the various channel impairments is paramount.","Indeed, if by a process of reverse engineering one is able to completely determine all the channel impairments, then one could in principle reconstruct a noise-free, identical copy of the desired signal in the received waveform.","Subtracting this constructed replica from the received waveform would leave the interferer and the noise.","In reality of course, it is not possible to construct a perfect noise-free copy of the desired signal in the received waveform.","The desired approach is to construct as close a replica as possible of the received desired signal by estimating the impairments as accurately as possible.","The extent to which it is possible to estimate those impairments and cancel out their effect will determine the degree of success in characterizing the interference.","As many of the foregoing processes and procedures are manual or only semi-automated, it is an object of the present invention to provide fully automated procedures for determining each of these satellite performance related parameters.","It is also an object of the invention to provide a combination of automated procedures that can attain an automatic generation of satellite frequency plans.","It is yet an object of the invention to provide a combination of at least two and possibly all of the automated procedures in order to obtain an optimum result."],["<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is a digital signal processing (DSP)-based approach to parameter estimation, modulation identification and interference characterization in connection with a satellite Communication Monitoring System (CSM).","The techniques described here allow automatic generation of satellite frequency plans without any a priori knowledge of such plans.","When combined with information publicly available about a given satellite, these techniques will give very precise information of the frequency plan of that satellite."],["This application claims the benefit of U.S.","Provisional Application Ser.","No.","60/289,389, filed on May 8, 2002.FIELD OF THE INVENTION The invention relates generally to a satellite communication monitoring (CSM) method and apparatus for providing parameter estimation, modulation classification, and interference characterization in communication satellite systems.","BACKGROUND In a satellite communication system, particularly a system where the satellite is deployed in a geostationary orbit, the satellite will be able to receive signals transmitted to the satellite by earth stations at an allocated uplink frequency band and will be operative to transmit signals to earth stations on allocated downlink frequency bands.","The uplink bands are selected to be spaced apart from the downlink bands in order to avoid interference.","Nonetheless, interference may be generated due to transmissions from adjacent earth stations or adjacent satellites having overlapping beams.","In addition, interference may arise from natural phenomena, such as rainfall, scattering, terrestrial communications and the like.","With respect to the downlink, the signal received at an earth station from the satellite is frequency-down-converted and digitized by means of an analog-to-digital (A/D) converter.","A typical value for the bandwidth of the A/D converted frequency band is 36 MHz.","The A/D converter output is a stream of bits which essentially captures all the information in the received signal.","FIG.","1 illustrates a satellite system 10 having a plurality of satellites 11, 12, 13, 14 in geostationary orbit, including a satellite 13 that is intended to communicate with an earth station 15 and several adjacent satellites 11, 12 and 14.Satellite 13 transmits signals on a downlink band to the earth station 15 having a frequency down converter, A/D converter and CSM equipment 18 that can provide conventional CSM functions.","The CSM 18 has an adequate processing capability, which may be provided by a conventional processor (not shown) with appropriate software modules.","The CSM system 18 includes a digital spectrum analyzer, operating on the A/D output bits, that provides a panoramic look of the carriers in the entire frequency band that was digitized.","In order to properly monitor the received signal, the carriers must be separated or isolated, and subdivided as needed.","Then, an estimation made of their parameters, identifying their modulation types, as well as detecting and characterizing any interferer that may be present in the digitized frequency band.","The successive steps involve carrier isolation, segmentation, frequency estimation, symbol rate estimation, bit error rate estimation, modulation classification and interference characterization.","Carrier Isolation: Carrier isolation consists of identifying and separating the individual carriers in the digitized frequency band.","After carrier isolation has been performed, each carrier is processed separately to estimate its parameters, determine its modulation type, etc.","Typically, the carriers are identified on a spectrum analyzer by a human operator, in a straightforward and well known process.","However, automated carrier detection is difficult, as the process must be capable of differentiating true carriers from thermal noise, statistical fluctuations, side lobes, intermodulation products, and spurious spikes.","Segmentation: For the sake of computational simplicity, it is often necessary to segment the time domain data record containing the digital samples into segments of appropriate size, and to process each segment separately.","Furthermore, when the channel is not constant, segmentation has the additional advantage of providing a channel which is approximately constant over each segment.","Examples of non-constant channels include bursty channels, fading channels, and voice activated channels.","The size of the segment is usually chosen as a power of 2 because such a choice leads to the use of efficient FFT processing.","FFT processing is the backbone of the digital spectrum analysis to be performed on such segments.","Frequency Estimation: There are several well-known techniques to carrier frequency estimation.","One popular technique is the centroid method.","In this method, the center frequency is estimated as a weighted average frequency, where the weights are taken as the squares of the spectral coefficients.","A second method consists of fitting a straight line to the instantaneous phase data.","Finding the best straight-line fit is a simple mean square error minimization problem, where the slope of the line provides the frequency estimate and the value at the origin provides the initial phase.","When using this technique, the phase values must be unwrapped before the straight line fit A third method consists of passing the received waveform through a nonlinearity, such as quadrupling, and detecting spectral lines at harmonics of the carrier.","The frequency location of these spectral lines, which are obtained via a high resolution FET, would provide an accurate estimate of the carrier frequency.","While the above three methods are suitable in many situations, they each have their shortcomings, making them unsuitable for some applications.","For example, the centroid method is not suitable if the frequency spectrum is not symmetric.","The instantaneous phase square error minimization is best suited to constant envelope modulations, and the nonlinearity does not always produce line spectra at harmonics of the carrier frequency.","Furthermore, the accuracy provided by these methods may sometimes be insufficient.","Symbol Rate Estimation: There are several well-known techniques for symbol rate estimation.","One conventional scheme is the delay and multiply method, where the received waveform is multiplied by a replica of itself, that has been delayed by a fraction of the symbol rate.","Spectral lines will then appear in the spectrum at harmonics of the symbol rate when the delay is properly chosen.","The amount of delay, and the number and magnitude of spectral lines are modulation scheme-dependent and well known in each case.","Those spectral lines therefore provide a signature identifying the symbol rate, and may also be used for modulation discrimination.","Another method is to use a first order phase lock loop (PLL) to track the timing of the received signal.","This is a typical way of achieving clock synchronization in digital modems.","While the above methods are suitable for many situations, they each have their shortcomings, making them unsuitable for some applications.","For example, the delay and multiply method does not always produce spectral lines at harmonics of the symbol rate.","As to the PLL tracking method, it needs a sufficiently accurate knowledge of the symbol rate at the start.","BER Estimation: When a signal is demodulated and FEC is decoded, it is possible to obtain an accurate estimate of the BER, without having access to the actual transmitted bits.","BER is determined by a well-known procedure based on re-encoding the decoded bits.","In the absence of FEC decoding, an accurate BER estimate (coded or uncoded) may be obtained over an AWGN (additive white gaussian noise) channel from accurate estimation of energy per bit/noise density (Eb/No), and knowledge of the modulation format and FEC type and rate.","Estimating the uncoded and coded BER becomes more difficult if the channel is not AWGN.","In order to provide a fairly accurate BER estimate in this case, understanding the nature and magnitude of the various channel impairments is paramount.","Indeed, if by a process of reverse engineering one is able to completely determine all the channel impairments, then one could in principle reconstruct a waveform statistically identical to the one under examination, and therefore one would be able to accurately estimate the BER.","In reality of course, it is not possible to completely determine all the channel impairments, and one would attempt to estimate them as accurately as possible.","Modulation Classification: There are a number of well-know techniques for modulation classification.","They mostly fall into one of two categories: pattern-recognition based and decision theory-based.","The most practical techniques are a hybrid of these two approaches, where a set of key features is extracted from the modulated waveform (as in pattern recognition), and the principles of decision theory are applied to classify the modulation based on those features.","Numerous key features have been used for modulation classification.","A partial list of those features include: amplitude histograms, frequency histograms, phase histograms, phase difference histograms, the variance of the amplitude, frequency, and phase, higher order moments, kurtosis, cumulants, the square of the signal envelope, zero crossings, the power spectrum of the received signal, the presence of harmonics at selected frequencies, the magnitude of the spectral component at twice the carrier frequency of the signal squared, the magnitude of the spectral component at 4 times the carrier frequency of the signal raised to the fourth power, and the power spectrum asymmetry.","If properly chosen and applied, the key features can help discriminate among different modulation formats, even under adverse conditions, such as low signal to noise ratio (SNR), limited amount of data, presence of interference, and channel impairments.","Existing modulation classification schemes typically have several shortcomings.","One shortcoming is that the key features computation does not take into account that different samples have different reliability values, as they are often taken asynchronously with the signal symbols.","Another shortcoming is that the band-limited nature of the waveform (which causes signal fluctuations around the symbol edges) is usually not taken into account A third shortcoming is that the outcome of a classification scheme is often dependent on the sequence of applying the key features.","Another shortcoming is that the thresholds used in determining the decision regions are independent of SNR.","A further shortcoming is that simple majority rule is used to make a final decision based on the individual segments decisions.","Last but not least, is the fact that many existing classification schemes require exact knowledge of the signal parameters, and are not robust to inaccuracies in the value of those parameters.","Unfortunately, the schemes that perform the best under idealized conditions tend to be the least robust.","Interference Characterization: Interference identification and characterization can significantly enhance the utility of a Communication System Monitoring system.","In this regard, “characterization” refers to determining the power level; carrier frequency and occupied bandwidth of the interferer, plus any other transmission parameters that may be estimated.","Generalizing the interference characterization to the case of multiple interferers is done iteratively.","There are many potential sources of interference in a satellite communication system such as inclined satellites, radars, terrestrial microwave links, in-orbit test equipment generated carriers, rogue transmitters, and carriers on mistaken frequencies and/or directions.","In addition, as previously noted, adjacent satellites in the geostationary arc are a main source of interference.","Adjacent Satellite Interference (ASI) can occur on the uplink and on the downlink.","While the interference mechanism is different in these two cases, both uplink ASI and downlink ASI result in the presence of interfering signals in a frequency band.","An interferer's power may be sufficiently low to make its detection and identification difficult, yet sufficiently high to cause noticeable performance degradation to desired signals.","Furthermore, the capability to characterize interferers in a desired frequency band can provide useful data on whether other satellite systems are abiding by the frequency coordination agreements to which they are party.","A practical algorithm for interference identification and characterization is known in the art.","The received waveform consists of a distorted version of the desired signal, thermal noise, and an unknown interferer.","It is desired to characterize the interferer to the extent possible.","In other words, it is desired to determine the interferer power, center frequency, occupied bandwidth, modulation type, symbol rate, and any other potentially useful information.","If one could completely cancel out the desired signal, standard correlation techniques could be used to extract interferer information from the thermal noise.","However, the distortion of the desired signal makes its complete cancellation impractical.","The goal is then to cancel the desired signal as much as possible so that any residual energy is small and does not mask the presence of an interferer.","Impairments that are expected to distort the desired signal waveform include: the non-ideal channel, phase noise, oscillator drift, transmitter non-linearities, non-ideal filtering, clock jitter, intermodulation products, and quadrature imbalance.","In order to perform a fairly complete cancellation of the strong signal in this case, understanding the nature and magnitude of the various channel impairments is paramount.","Indeed, if by a process of reverse engineering one is able to completely determine all the channel impairments, then one could in principle reconstruct a noise-free, identical copy of the desired signal in the received waveform.","Subtracting this constructed replica from the received waveform would leave the interferer and the noise.","In reality of course, it is not possible to construct a perfect noise-free copy of the desired signal in the received waveform.","The desired approach is to construct as close a replica as possible of the received desired signal by estimating the impairments as accurately as possible.","The extent to which it is possible to estimate those impairments and cancel out their effect will determine the degree of success in characterizing the interference.","As many of the foregoing processes and procedures are manual or only semi-automated, it is an object of the present invention to provide fully automated procedures for determining each of these satellite performance related parameters.","It is also an object of the invention to provide a combination of automated procedures that can attain an automatic generation of satellite frequency plans.","It is yet an object of the invention to provide a combination of at least two and possibly all of the automated procedures in order to obtain an optimum result.","SUMMARY OF THE INVENTION The present invention is a digital signal processing (DSP)-based approach to parameter estimation, modulation identification and interference characterization in connection with a satellite Communication Monitoring System (CSM).","The techniques described here allow automatic generation of satellite frequency plans without any a priori knowledge of such plans.","When combined with information publicly available about a given satellite, these techniques will give very precise information of the frequency plan of that satellite.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is a schematic illustration of a satellite system where CSM would be utilized.","FIG.","2 is a flowchart for a procedure that performs an automated carrier isolation process.","FIG.","3 is a flowchart for a procedure that performs an automated carrier segmentation process.","FIG.","4 is a flowchart for a procedure that performs an automated carrier frequency estimation process.","FIG.","5 is a flowchart for a procedure that performs an automated symbol rate estimation process.","FIG.","6 is a flowchart for a procedure that performs an automated bit error rate (BER) estimation process.","FIG.","7 is a flowchart for a procedure that performs an automated modulation classification process.","FIG.","8 is a flowchart for a procedure that performs an automated interference identification and characterization process.","FIG.","9A is a flowchart for a procedure that integrates three of the foregoing automated processes to achieve an automatic generation of a satellite frequency plan.","FIG.","9B is a flowchart for a procedure that integrates all of the foregoing automated processes.","DETAILED DESCRIPTION OF THE INVENTION In a satellite communication system as illustrated in FIG.","1, where a CSM system is available to perform various system monitoring, analysis and estimation functions, the processor resident at the CSM may be operative to perform a variety of procedures, consistent with the algorithms identified subsequently in flowchart form, to automatically estimate parameters, classify modulation and characterize interference in the system.","While the invention is disclosed in connection with various specific embodiments, it is not limited thereto and a wide variety of alternative approaches may be evident to one skilled in the art upon reading the following disclosure.","For example, the processing performed may be distributed or centralized, with communication with an earth station provided by well known network and system arrangements.","Turning now to the individual elements of the signal processing performed in a CSM system contemplated by the present invention, the following procedures may be employed.","Carrier Isolation: Carrier isolation would be automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.","2.In a first step S1, the processor serving the CSM would perform an FFT of size NFFT on the composite waveform of bandwidth W. The value of NFFT is programmable, and may be given a default value, such as but not limited to 1024.In step S2, the power spectrum is obtained from the FFT output by computing the squared magnitude of each FFT coefficient.","This procedure is repeated M times according to step S3 and averaged out in step S4 to smooth out the statistical fluctuations in the spectrum.","The value of M is programmable, and in an exemplary embodiment, may have a default value of 16, although other values would be readily apparent to one skilled in the art.","Then, a noise floor pn is determined in step S5.With these basic parameters in hand, the power spectrum is filtered in step S6 in order to mitigate the impact of any statistical fluctuations, and to gloss over spurious spikes and frequency nulls between sidelobes.","Then, in step S7, a minimum carrier level pc X dB above the noise floor pn is set, where X is a programmable parameter, which may have a default value of 3 dB, or other value as would be apparent to one skilled in the art.","The processing will proceed through the individual frequency points from the lowest to the highest, in step S8.When a value higher than pc is first detected at a certain frequency, that frequency is taken as the lower frequency limit of a carrier in step S8.Also, in that same step, when the value at the filter output drops first to a level below pc, the corresponding frequency is taken as the upper frequency limit of the carrier.","The entire spectrum is processed according to the procedure of step S8, as indicated in step S9, thus identifying the lower and upper frequency limits for each carrier.","Finally, in step S10, the individual carriers are digitally filtered out, one by one, in accordance with the procedure identified above.","Thereafter, the procedure comes to an end.","Segmentation: Segmentation would be automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.","3.In a first step S11, a number of samples that are to be included in a segment are determined and a suitable power of 2 is identified to represent the selected number of samples in the segment.","Then, in step S12, the instantaneous power in each sample is computed.","Next, in step S13, the instantaneous power values are filtered out in order to remove large deviations.","In step S14, the average and the standard deviation of the instantaneous power is computed and, in step S15, the normalized standard deviation is computed and compared to a threshold based on the value of Eb/No, where Eb is the signal strength and No is a corresponding noise value.","In step S16, it is determined whether the threshold is exceeded.","If not exceeded, the segment is rejected (N), otherwise (Y), it is accepted in step S17.Then in step S18, it is determined whether the analyzed segment is the last segment and, if not, the process proceeds to test the next segment If it is the last segment, the process ends.","Frequency Estimation: Frequency estimation is automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.","4.In a first step S21, the processor would compute a frequency estimate using the well known centroid method.","Then, in step S22, the instantaneous phase method is modified.","The modification would be made as follows.","Assign a weight proportional to the square of the magnitude to each instantaneous phase value, so that samples with higher SNR are given more weight.","Use a weighted minimum mean square error criterion.","Then, in step S23, a second frequency estimate is computed using the modified instantaneous phase method as described above.","In step S24, the waveform is passed through a non-linearity and any harmonics in the spectrum are detected.","In step S25, the FFT-based location of harmonics is enhanced by using unbiased interpolation of the FFT coefficients.","Finally, in step S26, a third frequency estimate is determined on the basis of the enhanced harmonics location process, as previously disclosed.","Once the three frequency estimates are obtained, although more may be obtained if desired, a weighted average of the frequency estimates is determined in step S27.The weights are assigned on the basis of spectral symmetry, envelope fluctuations, and strength of the frequency harmonics.","As would be understood by one skilled in the art, if only a moderate frequency estimation accuracy is sought, a subset of the above set of estimates would be adequate.","On the other hand, if higher frequency accuracy is still needed, supplement the estimate obtained above with a phase locked loop to track the received carrier.","Symbol Rate Estimation: Symbol rate estimation is automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.","5, using a modification of the conventional delay and multiply technique.","According to the modification, in a step S31, both the received signal and its delayed replica are passed through a nonlinearity to produce harmonics at the symbol rate.","Then, in step S32, the processor would compute the number of crossings per unit time where the signal envelope crosses the half power level.","In step S33, the timing (clock frequency and phase) of the received waveform would be tracked, in an exemplary embodiment, by using a second order PLL.","In a subsequent process represented by step S34, a non-uniform sampling approach would be used.","For example, but without limitation, a non-uniformly sampled set may be generated by digital interpolation between the available uniformly sampled samples.","The proposed non-uniform sampling rate is slowly and monotonically increasing, and covers the range of uncertainty in the symbol rate.","This provides the ability to home in on the true sample rate.","Once lock is achieved, uniform sampling is resumed and a PLL is used to fine tune the symbol rate estimate.","If only a moderate symbol rate estimation accuracy is sought, a subset of the above set of estimates would be adequate.","BER Estimation: Bit error rate estimation is automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.","6.According to step S41, the process begins with an estimate the waveform parameters and a determine the modulation type, if it is not already known, as described above.","Then, in step S42, there is a processing of the received samples with a properly matched and equalized filter, followed by a tracking of the carrier phase and the clock phase in step S43.In step S44, the well known maximum likelihood techniques are used to estimate the phase noise, intermodulation products, quadrature imbalances, and non-linearity's.","Any side information available regarding the transmitter characteristics, such as for example the power amplifier specifications, may be used for this purpose in step S45.The information may be available beforehand and either input manually or accessible automatically by the processor on the basis of pre-stored information in RAM or auto detected characteristics of the equipment, in a manner known in the art.","The process proceeds in option 1 to the construction of a waveform with the estimated parameters and modulation type, subject it to the estimated impairments, and estimate the BER, in step S46.Alternatively, the process may proceed as option 2 to step S47 by first constructing a noise-free scattering diagram based on the estimated impairments.","Then, an estimate of the uncoded and coded BER, using maximum likelihood, may be obtained from the noise-free scattering diagram, and the estimated Eb/No in step S48.Modulation Classification: Modulation classification is automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.","7.According to step S51, an estimate is automatically made of the waveform parameters as accurately as possible, as outlined above.","Then, an estimate of the signal-to-noise ratio (SNR) of the received waveform is obtained in step S52.Available side information, if any, may be used to narrow down the set of potential modulation formats at this point, according to step S53.The information may be available beforehand and either input manually or accessible automatically by the processor on the basis of pre-stored information in RAM or auto detected characteristics of the equipment, in a manner known in the art.","Then several modification steps occur.","In step S54, the key features computation is modified such that each sample contributing to a key feature is assigned a weight proportional to its SNR.","(Some phase samples are more sensitive to noise than others, depending on the magnitude of those samples.)","In step S55, the key features computation is modified such that each sample contributing to a key feature is assigned a weight proportional to its distance from the symbol edges.","(Band limiting causes envelope fluctuations around the symbol edges).","In step S56, based on the side information, a subset of key features from the set listed above is computed.","Then, in step S57, the sub-optimum hierarchical classification approach to a vector approach is modified, where several features are applied simultaneously to a multidimensional threshold.","The threshold setting is made SNR-dependent.","(Actual threshold values for different SNRs are computed offline).","In step S58, the number of segments processed is made SNR-dependent to achieve a given confidence level.","And, in step S59, for each segment processed, a ranking is assigned as to how likely it is that the waveform under examination belongs to each of the modulation classes under consideration.","Finally, in step S60, a soft combining of all the segment rankings is performed to arrive at the most likely overall classification of a modulation type.","Interference Characterization: Interference characterization is automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.","8.According to step S61, an estimate is made of the waveform parameters and a determination is made of the modulation type of the desired signal, if it is not already known, as described earlier.","In step S62, the received samples are processed with a properly matched and equalized filter.","Then, in step S63, the carrier phase and the clock phase are tracked.","Any side information available regarding the transmitter characteristics, such as for example the power amplifier specifications, may be used in this regard and optionally input.","The information may be available beforehand and either input manually or accessible automatically by the processor in step S63A on the basis of pre-stored information in RAM or auto detected characteristics of the equipment, in a manner known in the art.","In step S64, the well known maximum likelihood techniques is used to estimate the phase noise, intermodulation products, quadrature imbalances, and nonlinearities.","Then, in step S65, the received signal is demodulated and the transmitted bits are recovered.","Optionally, if the SNR is low and the error rate is high, FEC decoding of the signal to recover the information bits can be beneficial in this step.","If FEC decoding was performed, the information bits must be re-encoded.","In step S66, the transmitted bits are remodulated on a carrier according to the known (or estimated) modulation type, symbol rate, and filter characteristics.","The remodulated signal is subjected to the impairments estimated above in step S67 and the remodulated signal from the received waveform in step S68.A standard correlation and spectral analysis techniques is performed on the residual signal to extract interferer information from the noise, in step S69.The several processes for automated determination of parameters may be combined to provide an automatic generation of a satellite frequency plan, as illustrated in FIG.","1, and according to the process of FIG.","9A.","Specifically, the carrier is isolated automatically, according to the process in FIG.","2, in step S71.The carrier isolation is followed by a segmentation processing in step S72, according to the flowchart of FIG.","3.Thereafter, a frequency estimation process according to the method of FIG.","4 is conducted automatically in step S73.The result of this combination of outputs would be automatically combined into a frequency plan for a satellite by the CSM in step S74.The several processes disclosed in FIGS.","2-8 may be conducted automatically, in any combination, as would be known in the art, including a combination of all of the processes as illustrated in FIG.","9B.","There, as in FIG.","9A, the carrier isolation, segmentation and frequency estimation processes, which derive a frequency plan in step 81, may be accompanied by the estimation of symbol rate according to the process of FIG.","5 in step S82 and the estimation of BER according to the process of FIG.","6 in step S83 The modulation classification according to FIG.","7 may be performed in step S84 and the interference characterization according to FIG.","8 may be performed in step S85.While the present invention has been described in accordance with certain embodiments and examples, it is not limited thereto."]],"string":"[\n [\n \"Method and apparatus for parameter estimation, modulation classification and interference characterization in satellite communication systems\",\n \"A digital signal processing (DSP)-based approach to parameter estimation modulation identification and interference charaterization in connection with a satellite Communication Monitoring System (CSM).\",\n \"The techniques descried here also allow automatic generation of satellite frequency plans (S74) without any a priori knowledge of such plans.\",\n \"Individual processes for carrier isolation (S71), segmentation (S72), frequency estimation (S73), symbol rate estimation, bit error rate estimation, modulation identification and interference characterization are disclosed may be combined in a totally automated process.\"\n ],\n [\n \"1.A method of automatically isolating carriers of a composite waveform having a bandwidth in a satellite communication system, comprising: a) performing an FFT processing of size NFFT on the composite waveform, where the value of NFFT is programmable; b) obtaining a power spectrum from the FFT processing by computing the squared magnitude of each FFT coefficient; c) repeating step b) a plurality of times and averaging the results; d) setting, a noise floor pn; e) filtering the power spectrum; f) setting a minimum carrier level pc X dB above the noise floor pn, where X is a programmable parameter, g) identifying the lower and upper frequency limits for each carrier; and h) digitally filtering the individual carriers.\",\n \"2.The method of claim 1 wherein, when a value higher than pc is first detected at a certain frequency, that frequency is taken as the lower frequency limit of a carrier and when the value at the filter output drops first to a level below pc, the corresponding frequency is taken as the upper frequency limit of the carrier.\",\n \"3.The method of claim 1 wherein.\",\n \"The processing proceeds through individual frequency points within the spectrum.\",\n \"4.A method of automatically providing segmentation of time domain data record in a satellite communication system comprising: a) determining a number of samples that are to be included in a segment; b) computing, the instantaneous power in each sample; c) filtering instantaneous power values; d) computing an average and a standard deviation of the instantaneous power; e) computing a normalized standard deviation and comparing the normalized standard deviation to a threshold; f) determining whether the threshold is exceeded and if not exceeded, the segment is rejected, otherwise, it is accepted; and g) repeating the foregoing process for at least one additional segment.\",\n \"5.The method of claim 4 wherein the determining step includes identifying a suitable power of 2 to represent the selected number of samples in the segment.\",\n \"6.The method of claim 4 wherein the threshold of step e) is based on a value of Eb/No, where Eb is the signal strength and No is a corresponding noise value.\",\n \"7.A method of automatically estimating frequency in a satellite communication system comprising: a) computing a frequency estimate; b) providing a modified an instantaneous phase method; c) computing a second frequency estimate is computed using the modified instantaneous phase method; d) detecting any harmonics in the spectrum; e) enhancing a FFT-based location of harmonics by using unbiased interpolation of the FFT coefficients; f) computing a third frequency estimate on the basis of the enhanced harmonics location process; g) determining a weighted average of the frequency estimates; h) assigning weights on the basis of spectral symmetry, envelope fluctuations, and strength of the frequency harmonics.\",\n \"8.The method of claim 7 wherein step a) uses the centroid method.\",\n \"9.The method of claim 7 wherein the modifying step comprises assigning a weight proportional to the square of the magnitude to each instantaneous phase value, so that samples with higher SNR are given more weight.\",\n \"10.The method of claim 7 wherein the waveform is passed through a non-linearity 11.The method of claim 7 further comprising using a phase locked loop (PLL) to track the received carrier.\",\n \"12.A method of automatically estimating symbol rate in a satellite communication system comprising: a) applying a delay and multiply technique wherein both a received signal and its delayed replica are passed through a non-linearity to produce harmonics at the symbol rate; b) computing the number of crossings per unit time where a signal envelope crosses a half power level; c) tracking the timing of the received waveform; d) providing non-uniform sampling at a non-uniform sampling rate that is slowly and monotonically increasing, and covers the range of uncertainty in the symbol rate; e) once lock is achieved, resuming uniform sampling; and f) using a PLL to fine tune the symbol rate estimate.\",\n \"13.The method of claim 12 wherein the timing step comprises tracking clock frequency and phase, using a second order PLL.\",\n \"14.The method of claim 12 wherein said non-uniform sampling comprises generating a set by digital interpolation between the available uniformly sampled samples.\",\n \"15.A method of automatically estimating bit error rate on received signals in a satellite communication system, comprising: a) processing the received signals with a properly matched and equalized filter; b) tracking of the carrier phase and the clock phase; c) using maximum likelihood techniques to estimate one or more of the phase noise, intermodulation products, quadrature imbalances, and non-linearity's.\",\n \"d) constructing a waveform with estimated parameters and modulation type; e) subjecting the waveform to estimated impairments, f) estimating the bit error rate.\",\n \"16.The method of claim 15 further comprising, estimating the waveform parameters and determining the modulation type.\",\n \"17.The method of claim 15 further comprising automatically considering side information regarding the transmitter characteristics.\",\n \"18.The method of claim 15 further comprising constructing a noise-free scattering diagram based on the estimated impairments, estimating the uncoded and coded BER, using maximum likelihood, from the noise-free scattering diagram, and obtaining an estimated Eb/No, where Eb is the signal strength and No is a corresponding noise value.\",\n \"19.A method of automatically classifying modulation of a received signal in a satellite communication system, comprising: a) estimating the parameters of the received signal waveform; b) estimating the signal-to-noise ratio (SNR) of the received signal waveform; c) assigning each sample contributing to a key feature a weight proportional to its SNR; d) modifying the key features computation such that each sample contributing to a key feature is assigned a weight proportional to its distance from the symbol edges; e) modifying a sub-optimum hierarchical classification approach to a vector approach, wherein several features are applied simultaneously to a multidimensional threshold; f) making the number of segments processed SNR-dependent; g) for each segment processed, assign a ranking as to how likely it is that the waveform under examination belongs to each of the modulation classes under consideration; and h) soft combining all the segment rankings to arrive at the most likely overall classification of a modulation type.\",\n \"20.The method of claim 19, further comprising using side information to narrow down the set of potential modulation formats.\",\n \"21.The method of claim 19 wherein, in the modifying step e) the threshold setting is made SNR-dependent; 22.The method of claim 19 wherein the side information is input automatically from a data base.\",\n \"23.A method of automatically characterizing interference in a satellite communication system comprising: a) obtaining waveform parameters and modulation type of the desired signal, b) processing the received samples with a properly matched and equalized filter; c) tracking the carrier phase and the clock phase; d) estimating at least one of the phase noise, intermodulation products, quadrature imbalances, and non-linearity's using maximum likelihood techniques; e) demodulating the received signal and recovering the transmitted bits; f) remodulating the transmitted bits on a carrier according to the modulation type, symbol rate, and filter characteristics; and g) performing a correlation and spectral analysis on the residual signal to extract interferer information from the noise.\",\n \"24.The method of claim 23 further comprising estimating the parameters.\",\n \"25.The method of claim 23 further comprising and automatically determining modulation type.\",\n \"26.The method of claim 23 further comprising using side information available regarding the transmitter characteristics.\",\n \"27.The method of claim 23 wherein, if the SNR is low and the error rate is high, applying FEC decoding of the signal to recover the information bits; and re-encoding the information bits.\",\n \"28.A method of automatically generating a satellite frequency plan in a satellite system based on signals received from a satellite, comprising: a) isolating the carrier is isolated automatically; b) segmentation processing the received signal; and c) automatically estimating the frequency d) automatically combining the result of carrier isolation, segmentation and frequency estimation to develop a frequency plan for the satellite.\",\n \"29.An automated CSM system for use in a satellite communication system and operative to implement any one of the methods set forth in claim 1.\"\n ],\n [\n \"<SOH> BACKGROUND <EOH>In a satellite communication system, particularly a system where the satellite is deployed in a geostationary orbit, the satellite will be able to receive signals transmitted to the satellite by earth stations at an allocated uplink frequency band and will be operative to transmit signals to earth stations on allocated downlink frequency bands.\",\n \"The uplink bands are selected to be spaced apart from the downlink bands in order to avoid interference.\",\n \"Nonetheless, interference may be generated due to transmissions from adjacent earth stations or adjacent satellites having overlapping beams.\",\n \"In addition, interference may arise from natural phenomena, such as rainfall, scattering, terrestrial communications and the like.\",\n \"With respect to the downlink, the signal received at an earth station from the satellite is frequency-down-converted and digitized by means of an analog-to-digital (A/D) converter.\",\n \"A typical value for the bandwidth of the A/D converted frequency band is 36 MHz.\",\n \"The A/D converter output is a stream of bits which essentially captures all the information in the received signal.\",\n \"FIG.\",\n \"1 illustrates a satellite system 10 having a plurality of satellites 11 , 12 , 13 , 14 in geostationary orbit, including a satellite 13 that is intended to communicate with an earth station 15 and several adjacent satellites 11 , 12 and 14 .\",\n \"Satellite 13 transmits signals on a downlink band to the earth station 15 having a frequency down converter, A/D converter and CSM equipment 18 that can provide conventional CSM functions.\",\n \"The CSM 18 has an adequate processing capability, which may be provided by a conventional processor (not shown) with appropriate software modules.\",\n \"The CSM system 18 includes a digital spectrum analyzer, operating on the A/D output bits, that provides a panoramic look of the carriers in the entire frequency band that was digitized.\",\n \"In order to properly monitor the received signal, the carriers must be separated or isolated, and subdivided as needed.\",\n \"Then, an estimation made of their parameters, identifying their modulation types, as well as detecting and characterizing any interferer that may be present in the digitized frequency band.\",\n \"The successive steps involve carrier isolation, segmentation, frequency estimation, symbol rate estimation, bit error rate estimation, modulation classification and interference characterization.\",\n \"Carrier Isolation: Carrier isolation consists of identifying and separating the individual carriers in the digitized frequency band.\",\n \"After carrier isolation has been performed, each carrier is processed separately to estimate its parameters, determine its modulation type, etc.\",\n \"Typically, the carriers are identified on a spectrum analyzer by a human operator, in a straightforward and well known process.\",\n \"However, automated carrier detection is difficult, as the process must be capable of differentiating true carriers from thermal noise, statistical fluctuations, side lobes, intermodulation products, and spurious spikes.\",\n \"Segmentation: For the sake of computational simplicity, it is often necessary to segment the time domain data record containing the digital samples into segments of appropriate size, and to process each segment separately.\",\n \"Furthermore, when the channel is not constant, segmentation has the additional advantage of providing a channel which is approximately constant over each segment.\",\n \"Examples of non-constant channels include bursty channels, fading channels, and voice activated channels.\",\n \"The size of the segment is usually chosen as a power of 2 because such a choice leads to the use of efficient FFT processing.\",\n \"FFT processing is the backbone of the digital spectrum analysis to be performed on such segments.\",\n \"Frequency Estimation: There are several well-known techniques to carrier frequency estimation.\",\n \"One popular technique is the centroid method.\",\n \"In this method, the center frequency is estimated as a weighted average frequency, where the weights are taken as the squares of the spectral coefficients.\",\n \"A second method consists of fitting a straight line to the instantaneous phase data.\",\n \"Finding the best straight-line fit is a simple mean square error minimization problem, where the slope of the line provides the frequency estimate and the value at the origin provides the initial phase.\",\n \"When using this technique, the phase values must be unwrapped before the straight line fit A third method consists of passing the received waveform through a nonlinearity, such as quadrupling, and detecting spectral lines at harmonics of the carrier.\",\n \"The frequency location of these spectral lines, which are obtained via a high resolution FET, would provide an accurate estimate of the carrier frequency.\",\n \"While the above three methods are suitable in many situations, they each have their shortcomings, making them unsuitable for some applications.\",\n \"For example, the centroid method is not suitable if the frequency spectrum is not symmetric.\",\n \"The instantaneous phase square error minimization is best suited to constant envelope modulations, and the nonlinearity does not always produce line spectra at harmonics of the carrier frequency.\",\n \"Furthermore, the accuracy provided by these methods may sometimes be insufficient.\",\n \"Symbol Rate Estimation: There are several well-known techniques for symbol rate estimation.\",\n \"One conventional scheme is the delay and multiply method, where the received waveform is multiplied by a replica of itself, that has been delayed by a fraction of the symbol rate.\",\n \"Spectral lines will then appear in the spectrum at harmonics of the symbol rate when the delay is properly chosen.\",\n \"The amount of delay, and the number and magnitude of spectral lines are modulation scheme-dependent and well known in each case.\",\n \"Those spectral lines therefore provide a signature identifying the symbol rate, and may also be used for modulation discrimination.\",\n \"Another method is to use a first order phase lock loop (PLL) to track the timing of the received signal.\",\n \"This is a typical way of achieving clock synchronization in digital modems.\",\n \"While the above methods are suitable for many situations, they each have their shortcomings, making them unsuitable for some applications.\",\n \"For example, the delay and multiply method does not always produce spectral lines at harmonics of the symbol rate.\",\n \"As to the PLL tracking method, it needs a sufficiently accurate knowledge of the symbol rate at the start.\",\n \"BER Estimation: When a signal is demodulated and FEC is decoded, it is possible to obtain an accurate estimate of the BER, without having access to the actual transmitted bits.\",\n \"BER is determined by a well-known procedure based on re-encoding the decoded bits.\",\n \"In the absence of FEC decoding, an accurate BER estimate (coded or uncoded) may be obtained over an AWGN (additive white gaussian noise) channel from accurate estimation of energy per bit/noise density (Eb/No), and knowledge of the modulation format and FEC type and rate.\",\n \"Estimating the uncoded and coded BER becomes more difficult if the channel is not AWGN.\",\n \"In order to provide a fairly accurate BER estimate in this case, understanding the nature and magnitude of the various channel impairments is paramount.\",\n \"Indeed, if by a process of reverse engineering one is able to completely determine all the channel impairments, then one could in principle reconstruct a waveform statistically identical to the one under examination, and therefore one would be able to accurately estimate the BER.\",\n \"In reality of course, it is not possible to completely determine all the channel impairments, and one would attempt to estimate them as accurately as possible.\",\n \"Modulation Classification: There are a number of well-know techniques for modulation classification.\",\n \"They mostly fall into one of two categories: pattern-recognition based and decision theory-based.\",\n \"The most practical techniques are a hybrid of these two approaches, where a set of key features is extracted from the modulated waveform (as in pattern recognition), and the principles of decision theory are applied to classify the modulation based on those features.\",\n \"Numerous key features have been used for modulation classification.\",\n \"A partial list of those features include: amplitude histograms, frequency histograms, phase histograms, phase difference histograms, the variance of the amplitude, frequency, and phase, higher order moments, kurtosis, cumulants, the square of the signal envelope, zero crossings, the power spectrum of the received signal, the presence of harmonics at selected frequencies, the magnitude of the spectral component at twice the carrier frequency of the signal squared, the magnitude of the spectral component at 4 times the carrier frequency of the signal raised to the fourth power, and the power spectrum asymmetry.\",\n \"If properly chosen and applied, the key features can help discriminate among different modulation formats, even under adverse conditions, such as low signal to noise ratio (SNR), limited amount of data, presence of interference, and channel impairments.\",\n \"Existing modulation classification schemes typically have several shortcomings.\",\n \"One shortcoming is that the key features computation does not take into account that different samples have different reliability values, as they are often taken asynchronously with the signal symbols.\",\n \"Another shortcoming is that the band-limited nature of the waveform (which causes signal fluctuations around the symbol edges) is usually not taken into account A third shortcoming is that the outcome of a classification scheme is often dependent on the sequence of applying the key features.\",\n \"Another shortcoming is that the thresholds used in determining the decision regions are independent of SNR.\",\n \"A further shortcoming is that simple majority rule is used to make a final decision based on the individual segments decisions.\",\n \"Last but not least, is the fact that many existing classification schemes require exact knowledge of the signal parameters, and are not robust to inaccuracies in the value of those parameters.\",\n \"Unfortunately, the schemes that perform the best under idealized conditions tend to be the least robust.\",\n \"Interference Characterization: Interference identification and characterization can significantly enhance the utility of a Communication System Monitoring system.\",\n \"In this regard, “characterization” refers to determining the power level; carrier frequency and occupied bandwidth of the interferer, plus any other transmission parameters that may be estimated.\",\n \"Generalizing the interference characterization to the case of multiple interferers is done iteratively.\",\n \"There are many potential sources of interference in a satellite communication system such as inclined satellites, radars, terrestrial microwave links, in-orbit test equipment generated carriers, rogue transmitters, and carriers on mistaken frequencies and/or directions.\",\n \"In addition, as previously noted, adjacent satellites in the geostationary arc are a main source of interference.\",\n \"Adjacent Satellite Interference (ASI) can occur on the uplink and on the downlink.\",\n \"While the interference mechanism is different in these two cases, both uplink ASI and downlink ASI result in the presence of interfering signals in a frequency band.\",\n \"An interferer's power may be sufficiently low to make its detection and identification difficult, yet sufficiently high to cause noticeable performance degradation to desired signals.\",\n \"Furthermore, the capability to characterize interferers in a desired frequency band can provide useful data on whether other satellite systems are abiding by the frequency coordination agreements to which they are party.\",\n \"A practical algorithm for interference identification and characterization is known in the art.\",\n \"The received waveform consists of a distorted version of the desired signal, thermal noise, and an unknown interferer.\",\n \"It is desired to characterize the interferer to the extent possible.\",\n \"In other words, it is desired to determine the interferer power, center frequency, occupied bandwidth, modulation type, symbol rate, and any other potentially useful information.\",\n \"If one could completely cancel out the desired signal, standard correlation techniques could be used to extract interferer information from the thermal noise.\",\n \"However, the distortion of the desired signal makes its complete cancellation impractical.\",\n \"The goal is then to cancel the desired signal as much as possible so that any residual energy is small and does not mask the presence of an interferer.\",\n \"Impairments that are expected to distort the desired signal waveform include: the non-ideal channel, phase noise, oscillator drift, transmitter non-linearities, non-ideal filtering, clock jitter, intermodulation products, and quadrature imbalance.\",\n \"In order to perform a fairly complete cancellation of the strong signal in this case, understanding the nature and magnitude of the various channel impairments is paramount.\",\n \"Indeed, if by a process of reverse engineering one is able to completely determine all the channel impairments, then one could in principle reconstruct a noise-free, identical copy of the desired signal in the received waveform.\",\n \"Subtracting this constructed replica from the received waveform would leave the interferer and the noise.\",\n \"In reality of course, it is not possible to construct a perfect noise-free copy of the desired signal in the received waveform.\",\n \"The desired approach is to construct as close a replica as possible of the received desired signal by estimating the impairments as accurately as possible.\",\n \"The extent to which it is possible to estimate those impairments and cancel out their effect will determine the degree of success in characterizing the interference.\",\n \"As many of the foregoing processes and procedures are manual or only semi-automated, it is an object of the present invention to provide fully automated procedures for determining each of these satellite performance related parameters.\",\n \"It is also an object of the invention to provide a combination of automated procedures that can attain an automatic generation of satellite frequency plans.\",\n \"It is yet an object of the invention to provide a combination of at least two and possibly all of the automated procedures in order to obtain an optimum result.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is a digital signal processing (DSP)-based approach to parameter estimation, modulation identification and interference characterization in connection with a satellite Communication Monitoring System (CSM).\",\n \"The techniques described here allow automatic generation of satellite frequency plans without any a priori knowledge of such plans.\",\n \"When combined with information publicly available about a given satellite, these techniques will give very precise information of the frequency plan of that satellite.\"\n ],\n [\n \"This application claims the benefit of U.S.\",\n \"Provisional Application Ser.\",\n \"No.\",\n \"60/289,389, filed on May 8, 2002.FIELD OF THE INVENTION The invention relates generally to a satellite communication monitoring (CSM) method and apparatus for providing parameter estimation, modulation classification, and interference characterization in communication satellite systems.\",\n \"BACKGROUND In a satellite communication system, particularly a system where the satellite is deployed in a geostationary orbit, the satellite will be able to receive signals transmitted to the satellite by earth stations at an allocated uplink frequency band and will be operative to transmit signals to earth stations on allocated downlink frequency bands.\",\n \"The uplink bands are selected to be spaced apart from the downlink bands in order to avoid interference.\",\n \"Nonetheless, interference may be generated due to transmissions from adjacent earth stations or adjacent satellites having overlapping beams.\",\n \"In addition, interference may arise from natural phenomena, such as rainfall, scattering, terrestrial communications and the like.\",\n \"With respect to the downlink, the signal received at an earth station from the satellite is frequency-down-converted and digitized by means of an analog-to-digital (A/D) converter.\",\n \"A typical value for the bandwidth of the A/D converted frequency band is 36 MHz.\",\n \"The A/D converter output is a stream of bits which essentially captures all the information in the received signal.\",\n \"FIG.\",\n \"1 illustrates a satellite system 10 having a plurality of satellites 11, 12, 13, 14 in geostationary orbit, including a satellite 13 that is intended to communicate with an earth station 15 and several adjacent satellites 11, 12 and 14.Satellite 13 transmits signals on a downlink band to the earth station 15 having a frequency down converter, A/D converter and CSM equipment 18 that can provide conventional CSM functions.\",\n \"The CSM 18 has an adequate processing capability, which may be provided by a conventional processor (not shown) with appropriate software modules.\",\n \"The CSM system 18 includes a digital spectrum analyzer, operating on the A/D output bits, that provides a panoramic look of the carriers in the entire frequency band that was digitized.\",\n \"In order to properly monitor the received signal, the carriers must be separated or isolated, and subdivided as needed.\",\n \"Then, an estimation made of their parameters, identifying their modulation types, as well as detecting and characterizing any interferer that may be present in the digitized frequency band.\",\n \"The successive steps involve carrier isolation, segmentation, frequency estimation, symbol rate estimation, bit error rate estimation, modulation classification and interference characterization.\",\n \"Carrier Isolation: Carrier isolation consists of identifying and separating the individual carriers in the digitized frequency band.\",\n \"After carrier isolation has been performed, each carrier is processed separately to estimate its parameters, determine its modulation type, etc.\",\n \"Typically, the carriers are identified on a spectrum analyzer by a human operator, in a straightforward and well known process.\",\n \"However, automated carrier detection is difficult, as the process must be capable of differentiating true carriers from thermal noise, statistical fluctuations, side lobes, intermodulation products, and spurious spikes.\",\n \"Segmentation: For the sake of computational simplicity, it is often necessary to segment the time domain data record containing the digital samples into segments of appropriate size, and to process each segment separately.\",\n \"Furthermore, when the channel is not constant, segmentation has the additional advantage of providing a channel which is approximately constant over each segment.\",\n \"Examples of non-constant channels include bursty channels, fading channels, and voice activated channels.\",\n \"The size of the segment is usually chosen as a power of 2 because such a choice leads to the use of efficient FFT processing.\",\n \"FFT processing is the backbone of the digital spectrum analysis to be performed on such segments.\",\n \"Frequency Estimation: There are several well-known techniques to carrier frequency estimation.\",\n \"One popular technique is the centroid method.\",\n \"In this method, the center frequency is estimated as a weighted average frequency, where the weights are taken as the squares of the spectral coefficients.\",\n \"A second method consists of fitting a straight line to the instantaneous phase data.\",\n \"Finding the best straight-line fit is a simple mean square error minimization problem, where the slope of the line provides the frequency estimate and the value at the origin provides the initial phase.\",\n \"When using this technique, the phase values must be unwrapped before the straight line fit A third method consists of passing the received waveform through a nonlinearity, such as quadrupling, and detecting spectral lines at harmonics of the carrier.\",\n \"The frequency location of these spectral lines, which are obtained via a high resolution FET, would provide an accurate estimate of the carrier frequency.\",\n \"While the above three methods are suitable in many situations, they each have their shortcomings, making them unsuitable for some applications.\",\n \"For example, the centroid method is not suitable if the frequency spectrum is not symmetric.\",\n \"The instantaneous phase square error minimization is best suited to constant envelope modulations, and the nonlinearity does not always produce line spectra at harmonics of the carrier frequency.\",\n \"Furthermore, the accuracy provided by these methods may sometimes be insufficient.\",\n \"Symbol Rate Estimation: There are several well-known techniques for symbol rate estimation.\",\n \"One conventional scheme is the delay and multiply method, where the received waveform is multiplied by a replica of itself, that has been delayed by a fraction of the symbol rate.\",\n \"Spectral lines will then appear in the spectrum at harmonics of the symbol rate when the delay is properly chosen.\",\n \"The amount of delay, and the number and magnitude of spectral lines are modulation scheme-dependent and well known in each case.\",\n \"Those spectral lines therefore provide a signature identifying the symbol rate, and may also be used for modulation discrimination.\",\n \"Another method is to use a first order phase lock loop (PLL) to track the timing of the received signal.\",\n \"This is a typical way of achieving clock synchronization in digital modems.\",\n \"While the above methods are suitable for many situations, they each have their shortcomings, making them unsuitable for some applications.\",\n \"For example, the delay and multiply method does not always produce spectral lines at harmonics of the symbol rate.\",\n \"As to the PLL tracking method, it needs a sufficiently accurate knowledge of the symbol rate at the start.\",\n \"BER Estimation: When a signal is demodulated and FEC is decoded, it is possible to obtain an accurate estimate of the BER, without having access to the actual transmitted bits.\",\n \"BER is determined by a well-known procedure based on re-encoding the decoded bits.\",\n \"In the absence of FEC decoding, an accurate BER estimate (coded or uncoded) may be obtained over an AWGN (additive white gaussian noise) channel from accurate estimation of energy per bit/noise density (Eb/No), and knowledge of the modulation format and FEC type and rate.\",\n \"Estimating the uncoded and coded BER becomes more difficult if the channel is not AWGN.\",\n \"In order to provide a fairly accurate BER estimate in this case, understanding the nature and magnitude of the various channel impairments is paramount.\",\n \"Indeed, if by a process of reverse engineering one is able to completely determine all the channel impairments, then one could in principle reconstruct a waveform statistically identical to the one under examination, and therefore one would be able to accurately estimate the BER.\",\n \"In reality of course, it is not possible to completely determine all the channel impairments, and one would attempt to estimate them as accurately as possible.\",\n \"Modulation Classification: There are a number of well-know techniques for modulation classification.\",\n \"They mostly fall into one of two categories: pattern-recognition based and decision theory-based.\",\n \"The most practical techniques are a hybrid of these two approaches, where a set of key features is extracted from the modulated waveform (as in pattern recognition), and the principles of decision theory are applied to classify the modulation based on those features.\",\n \"Numerous key features have been used for modulation classification.\",\n \"A partial list of those features include: amplitude histograms, frequency histograms, phase histograms, phase difference histograms, the variance of the amplitude, frequency, and phase, higher order moments, kurtosis, cumulants, the square of the signal envelope, zero crossings, the power spectrum of the received signal, the presence of harmonics at selected frequencies, the magnitude of the spectral component at twice the carrier frequency of the signal squared, the magnitude of the spectral component at 4 times the carrier frequency of the signal raised to the fourth power, and the power spectrum asymmetry.\",\n \"If properly chosen and applied, the key features can help discriminate among different modulation formats, even under adverse conditions, such as low signal to noise ratio (SNR), limited amount of data, presence of interference, and channel impairments.\",\n \"Existing modulation classification schemes typically have several shortcomings.\",\n \"One shortcoming is that the key features computation does not take into account that different samples have different reliability values, as they are often taken asynchronously with the signal symbols.\",\n \"Another shortcoming is that the band-limited nature of the waveform (which causes signal fluctuations around the symbol edges) is usually not taken into account A third shortcoming is that the outcome of a classification scheme is often dependent on the sequence of applying the key features.\",\n \"Another shortcoming is that the thresholds used in determining the decision regions are independent of SNR.\",\n \"A further shortcoming is that simple majority rule is used to make a final decision based on the individual segments decisions.\",\n \"Last but not least, is the fact that many existing classification schemes require exact knowledge of the signal parameters, and are not robust to inaccuracies in the value of those parameters.\",\n \"Unfortunately, the schemes that perform the best under idealized conditions tend to be the least robust.\",\n \"Interference Characterization: Interference identification and characterization can significantly enhance the utility of a Communication System Monitoring system.\",\n \"In this regard, “characterization” refers to determining the power level; carrier frequency and occupied bandwidth of the interferer, plus any other transmission parameters that may be estimated.\",\n \"Generalizing the interference characterization to the case of multiple interferers is done iteratively.\",\n \"There are many potential sources of interference in a satellite communication system such as inclined satellites, radars, terrestrial microwave links, in-orbit test equipment generated carriers, rogue transmitters, and carriers on mistaken frequencies and/or directions.\",\n \"In addition, as previously noted, adjacent satellites in the geostationary arc are a main source of interference.\",\n \"Adjacent Satellite Interference (ASI) can occur on the uplink and on the downlink.\",\n \"While the interference mechanism is different in these two cases, both uplink ASI and downlink ASI result in the presence of interfering signals in a frequency band.\",\n \"An interferer's power may be sufficiently low to make its detection and identification difficult, yet sufficiently high to cause noticeable performance degradation to desired signals.\",\n \"Furthermore, the capability to characterize interferers in a desired frequency band can provide useful data on whether other satellite systems are abiding by the frequency coordination agreements to which they are party.\",\n \"A practical algorithm for interference identification and characterization is known in the art.\",\n \"The received waveform consists of a distorted version of the desired signal, thermal noise, and an unknown interferer.\",\n \"It is desired to characterize the interferer to the extent possible.\",\n \"In other words, it is desired to determine the interferer power, center frequency, occupied bandwidth, modulation type, symbol rate, and any other potentially useful information.\",\n \"If one could completely cancel out the desired signal, standard correlation techniques could be used to extract interferer information from the thermal noise.\",\n \"However, the distortion of the desired signal makes its complete cancellation impractical.\",\n \"The goal is then to cancel the desired signal as much as possible so that any residual energy is small and does not mask the presence of an interferer.\",\n \"Impairments that are expected to distort the desired signal waveform include: the non-ideal channel, phase noise, oscillator drift, transmitter non-linearities, non-ideal filtering, clock jitter, intermodulation products, and quadrature imbalance.\",\n \"In order to perform a fairly complete cancellation of the strong signal in this case, understanding the nature and magnitude of the various channel impairments is paramount.\",\n \"Indeed, if by a process of reverse engineering one is able to completely determine all the channel impairments, then one could in principle reconstruct a noise-free, identical copy of the desired signal in the received waveform.\",\n \"Subtracting this constructed replica from the received waveform would leave the interferer and the noise.\",\n \"In reality of course, it is not possible to construct a perfect noise-free copy of the desired signal in the received waveform.\",\n \"The desired approach is to construct as close a replica as possible of the received desired signal by estimating the impairments as accurately as possible.\",\n \"The extent to which it is possible to estimate those impairments and cancel out their effect will determine the degree of success in characterizing the interference.\",\n \"As many of the foregoing processes and procedures are manual or only semi-automated, it is an object of the present invention to provide fully automated procedures for determining each of these satellite performance related parameters.\",\n \"It is also an object of the invention to provide a combination of automated procedures that can attain an automatic generation of satellite frequency plans.\",\n \"It is yet an object of the invention to provide a combination of at least two and possibly all of the automated procedures in order to obtain an optimum result.\",\n \"SUMMARY OF THE INVENTION The present invention is a digital signal processing (DSP)-based approach to parameter estimation, modulation identification and interference characterization in connection with a satellite Communication Monitoring System (CSM).\",\n \"The techniques described here allow automatic generation of satellite frequency plans without any a priori knowledge of such plans.\",\n \"When combined with information publicly available about a given satellite, these techniques will give very precise information of the frequency plan of that satellite.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is a schematic illustration of a satellite system where CSM would be utilized.\",\n \"FIG.\",\n \"2 is a flowchart for a procedure that performs an automated carrier isolation process.\",\n \"FIG.\",\n \"3 is a flowchart for a procedure that performs an automated carrier segmentation process.\",\n \"FIG.\",\n \"4 is a flowchart for a procedure that performs an automated carrier frequency estimation process.\",\n \"FIG.\",\n \"5 is a flowchart for a procedure that performs an automated symbol rate estimation process.\",\n \"FIG.\",\n \"6 is a flowchart for a procedure that performs an automated bit error rate (BER) estimation process.\",\n \"FIG.\",\n \"7 is a flowchart for a procedure that performs an automated modulation classification process.\",\n \"FIG.\",\n \"8 is a flowchart for a procedure that performs an automated interference identification and characterization process.\",\n \"FIG.\",\n \"9A is a flowchart for a procedure that integrates three of the foregoing automated processes to achieve an automatic generation of a satellite frequency plan.\",\n \"FIG.\",\n \"9B is a flowchart for a procedure that integrates all of the foregoing automated processes.\",\n \"DETAILED DESCRIPTION OF THE INVENTION In a satellite communication system as illustrated in FIG.\",\n \"1, where a CSM system is available to perform various system monitoring, analysis and estimation functions, the processor resident at the CSM may be operative to perform a variety of procedures, consistent with the algorithms identified subsequently in flowchart form, to automatically estimate parameters, classify modulation and characterize interference in the system.\",\n \"While the invention is disclosed in connection with various specific embodiments, it is not limited thereto and a wide variety of alternative approaches may be evident to one skilled in the art upon reading the following disclosure.\",\n \"For example, the processing performed may be distributed or centralized, with communication with an earth station provided by well known network and system arrangements.\",\n \"Turning now to the individual elements of the signal processing performed in a CSM system contemplated by the present invention, the following procedures may be employed.\",\n \"Carrier Isolation: Carrier isolation would be automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.\",\n \"2.In a first step S1, the processor serving the CSM would perform an FFT of size NFFT on the composite waveform of bandwidth W. The value of NFFT is programmable, and may be given a default value, such as but not limited to 1024.In step S2, the power spectrum is obtained from the FFT output by computing the squared magnitude of each FFT coefficient.\",\n \"This procedure is repeated M times according to step S3 and averaged out in step S4 to smooth out the statistical fluctuations in the spectrum.\",\n \"The value of M is programmable, and in an exemplary embodiment, may have a default value of 16, although other values would be readily apparent to one skilled in the art.\",\n \"Then, a noise floor pn is determined in step S5.With these basic parameters in hand, the power spectrum is filtered in step S6 in order to mitigate the impact of any statistical fluctuations, and to gloss over spurious spikes and frequency nulls between sidelobes.\",\n \"Then, in step S7, a minimum carrier level pc X dB above the noise floor pn is set, where X is a programmable parameter, which may have a default value of 3 dB, or other value as would be apparent to one skilled in the art.\",\n \"The processing will proceed through the individual frequency points from the lowest to the highest, in step S8.When a value higher than pc is first detected at a certain frequency, that frequency is taken as the lower frequency limit of a carrier in step S8.Also, in that same step, when the value at the filter output drops first to a level below pc, the corresponding frequency is taken as the upper frequency limit of the carrier.\",\n \"The entire spectrum is processed according to the procedure of step S8, as indicated in step S9, thus identifying the lower and upper frequency limits for each carrier.\",\n \"Finally, in step S10, the individual carriers are digitally filtered out, one by one, in accordance with the procedure identified above.\",\n \"Thereafter, the procedure comes to an end.\",\n \"Segmentation: Segmentation would be automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.\",\n \"3.In a first step S11, a number of samples that are to be included in a segment are determined and a suitable power of 2 is identified to represent the selected number of samples in the segment.\",\n \"Then, in step S12, the instantaneous power in each sample is computed.\",\n \"Next, in step S13, the instantaneous power values are filtered out in order to remove large deviations.\",\n \"In step S14, the average and the standard deviation of the instantaneous power is computed and, in step S15, the normalized standard deviation is computed and compared to a threshold based on the value of Eb/No, where Eb is the signal strength and No is a corresponding noise value.\",\n \"In step S16, it is determined whether the threshold is exceeded.\",\n \"If not exceeded, the segment is rejected (N), otherwise (Y), it is accepted in step S17.Then in step S18, it is determined whether the analyzed segment is the last segment and, if not, the process proceeds to test the next segment If it is the last segment, the process ends.\",\n \"Frequency Estimation: Frequency estimation is automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.\",\n \"4.In a first step S21, the processor would compute a frequency estimate using the well known centroid method.\",\n \"Then, in step S22, the instantaneous phase method is modified.\",\n \"The modification would be made as follows.\",\n \"Assign a weight proportional to the square of the magnitude to each instantaneous phase value, so that samples with higher SNR are given more weight.\",\n \"Use a weighted minimum mean square error criterion.\",\n \"Then, in step S23, a second frequency estimate is computed using the modified instantaneous phase method as described above.\",\n \"In step S24, the waveform is passed through a non-linearity and any harmonics in the spectrum are detected.\",\n \"In step S25, the FFT-based location of harmonics is enhanced by using unbiased interpolation of the FFT coefficients.\",\n \"Finally, in step S26, a third frequency estimate is determined on the basis of the enhanced harmonics location process, as previously disclosed.\",\n \"Once the three frequency estimates are obtained, although more may be obtained if desired, a weighted average of the frequency estimates is determined in step S27.The weights are assigned on the basis of spectral symmetry, envelope fluctuations, and strength of the frequency harmonics.\",\n \"As would be understood by one skilled in the art, if only a moderate frequency estimation accuracy is sought, a subset of the above set of estimates would be adequate.\",\n \"On the other hand, if higher frequency accuracy is still needed, supplement the estimate obtained above with a phase locked loop to track the received carrier.\",\n \"Symbol Rate Estimation: Symbol rate estimation is automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.\",\n \"5, using a modification of the conventional delay and multiply technique.\",\n \"According to the modification, in a step S31, both the received signal and its delayed replica are passed through a nonlinearity to produce harmonics at the symbol rate.\",\n \"Then, in step S32, the processor would compute the number of crossings per unit time where the signal envelope crosses the half power level.\",\n \"In step S33, the timing (clock frequency and phase) of the received waveform would be tracked, in an exemplary embodiment, by using a second order PLL.\",\n \"In a subsequent process represented by step S34, a non-uniform sampling approach would be used.\",\n \"For example, but without limitation, a non-uniformly sampled set may be generated by digital interpolation between the available uniformly sampled samples.\",\n \"The proposed non-uniform sampling rate is slowly and monotonically increasing, and covers the range of uncertainty in the symbol rate.\",\n \"This provides the ability to home in on the true sample rate.\",\n \"Once lock is achieved, uniform sampling is resumed and a PLL is used to fine tune the symbol rate estimate.\",\n \"If only a moderate symbol rate estimation accuracy is sought, a subset of the above set of estimates would be adequate.\",\n \"BER Estimation: Bit error rate estimation is automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.\",\n \"6.According to step S41, the process begins with an estimate the waveform parameters and a determine the modulation type, if it is not already known, as described above.\",\n \"Then, in step S42, there is a processing of the received samples with a properly matched and equalized filter, followed by a tracking of the carrier phase and the clock phase in step S43.In step S44, the well known maximum likelihood techniques are used to estimate the phase noise, intermodulation products, quadrature imbalances, and non-linearity's.\",\n \"Any side information available regarding the transmitter characteristics, such as for example the power amplifier specifications, may be used for this purpose in step S45.The information may be available beforehand and either input manually or accessible automatically by the processor on the basis of pre-stored information in RAM or auto detected characteristics of the equipment, in a manner known in the art.\",\n \"The process proceeds in option 1 to the construction of a waveform with the estimated parameters and modulation type, subject it to the estimated impairments, and estimate the BER, in step S46.Alternatively, the process may proceed as option 2 to step S47 by first constructing a noise-free scattering diagram based on the estimated impairments.\",\n \"Then, an estimate of the uncoded and coded BER, using maximum likelihood, may be obtained from the noise-free scattering diagram, and the estimated Eb/No in step S48.Modulation Classification: Modulation classification is automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.\",\n \"7.According to step S51, an estimate is automatically made of the waveform parameters as accurately as possible, as outlined above.\",\n \"Then, an estimate of the signal-to-noise ratio (SNR) of the received waveform is obtained in step S52.Available side information, if any, may be used to narrow down the set of potential modulation formats at this point, according to step S53.The information may be available beforehand and either input manually or accessible automatically by the processor on the basis of pre-stored information in RAM or auto detected characteristics of the equipment, in a manner known in the art.\",\n \"Then several modification steps occur.\",\n \"In step S54, the key features computation is modified such that each sample contributing to a key feature is assigned a weight proportional to its SNR.\",\n \"(Some phase samples are more sensitive to noise than others, depending on the magnitude of those samples.)\",\n \"In step S55, the key features computation is modified such that each sample contributing to a key feature is assigned a weight proportional to its distance from the symbol edges.\",\n \"(Band limiting causes envelope fluctuations around the symbol edges).\",\n \"In step S56, based on the side information, a subset of key features from the set listed above is computed.\",\n \"Then, in step S57, the sub-optimum hierarchical classification approach to a vector approach is modified, where several features are applied simultaneously to a multidimensional threshold.\",\n \"The threshold setting is made SNR-dependent.\",\n \"(Actual threshold values for different SNRs are computed offline).\",\n \"In step S58, the number of segments processed is made SNR-dependent to achieve a given confidence level.\",\n \"And, in step S59, for each segment processed, a ranking is assigned as to how likely it is that the waveform under examination belongs to each of the modulation classes under consideration.\",\n \"Finally, in step S60, a soft combining of all the segment rankings is performed to arrive at the most likely overall classification of a modulation type.\",\n \"Interference Characterization: Interference characterization is automatically performed in accordance with the following procedure, consistent with the flowchart illustrated in FIG.\",\n \"8.According to step S61, an estimate is made of the waveform parameters and a determination is made of the modulation type of the desired signal, if it is not already known, as described earlier.\",\n \"In step S62, the received samples are processed with a properly matched and equalized filter.\",\n \"Then, in step S63, the carrier phase and the clock phase are tracked.\",\n \"Any side information available regarding the transmitter characteristics, such as for example the power amplifier specifications, may be used in this regard and optionally input.\",\n \"The information may be available beforehand and either input manually or accessible automatically by the processor in step S63A on the basis of pre-stored information in RAM or auto detected characteristics of the equipment, in a manner known in the art.\",\n \"In step S64, the well known maximum likelihood techniques is used to estimate the phase noise, intermodulation products, quadrature imbalances, and nonlinearities.\",\n \"Then, in step S65, the received signal is demodulated and the transmitted bits are recovered.\",\n \"Optionally, if the SNR is low and the error rate is high, FEC decoding of the signal to recover the information bits can be beneficial in this step.\",\n \"If FEC decoding was performed, the information bits must be re-encoded.\",\n \"In step S66, the transmitted bits are remodulated on a carrier according to the known (or estimated) modulation type, symbol rate, and filter characteristics.\",\n \"The remodulated signal is subjected to the impairments estimated above in step S67 and the remodulated signal from the received waveform in step S68.A standard correlation and spectral analysis techniques is performed on the residual signal to extract interferer information from the noise, in step S69.The several processes for automated determination of parameters may be combined to provide an automatic generation of a satellite frequency plan, as illustrated in FIG.\",\n \"1, and according to the process of FIG.\",\n \"9A.\",\n \"Specifically, the carrier is isolated automatically, according to the process in FIG.\",\n \"2, in step S71.The carrier isolation is followed by a segmentation processing in step S72, according to the flowchart of FIG.\",\n \"3.Thereafter, a frequency estimation process according to the method of FIG.\",\n \"4 is conducted automatically in step S73.The result of this combination of outputs would be automatically combined into a frequency plan for a satellite by the CSM in step S74.The several processes disclosed in FIGS.\",\n \"2-8 may be conducted automatically, in any combination, as would be known in the art, including a combination of all of the processes as illustrated in FIG.\",\n \"9B.\",\n \"There, as in FIG.\",\n \"9A, the carrier isolation, segmentation and frequency estimation processes, which derive a frequency plan in step 81, may be accompanied by the estimation of symbol rate according to the process of FIG.\",\n \"5 in step S82 and the estimation of BER according to the process of FIG.\",\n \"6 in step S83 The modulation classification according to FIG.\",\n \"7 may be performed in step S84 and the interference characterization according to FIG.\",\n \"8 may be performed in step S85.While the present invention has been described in accordance with certain embodiments and examples, it is not limited thereto.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476869"}}},{"rowIdx":979,"cells":{"text":{"kind":"list like","value":[["Method and apparatus for measuring adjacent satellite interference","A system for determining at least one of uplink ASI and downlink ASI in a satellite communication system having at least a first satellite and a second satellite, each having uplink and downlink footprints and being in communication with respective earth stations and having a potential interference.","The system comprises a first antenna for receiving downlink signals communicated from said first satellite, a second antenna for receiving downlink signals communicated from said second satellite, power spectrum measurement equipment connected to the second antenna for measuring the second satellite power spectrum, and CSM equipment connected to the first antenna and the power spectrum measurement equipment for generating an adjacent satellite carrier plan."],["1.A method of characterizing downlink ASI in a communications satellite system having at least a first satellite and at least a second adjacent satellite, each said satellite being in communication with a respective earth station via downlink beams that provide overlapping downlink beam footprints, said method comprising: a) capturing the power spectrum of the second adjacent satellite; b) dividing the downlink beam of the second adjacent satellite into transponder segments; c) performing spectrum analysis on each said transponder segment; d) analyzing the measured power spectrum of the second adjacent satellite to convert measured transponder spectra to estimates of carrier frequency plans.","2.The method of characterizing downlink ASI as recited in claim 1, further comprising, determining downlink ASI on a system-wide basis by repeating the process for each downlink beam of the first satellite, and for each of plural adjacent satellites.","3.The method characterizing downlink ASI as recited in of claim 1 further comprising using different measurement locations for each of different downlink beams that cover different geographical regions.","4.The method of characterizing downlink ASI as recited in claim 1 further comprising pointing a CSM antenna towards the adjacent satellite.","5.The method of characterizing downlink ASI as recited in claim 1 further comprising using spectral parameters of power, center frequency and spectral shape, and optionally at least one of modulation type, FEC coding and scrambling patterns.","6.A method of characterizing uplink ASI in a communications satellite system having at least a first satellite and a second adjacent satellite, each said satellite being in communication with respective earth stations via downlink beams that provide downlink beam footprints and uplink beams that have uplink beam footprints, said method comprising: a) using the downlink spectrum of the interfering satellite; and b) processing the downlink in accordance with the method of 1) capturing the power spectrum of the second adjacent satellite via spectrum measurement equipment; 2) dividing the downlink beam of the second adjacent satellite into segments; 3) performing spectrum analysis on each said segment; and 4) analyzing the measured power spectrum of the second adjacent satellite to convert measured transponder spectra to estimates of carrier frequency plans.","7.The method of characterizing uplink ASI as claimed in claim 6, further comprising: if the second adjacent satellite uses the same uplink and downlink frequency bands as the first satellite, then using only the power spectrum measurement of the second adjacent satellite's downlink spectrum to represent an uplink ASI.","8.The method of characterizing uplink ASI as claimed in claim 6, further comprising: if either the uplink or the downlink of the second adjacent satellite operates in frequency bands that are different from the first satellite, determining whether the downlink frequency band is the same, and if the uplink of the second adjacent satellite shares the same frequency band with the first satellite, but where the downlink of the second adjacent satellite uses a different band, receiving the downlink of the second adjacent satellite and characterizing uplink ASI.","9.The method of characterizing uplink ASI as claimed in claim 6, further comprising repeating the characterization of uplink ASI on a system-wide basis, by repeating the process for each uplink beam of the first satellite, and for each adjacent satellite.","10.A method of characterizing uplink ASI as recited in claim 6 further comprising using the derived information in identifying the source of interference.","11.A system for determining at least one of uplink ASI and downlink ASI in a satellite communication system having at least a first satellite and a second satellite, each having uplink and downlink footprints and being in communication with respective earth stations and having a potential interference, comprising: a first antenna for receiving downlink signals communicated from said first satellite; a second antenna for receiving downlink signals communicated from said second satellite; power spectrum measurement equipment connected to said second antenna for measuring the second satellite power spectrum; and CSM equipment connected to said first antenna and said power spectrum measurement equipment for generating an adjacent satellite carrier plan.","12.The system as set forth in claim 11 wherein said second antenna is a steerable antenna.","13.The system as set forth in claim 11 further comprising at least one of: a) means for providing ASI interference identification; b) means for providing data for inter-system coordination; c) means for providing verification of intra-system ASI provisions; and d) means for system transmission planning.","14.The system as set forth in claim 13, wherein said means for system transmission planning comprises: means for analysis of ASI effects on a per carrier basis; and means for carrier by carrier power level optimization.","15.The system as set forth in claim 11, further comprising means for determining whether each of the uplink and downlink frequency bands for the first and second satellites are the same and, if the same, using the power spectrum of the downlink to represent uplink ASI."],["<SOH> FIELD OF THE INVENTION <EOH>The invention relates generally to a method and apparatus for measuring adjacent satellite interference (ASI) using communications system monitoring equipment, for both up-link and down-link interference.","ASI is the interference caused to a victim satellite and one or more of the earth stations with which it is in communication by transmissions from an interfering satellite or the earth stations with which it communicates.","Typically, ASI is caused by earth station antenna sidelobes or as a result of antenna mis-pointing.","The characterization of adjacent satellite interference (ASI) is a function that is not normally performed by communications system monitoring (CSM) systems implemented in commercial communications satellite networks.","However, due to decreased satellite spacing along the geostationary arc, which increases the severity of ASI, it has now become appropriate that the transmissions of adjacent satellites should be monitored, in order to characterize ASI.","Such monitoring would permit coordination agreements to be policed, and would allow carrier frequency plans to be optimized to maximize transponder capacity in the presence of ASI.","FIG.","1A illustrates a typical satellite system, comprising plural geostationary satellites 1 , 2 , 3 4 , which include two satellites 3 , 4 with beams having respective footprints 6 and 7 (spacecraft antenna coverage areas) that overlap on the surface of the earth 8 .","An earth station 5 that is located within the two beam footprints 6 , 7 is designed to transmit to satellite 4 , but its signal also is transmitted to adjacent satellite 3 .","Uplink ASI occurs when the earth stations like station 5 are transmitting to an adjacent satellite 4 (in the same frequency band) and cause interference to a victim satellite 3 , either via the earth station's antenna sidelobes, or via the main lobe due to antenna mis-pointing.","Similarly, earth stations transmitting to satellites 1 and 2 may also cause uplink ASI.","FIG.","1B also illustrates a typical satellite system, comprising plural geostationary satellites 11 , 12 , 13 14 , which include two satellites 13 , 14 with beams having respective footprints 16 and 17 that overlap on the surface of the earth 8 .","An earth station 15 that is located within the two beam footprints 16 , 17 is designed to receive from satellite 13 , but it also receives a signal transmitted by adjacent satellite 14 .","Downlink ASI occurs when earth stations like station 15 that receive transmissions from the victim satellite 13 also receive interference from one or more adjacent satellites 14 (and possibly 11 and 12 ), either via the earth station's antenna sidelobes, or as a result of earth station antenna mis-pointing.","In uplink ASI, it is the earth station transmissions of the interfering satellite's network that produce the interference, while for downlink ASI, it is the earth station receiving systems of the victim satellite's network that contribute to the interference.","In both uplink and downlink ASI, ASI can be produced by uplink and downlink emissions that exceed authorized levels, in addition to the earth station imperfections noted above.","In both uplink and downlink ASL it is a combination of closely spaced satellites and the use of small earth station antennas that increases the severity of ASI.","Currently, CSM systems include a capability to monitor (modulation type, FEC coding, scrambling patterns, bit error rate (BER), symbol rate estimation, etc.)","but do not include the use of equipment for power spectrum measurement and analysis of adjacent satellites.","In short, the conventional CSM systems do not provide a capability to monitor ASI, notwithstanding the improved processing capability that new generation CSM systems offer.","Accordingly, it is an object of the present invention to provide a method and apparatus for CSM systems, that permit existing systems to be enhanced at a modest cost to include ASI characterization"],["<SOH> SUMMARY OF THE INVENTION <EOH>The invention involves a method and apparatus for the provision of a CSM enhancement to include ASI characterization.","The invention more particularly involves a method of characterizing downlink ASI in a communications satellite system having at least a first satellite and at least a second adjacent satellite, each satellite being in communication with a respective earth station via downlink beams that provide overlapping downlink beam footprints.","The downlink ASI determination method comprises: a) capturing the power spectrum of the second adjacent satellite; b) dividing the downlink beam of the second adjacent satellite into transponder segments; c) performing spectrum analysis on each said transponder segment; and d) analyzing the measured power spectrum of the second adjacent satellite to convert measured transponder spectra to estimates of carrier frequency plans.","The invention further comprises a method of characterizing uplink ASI in a communications satellite system having at least a first satellite and a second adjacent satellite, each satellite being in communication with respective earth stations via downlink beams that provide downlink beam footprints and uplink beams that have uplink beam footprints.","The method comprises a) using the downlink spectrum of the interfering satellite; and b) processing the downlink in accordance with the above method.","The invention also is directed to a system for determining at least one of uplink ASI and downlink ASI in a satellite communication system having at least a first satellite and a second satellite, each having uplink and downlink footprints and being in communication with respective earth stations and having a potential interference.","The system comprises a first antenna for receiving downlink signals communicated from said first satellite, a second antenna for receiving downlink signals communicated from said second satellite, power spectrum measurement equipment connected to the second antenna for measuring the second satellite power spectrum, and CSM equipment connected to the first antenna and the power spectrum measurement equipment for generating an adjacent satellite carrier plan."],["This application claims the benefit of U.S.","Provisional Application Ser.","No.","60/289389, filed on May 8, 2002.FIELD OF THE INVENTION The invention relates generally to a method and apparatus for measuring adjacent satellite interference (ASI) using communications system monitoring equipment, for both up-link and down-link interference.","ASI is the interference caused to a victim satellite and one or more of the earth stations with which it is in communication by transmissions from an interfering satellite or the earth stations with which it communicates.","Typically, ASI is caused by earth station antenna sidelobes or as a result of antenna mis-pointing.","The characterization of adjacent satellite interference (ASI) is a function that is not normally performed by communications system monitoring (CSM) systems implemented in commercial communications satellite networks.","However, due to decreased satellite spacing along the geostationary arc, which increases the severity of ASI, it has now become appropriate that the transmissions of adjacent satellites should be monitored, in order to characterize ASI.","Such monitoring would permit coordination agreements to be policed, and would allow carrier frequency plans to be optimized to maximize transponder capacity in the presence of ASI.","FIG.","1A illustrates a typical satellite system, comprising plural geostationary satellites 1, 2, 3 4, which include two satellites 3, 4 with beams having respective footprints 6 and 7 (spacecraft antenna coverage areas) that overlap on the surface of the earth 8.An earth station 5 that is located within the two beam footprints 6, 7 is designed to transmit to satellite 4, but its signal also is transmitted to adjacent satellite 3.Uplink ASI occurs when the earth stations like station 5 are transmitting to an adjacent satellite 4 (in the same frequency band) and cause interference to a victim satellite 3, either via the earth station's antenna sidelobes, or via the main lobe due to antenna mis-pointing.","Similarly, earth stations transmitting to satellites 1 and 2 may also cause uplink ASI.","FIG.","1B also illustrates a typical satellite system, comprising plural geostationary satellites 11, 12, 13 14, which include two satellites 13, 14 with beams having respective footprints 16 and 17 that overlap on the surface of the earth 8.An earth station 15 that is located within the two beam footprints 16, 17 is designed to receive from satellite 13, but it also receives a signal transmitted by adjacent satellite 14.Downlink ASI occurs when earth stations like station 15 that receive transmissions from the victim satellite 13 also receive interference from one or more adjacent satellites 14 (and possibly 11 and 12), either via the earth station's antenna sidelobes, or as a result of earth station antenna mis-pointing.","In uplink ASI, it is the earth station transmissions of the interfering satellite's network that produce the interference, while for downlink ASI, it is the earth station receiving systems of the victim satellite's network that contribute to the interference.","In both uplink and downlink ASI, ASI can be produced by uplink and downlink emissions that exceed authorized levels, in addition to the earth station imperfections noted above.","In both uplink and downlink ASL it is a combination of closely spaced satellites and the use of small earth station antennas that increases the severity of ASI.","Currently, CSM systems include a capability to monitor (modulation type, FEC coding, scrambling patterns, bit error rate (BER), symbol rate estimation, etc.)","but do not include the use of equipment for power spectrum measurement and analysis of adjacent satellites.","In short, the conventional CSM systems do not provide a capability to monitor ASI, notwithstanding the improved processing capability that new generation CSM systems offer.","Accordingly, it is an object of the present invention to provide a method and apparatus for CSM systems, that permit existing systems to be enhanced at a modest cost to include ASI characterization SUMMARY OF THE INVENTION The invention involves a method and apparatus for the provision of a CSM enhancement to include ASI characterization.","The invention more particularly involves a method of characterizing downlink ASI in a communications satellite system having at least a first satellite and at least a second adjacent satellite, each satellite being in communication with a respective earth station via downlink beams that provide overlapping downlink beam footprints.","The downlink ASI determination method comprises: a) capturing the power spectrum of the second adjacent satellite; b) dividing the downlink beam of the second adjacent satellite into transponder segments; c) performing spectrum analysis on each said transponder segment; and d) analyzing the measured power spectrum of the second adjacent satellite to convert measured transponder spectra to estimates of carrier frequency plans.","The invention further comprises a method of characterizing uplink ASI in a communications satellite system having at least a first satellite and a second adjacent satellite, each satellite being in communication with respective earth stations via downlink beams that provide downlink beam footprints and uplink beams that have uplink beam footprints.","The method comprises a) using the downlink spectrum of the interfering satellite; and b) processing the downlink in accordance with the above method.","The invention also is directed to a system for determining at least one of uplink ASI and downlink ASI in a satellite communication system having at least a first satellite and a second satellite, each having uplink and downlink footprints and being in communication with respective earth stations and having a potential interference.","The system comprises a first antenna for receiving downlink signals communicated from said first satellite, a second antenna for receiving downlink signals communicated from said second satellite, power spectrum measurement equipment connected to the second antenna for measuring the second satellite power spectrum, and CSM equipment connected to the first antenna and the power spectrum measurement equipment for generating an adjacent satellite carrier plan.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1A is a schematic illustration of a satellite system in which Uplink ASI occurs.","FIG.","1B is a schematic illustration of a satellite system in which Uplink ASI occurs.","FIG.","2 is a schematic illustration of a satellite system having a CSM capability that includes ASI detection.","FIG.","3A is a flow chart that identifies a process for determining downlink ASI.","FIG.","3B is a flow chart that identifies a process for determining uplink ASI.","DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG.","2 illustrates the ASI-related features of a satellite system 20 having a plurality of satellites 21, 22, 23, 24 in geostationary orbit, including a satellite that is desired for communication 23 and an adjacent interfering satellite 24.The desired satellite 23 is in communication with an earth station 25 having CSM equipment 28 that can provide conventional CSM functions.","Those conventional functions are supplemented by equipment That can determine an adjacent satellite carrier plan.","Specifically, a second antenna 26, which may be a shared and steerable antenna that serves several satellites, or a separate dedicated antenna, is provided with equipment that can measure the adjacent satellite 24 power spectrum.","That power spectrum output is provided to the CSM 28, which also has the capability 29 to identify the adjacent satellite carrier plan.","It also has the capability to characterize ASI for several purposes, including identification of ASI interference 30, provide data for inter-system coordination 31, verify intra-system ASI provisions 32 and utilize system transmission planning software 33 for a variety of purposes.","Most importantly, the software can be used to provide an analysis of ASI effects on a per carrier basis 34 and provide carrier by carrier power level optimization 35 in order to ensure maximum capacity in the presence of ASI.","The CSM equipment 28 also must have an adequate processing capability, provided by a conventional processor (not shown) with appropriate software modules.","Finally, there must be a means for conveying adjacent satellite spectral measurements to the location where ASI processing is performed.","Depending on the network architecture employed by the basic CSM system, and the extent to which the required processing functions are centralized, additional provisions may be incorporated to convey ASI power spectrum measurements to the locations where ASI information is further processed and used.","While the design of the processing and distribution architecture involves tradeoffs between the extent of local processing that is performed, and the amount of data required to be transported, these considerations would not be major issues for a CSM enhancement, particularly if ASI power spectrum measurements are made infrequently.","The antenna 26 that is used to measure emissions from adjacent satellites not only should be able to be pointed along the geostationary arc, but should also be capable of receiving both linear and circular polarizations of each sense, and over bands that may extend beyond the victim satellite's downlink band.","If downlink ASI were only of interest, then only the polarization sense(s) and bandwidth of the victim satellite's downlinks need be measured.","However if uplink ASI characterization is required, other polarizations and other receive bands may be required to obtain the required replicas of the interfering satellite's uplink spectrum.","This requirement is due to the fact that the interfering satellite may use the same uplink bands as the victim satellite, but different downlink bands.","Turning next to the application of the system of FIG.","2 to the solution of practical problems, is should be noted first that there are differences in uplink and downlink ASI, as further detailed subsequently.","However, since the implementation to characterize downlink ASI is somewhat simpler than for uplink ASI, downlink ASI is described first.","In downlink ASI, the footprints of both the victim and the interfering satellite overlap in the geographical areas that experience ASI.","Also, the downlink spectrum used by affected transponders of the victim satellite overlaps that of the interfering satellite.","The objective of downlink ASI characterization is to obtain a description of the interfering satellite's downlink carrier plan in the transponders of the interfering satellite that overlap both the coverage area and frequency band of the victim satellite.","Downlink ASI characterization can be used to assist in identifying the source of interference to the victim satellite 23, in the inter-system coordination process with the operator of the adjacent satellite 24, and to verify that the adjacent satellite's frequency plan matches detailed ASI database information (for cases where the adjacent satellite 24 has the same operator as the victim satellite 23).","As indicated in function 34 of FIG.","2, an important use of the adjacent satellite's carrier frequency plan is to incorporate it into the transmission analysis and planning functions of computing the effects of downlink ASI on a carrier-by-carrier basis.","A further function, as indicated in function 35 of FIG.","2 is optimizing the carrier frequency plan of the victim satellite to ensure maximum throughput in the presence of ASI.","This latter application is useful in cases where either by agreement, or by violation of coordination agreements, ASI is significant enough to require that transmission planning be performed on a carrier-by-carrier basis, rather than for entire bandwidth segments.","The procedure to characterize downlink ASI is illustrated in the flowchart of FIG.","3A.","A first step S1 is to point the CSM antenna 26 (or equivalently, connect to another dedicated antenna whose output can be processed by the CSM 28) towards the adjacent satellite.","A second step S2 is to “capture” the power spectrum of the adjacent satellite 24 via the spectrum measurement equipment 27.A third and fourth step S3 and S4 involve the performance of analysis functions in the CSM.","For this purpose, spectrum analysis should be done in segments that match the transponder frequency ranges of the victim satellite 23.Accordingly, in step S3, the downlink beam is divided into segments and in step S4 the spectrum analysis is performed on the segment.","This measurement must be repeated for all segments until all segments are analyzed.","Thus, in step S5, a determination is made as to whether the last segment has been analyzed, and if not, the process loops back to step S4.If it is the last segment, then every transponder segment transmitted by the downlink beam of the interfering satellite 24, which was measured in steps S1 and S2, that causes ASI to one or more downlink beams of the victim satellite 23 has been tested.","To characterize downlink ASI on a system wide basis, the process illustrated by FIG.","3A is repeated for each downlink beam of the desired satellite 23, and for each of the adjacent satellites (21, 22, and 24).","Note that to characterize downlink ASI for different downlink beams of the desired satellite 23, different measurement locations are required for each of the downlink beams since they cover different geographical regions (except for the case of polarization reuse beams, where in some cases, one measurement site may be able to be used for two beams).","The measured power spectrum of the adjacent satellite 24 is analyzed in step S6 with software that converts measured transponder spectra to estimates of carrier frequency plans 29.Then, the program ends at step S7.Although for the purposes of ASI interference computation, only the spectral parameters of power, center frequency and spectral shape are required, other parameters such as modulation type, the use of FEC coding, scrambling patterns, etc., that may be outputs of the basic CSM spectral analysis software, also may be used in documenting and coordinating ASI.","Although the way in which information on uplink ASI is used is similar to the way information on downlink ASI is used, the uplink ASI generation mechanism is different from downlink ASI, as earlier noted.","In uplink ASI, it is earth stations transmitting to an adjacent satellite 24 that are the source of interference.","Assuming that the adjacent satellites 24 are of the conventional bent-pipe type (i.e., that the adjacent satellites do not incorporate on-board processing) the way to characterize uplink ASI is to monitor the downlink of the transponder, which has the carriers that generate uplink ASI, in the adjacent satellite 24.Since it is often the case in commercial satellites that the transponder uplink beam coverages do not overlap the downlink beam coverages, the earth stations that measure the spectrum of the adjacent satellite's transponder, whose carriers are the source of uplink ASI, often must be located far from the uplink coverage area where the uplink ASI is produced.","Thus, while to characterize downlink ASI, little information is required about the adjacent satellite except for its location and downlink footprint (i.e., all other information relevant to downlink ASI can be measured) more information about the interfering satellite is required to characterize uplink ASI.","Specifically, both the uplink and downlink beam footprints, the uplink and downlink frequency bands, and the connectivity between the uplink and the transponders of the interfering satellite must be known, so that the locations of the monitoring earth stations can be determined.","Thus, in a flowchart in FIG.","3B representing the uplink ASI characterization process, the first step S11 is to determine station location information In step S12, it is then determined if the adjacent satellite 24 uses the same uplink and downlink frequency bands as the victim satellite 23.If so, then every power spectrum measurement of an adjacent satellite's downlink spectrum (on a transponder-by-transponder basis) will represent both an uplink ASI and a downlink ASI spectrum, and such measurement would be used in step S13.However, this is not the case if either the uplink or the downlink of the adjacent satellite 24 operates in frequency bands that are different from the victim satellite 23.A determination is made in step S14 of whether the downlink frequency band is the same.","For those cases where the uplink of the interfering satellite 24 shares the same frequency band, but where the downlink of the interfering satellite 24 uses a different band (such as, for example, in the case of a cross-strapped transponder) the downlink of the interfering satellite 24 must be received in order to characterize uplink ASI, even though the downlink is in a band that is different from the downlink bands used by the victim satellite 23.To characterize uplink ASI on a system-wide basis, the process illustrated by FIG.","3B is repeated for each uplink beam of the desired satellite, and for each adjacent satellite.","Once the uplink ASI power spectrum is measured, its characterization and its use are essentially the same as previously described for downlink ASI.","That is, information on uplink ASI can be used to assist in identifying the source of interference, as in function 30.It also can assist is coordinating ASI problems with the operator of the adjacent satellite, as in function 31.A further function 32 is to verify that the adjacent satellite's carrier plan conforms to ASI database information in the case where the adjacent satellite is under control of the same operator.","Also, uplink ASI can be used with software 33 to compute the effects of ASI on a carrier-by-carrier basis, as in function 34 and optimize capacity in transponders subject to significant levels of ASI, as in function 35.A transmission planning and analysis program 33 would be implemented in a manner known in the art.","The software required for ASI characterization is in two basic categories.","First, there is the software required to convert a measured power spectrum into an estimated ASI carrier frequency plan.","Second, there is the software that uses the ASI characterization results.","It is only the former category that requires processing by the basic CSM equipment, and this processing requirement should be modest since the computations need not be made in real time, and, as noted above, ASI power spectrum measurements should only be required relatively infrequently.","The software required for the purposes of using the ASI characterization results is part of the system operations, planning, and inter-system coordination functions.","Hence, any hardware associated with these software functions would not be part of the basic CSM system.","Returning to the first category of software required for ASI characterization, namely that required to convert the measured adjacent satellite's power spectrum into an estimate of the adjacent satellite's carrier frequency plan (over the portion of bandwidth of interest), there are two subcategories.","A first subcategory is software that performs this function for the transponders of the desired satellite 23, as part of a basic CSM system.","The requirements for this software are not discussed herein as they are known.","However, the second subcategory involves software that defines what ASI measurements are required to be made.","This software would use information on adjacent satellites 24 to define the direction, frequency bands, and polarization that are required for ASI measurements.","The complexity of the software required for this subcategory is minimal, and would be easily acquired by one skilled in the art, once the requirement is known.","If only downlink ASI were being characterized, an alternative “search mode” procedure could be used to search for satellites along the geostationary arc having emissions in the same band and polarization as the downlink of the desired satellite.","Assuming that the footprints of the adjacent satellites overlap with the desired satellite, a procedure for downlink characterization would not require the subcategory of measurement control software identified above.","Turning now to the second category of software, it includes software that makes use of the estimated adjacent satellite carrier plans.","Applications in this category include: a) Software that assists users in identifying interference measured by the CSM for the desired satellite as having ASI, and if so, whether it is uplink or downlink ASI.","b) Software that generates adjacent satellite carrier plans in a format useful for intersystem coordination.","c) Software that compares a measured adjacent satellite carrier plan against ASI information stored in a database.","(Useful for system operators that have multiple adjacent satellites.)","d) Transmission planning and analysis software that computes the effects of ASI on the performance of carriers, and optimizes carrier plans to maximize capacity in the presence of ASI.","An example of the last category of software would be the well known STRIP7 and COMPLAN programs developed by LMGT (formerly COMSAT Laboratories).","The STRIP7 program is specialized to Intelsat's requirements, and has a provision for computing impairments caused by intra-system ASI from its own satellites.","The COMPLAN program, which is applicable to non-Intelsat satellites, requires a straightforward modification to handle ASI impairments.","These two programs (or software with similar capabilities) perform two important functions for an FDMA (frequency division multiple access) system.","The first function is that they analyze the performance of every carrier in a satellite transponder, taking into account impairments due to thermal noise, intermodulation noise, intra-satellite co-channel interference, adjacent carrier interference, and uplink and downlink rain impairments.","To these impairments, ASI must be added in the software envisioned for this application (which is already implemented in STRIP7, as noted above).","Since the amount of ASI is dependent on both the characteristics of the uplink earth stations of the interfering satellite, and the downlink earth stations of the victim satellite, the ability to analyze transmission performance on an individual carrier basis is important, since such performance estimates cannot be accurately made solely from the carrier plan of the adjacent satellite.","The second function of these two transmission analysis programs is that given the carrier frequency assignments of a frequency plan, the uplink carrier powers are optimized so that the power-limited capacity of the transponder is maximized.","This is done by computing the effects of the impairments noted above on each individual carrier and providing each carrier with the amount of power it requires to achieve specified performance, while maintaining an optimum transponder operating point (i.e., while maintaining an optimum balance between downlink thermal noise, intermodulation noise, and interference.)","The utility of applying ASI to transmission planning and analysis software is that for situations where ASI is significant, and cannot be eliminated by either intra- or inter-system coordination, the effects of ASI can be quantified on an individual carrier basis, and the transmission plan of a satellite affected by ASI can be optimized to minimize ASI effects.","(This can result in significant savings in satellite power, since without such quantitative tools, unnecessarily large link margins may be used to protect against ASI.)","As described, adding a capability to a CSM system to characterize ASI provides a significant benefit to a satellite system operator.","Such addition would require only modest resources in addition to those required by the basic CSM system.","The features of such a system are readily implemented on the basis of even the high level description that has been provided.","Specifically, they include the incorporation into a CSM of a power spectrum measurement and analysis capability that results in a detailed description of frequency plans used by adjacent satellites.","They also consist of the use of the measured adjacent satellite frequency plans, including: i) Using adjacent satellite frequency plan information, as determined by an enhanced CSM, to identify specific interferers that have been measured in the victim satellite's transponders by the CSM as being due to ASI, and if so, whether it is uplink or downlink ASI.","ii) Using adjacent satellite frequency plan information, as determined by an enhanced CSM, to facilitate inter-system coordination.","iii) Using adjacent satellite frequency plan information, as determined by an enhanced CSM, to verify intra-system ASI coordination provisions.","iv) Using adjacent satellite frequency plan information, as determined by an enhanced CSM, to analyze the effect of ASI on carrier-by-carrier basis for a FDMA transponder.","v) Using adjacent satellite frequency plan information, as determined by an enhanced CSM, to optimize the uplink powers of individual carriers for a EDMA transponder where ASI is significant, so as to maximize the transponder's power-limited capacity.","It should be noted that this improvement over the conventional CSM system will minimize the amount of equipment and software required to measure and characterize ASI, as compared to the standalone systems used conventionally, notwithstanding the enhanced automation that is used.","While the present invention has been explained in accordance with certain preferred or exemplary embodiments, it is not limited thereto, and the scope of the invention is to be defined by the appended claims, as interpreted in accordance with applicable law."]],"string":"[\n [\n \"Method and apparatus for measuring adjacent satellite interference\",\n \"A system for determining at least one of uplink ASI and downlink ASI in a satellite communication system having at least a first satellite and a second satellite, each having uplink and downlink footprints and being in communication with respective earth stations and having a potential interference.\",\n \"The system comprises a first antenna for receiving downlink signals communicated from said first satellite, a second antenna for receiving downlink signals communicated from said second satellite, power spectrum measurement equipment connected to the second antenna for measuring the second satellite power spectrum, and CSM equipment connected to the first antenna and the power spectrum measurement equipment for generating an adjacent satellite carrier plan.\"\n ],\n [\n \"1.A method of characterizing downlink ASI in a communications satellite system having at least a first satellite and at least a second adjacent satellite, each said satellite being in communication with a respective earth station via downlink beams that provide overlapping downlink beam footprints, said method comprising: a) capturing the power spectrum of the second adjacent satellite; b) dividing the downlink beam of the second adjacent satellite into transponder segments; c) performing spectrum analysis on each said transponder segment; d) analyzing the measured power spectrum of the second adjacent satellite to convert measured transponder spectra to estimates of carrier frequency plans.\",\n \"2.The method of characterizing downlink ASI as recited in claim 1, further comprising, determining downlink ASI on a system-wide basis by repeating the process for each downlink beam of the first satellite, and for each of plural adjacent satellites.\",\n \"3.The method characterizing downlink ASI as recited in of claim 1 further comprising using different measurement locations for each of different downlink beams that cover different geographical regions.\",\n \"4.The method of characterizing downlink ASI as recited in claim 1 further comprising pointing a CSM antenna towards the adjacent satellite.\",\n \"5.The method of characterizing downlink ASI as recited in claim 1 further comprising using spectral parameters of power, center frequency and spectral shape, and optionally at least one of modulation type, FEC coding and scrambling patterns.\",\n \"6.A method of characterizing uplink ASI in a communications satellite system having at least a first satellite and a second adjacent satellite, each said satellite being in communication with respective earth stations via downlink beams that provide downlink beam footprints and uplink beams that have uplink beam footprints, said method comprising: a) using the downlink spectrum of the interfering satellite; and b) processing the downlink in accordance with the method of 1) capturing the power spectrum of the second adjacent satellite via spectrum measurement equipment; 2) dividing the downlink beam of the second adjacent satellite into segments; 3) performing spectrum analysis on each said segment; and 4) analyzing the measured power spectrum of the second adjacent satellite to convert measured transponder spectra to estimates of carrier frequency plans.\",\n \"7.The method of characterizing uplink ASI as claimed in claim 6, further comprising: if the second adjacent satellite uses the same uplink and downlink frequency bands as the first satellite, then using only the power spectrum measurement of the second adjacent satellite's downlink spectrum to represent an uplink ASI.\",\n \"8.The method of characterizing uplink ASI as claimed in claim 6, further comprising: if either the uplink or the downlink of the second adjacent satellite operates in frequency bands that are different from the first satellite, determining whether the downlink frequency band is the same, and if the uplink of the second adjacent satellite shares the same frequency band with the first satellite, but where the downlink of the second adjacent satellite uses a different band, receiving the downlink of the second adjacent satellite and characterizing uplink ASI.\",\n \"9.The method of characterizing uplink ASI as claimed in claim 6, further comprising repeating the characterization of uplink ASI on a system-wide basis, by repeating the process for each uplink beam of the first satellite, and for each adjacent satellite.\",\n \"10.A method of characterizing uplink ASI as recited in claim 6 further comprising using the derived information in identifying the source of interference.\",\n \"11.A system for determining at least one of uplink ASI and downlink ASI in a satellite communication system having at least a first satellite and a second satellite, each having uplink and downlink footprints and being in communication with respective earth stations and having a potential interference, comprising: a first antenna for receiving downlink signals communicated from said first satellite; a second antenna for receiving downlink signals communicated from said second satellite; power spectrum measurement equipment connected to said second antenna for measuring the second satellite power spectrum; and CSM equipment connected to said first antenna and said power spectrum measurement equipment for generating an adjacent satellite carrier plan.\",\n \"12.The system as set forth in claim 11 wherein said second antenna is a steerable antenna.\",\n \"13.The system as set forth in claim 11 further comprising at least one of: a) means for providing ASI interference identification; b) means for providing data for inter-system coordination; c) means for providing verification of intra-system ASI provisions; and d) means for system transmission planning.\",\n \"14.The system as set forth in claim 13, wherein said means for system transmission planning comprises: means for analysis of ASI effects on a per carrier basis; and means for carrier by carrier power level optimization.\",\n \"15.The system as set forth in claim 11, further comprising means for determining whether each of the uplink and downlink frequency bands for the first and second satellites are the same and, if the same, using the power spectrum of the downlink to represent uplink ASI.\"\n ],\n [\n \"<SOH> FIELD OF THE INVENTION <EOH>The invention relates generally to a method and apparatus for measuring adjacent satellite interference (ASI) using communications system monitoring equipment, for both up-link and down-link interference.\",\n \"ASI is the interference caused to a victim satellite and one or more of the earth stations with which it is in communication by transmissions from an interfering satellite or the earth stations with which it communicates.\",\n \"Typically, ASI is caused by earth station antenna sidelobes or as a result of antenna mis-pointing.\",\n \"The characterization of adjacent satellite interference (ASI) is a function that is not normally performed by communications system monitoring (CSM) systems implemented in commercial communications satellite networks.\",\n \"However, due to decreased satellite spacing along the geostationary arc, which increases the severity of ASI, it has now become appropriate that the transmissions of adjacent satellites should be monitored, in order to characterize ASI.\",\n \"Such monitoring would permit coordination agreements to be policed, and would allow carrier frequency plans to be optimized to maximize transponder capacity in the presence of ASI.\",\n \"FIG.\",\n \"1A illustrates a typical satellite system, comprising plural geostationary satellites 1 , 2 , 3 4 , which include two satellites 3 , 4 with beams having respective footprints 6 and 7 (spacecraft antenna coverage areas) that overlap on the surface of the earth 8 .\",\n \"An earth station 5 that is located within the two beam footprints 6 , 7 is designed to transmit to satellite 4 , but its signal also is transmitted to adjacent satellite 3 .\",\n \"Uplink ASI occurs when the earth stations like station 5 are transmitting to an adjacent satellite 4 (in the same frequency band) and cause interference to a victim satellite 3 , either via the earth station's antenna sidelobes, or via the main lobe due to antenna mis-pointing.\",\n \"Similarly, earth stations transmitting to satellites 1 and 2 may also cause uplink ASI.\",\n \"FIG.\",\n \"1B also illustrates a typical satellite system, comprising plural geostationary satellites 11 , 12 , 13 14 , which include two satellites 13 , 14 with beams having respective footprints 16 and 17 that overlap on the surface of the earth 8 .\",\n \"An earth station 15 that is located within the two beam footprints 16 , 17 is designed to receive from satellite 13 , but it also receives a signal transmitted by adjacent satellite 14 .\",\n \"Downlink ASI occurs when earth stations like station 15 that receive transmissions from the victim satellite 13 also receive interference from one or more adjacent satellites 14 (and possibly 11 and 12 ), either via the earth station's antenna sidelobes, or as a result of earth station antenna mis-pointing.\",\n \"In uplink ASI, it is the earth station transmissions of the interfering satellite's network that produce the interference, while for downlink ASI, it is the earth station receiving systems of the victim satellite's network that contribute to the interference.\",\n \"In both uplink and downlink ASI, ASI can be produced by uplink and downlink emissions that exceed authorized levels, in addition to the earth station imperfections noted above.\",\n \"In both uplink and downlink ASL it is a combination of closely spaced satellites and the use of small earth station antennas that increases the severity of ASI.\",\n \"Currently, CSM systems include a capability to monitor (modulation type, FEC coding, scrambling patterns, bit error rate (BER), symbol rate estimation, etc.)\",\n \"but do not include the use of equipment for power spectrum measurement and analysis of adjacent satellites.\",\n \"In short, the conventional CSM systems do not provide a capability to monitor ASI, notwithstanding the improved processing capability that new generation CSM systems offer.\",\n \"Accordingly, it is an object of the present invention to provide a method and apparatus for CSM systems, that permit existing systems to be enhanced at a modest cost to include ASI characterization\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The invention involves a method and apparatus for the provision of a CSM enhancement to include ASI characterization.\",\n \"The invention more particularly involves a method of characterizing downlink ASI in a communications satellite system having at least a first satellite and at least a second adjacent satellite, each satellite being in communication with a respective earth station via downlink beams that provide overlapping downlink beam footprints.\",\n \"The downlink ASI determination method comprises: a) capturing the power spectrum of the second adjacent satellite; b) dividing the downlink beam of the second adjacent satellite into transponder segments; c) performing spectrum analysis on each said transponder segment; and d) analyzing the measured power spectrum of the second adjacent satellite to convert measured transponder spectra to estimates of carrier frequency plans.\",\n \"The invention further comprises a method of characterizing uplink ASI in a communications satellite system having at least a first satellite and a second adjacent satellite, each satellite being in communication with respective earth stations via downlink beams that provide downlink beam footprints and uplink beams that have uplink beam footprints.\",\n \"The method comprises a) using the downlink spectrum of the interfering satellite; and b) processing the downlink in accordance with the above method.\",\n \"The invention also is directed to a system for determining at least one of uplink ASI and downlink ASI in a satellite communication system having at least a first satellite and a second satellite, each having uplink and downlink footprints and being in communication with respective earth stations and having a potential interference.\",\n \"The system comprises a first antenna for receiving downlink signals communicated from said first satellite, a second antenna for receiving downlink signals communicated from said second satellite, power spectrum measurement equipment connected to the second antenna for measuring the second satellite power spectrum, and CSM equipment connected to the first antenna and the power spectrum measurement equipment for generating an adjacent satellite carrier plan.\"\n ],\n [\n \"This application claims the benefit of U.S.\",\n \"Provisional Application Ser.\",\n \"No.\",\n \"60/289389, filed on May 8, 2002.FIELD OF THE INVENTION The invention relates generally to a method and apparatus for measuring adjacent satellite interference (ASI) using communications system monitoring equipment, for both up-link and down-link interference.\",\n \"ASI is the interference caused to a victim satellite and one or more of the earth stations with which it is in communication by transmissions from an interfering satellite or the earth stations with which it communicates.\",\n \"Typically, ASI is caused by earth station antenna sidelobes or as a result of antenna mis-pointing.\",\n \"The characterization of adjacent satellite interference (ASI) is a function that is not normally performed by communications system monitoring (CSM) systems implemented in commercial communications satellite networks.\",\n \"However, due to decreased satellite spacing along the geostationary arc, which increases the severity of ASI, it has now become appropriate that the transmissions of adjacent satellites should be monitored, in order to characterize ASI.\",\n \"Such monitoring would permit coordination agreements to be policed, and would allow carrier frequency plans to be optimized to maximize transponder capacity in the presence of ASI.\",\n \"FIG.\",\n \"1A illustrates a typical satellite system, comprising plural geostationary satellites 1, 2, 3 4, which include two satellites 3, 4 with beams having respective footprints 6 and 7 (spacecraft antenna coverage areas) that overlap on the surface of the earth 8.An earth station 5 that is located within the two beam footprints 6, 7 is designed to transmit to satellite 4, but its signal also is transmitted to adjacent satellite 3.Uplink ASI occurs when the earth stations like station 5 are transmitting to an adjacent satellite 4 (in the same frequency band) and cause interference to a victim satellite 3, either via the earth station's antenna sidelobes, or via the main lobe due to antenna mis-pointing.\",\n \"Similarly, earth stations transmitting to satellites 1 and 2 may also cause uplink ASI.\",\n \"FIG.\",\n \"1B also illustrates a typical satellite system, comprising plural geostationary satellites 11, 12, 13 14, which include two satellites 13, 14 with beams having respective footprints 16 and 17 that overlap on the surface of the earth 8.An earth station 15 that is located within the two beam footprints 16, 17 is designed to receive from satellite 13, but it also receives a signal transmitted by adjacent satellite 14.Downlink ASI occurs when earth stations like station 15 that receive transmissions from the victim satellite 13 also receive interference from one or more adjacent satellites 14 (and possibly 11 and 12), either via the earth station's antenna sidelobes, or as a result of earth station antenna mis-pointing.\",\n \"In uplink ASI, it is the earth station transmissions of the interfering satellite's network that produce the interference, while for downlink ASI, it is the earth station receiving systems of the victim satellite's network that contribute to the interference.\",\n \"In both uplink and downlink ASI, ASI can be produced by uplink and downlink emissions that exceed authorized levels, in addition to the earth station imperfections noted above.\",\n \"In both uplink and downlink ASL it is a combination of closely spaced satellites and the use of small earth station antennas that increases the severity of ASI.\",\n \"Currently, CSM systems include a capability to monitor (modulation type, FEC coding, scrambling patterns, bit error rate (BER), symbol rate estimation, etc.)\",\n \"but do not include the use of equipment for power spectrum measurement and analysis of adjacent satellites.\",\n \"In short, the conventional CSM systems do not provide a capability to monitor ASI, notwithstanding the improved processing capability that new generation CSM systems offer.\",\n \"Accordingly, it is an object of the present invention to provide a method and apparatus for CSM systems, that permit existing systems to be enhanced at a modest cost to include ASI characterization SUMMARY OF THE INVENTION The invention involves a method and apparatus for the provision of a CSM enhancement to include ASI characterization.\",\n \"The invention more particularly involves a method of characterizing downlink ASI in a communications satellite system having at least a first satellite and at least a second adjacent satellite, each satellite being in communication with a respective earth station via downlink beams that provide overlapping downlink beam footprints.\",\n \"The downlink ASI determination method comprises: a) capturing the power spectrum of the second adjacent satellite; b) dividing the downlink beam of the second adjacent satellite into transponder segments; c) performing spectrum analysis on each said transponder segment; and d) analyzing the measured power spectrum of the second adjacent satellite to convert measured transponder spectra to estimates of carrier frequency plans.\",\n \"The invention further comprises a method of characterizing uplink ASI in a communications satellite system having at least a first satellite and a second adjacent satellite, each satellite being in communication with respective earth stations via downlink beams that provide downlink beam footprints and uplink beams that have uplink beam footprints.\",\n \"The method comprises a) using the downlink spectrum of the interfering satellite; and b) processing the downlink in accordance with the above method.\",\n \"The invention also is directed to a system for determining at least one of uplink ASI and downlink ASI in a satellite communication system having at least a first satellite and a second satellite, each having uplink and downlink footprints and being in communication with respective earth stations and having a potential interference.\",\n \"The system comprises a first antenna for receiving downlink signals communicated from said first satellite, a second antenna for receiving downlink signals communicated from said second satellite, power spectrum measurement equipment connected to the second antenna for measuring the second satellite power spectrum, and CSM equipment connected to the first antenna and the power spectrum measurement equipment for generating an adjacent satellite carrier plan.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1A is a schematic illustration of a satellite system in which Uplink ASI occurs.\",\n \"FIG.\",\n \"1B is a schematic illustration of a satellite system in which Uplink ASI occurs.\",\n \"FIG.\",\n \"2 is a schematic illustration of a satellite system having a CSM capability that includes ASI detection.\",\n \"FIG.\",\n \"3A is a flow chart that identifies a process for determining downlink ASI.\",\n \"FIG.\",\n \"3B is a flow chart that identifies a process for determining uplink ASI.\",\n \"DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG.\",\n \"2 illustrates the ASI-related features of a satellite system 20 having a plurality of satellites 21, 22, 23, 24 in geostationary orbit, including a satellite that is desired for communication 23 and an adjacent interfering satellite 24.The desired satellite 23 is in communication with an earth station 25 having CSM equipment 28 that can provide conventional CSM functions.\",\n \"Those conventional functions are supplemented by equipment That can determine an adjacent satellite carrier plan.\",\n \"Specifically, a second antenna 26, which may be a shared and steerable antenna that serves several satellites, or a separate dedicated antenna, is provided with equipment that can measure the adjacent satellite 24 power spectrum.\",\n \"That power spectrum output is provided to the CSM 28, which also has the capability 29 to identify the adjacent satellite carrier plan.\",\n \"It also has the capability to characterize ASI for several purposes, including identification of ASI interference 30, provide data for inter-system coordination 31, verify intra-system ASI provisions 32 and utilize system transmission planning software 33 for a variety of purposes.\",\n \"Most importantly, the software can be used to provide an analysis of ASI effects on a per carrier basis 34 and provide carrier by carrier power level optimization 35 in order to ensure maximum capacity in the presence of ASI.\",\n \"The CSM equipment 28 also must have an adequate processing capability, provided by a conventional processor (not shown) with appropriate software modules.\",\n \"Finally, there must be a means for conveying adjacent satellite spectral measurements to the location where ASI processing is performed.\",\n \"Depending on the network architecture employed by the basic CSM system, and the extent to which the required processing functions are centralized, additional provisions may be incorporated to convey ASI power spectrum measurements to the locations where ASI information is further processed and used.\",\n \"While the design of the processing and distribution architecture involves tradeoffs between the extent of local processing that is performed, and the amount of data required to be transported, these considerations would not be major issues for a CSM enhancement, particularly if ASI power spectrum measurements are made infrequently.\",\n \"The antenna 26 that is used to measure emissions from adjacent satellites not only should be able to be pointed along the geostationary arc, but should also be capable of receiving both linear and circular polarizations of each sense, and over bands that may extend beyond the victim satellite's downlink band.\",\n \"If downlink ASI were only of interest, then only the polarization sense(s) and bandwidth of the victim satellite's downlinks need be measured.\",\n \"However if uplink ASI characterization is required, other polarizations and other receive bands may be required to obtain the required replicas of the interfering satellite's uplink spectrum.\",\n \"This requirement is due to the fact that the interfering satellite may use the same uplink bands as the victim satellite, but different downlink bands.\",\n \"Turning next to the application of the system of FIG.\",\n \"2 to the solution of practical problems, is should be noted first that there are differences in uplink and downlink ASI, as further detailed subsequently.\",\n \"However, since the implementation to characterize downlink ASI is somewhat simpler than for uplink ASI, downlink ASI is described first.\",\n \"In downlink ASI, the footprints of both the victim and the interfering satellite overlap in the geographical areas that experience ASI.\",\n \"Also, the downlink spectrum used by affected transponders of the victim satellite overlaps that of the interfering satellite.\",\n \"The objective of downlink ASI characterization is to obtain a description of the interfering satellite's downlink carrier plan in the transponders of the interfering satellite that overlap both the coverage area and frequency band of the victim satellite.\",\n \"Downlink ASI characterization can be used to assist in identifying the source of interference to the victim satellite 23, in the inter-system coordination process with the operator of the adjacent satellite 24, and to verify that the adjacent satellite's frequency plan matches detailed ASI database information (for cases where the adjacent satellite 24 has the same operator as the victim satellite 23).\",\n \"As indicated in function 34 of FIG.\",\n \"2, an important use of the adjacent satellite's carrier frequency plan is to incorporate it into the transmission analysis and planning functions of computing the effects of downlink ASI on a carrier-by-carrier basis.\",\n \"A further function, as indicated in function 35 of FIG.\",\n \"2 is optimizing the carrier frequency plan of the victim satellite to ensure maximum throughput in the presence of ASI.\",\n \"This latter application is useful in cases where either by agreement, or by violation of coordination agreements, ASI is significant enough to require that transmission planning be performed on a carrier-by-carrier basis, rather than for entire bandwidth segments.\",\n \"The procedure to characterize downlink ASI is illustrated in the flowchart of FIG.\",\n \"3A.\",\n \"A first step S1 is to point the CSM antenna 26 (or equivalently, connect to another dedicated antenna whose output can be processed by the CSM 28) towards the adjacent satellite.\",\n \"A second step S2 is to “capture” the power spectrum of the adjacent satellite 24 via the spectrum measurement equipment 27.A third and fourth step S3 and S4 involve the performance of analysis functions in the CSM.\",\n \"For this purpose, spectrum analysis should be done in segments that match the transponder frequency ranges of the victim satellite 23.Accordingly, in step S3, the downlink beam is divided into segments and in step S4 the spectrum analysis is performed on the segment.\",\n \"This measurement must be repeated for all segments until all segments are analyzed.\",\n \"Thus, in step S5, a determination is made as to whether the last segment has been analyzed, and if not, the process loops back to step S4.If it is the last segment, then every transponder segment transmitted by the downlink beam of the interfering satellite 24, which was measured in steps S1 and S2, that causes ASI to one or more downlink beams of the victim satellite 23 has been tested.\",\n \"To characterize downlink ASI on a system wide basis, the process illustrated by FIG.\",\n \"3A is repeated for each downlink beam of the desired satellite 23, and for each of the adjacent satellites (21, 22, and 24).\",\n \"Note that to characterize downlink ASI for different downlink beams of the desired satellite 23, different measurement locations are required for each of the downlink beams since they cover different geographical regions (except for the case of polarization reuse beams, where in some cases, one measurement site may be able to be used for two beams).\",\n \"The measured power spectrum of the adjacent satellite 24 is analyzed in step S6 with software that converts measured transponder spectra to estimates of carrier frequency plans 29.Then, the program ends at step S7.Although for the purposes of ASI interference computation, only the spectral parameters of power, center frequency and spectral shape are required, other parameters such as modulation type, the use of FEC coding, scrambling patterns, etc., that may be outputs of the basic CSM spectral analysis software, also may be used in documenting and coordinating ASI.\",\n \"Although the way in which information on uplink ASI is used is similar to the way information on downlink ASI is used, the uplink ASI generation mechanism is different from downlink ASI, as earlier noted.\",\n \"In uplink ASI, it is earth stations transmitting to an adjacent satellite 24 that are the source of interference.\",\n \"Assuming that the adjacent satellites 24 are of the conventional bent-pipe type (i.e., that the adjacent satellites do not incorporate on-board processing) the way to characterize uplink ASI is to monitor the downlink of the transponder, which has the carriers that generate uplink ASI, in the adjacent satellite 24.Since it is often the case in commercial satellites that the transponder uplink beam coverages do not overlap the downlink beam coverages, the earth stations that measure the spectrum of the adjacent satellite's transponder, whose carriers are the source of uplink ASI, often must be located far from the uplink coverage area where the uplink ASI is produced.\",\n \"Thus, while to characterize downlink ASI, little information is required about the adjacent satellite except for its location and downlink footprint (i.e., all other information relevant to downlink ASI can be measured) more information about the interfering satellite is required to characterize uplink ASI.\",\n \"Specifically, both the uplink and downlink beam footprints, the uplink and downlink frequency bands, and the connectivity between the uplink and the transponders of the interfering satellite must be known, so that the locations of the monitoring earth stations can be determined.\",\n \"Thus, in a flowchart in FIG.\",\n \"3B representing the uplink ASI characterization process, the first step S11 is to determine station location information In step S12, it is then determined if the adjacent satellite 24 uses the same uplink and downlink frequency bands as the victim satellite 23.If so, then every power spectrum measurement of an adjacent satellite's downlink spectrum (on a transponder-by-transponder basis) will represent both an uplink ASI and a downlink ASI spectrum, and such measurement would be used in step S13.However, this is not the case if either the uplink or the downlink of the adjacent satellite 24 operates in frequency bands that are different from the victim satellite 23.A determination is made in step S14 of whether the downlink frequency band is the same.\",\n \"For those cases where the uplink of the interfering satellite 24 shares the same frequency band, but where the downlink of the interfering satellite 24 uses a different band (such as, for example, in the case of a cross-strapped transponder) the downlink of the interfering satellite 24 must be received in order to characterize uplink ASI, even though the downlink is in a band that is different from the downlink bands used by the victim satellite 23.To characterize uplink ASI on a system-wide basis, the process illustrated by FIG.\",\n \"3B is repeated for each uplink beam of the desired satellite, and for each adjacent satellite.\",\n \"Once the uplink ASI power spectrum is measured, its characterization and its use are essentially the same as previously described for downlink ASI.\",\n \"That is, information on uplink ASI can be used to assist in identifying the source of interference, as in function 30.It also can assist is coordinating ASI problems with the operator of the adjacent satellite, as in function 31.A further function 32 is to verify that the adjacent satellite's carrier plan conforms to ASI database information in the case where the adjacent satellite is under control of the same operator.\",\n \"Also, uplink ASI can be used with software 33 to compute the effects of ASI on a carrier-by-carrier basis, as in function 34 and optimize capacity in transponders subject to significant levels of ASI, as in function 35.A transmission planning and analysis program 33 would be implemented in a manner known in the art.\",\n \"The software required for ASI characterization is in two basic categories.\",\n \"First, there is the software required to convert a measured power spectrum into an estimated ASI carrier frequency plan.\",\n \"Second, there is the software that uses the ASI characterization results.\",\n \"It is only the former category that requires processing by the basic CSM equipment, and this processing requirement should be modest since the computations need not be made in real time, and, as noted above, ASI power spectrum measurements should only be required relatively infrequently.\",\n \"The software required for the purposes of using the ASI characterization results is part of the system operations, planning, and inter-system coordination functions.\",\n \"Hence, any hardware associated with these software functions would not be part of the basic CSM system.\",\n \"Returning to the first category of software required for ASI characterization, namely that required to convert the measured adjacent satellite's power spectrum into an estimate of the adjacent satellite's carrier frequency plan (over the portion of bandwidth of interest), there are two subcategories.\",\n \"A first subcategory is software that performs this function for the transponders of the desired satellite 23, as part of a basic CSM system.\",\n \"The requirements for this software are not discussed herein as they are known.\",\n \"However, the second subcategory involves software that defines what ASI measurements are required to be made.\",\n \"This software would use information on adjacent satellites 24 to define the direction, frequency bands, and polarization that are required for ASI measurements.\",\n \"The complexity of the software required for this subcategory is minimal, and would be easily acquired by one skilled in the art, once the requirement is known.\",\n \"If only downlink ASI were being characterized, an alternative “search mode” procedure could be used to search for satellites along the geostationary arc having emissions in the same band and polarization as the downlink of the desired satellite.\",\n \"Assuming that the footprints of the adjacent satellites overlap with the desired satellite, a procedure for downlink characterization would not require the subcategory of measurement control software identified above.\",\n \"Turning now to the second category of software, it includes software that makes use of the estimated adjacent satellite carrier plans.\",\n \"Applications in this category include: a) Software that assists users in identifying interference measured by the CSM for the desired satellite as having ASI, and if so, whether it is uplink or downlink ASI.\",\n \"b) Software that generates adjacent satellite carrier plans in a format useful for intersystem coordination.\",\n \"c) Software that compares a measured adjacent satellite carrier plan against ASI information stored in a database.\",\n \"(Useful for system operators that have multiple adjacent satellites.)\",\n \"d) Transmission planning and analysis software that computes the effects of ASI on the performance of carriers, and optimizes carrier plans to maximize capacity in the presence of ASI.\",\n \"An example of the last category of software would be the well known STRIP7 and COMPLAN programs developed by LMGT (formerly COMSAT Laboratories).\",\n \"The STRIP7 program is specialized to Intelsat's requirements, and has a provision for computing impairments caused by intra-system ASI from its own satellites.\",\n \"The COMPLAN program, which is applicable to non-Intelsat satellites, requires a straightforward modification to handle ASI impairments.\",\n \"These two programs (or software with similar capabilities) perform two important functions for an FDMA (frequency division multiple access) system.\",\n \"The first function is that they analyze the performance of every carrier in a satellite transponder, taking into account impairments due to thermal noise, intermodulation noise, intra-satellite co-channel interference, adjacent carrier interference, and uplink and downlink rain impairments.\",\n \"To these impairments, ASI must be added in the software envisioned for this application (which is already implemented in STRIP7, as noted above).\",\n \"Since the amount of ASI is dependent on both the characteristics of the uplink earth stations of the interfering satellite, and the downlink earth stations of the victim satellite, the ability to analyze transmission performance on an individual carrier basis is important, since such performance estimates cannot be accurately made solely from the carrier plan of the adjacent satellite.\",\n \"The second function of these two transmission analysis programs is that given the carrier frequency assignments of a frequency plan, the uplink carrier powers are optimized so that the power-limited capacity of the transponder is maximized.\",\n \"This is done by computing the effects of the impairments noted above on each individual carrier and providing each carrier with the amount of power it requires to achieve specified performance, while maintaining an optimum transponder operating point (i.e., while maintaining an optimum balance between downlink thermal noise, intermodulation noise, and interference.)\",\n \"The utility of applying ASI to transmission planning and analysis software is that for situations where ASI is significant, and cannot be eliminated by either intra- or inter-system coordination, the effects of ASI can be quantified on an individual carrier basis, and the transmission plan of a satellite affected by ASI can be optimized to minimize ASI effects.\",\n \"(This can result in significant savings in satellite power, since without such quantitative tools, unnecessarily large link margins may be used to protect against ASI.)\",\n \"As described, adding a capability to a CSM system to characterize ASI provides a significant benefit to a satellite system operator.\",\n \"Such addition would require only modest resources in addition to those required by the basic CSM system.\",\n \"The features of such a system are readily implemented on the basis of even the high level description that has been provided.\",\n \"Specifically, they include the incorporation into a CSM of a power spectrum measurement and analysis capability that results in a detailed description of frequency plans used by adjacent satellites.\",\n \"They also consist of the use of the measured adjacent satellite frequency plans, including: i) Using adjacent satellite frequency plan information, as determined by an enhanced CSM, to identify specific interferers that have been measured in the victim satellite's transponders by the CSM as being due to ASI, and if so, whether it is uplink or downlink ASI.\",\n \"ii) Using adjacent satellite frequency plan information, as determined by an enhanced CSM, to facilitate inter-system coordination.\",\n \"iii) Using adjacent satellite frequency plan information, as determined by an enhanced CSM, to verify intra-system ASI coordination provisions.\",\n \"iv) Using adjacent satellite frequency plan information, as determined by an enhanced CSM, to analyze the effect of ASI on carrier-by-carrier basis for a FDMA transponder.\",\n \"v) Using adjacent satellite frequency plan information, as determined by an enhanced CSM, to optimize the uplink powers of individual carriers for a EDMA transponder where ASI is significant, so as to maximize the transponder's power-limited capacity.\",\n \"It should be noted that this improvement over the conventional CSM system will minimize the amount of equipment and software required to measure and characterize ASI, as compared to the standalone systems used conventionally, notwithstanding the enhanced automation that is used.\",\n \"While the present invention has been explained in accordance with certain preferred or exemplary embodiments, it is not limited thereto, and the scope of the invention is to be defined by the appended claims, as interpreted in accordance with applicable law.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476870"}}},{"rowIdx":980,"cells":{"text":{"kind":"list like","value":[["Hinge assembly","A hinge assembly for a foldable working platform, said hinge assembly comprising a hinge body (22), first and second hinge arms (24,26) connected to the hinge body (22), the first hinge arm being rotatable relative to the hinge body about a first axis of rotation (28), the second hinge arm being rotatable relative to the hinge body about a second axis of rotation (30), wherein the first and second axes (28,30) are non-coaxial and wherein movement of the hinge arms beyond the open position is restricted by interaction of abutment portions of the first and second hinge arms with respective abutment portions of the hinge body."],["1.A hinge assembly for a foldable working platform, said platform comprising first and second leaves, such that in use, when the hinge assembly is in an open position, the first leaf is supported in a substantially horizontal, cantilevered configuration by said second leaf via said hinge assembly so that the first and second leaves form a substantially continuous working platform, and when the hinge is in a closed position, the first leaf is arranged substantially parallel to and adjacent the second leaf, so that the working platform is in a folded configuration, said hinge assembly comprising: a hinge body; first and second hinge arms connected to the hinge body; the first hinge arm being rotatable relative to the hinge body about a first axis of rotation; the second hinge arm being rotatable relative to the hinge body about a second axis of rotation; wherein the first and second axes are non-coaxial; and wherein movement of the hinge arms beyond the open position is restricted by interaction of abutment portions of the first and second hinge arms with respective abutment portions of the hinge body.","2.A hinge assembly as claimed in claim 1, wherein the abutment portions comprise complementary arcuate mating surfaces.","3.A hinge assembly as claimed in claim 1, wherein at least one abutment portion on at least one of the hinge arms or the hinge body comprises a step or ridge portion upstanding from a surface of said hinge arm or hinge body.","4.A hinge assembly as claimed in claim 1, wherein at least one of a hinge arm or the hinge body includes a body portion adapted to slide into a cavity provided in the other of said hinge arm and said hinge body as the hinge body and hinge arm rotate relative to one another in order to avoid or reduce the creation and/or termination of gaps between the hinge body and the hinge arm during relative rotation.","5.A hinge assembly as claimed in claim 4, wherein at least one abutment portion comprises a ridge or step upstanding from said body portion.","6.A hinge assembly as claimed in claim 1, wherein an abutment portion is provided on at least two of an inner side, an outer side and a bottom of at least one of the hinge body or hinge arm.","7.A hinge assembly as claimed in claim 1, wherein the hinge body includes an attachment portion for attachment of a cover for a working platform, and in use, said attachment portion serves to help secure the cover.","8.A hinge assembly as claimed in claim 7, wherein said attachment portion is adapted to pass through an aperture in a cover.","9.A hinge assembly as claimed in claim 1, wherein at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform.","10.A hinge assembly as claimed in claim 7, wherein at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform and said attachment portion comprises a guide portion to guide an edge portion of said cover between an attachment point on a working platform and the attachment portion provided on the hinge body.","11.A hinge assembly as claimed in claim 1, wherein the first and second axes of rotation are substantially horizontal in use and respectively pass through the vertical thicknesses of the first and second hinge arms.","12.A hinge assembly as claimed in claim 1, wherein when the hinge assembly is in the open position, longitudinal axes of the first and second hinge arms are substantially co-axial with a longitudinal axis of the hinge body.","13.A hinge assembly as claimed in claim 1, wherein when the hinge assembly is in the closed-position respective longitudinal axes of the first and second hinge arms are substantially parallel, and the longitudinal axes of the first and second hinge arms are substantially perpendicular to a longitudinal axis of the hinge body.","14.A foldable working platform comprising at least one hinge assembly as claimed in claim 1.15.A foldable working platform comprising a plurality of hinge assemblies as claimed in claim 1.16.An ironing board including a working platform as claimed in claim 14."],["<SOH> BACKGROUND OF THE INVENTION <EOH>Foldable working platforms such as, for example, foldable ironing boards, typically include two leaves hinged together and additional supporting elements in the form of frame, leg or strut elements for supporting each of the two leaves.","There is believed to be a need for a support structure for a foldable working platform which reduces the number of supporting elements, the bulk of the supporting elements, and/or the structural or practical complexity of deploying the leaves and/or supporting elements or which at least provides an alternative to known support structures."],["<SOH> SUMMARY OF THE INVENTION <EOH>According to a first aspect of the present invention, there is provided a hinge assembly for a foldable working platform, said platform comprising first and second leaves, such that in use, when the hinge assembly is in an open position, the first leaf is supported in a substantially horizontal, cantilevered configuration by said second leaf via said hinge assembly so that the first and second leaves form a substantially continuous working platform, and when the hinge is in a closed position, the first leaf is arranged substantially parallel to and adjacent the second leaf, so that the working platform is in a folded configuration, said hinge assembly comprising: a hinge body; first and second hinge arms connected to the hinge body; the first hinge arm being rotatable relative to the hinge body about a first axis of rotation; the second hinge arm being rotatable relative to the hinge body about a second axis of rotation; wherein the first and second axes are non-coaxial; and wherein movement of the hinge arms beyond the open position is restricted by interaction of abutment portions of the first and second hinge arms with respective abutment portions of the hinge body.","As the first and second axes of rotation are non-coaxial, the first and second hinge arms can be rotated about the hinge body with clearance between one another to a closed position in which the hinge arms are substantially parallel with one another and hence angularly separated by 0°, which is often difficult or impossible with a conventional hinge in which both hinge arms rotate about a common axis of rotation.","The hinge arms may be elongate members of significant length or may be arranged to be attached to such members enabling the members to be folded adjacent one another in the closed position and extended away from one another in the open position.","A plurality of the hinge assemblies may be used in unison, for example, a pair of the hinge assemblies may be used to support edges of a table top to facilitate folding of one leaf of the table top atop another leaf of the table top when not in use.","Preferably, the abutment portions comprise complementary arcuate mating surfaces.","Preferably, at least one abutment portion on at least one of the hinge arms or the hinge body comprises a step or ridge portion upstanding from a surface of said hinge arm or hinge body.","Preferably, at least one of a hinge arm or the hinge body includes a body portion adapted to slide into a cavity provided in the other of said hinge arm and said hinge body as the hinge body and hinge arm rotate relative to one another in order to avoid or reduce the creation and/or termination of gaps between the hinge body and the hinge arm during relative rotation.","Preferably, at least one abutment portion comprises a ridge or step upstanding from said body portion.","Preferably, an abutment portion is provided on at least two of an inner side, an outer side and a bottom of at least one of the hinge body or hinge arm.","Preferably, the hinge body includes an attachment portion for attachment of a cover for a working platform, and in use, said attachment portion serves to help secure the cover.","Preferably, said attachment portion is adapted to pass through an aperture in a cover.","Preferably, at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform.","Preferably, at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform and said attachment portion comprises a guide portion to guide an edge portion of said cover between an attachment point on a working platform and the attachment portion provided on the hinge body.","Preferably, the first and second axes of rotation are substantially horizontal in use and respectively pass through the vertical thicknesses of the first and second hinge arms.","Preferably, when the hinge assembly is in the open position, longitudinal axes of the first and second hinge arms are substantially co-axial with a longitudinal axis of the hinge body.","Preferably, when the hinge assembly is in the closed-position respective longitudinal axes of the first and second hinge arms are substantially parallel, and the longitudinal axes of the first and second hinge arms are substantially perpendicular to a longitudinal axis of the hinge body.","Further aspects of the present invention provide a working platform comprising first and second leaves mutually connected by one or more hinge assemblies in accordance with the first aspect.","A further aspect of the present invention provides an ironing board comprising first and second leaves mutually connected by one or more hinge assemblies in accordance with the first aspect.","In a particularly preferred embodiment of the present invention, a pair of hinge assemblies connect two leaves of the working platform of a folding ironing board.","Preferably, the ironing board can be folded approximately centrally of the longitudinal extent of its working platform."],["FIELD OF THE INVENTION The present invention relates to a hinge assembly and especially to a hinge assembly for providing cantilevered support to an otherwise unsupported leaf of a foldable working platform.","BACKGROUND OF THE INVENTION Foldable working platforms such as, for example, foldable ironing boards, typically include two leaves hinged together and additional supporting elements in the form of frame, leg or strut elements for supporting each of the two leaves.","There is believed to be a need for a support structure for a foldable working platform which reduces the number of supporting elements, the bulk of the supporting elements, and/or the structural or practical complexity of deploying the leaves and/or supporting elements or which at least provides an alternative to known support structures.","SUMMARY OF THE INVENTION According to a first aspect of the present invention, there is provided a hinge assembly for a foldable working platform, said platform comprising first and second leaves, such that in use, when the hinge assembly is in an open position, the first leaf is supported in a substantially horizontal, cantilevered configuration by said second leaf via said hinge assembly so that the first and second leaves form a substantially continuous working platform, and when the hinge is in a closed position, the first leaf is arranged substantially parallel to and adjacent the second leaf, so that the working platform is in a folded configuration, said hinge assembly comprising: a hinge body; first and second hinge arms connected to the hinge body; the first hinge arm being rotatable relative to the hinge body about a first axis of rotation; the second hinge arm being rotatable relative to the hinge body about a second axis of rotation; wherein the first and second axes are non-coaxial; and wherein movement of the hinge arms beyond the open position is restricted by interaction of abutment portions of the first and second hinge arms with respective abutment portions of the hinge body.","As the first and second axes of rotation are non-coaxial, the first and second hinge arms can be rotated about the hinge body with clearance between one another to a closed position in which the hinge arms are substantially parallel with one another and hence angularly separated by 0°, which is often difficult or impossible with a conventional hinge in which both hinge arms rotate about a common axis of rotation.","The hinge arms may be elongate members of significant length or may be arranged to be attached to such members enabling the members to be folded adjacent one another in the closed position and extended away from one another in the open position.","A plurality of the hinge assemblies may be used in unison, for example, a pair of the hinge assemblies may be used to support edges of a table top to facilitate folding of one leaf of the table top atop another leaf of the table top when not in use.","Preferably, the abutment portions comprise complementary arcuate mating surfaces.","Preferably, at least one abutment portion on at least one of the hinge arms or the hinge body comprises a step or ridge portion upstanding from a surface of said hinge arm or hinge body.","Preferably, at least one of a hinge arm or the hinge body includes a body portion adapted to slide into a cavity provided in the other of said hinge arm and said hinge body as the hinge body and hinge arm rotate relative to one another in order to avoid or reduce the creation and/or termination of gaps between the hinge body and the hinge arm during relative rotation.","Preferably, at least one abutment portion comprises a ridge or step upstanding from said body portion.","Preferably, an abutment portion is provided on at least two of an inner side, an outer side and a bottom of at least one of the hinge body or hinge arm.","Preferably, the hinge body includes an attachment portion for attachment of a cover for a working platform, and in use, said attachment portion serves to help secure the cover.","Preferably, said attachment portion is adapted to pass through an aperture in a cover.","Preferably, at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform.","Preferably, at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform and said attachment portion comprises a guide portion to guide an edge portion of said cover between an attachment point on a working platform and the attachment portion provided on the hinge body.","Preferably, the first and second axes of rotation are substantially horizontal in use and respectively pass through the vertical thicknesses of the first and second hinge arms.","Preferably, when the hinge assembly is in the open position, longitudinal axes of the first and second hinge arms are substantially co-axial with a longitudinal axis of the hinge body.","Preferably, when the hinge assembly is in the closed-position respective longitudinal axes of the first and second hinge arms are substantially parallel, and the longitudinal axes of the first and second hinge arms are substantially perpendicular to a longitudinal axis of the hinge body.","Further aspects of the present invention provide a working platform comprising first and second leaves mutually connected by one or more hinge assemblies in accordance with the first aspect.","A further aspect of the present invention provides an ironing board comprising first and second leaves mutually connected by one or more hinge assemblies in accordance with the first aspect.","In a particularly preferred embodiment of the present invention, a pair of hinge assemblies connect two leaves of the working platform of a folding ironing board.","Preferably, the ironing board can be folded approximately centrally of the longitudinal extent of its working platform.","BRIEF DESCRIPTION OF THE DRAWINGS Preferred embodiments of the present invention will now be described, by way of example only, with reference to the accompanying drawings, in which: FIG.","1 is a perspective view of a hinge assembly in a fully open position; FIGS.","2 to 4 are perspective views of the hinge assembly of FIG.","1 in sequential intermediate positions in movement between a fully open position and a fully closed position; FIG.","5 is a perspective view of the hinge assembly of FIG.","1 in the fully closed position; FIG.","6 is a partial perspective view illustrating the hinge assembly of FIG.","1 in the fully open position and supporting a leaf of a two-piece working platform of a folding ironing board; FIGS.","7 to 9 are partial perspective views of the hinge assembly and working platform of FIG.","6 in sequential intermediate positions in movement between the fully open position and the fully closed position; FIG.","10 is a partial perspective view of the hinge assembly and working platform of FIG.","6 in the fully closed position; FIGS.","11 to 13 are perspective views of the hinge assembly of FIG.","1 in positions equivalent to those shown in FIGS.","1, 3 and 5 respectively, illustrating the interaction in use of the hinge assembly with a cloth ironing board cover, during movement between the fully open position and the fully closed position, with the working platform omitted for simplicity; FIGS.","14a and 14b are perspective views of a variation of a hinge arm; FIGS.","14c, 14d, 14e and 14f are respectively outer side, bottom, top and end views of the hinge arm of FIGS.","14a and 14b; FIGS.","14g, 14h and 14i are cross-sectional views corresponding to the section lines G-G, H-H and I-I of FIG.","14d; FIGS.","15a and 15b are perspective views of a hinge body for use in the embodiment of FIG.","1; FIGS.","15c, 15d, 15e, 15f and 15g are outer side, bottom, top, inner side, and end views of the hinge body of FIGS.","15a and 15b; and FIG.","15h is a cross-sectional view on H-H of FIG.","15e.","DETAILED DESCRIPTION OF THE DRAWINGS Referring firstly to FIG.","1, the principal elements of a hinge assembly 20 are a hinge body 22, a first hinge arm 24 and a second hinge arm 26.Hinge arms 24 and 26 are pivotally connected to hinge body 22 by first and second hinge pins 28, 30, respectively.","The hinge pins 28 and 30 are spaced apart from one another and substantially parallel.","The respective axes of rotation of the first and second hinge arms 24, 26 relative to the hinge body 22 are thus non-coaxial.","As illustrated in FIG.","1, the hinge assembly 20 is in a fully open position with the hinge arms 24, 26 extending horizontally away from the hinge body 22 in substantially opposing directions and being separated from one another by an angle of 180°.","The first hinge arm 24 may be attached to a first leaf of a working platform, and the second hinge arm 26 may be attached to a second leaf of the working platform.","As illustrated in the drawings, the first leaf could be supported by the second leaf via the hinge assembly (or a plurality of hinge assemblies) and the second leaf would be supported by external support elements such as frame, leg or strut elements.","In movement from the open position illustrated in FIG.","1 to the closed position illustrated in FIG.","5, elements of the hinge assembly 20 rotate relative to each other first about the first hinge pin 28 and subsequently about the second hinge pin 30.Urging the distal end of the first hinge arm 24 upwardly results in its rotation relative to hinge body 22 about the first hinge pin 28 through the position illustrated in FIG.","2.Continued rotation of the first hinge arm 24 results in closed-position mating surfaces 32 of the first hinge arm 24 meeting closed-position mating surfaces 34 of the hinge body 22 at junction 36 at the position illustrated in FIG.","3.At this position, it can be seen that the first hinge arm 24 is generally perpendicular to the hinge body 22.Further rotation about hinge pin 28 is inhibited by the engagement of the closed-position mating surfaces 32, 36 and continued urging of the first hinge arm 24 causes the hinge body 22 to rotate relative to the second hinge arm 26, about the second hinge pin 30, through the position illustrated in FIG.","4.Continued rotation of hinge body 22 about the second hinge pin 30 results in closed-position mating surfaces 38 of the hinge body 22 meeting closed-position mating surfaces 40 of the second hinge arm 26 at junction 42 at the closed position of the hinge assembly illustrated in FIG.","5.In the closed position, the hinge arms 24 and 26 extend horizontally away from the hinge body 22 substantially parallel to one another, and substantially perpendicular to the hinge body 22.This illustrated order of relative movement is preferred, but not essential, and may vary, even in one embodiment due to factors such as wear or lubrication of different parts of the hinge assembly.","Referring now to FIG.","6, the first and second hinge arms 24, 26 are arranged to be received in respective first and second channels 44, 46 of an ironing board working platform 48 and secured thereto by locking tabs 50 which pop into complimentary slots or apertures in the channels 44 and 46.The working platform 48 is formed of first and second leaves 52, 54 which are secured together by a pair of hinge assemblies 20 (only one illustrated).","The leaves 52 and 54 are illustrated in a working (open) position in FIG.","6.By urging the first leaf 52, and by the consequent action of the hinge assemblies 20, the first leaf 52 is rotated relative to the second leaf through the positions illustrated in FIGS.","7 to 9 until the hinge assembly is in its closed position and the first leaf 52 is in a storage position, illustrated in FIG.","10.Only the second leaf 54 of working platform 48 is supported by external support elements 55.In the working position, illustrated in FIG.","6, the first leaf 52 is supported solely by the second leaf via the hinge assemblies 20.In the working (open) position, the first leaf 52 is therefore supported in a cantilevered configuration.","Referring back to FIGS.","1 to 5, the support is provided by interaction between two pairs of open-position mating surfaces.","A first pair of open-position mating surfaces comprises open-position mating surfaces 56 of the first hinge arm 24 and open-position mating surfaces 58 of the hinge body 22.A second pair of open-position mating surfaces comprises open-position mating surfaces 60 of the second hinge arm 26 and open-position mating surfaces 62 of the hinge body 22.The open-position mating surfaces 56, 58, 60, 62 of each pair abut when the hinge assembly is in the open position, illustrated in FIG.","1.The interaction between the mating surfaces 56, 58, 60, 62 is sufficient to support a working load on the first leaf 52 in the working position of FIG.","6.The edges 64 and 66 of leaves 52 and 54 respectively are formed with curved lips 68 and 70 respectively which may be guided by leaf guide 72 of hinge body 22 during movement between the positions illustrated in FIGS.","6 and 10, to maintain the pair of hinge assemblies 20 rotating in unison and thereby minimise the tendency of the edges 64 and 66 to twist relative to one another.","In use, the leaves 52 and 54 will typically be covered by a thin sheet of foam (not shown) and a material cover 74 (see FIGS.","11-14).","Each hinge body 22 is formed with a cover attachment portion in the form of button 76 which retains the material cover 74 by a button hole 78 which maintains the cover 74 taut in the operating position of FIG.","11 and prevents the cover 74 coming away from leaves 52 and 54 in the storage position, illustrated in FIG.","13.Each hinge arm 24 and 26 is formed with a guide tab 80 which guides movement of an elasticised edge 82 of the material cover 74 during rotation of the hinge assemblies 20.The main elements of the hinge assembly will now be described in more detail.","With reference to FIGS.","14a to 14i, the illustrated hinge arm corresponds to, but is slightly different to the second hinge arm 26 of FIGS.","1 to 13 since it is not provided with locking tabs (50 in FIG.","1).","However, since the embodiment of FIGS.","14a to 14i has many common features to that of the embodiment of FIGS.","1 to 13, corresponding reference numerals will be used for corresponding features.","The hinge arm 26 of FIGS.","14a to 14i has an elongate fitting portion 200 for fitting into a channel of an ironing board working platform, and a connection portion 250 for connection and engagement with the hinge body 22 of a hinge assembly 20.The fitting portion 200 has a rearmost end distal from the connection portion 250 and a foremost end connected to the connection portion 250.The fitting portion 200 is generally I-shaped in cross-section, as best shown in FIG.","14h.","The hinge arm 26 may be considered to have an outer side, which in use, faces away from the working platform, and an inner side which, in use, faces towards the centre of the working platform.","On the outer side of the connection portion 250 is located the guide tab 80 which provides a generally downwardly facing guide surface 280 in order to guide movement of an elasticised edge of a material cover (shown in FIGS.","11 to 13).","The guide surfaces are inclined and run in a direction which is generally aligned from the bottom (or non-working surface) extreme of the hinge arm towards the button.","The connection portion 250 includes a cavity 252 for receiving an end portion of a hinge body 22, the cavity 252 being defined by opposing side wall portions 254, 256.Each of the opposing wall portions 254, 256 has a respective circular aperture 255, 257 therein for receiving a hinge pin 30 in order to pivotally attach the hinge arm 26 to the hinge body 22.The opposing wall portions 254, 256 have respective front edges 260, 262 which are arcuate and which have a centre of curvature located at the centre of the respective apertures 255, 257.In use, the front edges 260, 262 run close to or bear against corresponding surfaces of the hinge body 22 (described in detail hereafter) in a configuration which helps avoid catching of objects between the hinge arm 26 and the hinge body 22 during operation, and which may provide support for the hinge pins if required.","The opposing wall portions 254, 256 are connected together by a bottom wall portion 264 and a top wall portion 266.As shown in FIG.","14c, the bottom wall portion 264 extends forwardly from the foremost end of the fitting portion 200 to a position directly beneath the centre of the apertures 255, 257.As is best shown in FIGS.","14a, 14b, 14f and 14g the bottom wall is arched downwardly at its foremost part providing an arcuate inner surface 265.The arcuate foremost edge of the bottom wall 264 provides the open-position mating surfaces 60 of the second hinge arm 26.The top wall 266 is relatively short, extending forwardly from the foremost end of the fitting portion 200 approximately half as far as the bottom wall 264 extends.","The top edges of the wall portions 254, 256 provide the closed-position mating surfaces 38 of the second hinge arm.","(A foremost edge 268 of the top wall 266 may also provide a closed-position mating surface 38 of the second hinge arm, by engagement with the top surface of the hinge body 22, if required.)","The preferred embodiment of a hinge arm is made from aluminium and may conveniently be formed from two separately formed longitudinal “halves”, so that the finished hinge arm has a longitudinal joint line along its length, approximately half way between the inner and outer sides thereof.","Although only one hinge arm 26 is described in detail, and illustrated in FIG.","14, it will be appreciated that the first hinge arm 24 corresponds, but is a mirror image thereof.","With reference to FIGS.","15a to 15h, the preferred embodiment of the hinge body 22 will now be described in detail.","The hinge body 22 includes first and second hinge body end portions 302, 304 at respective first and second ends thereof.","Each end portion 302, 304 is laterally narrow compared to the rest of the hinge body 22 and includes a respective circular aperture 306, 308 therethrough for receiving a respective hinge pin 28, 30 and thereby connecting the hinge body 22 to respective hinge arms 24, 26.Each end portion 302, 304 has a respective arcuate edge portion 310, 312, being approximately quarter circular, extending from an endmost edge part of the respective end portion to a bottommost edge of the respective end portion, and having the respective apertures 306, 308 as their respective centres of curvature.","The arcuate edge portions 310, 312 have rounded surfaces in the lateral direction of the body end portions 302, 304, so that they can slide neatly into and out of the spaces defined by the arcuate inner surfaces 265 of the hinge arms 24, 26, when the hinge body 22 and hinge arms 24, 26 are rotated relative to each other.","This sliding action avoids the creation and termination of gaps, in which objects could get caught, during operation of the hinge assembly.","Intermediate the two hinge body end portions 302, 304 there is a central body portion 314 which has a first side projecting portion 314a projecting to an outer lateral side of the hinge body 22 and a second side projecting portion 314b projecting to an inner lateral side of the hinge body 22.The first and second side portions 314a, 314b of the central body portion 314 meet at the bottom of the hinge body 22 so that the central body portion 314 effectively wraps around the sides and bottom of the hinge body 22.The projecting central body portion 314 provides a number of step or ridge portions which form narrow mating or bearing surfaces for interaction with surfaces on the hinge arms 24, 26.In this embodiment, the mating or bearing surfaces provided project substantially perpendicular to the adjacent surfaces of the end portions 302, 304 of the hinge body 22.Because the projecting central body portion 314 extends substantially symmetrically about both sides of the hinge body 22, similar mating or bearing surfaces are provided on the inner and outer sides of the hinge body 22.For convenience only those provided on the outer side will be described in detail below, and the reference numerals used will be suffixed by the letter “a”.","Corresponding mating surfaces provided on the inner side of the hinge body 22 are designated in the drawings by the same reference numerals but with the suffix “b” replacing the suffix “a”.","Towards the top of the hinge body (as shown, for example in FIG.","15a) the central body portion is relatively small in longitudinal extent and provides substantially straight, substantially parallel, substantially vertical closed-position mating surfaces 34a, 38a, which are adapted to abut mating surfaces 32, 40 of the hinge arms formed by the top edges of the side wall portions 254, 256.It is noted for clarity that directional descriptors to the top, bottom and the like of certain elements are used for convenience despite the rotation of those elements in a vertical plane, in use.","Such directional descriptors herein refer to the relative directions when the elements are in the orientation shown in FIGS.","1, 6 and 11, that is with the hinge assembly 20 fully open and the working platform in a substantially horizontal configuration, and should be construed accordingly.","The projecting central body portion 314 increases in longitudinal extent as it extends downwards (from the bottom of the closed-position mating surfaces 34a, 38a) to provide arcuate bearing surfaces 316a, 318a which have centres of curvature corresponding to the centres of the apertures 306, 308.The arcuate bearing surfaces 316a, 318a extend from the bottom of the vertical mating surfaces 34a, 38a through a quarter circle to respective points substantially vertically below the centres of the apertures 306, 308.These arcuate bearing surfaces 316a, 318a (and corresponding 316b, 318b on the inner side of the hinge body 22) are adapted in use to slide against, or close to, the arcuate edges 260, 262 of the hinge arms, as discussed above.","Also formed by the central body portion 314 where it passes around the underside of the hinge body 22 and extends between the inner and outer sides of the hinge body 22 are open-position mating surfaces 58, 62.These take the form, as best illustrated in FIGS.","15a, 15b and 15g, of vertically orientated semi-annular surfaces.","In use, these open-position mating surfaces 58, 62 are adapted to abut the open-position mating surfaces 56, 60 provided on the hinge arms 24, 26.On the outer side of the central body portion 314a is provided the button 76.On the inner side of the central body portion 314b is provided the leaf guide 72 which provides first and second arcuate guide surfaces 320, 322 adapted to bear against edges of the two leaves.","The leaf guide 72 is provided in order to guide respective leaves of a working platform and help prevent relative twisting of the two leaves which, in practice, would normally be connected by a pair of hinge assemblies 20.The hinge body 22 is preferably die cast from aluminium.","It will be appreciated that the hinge assembly 20 described has a number of beneficial features.","It allows a working platform to be provided in which only one of the leaves needs to be supported by external support elements, since the externally supported leaf can support the other leaf in cantilevered configuration via the hinge assembly or assemblies 20.This provides convenient compact support, and great ease of use as no deployment of external support elements is required.","It is normally preferred to use a pair of hinge assemblies to connect two leaves but in certain applications, for example where the working platform is relatively narrow and provided with a central channel, use of a single hinge assembly might be appropriate.","In the fully open position, the described hinge assembly fits entirely within the thickness (or height) of the hinge arms 24, 26 and this is advantageous over the use of hinge assemblies which protrude above or below the hinge arms and the platform.","When the hinge assembly is in its closed position, the only part of the hinge assembly exposed at the folded terminus of the working platform is the bottom surface of the hinge body 22, providing a blunt and safe platform end, unlike some other hinge assemblies.","Rotation of the hinge arms about two non-coaxial axes of rotation allows first and second leaves to be stored parallel and adjacent each other.","Different thicknesses of working platform or working platform cover (for example, foam padding and/or an ironing board cover) can be accommodated by use of different lengths of hinge body 22.The use of snug-fitting arcuate sliding portions on the hinge body 22 and hinge arms 24, 26 provides additional support to the structure (if this is required due to wear and or play in the hinge pins 28, 30) and minimises the likelihood of items being caught in gaps which vary in size as the hinge assembly is operated.","The provision of closed-position mating surfaces 32, 34, 38, 40 to keep the hinge arms (or leaves) substantially parallel when the, hinge assemblies are closed reduces the likelihood of items, such as fingers, being damaged by being caught between the leaves as the platform is folded.","Although desirable these may not always be necessary, as under some circumstances it may be adequate to allow the leaves of the working surface to contact each other in the closed position.","The provision of a button 76 or other suitable cover retaining means on the hinge body 22 helps to retain the cover in position during folding and unfolding.","Provision of guide tabs 80 further help to retain the cover and prevent it from becoming caught in the hinge assembly.","Throughout this specification and the claims, the words “comprise”, “comprises” and “comprising” are used in a non-exclusive sense, except where the context requires otherwise.","It is to be understood that, if any prior art publication is referred to herein, such reference does not constitute an admission that the publication forms a part of the common general knowledge in the art, in Australia or in any other country.","Modifications and improvements may be incorporated without departing from the scope of the present invention."]],"string":"[\n [\n \"Hinge assembly\",\n \"A hinge assembly for a foldable working platform, said hinge assembly comprising a hinge body (22), first and second hinge arms (24,26) connected to the hinge body (22), the first hinge arm being rotatable relative to the hinge body about a first axis of rotation (28), the second hinge arm being rotatable relative to the hinge body about a second axis of rotation (30), wherein the first and second axes (28,30) are non-coaxial and wherein movement of the hinge arms beyond the open position is restricted by interaction of abutment portions of the first and second hinge arms with respective abutment portions of the hinge body.\"\n ],\n [\n \"1.A hinge assembly for a foldable working platform, said platform comprising first and second leaves, such that in use, when the hinge assembly is in an open position, the first leaf is supported in a substantially horizontal, cantilevered configuration by said second leaf via said hinge assembly so that the first and second leaves form a substantially continuous working platform, and when the hinge is in a closed position, the first leaf is arranged substantially parallel to and adjacent the second leaf, so that the working platform is in a folded configuration, said hinge assembly comprising: a hinge body; first and second hinge arms connected to the hinge body; the first hinge arm being rotatable relative to the hinge body about a first axis of rotation; the second hinge arm being rotatable relative to the hinge body about a second axis of rotation; wherein the first and second axes are non-coaxial; and wherein movement of the hinge arms beyond the open position is restricted by interaction of abutment portions of the first and second hinge arms with respective abutment portions of the hinge body.\",\n \"2.A hinge assembly as claimed in claim 1, wherein the abutment portions comprise complementary arcuate mating surfaces.\",\n \"3.A hinge assembly as claimed in claim 1, wherein at least one abutment portion on at least one of the hinge arms or the hinge body comprises a step or ridge portion upstanding from a surface of said hinge arm or hinge body.\",\n \"4.A hinge assembly as claimed in claim 1, wherein at least one of a hinge arm or the hinge body includes a body portion adapted to slide into a cavity provided in the other of said hinge arm and said hinge body as the hinge body and hinge arm rotate relative to one another in order to avoid or reduce the creation and/or termination of gaps between the hinge body and the hinge arm during relative rotation.\",\n \"5.A hinge assembly as claimed in claim 4, wherein at least one abutment portion comprises a ridge or step upstanding from said body portion.\",\n \"6.A hinge assembly as claimed in claim 1, wherein an abutment portion is provided on at least two of an inner side, an outer side and a bottom of at least one of the hinge body or hinge arm.\",\n \"7.A hinge assembly as claimed in claim 1, wherein the hinge body includes an attachment portion for attachment of a cover for a working platform, and in use, said attachment portion serves to help secure the cover.\",\n \"8.A hinge assembly as claimed in claim 7, wherein said attachment portion is adapted to pass through an aperture in a cover.\",\n \"9.A hinge assembly as claimed in claim 1, wherein at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform.\",\n \"10.A hinge assembly as claimed in claim 7, wherein at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform and said attachment portion comprises a guide portion to guide an edge portion of said cover between an attachment point on a working platform and the attachment portion provided on the hinge body.\",\n \"11.A hinge assembly as claimed in claim 1, wherein the first and second axes of rotation are substantially horizontal in use and respectively pass through the vertical thicknesses of the first and second hinge arms.\",\n \"12.A hinge assembly as claimed in claim 1, wherein when the hinge assembly is in the open position, longitudinal axes of the first and second hinge arms are substantially co-axial with a longitudinal axis of the hinge body.\",\n \"13.A hinge assembly as claimed in claim 1, wherein when the hinge assembly is in the closed-position respective longitudinal axes of the first and second hinge arms are substantially parallel, and the longitudinal axes of the first and second hinge arms are substantially perpendicular to a longitudinal axis of the hinge body.\",\n \"14.A foldable working platform comprising at least one hinge assembly as claimed in claim 1.15.A foldable working platform comprising a plurality of hinge assemblies as claimed in claim 1.16.An ironing board including a working platform as claimed in claim 14.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Foldable working platforms such as, for example, foldable ironing boards, typically include two leaves hinged together and additional supporting elements in the form of frame, leg or strut elements for supporting each of the two leaves.\",\n \"There is believed to be a need for a support structure for a foldable working platform which reduces the number of supporting elements, the bulk of the supporting elements, and/or the structural or practical complexity of deploying the leaves and/or supporting elements or which at least provides an alternative to known support structures.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>According to a first aspect of the present invention, there is provided a hinge assembly for a foldable working platform, said platform comprising first and second leaves, such that in use, when the hinge assembly is in an open position, the first leaf is supported in a substantially horizontal, cantilevered configuration by said second leaf via said hinge assembly so that the first and second leaves form a substantially continuous working platform, and when the hinge is in a closed position, the first leaf is arranged substantially parallel to and adjacent the second leaf, so that the working platform is in a folded configuration, said hinge assembly comprising: a hinge body; first and second hinge arms connected to the hinge body; the first hinge arm being rotatable relative to the hinge body about a first axis of rotation; the second hinge arm being rotatable relative to the hinge body about a second axis of rotation; wherein the first and second axes are non-coaxial; and wherein movement of the hinge arms beyond the open position is restricted by interaction of abutment portions of the first and second hinge arms with respective abutment portions of the hinge body.\",\n \"As the first and second axes of rotation are non-coaxial, the first and second hinge arms can be rotated about the hinge body with clearance between one another to a closed position in which the hinge arms are substantially parallel with one another and hence angularly separated by 0°, which is often difficult or impossible with a conventional hinge in which both hinge arms rotate about a common axis of rotation.\",\n \"The hinge arms may be elongate members of significant length or may be arranged to be attached to such members enabling the members to be folded adjacent one another in the closed position and extended away from one another in the open position.\",\n \"A plurality of the hinge assemblies may be used in unison, for example, a pair of the hinge assemblies may be used to support edges of a table top to facilitate folding of one leaf of the table top atop another leaf of the table top when not in use.\",\n \"Preferably, the abutment portions comprise complementary arcuate mating surfaces.\",\n \"Preferably, at least one abutment portion on at least one of the hinge arms or the hinge body comprises a step or ridge portion upstanding from a surface of said hinge arm or hinge body.\",\n \"Preferably, at least one of a hinge arm or the hinge body includes a body portion adapted to slide into a cavity provided in the other of said hinge arm and said hinge body as the hinge body and hinge arm rotate relative to one another in order to avoid or reduce the creation and/or termination of gaps between the hinge body and the hinge arm during relative rotation.\",\n \"Preferably, at least one abutment portion comprises a ridge or step upstanding from said body portion.\",\n \"Preferably, an abutment portion is provided on at least two of an inner side, an outer side and a bottom of at least one of the hinge body or hinge arm.\",\n \"Preferably, the hinge body includes an attachment portion for attachment of a cover for a working platform, and in use, said attachment portion serves to help secure the cover.\",\n \"Preferably, said attachment portion is adapted to pass through an aperture in a cover.\",\n \"Preferably, at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform.\",\n \"Preferably, at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform and said attachment portion comprises a guide portion to guide an edge portion of said cover between an attachment point on a working platform and the attachment portion provided on the hinge body.\",\n \"Preferably, the first and second axes of rotation are substantially horizontal in use and respectively pass through the vertical thicknesses of the first and second hinge arms.\",\n \"Preferably, when the hinge assembly is in the open position, longitudinal axes of the first and second hinge arms are substantially co-axial with a longitudinal axis of the hinge body.\",\n \"Preferably, when the hinge assembly is in the closed-position respective longitudinal axes of the first and second hinge arms are substantially parallel, and the longitudinal axes of the first and second hinge arms are substantially perpendicular to a longitudinal axis of the hinge body.\",\n \"Further aspects of the present invention provide a working platform comprising first and second leaves mutually connected by one or more hinge assemblies in accordance with the first aspect.\",\n \"A further aspect of the present invention provides an ironing board comprising first and second leaves mutually connected by one or more hinge assemblies in accordance with the first aspect.\",\n \"In a particularly preferred embodiment of the present invention, a pair of hinge assemblies connect two leaves of the working platform of a folding ironing board.\",\n \"Preferably, the ironing board can be folded approximately centrally of the longitudinal extent of its working platform.\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention relates to a hinge assembly and especially to a hinge assembly for providing cantilevered support to an otherwise unsupported leaf of a foldable working platform.\",\n \"BACKGROUND OF THE INVENTION Foldable working platforms such as, for example, foldable ironing boards, typically include two leaves hinged together and additional supporting elements in the form of frame, leg or strut elements for supporting each of the two leaves.\",\n \"There is believed to be a need for a support structure for a foldable working platform which reduces the number of supporting elements, the bulk of the supporting elements, and/or the structural or practical complexity of deploying the leaves and/or supporting elements or which at least provides an alternative to known support structures.\",\n \"SUMMARY OF THE INVENTION According to a first aspect of the present invention, there is provided a hinge assembly for a foldable working platform, said platform comprising first and second leaves, such that in use, when the hinge assembly is in an open position, the first leaf is supported in a substantially horizontal, cantilevered configuration by said second leaf via said hinge assembly so that the first and second leaves form a substantially continuous working platform, and when the hinge is in a closed position, the first leaf is arranged substantially parallel to and adjacent the second leaf, so that the working platform is in a folded configuration, said hinge assembly comprising: a hinge body; first and second hinge arms connected to the hinge body; the first hinge arm being rotatable relative to the hinge body about a first axis of rotation; the second hinge arm being rotatable relative to the hinge body about a second axis of rotation; wherein the first and second axes are non-coaxial; and wherein movement of the hinge arms beyond the open position is restricted by interaction of abutment portions of the first and second hinge arms with respective abutment portions of the hinge body.\",\n \"As the first and second axes of rotation are non-coaxial, the first and second hinge arms can be rotated about the hinge body with clearance between one another to a closed position in which the hinge arms are substantially parallel with one another and hence angularly separated by 0°, which is often difficult or impossible with a conventional hinge in which both hinge arms rotate about a common axis of rotation.\",\n \"The hinge arms may be elongate members of significant length or may be arranged to be attached to such members enabling the members to be folded adjacent one another in the closed position and extended away from one another in the open position.\",\n \"A plurality of the hinge assemblies may be used in unison, for example, a pair of the hinge assemblies may be used to support edges of a table top to facilitate folding of one leaf of the table top atop another leaf of the table top when not in use.\",\n \"Preferably, the abutment portions comprise complementary arcuate mating surfaces.\",\n \"Preferably, at least one abutment portion on at least one of the hinge arms or the hinge body comprises a step or ridge portion upstanding from a surface of said hinge arm or hinge body.\",\n \"Preferably, at least one of a hinge arm or the hinge body includes a body portion adapted to slide into a cavity provided in the other of said hinge arm and said hinge body as the hinge body and hinge arm rotate relative to one another in order to avoid or reduce the creation and/or termination of gaps between the hinge body and the hinge arm during relative rotation.\",\n \"Preferably, at least one abutment portion comprises a ridge or step upstanding from said body portion.\",\n \"Preferably, an abutment portion is provided on at least two of an inner side, an outer side and a bottom of at least one of the hinge body or hinge arm.\",\n \"Preferably, the hinge body includes an attachment portion for attachment of a cover for a working platform, and in use, said attachment portion serves to help secure the cover.\",\n \"Preferably, said attachment portion is adapted to pass through an aperture in a cover.\",\n \"Preferably, at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform.\",\n \"Preferably, at least one of said hinge arms includes an attachment portion in order to help secure a cover for a working platform and said attachment portion comprises a guide portion to guide an edge portion of said cover between an attachment point on a working platform and the attachment portion provided on the hinge body.\",\n \"Preferably, the first and second axes of rotation are substantially horizontal in use and respectively pass through the vertical thicknesses of the first and second hinge arms.\",\n \"Preferably, when the hinge assembly is in the open position, longitudinal axes of the first and second hinge arms are substantially co-axial with a longitudinal axis of the hinge body.\",\n \"Preferably, when the hinge assembly is in the closed-position respective longitudinal axes of the first and second hinge arms are substantially parallel, and the longitudinal axes of the first and second hinge arms are substantially perpendicular to a longitudinal axis of the hinge body.\",\n \"Further aspects of the present invention provide a working platform comprising first and second leaves mutually connected by one or more hinge assemblies in accordance with the first aspect.\",\n \"A further aspect of the present invention provides an ironing board comprising first and second leaves mutually connected by one or more hinge assemblies in accordance with the first aspect.\",\n \"In a particularly preferred embodiment of the present invention, a pair of hinge assemblies connect two leaves of the working platform of a folding ironing board.\",\n \"Preferably, the ironing board can be folded approximately centrally of the longitudinal extent of its working platform.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS Preferred embodiments of the present invention will now be described, by way of example only, with reference to the accompanying drawings, in which: FIG.\",\n \"1 is a perspective view of a hinge assembly in a fully open position; FIGS.\",\n \"2 to 4 are perspective views of the hinge assembly of FIG.\",\n \"1 in sequential intermediate positions in movement between a fully open position and a fully closed position; FIG.\",\n \"5 is a perspective view of the hinge assembly of FIG.\",\n \"1 in the fully closed position; FIG.\",\n \"6 is a partial perspective view illustrating the hinge assembly of FIG.\",\n \"1 in the fully open position and supporting a leaf of a two-piece working platform of a folding ironing board; FIGS.\",\n \"7 to 9 are partial perspective views of the hinge assembly and working platform of FIG.\",\n \"6 in sequential intermediate positions in movement between the fully open position and the fully closed position; FIG.\",\n \"10 is a partial perspective view of the hinge assembly and working platform of FIG.\",\n \"6 in the fully closed position; FIGS.\",\n \"11 to 13 are perspective views of the hinge assembly of FIG.\",\n \"1 in positions equivalent to those shown in FIGS.\",\n \"1, 3 and 5 respectively, illustrating the interaction in use of the hinge assembly with a cloth ironing board cover, during movement between the fully open position and the fully closed position, with the working platform omitted for simplicity; FIGS.\",\n \"14a and 14b are perspective views of a variation of a hinge arm; FIGS.\",\n \"14c, 14d, 14e and 14f are respectively outer side, bottom, top and end views of the hinge arm of FIGS.\",\n \"14a and 14b; FIGS.\",\n \"14g, 14h and 14i are cross-sectional views corresponding to the section lines G-G, H-H and I-I of FIG.\",\n \"14d; FIGS.\",\n \"15a and 15b are perspective views of a hinge body for use in the embodiment of FIG.\",\n \"1; FIGS.\",\n \"15c, 15d, 15e, 15f and 15g are outer side, bottom, top, inner side, and end views of the hinge body of FIGS.\",\n \"15a and 15b; and FIG.\",\n \"15h is a cross-sectional view on H-H of FIG.\",\n \"15e.\",\n \"DETAILED DESCRIPTION OF THE DRAWINGS Referring firstly to FIG.\",\n \"1, the principal elements of a hinge assembly 20 are a hinge body 22, a first hinge arm 24 and a second hinge arm 26.Hinge arms 24 and 26 are pivotally connected to hinge body 22 by first and second hinge pins 28, 30, respectively.\",\n \"The hinge pins 28 and 30 are spaced apart from one another and substantially parallel.\",\n \"The respective axes of rotation of the first and second hinge arms 24, 26 relative to the hinge body 22 are thus non-coaxial.\",\n \"As illustrated in FIG.\",\n \"1, the hinge assembly 20 is in a fully open position with the hinge arms 24, 26 extending horizontally away from the hinge body 22 in substantially opposing directions and being separated from one another by an angle of 180°.\",\n \"The first hinge arm 24 may be attached to a first leaf of a working platform, and the second hinge arm 26 may be attached to a second leaf of the working platform.\",\n \"As illustrated in the drawings, the first leaf could be supported by the second leaf via the hinge assembly (or a plurality of hinge assemblies) and the second leaf would be supported by external support elements such as frame, leg or strut elements.\",\n \"In movement from the open position illustrated in FIG.\",\n \"1 to the closed position illustrated in FIG.\",\n \"5, elements of the hinge assembly 20 rotate relative to each other first about the first hinge pin 28 and subsequently about the second hinge pin 30.Urging the distal end of the first hinge arm 24 upwardly results in its rotation relative to hinge body 22 about the first hinge pin 28 through the position illustrated in FIG.\",\n \"2.Continued rotation of the first hinge arm 24 results in closed-position mating surfaces 32 of the first hinge arm 24 meeting closed-position mating surfaces 34 of the hinge body 22 at junction 36 at the position illustrated in FIG.\",\n \"3.At this position, it can be seen that the first hinge arm 24 is generally perpendicular to the hinge body 22.Further rotation about hinge pin 28 is inhibited by the engagement of the closed-position mating surfaces 32, 36 and continued urging of the first hinge arm 24 causes the hinge body 22 to rotate relative to the second hinge arm 26, about the second hinge pin 30, through the position illustrated in FIG.\",\n \"4.Continued rotation of hinge body 22 about the second hinge pin 30 results in closed-position mating surfaces 38 of the hinge body 22 meeting closed-position mating surfaces 40 of the second hinge arm 26 at junction 42 at the closed position of the hinge assembly illustrated in FIG.\",\n \"5.In the closed position, the hinge arms 24 and 26 extend horizontally away from the hinge body 22 substantially parallel to one another, and substantially perpendicular to the hinge body 22.This illustrated order of relative movement is preferred, but not essential, and may vary, even in one embodiment due to factors such as wear or lubrication of different parts of the hinge assembly.\",\n \"Referring now to FIG.\",\n \"6, the first and second hinge arms 24, 26 are arranged to be received in respective first and second channels 44, 46 of an ironing board working platform 48 and secured thereto by locking tabs 50 which pop into complimentary slots or apertures in the channels 44 and 46.The working platform 48 is formed of first and second leaves 52, 54 which are secured together by a pair of hinge assemblies 20 (only one illustrated).\",\n \"The leaves 52 and 54 are illustrated in a working (open) position in FIG.\",\n \"6.By urging the first leaf 52, and by the consequent action of the hinge assemblies 20, the first leaf 52 is rotated relative to the second leaf through the positions illustrated in FIGS.\",\n \"7 to 9 until the hinge assembly is in its closed position and the first leaf 52 is in a storage position, illustrated in FIG.\",\n \"10.Only the second leaf 54 of working platform 48 is supported by external support elements 55.In the working position, illustrated in FIG.\",\n \"6, the first leaf 52 is supported solely by the second leaf via the hinge assemblies 20.In the working (open) position, the first leaf 52 is therefore supported in a cantilevered configuration.\",\n \"Referring back to FIGS.\",\n \"1 to 5, the support is provided by interaction between two pairs of open-position mating surfaces.\",\n \"A first pair of open-position mating surfaces comprises open-position mating surfaces 56 of the first hinge arm 24 and open-position mating surfaces 58 of the hinge body 22.A second pair of open-position mating surfaces comprises open-position mating surfaces 60 of the second hinge arm 26 and open-position mating surfaces 62 of the hinge body 22.The open-position mating surfaces 56, 58, 60, 62 of each pair abut when the hinge assembly is in the open position, illustrated in FIG.\",\n \"1.The interaction between the mating surfaces 56, 58, 60, 62 is sufficient to support a working load on the first leaf 52 in the working position of FIG.\",\n \"6.The edges 64 and 66 of leaves 52 and 54 respectively are formed with curved lips 68 and 70 respectively which may be guided by leaf guide 72 of hinge body 22 during movement between the positions illustrated in FIGS.\",\n \"6 and 10, to maintain the pair of hinge assemblies 20 rotating in unison and thereby minimise the tendency of the edges 64 and 66 to twist relative to one another.\",\n \"In use, the leaves 52 and 54 will typically be covered by a thin sheet of foam (not shown) and a material cover 74 (see FIGS.\",\n \"11-14).\",\n \"Each hinge body 22 is formed with a cover attachment portion in the form of button 76 which retains the material cover 74 by a button hole 78 which maintains the cover 74 taut in the operating position of FIG.\",\n \"11 and prevents the cover 74 coming away from leaves 52 and 54 in the storage position, illustrated in FIG.\",\n \"13.Each hinge arm 24 and 26 is formed with a guide tab 80 which guides movement of an elasticised edge 82 of the material cover 74 during rotation of the hinge assemblies 20.The main elements of the hinge assembly will now be described in more detail.\",\n \"With reference to FIGS.\",\n \"14a to 14i, the illustrated hinge arm corresponds to, but is slightly different to the second hinge arm 26 of FIGS.\",\n \"1 to 13 since it is not provided with locking tabs (50 in FIG.\",\n \"1).\",\n \"However, since the embodiment of FIGS.\",\n \"14a to 14i has many common features to that of the embodiment of FIGS.\",\n \"1 to 13, corresponding reference numerals will be used for corresponding features.\",\n \"The hinge arm 26 of FIGS.\",\n \"14a to 14i has an elongate fitting portion 200 for fitting into a channel of an ironing board working platform, and a connection portion 250 for connection and engagement with the hinge body 22 of a hinge assembly 20.The fitting portion 200 has a rearmost end distal from the connection portion 250 and a foremost end connected to the connection portion 250.The fitting portion 200 is generally I-shaped in cross-section, as best shown in FIG.\",\n \"14h.\",\n \"The hinge arm 26 may be considered to have an outer side, which in use, faces away from the working platform, and an inner side which, in use, faces towards the centre of the working platform.\",\n \"On the outer side of the connection portion 250 is located the guide tab 80 which provides a generally downwardly facing guide surface 280 in order to guide movement of an elasticised edge of a material cover (shown in FIGS.\",\n \"11 to 13).\",\n \"The guide surfaces are inclined and run in a direction which is generally aligned from the bottom (or non-working surface) extreme of the hinge arm towards the button.\",\n \"The connection portion 250 includes a cavity 252 for receiving an end portion of a hinge body 22, the cavity 252 being defined by opposing side wall portions 254, 256.Each of the opposing wall portions 254, 256 has a respective circular aperture 255, 257 therein for receiving a hinge pin 30 in order to pivotally attach the hinge arm 26 to the hinge body 22.The opposing wall portions 254, 256 have respective front edges 260, 262 which are arcuate and which have a centre of curvature located at the centre of the respective apertures 255, 257.In use, the front edges 260, 262 run close to or bear against corresponding surfaces of the hinge body 22 (described in detail hereafter) in a configuration which helps avoid catching of objects between the hinge arm 26 and the hinge body 22 during operation, and which may provide support for the hinge pins if required.\",\n \"The opposing wall portions 254, 256 are connected together by a bottom wall portion 264 and a top wall portion 266.As shown in FIG.\",\n \"14c, the bottom wall portion 264 extends forwardly from the foremost end of the fitting portion 200 to a position directly beneath the centre of the apertures 255, 257.As is best shown in FIGS.\",\n \"14a, 14b, 14f and 14g the bottom wall is arched downwardly at its foremost part providing an arcuate inner surface 265.The arcuate foremost edge of the bottom wall 264 provides the open-position mating surfaces 60 of the second hinge arm 26.The top wall 266 is relatively short, extending forwardly from the foremost end of the fitting portion 200 approximately half as far as the bottom wall 264 extends.\",\n \"The top edges of the wall portions 254, 256 provide the closed-position mating surfaces 38 of the second hinge arm.\",\n \"(A foremost edge 268 of the top wall 266 may also provide a closed-position mating surface 38 of the second hinge arm, by engagement with the top surface of the hinge body 22, if required.)\",\n \"The preferred embodiment of a hinge arm is made from aluminium and may conveniently be formed from two separately formed longitudinal “halves”, so that the finished hinge arm has a longitudinal joint line along its length, approximately half way between the inner and outer sides thereof.\",\n \"Although only one hinge arm 26 is described in detail, and illustrated in FIG.\",\n \"14, it will be appreciated that the first hinge arm 24 corresponds, but is a mirror image thereof.\",\n \"With reference to FIGS.\",\n \"15a to 15h, the preferred embodiment of the hinge body 22 will now be described in detail.\",\n \"The hinge body 22 includes first and second hinge body end portions 302, 304 at respective first and second ends thereof.\",\n \"Each end portion 302, 304 is laterally narrow compared to the rest of the hinge body 22 and includes a respective circular aperture 306, 308 therethrough for receiving a respective hinge pin 28, 30 and thereby connecting the hinge body 22 to respective hinge arms 24, 26.Each end portion 302, 304 has a respective arcuate edge portion 310, 312, being approximately quarter circular, extending from an endmost edge part of the respective end portion to a bottommost edge of the respective end portion, and having the respective apertures 306, 308 as their respective centres of curvature.\",\n \"The arcuate edge portions 310, 312 have rounded surfaces in the lateral direction of the body end portions 302, 304, so that they can slide neatly into and out of the spaces defined by the arcuate inner surfaces 265 of the hinge arms 24, 26, when the hinge body 22 and hinge arms 24, 26 are rotated relative to each other.\",\n \"This sliding action avoids the creation and termination of gaps, in which objects could get caught, during operation of the hinge assembly.\",\n \"Intermediate the two hinge body end portions 302, 304 there is a central body portion 314 which has a first side projecting portion 314a projecting to an outer lateral side of the hinge body 22 and a second side projecting portion 314b projecting to an inner lateral side of the hinge body 22.The first and second side portions 314a, 314b of the central body portion 314 meet at the bottom of the hinge body 22 so that the central body portion 314 effectively wraps around the sides and bottom of the hinge body 22.The projecting central body portion 314 provides a number of step or ridge portions which form narrow mating or bearing surfaces for interaction with surfaces on the hinge arms 24, 26.In this embodiment, the mating or bearing surfaces provided project substantially perpendicular to the adjacent surfaces of the end portions 302, 304 of the hinge body 22.Because the projecting central body portion 314 extends substantially symmetrically about both sides of the hinge body 22, similar mating or bearing surfaces are provided on the inner and outer sides of the hinge body 22.For convenience only those provided on the outer side will be described in detail below, and the reference numerals used will be suffixed by the letter “a”.\",\n \"Corresponding mating surfaces provided on the inner side of the hinge body 22 are designated in the drawings by the same reference numerals but with the suffix “b” replacing the suffix “a”.\",\n \"Towards the top of the hinge body (as shown, for example in FIG.\",\n \"15a) the central body portion is relatively small in longitudinal extent and provides substantially straight, substantially parallel, substantially vertical closed-position mating surfaces 34a, 38a, which are adapted to abut mating surfaces 32, 40 of the hinge arms formed by the top edges of the side wall portions 254, 256.It is noted for clarity that directional descriptors to the top, bottom and the like of certain elements are used for convenience despite the rotation of those elements in a vertical plane, in use.\",\n \"Such directional descriptors herein refer to the relative directions when the elements are in the orientation shown in FIGS.\",\n \"1, 6 and 11, that is with the hinge assembly 20 fully open and the working platform in a substantially horizontal configuration, and should be construed accordingly.\",\n \"The projecting central body portion 314 increases in longitudinal extent as it extends downwards (from the bottom of the closed-position mating surfaces 34a, 38a) to provide arcuate bearing surfaces 316a, 318a which have centres of curvature corresponding to the centres of the apertures 306, 308.The arcuate bearing surfaces 316a, 318a extend from the bottom of the vertical mating surfaces 34a, 38a through a quarter circle to respective points substantially vertically below the centres of the apertures 306, 308.These arcuate bearing surfaces 316a, 318a (and corresponding 316b, 318b on the inner side of the hinge body 22) are adapted in use to slide against, or close to, the arcuate edges 260, 262 of the hinge arms, as discussed above.\",\n \"Also formed by the central body portion 314 where it passes around the underside of the hinge body 22 and extends between the inner and outer sides of the hinge body 22 are open-position mating surfaces 58, 62.These take the form, as best illustrated in FIGS.\",\n \"15a, 15b and 15g, of vertically orientated semi-annular surfaces.\",\n \"In use, these open-position mating surfaces 58, 62 are adapted to abut the open-position mating surfaces 56, 60 provided on the hinge arms 24, 26.On the outer side of the central body portion 314a is provided the button 76.On the inner side of the central body portion 314b is provided the leaf guide 72 which provides first and second arcuate guide surfaces 320, 322 adapted to bear against edges of the two leaves.\",\n \"The leaf guide 72 is provided in order to guide respective leaves of a working platform and help prevent relative twisting of the two leaves which, in practice, would normally be connected by a pair of hinge assemblies 20.The hinge body 22 is preferably die cast from aluminium.\",\n \"It will be appreciated that the hinge assembly 20 described has a number of beneficial features.\",\n \"It allows a working platform to be provided in which only one of the leaves needs to be supported by external support elements, since the externally supported leaf can support the other leaf in cantilevered configuration via the hinge assembly or assemblies 20.This provides convenient compact support, and great ease of use as no deployment of external support elements is required.\",\n \"It is normally preferred to use a pair of hinge assemblies to connect two leaves but in certain applications, for example where the working platform is relatively narrow and provided with a central channel, use of a single hinge assembly might be appropriate.\",\n \"In the fully open position, the described hinge assembly fits entirely within the thickness (or height) of the hinge arms 24, 26 and this is advantageous over the use of hinge assemblies which protrude above or below the hinge arms and the platform.\",\n \"When the hinge assembly is in its closed position, the only part of the hinge assembly exposed at the folded terminus of the working platform is the bottom surface of the hinge body 22, providing a blunt and safe platform end, unlike some other hinge assemblies.\",\n \"Rotation of the hinge arms about two non-coaxial axes of rotation allows first and second leaves to be stored parallel and adjacent each other.\",\n \"Different thicknesses of working platform or working platform cover (for example, foam padding and/or an ironing board cover) can be accommodated by use of different lengths of hinge body 22.The use of snug-fitting arcuate sliding portions on the hinge body 22 and hinge arms 24, 26 provides additional support to the structure (if this is required due to wear and or play in the hinge pins 28, 30) and minimises the likelihood of items being caught in gaps which vary in size as the hinge assembly is operated.\",\n \"The provision of closed-position mating surfaces 32, 34, 38, 40 to keep the hinge arms (or leaves) substantially parallel when the, hinge assemblies are closed reduces the likelihood of items, such as fingers, being damaged by being caught between the leaves as the platform is folded.\",\n \"Although desirable these may not always be necessary, as under some circumstances it may be adequate to allow the leaves of the working surface to contact each other in the closed position.\",\n \"The provision of a button 76 or other suitable cover retaining means on the hinge body 22 helps to retain the cover in position during folding and unfolding.\",\n \"Provision of guide tabs 80 further help to retain the cover and prevent it from becoming caught in the hinge assembly.\",\n \"Throughout this specification and the claims, the words “comprise”, “comprises” and “comprising” are used in a non-exclusive sense, except where the context requires otherwise.\",\n \"It is to be understood that, if any prior art publication is referred to herein, such reference does not constitute an admission that the publication forms a part of the common general knowledge in the art, in Australia or in any other country.\",\n \"Modifications and improvements may be incorporated without departing from the scope of the present invention.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476907"}}},{"rowIdx":981,"cells":{"text":{"kind":"list like","value":[["Trailing beam suspension with alignment adjustment","A trailing beam suspension includes a mechanical adjustment mechanism for aligning the axle and wheels of a vehicle.","The trailing beam is mounted to the vehicle frame by a pivot bolt passing through a hanger bracket and a conventional bushed connection.","Opposing sides of the hanger bracket contain aligned curved apertures that receive the pivot bolt.","The pivot bolt passes through pivot arms which are pivotally connected to the sides of the hanger bracket.","The pivot arms are connected to a yoke to which an adjustment shaft is attached.","Movement of the adjustment shaft moves the yoke fore and aft.","The movement of the yoke pivots the pivot arms, thus moving the pivot bolt in a fore-and aft-direction.","When the trailing beam has been aligned, the pivot connection is tightened, thus securing the trailing beam, axle, and wheels in the desired alignment."],["1.In a trailing beam suspension for a vehicle comprising a hanger bracket adapted to be mounted to a vehicle frame rail, a trailing beam pivotally mounted at one end to the hanger bracket through a pivot pin and an adjustment slot in the hanger bracket so that the position of the beam with respect to the hanger bracket is selectively adjustable in a fore-and-aft direction to align an axle mounted to the trailing beam, the improvement comprising: a manually adjustable, threadably actuated drive mechanism mounted between the hanger bracket and the pivot bolt for incrementally adjusting the position of the pivot bolt in the slot to thereby adjust the beam with respect to the hanger bracket, wherein the drive mechanism includes a drive shaft that is substantially orthogonal to an axis of the pivot bolt and is substantially offset therefrom.","2.The trailing beam suspension according to claim 1 wherein the pivot pin is mounted to at least one pivot arm, the pivot arm is pivotally mounted at one end to the hanger bracket for pivotal movement about an axis spaced from the adjustment slot and the drive mechanism is mounted to the at least one pivot arm at a location spaced from the adjustment slot and from the axis of the pivot arm, whereby pivotal movement of the at least one pivot arm selectively adjusts the relative position of the pivot bolt in the adjustment slot.","3.The trailing beam suspension according to claim 2 wherein the drive mechanism further comprises a drive shaft connected to the at least one pivot arm to adjustably move the pivot arm as the shaft is driven.","4.The trailing beam suspension according to claim 3 wherein the drive shaft is journaled in the hanger bracket and is threadably mounted to the at least one pivot arm.","5.In a trailing beam suspension for a vehicle comprising a hanger bracket adapted to be mounted to a vehicle frame rail, a trailing beam pivotally mounted at one end to the hanger bracket through a pivot pin and an adjustment slot in the hanger bracket so that the position of the beam with respect to the hanger bracket is selectively adjustable in a fore-and-aft direction to align an axle mounted to the trailing beam, the improvement comprising: a drive mechanism mounted between the hanger bracket and the pivot bolt for incrementally adjusting the position of the pivot bolt in the slot to thereby adjust the beam with respect to the hanger bracket, wherein the pivot pin is mounted to at least one pivot arm, the pivot arm is pivotally mounted at one end to the hanger bracket for pivotal movement about an axis spaced from the adjustment slot and the drive mechanism is mounted to the at least one pivot arm at a location spaced from the adjustment slot and from the axis of the pivot arm, whereby pivotal movement of the at least one pivot arm selectively adjusts the relative position of the pivot bolt in the adjustment slot, the drive mechanism further comprising a drive shaft connected to the at least one pivot arm to adjustable move the pivot arm as the shaft is driven, and wherein the drive shaft being journaled in the hanger bracket and is threadably mounted to the at least one pivot arm and mounted to the at least one pivot arm through a yoke that has a threaded aperture in which the drive shaft is received.","6.The trailing beam suspension according to claim 5 wherein the longitudinal axis of the shaft is orthogonal to the axis of the at least one pivot arm.","7.The trailing beam suspension according to claim 6 wherein the hanger bracket has a pair of spaced side walls and an end wall joining the side walls, there are two pivot arms, one adjacent to each side wall of the hanger bracket and the drive shaft is journaled in the end wall.","8.The trailing beam suspension according to claim 1 wherein the drive mechanism includes a threaded shaft.","9.The trailing beam suspension according to claim 8 wherein rotation of the threaded shaft moves the pivot bolt in the slot.","10.The trailing beam suspension according to claim 1 wherein the drive mechanism includes a linear hydraulic actuator.","11.The trailing beam suspension according to claim 1 wherein the drive mechanism includes a linear pneumatic actuator.","12.The trailing beam suspension according to claim 1 wherein the drive mechanism includes an electrically-powered actuator.","13.The trailing beam suspension according to claim 7 wherein the shaft is substantially centrally located between the side walls of the hanger bracket.","14.The trailing beam suspension according to claim 1 wherein the drive mechanism symmetrically engages the pivot bolt."],["<SOH> BACKGROUND OF THE INVENTION <EOH>1.Technical Field The invention relates to vehicle suspensions.","In one aspect, the invention relates to a trailing beam suspension for a wheeled vehicle in which the suspension includes a mechanism for fore or aft adjustment of the alignment of the axle and wheels with respect to the longitudinal axis of the vehicle.","2.Related Art Proper wheel alignment in road vehicles is important to prevent undue tire wear.","Common causes of improper wheel alignment include improper initial suspension system installation, accident damage, and impact or shock loads imposed on suspension components by potholes or other irregularities in the road.","Wheel alignment adjustment mechanisms permit adjustment of the alignment of vehicle axles and wheels with respect to the vehicle frame.","Adjustment mechanisms are generally one of two types: (1) those that are adjustable as the suspension is installed on the vehicle and then permanently fixed in the adjusted position by welding or the like, and (2) those that are adjustable any time during and after installation and are held in the adjusted position mechanically.","U.S. Pat.","No.","3,960,388 to Strader et al., which is incorporated herein in its entirety by reference, is an example of the first category of alignment devices.","The Strader '388 patent discloses a trailing beam suspension that incorporates a mounting plate for the trailing beam having a pair of bosses that engage slots in the hanger bracket.","The trailing beam is attached to the hanger bracket by a bolted connection in which the bolt passes through the mounting plate and is secured with a nut.","The bolt head engages the mounting plate so that rotating the bolt pivots the mounting plate about one of the bosses.","Pivoting of the mounting plate moves the bolt fore or aft, which moves the trailing beam fore or aft to align the axle.","When the axle and wheels have been properly aligned, the plate is welded to the hanger bracket.","Further adjustment is possible only after removing the weld.","U.S. Pat.","No.","Re.","30,308 to Masser discloses a trailing beam suspension system incorporating a mounting plate for the trailing beam having a single boss that engages a slot in the hanger bracket.","The pivot bolt for the trailing beam passes through the mounting plate and slot.","The trailing beam is moved fore or aft until proper alignment of the axle and wheels is achieved.","The mounting plate is then welded to the hanger bracket.","Further adjustment is possible only after removing the weld.","U.S. Pat.","No.","4,991,872 to Richardson discloses a trailing beam suspension for a low-bed trailer.","The forward trailing beam of a pair of trailing beams is mounted to the intermediate, inclined portion of the trailer's main structural beam through a bushing assembly.","The bushing assembly is held to the web of the beam by a bolted connection.","The web is provided with slotted mounting holes that enable adjustment of the alignment of the trailing beam and attached axle.","The trailing beam is moved fore or aft until the axle and wheels are properly aligned.","After adjustment, the bushing assembly is welded to the beam.","Further adjustment requires removal of the weld and loosening of four bolts connecting the bushing assembly to the trailer beam.","An example of the second category of alignment devices is disclosed in U.S. Pat.","No.","5,201,898 to Pierce.","The Pierce '898 patent discloses a trailing beam suspension incorporating a pair of complementary discs that engage each other with a channel and rib arrangement, with the trailing beam mounting bolt passing therethrough.","One disc is welded to the hanger bracket and incorporates an asymmetrical hole.","The second disc contains an off-center hole and is rotatable relative to the first.","The rotation of the second disc relative to the first is translated into lateral movement of the trailing beam mounting bolt.","It is necessary to restrain the second disc against further rotation while the connection is tightened to prevent movement of the trailing beam from its aligned position."],["<SOH> SUMMARY OF THE INVENTION <EOH>According to the invention, a trailing beam suspension for a vehicle comprises a hanger bracket adapted to be mounted to a vehicle frame rail, a trailing beam pivotally mounted at one end to the hanger bracket through a pivot pin and an adjustment slot in the hanger bracket so that the position of the beam with respect to the hanger bracket is selectively adjustable in a fore-and-aft direction to align an axle mounted to the trailing beam.","According to the invention, a drive mechanism is mounted between the hanger bracket and the pivot bolt for incrementally adjusting the position of the pivot bolt in the slot to thereby adjust the beam with respect to the hanger bracket.","In one embodiment, the pivot pin is mounted to at least one pivot arm, the pivot arm is pivotally mounted at one end to the hanger bracket for pivotal movement about an axis spaced from the adjustment slot and the drive mechanism is mounted to the at least one pivot arm at a location spaced from the adjustment slot and from the axis of the pivot arm.","Pivotal movement of the at least one pivot arm selectively adjusts the relative position of the pivot bolt in the adjustment slot.","Preferably, the drive mechanism further comprises a drive shaft connected to the at least one pivot arm to adjustably move the pivot arm as the shaft is driven.","In one embodiment, the drive shaft is journaled in the hanger bracket and is threadably mounted to the at least one pivot arm.","Further, in a preferred embodiment, the drive shaft is mounted to the at least one pivot arm through a yoke that has a threaded aperture in which the drive shaft is received.","Typically, the longitudinal axis of the shaft is orthogonal to the axis of the at least one pivot arm.","In a preferred embodiment, the hanger bracket has a pair of spaced side walls and an end wall joining the side walls.","There are two pivot arms, one adjacent to each side wall of the hanger bracket and the drive shaft is journaled in the end wall of the hanger bracket.","In a preferred embodiment of the invention, the drive mechanism includes a threaded shaft and rotation of the threaded shaft moves the pivot bolt in the slot.","In another embodiment, the drive mechanism includes a linear hydraulic actuator.","In still another embodiment, the drive mechanism includes a linear pneumatic actuator.","In yet another alternative embodiment, the drive mechanism includes an electrically-powered actuator."],["CROSS-REFERENCE TO RELATED APPLICATION This application claims the benefit of U.S.","Provisional Patent Application Ser.","No.","60/288,935, filed on May 4, 2001.BACKGROUND OF THE INVENTION 1.Technical Field The invention relates to vehicle suspensions.","In one aspect, the invention relates to a trailing beam suspension for a wheeled vehicle in which the suspension includes a mechanism for fore or aft adjustment of the alignment of the axle and wheels with respect to the longitudinal axis of the vehicle.","2.Related Art Proper wheel alignment in road vehicles is important to prevent undue tire wear.","Common causes of improper wheel alignment include improper initial suspension system installation, accident damage, and impact or shock loads imposed on suspension components by potholes or other irregularities in the road.","Wheel alignment adjustment mechanisms permit adjustment of the alignment of vehicle axles and wheels with respect to the vehicle frame.","Adjustment mechanisms are generally one of two types: (1) those that are adjustable as the suspension is installed on the vehicle and then permanently fixed in the adjusted position by welding or the like, and (2) those that are adjustable any time during and after installation and are held in the adjusted position mechanically.","U.S. Pat.","No.","3,960,388 to Strader et al., which is incorporated herein in its entirety by reference, is an example of the first category of alignment devices.","The Strader '388 patent discloses a trailing beam suspension that incorporates a mounting plate for the trailing beam having a pair of bosses that engage slots in the hanger bracket.","The trailing beam is attached to the hanger bracket by a bolted connection in which the bolt passes through the mounting plate and is secured with a nut.","The bolt head engages the mounting plate so that rotating the bolt pivots the mounting plate about one of the bosses.","Pivoting of the mounting plate moves the bolt fore or aft, which moves the trailing beam fore or aft to align the axle.","When the axle and wheels have been properly aligned, the plate is welded to the hanger bracket.","Further adjustment is possible only after removing the weld.","U.S. Pat.","No.","Re.","30,308 to Masser discloses a trailing beam suspension system incorporating a mounting plate for the trailing beam having a single boss that engages a slot in the hanger bracket.","The pivot bolt for the trailing beam passes through the mounting plate and slot.","The trailing beam is moved fore or aft until proper alignment of the axle and wheels is achieved.","The mounting plate is then welded to the hanger bracket.","Further adjustment is possible only after removing the weld.","U.S. Pat.","No.","4,991,872 to Richardson discloses a trailing beam suspension for a low-bed trailer.","The forward trailing beam of a pair of trailing beams is mounted to the intermediate, inclined portion of the trailer's main structural beam through a bushing assembly.","The bushing assembly is held to the web of the beam by a bolted connection.","The web is provided with slotted mounting holes that enable adjustment of the alignment of the trailing beam and attached axle.","The trailing beam is moved fore or aft until the axle and wheels are properly aligned.","After adjustment, the bushing assembly is welded to the beam.","Further adjustment requires removal of the weld and loosening of four bolts connecting the bushing assembly to the trailer beam.","An example of the second category of alignment devices is disclosed in U.S. Pat.","No.","5,201,898 to Pierce.","The Pierce '898 patent discloses a trailing beam suspension incorporating a pair of complementary discs that engage each other with a channel and rib arrangement, with the trailing beam mounting bolt passing therethrough.","One disc is welded to the hanger bracket and incorporates an asymmetrical hole.","The second disc contains an off-center hole and is rotatable relative to the first.","The rotation of the second disc relative to the first is translated into lateral movement of the trailing beam mounting bolt.","It is necessary to restrain the second disc against further rotation while the connection is tightened to prevent movement of the trailing beam from its aligned position.","SUMMARY OF THE INVENTION According to the invention, a trailing beam suspension for a vehicle comprises a hanger bracket adapted to be mounted to a vehicle frame rail, a trailing beam pivotally mounted at one end to the hanger bracket through a pivot pin and an adjustment slot in the hanger bracket so that the position of the beam with respect to the hanger bracket is selectively adjustable in a fore-and-aft direction to align an axle mounted to the trailing beam.","According to the invention, a drive mechanism is mounted between the hanger bracket and the pivot bolt for incrementally adjusting the position of the pivot bolt in the slot to thereby adjust the beam with respect to the hanger bracket.","In one embodiment, the pivot pin is mounted to at least one pivot arm, the pivot arm is pivotally mounted at one end to the hanger bracket for pivotal movement about an axis spaced from the adjustment slot and the drive mechanism is mounted to the at least one pivot arm at a location spaced from the adjustment slot and from the axis of the pivot arm.","Pivotal movement of the at least one pivot arm selectively adjusts the relative position of the pivot bolt in the adjustment slot.","Preferably, the drive mechanism further comprises a drive shaft connected to the at least one pivot arm to adjustably move the pivot arm as the shaft is driven.","In one embodiment, the drive shaft is journaled in the hanger bracket and is threadably mounted to the at least one pivot arm.","Further, in a preferred embodiment, the drive shaft is mounted to the at least one pivot arm through a yoke that has a threaded aperture in which the drive shaft is received.","Typically, the longitudinal axis of the shaft is orthogonal to the axis of the at least one pivot arm.","In a preferred embodiment, the hanger bracket has a pair of spaced side walls and an end wall joining the side walls.","There are two pivot arms, one adjacent to each side wall of the hanger bracket and the drive shaft is journaled in the end wall of the hanger bracket.","In a preferred embodiment of the invention, the drive mechanism includes a threaded shaft and rotation of the threaded shaft moves the pivot bolt in the slot.","In another embodiment, the drive mechanism includes a linear hydraulic actuator.","In still another embodiment, the drive mechanism includes a linear pneumatic actuator.","In yet another alternative embodiment, the drive mechanism includes an electrically-powered actuator.","BRIEF DESCRIPTION OF THE DRAWINGS The invention will be described with reference to the accompanying drawings in which: FIG.","1 is a side elevational view of a trailing beam suspension incorporating the present invention.","FIG.","2 is a top perspective view of a preferred embodiment of the trailing beam suspension of FIG.","1 showing a yoke assembly installed in a hanger bracket and a trailing beam pivot connection.","FIG.","3 is a perspective view of the hanger bracket illustrated in FIG.","2.FIG.","4 is a perspective view of a portion of the yoke assembly illustrated in FIG.","2.FIG.","5 is a perspective view of a pivot arm illustrated in FIG.","2.FIG.","6 is a side perspective view of a subassembly consisting of the yoke assembly, pivot arms, and pivot bolt illustrated in FIG.","2.FIG.","7 is a side view of the subassembly of FIG.","6 installed in the hanger bracket and illustrating the relative movement of the components of the alignment mechanism as the device is operated.","FIG.","8 is a perspective view of a hanger bracket comprising part of an alternate embodiment of the trailing beam suspension of FIG.","1.FIG.","9 is a perspective view of a pivot arm which is complementary to the hanger bracket illustrated in FIG.","8.DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Referring now to the drawings, and in particular to FIG.","1, there is illustrated a trailing beam suspension system for a vehicle generally identified by the numeral 10.The suspension system comprises a trailing beam 20, a hanger bracket 22, and an air spring 30.The hanger bracket 22 is attached in a conventional manner to a vehicle frame rail 12.A first end of the trailing beam 20 is pivotally mounted to the hanger bracket 22 through a pivot bolt 26 and a conventional bushed connection (not shown) in the same manner as the trailing arm 22 is pivotally mounted to the bolt 34 in the Strader et al.","U.S. Pat.","No.","3,960,388.A second end of the trailing beam 20 is attached in a conventional manner to the air spring 30 so that the air spring 30 is disposed in load transmitting relation between the trailing beam 20 and the frame rail 12.An axle 16, with attached wheels 14, is connected to the trailing beam 20 by an axle bracket 18.Referring now to FIGS.","2-6, a preferred embodiment of an alignment mechanism generally comprises the hanger bracket 22, a pivot pin 25 comprising a threaded pivot bolt 26 with a complementary nut 44, an adjustable yoke assembly 39, and two pivot arms 46 with attached pivot studs 38.The hanger bracket has two parallel, laterally spaced-apart sides comprising a first side 56 and a second side 58 joined by an intermediate side 60 generally orthogonal thereto.","Washers 42 are spaced between the outer faces of the hanger bracket sides 56, 58, and the head of the pivot bolt 26 and nut 44, respectively.","Referring specifically to FIG.","3, the first and second sides 56, 58 of the hanger bracket 22 contain arcuate slots 62 in complementary alignment located at a lower portion of the hanger bracket 22.The arcuate slots 62 are elongate in a direction longitudinal of the trailing beam 20, and upwardly concave.","The first and second sides 56, 58 of the hanger bracket 22 also contain pivot apertures 64 in complementary alignment, which are located vertically beneath the arcuate slots 62 proximate to the lower edge of the hanger bracket 22.The arcuate slots 62 slidably receive the pivot bolt 26 and to allow the pivot bolt 26 to laterally translate therein.","The pivot apertures 64 slidably receive the pivot studs 38 and to allow rotation of the pivot studs 38 therein.","In the embodiment described herein, the upper portion of the intermediate side 60 contains an adjustment bolt aperture 52 and a slot 54 extending generally downwardly from the adjustment bolt aperture 52.The slot 54 accommodates the shaft of an adjustment bolt 28 to enable the adjustment bolt 28 to be freely turned therein.","FIGS.","2, 4 and 6 show the yoke assembly 39 of the preferred embodiment comprising the adjustment bolt 28 with a bolt head 29, a retainer nut 50, a yoke 40 having a threaded yoke aperture 72, and a yoke nut 48.As hereinafter described, turning of the adjustment bolt 28 in the yoke aperture 72 translates the yoke 40 to pivot the pivot arms 46 about the pivot studs 38.Alternate assemblies for urging the pivot arms 46 to pivot about the pivot studs 38 can be employed, such as a linear hydraulic, linear pneumatic, or electrically-powered actuator.","For example, a linear hydraulic actuator can have a hydraulic piston with a piston rod attached to the pivot arms 46 through an articulating connection so that extension and retraction of the hydraulic piston and piston rod pivots the pivot arms 46 fore and aft.","In the embodiment shown in FIGS.","2, 4 and 6, the retainer nut 50 is threaded onto the adjustment bolt 28 until it is proximate to the bolt head 29, leaving a gap between the retainer nut 50 and bolt head 29 that is slightly larger than the thickness of the intermediate side 60.The retainer nut 50 is fixedly attached to the adjustment bolt 28, such as by welding or brazing.","The yoke 40 comprises a hub 66, opposing arms 68, and a yoke aperture 72.The hub 66 is generally cylindrical and contains the yoke aperture 72 extending axially therethrough.","Extending radially from the hub 66 are the diametrically opposed arms 68.The arms 68 are generally cylindrical, axially aligned, and terminate in a pair of spaced-apart axially-opposed yoke studs 70.The yoke stud 70 is a cylindrical projection with a diameter that is smaller than the arm diameter and which is axially aligned with the arm 68.The length of the yoke stud 70 is slightly less than the thickness of the pivot arm 46.A recess 74 extends across the center portion of the hub 66 intermediate the arms 68.The recess 74 accommodates the yoke nut 48 to restrain the yoke nut 48 against rotation relative to the yoke 40.The yoke nut 48 is axially aligned with the yoke aperture 72 and is fixedly attached to the yoke 40, such as by welding or brazing.","The yoke 40 and yoke nut 48 can be threaded onto the adjustment bolt 28 by first passing the adjustment bolt 28 through the yoke aperture 72 and into the yoke nut 48.Referring now to FIG.","5, the pivot arm 46 is a generally plate-like elongated member with a central portion 84 that tapers to a first end 76 and to a second end 78.The central portion 84 contains a pivot bolt aperture 82 centered with respect to the longitudinal axis of the pivot arm 46.The first end 76 contains a shaft receptacle 80 centered with respect to the longitudinal axis of the pivot arm 46.The second end 78 has a pivot stud 38 extending orthogonally therefrom and centered with respect to the longitudinal axis of the pivot arm 46.The shaft receptacle 80 slidably receives the yoke stud 70.The pivot bolt aperture 82 slidably receives the pivot bolt 26.The pivot stud 38 is slidably received within the pivot aperture 64.The length of the pivot stud 38 is slightly greater than the thickness of a hanger bracket side 56, 58.Referring now to FIGS.","6 and 7, the assembly and operation of the invention will be described.","The adjustment bolt 28 is inserted through the yoke aperture and threaded into the yoke nut 48.The yoke studs 70 are inserted into the shaft receptacles 80 through the pivot arms 46, enabling each pivot arm 46 to pivot about its respective yoke stud 70.The pivot studs 38 are inserted into the pivot apertures 64 in the hanger bracket sides 56, 58, enabling the pivot arms 46 to pivot about the pivot studs 38.The adjustment bolt head 29 is passed through the adjustment bolt aperture 22, and the adjustment bolt 28 is positioned in the slot 54 so that the bolt head 29 is to the outside of the intermediate side 60 and the retainer nut 50 is to the inside of the intermediate side 60.The adjustment bolt 28 can be turned freely in the slot 54 and is restrained against axial movement by the bolt head 29 and retainer nut 50.Furthermore, the adjustment bolt 28 can move vertically within the slot 54 during the adjustment process.","The pivot bolt apertures 82 are also aligned with the arcuate slots 62.The pivot bolt 26 is inserted through a washer 42, a pivot bolt aperture 82, an arcuate slot 62, the bushed connection 24 of the trailing beam, the second arcuate slot 62, the second pivot bolt aperture 82, the second washer 42, and threaded into the nut 44 to pivotally connect the trailing beam to the hanger bracket 22.As assembled, each pivot arm 46 is in slidable contact with the inside face of its respective hanger bracket side 56, 58.After the alignment subassembly has been assembled in the hanger bracket 22 with the trailing beam 20 mounted to the pivot bolt 26 as described above, the alignment of the trailing beam is accomplished as follows.","The pivot bolt 26 and nut 44 are in a slightly loosened configuration.","The adjustment bolt 28 is rotated, which draws the yoke assembly 39 fore or aft, depending upon the direction of rotation of the adjustment bolt 28.Movement of the yoke assembly 39 pivots the pivot arms 46 about the pivot stud 38.This moves the pivot bolt 26 along the arcuate slots 62 in a corresponding fore or aft direction, which correspondingly moves the trailing beam 20 in a fore or aft direction.","This process is continued until the axle 16 and wheels 14 are properly aligned with the vehicle frame 12.When the axle 16 and wheels 14 are in the desired alignment, the nut 44 is tightened to secure the bushed connection 24 to the hanger bracket 22.The trailing beam 20, the axle 16, and the wheels 14 can be easily and accurately aligned due to the unique mechanism.","Additionally, the pivot bolt nut 44 can be securely tightened to the proper torque without the risk that the trailing beam 20 will become misaligned during the tightening process.","Tightening the pivot bolt nut 44 to the proper torque reduces the risk that the nut 44 will loosen over time and the pivot connection 24 will fail due to under-torquing of the nut 44.Additionally, the pivot bolt 26 is securely restrained against lateral movement in the slots 62 unless the adjustment bolt 28 is turned.","The unique alignment mechanism thus provides further protection against misalignment of the trailing beam 20, axle 16, and wheels 14 due to impact or shock loads imposed on the suspension by potholes or other road surface irregularities.","The trailing beam 20, axle 16, and wheels 14 will remain in the properly aligned position even if the nut 44 becomes loosened.","Furthermore, the alignment of the trailing beam 20 can be maintained without the necessity of welding the alignment components to the hanger bracket or vehicle frame, thus facilitating future adjustment to the wheel alignment.","An alternate embodiment of the invention comprises a hanger bracket with alternate arcuate slots and pivot apertures which is generally similar to the hanger bracket 22, except as hereinafter described.","FIG.","8 shows a hanger bracket 90 with arcuate slots 98 in complementary alignment located at a lower portion of first and second sides 92, 94 of the hanger bracket 90, elongate in a direction longitudinal of the trailing beam 20, and downwardly concave.","The first and second sides 92, 94 also contain pivot apertures 100 in complementary alignment, which are located vertically above the arcuate slots 98.An intermediate wall 96 is shown comprising a mounting aperture 102 at an upper portion thereof for mounting a linear actuator, such as a hydraulic or pneumatic ram (not shown).","Alternatively, the intermediate wall 96 can comprise an adjustment bolt aperture and a slot similar to the previously described adjustment bolt aperture 52 and slot 54 to accommodate an adjustment assembly comprising the previously described adjustment bolt 28.FIG.","9 shows an alternate pivot arm 104 complementary to the hanger bracket 90 of FIG.","8.The pivot arm 104 is a generally plate-like elongated member with a pivot portion 110 that tapers to a yoke end 106 and transitions to a trailing beam end 108.The pivot portion 110 contains a pivot stud 114 extending orthogonally therefrom and centered with respect to the longitudinal axis of the pivot arm 104.The yoke end 106 contains a shaft receptacle 116 centered with respect to the longitudinal axis of the pivot arm 104.The trailing beam end 108 has a pivot bolt aperture 112 centered with respect to the longitudinal axis of the pivot arm 104.The shaft receptacle 116 slidably receives the yoke stud 70.The pivot bolt aperture 112 slidably receives the pivot bolt 26.The pivot stud 114 is slidably received within the pivot aperture 100.The length of the pivot stud 114 is slightly greater than the thickness of a hanger bracket side 92, 94.The adjustment of the trailing arm alignment for the alternate embodiment is similar to the adjustment of the previously described preferred embodiment.","The adjustment bolt 28 is rotated, which draws the yoke assembly 39 fore and aft, depending upon the direction of rotation of the adjustment bolt 28.Movement of the yoke assembly 39 pivots the pivot arms 104 about the pivot stud 114.This moves the pivot bolt 26 along the arcuate slots 98 in an opposed fore-and-aft direction, which correspondingly moves the trailing beam 20 in a fore and aft direction.","This process is continued until the axle 16 and wheels 14 are properly aligned with the vehicle frame 12.When the axle 16 and wheels 14 are in the desired alignment, the nut 44 is tightened to secure the bushed connection 24 to the hanger bracket 22.It will be readily apparent to one of ordinary skill in the art that, if the distance between the pivot stud 114 and the shaft receptacle 116 is less than the distance between the pivot stud 114 and the pivot bolt aperture 112, the translational movement of the yoke assembly 39 will impart a proportionately greater translational movement to the pivot bolt 26, thereby providing a greater degree of adjustability than with the previously described preferred embodiment.","While the configuration and operation of the invention has been described with respect to embodiments comprising an adjustment bolt 28 and complementary yoke assembly 39, other embodiments capable of linear motion for translating a yoke assembly and providing fore-and-aft pivotable adjustment consistent with the inventive concepts described herein, such as a linear hydraulic, linear pneumatic, or electrically-powered actuator, can be utilized.","While particular embodiments of the invention have been shown, it will be understood, of course, that the invention is not limited thereto since modifications may be made by those skilled in the art, particularly in light of the foregoing teachings.","Reasonable variation and modification are possible within the scope of the foregoing disclosure of the invention without departing from the spirit of the invention."]],"string":"[\n [\n \"Trailing beam suspension with alignment adjustment\",\n \"A trailing beam suspension includes a mechanical adjustment mechanism for aligning the axle and wheels of a vehicle.\",\n \"The trailing beam is mounted to the vehicle frame by a pivot bolt passing through a hanger bracket and a conventional bushed connection.\",\n \"Opposing sides of the hanger bracket contain aligned curved apertures that receive the pivot bolt.\",\n \"The pivot bolt passes through pivot arms which are pivotally connected to the sides of the hanger bracket.\",\n \"The pivot arms are connected to a yoke to which an adjustment shaft is attached.\",\n \"Movement of the adjustment shaft moves the yoke fore and aft.\",\n \"The movement of the yoke pivots the pivot arms, thus moving the pivot bolt in a fore-and aft-direction.\",\n \"When the trailing beam has been aligned, the pivot connection is tightened, thus securing the trailing beam, axle, and wheels in the desired alignment.\"\n ],\n [\n \"1.In a trailing beam suspension for a vehicle comprising a hanger bracket adapted to be mounted to a vehicle frame rail, a trailing beam pivotally mounted at one end to the hanger bracket through a pivot pin and an adjustment slot in the hanger bracket so that the position of the beam with respect to the hanger bracket is selectively adjustable in a fore-and-aft direction to align an axle mounted to the trailing beam, the improvement comprising: a manually adjustable, threadably actuated drive mechanism mounted between the hanger bracket and the pivot bolt for incrementally adjusting the position of the pivot bolt in the slot to thereby adjust the beam with respect to the hanger bracket, wherein the drive mechanism includes a drive shaft that is substantially orthogonal to an axis of the pivot bolt and is substantially offset therefrom.\",\n \"2.The trailing beam suspension according to claim 1 wherein the pivot pin is mounted to at least one pivot arm, the pivot arm is pivotally mounted at one end to the hanger bracket for pivotal movement about an axis spaced from the adjustment slot and the drive mechanism is mounted to the at least one pivot arm at a location spaced from the adjustment slot and from the axis of the pivot arm, whereby pivotal movement of the at least one pivot arm selectively adjusts the relative position of the pivot bolt in the adjustment slot.\",\n \"3.The trailing beam suspension according to claim 2 wherein the drive mechanism further comprises a drive shaft connected to the at least one pivot arm to adjustably move the pivot arm as the shaft is driven.\",\n \"4.The trailing beam suspension according to claim 3 wherein the drive shaft is journaled in the hanger bracket and is threadably mounted to the at least one pivot arm.\",\n \"5.In a trailing beam suspension for a vehicle comprising a hanger bracket adapted to be mounted to a vehicle frame rail, a trailing beam pivotally mounted at one end to the hanger bracket through a pivot pin and an adjustment slot in the hanger bracket so that the position of the beam with respect to the hanger bracket is selectively adjustable in a fore-and-aft direction to align an axle mounted to the trailing beam, the improvement comprising: a drive mechanism mounted between the hanger bracket and the pivot bolt for incrementally adjusting the position of the pivot bolt in the slot to thereby adjust the beam with respect to the hanger bracket, wherein the pivot pin is mounted to at least one pivot arm, the pivot arm is pivotally mounted at one end to the hanger bracket for pivotal movement about an axis spaced from the adjustment slot and the drive mechanism is mounted to the at least one pivot arm at a location spaced from the adjustment slot and from the axis of the pivot arm, whereby pivotal movement of the at least one pivot arm selectively adjusts the relative position of the pivot bolt in the adjustment slot, the drive mechanism further comprising a drive shaft connected to the at least one pivot arm to adjustable move the pivot arm as the shaft is driven, and wherein the drive shaft being journaled in the hanger bracket and is threadably mounted to the at least one pivot arm and mounted to the at least one pivot arm through a yoke that has a threaded aperture in which the drive shaft is received.\",\n \"6.The trailing beam suspension according to claim 5 wherein the longitudinal axis of the shaft is orthogonal to the axis of the at least one pivot arm.\",\n \"7.The trailing beam suspension according to claim 6 wherein the hanger bracket has a pair of spaced side walls and an end wall joining the side walls, there are two pivot arms, one adjacent to each side wall of the hanger bracket and the drive shaft is journaled in the end wall.\",\n \"8.The trailing beam suspension according to claim 1 wherein the drive mechanism includes a threaded shaft.\",\n \"9.The trailing beam suspension according to claim 8 wherein rotation of the threaded shaft moves the pivot bolt in the slot.\",\n \"10.The trailing beam suspension according to claim 1 wherein the drive mechanism includes a linear hydraulic actuator.\",\n \"11.The trailing beam suspension according to claim 1 wherein the drive mechanism includes a linear pneumatic actuator.\",\n \"12.The trailing beam suspension according to claim 1 wherein the drive mechanism includes an electrically-powered actuator.\",\n \"13.The trailing beam suspension according to claim 7 wherein the shaft is substantially centrally located between the side walls of the hanger bracket.\",\n \"14.The trailing beam suspension according to claim 1 wherein the drive mechanism symmetrically engages the pivot bolt.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>1.Technical Field The invention relates to vehicle suspensions.\",\n \"In one aspect, the invention relates to a trailing beam suspension for a wheeled vehicle in which the suspension includes a mechanism for fore or aft adjustment of the alignment of the axle and wheels with respect to the longitudinal axis of the vehicle.\",\n \"2.Related Art Proper wheel alignment in road vehicles is important to prevent undue tire wear.\",\n \"Common causes of improper wheel alignment include improper initial suspension system installation, accident damage, and impact or shock loads imposed on suspension components by potholes or other irregularities in the road.\",\n \"Wheel alignment adjustment mechanisms permit adjustment of the alignment of vehicle axles and wheels with respect to the vehicle frame.\",\n \"Adjustment mechanisms are generally one of two types: (1) those that are adjustable as the suspension is installed on the vehicle and then permanently fixed in the adjusted position by welding or the like, and (2) those that are adjustable any time during and after installation and are held in the adjusted position mechanically.\",\n \"U.S. Pat.\",\n \"No.\",\n \"3,960,388 to Strader et al., which is incorporated herein in its entirety by reference, is an example of the first category of alignment devices.\",\n \"The Strader '388 patent discloses a trailing beam suspension that incorporates a mounting plate for the trailing beam having a pair of bosses that engage slots in the hanger bracket.\",\n \"The trailing beam is attached to the hanger bracket by a bolted connection in which the bolt passes through the mounting plate and is secured with a nut.\",\n \"The bolt head engages the mounting plate so that rotating the bolt pivots the mounting plate about one of the bosses.\",\n \"Pivoting of the mounting plate moves the bolt fore or aft, which moves the trailing beam fore or aft to align the axle.\",\n \"When the axle and wheels have been properly aligned, the plate is welded to the hanger bracket.\",\n \"Further adjustment is possible only after removing the weld.\",\n \"U.S. Pat.\",\n \"No.\",\n \"Re.\",\n \"30,308 to Masser discloses a trailing beam suspension system incorporating a mounting plate for the trailing beam having a single boss that engages a slot in the hanger bracket.\",\n \"The pivot bolt for the trailing beam passes through the mounting plate and slot.\",\n \"The trailing beam is moved fore or aft until proper alignment of the axle and wheels is achieved.\",\n \"The mounting plate is then welded to the hanger bracket.\",\n \"Further adjustment is possible only after removing the weld.\",\n \"U.S. Pat.\",\n \"No.\",\n \"4,991,872 to Richardson discloses a trailing beam suspension for a low-bed trailer.\",\n \"The forward trailing beam of a pair of trailing beams is mounted to the intermediate, inclined portion of the trailer's main structural beam through a bushing assembly.\",\n \"The bushing assembly is held to the web of the beam by a bolted connection.\",\n \"The web is provided with slotted mounting holes that enable adjustment of the alignment of the trailing beam and attached axle.\",\n \"The trailing beam is moved fore or aft until the axle and wheels are properly aligned.\",\n \"After adjustment, the bushing assembly is welded to the beam.\",\n \"Further adjustment requires removal of the weld and loosening of four bolts connecting the bushing assembly to the trailer beam.\",\n \"An example of the second category of alignment devices is disclosed in U.S. Pat.\",\n \"No.\",\n \"5,201,898 to Pierce.\",\n \"The Pierce '898 patent discloses a trailing beam suspension incorporating a pair of complementary discs that engage each other with a channel and rib arrangement, with the trailing beam mounting bolt passing therethrough.\",\n \"One disc is welded to the hanger bracket and incorporates an asymmetrical hole.\",\n \"The second disc contains an off-center hole and is rotatable relative to the first.\",\n \"The rotation of the second disc relative to the first is translated into lateral movement of the trailing beam mounting bolt.\",\n \"It is necessary to restrain the second disc against further rotation while the connection is tightened to prevent movement of the trailing beam from its aligned position.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>According to the invention, a trailing beam suspension for a vehicle comprises a hanger bracket adapted to be mounted to a vehicle frame rail, a trailing beam pivotally mounted at one end to the hanger bracket through a pivot pin and an adjustment slot in the hanger bracket so that the position of the beam with respect to the hanger bracket is selectively adjustable in a fore-and-aft direction to align an axle mounted to the trailing beam.\",\n \"According to the invention, a drive mechanism is mounted between the hanger bracket and the pivot bolt for incrementally adjusting the position of the pivot bolt in the slot to thereby adjust the beam with respect to the hanger bracket.\",\n \"In one embodiment, the pivot pin is mounted to at least one pivot arm, the pivot arm is pivotally mounted at one end to the hanger bracket for pivotal movement about an axis spaced from the adjustment slot and the drive mechanism is mounted to the at least one pivot arm at a location spaced from the adjustment slot and from the axis of the pivot arm.\",\n \"Pivotal movement of the at least one pivot arm selectively adjusts the relative position of the pivot bolt in the adjustment slot.\",\n \"Preferably, the drive mechanism further comprises a drive shaft connected to the at least one pivot arm to adjustably move the pivot arm as the shaft is driven.\",\n \"In one embodiment, the drive shaft is journaled in the hanger bracket and is threadably mounted to the at least one pivot arm.\",\n \"Further, in a preferred embodiment, the drive shaft is mounted to the at least one pivot arm through a yoke that has a threaded aperture in which the drive shaft is received.\",\n \"Typically, the longitudinal axis of the shaft is orthogonal to the axis of the at least one pivot arm.\",\n \"In a preferred embodiment, the hanger bracket has a pair of spaced side walls and an end wall joining the side walls.\",\n \"There are two pivot arms, one adjacent to each side wall of the hanger bracket and the drive shaft is journaled in the end wall of the hanger bracket.\",\n \"In a preferred embodiment of the invention, the drive mechanism includes a threaded shaft and rotation of the threaded shaft moves the pivot bolt in the slot.\",\n \"In another embodiment, the drive mechanism includes a linear hydraulic actuator.\",\n \"In still another embodiment, the drive mechanism includes a linear pneumatic actuator.\",\n \"In yet another alternative embodiment, the drive mechanism includes an electrically-powered actuator.\"\n ],\n [\n \"CROSS-REFERENCE TO RELATED APPLICATION This application claims the benefit of U.S.\",\n \"Provisional Patent Application Ser.\",\n \"No.\",\n \"60/288,935, filed on May 4, 2001.BACKGROUND OF THE INVENTION 1.Technical Field The invention relates to vehicle suspensions.\",\n \"In one aspect, the invention relates to a trailing beam suspension for a wheeled vehicle in which the suspension includes a mechanism for fore or aft adjustment of the alignment of the axle and wheels with respect to the longitudinal axis of the vehicle.\",\n \"2.Related Art Proper wheel alignment in road vehicles is important to prevent undue tire wear.\",\n \"Common causes of improper wheel alignment include improper initial suspension system installation, accident damage, and impact or shock loads imposed on suspension components by potholes or other irregularities in the road.\",\n \"Wheel alignment adjustment mechanisms permit adjustment of the alignment of vehicle axles and wheels with respect to the vehicle frame.\",\n \"Adjustment mechanisms are generally one of two types: (1) those that are adjustable as the suspension is installed on the vehicle and then permanently fixed in the adjusted position by welding or the like, and (2) those that are adjustable any time during and after installation and are held in the adjusted position mechanically.\",\n \"U.S. Pat.\",\n \"No.\",\n \"3,960,388 to Strader et al., which is incorporated herein in its entirety by reference, is an example of the first category of alignment devices.\",\n \"The Strader '388 patent discloses a trailing beam suspension that incorporates a mounting plate for the trailing beam having a pair of bosses that engage slots in the hanger bracket.\",\n \"The trailing beam is attached to the hanger bracket by a bolted connection in which the bolt passes through the mounting plate and is secured with a nut.\",\n \"The bolt head engages the mounting plate so that rotating the bolt pivots the mounting plate about one of the bosses.\",\n \"Pivoting of the mounting plate moves the bolt fore or aft, which moves the trailing beam fore or aft to align the axle.\",\n \"When the axle and wheels have been properly aligned, the plate is welded to the hanger bracket.\",\n \"Further adjustment is possible only after removing the weld.\",\n \"U.S. Pat.\",\n \"No.\",\n \"Re.\",\n \"30,308 to Masser discloses a trailing beam suspension system incorporating a mounting plate for the trailing beam having a single boss that engages a slot in the hanger bracket.\",\n \"The pivot bolt for the trailing beam passes through the mounting plate and slot.\",\n \"The trailing beam is moved fore or aft until proper alignment of the axle and wheels is achieved.\",\n \"The mounting plate is then welded to the hanger bracket.\",\n \"Further adjustment is possible only after removing the weld.\",\n \"U.S. Pat.\",\n \"No.\",\n \"4,991,872 to Richardson discloses a trailing beam suspension for a low-bed trailer.\",\n \"The forward trailing beam of a pair of trailing beams is mounted to the intermediate, inclined portion of the trailer's main structural beam through a bushing assembly.\",\n \"The bushing assembly is held to the web of the beam by a bolted connection.\",\n \"The web is provided with slotted mounting holes that enable adjustment of the alignment of the trailing beam and attached axle.\",\n \"The trailing beam is moved fore or aft until the axle and wheels are properly aligned.\",\n \"After adjustment, the bushing assembly is welded to the beam.\",\n \"Further adjustment requires removal of the weld and loosening of four bolts connecting the bushing assembly to the trailer beam.\",\n \"An example of the second category of alignment devices is disclosed in U.S. Pat.\",\n \"No.\",\n \"5,201,898 to Pierce.\",\n \"The Pierce '898 patent discloses a trailing beam suspension incorporating a pair of complementary discs that engage each other with a channel and rib arrangement, with the trailing beam mounting bolt passing therethrough.\",\n \"One disc is welded to the hanger bracket and incorporates an asymmetrical hole.\",\n \"The second disc contains an off-center hole and is rotatable relative to the first.\",\n \"The rotation of the second disc relative to the first is translated into lateral movement of the trailing beam mounting bolt.\",\n \"It is necessary to restrain the second disc against further rotation while the connection is tightened to prevent movement of the trailing beam from its aligned position.\",\n \"SUMMARY OF THE INVENTION According to the invention, a trailing beam suspension for a vehicle comprises a hanger bracket adapted to be mounted to a vehicle frame rail, a trailing beam pivotally mounted at one end to the hanger bracket through a pivot pin and an adjustment slot in the hanger bracket so that the position of the beam with respect to the hanger bracket is selectively adjustable in a fore-and-aft direction to align an axle mounted to the trailing beam.\",\n \"According to the invention, a drive mechanism is mounted between the hanger bracket and the pivot bolt for incrementally adjusting the position of the pivot bolt in the slot to thereby adjust the beam with respect to the hanger bracket.\",\n \"In one embodiment, the pivot pin is mounted to at least one pivot arm, the pivot arm is pivotally mounted at one end to the hanger bracket for pivotal movement about an axis spaced from the adjustment slot and the drive mechanism is mounted to the at least one pivot arm at a location spaced from the adjustment slot and from the axis of the pivot arm.\",\n \"Pivotal movement of the at least one pivot arm selectively adjusts the relative position of the pivot bolt in the adjustment slot.\",\n \"Preferably, the drive mechanism further comprises a drive shaft connected to the at least one pivot arm to adjustably move the pivot arm as the shaft is driven.\",\n \"In one embodiment, the drive shaft is journaled in the hanger bracket and is threadably mounted to the at least one pivot arm.\",\n \"Further, in a preferred embodiment, the drive shaft is mounted to the at least one pivot arm through a yoke that has a threaded aperture in which the drive shaft is received.\",\n \"Typically, the longitudinal axis of the shaft is orthogonal to the axis of the at least one pivot arm.\",\n \"In a preferred embodiment, the hanger bracket has a pair of spaced side walls and an end wall joining the side walls.\",\n \"There are two pivot arms, one adjacent to each side wall of the hanger bracket and the drive shaft is journaled in the end wall of the hanger bracket.\",\n \"In a preferred embodiment of the invention, the drive mechanism includes a threaded shaft and rotation of the threaded shaft moves the pivot bolt in the slot.\",\n \"In another embodiment, the drive mechanism includes a linear hydraulic actuator.\",\n \"In still another embodiment, the drive mechanism includes a linear pneumatic actuator.\",\n \"In yet another alternative embodiment, the drive mechanism includes an electrically-powered actuator.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS The invention will be described with reference to the accompanying drawings in which: FIG.\",\n \"1 is a side elevational view of a trailing beam suspension incorporating the present invention.\",\n \"FIG.\",\n \"2 is a top perspective view of a preferred embodiment of the trailing beam suspension of FIG.\",\n \"1 showing a yoke assembly installed in a hanger bracket and a trailing beam pivot connection.\",\n \"FIG.\",\n \"3 is a perspective view of the hanger bracket illustrated in FIG.\",\n \"2.FIG.\",\n \"4 is a perspective view of a portion of the yoke assembly illustrated in FIG.\",\n \"2.FIG.\",\n \"5 is a perspective view of a pivot arm illustrated in FIG.\",\n \"2.FIG.\",\n \"6 is a side perspective view of a subassembly consisting of the yoke assembly, pivot arms, and pivot bolt illustrated in FIG.\",\n \"2.FIG.\",\n \"7 is a side view of the subassembly of FIG.\",\n \"6 installed in the hanger bracket and illustrating the relative movement of the components of the alignment mechanism as the device is operated.\",\n \"FIG.\",\n \"8 is a perspective view of a hanger bracket comprising part of an alternate embodiment of the trailing beam suspension of FIG.\",\n \"1.FIG.\",\n \"9 is a perspective view of a pivot arm which is complementary to the hanger bracket illustrated in FIG.\",\n \"8.DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Referring now to the drawings, and in particular to FIG.\",\n \"1, there is illustrated a trailing beam suspension system for a vehicle generally identified by the numeral 10.The suspension system comprises a trailing beam 20, a hanger bracket 22, and an air spring 30.The hanger bracket 22 is attached in a conventional manner to a vehicle frame rail 12.A first end of the trailing beam 20 is pivotally mounted to the hanger bracket 22 through a pivot bolt 26 and a conventional bushed connection (not shown) in the same manner as the trailing arm 22 is pivotally mounted to the bolt 34 in the Strader et al.\",\n \"U.S. Pat.\",\n \"No.\",\n \"3,960,388.A second end of the trailing beam 20 is attached in a conventional manner to the air spring 30 so that the air spring 30 is disposed in load transmitting relation between the trailing beam 20 and the frame rail 12.An axle 16, with attached wheels 14, is connected to the trailing beam 20 by an axle bracket 18.Referring now to FIGS.\",\n \"2-6, a preferred embodiment of an alignment mechanism generally comprises the hanger bracket 22, a pivot pin 25 comprising a threaded pivot bolt 26 with a complementary nut 44, an adjustable yoke assembly 39, and two pivot arms 46 with attached pivot studs 38.The hanger bracket has two parallel, laterally spaced-apart sides comprising a first side 56 and a second side 58 joined by an intermediate side 60 generally orthogonal thereto.\",\n \"Washers 42 are spaced between the outer faces of the hanger bracket sides 56, 58, and the head of the pivot bolt 26 and nut 44, respectively.\",\n \"Referring specifically to FIG.\",\n \"3, the first and second sides 56, 58 of the hanger bracket 22 contain arcuate slots 62 in complementary alignment located at a lower portion of the hanger bracket 22.The arcuate slots 62 are elongate in a direction longitudinal of the trailing beam 20, and upwardly concave.\",\n \"The first and second sides 56, 58 of the hanger bracket 22 also contain pivot apertures 64 in complementary alignment, which are located vertically beneath the arcuate slots 62 proximate to the lower edge of the hanger bracket 22.The arcuate slots 62 slidably receive the pivot bolt 26 and to allow the pivot bolt 26 to laterally translate therein.\",\n \"The pivot apertures 64 slidably receive the pivot studs 38 and to allow rotation of the pivot studs 38 therein.\",\n \"In the embodiment described herein, the upper portion of the intermediate side 60 contains an adjustment bolt aperture 52 and a slot 54 extending generally downwardly from the adjustment bolt aperture 52.The slot 54 accommodates the shaft of an adjustment bolt 28 to enable the adjustment bolt 28 to be freely turned therein.\",\n \"FIGS.\",\n \"2, 4 and 6 show the yoke assembly 39 of the preferred embodiment comprising the adjustment bolt 28 with a bolt head 29, a retainer nut 50, a yoke 40 having a threaded yoke aperture 72, and a yoke nut 48.As hereinafter described, turning of the adjustment bolt 28 in the yoke aperture 72 translates the yoke 40 to pivot the pivot arms 46 about the pivot studs 38.Alternate assemblies for urging the pivot arms 46 to pivot about the pivot studs 38 can be employed, such as a linear hydraulic, linear pneumatic, or electrically-powered actuator.\",\n \"For example, a linear hydraulic actuator can have a hydraulic piston with a piston rod attached to the pivot arms 46 through an articulating connection so that extension and retraction of the hydraulic piston and piston rod pivots the pivot arms 46 fore and aft.\",\n \"In the embodiment shown in FIGS.\",\n \"2, 4 and 6, the retainer nut 50 is threaded onto the adjustment bolt 28 until it is proximate to the bolt head 29, leaving a gap between the retainer nut 50 and bolt head 29 that is slightly larger than the thickness of the intermediate side 60.The retainer nut 50 is fixedly attached to the adjustment bolt 28, such as by welding or brazing.\",\n \"The yoke 40 comprises a hub 66, opposing arms 68, and a yoke aperture 72.The hub 66 is generally cylindrical and contains the yoke aperture 72 extending axially therethrough.\",\n \"Extending radially from the hub 66 are the diametrically opposed arms 68.The arms 68 are generally cylindrical, axially aligned, and terminate in a pair of spaced-apart axially-opposed yoke studs 70.The yoke stud 70 is a cylindrical projection with a diameter that is smaller than the arm diameter and which is axially aligned with the arm 68.The length of the yoke stud 70 is slightly less than the thickness of the pivot arm 46.A recess 74 extends across the center portion of the hub 66 intermediate the arms 68.The recess 74 accommodates the yoke nut 48 to restrain the yoke nut 48 against rotation relative to the yoke 40.The yoke nut 48 is axially aligned with the yoke aperture 72 and is fixedly attached to the yoke 40, such as by welding or brazing.\",\n \"The yoke 40 and yoke nut 48 can be threaded onto the adjustment bolt 28 by first passing the adjustment bolt 28 through the yoke aperture 72 and into the yoke nut 48.Referring now to FIG.\",\n \"5, the pivot arm 46 is a generally plate-like elongated member with a central portion 84 that tapers to a first end 76 and to a second end 78.The central portion 84 contains a pivot bolt aperture 82 centered with respect to the longitudinal axis of the pivot arm 46.The first end 76 contains a shaft receptacle 80 centered with respect to the longitudinal axis of the pivot arm 46.The second end 78 has a pivot stud 38 extending orthogonally therefrom and centered with respect to the longitudinal axis of the pivot arm 46.The shaft receptacle 80 slidably receives the yoke stud 70.The pivot bolt aperture 82 slidably receives the pivot bolt 26.The pivot stud 38 is slidably received within the pivot aperture 64.The length of the pivot stud 38 is slightly greater than the thickness of a hanger bracket side 56, 58.Referring now to FIGS.\",\n \"6 and 7, the assembly and operation of the invention will be described.\",\n \"The adjustment bolt 28 is inserted through the yoke aperture and threaded into the yoke nut 48.The yoke studs 70 are inserted into the shaft receptacles 80 through the pivot arms 46, enabling each pivot arm 46 to pivot about its respective yoke stud 70.The pivot studs 38 are inserted into the pivot apertures 64 in the hanger bracket sides 56, 58, enabling the pivot arms 46 to pivot about the pivot studs 38.The adjustment bolt head 29 is passed through the adjustment bolt aperture 22, and the adjustment bolt 28 is positioned in the slot 54 so that the bolt head 29 is to the outside of the intermediate side 60 and the retainer nut 50 is to the inside of the intermediate side 60.The adjustment bolt 28 can be turned freely in the slot 54 and is restrained against axial movement by the bolt head 29 and retainer nut 50.Furthermore, the adjustment bolt 28 can move vertically within the slot 54 during the adjustment process.\",\n \"The pivot bolt apertures 82 are also aligned with the arcuate slots 62.The pivot bolt 26 is inserted through a washer 42, a pivot bolt aperture 82, an arcuate slot 62, the bushed connection 24 of the trailing beam, the second arcuate slot 62, the second pivot bolt aperture 82, the second washer 42, and threaded into the nut 44 to pivotally connect the trailing beam to the hanger bracket 22.As assembled, each pivot arm 46 is in slidable contact with the inside face of its respective hanger bracket side 56, 58.After the alignment subassembly has been assembled in the hanger bracket 22 with the trailing beam 20 mounted to the pivot bolt 26 as described above, the alignment of the trailing beam is accomplished as follows.\",\n \"The pivot bolt 26 and nut 44 are in a slightly loosened configuration.\",\n \"The adjustment bolt 28 is rotated, which draws the yoke assembly 39 fore or aft, depending upon the direction of rotation of the adjustment bolt 28.Movement of the yoke assembly 39 pivots the pivot arms 46 about the pivot stud 38.This moves the pivot bolt 26 along the arcuate slots 62 in a corresponding fore or aft direction, which correspondingly moves the trailing beam 20 in a fore or aft direction.\",\n \"This process is continued until the axle 16 and wheels 14 are properly aligned with the vehicle frame 12.When the axle 16 and wheels 14 are in the desired alignment, the nut 44 is tightened to secure the bushed connection 24 to the hanger bracket 22.The trailing beam 20, the axle 16, and the wheels 14 can be easily and accurately aligned due to the unique mechanism.\",\n \"Additionally, the pivot bolt nut 44 can be securely tightened to the proper torque without the risk that the trailing beam 20 will become misaligned during the tightening process.\",\n \"Tightening the pivot bolt nut 44 to the proper torque reduces the risk that the nut 44 will loosen over time and the pivot connection 24 will fail due to under-torquing of the nut 44.Additionally, the pivot bolt 26 is securely restrained against lateral movement in the slots 62 unless the adjustment bolt 28 is turned.\",\n \"The unique alignment mechanism thus provides further protection against misalignment of the trailing beam 20, axle 16, and wheels 14 due to impact or shock loads imposed on the suspension by potholes or other road surface irregularities.\",\n \"The trailing beam 20, axle 16, and wheels 14 will remain in the properly aligned position even if the nut 44 becomes loosened.\",\n \"Furthermore, the alignment of the trailing beam 20 can be maintained without the necessity of welding the alignment components to the hanger bracket or vehicle frame, thus facilitating future adjustment to the wheel alignment.\",\n \"An alternate embodiment of the invention comprises a hanger bracket with alternate arcuate slots and pivot apertures which is generally similar to the hanger bracket 22, except as hereinafter described.\",\n \"FIG.\",\n \"8 shows a hanger bracket 90 with arcuate slots 98 in complementary alignment located at a lower portion of first and second sides 92, 94 of the hanger bracket 90, elongate in a direction longitudinal of the trailing beam 20, and downwardly concave.\",\n \"The first and second sides 92, 94 also contain pivot apertures 100 in complementary alignment, which are located vertically above the arcuate slots 98.An intermediate wall 96 is shown comprising a mounting aperture 102 at an upper portion thereof for mounting a linear actuator, such as a hydraulic or pneumatic ram (not shown).\",\n \"Alternatively, the intermediate wall 96 can comprise an adjustment bolt aperture and a slot similar to the previously described adjustment bolt aperture 52 and slot 54 to accommodate an adjustment assembly comprising the previously described adjustment bolt 28.FIG.\",\n \"9 shows an alternate pivot arm 104 complementary to the hanger bracket 90 of FIG.\",\n \"8.The pivot arm 104 is a generally plate-like elongated member with a pivot portion 110 that tapers to a yoke end 106 and transitions to a trailing beam end 108.The pivot portion 110 contains a pivot stud 114 extending orthogonally therefrom and centered with respect to the longitudinal axis of the pivot arm 104.The yoke end 106 contains a shaft receptacle 116 centered with respect to the longitudinal axis of the pivot arm 104.The trailing beam end 108 has a pivot bolt aperture 112 centered with respect to the longitudinal axis of the pivot arm 104.The shaft receptacle 116 slidably receives the yoke stud 70.The pivot bolt aperture 112 slidably receives the pivot bolt 26.The pivot stud 114 is slidably received within the pivot aperture 100.The length of the pivot stud 114 is slightly greater than the thickness of a hanger bracket side 92, 94.The adjustment of the trailing arm alignment for the alternate embodiment is similar to the adjustment of the previously described preferred embodiment.\",\n \"The adjustment bolt 28 is rotated, which draws the yoke assembly 39 fore and aft, depending upon the direction of rotation of the adjustment bolt 28.Movement of the yoke assembly 39 pivots the pivot arms 104 about the pivot stud 114.This moves the pivot bolt 26 along the arcuate slots 98 in an opposed fore-and-aft direction, which correspondingly moves the trailing beam 20 in a fore and aft direction.\",\n \"This process is continued until the axle 16 and wheels 14 are properly aligned with the vehicle frame 12.When the axle 16 and wheels 14 are in the desired alignment, the nut 44 is tightened to secure the bushed connection 24 to the hanger bracket 22.It will be readily apparent to one of ordinary skill in the art that, if the distance between the pivot stud 114 and the shaft receptacle 116 is less than the distance between the pivot stud 114 and the pivot bolt aperture 112, the translational movement of the yoke assembly 39 will impart a proportionately greater translational movement to the pivot bolt 26, thereby providing a greater degree of adjustability than with the previously described preferred embodiment.\",\n \"While the configuration and operation of the invention has been described with respect to embodiments comprising an adjustment bolt 28 and complementary yoke assembly 39, other embodiments capable of linear motion for translating a yoke assembly and providing fore-and-aft pivotable adjustment consistent with the inventive concepts described herein, such as a linear hydraulic, linear pneumatic, or electrically-powered actuator, can be utilized.\",\n \"While particular embodiments of the invention have been shown, it will be understood, of course, that the invention is not limited thereto since modifications may be made by those skilled in the art, particularly in light of the foregoing teachings.\",\n \"Reasonable variation and modification are possible within the scope of the foregoing disclosure of the invention without departing from the spirit of the invention.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476914"}}},{"rowIdx":982,"cells":{"text":{"kind":"list like","value":[["FILTER TOW","Abstract of the Disclosure The invention relates to a filter tow made from crimped endless cellulose acetate filaments, whereby the filaments form at least two connected fibre bundles with the same filament titre of at least 600 filaments.","The filament titres differ from bundle to bundle by at least 1 dtex Furthermore a filter tow is disclosed with a crimp ratio of more than 40%, said filaments comprising at least 22 crimp turns per filament length."],["1.A filter tow of crimped, continuous cellulose acetate filaments, wherein the filaments form at least two coherent fiber bundles of the same filament dernier of at least 600 filaments which differ from fiber bundle to fiber bundle in their filament dernier by at least 1 dtex.","2.The filter tow of claim 1, wherein the filaments form more than five coherent fiber bundles of the same filament dernier.","3.The filter of claim 1 or 2, wherein the individual fiber bundles contain more than 1000 filaments.","4.The filter tow of one of claims 1 to 3, wherein the filament derniers of the filaments differ from fiber bundle to fiber bundle by more than 1.5 dtex.","5.The filter tow of one of claims 1 to 4, wherein the crimp ratio is greater than 33%, and the number of crimps per cm is greater than 15.6.The filter tow of one of claims 1 to 5, wherein the crimp ratio is greater than 35% 7.The filter tow of one of the above claims, wherein the crimp ratio is greater than 40%, preferably greater than 42%, and the number of crimps per cm exceeds a value of 22.8.The filter tow of crimped, continuous cellulose acetate filaments, wherein the filter tow has a crimp ratio of greater than 40% and the filaments have at least 22 crimps per cm filament length.","9.The filter tow of claim 8, wherein the crimp ratio is greater than 42% and the filaments have more than 25 crimps per cm.","10.The filter tow of claim 8 or 9, wherein the filament dernier is greater than 3.3 dtex, and the total dernier is less than 44,000 dtex, preferably less than 40,000 dtex."],["Detailed Description of the Invention DESCRIPTION The invention relates to a filter tow used in the manufacture of cigarette filters having a comparatively low filtration performance whose draw resistance retention characteristic was selected in such a way that their use in full-flavor cigarettes with a ventilation degree of more than 25% is possible while maintaining an acceptable draw resistance in the cigarette and an acceptable filter plug weight.","The great majority of cigarette filters currently in use are manufactured from filter tow, which consists of continuous cellulose 2,5-acetate filaments.","These filters are referred to as acetate filters.","The cigarettes that are manufactured and sold in the European Union can be classified as follows based on their condensate contents in smoke: Full-flavor Medium Light, and Ultralight cigarettes Because of the maximum-content regulations (90/239/EEC) applying to condensate content, all of these cigarettes are ventilated.","The degree of ventilation depends on the condensate target value and on the filtration performance of the filter being used.","The following table summarizes the typical current values: [0009.1] Class mg Condensate Filament Dernier % Ventilation Full Flavor <12 2.8-3.3 dtex <25 Medium 8-10 ≦3.3 dtex <35 Light 4-8 2.1-2.8 dtex 30-50 Ultralight <41 1.7-8.8 >50 Most of products currently sold have a diameter of 7.8.+-.0.2 mm and a cigarette length of 85-120 mm.","In addition, there are niche products whose design characteristics differ significantly from the most commonly sold products.","In response to this wide variety of product requirements, the filter tow industry supplies a multitude of different tow types that can essentially be described by the following characteristics: filament dernier (dtex) 1.6-8.8; total dernier (dtex) 17,000-60,000; crimp index (%) 25 max.","38; number of crimps per cm: 10 max.","20.The following description of the object of the invention exclusively relates to the class of full-flavor products.","A typical filter cigarette of the full-flavor class has the following typical characteristics (these values are approximate typical values).","They serve simply as a basis for calculations for further embodiments.","Lengths: Cigarette:84 mm Filter:24 mm Tobacco rod:63 mm Tipping:26 mm Diameters: Filter:7.80 mm Cigarette:7.85 mm Draw resistance: Filter:66 mm water column Tobacco rod:50 mm water column Cigarette:100 mm water column Filter tow specification: 3.0 Y 35 (filament dernier: 3.33 dtex; total dernier: 38.900, cross-sectional shape of the filaments: Y) Tobacco mixture: American Blend Ventilation: Degree of ventilation:20% Position of ventilation zone:11 mm from the mouth end Smoke values: Condensate:12 mg Nicotine:0.9 mg Carbon monoxide (CO): 14 mg Retention values: Condensate:42% Nicotine:38% Carbon monoxide (CO):None As a consequence of the new maximum-content regulation (2000/C 150E/03) of the European Union, full-flavor products in particular will have to undergo drastic changes in their design features, since effective Jan. 1, 2004 not only the condensate value, but also the nicotine value--and in particular the CO value--will be regulated with respect to their maximum limits.","The challenge involving the modification of the products to meet the new EU regulations will be that the permitted maximum amounts with an optimal smoke flavor will need to hit the values 10 mg condensate, 1 mg nicotine, and 10 mg CO per cigarette as precisely as possible.","In other words, in the future a 1:1 ratio of CO/tar must be targeted for full-flavor products.","As the above table shows, the ratio in this example is 1.17.Based on the model calculations that solely take a change in filter ventilation into account, it can be assumed that a filter ventilation of approximately 35% will be required to reduce the CO from the current value in the above calculation example of 14 mg to a future value of 10 mg per cigarette.","An increase in the degree of ventilation in an otherwise unchanged product will unavoidably necessitate a decrease in the cigarette draw resistance.","For consumers however, an optimal smoke flavor means that the draw resistance of the cigarette must not fall significantly below the current value of approximately 100 mm water column.","Increasing the degree of ventilation, however, will mean that maintaining the other filter parameters such as draw resistance, diameter, position of the ventilation zone, and the filter length will significantly reduce the draw resistance of the cigarette.","The resulting cigarette draw resistance can be calculated using the following equation, which is based on a laminar flow model: Pv=Po-Vx(Po-PfxLv/Lf), where: Pv: Draw resistance of the ventilated cigarette (mm water column) Po: Draw resistance of unventilated cigarette: =draw resistance [filter+tobacco rod] (mm water column) Pf: Filter draw resistance (mm water column) V: Ventilation degree of filter (%)/100 Lv: Distance of the ventilation zone from the mouth end (mm) Lf: Filter length Thus, the open draw resistance will then only be 88 mm water column (instead of the original 100 mm water column) for the stated sample cigarette at a ventilation degree of 35%.","Such a drop in the cigarette draw resistance is clearly perceivable for the smoker and it must be offset by an increase in the filter resistance (in the above example to approximately 80 mm water column) in order to maintain the consumer's smoke feel.","This increase in the filter draw resistance results in an unavoidable increase in the CO/tar ratio, since the tar values, but not the CO values, are influenced by the filter draw resistance (an increase in the filter draw resistance results in a decrease in the tar values).","Therefore, the draw resistance/retention ratios must be redefined for future products in the \"full-flavor\" category.","The adjustment to a 1:1 CO/tar ratio and the filter tow used to manufacture the filter needed to achieve an acceptable product with regard to cigarette draw resistance must be modified to meet these requirements.","In this regard, the following specifications may be defined: 1.The filter tow must be suitable for producing the draw resistance values that are typical today (however preferably even higher values).","2.The filtration performance of the filter made with the filter tow of the invention must, with the given filter draw resistance, be significantly lower than the current typical retention values in order to achieve the 1:1 CO/tar ratio.","3.The additional material consumption that results from the increase of the filament dernier must be kept within acceptable limits.","With a given filter diameter and length, the retention of a filter depends exclusively on the draw resistance and the filament dernier.","For example, an increase in the filament dernier and a decrease in the draw resistance results in a drop in filtration performance (and vice-versa).","Since the interrelationships are of a very complex nature, filter calculation programs are used to estimate the extent of such changes.","One example of such calculation programs is Cabled (Capability Line Expert; copyright Rhodia Acetow GmbH 1998, Freiburg, Germany) from the firm of Rhodia Acetow.","In the prior art, therefore, the retention of a filter is reduced by increasing the filament dernier.","Model calculations suggest that in the future the filament dernier will need to be increased by at least 0.5-1.5 dtex in order to meet the requirements for filtration performance.","However, the filter tow specifications commonly encountered in the market (the mean total dernier of all specifications is approximately 35 ktex) are not suitable for also covering the required draw resistance range with the required minimum filament dernier (retention) (Cable.COPYRGT.","calculation program from Rhodia Acetow GmbH).","In the prior art, this problem can only be solved with a monoacetate filter by drastically increasing the filter dernier to well above 45,000 dtex.","Such solutions, for example a filter tow of the type 4.5 Y 55 (see FIG.","2) in the prior art would then meet the draw resistance and retention specifications, but would result in an increase in the consumption of filter tow of >35% compared with current products.","The above discussion was limited to a description of the full-flavor group of cigarettes.","If, however, a CO/tar ratio of 1:1 is generally sought, the discussion regarding the widening of the filter resistance ranges would apply to all filament derniers.","This means that the object of the invention described below will generally comprise the currently standard filament dernier spectrum of 1.6-8.8 dtex.","Therefore, the object of the invention is to disclose a filter tow with which it is possible to manufacture filters that have a filtration performance achieving a CO/tar ratio of 1:1.In order to meet the above requirements, the invention proposes two approaches that are basically independent of each other and whose elements can, if desired, be combined.","The solution of the invention relates to a filter tow of crimped, continuous cellulose acetate filaments, in which the filaments form at least two coherent fiber bundles that have the same filament dernier and have at least 600 filaments and that differ in their filament dernier from fiber bundle to fiber bundle by at least 1 dtex.","In particular, this solution produces outstanding results with regard to filtration performance, since it was unexpectedly found that a filter that is manufactured from such a non-homogeneously distributed mixed tow exhibits a significantly lower tar retention than does a product made of a conventional tow with a uniform filament dernier distribution or a mixed tow having a homogeneous filament distribution (see FIG.","3).","The second solution is accomplished by a filter tow that has a more intensive and at the same time markedly finer crimp structure than the typical products encountered today.","Claim 4 recites a filter tow having a crimp index of more than 40%, in which the filaments have at least 22 curls per cm filament length.","In particular, this filter tow fulfills the maximum requirements with regard to filter draw resistance.","The values that are achieved are even exceeded if the crimp index is more than 42%, whereby the filaments preferably have more than 25 curls per cm.","In order at the same time to achieve optimal filtration performance by the filters, a filter tow having a filament dernier of more than 3.33 dtex and a total dernier of less than 44,000 dtex, preferably less than 40,000 dtex is proposed.","The limitation on the total dernier results in an increased filter tow consumption that is still acceptable.","The design and the advantages of the two solutions of the invention may be described as follows: The crimp index Ix is a measure of the intensity of curling.","The crimp index of a filter tow is determined by means of the tensile test (load/elongation relationship).","It is defined as the ratio of the elongated length L2 under the test load minus the initial length over the initial length under preload L1: Ix=(L2-L1)/L1*100(%) Here, the test load is 25 N and the preload is 2.5 N. The length between the jaws of the tensile test machine is 250 mm.","The curling index is determined in a tensile test at a constant traverse rate of 300 mm/min on a G02 test machine manufactured by Borgwaldt GmbH, Hamburg.","10 individual readings were taken per measurement.","Testing is performed in a standard climate: 20.degree.","C. and 60% relative humidity.","(The test method for determining the crimp ratio is described in the Cable program under the \"Help\" button).","The crimp ratio values of current filter tow specifications normally lie between 25 and 35%.","The firm of Rhodia Acetow is the sole manufacturer that offers worldwide specifications under the designation SK in a filament dernier range of <2.78 dtex that covers a crimp ratio range of from greater than 35% to a maximum of 38%.","As already stated, the crimp ratio is a measure of the intensity of crimping and, thus, the position of the performance curve.","In this context, a performance curve is understood to mean a performance range of a filter tow, which states the draw resistance range in which a filter tow can be processed with a given diameter and filter rod length under typical processing conditions.","For this working range, the performance curve describes the relationship between the draw resistance and weight.","The relationship between the position of the performance curve and the crimp ratio is shown using the example of a filter tow of type 2.1 Y 35 HK (filament dernier: 2.33 dtex; total demier: 38,900; cross-sectional shape of the filaments: Y; crimp ratio: 32%) and a 2.1 Y 35 SK (filament dernier: 2.33 dtex; total demier: 38,900, cross-sectional shape of the filament: Y; crimp ratio: 36%) in FIG.","1.The above object is accomplished by manufacturing a filter tow that has a crimp ratio of more than 40%, preferably more than 42%.","This ratio is absolutely necessary in order to cover the corresponding draw resistance range.","The crimp ratio is affected by the frequency and the amplitude of the crimps.","The number of crimps provides a measure of the fineness of crimping.","Currently, typical values range from 10 to max.","20 crimps per cm filament length.","The SK tows referred to above normally have 15-18 crimps per cm.","It was unexpectedly discovered that such an index can only be achieved with an acceptable filter tow tensile strength if the crimping is hyper-fine.","Therefore, the filter tow of the invention has, in addition to an index of more than 40% in the filaments, on average more than 22 crimps per cm, preferably more than 25.Such a filter tow can be processed on conventional filter rod machines into cigarette filters that have a draw resistance level comparable to filters manufactured using the currently typical specifications.","FIG.","2 shows the working range of a tow of type 4.5 Y 35 of the invention (filament dernier: 5.0 dtex; total demier: 38,900 dtex) in comparison with a conventional filter tow of type 3.0 Y 35 (filament dernier 3.33 dtex: total dernier: 38,900 dtex) respectively a conventional 4.5 Y 55 and a conventional 4.5 Y 35.FIG.","2 clearly shows that, compared with the current situation (3.0 Y 35) significantly more material is consumed with the solution of the invention (Example 1: 4.5 Y 35) as a result of the increase of the filament dernier, but that this additional consumption ends up being significantly less than with a traditional prior-art solution (4.5 Y 55).","In order to produce hyper-fine crimping without being overly aggressive, certain basic conditions must be met in the manufacturing process.","Prior to the crimping process, the filter tow band contains the following components that soften the acetate filaments: water, acetone, and softening components.","It is known that these softening components have a lasting effect on the crimping process.","For example, U.S. Pat.","No.","5,225,277 describes the effect of acetone and tow band temperatures on the stability of crimping.","Products having up to 19.7 crimps per cm can be manufactured using the method described therein.","This document does not make any statements with regard to the crimp ratio.","However, using the data stated therein (draw resistance, tow specification and machine settings) a crimp ratio that is clearly below 35% can be estimated with the help of the Cable software referred to above.","JP unexamined patent document 56-53223 describes the effects of steam-treating the toll band before it enters the crimping machine.","Products having 16 crimps per cm are described.","No information on the crimp ratio is provided.","It has now been unexpectedly discovered that hyper-fine crimping can be achieved effectively and without all-too drastic damage to the filaments if the total amount of the plasticizing components exceeds a value of 25 weight % at the time the crimping process occurs.","The concentrations in which the individual components are present does not matter.","The total content of the individual components is determined experimentally using a gravimetric method by taking a sample at 150.degree.","C. (20 min.)","directly downstream from the crimping machine.","As already noted, one of the solutions to the object of the invention relates to a filter tow that has a so-called mixed dernier.","DE 29 16 062 describes such a filter tow and the cigarette filters made from it.","In contrast to the filter tow of the invention, in this citation the most uniform possible distribution of the filaments is provided (see column 2, lines 22 ff.","in DE 29 16 062).","In the embodiment of DE 29 16 062, the effect of this is that the amount of material used in the filters that are to be produced is decreased with the same draw resistance, but as a consequence of the uniform distribution of the filaments the retentions in the filters increase significantly.","The object of the described invention is also to save material, but at the same time the retention in the filters made from the filter tow should be significantly less than that which is achieved when corresponding filters are made from a filter tow having a uniform filter dernier.","This is achieved when measures are taken to ensure that in the product of the invention the filaments of the coherent fiber bundles of the same filament dernier are formed of at least 600 filaments, preferably more than 1000 filaments, that differ in their filament demier from fiber bundle to fiber bundle by at least 1 dtex, preferably by more than 1.5 dtex.","The filter tow contains at least two, preferably more than five areas of filament bundles having the same filament dernier.","The advantage of this embodiment is that the filament bundles of the same filament dernier agglomerate in the filter and thus form areas of low, respectively high, draw resistance.","A filter manufactured from such a mixed-dernier tow accordingly has similar properties with respect to draw resistance and retention as those described, for example, in JP2000000085 for a classical core-jacket filter.","There are no restrictions with regard to crimp ratio and number of crimps per cm for this tow filter.","However, in a preferred embodiment of the invention, a product having a crimp ratio of more than 33%, preferably more than 35%, is made.","The advantages of the invention set forth in claims 8 to 10 may be summarized as follows: Based on the combination of materials set forth in the invention, working ranges with respect to draw resistance that are not currently available may be achieved.","By applying the teachings of claim 10 with the given draw resistance, the filament demier can be increased so that the products can achieve a significantly lower filtration performance.","As a result of the cited restriction with regard to the total demier, the consumption of material can be limited in manufacturing the filters.","However, a combination of the embodiments of at least one of claims 1 to 7 with one or more of claims 8 to 10 is especially advantageous.","This embodiment permits a further significant reduction of the total dernier to be used and, thus, results in further savings potential with regard to material consumption.","The invention is described in greater detail below based on the examples.","COMPARISON EXAMPLE (TOW OF TYPE 4.5 Y 55) A spinning solution having a concentration of 30 weight percent cellulose acetate in acetone is prepared, filtered, and then extruded through spinnerets.","The water content of the spinning solution is 1.5%.","The spinnerets are selected with regard to the number of holes, hole size, and number of spinnerets so that ultimately a tow of type 4.5 Y 55 is made.","The application of softening agent is set so that an oil coating of 0.5% results.","The tow band is crimped with a conventional stuffer box crimping machine so that after drying and packing, a filter tow having a crimp ratio of 30.5% results.","The total content of plasticizing components is 22.3%.","The tensile strength of the tow is 143 N. The number of crimps is 15.8 per cm filament length.","The product is processed into filter rods having a length of 126 mm and a diameter of 7.8 mm on a conventional filter rod machine, model KDF2/AF2, make: Hauni, Hamburg.","The generation of lint during the processing of the product is insignificant and amounts to 0.5 g/ton for the estimated consumption of one ton of material.","The performance curve of the product shown in FIG.","2 covers a draw resistance of 310-460 mm water column.","EXAMPLE 1 A spinning solution having a concentration of 30 weight parts cellulose acetate is prepared in acetone, filtered, and then extruded through spinnerets.","The water content of the spinning solution is 3%.","The spinnerets are selected with regard to number of holes, hole size, and number of spinnerets so that ultimately a filter tow of the type 4.5 Y 35 is produced.","The application of softening agent is adjusted so that an oil coating of 1.3% results.","The resulting tow band is crimped with a conventional stuffer box crimping machine so that, after drying and packaging, a filter tow having a crimp ratio of 43.5% results.","The total content of plasticizing components is 29%.","The tensile strength of the tow is 111 N. The number of crimps is 26 per cm filament length.","The product is processed into filter rods having a length of 126 mm and a diameter of 7.8 mm on a conventional filter rod machine, model KDF2/AF2, make: Hauni, Hamburg.","The generation of lint during the processing of the product is insignificant and amounts to 2.8 g/ton for the estimated consumption of one ton of material.","The performance curve of the product, shown in FIG.","2, covers a draw resistance range of 298-505 mm water column.","EXAMPLE 2 A spinning solution having a concentration of 30 weight parts cellulose acetate is prepared in acetone, filtered, and then extruded through spinnerets.","The water content of the spinning solution is 3%.","The spinnerets are selected with regard to number of holes, hole size, and number of spinnerets so that ultimately a filter tow of the type 4.5 Y 32 is produced.","Unlike in the above examples, the spinning machine is divided into six zones, and alternating zone by zone, a higher filament dernier (1185 filaments 6.9 den=7.65 dtex per zone) and a lower filament dernier (1185 filaments 2.1 den=2.3 dtex per zone) is spun.","The application of softening agent is adjusted so that an oil coating of 1.3% results.","The resulting tow band is crimped using a conventional stuffer box crimping machine so that after drying and packaging, a filter tow having a crimp ratio of 39.8% results.","The total content of plasticizing components is 27.5%.","The tensile strength of the tow is 129 N. The number of crimps is 23.1 per cm filament length.","The product is processed into filter rods having a length of 126 mm and a diameter of 7.8 mm on a conventional filter rod machine, model KDF2/AF2, make: Hauni.","The generation of lint during the processing of the product is insignificant and amounts to 0.9 g/ton for the estimated consumption of one ton of material.","The performance curve of the product covers a draw resistance range of 267-487 mm water column."]],"string":"[\n [\n \"FILTER TOW\",\n \"Abstract of the Disclosure The invention relates to a filter tow made from crimped endless cellulose acetate filaments, whereby the filaments form at least two connected fibre bundles with the same filament titre of at least 600 filaments.\",\n \"The filament titres differ from bundle to bundle by at least 1 dtex Furthermore a filter tow is disclosed with a crimp ratio of more than 40%, said filaments comprising at least 22 crimp turns per filament length.\"\n ],\n [\n \"1.A filter tow of crimped, continuous cellulose acetate filaments, wherein the filaments form at least two coherent fiber bundles of the same filament dernier of at least 600 filaments which differ from fiber bundle to fiber bundle in their filament dernier by at least 1 dtex.\",\n \"2.The filter tow of claim 1, wherein the filaments form more than five coherent fiber bundles of the same filament dernier.\",\n \"3.The filter of claim 1 or 2, wherein the individual fiber bundles contain more than 1000 filaments.\",\n \"4.The filter tow of one of claims 1 to 3, wherein the filament derniers of the filaments differ from fiber bundle to fiber bundle by more than 1.5 dtex.\",\n \"5.The filter tow of one of claims 1 to 4, wherein the crimp ratio is greater than 33%, and the number of crimps per cm is greater than 15.6.The filter tow of one of claims 1 to 5, wherein the crimp ratio is greater than 35% 7.The filter tow of one of the above claims, wherein the crimp ratio is greater than 40%, preferably greater than 42%, and the number of crimps per cm exceeds a value of 22.8.The filter tow of crimped, continuous cellulose acetate filaments, wherein the filter tow has a crimp ratio of greater than 40% and the filaments have at least 22 crimps per cm filament length.\",\n \"9.The filter tow of claim 8, wherein the crimp ratio is greater than 42% and the filaments have more than 25 crimps per cm.\",\n \"10.The filter tow of claim 8 or 9, wherein the filament dernier is greater than 3.3 dtex, and the total dernier is less than 44,000 dtex, preferably less than 40,000 dtex.\"\n ],\n [\n \"Detailed Description of the Invention DESCRIPTION The invention relates to a filter tow used in the manufacture of cigarette filters having a comparatively low filtration performance whose draw resistance retention characteristic was selected in such a way that their use in full-flavor cigarettes with a ventilation degree of more than 25% is possible while maintaining an acceptable draw resistance in the cigarette and an acceptable filter plug weight.\",\n \"The great majority of cigarette filters currently in use are manufactured from filter tow, which consists of continuous cellulose 2,5-acetate filaments.\",\n \"These filters are referred to as acetate filters.\",\n \"The cigarettes that are manufactured and sold in the European Union can be classified as follows based on their condensate contents in smoke: Full-flavor Medium Light, and Ultralight cigarettes Because of the maximum-content regulations (90/239/EEC) applying to condensate content, all of these cigarettes are ventilated.\",\n \"The degree of ventilation depends on the condensate target value and on the filtration performance of the filter being used.\",\n \"The following table summarizes the typical current values: [0009.1] Class mg Condensate Filament Dernier % Ventilation Full Flavor <12 2.8-3.3 dtex <25 Medium 8-10 ≦3.3 dtex <35 Light 4-8 2.1-2.8 dtex 30-50 Ultralight <41 1.7-8.8 >50 Most of products currently sold have a diameter of 7.8.+-.0.2 mm and a cigarette length of 85-120 mm.\",\n \"In addition, there are niche products whose design characteristics differ significantly from the most commonly sold products.\",\n \"In response to this wide variety of product requirements, the filter tow industry supplies a multitude of different tow types that can essentially be described by the following characteristics: filament dernier (dtex) 1.6-8.8; total dernier (dtex) 17,000-60,000; crimp index (%) 25 max.\",\n \"38; number of crimps per cm: 10 max.\",\n \"20.The following description of the object of the invention exclusively relates to the class of full-flavor products.\",\n \"A typical filter cigarette of the full-flavor class has the following typical characteristics (these values are approximate typical values).\",\n \"They serve simply as a basis for calculations for further embodiments.\",\n \"Lengths: Cigarette:84 mm Filter:24 mm Tobacco rod:63 mm Tipping:26 mm Diameters: Filter:7.80 mm Cigarette:7.85 mm Draw resistance: Filter:66 mm water column Tobacco rod:50 mm water column Cigarette:100 mm water column Filter tow specification: 3.0 Y 35 (filament dernier: 3.33 dtex; total dernier: 38.900, cross-sectional shape of the filaments: Y) Tobacco mixture: American Blend Ventilation: Degree of ventilation:20% Position of ventilation zone:11 mm from the mouth end Smoke values: Condensate:12 mg Nicotine:0.9 mg Carbon monoxide (CO): 14 mg Retention values: Condensate:42% Nicotine:38% Carbon monoxide (CO):None As a consequence of the new maximum-content regulation (2000/C 150E/03) of the European Union, full-flavor products in particular will have to undergo drastic changes in their design features, since effective Jan. 1, 2004 not only the condensate value, but also the nicotine value--and in particular the CO value--will be regulated with respect to their maximum limits.\",\n \"The challenge involving the modification of the products to meet the new EU regulations will be that the permitted maximum amounts with an optimal smoke flavor will need to hit the values 10 mg condensate, 1 mg nicotine, and 10 mg CO per cigarette as precisely as possible.\",\n \"In other words, in the future a 1:1 ratio of CO/tar must be targeted for full-flavor products.\",\n \"As the above table shows, the ratio in this example is 1.17.Based on the model calculations that solely take a change in filter ventilation into account, it can be assumed that a filter ventilation of approximately 35% will be required to reduce the CO from the current value in the above calculation example of 14 mg to a future value of 10 mg per cigarette.\",\n \"An increase in the degree of ventilation in an otherwise unchanged product will unavoidably necessitate a decrease in the cigarette draw resistance.\",\n \"For consumers however, an optimal smoke flavor means that the draw resistance of the cigarette must not fall significantly below the current value of approximately 100 mm water column.\",\n \"Increasing the degree of ventilation, however, will mean that maintaining the other filter parameters such as draw resistance, diameter, position of the ventilation zone, and the filter length will significantly reduce the draw resistance of the cigarette.\",\n \"The resulting cigarette draw resistance can be calculated using the following equation, which is based on a laminar flow model: Pv=Po-Vx(Po-PfxLv/Lf), where: Pv: Draw resistance of the ventilated cigarette (mm water column) Po: Draw resistance of unventilated cigarette: =draw resistance [filter+tobacco rod] (mm water column) Pf: Filter draw resistance (mm water column) V: Ventilation degree of filter (%)/100 Lv: Distance of the ventilation zone from the mouth end (mm) Lf: Filter length Thus, the open draw resistance will then only be 88 mm water column (instead of the original 100 mm water column) for the stated sample cigarette at a ventilation degree of 35%.\",\n \"Such a drop in the cigarette draw resistance is clearly perceivable for the smoker and it must be offset by an increase in the filter resistance (in the above example to approximately 80 mm water column) in order to maintain the consumer's smoke feel.\",\n \"This increase in the filter draw resistance results in an unavoidable increase in the CO/tar ratio, since the tar values, but not the CO values, are influenced by the filter draw resistance (an increase in the filter draw resistance results in a decrease in the tar values).\",\n \"Therefore, the draw resistance/retention ratios must be redefined for future products in the \\\"full-flavor\\\" category.\",\n \"The adjustment to a 1:1 CO/tar ratio and the filter tow used to manufacture the filter needed to achieve an acceptable product with regard to cigarette draw resistance must be modified to meet these requirements.\",\n \"In this regard, the following specifications may be defined: 1.The filter tow must be suitable for producing the draw resistance values that are typical today (however preferably even higher values).\",\n \"2.The filtration performance of the filter made with the filter tow of the invention must, with the given filter draw resistance, be significantly lower than the current typical retention values in order to achieve the 1:1 CO/tar ratio.\",\n \"3.The additional material consumption that results from the increase of the filament dernier must be kept within acceptable limits.\",\n \"With a given filter diameter and length, the retention of a filter depends exclusively on the draw resistance and the filament dernier.\",\n \"For example, an increase in the filament dernier and a decrease in the draw resistance results in a drop in filtration performance (and vice-versa).\",\n \"Since the interrelationships are of a very complex nature, filter calculation programs are used to estimate the extent of such changes.\",\n \"One example of such calculation programs is Cabled (Capability Line Expert; copyright Rhodia Acetow GmbH 1998, Freiburg, Germany) from the firm of Rhodia Acetow.\",\n \"In the prior art, therefore, the retention of a filter is reduced by increasing the filament dernier.\",\n \"Model calculations suggest that in the future the filament dernier will need to be increased by at least 0.5-1.5 dtex in order to meet the requirements for filtration performance.\",\n \"However, the filter tow specifications commonly encountered in the market (the mean total dernier of all specifications is approximately 35 ktex) are not suitable for also covering the required draw resistance range with the required minimum filament dernier (retention) (Cable.COPYRGT.\",\n \"calculation program from Rhodia Acetow GmbH).\",\n \"In the prior art, this problem can only be solved with a monoacetate filter by drastically increasing the filter dernier to well above 45,000 dtex.\",\n \"Such solutions, for example a filter tow of the type 4.5 Y 55 (see FIG.\",\n \"2) in the prior art would then meet the draw resistance and retention specifications, but would result in an increase in the consumption of filter tow of >35% compared with current products.\",\n \"The above discussion was limited to a description of the full-flavor group of cigarettes.\",\n \"If, however, a CO/tar ratio of 1:1 is generally sought, the discussion regarding the widening of the filter resistance ranges would apply to all filament derniers.\",\n \"This means that the object of the invention described below will generally comprise the currently standard filament dernier spectrum of 1.6-8.8 dtex.\",\n \"Therefore, the object of the invention is to disclose a filter tow with which it is possible to manufacture filters that have a filtration performance achieving a CO/tar ratio of 1:1.In order to meet the above requirements, the invention proposes two approaches that are basically independent of each other and whose elements can, if desired, be combined.\",\n \"The solution of the invention relates to a filter tow of crimped, continuous cellulose acetate filaments, in which the filaments form at least two coherent fiber bundles that have the same filament dernier and have at least 600 filaments and that differ in their filament dernier from fiber bundle to fiber bundle by at least 1 dtex.\",\n \"In particular, this solution produces outstanding results with regard to filtration performance, since it was unexpectedly found that a filter that is manufactured from such a non-homogeneously distributed mixed tow exhibits a significantly lower tar retention than does a product made of a conventional tow with a uniform filament dernier distribution or a mixed tow having a homogeneous filament distribution (see FIG.\",\n \"3).\",\n \"The second solution is accomplished by a filter tow that has a more intensive and at the same time markedly finer crimp structure than the typical products encountered today.\",\n \"Claim 4 recites a filter tow having a crimp index of more than 40%, in which the filaments have at least 22 curls per cm filament length.\",\n \"In particular, this filter tow fulfills the maximum requirements with regard to filter draw resistance.\",\n \"The values that are achieved are even exceeded if the crimp index is more than 42%, whereby the filaments preferably have more than 25 curls per cm.\",\n \"In order at the same time to achieve optimal filtration performance by the filters, a filter tow having a filament dernier of more than 3.33 dtex and a total dernier of less than 44,000 dtex, preferably less than 40,000 dtex is proposed.\",\n \"The limitation on the total dernier results in an increased filter tow consumption that is still acceptable.\",\n \"The design and the advantages of the two solutions of the invention may be described as follows: The crimp index Ix is a measure of the intensity of curling.\",\n \"The crimp index of a filter tow is determined by means of the tensile test (load/elongation relationship).\",\n \"It is defined as the ratio of the elongated length L2 under the test load minus the initial length over the initial length under preload L1: Ix=(L2-L1)/L1*100(%) Here, the test load is 25 N and the preload is 2.5 N. The length between the jaws of the tensile test machine is 250 mm.\",\n \"The curling index is determined in a tensile test at a constant traverse rate of 300 mm/min on a G02 test machine manufactured by Borgwaldt GmbH, Hamburg.\",\n \"10 individual readings were taken per measurement.\",\n \"Testing is performed in a standard climate: 20.degree.\",\n \"C. and 60% relative humidity.\",\n \"(The test method for determining the crimp ratio is described in the Cable program under the \\\"Help\\\" button).\",\n \"The crimp ratio values of current filter tow specifications normally lie between 25 and 35%.\",\n \"The firm of Rhodia Acetow is the sole manufacturer that offers worldwide specifications under the designation SK in a filament dernier range of <2.78 dtex that covers a crimp ratio range of from greater than 35% to a maximum of 38%.\",\n \"As already stated, the crimp ratio is a measure of the intensity of crimping and, thus, the position of the performance curve.\",\n \"In this context, a performance curve is understood to mean a performance range of a filter tow, which states the draw resistance range in which a filter tow can be processed with a given diameter and filter rod length under typical processing conditions.\",\n \"For this working range, the performance curve describes the relationship between the draw resistance and weight.\",\n \"The relationship between the position of the performance curve and the crimp ratio is shown using the example of a filter tow of type 2.1 Y 35 HK (filament dernier: 2.33 dtex; total demier: 38,900; cross-sectional shape of the filaments: Y; crimp ratio: 32%) and a 2.1 Y 35 SK (filament dernier: 2.33 dtex; total demier: 38,900, cross-sectional shape of the filament: Y; crimp ratio: 36%) in FIG.\",\n \"1.The above object is accomplished by manufacturing a filter tow that has a crimp ratio of more than 40%, preferably more than 42%.\",\n \"This ratio is absolutely necessary in order to cover the corresponding draw resistance range.\",\n \"The crimp ratio is affected by the frequency and the amplitude of the crimps.\",\n \"The number of crimps provides a measure of the fineness of crimping.\",\n \"Currently, typical values range from 10 to max.\",\n \"20 crimps per cm filament length.\",\n \"The SK tows referred to above normally have 15-18 crimps per cm.\",\n \"It was unexpectedly discovered that such an index can only be achieved with an acceptable filter tow tensile strength if the crimping is hyper-fine.\",\n \"Therefore, the filter tow of the invention has, in addition to an index of more than 40% in the filaments, on average more than 22 crimps per cm, preferably more than 25.Such a filter tow can be processed on conventional filter rod machines into cigarette filters that have a draw resistance level comparable to filters manufactured using the currently typical specifications.\",\n \"FIG.\",\n \"2 shows the working range of a tow of type 4.5 Y 35 of the invention (filament dernier: 5.0 dtex; total demier: 38,900 dtex) in comparison with a conventional filter tow of type 3.0 Y 35 (filament dernier 3.33 dtex: total dernier: 38,900 dtex) respectively a conventional 4.5 Y 55 and a conventional 4.5 Y 35.FIG.\",\n \"2 clearly shows that, compared with the current situation (3.0 Y 35) significantly more material is consumed with the solution of the invention (Example 1: 4.5 Y 35) as a result of the increase of the filament dernier, but that this additional consumption ends up being significantly less than with a traditional prior-art solution (4.5 Y 55).\",\n \"In order to produce hyper-fine crimping without being overly aggressive, certain basic conditions must be met in the manufacturing process.\",\n \"Prior to the crimping process, the filter tow band contains the following components that soften the acetate filaments: water, acetone, and softening components.\",\n \"It is known that these softening components have a lasting effect on the crimping process.\",\n \"For example, U.S. Pat.\",\n \"No.\",\n \"5,225,277 describes the effect of acetone and tow band temperatures on the stability of crimping.\",\n \"Products having up to 19.7 crimps per cm can be manufactured using the method described therein.\",\n \"This document does not make any statements with regard to the crimp ratio.\",\n \"However, using the data stated therein (draw resistance, tow specification and machine settings) a crimp ratio that is clearly below 35% can be estimated with the help of the Cable software referred to above.\",\n \"JP unexamined patent document 56-53223 describes the effects of steam-treating the toll band before it enters the crimping machine.\",\n \"Products having 16 crimps per cm are described.\",\n \"No information on the crimp ratio is provided.\",\n \"It has now been unexpectedly discovered that hyper-fine crimping can be achieved effectively and without all-too drastic damage to the filaments if the total amount of the plasticizing components exceeds a value of 25 weight % at the time the crimping process occurs.\",\n \"The concentrations in which the individual components are present does not matter.\",\n \"The total content of the individual components is determined experimentally using a gravimetric method by taking a sample at 150.degree.\",\n \"C. (20 min.)\",\n \"directly downstream from the crimping machine.\",\n \"As already noted, one of the solutions to the object of the invention relates to a filter tow that has a so-called mixed dernier.\",\n \"DE 29 16 062 describes such a filter tow and the cigarette filters made from it.\",\n \"In contrast to the filter tow of the invention, in this citation the most uniform possible distribution of the filaments is provided (see column 2, lines 22 ff.\",\n \"in DE 29 16 062).\",\n \"In the embodiment of DE 29 16 062, the effect of this is that the amount of material used in the filters that are to be produced is decreased with the same draw resistance, but as a consequence of the uniform distribution of the filaments the retentions in the filters increase significantly.\",\n \"The object of the described invention is also to save material, but at the same time the retention in the filters made from the filter tow should be significantly less than that which is achieved when corresponding filters are made from a filter tow having a uniform filter dernier.\",\n \"This is achieved when measures are taken to ensure that in the product of the invention the filaments of the coherent fiber bundles of the same filament dernier are formed of at least 600 filaments, preferably more than 1000 filaments, that differ in their filament demier from fiber bundle to fiber bundle by at least 1 dtex, preferably by more than 1.5 dtex.\",\n \"The filter tow contains at least two, preferably more than five areas of filament bundles having the same filament dernier.\",\n \"The advantage of this embodiment is that the filament bundles of the same filament dernier agglomerate in the filter and thus form areas of low, respectively high, draw resistance.\",\n \"A filter manufactured from such a mixed-dernier tow accordingly has similar properties with respect to draw resistance and retention as those described, for example, in JP2000000085 for a classical core-jacket filter.\",\n \"There are no restrictions with regard to crimp ratio and number of crimps per cm for this tow filter.\",\n \"However, in a preferred embodiment of the invention, a product having a crimp ratio of more than 33%, preferably more than 35%, is made.\",\n \"The advantages of the invention set forth in claims 8 to 10 may be summarized as follows: Based on the combination of materials set forth in the invention, working ranges with respect to draw resistance that are not currently available may be achieved.\",\n \"By applying the teachings of claim 10 with the given draw resistance, the filament demier can be increased so that the products can achieve a significantly lower filtration performance.\",\n \"As a result of the cited restriction with regard to the total demier, the consumption of material can be limited in manufacturing the filters.\",\n \"However, a combination of the embodiments of at least one of claims 1 to 7 with one or more of claims 8 to 10 is especially advantageous.\",\n \"This embodiment permits a further significant reduction of the total dernier to be used and, thus, results in further savings potential with regard to material consumption.\",\n \"The invention is described in greater detail below based on the examples.\",\n \"COMPARISON EXAMPLE (TOW OF TYPE 4.5 Y 55) A spinning solution having a concentration of 30 weight percent cellulose acetate in acetone is prepared, filtered, and then extruded through spinnerets.\",\n \"The water content of the spinning solution is 1.5%.\",\n \"The spinnerets are selected with regard to the number of holes, hole size, and number of spinnerets so that ultimately a tow of type 4.5 Y 55 is made.\",\n \"The application of softening agent is set so that an oil coating of 0.5% results.\",\n \"The tow band is crimped with a conventional stuffer box crimping machine so that after drying and packing, a filter tow having a crimp ratio of 30.5% results.\",\n \"The total content of plasticizing components is 22.3%.\",\n \"The tensile strength of the tow is 143 N. The number of crimps is 15.8 per cm filament length.\",\n \"The product is processed into filter rods having a length of 126 mm and a diameter of 7.8 mm on a conventional filter rod machine, model KDF2/AF2, make: Hauni, Hamburg.\",\n \"The generation of lint during the processing of the product is insignificant and amounts to 0.5 g/ton for the estimated consumption of one ton of material.\",\n \"The performance curve of the product shown in FIG.\",\n \"2 covers a draw resistance of 310-460 mm water column.\",\n \"EXAMPLE 1 A spinning solution having a concentration of 30 weight parts cellulose acetate is prepared in acetone, filtered, and then extruded through spinnerets.\",\n \"The water content of the spinning solution is 3%.\",\n \"The spinnerets are selected with regard to number of holes, hole size, and number of spinnerets so that ultimately a filter tow of the type 4.5 Y 35 is produced.\",\n \"The application of softening agent is adjusted so that an oil coating of 1.3% results.\",\n \"The resulting tow band is crimped with a conventional stuffer box crimping machine so that, after drying and packaging, a filter tow having a crimp ratio of 43.5% results.\",\n \"The total content of plasticizing components is 29%.\",\n \"The tensile strength of the tow is 111 N. The number of crimps is 26 per cm filament length.\",\n \"The product is processed into filter rods having a length of 126 mm and a diameter of 7.8 mm on a conventional filter rod machine, model KDF2/AF2, make: Hauni, Hamburg.\",\n \"The generation of lint during the processing of the product is insignificant and amounts to 2.8 g/ton for the estimated consumption of one ton of material.\",\n \"The performance curve of the product, shown in FIG.\",\n \"2, covers a draw resistance range of 298-505 mm water column.\",\n \"EXAMPLE 2 A spinning solution having a concentration of 30 weight parts cellulose acetate is prepared in acetone, filtered, and then extruded through spinnerets.\",\n \"The water content of the spinning solution is 3%.\",\n \"The spinnerets are selected with regard to number of holes, hole size, and number of spinnerets so that ultimately a filter tow of the type 4.5 Y 32 is produced.\",\n \"Unlike in the above examples, the spinning machine is divided into six zones, and alternating zone by zone, a higher filament dernier (1185 filaments 6.9 den=7.65 dtex per zone) and a lower filament dernier (1185 filaments 2.1 den=2.3 dtex per zone) is spun.\",\n \"The application of softening agent is adjusted so that an oil coating of 1.3% results.\",\n \"The resulting tow band is crimped using a conventional stuffer box crimping machine so that after drying and packaging, a filter tow having a crimp ratio of 39.8% results.\",\n \"The total content of plasticizing components is 27.5%.\",\n \"The tensile strength of the tow is 129 N. The number of crimps is 23.1 per cm filament length.\",\n \"The product is processed into filter rods having a length of 126 mm and a diameter of 7.8 mm on a conventional filter rod machine, model KDF2/AF2, make: Hauni.\",\n \"The generation of lint during the processing of the product is insignificant and amounts to 0.9 g/ton for the estimated consumption of one ton of material.\",\n \"The performance curve of the product covers a draw resistance range of 267-487 mm water column.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476920"}}},{"rowIdx":983,"cells":{"text":{"kind":"list like","value":[["Liquid flavor beverage system","A beverage apparatus (30) having a plurality of dispensing modules (32) and a housing (34) that defines a chamber (36) containing water or liquide suitable for use in brewing."],["1.A liquid flavor dispensing system for use with a beverage making apparatus, the liquid flavor dispensing system comprising: a container for retaining a liquid flavoring; a passage connected to the container and extending from the container to a portion of a beverage making apparatus; a pump positioned along the passage for moving liquid flavor from the container to a portion of the beverage making apparatus; and means for controlling flow from the passage, the flow control means communicating with the passage for controlling flow therefrom.","2.The liquid flavor dispensing system as in claim 1, further comprising means for monitoring a quantity of liquid flavor in the container.","3.The liquid flavor dispensing system as in claim 2, wherein the monitoring means includes a sensor associated with the container for detecting the quantity of liquid flavor in the container.","4.The liquid flavor dispensing system as in claim 1, wherein the monitoring means includes a sensor associated with the pump for detecting a characteristic of the pump for inferring the quantity of liquid flavor in the container.","5.A beverage making apparatus comprising: a liquid distribution system; means for combining a beverage making substance with a liquid communicating with the liquid distribution system; a liquid flavor dispensing system for dispensing liquid flavor to at least one of the liquid distribution system and the combining means.","6.The beverage making apparatus as in claim 5, further comprising the combining means including a beverage brewing portion for combining liquid from the liquid distribution system with a beverage brewing substance retained in the beverage brewing portion.","7.The beverage making apparatus of claim 5, further comprising the combining means including a powdered beverage mixing portion for combining liquid from the liquid distribution system with a powdered beverage substance.","8.The beverage making apparatus of claim 5, further comprising the combining means including a liquid beverage concentrate mixing portion for combining liquid from the liquid distribution system with a liquid beverage concentrate.","9.The beverage making apparatus of claim 5, further comprising: the liquid distribution system having an outlet proximate to the combining means; and the liquid flavor dispensing system dispensing liquid flavor to the outlet of the liquid distribution system prior to combining with a beverage making substance.","10.The beverage making apparatus of claim 5, further comprising: the liquid distribution system having a dispensing line for delivering liquid to the combining means; and The liquid flavor dispensing system dispensing liquid flavor to the dispensing line for mixing flavor with the liquid prior to mixing the liquid with the beverage making substance.","11.The beverage making apparatus of claim 5, further comprising: the liquid flavor dispensing system dispensing liquid flavor to the combining means for dispensing flavor into the beverage making substance.","12.A method of combining liquid flavor for use with a beverage making apparatus, the method comprising the steps of: providing a liquid flavor; providing a passage for dispensing the liquid flavor to a beverage making apparatus; dispensing a quantity of liquid flavor through the passage; and combining the dispensed liquid flavor with a beverage for modifying the flavor of the beverage.","13.The method of combining a liquid flavor for use with a beverage making apparatus of claim 12, further comprising the steps of: providing a liquid distribution system in combination with the beverage making apparatus; and dispensing liquid flavor to a location proximate to the dispensing of liquid from the liquid distribution system for combining the liquid flavor with the liquid prior to combining with the beverage making substance.","14.The method of combining a liquid flavor for use with a beverage making apparatus of claim 12, further comprising the steps of: providing a liquid distribution system having a dispensing line for delivering liquid to combine with a beverage making substance; and dispensing the liquid flavor to the dispensing line for mixing the flavor with the liquid prior to mixing the liquid with the beverage making substance.","15.The method of combining a liquid flavor for use with a beverage making apparatus of claim 12, further comprising the step of: dispensing liquid flavor to the beverage making substance."],["<SOH> BACKGROUND <EOH>This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.","Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.","In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.","Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.","As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.","Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.","Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.","Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.","These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.","To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.","In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process."],["<SOH> SUMMARY <EOH>The present disclosure relates to apparatus for making and dispensing beverages.","The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.","Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.","The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.","In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.","In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.","The apparatus may also include a plurality of conduits for supplying liquid to the assembly.","The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.","Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.","The housing may be positioned behind the beverage dispensing modules.","In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.","With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.","The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.","A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.","The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.","The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.","In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.","In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.","Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived."],["BACKGROUND This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.","Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.","In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.","Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.","As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.","Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.","Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.","Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.","These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.","To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.","In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process.","SUMMARY The present disclosure relates to apparatus for making and dispensing beverages.","The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.","Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.","The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.","In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.","In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.","The apparatus may also include a plurality of conduits for supplying liquid to the assembly.","The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.","Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.","The housing may be positioned behind the beverage dispensing modules.","In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.","With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.","The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.","A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.","The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.","The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.","In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.","In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.","Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived.","BRIEF DESCRIPTION OF THE DRAWINGS The detailed description particularly refers to the accompanying figures in which: FIG.","1 is a partial fragmentary side elevational view of a modular beverage dispensing apparatus; FIG.","2 is a top plan view of the modular beverage dispensing apparatus of FIG.","1; FIG.","3 is a side elevational view of the modular beverage dispensing apparatus of FIGS.","1 and 2; FIG.","4 is a schematic illustration of the modular beverage dispensing apparatus; FIG.","5 is a schematic illustration of one module of the modular beverage dispensing apparatus; FIG.","6 is a diagrammatic illustration of a dispensing and disposal valve configuration; FIG.","7 is a partial fragmentary top plan view taken along 7-7 in FIG.","6; FIG.","8 is a diagrammatic illustration of a liquid flavor distributing system; FIGS.","9 and 10 are diagrammatic illustrations of the arrangement of liquid flavor distributing systems; FIG.","11 is a cross sectional view, schematic in nature, of a portion of a powdered beverage, soluble beverage dispensing system including connections to dispense liquid flavor to the soluble beverage production stream; FIGS.","12 and 13 are diagrammatic illustrations of the liquid flavor distributing system coupled to the soluble beverage dispensing system of FIG.","11; FIG.","14 is a perspective view of a soluble beverage dispenser having a slide-out assembly; FIG.","15 is an enlarged perspective view of the soluble beverage dispenser of FIG.","14 showing a user removing a container from the soluble beverage dispensing device using the slide-out assembly; FIGS.","16 and 17 are enlarged views of portions of the slide-out assembly of the soluble beverage dispenser of FIG.","14; FIG.","18 is a perspective view of a cable and spring leverage system which assists movement of the slide-out assembly relative to the beverage dispensing system; FIGS.","19 and 20 are schematic illustrations of a communication link between a base station and a beverage dispensing device; and FIGS.","21-28 are schematic illustrations of the circuitry associated with the communication system of FIGS.","19 and 20 AND INCLUDE FIG.","22A, FIGS.","22-22D, FIGS.","23-23D, FIG.","24, FIGS.","24A-24B, FIGS.","25A-25B, FIGS.","26A-26, FIG.","27, FIGS.","27A-27C, FIG.","28 and FIGS.","28A-28E.","DESCRIPTION OF THE DRAWINGS While the present disclosure may be susceptible to embodiment in different forms, there is shown in the drawings, and herein will be described in detail, embodiments with the understanding that the present description is to be considered an exemplification of the principles of the disclosure and is not intended to limit the disclosure to the details of construction and the arrangements of components set forth in the following description or illustrated in the drawings.","A beverage dispensing apparatus 30 illustrated generally in FIGS.","1-4 as comprising a plurality of beverage dispensing modules 32 and a housing 34 that defines a chamber 36 containing water or other liquid suitable for use in brewing.","The housing 34 desirably acts as a central reservoir for a constant supply of heated water to be used by the beverage dispensing modules 32 to brew several different beverages.","The beverage dispensing apparatus 30 may have any suitable construction and configuration.","The beverage apparatus 30 may include any suitable number of beverage dispensing modules 32 oriented in any suitable manner, depending on various circumstances, such as, for example, customer demand, customer usage, available store space, and desired aesthetics.","Similarly, the housing 34 may have any suitable construction and configuration that desirably is consistent with customer demand, customer usage, available store space and desired aesthetics.","A plurality of conduits in the form of water lines 38 provide flow communication between the housing 34 and the beverage dispensing modules 32 to supply water for brewing.","In a preferred embodiment, the housing 34 is positioned behind the beverage dispensing modules 32 and centrally located relative to the beverage dispensing modules 32 to minimize the distance between the housing and each of the beverage dispensing modules, thereby increasing operational efficiency and minimizing space.","Terms including beverage and beverage making as used herein are intended to be broadly defined as including but not limited to the brewing of coffee, tea and any other brewed beverage, including a final beverage or food product as well as producing an intermediate product to be combined with a final beverage or food product.","This broad interpretation is also intended to include, but is not limited to any process of infusing, steeping, reconstituting, diluting, dissolving, saturating or passing a liquid through or otherwise mixing or combining a beverage substance with a liquid such as water without a limitation to the temperature of such liquid unless specified.","This broad interpretation is also intended to include, but is not limited to beverage substances such as ground coffee, tea, liquid beverage concentrate, powdered beverage concentrate, freeze dried coffee or other beverage concentrates, to obtain a desired beverage or other food.","In the illustrated embodiment, for example, the beverage dispensing apparatus 30 comprises ten beverage dispensing modules 32 oriented in a generally U-shaped manner that substantially surrounds the housing 34.As illustrated, four of the beverage dispensing modules 32 face the same direction, four other beverage dispensing modules 32 face an opposite direction, and two of the beverage dispensing modules 32 are positioned in a space-efficient manner consistent with a desired orientation.","A simplified form of the beverage apparatus 30 is shown in FIG.","4 which uses simplified block diagram illustrations to show multiple beverage dispensing modules 32 which are configured to provide a beverage brewing system.","The beverage apparatus 30 shown in FIG.","4 is very similar to a soluble beverage apparatus 30a also shown in FIG.","4.The beverage apparatus 30, as will be described in greater detail herein below, relates to a beverage brewing or producing system whereas the soluble beverage apparatus 30a, as will be described in greater detail herein below, relates to a soluble or powdered beverage dispensing system.","Both beverage apparatuses 30, 30a employ a common heated water reservoir contained in housing 34 to which the individual modules 32, 32a are connected.","As shown in FIGS.","2-4, the housing 34 desirably is centrally located to serve multiple beverage modules 32, 32a.","The benefit of a common housing 34 functioning as a common reservoir to supply water, instead of individual tanks for each module 32, 32a, is that it substantially reduces the cost and greatly increases the efficiency of the system.","Also it reduces the redundancy of multiple heated water reservoirs, reduces the wattage required by the system and increases flexibility in the number of modules 32, 32a which might be included.","The particular configuration with the housing 34 positioned in close proximity to or between several modules 32, 32a is beneficial in that it also minimizes the length of water lines 38 connecting the housing 34 to the modules 32, 32a.","Additionally, a larger body of water in the housing 34 tends to help maintain the temperature and reduce temperature loss over time as well as maintaining a larger volume of brew water at a desired temperature.","While the housing 34 retained within the beverage apparatus 30 is insulated to reduce heat loss and reduce heat load on the ambient environment, heat which may escape from the housing 34 will generally pass to the modules 32, thereby increasing the efficiency of maintaining the temperature of the beverages retained in the modules 32.Insulation desirably is provided to reduce beat loss to the ambient environment.","This will also help reduce the exertion on the ambient air-conditioning system thereby further increasing efficiency of the present beverage apparatus 30, 30a.","With reference to FIGS.","1-4, each module 32, 32a produces a separate brewed beverage.","The centralization of a housing 34 helps reduce the individual cost associated with the module 32,32a and improves the ease of manufacturing of the modules.","Also, there is increased commonality of parts associated with this type of system.","The modular configuration of the beverage apparatus 30 using the modules 32, 32a increases the potential variability of the physical size and brewer configuration of the system.","This is beneficial to accommodate the needs of a particular location.","In other words, only a desired or required number of modules 32, 32a will be needed at any given time.","If a location has an increase in demand for brewed or soluble beverages additional modules 32, 32a may be added as necessary for the particular change in demand.","Additionally, with regard to repair and maintenance, the beverage apparatus 30, 30a can be designed such that individual modules 32, 32a can be removed for repair and maintenance as necessary with an identical module being inserted in the missing modules.","In a preferred embodiment, (FIG.","5) the housing 34 includes a heating element 52, and one or more system sensors such as a level sensor 86 and a temperature sensor 84.Such components tend to be generally readily available and easily replaced and as such further enhance the reliability of the overall beverage apparatus 30, 30a.","With reference to FIG.","3, the line 38 connecting each housing 34 to a brewing portion 40 of the module 32 is shown.","Water 42 retained in the housing 34 can be readily dispensed.","A water inlet line 44 is provided to introduce unheated water 50 to the housing 34.The inlet line 44 runs from an upper portion 46 of the housing 34 and directs the inward water downwardly towards a bottom portion 48 of the housing 34.Introducing the unheated water 50 towards the bottom of the reservoir introduces the unheated water 50 in close relation to a heated element 52 thereby accelerating the heating of the water.","Additionally, the introduction of unheated water 50 in the bottom portion 48 tends to force heated water in the upper portion 46 upwardly.","As such, the water 42 in the upper portion 46 tends to be the hottest water in the housing 34.Introducing unheated water 50 from the upper portion has the benefit of providing an entrance into the reservoir through a reservoir wall 56 with little or no pressure on the entry therethrough.","This may result in an increase in the reliability of the entry therethrough.","Additionally, a reinforcing or strengthening structure 58 is provided extending between opposed walls 60, 62 of the housing 34.This helps prevent bowing or stress on the walls 60, 62 which might otherwise result from the force of the water on the walls.","Multiple reinforcing structures 58 may be provided throughout the housing 34 as necessary to provide support for the size and dimension of the housing required for a particular system.","FIG.","5, for example, shows a diagrammatic illustration of an embodiment of the apparatus 30 shown in FIGS.","1-4.The beverage dispensing modules 32 can have any suitable construction desirably adapted to produce beverages, store beverages for ready consumption, and facilitate ready dispensing of the beverages.","Each module 32 includes a brewing assembly 30 that may have any suitable construction that can be used to produce beverage such as brewing coffee or other beverages.","The illustrated brewing assembly 30, for example, includes a brewing substance retainer or assembly 64 including a funnel 66.Such a retainer 64 may be well known in the art and may include a filter (not shown) retained in the funnel 66 for holding a charge of beverage substance.","The funnel 66 may be positioned above a reservoir or container portion 68 of the dispensing module 32 which receives brewed beverage from the funnel.","A dispensing assembly 70 in the form of a controllable valve may be associated with the container 68 for dispensing beverage from the module 32.With reference to FIG.","5 a simplified review of the beverage brewing process is described.","A brew actuator switch 72 desirably is associated with each of the modules 32.To initiate a brew cycle for one of the modules 32 a respective brew activation switch 72 is operated.","Operation of the switch 72 sends a signal over line 74 to controller 76.Controller 76 operates an inlet valve 78 over line 80 to introduce water to the housing 34.Also, heating element 52 is coupled to the controller 76 over line 82.The temperature sensor 84 and level sensor 86 are coupled to the controller over lines 88, 90 respectively.","If the controller 76 detects proper level and temperature it will initiate a brew cycle thereby displacing water from the housing 34 through a sprayhead assembly 92.The sprayhead assembly 92 communicates with the substance retainer 64 and dispenses water into the funnel 66 for producing a beverage therein.","The illustrated module 32 includes a level sensor 94 associated with the container 68 which is coupled to the control over line 96.The level sensor can detect a change in level in beverage retained in the container 68 which may be used to activate a brew cycle.","Also, the level detector 94 may sense the level in the beverage container 68 thereby indicating if the container is full or above the desired level to prevent initiation of a brew cycle.","This will prevent over flow of the beverage server 68.The beverage dispensing modules 32 can be constructed to dispense beverage in any suitable manner.","In a preferred embodiment, for example, a dispenser or faucet switch 98 may be associated with the dispensing assembly 70 and coupled with a dispensing handle or control 100.Activation of the control 100 will activate the switch 98 over line 102 connected to the controller 76 thereby indicating that the controller 76 should operate a solenoid valve 104 (FIGS.","6 and 7), via line 106 to dispense beverage therefrom.","As will be described in greater detail below with regard to FIGS.","6 and 7, a disposal or dump valve 108 is also coupled with the container 68 to facilitate controllable disposal, via line 110 coupled to the controller 76.With reference to FIGS.","6 and 7, the beverage container 68 may further include a manifold 114.The illustrated manifold 114 includes a primary passage 116 which communicates with an interior cavity or reservoir 118 of the container 68.Beverage flows from the cavity 118 through the primary passage 116.The solenoid valves 104, 108 communicate with the passage 116.A dispensing passage 120 and a disposal passage 122 communicate with the primary passage 116.Solenoid valve 104 and 108 desirably are normally closed and require specific activation in order to open the valves.","During a dispense cycle the switch or control 100, for example a controllable faucet as shown in FIG.","5, is activated thereby activating solenoid valve 104.When the dispense solenoid valve 104 is opened beverage 124 will flow from the cavity through the solenoid valve 104.Under this condition disposal valve 108 is closed.","When beverage must be disposed of, the disposal solenoid valve 108 is opened while maintaining the dispense solenoid valve 104 in a closed position.","This will allow the disposal of beverage through a separate dispensing line.","Disposal of beverage may occur as a result of a time lapsed period whereby after a specified period of time beverage should be disposed of.","For example, if a beverage only maintains a desired flavor profile or other characteristics for a period of time, for example 2 hours, at the end of two hours after the beverage brewing cycle is initiated, the beverage may be disposed of through the disposal valve 108.Such disposal may also be used at the end of a service cycle such that at the end of the day all beverages may be disposed of at a predetermined time.","The disposal valve may also be used in a manner to flush and cleanse the beverage brewing system.","In this regard, at a predetermined period of time, the controller 76 may operate the disposal valve 108 to dispose of any beverage retained in the container 68.The disposal valve 108 and dispense valve 104 may be closed.","The controller 76 may then dispense a quantity of water into the cavity 118 thereby flushing and sanitizing the reservoir.","After holding the heated water for a predetermined period of time the water may be disposed through the disposal valve 108.Multiple rinsing or flushing cycles may be controlled by the controller 76.In this regard, both the cavity 118, the container 68 and the associated passages 116, 122, 120 and solenoid valve 104, 108 may be cleansed and sanitize periodically and automatically.","It will be appreciated that these automatic cycles will likely greatly increase the cleanliness, taste reliability and sanitation of such beverage dispensing systems as such procedures require considerable amount of time on the part of manual operators.","The use of controllable solenoid valves 104, 108 also prevents dispensing of beverage during a brew cycle.","For example, once a brew cycle is initiated the controller 76 can lock out the solenoid valves 104, 108.This will allow for the brewing cycle to complete and brewed beverage to mix within the container 68 to insure consistent flavor.","The beverage dispensing modules 32 can be constructed in any other suitable manner to dispense brewed beverage in any suitable manner in accordance with various embodiments disclosed.","If desired, for example, the beverage containers 68 of the beverage dispensing modules 32 can instead be in the form of carafes (not shown) or have any other suitable construction.","In a preferred embodiment, during a brewing cycle, heated water 134 is dispensed from the housing 34 through the water dispensing system.","Heated water 134 is used to infuse a beverage brewing substance 136 retained in the brewing assembly 64.An opening 138 is provided in the bottom of the funnel 66 to allow a brewed beverage 140 to flow therefrom.","As shown, for example, in previous figures, a brewed beverage container 68 is provided to collect the beverage 140.The beverage container 68 may be of any suitable form.","With reference to FIG.","8, a liquid flavor dispensing system 130 may be employed with the modular beverage dispensing system disclosed.","The liquid flavor dispensing system 130 can have any suitable configuration and can be used to supply flavoring 132 to all or less than all of the beverage dispensing modules 32.The liquid dispensing system 130 may also be used to supply a different flavoring 132 to each of the different beverage dispensing modules 32.Additionally, the flavorings 132 can be supplied to the beverage dispensing modules by a single liquid flavor dispensing system 130 or several liquid flavor dispensing systems.","Further, the flavoring 132 may in the form of the typical flavorings associated with brewed coffee or instead be in any other suitable form such as, for example, honey, peach, lemon, herbal flavors, or any other suitable flavoring which might be desired by a beverage consumer for any number of different beverages.","In a preferred embodiment, a small quantity of flavoring 132 is introduced into the water which is dispensed to the beverage brewing substance 136.The beverage brewing substance may be in the form of decaff or regular ground coffee as well as tea substances or other substances which might be used in an infusion beverage brewing process.","The objective in using the liquid flavoring material 132 is to introduce a small quantity of concentrated material or substance 132 during the brewing process so that the flavor becomes fully mixed with and saturated in the heated water 134.Introduction of the flavor 132 early in the brewing process will help assure that the flavor fully mixes with the water, is fully flushed through the beverage brewing substance to maximize the value of the flavor and is used to produce an aroma which might entice purchasers of the beverage as part of the beverage purchasing experience.","The flavor could also be dispensed into the beverage brewing substance and subsequently mixed with the water and the resultant brew during the brewing process.","A benefit of adding flavors at the time of brewing and/or mixing soluble beverages, over using pre-flavor beverage substances, is the increased effectiveness of the flavorings.","The increased effectiveness of the flavorings reduces the amount of flavoring required to produce a desired taste profile.","Under these circumstances, the flavorings are mixed with the beverage substance under optimal conditions to maximize the flavoring benefits of the flavorings.","For example, the time prior to mixing the flavoring with the brewing substance is nominal thereby virtually eliminating any degradation of the flavoring which might occur if the flavoring was exposed to air.","A liquid flavor dispensing system 130 is illustrated diagrammatically in FIG.","8.With this embodiment, the liquid flavor dispensing system dispenses a different liquid flavoring 132 in a controlled manner to several of the brewing assemblies of the beverage dispensing modules 32.The flavor can be dispensed to a water or liquid distribution system which controllably dispenses water to a beverage making substance, the beverage making substance or both.","With reference to FIGS.","8-10, the illustrated liquid flavor dispensing system 130 desirably includes a plurality of packages or containers 144a-144h, each of which retains a respective liquid flavor 132 therein.","The containers 144 desirably are in controlled communication with respective beverage dispensing modules 32a-32d in any suitable manner.","For example, a section of tubing or other passage 146 may connect each container 144 to a respective check valve 148.The check valve 148 is part of a respective solenoid or other form of pump device 150 coupled to the controller 76 via a respective line 152.A respective small diameter tube 154 extends from the solenoid pump 150 to a respective second check valve or flavor injection port 156 at a respective terminal end 158 of the tube 154.A respective sensor 160 can be coupled to the controller via a respective line 162 to monitor and detect a low level of the respective flavoring 132 in the respective container 144.As will be described in greater detail below with regard to sensing and monitoring, the status of the flavorings 132 can be reported by the controller 176.The check valves 148 and 156 at each end of each of the tubes 154 provide a sealed line to which the container 144 can be attached.","Also, either alternatively or in addition to the use of the sensor 160, the flavor usage can be inferred based on information collected on a characteristic of the pump such as operation or usage for example, pump actuation.","For example, a sensor may be associated with the pump and run time of the pump 150 could be monitored and a calculation made to infer the amount of flavorings remaining in a corresponding flavorings container 144.The system would provide a signal to the operator, such as “check-status” when the level of flavorings approaches, attains or drops below a predetermined level, for example 20% remaining.","Such advance warning would give the operator the opportunity to change the flavorings container 144 before it runs out.","The advance warning also helps assure that a reasonable margin of error in the inferred calculation is considered and accommodated for.","In use, when a brewing cycle is initiated, the solenoid pump 150 can be activated simultaneous with the initiation of the brewing cycle of one of the beverage dispensing modules or shortly thereafter.","A quantity, for example 9 ml for use in brewing ½ gallon of coffee, of liquid flavoring 132 is pumped from the container 144 through the tubing 154.The check valve or flavor injection port 156 is positioned in close proximity to a sprayhead 170 of the dispensing assembly 92.Dispensing of the flavor 132 into the water will allow the flavor to mix with the heated water for brewing substance during the saturation and extraction process.","Generally, it is desirable to use the flavoring at the start of the cycle which will result in completely integrating the flavor during the brewing cycle.","The process may be repeated with other beverage dispensing modules 32, 32a and, if desired, with other flavors.","The flavor injection ports 156 may, for example, include a check valve of a type as produced by Smart Products.","This is a positive pressure inverted check valve which requires positive pressure through the tube 154 to open the valve.","Positioning of the injection port 156 in close proximity to the sprayhead 170 results in the benefit of clearing and rinsing the surface as a result of the water vapor and heat during a brewing cycle.","Periodic manual cleaning of the sprayhead 170 and injection port 156 helps maintain a clean system.","Desirably, because the injection port 156 is an inverted pressure check valve, physical contact with the tip of the injection port 156 through which the liquid 132 is dispensed does not result in dispensing of liquid 132.Rather, physical contact with the end of the injection port 156 causes the valve to close as opposed to open.","The system also prevents the entry of atmosphere into the container 144 or line 154 thereby preventing contamination or degrading of the flavor or quality of the flavorings.","This system provides a generally closed system which prevents the flavorings contact with the atmosphere and prevents contamination providing a generally aseptic environment.","With reference to FIG.","11, the liquid flavor dispensing system 130 as described with regard to FIGS.","8-10 may be used with a soluble beverage system 176.In FIG.","11, the liquid dispensing system dispenses liquid flavor into a hot water path 174 of a soluble or powdered beverage dispensing system 176.The devices used to dispense the liquid flavor are the same as those describe above with regard to FIG.","8.As such, only the terminal ends 158a, 158b and 158c of multiple lines 154a, 154b, and 154c are shown.","Similarly, the multiple injection ports 156a, 156b, and 156c are shown.","The soluble beverage dispensing system 176 retains a quantity of powdered beverage mix (not shown) in a beverage dispensing container 178.The dispensing of soluble beverage material in powder form is generally known in the art.","The container 178 includes a dispensing auger (not shown) which controllably dispenses powder by operation of a dispensing drive or motor 180 controlled by the controller 76a.","Powdered beverage material is dispensed through the passage 182 into a mixing chamber 184.The hot water line 174 communicates with the mixing chamber 184 to combine with the powder.","The combined powder and hot water flow into an agitating chamber 186 there below.","A whipper motor 188 and agitating blade 190 further blend the powder and water combination for dispensing a beverage 192 through a nozzle 194.The flavor dispensing system 130 (see, FIG.","8) controllably dispenses liquid flavor 132 from respective containers 144 using corresponding pumps via the lines 154a, 154b, and 154c.","Either individual flavors or combinations of flavors may be dispensed into the hot water line.","Such dispensing may be programmed in the controller 76a.","Positioning of the injection ports 156a, 156b, 156c in the heated water path helps to assure that the liquid flavors are rinsed from the injection ports during and at the conclusion of the heated water dispensing cycle during a beverage dispensing cycle.","It should be noted that generally these powdered beverage dispensing systems have a momentary hot water overrun at the end of the cycle during which powdered beverages dispense from the container 178.The slight overrun of heated water helps to assure that the mixing chamber 184, whipping chamber 186, and nozzle 194 are rinsed clean at the end of the cycle.","The injection ports 156a, 156b and 156c have been positioned in the hot water line to provide the rinsing characteristics and benefits of the overrun of hot water.","Furthermore, it is commonly known to have at least one daily rinse cycle in which heated water is dispensed through the assembly 176 to maintain cleanliness and sanitation of the system.","During a brew cycle as shown in FIG.","8, liquid flavor is generally preferably dispensed at the initiation of the brew cycle.","As noted, this helps to ensure complete mixing and optimal extraction of the flavor from the beverage brewing substance 136.A predetermined volume of liquid flavor 132 can be dispensed into the brewing assembly 30 at the initiation of the brew cycle.","In contrast, the soluble system 176 as shown in FIG.","11 may have a variable quantity of beverage 192 dispensed therefrom.","It is anticipated that multiple beverage sizes may be dispensed from the soluble system 176.As a result, the liquid flavoring 132 must be dispensed at a rate consistent with the concentration flavor profile or recipe, and powder also dispensed by the system.","For example, if a consumer wishes to obtain a 12 ounce beverage the proper amount of liquid flavoring must be dispensed for a 12 ounce beverage.","However, the consumer may be allowed to control whether they wish to have a full 12 ounces of beverage or if they decide to short the beverage resulting in, perhaps, only 10 ounces being dispensed.","As such, the pump associated with the powdered beverage system 176 must be calibrated to dispense small quantities of liquid flavoring 132 on a per unit time basis to assure a desirable and consistent mixing of flavor with the powder.","This will help assure that the desired flavor is achieved regardless of the quantity of beverage dispensed.","The solenoid pump 150 used with the liquid flavor dispensing system 130 is a precision controlled metering pump.","It is anticipated that other forms of the metering pump may be used with this system.","However, in the present embodiment, the metering pump is controllably operated to dispense very small quantities of the liquid flavor 132.The frequency of the plunger rate, the stroke of the plunger as well as the duration of plunger operation the solenoid pump 150 can be controlled by the controller 76, 76a.","As a result, very small quantities of highly concentrated liquid flavor can be dispensed into the beverages 140, 192 during the corresponding beverage preparation process.","As such, precise quantities can be added to provide flexibility and tailoring of the resultant beverages.","The system 130 can be calibrated using volumetric guidelines associated with various beverage recipes and flavor materials to match particular flavor preferences, for example for various demographics.","As such, a franchise operation can custom blend and configure the flavor of beverages dispensed by the beverage dispensing system to meet and satisfy regional preferences.","Additionally, if preferences change over time the system can be modified and calibrated to match those preferences.","This adds further flexibility to the beverage system disclosed.","With reference to FIGS.","14-18, a powdered or soluble beverage dispensing system 199 is shown.","While a soluable beverage system 199 has been shown and described as a powder dispensing system, it is envisioned that such a soluable system also includes a liquid concentrate system which dispenses a quality of liquid coffee concentrate for mixing with water or other dilution or reconsituting substance.","Reference to a soluable beverage system is intended to be broadly interpreted.","Such a beverage dispensing system has been generally and diagrammatically shown in FIG.","11.With further reference to FIG.","14, the container 178 is shown positioned inside a housing 200.The passage 182 coupled to the container 178 is shown positioned for dispensing into the mixing chamber 184 and communicating with the whipping chamber 186 and nozzle 194.A portion of the housing 200 in the form of a door 202 conceals the powdered beverage system 176 in the housing 200.Also, the liquid flavor dispensing system 130 as described hereinabove may be used with the powdered beverage or soluble beverage dispensing system 176 illustrated in FIGS.","14-18.One of the potential difficulties involved with a soluble beverage dispensing system is that the container 178 must be removed periodically for refilling with powdered or soluble beverage substance as well as regular maintenance and cleaning.","The container 178 may not be very heavy when it is empty because it is generally formed of a plastic material.","However, once filled, it may contain 5 to 10 pounds of powder material.","With the container 178 being positioned in an elevated location and containing a quantity of material, it may be difficult to lift the container into a desired position in the housing 200.The container 178 is positioned in an upper position in the housing 200 in order to benefit from gravity assistance when dispensing powdered beverage therefrom to the mixing chamber 84.Additionally, it is important to align the dispensing passage 182 with the housing 184 to help assure that when powdered beverage material is dispensed from the container 178 it flows into the proper path for mixing with water.","As such, it would be desirable to provide an apparatus and system for facilitating improved ease of removal and replacement of the powdered beverage container 178 relative to the housing 200.As shown in FIGS.","14-18, a container positioning assembly or container positioner 220 in the form of a slide-out support or shelf is provided.","The container positioning assembly 220 includes a pair of handles 222 which can be grasped (see FIG.","15) by a user to extract the container 178 attached to the assembly 220 for removing it from the housing 200.As shown in FIG.","15, the assembly 220 allows the container 178 to be pulled straight out of the housing 200 thereby allowing disengagement of the passage 182 relative to the mixing chamber 184.Movement of the assembly 220 also replaces the container 178 in a desired position to align the dispensing passage 182 with the mixing chamber 184.As can be seen in FIGS.","15-17, the container 178 has been removed from the housing 200 in order to refill the container 178 but does not have to be removed from the shelf 250 of the assembly 220.Support rails 226 are positioned on each side of container 178 and are engaged with side walls 228 of the housing 200 (FIG.","15).","Each of the guide rails 226 includes a pair of tracking slots 230, 232 which are engaged with corresponding upper 234 and lower 236 guide rollers.","A support roller 240 is positioned along an exterior edge 242 of the guide 226 to further help facilitate smooth and controlled movement of the assembly 220.Support rollers 240 and guide rollers 234, 236 are mounted on the sidewalls of the housing 200.The guides 226 are connected by a cross extending shelf 250 on which the container 178 is positioned.","The combination of the guide rails 226 and rollers help the assembly 220 slide outwardly and inwardly relative to the housing 200.FIG.","16 shows only the portion of the guide rails 226 extending past the edge of container 178.The guide rollers 240, 234, 236 associated with the guides 226 prevent tipping of the shelf 250 and associated container 178.When the assembly 220 is stowed in the housing 200 the shelf 250 rests on a corresponding surface 252 of the housing 200.As can be seen in FIGS.","15-17, the guides 226 include a generally horizontal portion 254 and a vertically oriented portion 256.The shelf 250 and container 178 are associated with the generally horizontal portion 254.The generally vertical portion 256 helps to allow forward movement as well as downward movement of the assembly 220 relative to the housing 200.The guide including the portion 254, 256 help facilitate movement of the container into and out of the housing 200 and forward and downward movement.","In the embodiment as shown, the assembly facilitates positioning of the container 178 approximately 6-8 inches downwardly from its stored position in the housing 200.This lowering of the container 178 is beneficial in that it allows for easy filling of the container.","Also, by fixing the container to the shelf 250 and the assembly 200 it prevents spilling or otherwise tipping of the container 178 during filling or other processes.","This can result in savings in terms of lost beverage material as well as cleanup costs and associated complications.","In the stored position, detents 260, 262 are provided on the guides 230, 232, respectively.","The detents 260, 262 allow the rollers 234, 236 respectively, to be engaged therewith to prevent casual disengagement of the assembly 220 from the housing 200.This helps to lock or retain the assembly 220 in a desired stored or stowed position in the housing 200.As the assembly 220 is moved outwardly from the housing 200, the track guides 234, 236 track along the corresponding channels 230, 232 generally horizontally along the horizontal portion 254.As such, the container 278 attached to the shelf 250 sides forwardly and outwardly from the housing.","At a position where the generally horizontal portion 254 transitions into the partially vertical portion 256 (elbow 270) the guides allow the assembly to slide downwardly and forwardly.","In the extended most position the guides 234, 236 engage corresponding extended detents 272, 274, respectively.","These detent positions 272, 274 help retain the assembly 220 in the outward or extended most position.","The assembly 220 also includes an assist system 280 which includes cables 282 and biasing members to springs 284 forming a cable-biasing assembly help facilitate removal of and replacement of the container 178 relative to the housing 200.With reference to FIG.","18, the assist system 280 includes a spring 284 attached at one 286 to a back portion of the housing and at a second end to a first pulley 288.The cable 282 is attached at a first end 290 to a corresponding portion of the housing and extends through the first pulley 288 and up and around a second pulley 292.The second end 294 of the cable 282 attaches to a corresponding portion of the guides 226 at attachment point 296.Attachment of the cable 282 to the guide 226 is shown in FIG.","16.The assist assembly 288 helps further facilitate ease of removal and replacement of the container 178 relative to the housing 200.For example, when the container 178 is removed from the housing on the assembly 220 the spring 284 stores energy and is stretched as the cables 282 are extended.","The cables 282 are extended as a result of the forward and downward movement of the guides 226.Extension of the springs 284 expends the stored energy and imparts a return force in the springs which helps reduce the force required to return the container 178 to the housing 200.This is particularly useful when an empty container is removed from the housing, filled with powdered material and then returned.","Returning the filled container 178 normally would require additional effort on the part of the operator and as such the return forces in the assist system 280 help make the return easier.","Turning now to FIGS.","19 and 20, the beverage system also includes a communications system or means 300 for communicating 302 between a corresponding beverage brewing, beverage making apparatus or beverage dispensing modules 32, 32a and a receiver or base station 304.Each of the modules 32, 32a includes a controller 76, 76a which is centralized for use with multiple modules or dedicated to an individual module.","The controller collects information about the module or modules.","For purposes of this present discussion, we will refer to the controller as being dedicated to multiple modules and receiving information from each individual module associated therewith.","The many modules associated with a single control also provides the added benefit of the modular assembly as described hereinabove.","It should also be noted that the details of the circuitry used to achieve the communication system are illustrated in the detailed schematics provided in FIGS.","21-28 and all corresponding sub-portions thereof.","For example, FIG.","21 is a general schematic of the overall system.","FIGS.","22-28 provide details about the general system shown in FIG.","21.FIG.","21 has been noted with cross reference to the other figures which provide details about the portions of the circuitry shown generally in FIG.","21.The controllers 76, 76a include the link 302 which connects the controller to the corresponding receiver or base station 304.In the present embodiment as diagrammatically illustrated in FIGS.","19 and 20, the controllers 76, 76a communicate with the receiver 304 by way of an RF link 302.Generally, the controllers are remote from the receiver and use the Link 302 and the associated means for linking to communicate the collected information about the beverage making apparatus.","Antennae 306, 308 are provided on the controllers 76, 76a and the corresponding receivers 304 respectively.","It should be noted that the communication link 302 may be in any one of a variety of forms such as hardwired physical point-to-point link, optical lines, light wave, ultrasonic, infrared or any other form of communication link between one or more devices.","The receiver 304 receives information from the controller or various components of the modules 32, 32a to identify information relating to the modules and operation of the modules.","The receiver 304 may be in the form of a monitoring unit positioned in an appropriate location relative to an operator or attendant who has responsibility for or otherwise maintains the modules 32, 32a.","The receiver 304 includes a means for displaying display 305 and can provide visual auditor or other information about numerous conditions.","The display 304 allows a form of the collected information about the beverage making apparatus to be displayed at the receiver.","The visual display may include, but is not limited to lights, text, symbolic images (i.e., dispenser “full” showing a colored or shaded dispenser and a dispenser “empty” showing an unshaded dispenser), and mechanical devices that tare operated or other visual displays.","An auditory display may include, but is not limited to speech information, alarms, tones or other signals that van be heard.","Tactile displays are also contemplated in the form of a vibrating surface, vibrating device worn by the operator or other means for letting the operator or other means for letting the operator know about he collected information.","For example, the following is a list of the messages or conditions which can be displayed: Receiver fault; Please wait attempting communication; Communication failure, coffee module #; Communication fault, soluble module #; Flavor [type] low, coffee module #; Flavor [type] low, soluble module #; Hopper low, soluble module #; Hopper empty, soluble module #; Coffee empty, coffee module #; Coffee low, coffee module #; Freshness expired, coffee module #; Dumping in minutes, coffee unit #; Server removed, coffee module #; Refill too long; Temperature probe open; Temperature probe short; Heating too long; Over flow safety; Coffee stations all OK; and Soluble stations all OK. For example, in an convenience store setting, multiple modules 32, 32a may be generally remotely positioned in one portion of the store, for example, a rearward location of the store, if desired.","The operator of the beverage system may also be the same person responsible for stocking of the equipment, and register activities.","As such, one person is required to operate and maintain many components of the convenience store facility.","The receiver 304 may be positioned at the cash register so that while the attendant is operating the cash register he may also be notified of matters that require his attention at the modules 32, 32a.","For example, if a module 32a indicates that a container 68 is empty the controller 76 will communicate with the receiver 304 to indicate this condition.","The receiver may also be configured with audible alarms and visual displays to provide additional information.","For example, a display may identify which module and the specific condition associated with the module which has provided a signal to the receiver 304.The operator can then make a decision how to use that information relative to his responsibilities at the cash register.","If an emergency requires immediate attention he can secure the cash register and address the emergency while attending to the module 32.Alternatively, if it is a slow period in the store and a module is indicating that a container 68 is empty he can put off attending to this condition until he completes one or more transactions in queue.","The signal communication between the controller 76 and the receiver 304 may be a single path signal or in the form of a multiple path verification signal.","For example, as shown in FIGS.","19 and 20 a first signal 310 is transmitted from the controller 76 to the receiver 304.This initial communication may report the status of a condition at the module 32.The receiver 304 can then return a copy of the information 312 to the controller 76.If the controller confirms the information received (312) it can then send a return confirmation 314 that the information is correct.","This is a useful communication link in the present situation due to the presence of electrical noise and other interference within other beverage system applications.","For example, each of the components associated with the modules 32, 32a may provide some degree of electrical or other noise.","Additionally, other systems within the beverage system setting may be producing RF or other signals which could create interference.","As such, the present embodiment providing the verification communication links 310-314 helps facilitate and assure accurate and timely communication.","In the event of excessive interference such as from patrons of the store using telephone or RF communication devices, the controller 76 can continue to periodically send a message 310 to the receiver 304 until the verification signal 312 is received.","While preferred embodiments are disclosed, illustrated and described, it is envisioned that those skilled in the art may devise various modifications and equivalents without departing from the spirit and scope of the disclosure as recited in the following claims."]],"string":"[\n [\n \"Liquid flavor beverage system\",\n \"A beverage apparatus (30) having a plurality of dispensing modules (32) and a housing (34) that defines a chamber (36) containing water or liquide suitable for use in brewing.\"\n ],\n [\n \"1.A liquid flavor dispensing system for use with a beverage making apparatus, the liquid flavor dispensing system comprising: a container for retaining a liquid flavoring; a passage connected to the container and extending from the container to a portion of a beverage making apparatus; a pump positioned along the passage for moving liquid flavor from the container to a portion of the beverage making apparatus; and means for controlling flow from the passage, the flow control means communicating with the passage for controlling flow therefrom.\",\n \"2.The liquid flavor dispensing system as in claim 1, further comprising means for monitoring a quantity of liquid flavor in the container.\",\n \"3.The liquid flavor dispensing system as in claim 2, wherein the monitoring means includes a sensor associated with the container for detecting the quantity of liquid flavor in the container.\",\n \"4.The liquid flavor dispensing system as in claim 1, wherein the monitoring means includes a sensor associated with the pump for detecting a characteristic of the pump for inferring the quantity of liquid flavor in the container.\",\n \"5.A beverage making apparatus comprising: a liquid distribution system; means for combining a beverage making substance with a liquid communicating with the liquid distribution system; a liquid flavor dispensing system for dispensing liquid flavor to at least one of the liquid distribution system and the combining means.\",\n \"6.The beverage making apparatus as in claim 5, further comprising the combining means including a beverage brewing portion for combining liquid from the liquid distribution system with a beverage brewing substance retained in the beverage brewing portion.\",\n \"7.The beverage making apparatus of claim 5, further comprising the combining means including a powdered beverage mixing portion for combining liquid from the liquid distribution system with a powdered beverage substance.\",\n \"8.The beverage making apparatus of claim 5, further comprising the combining means including a liquid beverage concentrate mixing portion for combining liquid from the liquid distribution system with a liquid beverage concentrate.\",\n \"9.The beverage making apparatus of claim 5, further comprising: the liquid distribution system having an outlet proximate to the combining means; and the liquid flavor dispensing system dispensing liquid flavor to the outlet of the liquid distribution system prior to combining with a beverage making substance.\",\n \"10.The beverage making apparatus of claim 5, further comprising: the liquid distribution system having a dispensing line for delivering liquid to the combining means; and The liquid flavor dispensing system dispensing liquid flavor to the dispensing line for mixing flavor with the liquid prior to mixing the liquid with the beverage making substance.\",\n \"11.The beverage making apparatus of claim 5, further comprising: the liquid flavor dispensing system dispensing liquid flavor to the combining means for dispensing flavor into the beverage making substance.\",\n \"12.A method of combining liquid flavor for use with a beverage making apparatus, the method comprising the steps of: providing a liquid flavor; providing a passage for dispensing the liquid flavor to a beverage making apparatus; dispensing a quantity of liquid flavor through the passage; and combining the dispensed liquid flavor with a beverage for modifying the flavor of the beverage.\",\n \"13.The method of combining a liquid flavor for use with a beverage making apparatus of claim 12, further comprising the steps of: providing a liquid distribution system in combination with the beverage making apparatus; and dispensing liquid flavor to a location proximate to the dispensing of liquid from the liquid distribution system for combining the liquid flavor with the liquid prior to combining with the beverage making substance.\",\n \"14.The method of combining a liquid flavor for use with a beverage making apparatus of claim 12, further comprising the steps of: providing a liquid distribution system having a dispensing line for delivering liquid to combine with a beverage making substance; and dispensing the liquid flavor to the dispensing line for mixing the flavor with the liquid prior to mixing the liquid with the beverage making substance.\",\n \"15.The method of combining a liquid flavor for use with a beverage making apparatus of claim 12, further comprising the step of: dispensing liquid flavor to the beverage making substance.\"\n ],\n [\n \"<SOH> BACKGROUND <EOH>This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.\",\n \"Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.\",\n \"In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.\",\n \"Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.\",\n \"As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.\",\n \"Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.\",\n \"Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.\",\n \"Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.\",\n \"These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.\",\n \"To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.\",\n \"In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process.\"\n ],\n [\n \"<SOH> SUMMARY <EOH>The present disclosure relates to apparatus for making and dispensing beverages.\",\n \"The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.\",\n \"Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.\",\n \"The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.\",\n \"In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.\",\n \"In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.\",\n \"The apparatus may also include a plurality of conduits for supplying liquid to the assembly.\",\n \"The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.\",\n \"Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.\",\n \"The housing may be positioned behind the beverage dispensing modules.\",\n \"In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.\",\n \"With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.\",\n \"The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.\",\n \"A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.\",\n \"The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.\",\n \"The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.\",\n \"In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.\",\n \"In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.\",\n \"Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived.\"\n ],\n [\n \"BACKGROUND This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.\",\n \"Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.\",\n \"In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.\",\n \"Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.\",\n \"As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.\",\n \"Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.\",\n \"Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.\",\n \"Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.\",\n \"These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.\",\n \"To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.\",\n \"In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process.\",\n \"SUMMARY The present disclosure relates to apparatus for making and dispensing beverages.\",\n \"The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.\",\n \"Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.\",\n \"The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.\",\n \"In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.\",\n \"In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.\",\n \"The apparatus may also include a plurality of conduits for supplying liquid to the assembly.\",\n \"The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.\",\n \"Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.\",\n \"The housing may be positioned behind the beverage dispensing modules.\",\n \"In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.\",\n \"With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.\",\n \"The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.\",\n \"A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.\",\n \"The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.\",\n \"The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.\",\n \"In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.\",\n \"In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.\",\n \"Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS The detailed description particularly refers to the accompanying figures in which: FIG.\",\n \"1 is a partial fragmentary side elevational view of a modular beverage dispensing apparatus; FIG.\",\n \"2 is a top plan view of the modular beverage dispensing apparatus of FIG.\",\n \"1; FIG.\",\n \"3 is a side elevational view of the modular beverage dispensing apparatus of FIGS.\",\n \"1 and 2; FIG.\",\n \"4 is a schematic illustration of the modular beverage dispensing apparatus; FIG.\",\n \"5 is a schematic illustration of one module of the modular beverage dispensing apparatus; FIG.\",\n \"6 is a diagrammatic illustration of a dispensing and disposal valve configuration; FIG.\",\n \"7 is a partial fragmentary top plan view taken along 7-7 in FIG.\",\n \"6; FIG.\",\n \"8 is a diagrammatic illustration of a liquid flavor distributing system; FIGS.\",\n \"9 and 10 are diagrammatic illustrations of the arrangement of liquid flavor distributing systems; FIG.\",\n \"11 is a cross sectional view, schematic in nature, of a portion of a powdered beverage, soluble beverage dispensing system including connections to dispense liquid flavor to the soluble beverage production stream; FIGS.\",\n \"12 and 13 are diagrammatic illustrations of the liquid flavor distributing system coupled to the soluble beverage dispensing system of FIG.\",\n \"11; FIG.\",\n \"14 is a perspective view of a soluble beverage dispenser having a slide-out assembly; FIG.\",\n \"15 is an enlarged perspective view of the soluble beverage dispenser of FIG.\",\n \"14 showing a user removing a container from the soluble beverage dispensing device using the slide-out assembly; FIGS.\",\n \"16 and 17 are enlarged views of portions of the slide-out assembly of the soluble beverage dispenser of FIG.\",\n \"14; FIG.\",\n \"18 is a perspective view of a cable and spring leverage system which assists movement of the slide-out assembly relative to the beverage dispensing system; FIGS.\",\n \"19 and 20 are schematic illustrations of a communication link between a base station and a beverage dispensing device; and FIGS.\",\n \"21-28 are schematic illustrations of the circuitry associated with the communication system of FIGS.\",\n \"19 and 20 AND INCLUDE FIG.\",\n \"22A, FIGS.\",\n \"22-22D, FIGS.\",\n \"23-23D, FIG.\",\n \"24, FIGS.\",\n \"24A-24B, FIGS.\",\n \"25A-25B, FIGS.\",\n \"26A-26, FIG.\",\n \"27, FIGS.\",\n \"27A-27C, FIG.\",\n \"28 and FIGS.\",\n \"28A-28E.\",\n \"DESCRIPTION OF THE DRAWINGS While the present disclosure may be susceptible to embodiment in different forms, there is shown in the drawings, and herein will be described in detail, embodiments with the understanding that the present description is to be considered an exemplification of the principles of the disclosure and is not intended to limit the disclosure to the details of construction and the arrangements of components set forth in the following description or illustrated in the drawings.\",\n \"A beverage dispensing apparatus 30 illustrated generally in FIGS.\",\n \"1-4 as comprising a plurality of beverage dispensing modules 32 and a housing 34 that defines a chamber 36 containing water or other liquid suitable for use in brewing.\",\n \"The housing 34 desirably acts as a central reservoir for a constant supply of heated water to be used by the beverage dispensing modules 32 to brew several different beverages.\",\n \"The beverage dispensing apparatus 30 may have any suitable construction and configuration.\",\n \"The beverage apparatus 30 may include any suitable number of beverage dispensing modules 32 oriented in any suitable manner, depending on various circumstances, such as, for example, customer demand, customer usage, available store space, and desired aesthetics.\",\n \"Similarly, the housing 34 may have any suitable construction and configuration that desirably is consistent with customer demand, customer usage, available store space and desired aesthetics.\",\n \"A plurality of conduits in the form of water lines 38 provide flow communication between the housing 34 and the beverage dispensing modules 32 to supply water for brewing.\",\n \"In a preferred embodiment, the housing 34 is positioned behind the beverage dispensing modules 32 and centrally located relative to the beverage dispensing modules 32 to minimize the distance between the housing and each of the beverage dispensing modules, thereby increasing operational efficiency and minimizing space.\",\n \"Terms including beverage and beverage making as used herein are intended to be broadly defined as including but not limited to the brewing of coffee, tea and any other brewed beverage, including a final beverage or food product as well as producing an intermediate product to be combined with a final beverage or food product.\",\n \"This broad interpretation is also intended to include, but is not limited to any process of infusing, steeping, reconstituting, diluting, dissolving, saturating or passing a liquid through or otherwise mixing or combining a beverage substance with a liquid such as water without a limitation to the temperature of such liquid unless specified.\",\n \"This broad interpretation is also intended to include, but is not limited to beverage substances such as ground coffee, tea, liquid beverage concentrate, powdered beverage concentrate, freeze dried coffee or other beverage concentrates, to obtain a desired beverage or other food.\",\n \"In the illustrated embodiment, for example, the beverage dispensing apparatus 30 comprises ten beverage dispensing modules 32 oriented in a generally U-shaped manner that substantially surrounds the housing 34.As illustrated, four of the beverage dispensing modules 32 face the same direction, four other beverage dispensing modules 32 face an opposite direction, and two of the beverage dispensing modules 32 are positioned in a space-efficient manner consistent with a desired orientation.\",\n \"A simplified form of the beverage apparatus 30 is shown in FIG.\",\n \"4 which uses simplified block diagram illustrations to show multiple beverage dispensing modules 32 which are configured to provide a beverage brewing system.\",\n \"The beverage apparatus 30 shown in FIG.\",\n \"4 is very similar to a soluble beverage apparatus 30a also shown in FIG.\",\n \"4.The beverage apparatus 30, as will be described in greater detail herein below, relates to a beverage brewing or producing system whereas the soluble beverage apparatus 30a, as will be described in greater detail herein below, relates to a soluble or powdered beverage dispensing system.\",\n \"Both beverage apparatuses 30, 30a employ a common heated water reservoir contained in housing 34 to which the individual modules 32, 32a are connected.\",\n \"As shown in FIGS.\",\n \"2-4, the housing 34 desirably is centrally located to serve multiple beverage modules 32, 32a.\",\n \"The benefit of a common housing 34 functioning as a common reservoir to supply water, instead of individual tanks for each module 32, 32a, is that it substantially reduces the cost and greatly increases the efficiency of the system.\",\n \"Also it reduces the redundancy of multiple heated water reservoirs, reduces the wattage required by the system and increases flexibility in the number of modules 32, 32a which might be included.\",\n \"The particular configuration with the housing 34 positioned in close proximity to or between several modules 32, 32a is beneficial in that it also minimizes the length of water lines 38 connecting the housing 34 to the modules 32, 32a.\",\n \"Additionally, a larger body of water in the housing 34 tends to help maintain the temperature and reduce temperature loss over time as well as maintaining a larger volume of brew water at a desired temperature.\",\n \"While the housing 34 retained within the beverage apparatus 30 is insulated to reduce heat loss and reduce heat load on the ambient environment, heat which may escape from the housing 34 will generally pass to the modules 32, thereby increasing the efficiency of maintaining the temperature of the beverages retained in the modules 32.Insulation desirably is provided to reduce beat loss to the ambient environment.\",\n \"This will also help reduce the exertion on the ambient air-conditioning system thereby further increasing efficiency of the present beverage apparatus 30, 30a.\",\n \"With reference to FIGS.\",\n \"1-4, each module 32, 32a produces a separate brewed beverage.\",\n \"The centralization of a housing 34 helps reduce the individual cost associated with the module 32,32a and improves the ease of manufacturing of the modules.\",\n \"Also, there is increased commonality of parts associated with this type of system.\",\n \"The modular configuration of the beverage apparatus 30 using the modules 32, 32a increases the potential variability of the physical size and brewer configuration of the system.\",\n \"This is beneficial to accommodate the needs of a particular location.\",\n \"In other words, only a desired or required number of modules 32, 32a will be needed at any given time.\",\n \"If a location has an increase in demand for brewed or soluble beverages additional modules 32, 32a may be added as necessary for the particular change in demand.\",\n \"Additionally, with regard to repair and maintenance, the beverage apparatus 30, 30a can be designed such that individual modules 32, 32a can be removed for repair and maintenance as necessary with an identical module being inserted in the missing modules.\",\n \"In a preferred embodiment, (FIG.\",\n \"5) the housing 34 includes a heating element 52, and one or more system sensors such as a level sensor 86 and a temperature sensor 84.Such components tend to be generally readily available and easily replaced and as such further enhance the reliability of the overall beverage apparatus 30, 30a.\",\n \"With reference to FIG.\",\n \"3, the line 38 connecting each housing 34 to a brewing portion 40 of the module 32 is shown.\",\n \"Water 42 retained in the housing 34 can be readily dispensed.\",\n \"A water inlet line 44 is provided to introduce unheated water 50 to the housing 34.The inlet line 44 runs from an upper portion 46 of the housing 34 and directs the inward water downwardly towards a bottom portion 48 of the housing 34.Introducing the unheated water 50 towards the bottom of the reservoir introduces the unheated water 50 in close relation to a heated element 52 thereby accelerating the heating of the water.\",\n \"Additionally, the introduction of unheated water 50 in the bottom portion 48 tends to force heated water in the upper portion 46 upwardly.\",\n \"As such, the water 42 in the upper portion 46 tends to be the hottest water in the housing 34.Introducing unheated water 50 from the upper portion has the benefit of providing an entrance into the reservoir through a reservoir wall 56 with little or no pressure on the entry therethrough.\",\n \"This may result in an increase in the reliability of the entry therethrough.\",\n \"Additionally, a reinforcing or strengthening structure 58 is provided extending between opposed walls 60, 62 of the housing 34.This helps prevent bowing or stress on the walls 60, 62 which might otherwise result from the force of the water on the walls.\",\n \"Multiple reinforcing structures 58 may be provided throughout the housing 34 as necessary to provide support for the size and dimension of the housing required for a particular system.\",\n \"FIG.\",\n \"5, for example, shows a diagrammatic illustration of an embodiment of the apparatus 30 shown in FIGS.\",\n \"1-4.The beverage dispensing modules 32 can have any suitable construction desirably adapted to produce beverages, store beverages for ready consumption, and facilitate ready dispensing of the beverages.\",\n \"Each module 32 includes a brewing assembly 30 that may have any suitable construction that can be used to produce beverage such as brewing coffee or other beverages.\",\n \"The illustrated brewing assembly 30, for example, includes a brewing substance retainer or assembly 64 including a funnel 66.Such a retainer 64 may be well known in the art and may include a filter (not shown) retained in the funnel 66 for holding a charge of beverage substance.\",\n \"The funnel 66 may be positioned above a reservoir or container portion 68 of the dispensing module 32 which receives brewed beverage from the funnel.\",\n \"A dispensing assembly 70 in the form of a controllable valve may be associated with the container 68 for dispensing beverage from the module 32.With reference to FIG.\",\n \"5 a simplified review of the beverage brewing process is described.\",\n \"A brew actuator switch 72 desirably is associated with each of the modules 32.To initiate a brew cycle for one of the modules 32 a respective brew activation switch 72 is operated.\",\n \"Operation of the switch 72 sends a signal over line 74 to controller 76.Controller 76 operates an inlet valve 78 over line 80 to introduce water to the housing 34.Also, heating element 52 is coupled to the controller 76 over line 82.The temperature sensor 84 and level sensor 86 are coupled to the controller over lines 88, 90 respectively.\",\n \"If the controller 76 detects proper level and temperature it will initiate a brew cycle thereby displacing water from the housing 34 through a sprayhead assembly 92.The sprayhead assembly 92 communicates with the substance retainer 64 and dispenses water into the funnel 66 for producing a beverage therein.\",\n \"The illustrated module 32 includes a level sensor 94 associated with the container 68 which is coupled to the control over line 96.The level sensor can detect a change in level in beverage retained in the container 68 which may be used to activate a brew cycle.\",\n \"Also, the level detector 94 may sense the level in the beverage container 68 thereby indicating if the container is full or above the desired level to prevent initiation of a brew cycle.\",\n \"This will prevent over flow of the beverage server 68.The beverage dispensing modules 32 can be constructed to dispense beverage in any suitable manner.\",\n \"In a preferred embodiment, for example, a dispenser or faucet switch 98 may be associated with the dispensing assembly 70 and coupled with a dispensing handle or control 100.Activation of the control 100 will activate the switch 98 over line 102 connected to the controller 76 thereby indicating that the controller 76 should operate a solenoid valve 104 (FIGS.\",\n \"6 and 7), via line 106 to dispense beverage therefrom.\",\n \"As will be described in greater detail below with regard to FIGS.\",\n \"6 and 7, a disposal or dump valve 108 is also coupled with the container 68 to facilitate controllable disposal, via line 110 coupled to the controller 76.With reference to FIGS.\",\n \"6 and 7, the beverage container 68 may further include a manifold 114.The illustrated manifold 114 includes a primary passage 116 which communicates with an interior cavity or reservoir 118 of the container 68.Beverage flows from the cavity 118 through the primary passage 116.The solenoid valves 104, 108 communicate with the passage 116.A dispensing passage 120 and a disposal passage 122 communicate with the primary passage 116.Solenoid valve 104 and 108 desirably are normally closed and require specific activation in order to open the valves.\",\n \"During a dispense cycle the switch or control 100, for example a controllable faucet as shown in FIG.\",\n \"5, is activated thereby activating solenoid valve 104.When the dispense solenoid valve 104 is opened beverage 124 will flow from the cavity through the solenoid valve 104.Under this condition disposal valve 108 is closed.\",\n \"When beverage must be disposed of, the disposal solenoid valve 108 is opened while maintaining the dispense solenoid valve 104 in a closed position.\",\n \"This will allow the disposal of beverage through a separate dispensing line.\",\n \"Disposal of beverage may occur as a result of a time lapsed period whereby after a specified period of time beverage should be disposed of.\",\n \"For example, if a beverage only maintains a desired flavor profile or other characteristics for a period of time, for example 2 hours, at the end of two hours after the beverage brewing cycle is initiated, the beverage may be disposed of through the disposal valve 108.Such disposal may also be used at the end of a service cycle such that at the end of the day all beverages may be disposed of at a predetermined time.\",\n \"The disposal valve may also be used in a manner to flush and cleanse the beverage brewing system.\",\n \"In this regard, at a predetermined period of time, the controller 76 may operate the disposal valve 108 to dispose of any beverage retained in the container 68.The disposal valve 108 and dispense valve 104 may be closed.\",\n \"The controller 76 may then dispense a quantity of water into the cavity 118 thereby flushing and sanitizing the reservoir.\",\n \"After holding the heated water for a predetermined period of time the water may be disposed through the disposal valve 108.Multiple rinsing or flushing cycles may be controlled by the controller 76.In this regard, both the cavity 118, the container 68 and the associated passages 116, 122, 120 and solenoid valve 104, 108 may be cleansed and sanitize periodically and automatically.\",\n \"It will be appreciated that these automatic cycles will likely greatly increase the cleanliness, taste reliability and sanitation of such beverage dispensing systems as such procedures require considerable amount of time on the part of manual operators.\",\n \"The use of controllable solenoid valves 104, 108 also prevents dispensing of beverage during a brew cycle.\",\n \"For example, once a brew cycle is initiated the controller 76 can lock out the solenoid valves 104, 108.This will allow for the brewing cycle to complete and brewed beverage to mix within the container 68 to insure consistent flavor.\",\n \"The beverage dispensing modules 32 can be constructed in any other suitable manner to dispense brewed beverage in any suitable manner in accordance with various embodiments disclosed.\",\n \"If desired, for example, the beverage containers 68 of the beverage dispensing modules 32 can instead be in the form of carafes (not shown) or have any other suitable construction.\",\n \"In a preferred embodiment, during a brewing cycle, heated water 134 is dispensed from the housing 34 through the water dispensing system.\",\n \"Heated water 134 is used to infuse a beverage brewing substance 136 retained in the brewing assembly 64.An opening 138 is provided in the bottom of the funnel 66 to allow a brewed beverage 140 to flow therefrom.\",\n \"As shown, for example, in previous figures, a brewed beverage container 68 is provided to collect the beverage 140.The beverage container 68 may be of any suitable form.\",\n \"With reference to FIG.\",\n \"8, a liquid flavor dispensing system 130 may be employed with the modular beverage dispensing system disclosed.\",\n \"The liquid flavor dispensing system 130 can have any suitable configuration and can be used to supply flavoring 132 to all or less than all of the beverage dispensing modules 32.The liquid dispensing system 130 may also be used to supply a different flavoring 132 to each of the different beverage dispensing modules 32.Additionally, the flavorings 132 can be supplied to the beverage dispensing modules by a single liquid flavor dispensing system 130 or several liquid flavor dispensing systems.\",\n \"Further, the flavoring 132 may in the form of the typical flavorings associated with brewed coffee or instead be in any other suitable form such as, for example, honey, peach, lemon, herbal flavors, or any other suitable flavoring which might be desired by a beverage consumer for any number of different beverages.\",\n \"In a preferred embodiment, a small quantity of flavoring 132 is introduced into the water which is dispensed to the beverage brewing substance 136.The beverage brewing substance may be in the form of decaff or regular ground coffee as well as tea substances or other substances which might be used in an infusion beverage brewing process.\",\n \"The objective in using the liquid flavoring material 132 is to introduce a small quantity of concentrated material or substance 132 during the brewing process so that the flavor becomes fully mixed with and saturated in the heated water 134.Introduction of the flavor 132 early in the brewing process will help assure that the flavor fully mixes with the water, is fully flushed through the beverage brewing substance to maximize the value of the flavor and is used to produce an aroma which might entice purchasers of the beverage as part of the beverage purchasing experience.\",\n \"The flavor could also be dispensed into the beverage brewing substance and subsequently mixed with the water and the resultant brew during the brewing process.\",\n \"A benefit of adding flavors at the time of brewing and/or mixing soluble beverages, over using pre-flavor beverage substances, is the increased effectiveness of the flavorings.\",\n \"The increased effectiveness of the flavorings reduces the amount of flavoring required to produce a desired taste profile.\",\n \"Under these circumstances, the flavorings are mixed with the beverage substance under optimal conditions to maximize the flavoring benefits of the flavorings.\",\n \"For example, the time prior to mixing the flavoring with the brewing substance is nominal thereby virtually eliminating any degradation of the flavoring which might occur if the flavoring was exposed to air.\",\n \"A liquid flavor dispensing system 130 is illustrated diagrammatically in FIG.\",\n \"8.With this embodiment, the liquid flavor dispensing system dispenses a different liquid flavoring 132 in a controlled manner to several of the brewing assemblies of the beverage dispensing modules 32.The flavor can be dispensed to a water or liquid distribution system which controllably dispenses water to a beverage making substance, the beverage making substance or both.\",\n \"With reference to FIGS.\",\n \"8-10, the illustrated liquid flavor dispensing system 130 desirably includes a plurality of packages or containers 144a-144h, each of which retains a respective liquid flavor 132 therein.\",\n \"The containers 144 desirably are in controlled communication with respective beverage dispensing modules 32a-32d in any suitable manner.\",\n \"For example, a section of tubing or other passage 146 may connect each container 144 to a respective check valve 148.The check valve 148 is part of a respective solenoid or other form of pump device 150 coupled to the controller 76 via a respective line 152.A respective small diameter tube 154 extends from the solenoid pump 150 to a respective second check valve or flavor injection port 156 at a respective terminal end 158 of the tube 154.A respective sensor 160 can be coupled to the controller via a respective line 162 to monitor and detect a low level of the respective flavoring 132 in the respective container 144.As will be described in greater detail below with regard to sensing and monitoring, the status of the flavorings 132 can be reported by the controller 176.The check valves 148 and 156 at each end of each of the tubes 154 provide a sealed line to which the container 144 can be attached.\",\n \"Also, either alternatively or in addition to the use of the sensor 160, the flavor usage can be inferred based on information collected on a characteristic of the pump such as operation or usage for example, pump actuation.\",\n \"For example, a sensor may be associated with the pump and run time of the pump 150 could be monitored and a calculation made to infer the amount of flavorings remaining in a corresponding flavorings container 144.The system would provide a signal to the operator, such as “check-status” when the level of flavorings approaches, attains or drops below a predetermined level, for example 20% remaining.\",\n \"Such advance warning would give the operator the opportunity to change the flavorings container 144 before it runs out.\",\n \"The advance warning also helps assure that a reasonable margin of error in the inferred calculation is considered and accommodated for.\",\n \"In use, when a brewing cycle is initiated, the solenoid pump 150 can be activated simultaneous with the initiation of the brewing cycle of one of the beverage dispensing modules or shortly thereafter.\",\n \"A quantity, for example 9 ml for use in brewing ½ gallon of coffee, of liquid flavoring 132 is pumped from the container 144 through the tubing 154.The check valve or flavor injection port 156 is positioned in close proximity to a sprayhead 170 of the dispensing assembly 92.Dispensing of the flavor 132 into the water will allow the flavor to mix with the heated water for brewing substance during the saturation and extraction process.\",\n \"Generally, it is desirable to use the flavoring at the start of the cycle which will result in completely integrating the flavor during the brewing cycle.\",\n \"The process may be repeated with other beverage dispensing modules 32, 32a and, if desired, with other flavors.\",\n \"The flavor injection ports 156 may, for example, include a check valve of a type as produced by Smart Products.\",\n \"This is a positive pressure inverted check valve which requires positive pressure through the tube 154 to open the valve.\",\n \"Positioning of the injection port 156 in close proximity to the sprayhead 170 results in the benefit of clearing and rinsing the surface as a result of the water vapor and heat during a brewing cycle.\",\n \"Periodic manual cleaning of the sprayhead 170 and injection port 156 helps maintain a clean system.\",\n \"Desirably, because the injection port 156 is an inverted pressure check valve, physical contact with the tip of the injection port 156 through which the liquid 132 is dispensed does not result in dispensing of liquid 132.Rather, physical contact with the end of the injection port 156 causes the valve to close as opposed to open.\",\n \"The system also prevents the entry of atmosphere into the container 144 or line 154 thereby preventing contamination or degrading of the flavor or quality of the flavorings.\",\n \"This system provides a generally closed system which prevents the flavorings contact with the atmosphere and prevents contamination providing a generally aseptic environment.\",\n \"With reference to FIG.\",\n \"11, the liquid flavor dispensing system 130 as described with regard to FIGS.\",\n \"8-10 may be used with a soluble beverage system 176.In FIG.\",\n \"11, the liquid dispensing system dispenses liquid flavor into a hot water path 174 of a soluble or powdered beverage dispensing system 176.The devices used to dispense the liquid flavor are the same as those describe above with regard to FIG.\",\n \"8.As such, only the terminal ends 158a, 158b and 158c of multiple lines 154a, 154b, and 154c are shown.\",\n \"Similarly, the multiple injection ports 156a, 156b, and 156c are shown.\",\n \"The soluble beverage dispensing system 176 retains a quantity of powdered beverage mix (not shown) in a beverage dispensing container 178.The dispensing of soluble beverage material in powder form is generally known in the art.\",\n \"The container 178 includes a dispensing auger (not shown) which controllably dispenses powder by operation of a dispensing drive or motor 180 controlled by the controller 76a.\",\n \"Powdered beverage material is dispensed through the passage 182 into a mixing chamber 184.The hot water line 174 communicates with the mixing chamber 184 to combine with the powder.\",\n \"The combined powder and hot water flow into an agitating chamber 186 there below.\",\n \"A whipper motor 188 and agitating blade 190 further blend the powder and water combination for dispensing a beverage 192 through a nozzle 194.The flavor dispensing system 130 (see, FIG.\",\n \"8) controllably dispenses liquid flavor 132 from respective containers 144 using corresponding pumps via the lines 154a, 154b, and 154c.\",\n \"Either individual flavors or combinations of flavors may be dispensed into the hot water line.\",\n \"Such dispensing may be programmed in the controller 76a.\",\n \"Positioning of the injection ports 156a, 156b, 156c in the heated water path helps to assure that the liquid flavors are rinsed from the injection ports during and at the conclusion of the heated water dispensing cycle during a beverage dispensing cycle.\",\n \"It should be noted that generally these powdered beverage dispensing systems have a momentary hot water overrun at the end of the cycle during which powdered beverages dispense from the container 178.The slight overrun of heated water helps to assure that the mixing chamber 184, whipping chamber 186, and nozzle 194 are rinsed clean at the end of the cycle.\",\n \"The injection ports 156a, 156b and 156c have been positioned in the hot water line to provide the rinsing characteristics and benefits of the overrun of hot water.\",\n \"Furthermore, it is commonly known to have at least one daily rinse cycle in which heated water is dispensed through the assembly 176 to maintain cleanliness and sanitation of the system.\",\n \"During a brew cycle as shown in FIG.\",\n \"8, liquid flavor is generally preferably dispensed at the initiation of the brew cycle.\",\n \"As noted, this helps to ensure complete mixing and optimal extraction of the flavor from the beverage brewing substance 136.A predetermined volume of liquid flavor 132 can be dispensed into the brewing assembly 30 at the initiation of the brew cycle.\",\n \"In contrast, the soluble system 176 as shown in FIG.\",\n \"11 may have a variable quantity of beverage 192 dispensed therefrom.\",\n \"It is anticipated that multiple beverage sizes may be dispensed from the soluble system 176.As a result, the liquid flavoring 132 must be dispensed at a rate consistent with the concentration flavor profile or recipe, and powder also dispensed by the system.\",\n \"For example, if a consumer wishes to obtain a 12 ounce beverage the proper amount of liquid flavoring must be dispensed for a 12 ounce beverage.\",\n \"However, the consumer may be allowed to control whether they wish to have a full 12 ounces of beverage or if they decide to short the beverage resulting in, perhaps, only 10 ounces being dispensed.\",\n \"As such, the pump associated with the powdered beverage system 176 must be calibrated to dispense small quantities of liquid flavoring 132 on a per unit time basis to assure a desirable and consistent mixing of flavor with the powder.\",\n \"This will help assure that the desired flavor is achieved regardless of the quantity of beverage dispensed.\",\n \"The solenoid pump 150 used with the liquid flavor dispensing system 130 is a precision controlled metering pump.\",\n \"It is anticipated that other forms of the metering pump may be used with this system.\",\n \"However, in the present embodiment, the metering pump is controllably operated to dispense very small quantities of the liquid flavor 132.The frequency of the plunger rate, the stroke of the plunger as well as the duration of plunger operation the solenoid pump 150 can be controlled by the controller 76, 76a.\",\n \"As a result, very small quantities of highly concentrated liquid flavor can be dispensed into the beverages 140, 192 during the corresponding beverage preparation process.\",\n \"As such, precise quantities can be added to provide flexibility and tailoring of the resultant beverages.\",\n \"The system 130 can be calibrated using volumetric guidelines associated with various beverage recipes and flavor materials to match particular flavor preferences, for example for various demographics.\",\n \"As such, a franchise operation can custom blend and configure the flavor of beverages dispensed by the beverage dispensing system to meet and satisfy regional preferences.\",\n \"Additionally, if preferences change over time the system can be modified and calibrated to match those preferences.\",\n \"This adds further flexibility to the beverage system disclosed.\",\n \"With reference to FIGS.\",\n \"14-18, a powdered or soluble beverage dispensing system 199 is shown.\",\n \"While a soluable beverage system 199 has been shown and described as a powder dispensing system, it is envisioned that such a soluable system also includes a liquid concentrate system which dispenses a quality of liquid coffee concentrate for mixing with water or other dilution or reconsituting substance.\",\n \"Reference to a soluable beverage system is intended to be broadly interpreted.\",\n \"Such a beverage dispensing system has been generally and diagrammatically shown in FIG.\",\n \"11.With further reference to FIG.\",\n \"14, the container 178 is shown positioned inside a housing 200.The passage 182 coupled to the container 178 is shown positioned for dispensing into the mixing chamber 184 and communicating with the whipping chamber 186 and nozzle 194.A portion of the housing 200 in the form of a door 202 conceals the powdered beverage system 176 in the housing 200.Also, the liquid flavor dispensing system 130 as described hereinabove may be used with the powdered beverage or soluble beverage dispensing system 176 illustrated in FIGS.\",\n \"14-18.One of the potential difficulties involved with a soluble beverage dispensing system is that the container 178 must be removed periodically for refilling with powdered or soluble beverage substance as well as regular maintenance and cleaning.\",\n \"The container 178 may not be very heavy when it is empty because it is generally formed of a plastic material.\",\n \"However, once filled, it may contain 5 to 10 pounds of powder material.\",\n \"With the container 178 being positioned in an elevated location and containing a quantity of material, it may be difficult to lift the container into a desired position in the housing 200.The container 178 is positioned in an upper position in the housing 200 in order to benefit from gravity assistance when dispensing powdered beverage therefrom to the mixing chamber 84.Additionally, it is important to align the dispensing passage 182 with the housing 184 to help assure that when powdered beverage material is dispensed from the container 178 it flows into the proper path for mixing with water.\",\n \"As such, it would be desirable to provide an apparatus and system for facilitating improved ease of removal and replacement of the powdered beverage container 178 relative to the housing 200.As shown in FIGS.\",\n \"14-18, a container positioning assembly or container positioner 220 in the form of a slide-out support or shelf is provided.\",\n \"The container positioning assembly 220 includes a pair of handles 222 which can be grasped (see FIG.\",\n \"15) by a user to extract the container 178 attached to the assembly 220 for removing it from the housing 200.As shown in FIG.\",\n \"15, the assembly 220 allows the container 178 to be pulled straight out of the housing 200 thereby allowing disengagement of the passage 182 relative to the mixing chamber 184.Movement of the assembly 220 also replaces the container 178 in a desired position to align the dispensing passage 182 with the mixing chamber 184.As can be seen in FIGS.\",\n \"15-17, the container 178 has been removed from the housing 200 in order to refill the container 178 but does not have to be removed from the shelf 250 of the assembly 220.Support rails 226 are positioned on each side of container 178 and are engaged with side walls 228 of the housing 200 (FIG.\",\n \"15).\",\n \"Each of the guide rails 226 includes a pair of tracking slots 230, 232 which are engaged with corresponding upper 234 and lower 236 guide rollers.\",\n \"A support roller 240 is positioned along an exterior edge 242 of the guide 226 to further help facilitate smooth and controlled movement of the assembly 220.Support rollers 240 and guide rollers 234, 236 are mounted on the sidewalls of the housing 200.The guides 226 are connected by a cross extending shelf 250 on which the container 178 is positioned.\",\n \"The combination of the guide rails 226 and rollers help the assembly 220 slide outwardly and inwardly relative to the housing 200.FIG.\",\n \"16 shows only the portion of the guide rails 226 extending past the edge of container 178.The guide rollers 240, 234, 236 associated with the guides 226 prevent tipping of the shelf 250 and associated container 178.When the assembly 220 is stowed in the housing 200 the shelf 250 rests on a corresponding surface 252 of the housing 200.As can be seen in FIGS.\",\n \"15-17, the guides 226 include a generally horizontal portion 254 and a vertically oriented portion 256.The shelf 250 and container 178 are associated with the generally horizontal portion 254.The generally vertical portion 256 helps to allow forward movement as well as downward movement of the assembly 220 relative to the housing 200.The guide including the portion 254, 256 help facilitate movement of the container into and out of the housing 200 and forward and downward movement.\",\n \"In the embodiment as shown, the assembly facilitates positioning of the container 178 approximately 6-8 inches downwardly from its stored position in the housing 200.This lowering of the container 178 is beneficial in that it allows for easy filling of the container.\",\n \"Also, by fixing the container to the shelf 250 and the assembly 200 it prevents spilling or otherwise tipping of the container 178 during filling or other processes.\",\n \"This can result in savings in terms of lost beverage material as well as cleanup costs and associated complications.\",\n \"In the stored position, detents 260, 262 are provided on the guides 230, 232, respectively.\",\n \"The detents 260, 262 allow the rollers 234, 236 respectively, to be engaged therewith to prevent casual disengagement of the assembly 220 from the housing 200.This helps to lock or retain the assembly 220 in a desired stored or stowed position in the housing 200.As the assembly 220 is moved outwardly from the housing 200, the track guides 234, 236 track along the corresponding channels 230, 232 generally horizontally along the horizontal portion 254.As such, the container 278 attached to the shelf 250 sides forwardly and outwardly from the housing.\",\n \"At a position where the generally horizontal portion 254 transitions into the partially vertical portion 256 (elbow 270) the guides allow the assembly to slide downwardly and forwardly.\",\n \"In the extended most position the guides 234, 236 engage corresponding extended detents 272, 274, respectively.\",\n \"These detent positions 272, 274 help retain the assembly 220 in the outward or extended most position.\",\n \"The assembly 220 also includes an assist system 280 which includes cables 282 and biasing members to springs 284 forming a cable-biasing assembly help facilitate removal of and replacement of the container 178 relative to the housing 200.With reference to FIG.\",\n \"18, the assist system 280 includes a spring 284 attached at one 286 to a back portion of the housing and at a second end to a first pulley 288.The cable 282 is attached at a first end 290 to a corresponding portion of the housing and extends through the first pulley 288 and up and around a second pulley 292.The second end 294 of the cable 282 attaches to a corresponding portion of the guides 226 at attachment point 296.Attachment of the cable 282 to the guide 226 is shown in FIG.\",\n \"16.The assist assembly 288 helps further facilitate ease of removal and replacement of the container 178 relative to the housing 200.For example, when the container 178 is removed from the housing on the assembly 220 the spring 284 stores energy and is stretched as the cables 282 are extended.\",\n \"The cables 282 are extended as a result of the forward and downward movement of the guides 226.Extension of the springs 284 expends the stored energy and imparts a return force in the springs which helps reduce the force required to return the container 178 to the housing 200.This is particularly useful when an empty container is removed from the housing, filled with powdered material and then returned.\",\n \"Returning the filled container 178 normally would require additional effort on the part of the operator and as such the return forces in the assist system 280 help make the return easier.\",\n \"Turning now to FIGS.\",\n \"19 and 20, the beverage system also includes a communications system or means 300 for communicating 302 between a corresponding beverage brewing, beverage making apparatus or beverage dispensing modules 32, 32a and a receiver or base station 304.Each of the modules 32, 32a includes a controller 76, 76a which is centralized for use with multiple modules or dedicated to an individual module.\",\n \"The controller collects information about the module or modules.\",\n \"For purposes of this present discussion, we will refer to the controller as being dedicated to multiple modules and receiving information from each individual module associated therewith.\",\n \"The many modules associated with a single control also provides the added benefit of the modular assembly as described hereinabove.\",\n \"It should also be noted that the details of the circuitry used to achieve the communication system are illustrated in the detailed schematics provided in FIGS.\",\n \"21-28 and all corresponding sub-portions thereof.\",\n \"For example, FIG.\",\n \"21 is a general schematic of the overall system.\",\n \"FIGS.\",\n \"22-28 provide details about the general system shown in FIG.\",\n \"21.FIG.\",\n \"21 has been noted with cross reference to the other figures which provide details about the portions of the circuitry shown generally in FIG.\",\n \"21.The controllers 76, 76a include the link 302 which connects the controller to the corresponding receiver or base station 304.In the present embodiment as diagrammatically illustrated in FIGS.\",\n \"19 and 20, the controllers 76, 76a communicate with the receiver 304 by way of an RF link 302.Generally, the controllers are remote from the receiver and use the Link 302 and the associated means for linking to communicate the collected information about the beverage making apparatus.\",\n \"Antennae 306, 308 are provided on the controllers 76, 76a and the corresponding receivers 304 respectively.\",\n \"It should be noted that the communication link 302 may be in any one of a variety of forms such as hardwired physical point-to-point link, optical lines, light wave, ultrasonic, infrared or any other form of communication link between one or more devices.\",\n \"The receiver 304 receives information from the controller or various components of the modules 32, 32a to identify information relating to the modules and operation of the modules.\",\n \"The receiver 304 may be in the form of a monitoring unit positioned in an appropriate location relative to an operator or attendant who has responsibility for or otherwise maintains the modules 32, 32a.\",\n \"The receiver 304 includes a means for displaying display 305 and can provide visual auditor or other information about numerous conditions.\",\n \"The display 304 allows a form of the collected information about the beverage making apparatus to be displayed at the receiver.\",\n \"The visual display may include, but is not limited to lights, text, symbolic images (i.e., dispenser “full” showing a colored or shaded dispenser and a dispenser “empty” showing an unshaded dispenser), and mechanical devices that tare operated or other visual displays.\",\n \"An auditory display may include, but is not limited to speech information, alarms, tones or other signals that van be heard.\",\n \"Tactile displays are also contemplated in the form of a vibrating surface, vibrating device worn by the operator or other means for letting the operator or other means for letting the operator know about he collected information.\",\n \"For example, the following is a list of the messages or conditions which can be displayed: Receiver fault; Please wait attempting communication; Communication failure, coffee module #; Communication fault, soluble module #; Flavor [type] low, coffee module #; Flavor [type] low, soluble module #; Hopper low, soluble module #; Hopper empty, soluble module #; Coffee empty, coffee module #; Coffee low, coffee module #; Freshness expired, coffee module #; Dumping in minutes, coffee unit #; Server removed, coffee module #; Refill too long; Temperature probe open; Temperature probe short; Heating too long; Over flow safety; Coffee stations all OK; and Soluble stations all OK. For example, in an convenience store setting, multiple modules 32, 32a may be generally remotely positioned in one portion of the store, for example, a rearward location of the store, if desired.\",\n \"The operator of the beverage system may also be the same person responsible for stocking of the equipment, and register activities.\",\n \"As such, one person is required to operate and maintain many components of the convenience store facility.\",\n \"The receiver 304 may be positioned at the cash register so that while the attendant is operating the cash register he may also be notified of matters that require his attention at the modules 32, 32a.\",\n \"For example, if a module 32a indicates that a container 68 is empty the controller 76 will communicate with the receiver 304 to indicate this condition.\",\n \"The receiver may also be configured with audible alarms and visual displays to provide additional information.\",\n \"For example, a display may identify which module and the specific condition associated with the module which has provided a signal to the receiver 304.The operator can then make a decision how to use that information relative to his responsibilities at the cash register.\",\n \"If an emergency requires immediate attention he can secure the cash register and address the emergency while attending to the module 32.Alternatively, if it is a slow period in the store and a module is indicating that a container 68 is empty he can put off attending to this condition until he completes one or more transactions in queue.\",\n \"The signal communication between the controller 76 and the receiver 304 may be a single path signal or in the form of a multiple path verification signal.\",\n \"For example, as shown in FIGS.\",\n \"19 and 20 a first signal 310 is transmitted from the controller 76 to the receiver 304.This initial communication may report the status of a condition at the module 32.The receiver 304 can then return a copy of the information 312 to the controller 76.If the controller confirms the information received (312) it can then send a return confirmation 314 that the information is correct.\",\n \"This is a useful communication link in the present situation due to the presence of electrical noise and other interference within other beverage system applications.\",\n \"For example, each of the components associated with the modules 32, 32a may provide some degree of electrical or other noise.\",\n \"Additionally, other systems within the beverage system setting may be producing RF or other signals which could create interference.\",\n \"As such, the present embodiment providing the verification communication links 310-314 helps facilitate and assure accurate and timely communication.\",\n \"In the event of excessive interference such as from patrons of the store using telephone or RF communication devices, the controller 76 can continue to periodically send a message 310 to the receiver 304 until the verification signal 312 is received.\",\n \"While preferred embodiments are disclosed, illustrated and described, it is envisioned that those skilled in the art may devise various modifications and equivalents without departing from the spirit and scope of the disclosure as recited in the following claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476928"}}},{"rowIdx":984,"cells":{"text":{"kind":"list like","value":[["Windscreen wiper device","A windscreen wiper device comprising an elastic, elongated carrier element, as well as an elongated wiper blade of a flexible material, which can be placed in abutment with a windscreen to be wiped, which wiper blade includes opposing longitudinal grooves on its longitudinal sides, in which grooves space-apart longitudinal strips of the carrier element are disposed, wherein neighboring ends of said longitudinal strips are interconnected by a respective connecting piece, which windscreen wiper device comprises a connecting device for an oscillating wiper arm, as well as a spoiler, a special feature of which is that the spoiler as a separate constructional element is detachably connected to the wiper blade."],["1-12.","(canceled) 13.A windscreen wiper device comprising: an elastic, elongated carrier element; an elongated wiper blade of a flexible material, which can be placed in abutment with a windscreen to be wiped, said wiper blade extending between longitudinally opposite ends and including opposing longitudinal grooves formed on longitudinal sides thereof, said carrier element including spaced-apart longitudinal strips disposed in said grooves and interconnected by a respective connecting piece at opposite neighboring end of said strips; a connecting device for an oscillating wiper arm; and as a separate element from that of said wiper blade and being detachable connected to said wiper blade and extending from said connecting device substantially to said ends of said wiper blade.","14.A windscreen wiper device according to claim 13, wherein said spoiler comprises separate subspoiler pieces disposed on opposite sides of said connecting piece.","15.A windscreen wiper device according to claim 13, wherein said spoiler comprises clamping features which detachably engage longitudinal sides of said longitudinal strips that face away from each other.","16.A windscreen wiper device according to claim 13, wherein said spoiler comprises clamping features which detachably engage longitudinal edges of said wiper blade that face away from each other.","17.A windscreen wiper device according to claim 16, wherein said spoiler and said clamping features are integral.","18.A windscreen wiper device according to claim 13, wherein said connecting pieces are clamping members, which form separate constructional elements.","19.A windscreen wiper device according to claim 18, wherein said connecting pieces are form-locked or force-locked to the adjacent ends of said longitudinal strips.","20.A windscreen wiper device according to claim 13, wherein said connecting pieces are in one piece with said longitudinal strips.","21.A windscreen wiper device according to claim 13, wherein said longitudinal strips are made of spring band material.","22.A windscreen wiper device according to claim 13, wherein said connecting device comprises clamping members, which engage longitudinal sides of said longitudinal strips that face away from each other.","23.A windscreen wiper device according to claim 22, wherein said connecting device and said clamping members are made in one piece.","24.A method for manufacturing a windscreen wiper device comprising preparing a wiper blade of flexible material which can be placed into abutment with a windscreen to be cleaned having opposing longitudinal grooves formed in longitudinal sides of the wiper blade, positioning longitudinal strips of a carrier element in the grooves of the wiper blade in spaced-apart relationship, and connecting neighboring ends of, the strips, with respective connecting pieces; and fabricating a spoiler as a separate constructional element from that of the wiper blade, and detachably mounting the spoiler on the wiper blade."],["The invention relates to a windscreen wiper device comprising an elastic, elongated carrier element, as well as an elongated wiper blade of a flexible material, which can be placed in abutment with a windscreen to be wiped, which wiper blade includes opposing longitudinal grooves on its longitudinal sides, in which grooves spaced-apart longitudinal strips of the carrier element are disposed, wherein neighboring ends of said longitudinal strips are interconnected by a respective connecting piece, which windscreen wiper device comprises a connecting device for an oscillating wiper arm, as well as a spoiler.","Such a spoiler is also called an “air deflector”.","Such a windscreen wiper device is generally known.","The prior art windscreen wiper device is in particular designed as a “yokeless” wiper device, wherein use is no longer made of several yokes pivotally connected to each other, but wherein the wiper blade is biased by the carrier element, as a result of which it exhibits a specific curvature.","The spoiler and the wiper blade of the known windscreen wiper device are made in one piece.","Windscreen wiper devices actually used on cars fitted with windscreens exhibiting some degree of curvature are sometimes subjected to a lifting effect at high speeds, which prevents the windscreen from being wiped correctly.","This lifting effect is due to the following aerodynamic phenomenon.","Streams of air along the car tend to escape vertically upwards along the axis of the car.","Near the sides of the curved windscreen, however, the streams of air tend to pass horizontally along the sides.","These streams of air create, mainly in the zones in which the windscreen wiper device meets them perpendicularly, a reduced pressure above the windscreen wiper device and an increased pressure between the windscreen and the windscreen wiper device.","The resulting pressure, in spite of the mechanical pressure applied by the arm of the windscreen wiper device, causes the latter to lift away towards the front of the windscreen, thereby deteriorating the wiping pattern at higher driving speeds.","A disadvantage of the known windscreen wiper device is that the spoiler and the wiper blade are made in one piece: every car has its own specific aerodynamic characteristics, so that a specific windshield wiper with an integrated spoiler has to be designed and manufactured for each type of car.","Obviously this requires the use of complex machinery, tools, with all the expenses involved.","A further drawback is that replacement (because of wear) of the wiper blade automatically implies the replacement of the spoiler.","It is an object of the invention to obviate these disadvantages, in the sense that at minimum cost a windshield wiper device comprising a spoiler is proposed which can be mounted on all windshield wipers of different cars, without the use of complex machinery and additional tools being required.","It is noted that the present invention is not restricted to windscreen wiper devices for cars, but that it also relates to windscreen wiper devices for rail coaches and all other (fast) vehicles.","In order to accomplish that objective, a windscreen wiper device of the kind referred to in the introduction is characterized in that the spoiler as a separate constructional element is detachably connected to the wiper blade.","In particular said spoiler comprises on opposite sides of the connecting device a subspoiler being detachably connected to the wiper blade.","The use of two subspoilers ensures a great rigidity of the spoiler.","In one preferred embodiment of a windscreen wiper device according to the invention, said spoiler comprises clamping means which engage round longitudinal sides of said longitudinal strips that face away from each other.","In another preferred embodiment said spoiler comprises clamping means which engage round longitudinal edges of said wiper blade that face away from each other.","Preferably said spoiler and said clamping means are made in one piece.","In another preferred embodiment of a windscreen wiper device according to the invention, said connecting pieces are clamping members, which form separate constructional elements.","In particular, said connecting pieces are form-locked or force-locked to the adjacent ends of the longitudinal strips.","In another preferred embodiment of a windscreen wiper device according to the invention, said connecting pieces are in one piece with said longitudinal strips.","In another preferred embodiment of a windscreen wiper device according to the invention, at least said longitudinal strips are made of spring band material, preferably steel.","In another preferred embodiment of a windscreen wiper device according to the invention, said connecting device comprises clamping members, which engage round longitudinal sides of said longitudinal strips that face away from each other.","In particular, said connecting device and said clamping members are made in one piece.","The invention furthermore relates to a method for manufacturing a windscreen wiper device according to the invention, wherein opposing longitudinal grooves are formed in the longitudinal sides of an elongated wiper blade of a flexible material, which can be placed in abutment with a windscreen to be wiped, in which grooves longitudinal strips of a carrier element are subsequently fitted in spaced-apart relationship, wherein neighboring ends of said longitudinal strips are interconnected by a respective connecting piece, characterized in that a spoiler as a separate constructional element is detachably fitted onto the wiper blade.","The invention will now be explained in more detail with reference to figures illustrated in a drawing, wherein: FIG.","1 shows diagrammatically the flow of streams of air along the windscreen of a car; FIG.","2 is a diagram of a standard windscreen wiper device in a position parallel to the streams of air; FIG.","3 is a cross-section of the same windscreen wiper device, but in a position at right angles to the streams of air; FIG.","4 illustrates the forces due to the tangential streams of air acting on the standard windscreen wiper blade; FIGS.","5 and 6 show a schematical and a perspective view, respectively, of a windscreen wiper device according to two preferred embodiments of the invention; and FIG.","7 is a total perspective, schematic view of the windscreen wiper device of FIGS.","5 and 6.FIG.","1 shows a perspective view of the front portion of a car provided with a curved windscreen A, together with the direction of flow of the streams of air, the latter passing the vehicle upwards and along the sides.","In a zone in which a windscreen wiper device of the usual standard form is positioned parallel to the streams of air, as shown in FIG.","2, nothing unusual occurs; on the other hand, in a zone as shown in FIG.","3, in which the streams of air flow practically at right angles to the direction of the windscreen wiper device, this air cannot flow along the glass because of the wiper blade, and thus creates an increase in pressure (indicated by the + sign ).","Such an increased pressure also exists in the region of the hollow profile comprised between the flanges of the frame C directed towards the windscreen.","These flanges create further obstacles to the flow of air and assist in maintaining the increased pressure.","In addition, the streams of air which pass round the windscreen wiper device towards the upstream side tend to move away from the surface and create a reduced pressure zone (indicated by the − sign), as is well known.","These two forces Fx and Fz (which can be compared with the drag and the lift acting on a wing section) have a resultant R which tends to lift the windscreen wiper device against the mechanical pressure applied by the arm (see FIG.","4).","FIGS.","5 and 6 show a preferred variant of a windscreen wiper device 1 according to the invention.","Said windscreen wiper device is built up of an elastomeric wiper blade 2, in the longitudinal sides of which opposing longitudinal grooves 3 are formed, and of longitudinal strips 4 made of spring band steel, which are fitted in said longitudinal grooves 3.Said strips 4 form a flexible carrier element for the rubber wiper blade 2, as it were, which is thus biassed in a curved position (the curvature in operative position being that of a windscreen to be wiped).","Neighbouring ends 5 of strips 4 are interconnected on either side of the windscreen wiper device 1 by means of connecting pieces 6 functioning as clamping members.","In this embodiment, the connecting pieces 6 are separate constructional elements, which may be form-locked as well as force-locked to the ends 5 of strips 4.In another preferred variant, said connecting pieces 6 are in one piece with the strips 4 made of spring band steel.","In the latter case said connecting pieces form transverse bridges for the strips 4, as it were.","The windscreen wiper device 1 is furthermore built up of a connecting device 7 of plastic material for an oscillating wiper arm 8.Connecting device 7 comprises clamping members 9 that are integral therewith, which engage round longitudinal sides 10 of the strips 4 that face away from each other, as a result of which the connecting device 7 is firmly attached to the unit consisting of wiper blade 2 and strips 4.The oscillating wiper arm 8 is pivotally connected to the connecting device 7 about a pivot axis near one end.","Both preferred embodiments of FIGS.","5 and 6 according to the invention comprise a spoiler 11 including two separate subspoilers 12 being detachably connected to the wiper blade 2 on opposite sides of the connecting device 7.The subspoilers 12 are made of an elastomeric material, rubber for example, and they are provided with clamping means 13 made in one piece therewith.","In the preferred embodiment of FIG.","5 the clamping means 13 engage round longitudinal sides 10 of the longitudinal strips 4 that face away from each other, whereas in the embodiment of FIG.","6 the clamping means 13 engage around longitudinal laterally extending edges 14 of the wiper blade 2 that face away from each other.","Of course, it is also possible for the clamping means to engage round other longitudinal laterally extending edges 15 of the wiper blade 2 that face away from each other.","FIG.","7 shows a total perspective, schematic view of the windscreen wiper device of FIGS.","5 and 6, wherein corresponding parts have been designated with the same reference numerals.","The invention is not restricted to the variants as shown in the drawing, but it also extends to other embodiments that fall within the scope of the appended claims."]],"string":"[\n [\n \"Windscreen wiper device\",\n \"A windscreen wiper device comprising an elastic, elongated carrier element, as well as an elongated wiper blade of a flexible material, which can be placed in abutment with a windscreen to be wiped, which wiper blade includes opposing longitudinal grooves on its longitudinal sides, in which grooves space-apart longitudinal strips of the carrier element are disposed, wherein neighboring ends of said longitudinal strips are interconnected by a respective connecting piece, which windscreen wiper device comprises a connecting device for an oscillating wiper arm, as well as a spoiler, a special feature of which is that the spoiler as a separate constructional element is detachably connected to the wiper blade.\"\n ],\n [\n \"1-12.\",\n \"(canceled) 13.A windscreen wiper device comprising: an elastic, elongated carrier element; an elongated wiper blade of a flexible material, which can be placed in abutment with a windscreen to be wiped, said wiper blade extending between longitudinally opposite ends and including opposing longitudinal grooves formed on longitudinal sides thereof, said carrier element including spaced-apart longitudinal strips disposed in said grooves and interconnected by a respective connecting piece at opposite neighboring end of said strips; a connecting device for an oscillating wiper arm; and as a separate element from that of said wiper blade and being detachable connected to said wiper blade and extending from said connecting device substantially to said ends of said wiper blade.\",\n \"14.A windscreen wiper device according to claim 13, wherein said spoiler comprises separate subspoiler pieces disposed on opposite sides of said connecting piece.\",\n \"15.A windscreen wiper device according to claim 13, wherein said spoiler comprises clamping features which detachably engage longitudinal sides of said longitudinal strips that face away from each other.\",\n \"16.A windscreen wiper device according to claim 13, wherein said spoiler comprises clamping features which detachably engage longitudinal edges of said wiper blade that face away from each other.\",\n \"17.A windscreen wiper device according to claim 16, wherein said spoiler and said clamping features are integral.\",\n \"18.A windscreen wiper device according to claim 13, wherein said connecting pieces are clamping members, which form separate constructional elements.\",\n \"19.A windscreen wiper device according to claim 18, wherein said connecting pieces are form-locked or force-locked to the adjacent ends of said longitudinal strips.\",\n \"20.A windscreen wiper device according to claim 13, wherein said connecting pieces are in one piece with said longitudinal strips.\",\n \"21.A windscreen wiper device according to claim 13, wherein said longitudinal strips are made of spring band material.\",\n \"22.A windscreen wiper device according to claim 13, wherein said connecting device comprises clamping members, which engage longitudinal sides of said longitudinal strips that face away from each other.\",\n \"23.A windscreen wiper device according to claim 22, wherein said connecting device and said clamping members are made in one piece.\",\n \"24.A method for manufacturing a windscreen wiper device comprising preparing a wiper blade of flexible material which can be placed into abutment with a windscreen to be cleaned having opposing longitudinal grooves formed in longitudinal sides of the wiper blade, positioning longitudinal strips of a carrier element in the grooves of the wiper blade in spaced-apart relationship, and connecting neighboring ends of, the strips, with respective connecting pieces; and fabricating a spoiler as a separate constructional element from that of the wiper blade, and detachably mounting the spoiler on the wiper blade.\"\n ],\n [\n \"The invention relates to a windscreen wiper device comprising an elastic, elongated carrier element, as well as an elongated wiper blade of a flexible material, which can be placed in abutment with a windscreen to be wiped, which wiper blade includes opposing longitudinal grooves on its longitudinal sides, in which grooves spaced-apart longitudinal strips of the carrier element are disposed, wherein neighboring ends of said longitudinal strips are interconnected by a respective connecting piece, which windscreen wiper device comprises a connecting device for an oscillating wiper arm, as well as a spoiler.\",\n \"Such a spoiler is also called an “air deflector”.\",\n \"Such a windscreen wiper device is generally known.\",\n \"The prior art windscreen wiper device is in particular designed as a “yokeless” wiper device, wherein use is no longer made of several yokes pivotally connected to each other, but wherein the wiper blade is biased by the carrier element, as a result of which it exhibits a specific curvature.\",\n \"The spoiler and the wiper blade of the known windscreen wiper device are made in one piece.\",\n \"Windscreen wiper devices actually used on cars fitted with windscreens exhibiting some degree of curvature are sometimes subjected to a lifting effect at high speeds, which prevents the windscreen from being wiped correctly.\",\n \"This lifting effect is due to the following aerodynamic phenomenon.\",\n \"Streams of air along the car tend to escape vertically upwards along the axis of the car.\",\n \"Near the sides of the curved windscreen, however, the streams of air tend to pass horizontally along the sides.\",\n \"These streams of air create, mainly in the zones in which the windscreen wiper device meets them perpendicularly, a reduced pressure above the windscreen wiper device and an increased pressure between the windscreen and the windscreen wiper device.\",\n \"The resulting pressure, in spite of the mechanical pressure applied by the arm of the windscreen wiper device, causes the latter to lift away towards the front of the windscreen, thereby deteriorating the wiping pattern at higher driving speeds.\",\n \"A disadvantage of the known windscreen wiper device is that the spoiler and the wiper blade are made in one piece: every car has its own specific aerodynamic characteristics, so that a specific windshield wiper with an integrated spoiler has to be designed and manufactured for each type of car.\",\n \"Obviously this requires the use of complex machinery, tools, with all the expenses involved.\",\n \"A further drawback is that replacement (because of wear) of the wiper blade automatically implies the replacement of the spoiler.\",\n \"It is an object of the invention to obviate these disadvantages, in the sense that at minimum cost a windshield wiper device comprising a spoiler is proposed which can be mounted on all windshield wipers of different cars, without the use of complex machinery and additional tools being required.\",\n \"It is noted that the present invention is not restricted to windscreen wiper devices for cars, but that it also relates to windscreen wiper devices for rail coaches and all other (fast) vehicles.\",\n \"In order to accomplish that objective, a windscreen wiper device of the kind referred to in the introduction is characterized in that the spoiler as a separate constructional element is detachably connected to the wiper blade.\",\n \"In particular said spoiler comprises on opposite sides of the connecting device a subspoiler being detachably connected to the wiper blade.\",\n \"The use of two subspoilers ensures a great rigidity of the spoiler.\",\n \"In one preferred embodiment of a windscreen wiper device according to the invention, said spoiler comprises clamping means which engage round longitudinal sides of said longitudinal strips that face away from each other.\",\n \"In another preferred embodiment said spoiler comprises clamping means which engage round longitudinal edges of said wiper blade that face away from each other.\",\n \"Preferably said spoiler and said clamping means are made in one piece.\",\n \"In another preferred embodiment of a windscreen wiper device according to the invention, said connecting pieces are clamping members, which form separate constructional elements.\",\n \"In particular, said connecting pieces are form-locked or force-locked to the adjacent ends of the longitudinal strips.\",\n \"In another preferred embodiment of a windscreen wiper device according to the invention, said connecting pieces are in one piece with said longitudinal strips.\",\n \"In another preferred embodiment of a windscreen wiper device according to the invention, at least said longitudinal strips are made of spring band material, preferably steel.\",\n \"In another preferred embodiment of a windscreen wiper device according to the invention, said connecting device comprises clamping members, which engage round longitudinal sides of said longitudinal strips that face away from each other.\",\n \"In particular, said connecting device and said clamping members are made in one piece.\",\n \"The invention furthermore relates to a method for manufacturing a windscreen wiper device according to the invention, wherein opposing longitudinal grooves are formed in the longitudinal sides of an elongated wiper blade of a flexible material, which can be placed in abutment with a windscreen to be wiped, in which grooves longitudinal strips of a carrier element are subsequently fitted in spaced-apart relationship, wherein neighboring ends of said longitudinal strips are interconnected by a respective connecting piece, characterized in that a spoiler as a separate constructional element is detachably fitted onto the wiper blade.\",\n \"The invention will now be explained in more detail with reference to figures illustrated in a drawing, wherein: FIG.\",\n \"1 shows diagrammatically the flow of streams of air along the windscreen of a car; FIG.\",\n \"2 is a diagram of a standard windscreen wiper device in a position parallel to the streams of air; FIG.\",\n \"3 is a cross-section of the same windscreen wiper device, but in a position at right angles to the streams of air; FIG.\",\n \"4 illustrates the forces due to the tangential streams of air acting on the standard windscreen wiper blade; FIGS.\",\n \"5 and 6 show a schematical and a perspective view, respectively, of a windscreen wiper device according to two preferred embodiments of the invention; and FIG.\",\n \"7 is a total perspective, schematic view of the windscreen wiper device of FIGS.\",\n \"5 and 6.FIG.\",\n \"1 shows a perspective view of the front portion of a car provided with a curved windscreen A, together with the direction of flow of the streams of air, the latter passing the vehicle upwards and along the sides.\",\n \"In a zone in which a windscreen wiper device of the usual standard form is positioned parallel to the streams of air, as shown in FIG.\",\n \"2, nothing unusual occurs; on the other hand, in a zone as shown in FIG.\",\n \"3, in which the streams of air flow practically at right angles to the direction of the windscreen wiper device, this air cannot flow along the glass because of the wiper blade, and thus creates an increase in pressure (indicated by the + sign ).\",\n \"Such an increased pressure also exists in the region of the hollow profile comprised between the flanges of the frame C directed towards the windscreen.\",\n \"These flanges create further obstacles to the flow of air and assist in maintaining the increased pressure.\",\n \"In addition, the streams of air which pass round the windscreen wiper device towards the upstream side tend to move away from the surface and create a reduced pressure zone (indicated by the − sign), as is well known.\",\n \"These two forces Fx and Fz (which can be compared with the drag and the lift acting on a wing section) have a resultant R which tends to lift the windscreen wiper device against the mechanical pressure applied by the arm (see FIG.\",\n \"4).\",\n \"FIGS.\",\n \"5 and 6 show a preferred variant of a windscreen wiper device 1 according to the invention.\",\n \"Said windscreen wiper device is built up of an elastomeric wiper blade 2, in the longitudinal sides of which opposing longitudinal grooves 3 are formed, and of longitudinal strips 4 made of spring band steel, which are fitted in said longitudinal grooves 3.Said strips 4 form a flexible carrier element for the rubber wiper blade 2, as it were, which is thus biassed in a curved position (the curvature in operative position being that of a windscreen to be wiped).\",\n \"Neighbouring ends 5 of strips 4 are interconnected on either side of the windscreen wiper device 1 by means of connecting pieces 6 functioning as clamping members.\",\n \"In this embodiment, the connecting pieces 6 are separate constructional elements, which may be form-locked as well as force-locked to the ends 5 of strips 4.In another preferred variant, said connecting pieces 6 are in one piece with the strips 4 made of spring band steel.\",\n \"In the latter case said connecting pieces form transverse bridges for the strips 4, as it were.\",\n \"The windscreen wiper device 1 is furthermore built up of a connecting device 7 of plastic material for an oscillating wiper arm 8.Connecting device 7 comprises clamping members 9 that are integral therewith, which engage round longitudinal sides 10 of the strips 4 that face away from each other, as a result of which the connecting device 7 is firmly attached to the unit consisting of wiper blade 2 and strips 4.The oscillating wiper arm 8 is pivotally connected to the connecting device 7 about a pivot axis near one end.\",\n \"Both preferred embodiments of FIGS.\",\n \"5 and 6 according to the invention comprise a spoiler 11 including two separate subspoilers 12 being detachably connected to the wiper blade 2 on opposite sides of the connecting device 7.The subspoilers 12 are made of an elastomeric material, rubber for example, and they are provided with clamping means 13 made in one piece therewith.\",\n \"In the preferred embodiment of FIG.\",\n \"5 the clamping means 13 engage round longitudinal sides 10 of the longitudinal strips 4 that face away from each other, whereas in the embodiment of FIG.\",\n \"6 the clamping means 13 engage around longitudinal laterally extending edges 14 of the wiper blade 2 that face away from each other.\",\n \"Of course, it is also possible for the clamping means to engage round other longitudinal laterally extending edges 15 of the wiper blade 2 that face away from each other.\",\n \"FIG.\",\n \"7 shows a total perspective, schematic view of the windscreen wiper device of FIGS.\",\n \"5 and 6, wherein corresponding parts have been designated with the same reference numerals.\",\n \"The invention is not restricted to the variants as shown in the drawing, but it also extends to other embodiments that fall within the scope of the appended claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476929"}}},{"rowIdx":985,"cells":{"text":{"kind":"list like","value":[["Tunnel first safety device","In order to increase the safety of tunnels when a fire occurs, the tunnel is divided into sections (A) which are then separated from each other by curtains (1).","Each section (A) is provided with at least one fire sensor (2) which closes said curtains (1) in said section (A) if a fire breaks out."],["1.A safety device for fires in tunnels, characterized in that the tunnel has portions (A) that are separable from each other by curtains (1), each portion (A) being provided with at least one fire sensor (2) that causes the curtains (1) of the portion (A) to be closed in the event of a fire.","2.A safety device according to claim 1, characterized in that the curtains (1) are closed by a motor that is driven by the fire sensor.","(2) 3.A safety device according to claim 1, characterized in that the curtains (1) are drawn across the roadway (9) for closing.","4.A safety device according to claim 3, characterized in that the drawn up curtains (1) are disposed on the tunnel wall.","5.A safety device according to claim 4, characterized in that the drawn up curtains (1) are disposed in a recess (5) in the tunnel wall.","6.A safety device according to claim 1, characterized in that the drawn up curtains (1) are disposed in spaces closable by curtain flaps (6), the curtain flaps (6) being opened in the event of a fire.","7.A safety device according to claim 1, characterized in that the curtains (1) consist of strips (11) extending from above to below.","8.A safety device according to claim 7, characterized in that the strips (11) overlap in the extended state of the curtains (1).","9.A safety device according to claim 1, characterized in that the curtains are formed to be fireproof.","10.A safety device according to claim 1, characterized in that each curtain (1) consists of at least two curtain parts (3, 4) on each tunnel side that are drawn in the direction of the other tunnel side for closing.","11.A safety device according to claim 1, characterized in that each curtain consists in the closed state of two portions (3, 4) spaced one behind the other.","12.A safety device according to claim 1, characterized in that the curtains are guided on slide rails (7) provided in the area of the tunnel ceiling (8).","13.A safety device according to claim 1, characterized in that the curtains (1) are formed in such a way as to close off the upper tunnel area more tightly than the lower tunnel area.","14.A safety device according to claim 1, characterized in that each tunnel portion (A) is provided with at least one device for vitiated air suction.","15.A safety device according to claim 14, characterized in that the vitiated air suction device has a connection to the vitiated air duct (12) of the tunnel, said connection being closable with a shut-off device (14).","16.A safety device according to claim 15, characterized in that in the event of a fire the shut-off device (14) of the vitiated air device in the portion (A) with the fire is opened by the fire sensor (2) and the shut-off devices of the vitiated air suction devices of the other portions are at least partly closed.","17.A safety device according to claim 1, characterized in that the tunnel portions (A) are each provided with a traffic light, whereby in the event of a fire the fire sensor causes all traffic lights before the portion (A) with the fire to be switched to red and all traffic lights after the portion (A) with the fire to be switched to green."],["This invention relates to a safety device for fires in tunnels.","There has recently been a dramatic increase in the number of tunnel fires with human casualties.","Numerous elaborate measures have been proposed for diminishing the consequences, for example building rescue tunnels parallel to the existing tunnels.","The problem of the invention is to provide a simply constructed safety device for preventing the disastrous consequences of tunnel fires.","This is obtained according to the invention with the safety device characterized in claim 1.The subclaims render advantageous embodiments of the invention.","According to the invention, curtains that can be closed in the event of a fire are provided in the tunnel at intervals of for example 50 meters to 200 meters.","This permits the tunnel portion to be closed off when the fire sensor has detected a fire in said portion.","The curtain is made of incombustible material, for example a cloth of fibers of inorganic material.","The inventive safety device is thus characterized by its simplicity.","The fire sensor can be a smoke detector and/or heat sensor.","The curtains can be mounted on the tunnel ceiling in a rolled up or folded state.","However, the curtains are preferably drawn across the roadway or, in the case of a railroad or cable railway tunnel, across the tracks.","The curtains are for this purpose suspended on the tunnel wall in the drawn up, i.e.","folded, state, preferably in a joint or similar recess provided, for example milled, in the tunnel wall.","The recess or other space in which the drawn up curtains are located can be closable by flaps to protect the curtains from soiling.","Each curtain can consist of a plurality of parts, in particular two parts or halves, that in the drawn up or folded state are disposed on or in one or the other tunnel wall.","For closing, the two curtain parts are drawn in the direction of the opposite tunnel wall.","In the extended state the curtain parts can thus extend from one tunnel wall to the other, i.e.","form a double-walled curtain, but they can also extend only halfway across the tunnel, preferably so as to overlap in the middle of the tunnel.","Closing of the curtains is effected by a motor, for example an electric motor.","However, a pyrotechnic motor can also be provided for automatically closing the curtains for example.","The motor can also open the flaps protecting the curtains from soiling.","The curtains preferably consist in particular in the lower area of strips extending from above to below, i.e.","vertically, to permit them to be drawn over vehicles and similar objects in the tunnel.","The width of the strips can be for example 20 centimeters to 50 centimeters.","The strips are preferably formed so as to overlap in the extended state of the curtain.","To increase the protection, two curtain parts can also be provided on each side of the tunnel that form two also be provided on each side of the tunnel that form two curtain portions spaced one behind the other in the drawn up state of the curtain.","For drawing up, the curtains are guided on slide rails provided on or in the area of the tunnel ceiling.","If a plurality of parts are used for each curtain, multiple-track slide rails are preferably used.","In the event of a fire in one tunnel portion, the curtains of said portion are closed.","This contains the danger sector.","Persons located in the danger area thus have time to leave the danger area through the curtain.","The danger area is thus limited by the curtains to an easily surveyed sector.","Outside said area the smoke gases that have developed constitute a substantially lower danger for the individual persons.","To suck off the smoke gases arising in the event of a fire, each tunnel portion has at least one device for sucking off vitiated air.","The vitiated air suction device can be formed for example by an opening in the tunnel ceiling that leads to the vitiated air duct of the tunnel.","Preferably, the connection from each tunnel portion to the vitiated air duct is closable by a flap or similar shut-off device.","In the event of a fire the fire sensor can thus open the flap of the vitiated air suction device in the portion with the fire, while all or at least some of the flaps of the vitiated air suction devices of the other tunnel portions can be closed by the fire sensor.","The smoke gases formed can thus be selectively sucked off.","The closing of the flaps of the other tunnel portions causes the total suction effect to be concentrated on the portion with the fire.","The curtains are formed so that, in the closed state, they close off the upper area of the tunnel more tightly than the lower tunnel area.","This ensures that fresh air sucked into the portion with the fire by the vitiated air suction device flows into the lower area of the curtains, so that any injured persons lying on the tunnel floor have more chance of escaping the smoke gases.","It is known that 80 percent of tunnel victims die from smoke gases and not from burns.","Since the individual curtains are for example 100 meters apart, air is not sucked in strongly enough to cause additional kindling of the fire.","The inventive safety device guarantees that rescue vehicles and fire trucks can penetrate from outside to the portion with the fire without being exposed to appreciable smoke development or heat, which substantially facilitates rescue measures as well as extinguishing and salvage operations.","Further, the tunnel portions can each be provided with a traffic light, whereby in the event of a fire the fire sensor switches all traffic lights located before the portion with the fire in the driving direction to red, and all traffic lights located after the portion with the fire to green.","The traffic lights switched red cause the total following traffic to be not only stopped but at the same time broken up, since the individual cars or groups of cars stop at intervals, e.g.","of 100 meters, corresponding to the portions.","The broken up following traffic permits operational units to penetrate to the scene of the accident more easily.","This breaking up also permits vehicles to turn around more easily to leave the tunnel in the opposite direction.","The traffic lights switched green permit those vehicles to move away that are located after the portion with the fire in the driving direction.","Further, a loudspeaker or intercom can be mounted in each portion for guiding persons in the right direction.","Further, each portion can be provided with a video camera so that the circumstances of the accident can be immediately detected and the corresponding rescue measures taken.","To prevent false trips, a plurality of fire sensors are preferably provided in each portion.","Thus, the curtains are only closed before and after each portion in the driving direction if for example at least two fire sensors detect a fire.","Also, each tunnel portion can be provided with an impact switch for closing the curtains.","The actuation of the individual curtains, the individual sensors, the actuating devices for the flaps on the vitiated air suction device and the dust flaps for the drawn up curtains as well as the video cameras can be checked regularly by a central computer.","The inventive safety device permits tunnels to be retrofitted at reasonable cost.","In the event of a fire the relevant tunnel portion can be closed off within seconds.","Persons located in the danger area can flee through the preferably strip-shaped curtain and find safety.","Persons in the tunnel, outside the portion with the fire, are protected from smoke and heat even if they remain seated in their vehicles.","Smoke gases are sucked off from the source of the fire from above so that any injured persons on the tunnel floor are spared from smoke gases for a certain time.","Operational units can immediately penetrate to the danger area and begin rescue since the heat and source of the fire can be kept under control in the relevant portion.","Also, operational units can penetrate into the portion with the fire only to rescue people and then immediately leave it.","Thus, each person can move out of the danger area within seconds when passing the curtain.","Moreover, the power of the vitiated air suction device can be increased in the event of a fire.","In the following an embodiment of the inventive safety device for tunnel fires will be explained in more detail by way of example with reference to the drawing, in which: FIGS.","1 and 2 show a cross section through a tunnel with the curtain open and closed; and FIG.","3 shows a longitudinal section through a portion of the tunnel with closed curtains.","According to FIG.","3, the tunnel is divided into portions A that are separable from each other by fireproof curtains 1 mounted before and after portion A in the driving direction.","In each portion A fire sensors 2 are provided on the ceiling that close curtains 1 in the event of a fire in portion A.","According to FIG.","1, each curtain 1 consists of two curtain parts.","Drawn up curtain parts 3, 4 are disposed in recesses 5 on the left and right in the tunnel wall.","Recesses 5 are closable by flaps 6 to protect curtain parts 3, 4 from soiling.","Curtain parts 3, 4 are guided on slide rails 7 on tunnel ceiling 8.For closing, curtain parts 3, 4 are drawn across roadway 9 (FIG.","2) e.g.","with a motor not shown, for example an electric motor or similar automatically acting device not shown.","That is, curtain parts 3, 4 are extended in the direction of the other tunnel wall so that they overlap in the middle, as shown by the dash line in FIG.","2.Curtain parts 3, 4 consist of strips 11 that are 30 centimeters wide for example.","Curtain parts 3, 4 can also extend in the closed state across total roadway 9 so as to form a double-walled curtain comprising curtain parts 3 and 4, as to be seen in FIG.","3.Vitiated air duct 12 with ventilators and similar vitiated air suction devices not shown extends in the tunnel above tunnel ceiling 8.Vitiated air duct 12 is connected with portion A by suction openings 13.Suction openings 13 are closable with flaps 14.In the event of a fire, fire sensors 2 drive the motors for actuating curtains 1 of relevant portion A and the actuating devices for dirt flaps 6, causing flaps 6 to open and curtain parts 3, 4 to be drawn in the direction of the other tunnel wall for closing curtains 1.At the same time, sensors 2 cause flaps 14 to open for vitiated air suction in portion A with the fire, while the flaps are closed on the suction openings of the other portions, at least the portions adjacent the fire portion."]],"string":"[\n [\n \"Tunnel first safety device\",\n \"In order to increase the safety of tunnels when a fire occurs, the tunnel is divided into sections (A) which are then separated from each other by curtains (1).\",\n \"Each section (A) is provided with at least one fire sensor (2) which closes said curtains (1) in said section (A) if a fire breaks out.\"\n ],\n [\n \"1.A safety device for fires in tunnels, characterized in that the tunnel has portions (A) that are separable from each other by curtains (1), each portion (A) being provided with at least one fire sensor (2) that causes the curtains (1) of the portion (A) to be closed in the event of a fire.\",\n \"2.A safety device according to claim 1, characterized in that the curtains (1) are closed by a motor that is driven by the fire sensor.\",\n \"(2) 3.A safety device according to claim 1, characterized in that the curtains (1) are drawn across the roadway (9) for closing.\",\n \"4.A safety device according to claim 3, characterized in that the drawn up curtains (1) are disposed on the tunnel wall.\",\n \"5.A safety device according to claim 4, characterized in that the drawn up curtains (1) are disposed in a recess (5) in the tunnel wall.\",\n \"6.A safety device according to claim 1, characterized in that the drawn up curtains (1) are disposed in spaces closable by curtain flaps (6), the curtain flaps (6) being opened in the event of a fire.\",\n \"7.A safety device according to claim 1, characterized in that the curtains (1) consist of strips (11) extending from above to below.\",\n \"8.A safety device according to claim 7, characterized in that the strips (11) overlap in the extended state of the curtains (1).\",\n \"9.A safety device according to claim 1, characterized in that the curtains are formed to be fireproof.\",\n \"10.A safety device according to claim 1, characterized in that each curtain (1) consists of at least two curtain parts (3, 4) on each tunnel side that are drawn in the direction of the other tunnel side for closing.\",\n \"11.A safety device according to claim 1, characterized in that each curtain consists in the closed state of two portions (3, 4) spaced one behind the other.\",\n \"12.A safety device according to claim 1, characterized in that the curtains are guided on slide rails (7) provided in the area of the tunnel ceiling (8).\",\n \"13.A safety device according to claim 1, characterized in that the curtains (1) are formed in such a way as to close off the upper tunnel area more tightly than the lower tunnel area.\",\n \"14.A safety device according to claim 1, characterized in that each tunnel portion (A) is provided with at least one device for vitiated air suction.\",\n \"15.A safety device according to claim 14, characterized in that the vitiated air suction device has a connection to the vitiated air duct (12) of the tunnel, said connection being closable with a shut-off device (14).\",\n \"16.A safety device according to claim 15, characterized in that in the event of a fire the shut-off device (14) of the vitiated air device in the portion (A) with the fire is opened by the fire sensor (2) and the shut-off devices of the vitiated air suction devices of the other portions are at least partly closed.\",\n \"17.A safety device according to claim 1, characterized in that the tunnel portions (A) are each provided with a traffic light, whereby in the event of a fire the fire sensor causes all traffic lights before the portion (A) with the fire to be switched to red and all traffic lights after the portion (A) with the fire to be switched to green.\"\n ],\n [\n \"This invention relates to a safety device for fires in tunnels.\",\n \"There has recently been a dramatic increase in the number of tunnel fires with human casualties.\",\n \"Numerous elaborate measures have been proposed for diminishing the consequences, for example building rescue tunnels parallel to the existing tunnels.\",\n \"The problem of the invention is to provide a simply constructed safety device for preventing the disastrous consequences of tunnel fires.\",\n \"This is obtained according to the invention with the safety device characterized in claim 1.The subclaims render advantageous embodiments of the invention.\",\n \"According to the invention, curtains that can be closed in the event of a fire are provided in the tunnel at intervals of for example 50 meters to 200 meters.\",\n \"This permits the tunnel portion to be closed off when the fire sensor has detected a fire in said portion.\",\n \"The curtain is made of incombustible material, for example a cloth of fibers of inorganic material.\",\n \"The inventive safety device is thus characterized by its simplicity.\",\n \"The fire sensor can be a smoke detector and/or heat sensor.\",\n \"The curtains can be mounted on the tunnel ceiling in a rolled up or folded state.\",\n \"However, the curtains are preferably drawn across the roadway or, in the case of a railroad or cable railway tunnel, across the tracks.\",\n \"The curtains are for this purpose suspended on the tunnel wall in the drawn up, i.e.\",\n \"folded, state, preferably in a joint or similar recess provided, for example milled, in the tunnel wall.\",\n \"The recess or other space in which the drawn up curtains are located can be closable by flaps to protect the curtains from soiling.\",\n \"Each curtain can consist of a plurality of parts, in particular two parts or halves, that in the drawn up or folded state are disposed on or in one or the other tunnel wall.\",\n \"For closing, the two curtain parts are drawn in the direction of the opposite tunnel wall.\",\n \"In the extended state the curtain parts can thus extend from one tunnel wall to the other, i.e.\",\n \"form a double-walled curtain, but they can also extend only halfway across the tunnel, preferably so as to overlap in the middle of the tunnel.\",\n \"Closing of the curtains is effected by a motor, for example an electric motor.\",\n \"However, a pyrotechnic motor can also be provided for automatically closing the curtains for example.\",\n \"The motor can also open the flaps protecting the curtains from soiling.\",\n \"The curtains preferably consist in particular in the lower area of strips extending from above to below, i.e.\",\n \"vertically, to permit them to be drawn over vehicles and similar objects in the tunnel.\",\n \"The width of the strips can be for example 20 centimeters to 50 centimeters.\",\n \"The strips are preferably formed so as to overlap in the extended state of the curtain.\",\n \"To increase the protection, two curtain parts can also be provided on each side of the tunnel that form two also be provided on each side of the tunnel that form two curtain portions spaced one behind the other in the drawn up state of the curtain.\",\n \"For drawing up, the curtains are guided on slide rails provided on or in the area of the tunnel ceiling.\",\n \"If a plurality of parts are used for each curtain, multiple-track slide rails are preferably used.\",\n \"In the event of a fire in one tunnel portion, the curtains of said portion are closed.\",\n \"This contains the danger sector.\",\n \"Persons located in the danger area thus have time to leave the danger area through the curtain.\",\n \"The danger area is thus limited by the curtains to an easily surveyed sector.\",\n \"Outside said area the smoke gases that have developed constitute a substantially lower danger for the individual persons.\",\n \"To suck off the smoke gases arising in the event of a fire, each tunnel portion has at least one device for sucking off vitiated air.\",\n \"The vitiated air suction device can be formed for example by an opening in the tunnel ceiling that leads to the vitiated air duct of the tunnel.\",\n \"Preferably, the connection from each tunnel portion to the vitiated air duct is closable by a flap or similar shut-off device.\",\n \"In the event of a fire the fire sensor can thus open the flap of the vitiated air suction device in the portion with the fire, while all or at least some of the flaps of the vitiated air suction devices of the other tunnel portions can be closed by the fire sensor.\",\n \"The smoke gases formed can thus be selectively sucked off.\",\n \"The closing of the flaps of the other tunnel portions causes the total suction effect to be concentrated on the portion with the fire.\",\n \"The curtains are formed so that, in the closed state, they close off the upper area of the tunnel more tightly than the lower tunnel area.\",\n \"This ensures that fresh air sucked into the portion with the fire by the vitiated air suction device flows into the lower area of the curtains, so that any injured persons lying on the tunnel floor have more chance of escaping the smoke gases.\",\n \"It is known that 80 percent of tunnel victims die from smoke gases and not from burns.\",\n \"Since the individual curtains are for example 100 meters apart, air is not sucked in strongly enough to cause additional kindling of the fire.\",\n \"The inventive safety device guarantees that rescue vehicles and fire trucks can penetrate from outside to the portion with the fire without being exposed to appreciable smoke development or heat, which substantially facilitates rescue measures as well as extinguishing and salvage operations.\",\n \"Further, the tunnel portions can each be provided with a traffic light, whereby in the event of a fire the fire sensor switches all traffic lights located before the portion with the fire in the driving direction to red, and all traffic lights located after the portion with the fire to green.\",\n \"The traffic lights switched red cause the total following traffic to be not only stopped but at the same time broken up, since the individual cars or groups of cars stop at intervals, e.g.\",\n \"of 100 meters, corresponding to the portions.\",\n \"The broken up following traffic permits operational units to penetrate to the scene of the accident more easily.\",\n \"This breaking up also permits vehicles to turn around more easily to leave the tunnel in the opposite direction.\",\n \"The traffic lights switched green permit those vehicles to move away that are located after the portion with the fire in the driving direction.\",\n \"Further, a loudspeaker or intercom can be mounted in each portion for guiding persons in the right direction.\",\n \"Further, each portion can be provided with a video camera so that the circumstances of the accident can be immediately detected and the corresponding rescue measures taken.\",\n \"To prevent false trips, a plurality of fire sensors are preferably provided in each portion.\",\n \"Thus, the curtains are only closed before and after each portion in the driving direction if for example at least two fire sensors detect a fire.\",\n \"Also, each tunnel portion can be provided with an impact switch for closing the curtains.\",\n \"The actuation of the individual curtains, the individual sensors, the actuating devices for the flaps on the vitiated air suction device and the dust flaps for the drawn up curtains as well as the video cameras can be checked regularly by a central computer.\",\n \"The inventive safety device permits tunnels to be retrofitted at reasonable cost.\",\n \"In the event of a fire the relevant tunnel portion can be closed off within seconds.\",\n \"Persons located in the danger area can flee through the preferably strip-shaped curtain and find safety.\",\n \"Persons in the tunnel, outside the portion with the fire, are protected from smoke and heat even if they remain seated in their vehicles.\",\n \"Smoke gases are sucked off from the source of the fire from above so that any injured persons on the tunnel floor are spared from smoke gases for a certain time.\",\n \"Operational units can immediately penetrate to the danger area and begin rescue since the heat and source of the fire can be kept under control in the relevant portion.\",\n \"Also, operational units can penetrate into the portion with the fire only to rescue people and then immediately leave it.\",\n \"Thus, each person can move out of the danger area within seconds when passing the curtain.\",\n \"Moreover, the power of the vitiated air suction device can be increased in the event of a fire.\",\n \"In the following an embodiment of the inventive safety device for tunnel fires will be explained in more detail by way of example with reference to the drawing, in which: FIGS.\",\n \"1 and 2 show a cross section through a tunnel with the curtain open and closed; and FIG.\",\n \"3 shows a longitudinal section through a portion of the tunnel with closed curtains.\",\n \"According to FIG.\",\n \"3, the tunnel is divided into portions A that are separable from each other by fireproof curtains 1 mounted before and after portion A in the driving direction.\",\n \"In each portion A fire sensors 2 are provided on the ceiling that close curtains 1 in the event of a fire in portion A.\",\n \"According to FIG.\",\n \"1, each curtain 1 consists of two curtain parts.\",\n \"Drawn up curtain parts 3, 4 are disposed in recesses 5 on the left and right in the tunnel wall.\",\n \"Recesses 5 are closable by flaps 6 to protect curtain parts 3, 4 from soiling.\",\n \"Curtain parts 3, 4 are guided on slide rails 7 on tunnel ceiling 8.For closing, curtain parts 3, 4 are drawn across roadway 9 (FIG.\",\n \"2) e.g.\",\n \"with a motor not shown, for example an electric motor or similar automatically acting device not shown.\",\n \"That is, curtain parts 3, 4 are extended in the direction of the other tunnel wall so that they overlap in the middle, as shown by the dash line in FIG.\",\n \"2.Curtain parts 3, 4 consist of strips 11 that are 30 centimeters wide for example.\",\n \"Curtain parts 3, 4 can also extend in the closed state across total roadway 9 so as to form a double-walled curtain comprising curtain parts 3 and 4, as to be seen in FIG.\",\n \"3.Vitiated air duct 12 with ventilators and similar vitiated air suction devices not shown extends in the tunnel above tunnel ceiling 8.Vitiated air duct 12 is connected with portion A by suction openings 13.Suction openings 13 are closable with flaps 14.In the event of a fire, fire sensors 2 drive the motors for actuating curtains 1 of relevant portion A and the actuating devices for dirt flaps 6, causing flaps 6 to open and curtain parts 3, 4 to be drawn in the direction of the other tunnel wall for closing curtains 1.At the same time, sensors 2 cause flaps 14 to open for vitiated air suction in portion A with the fire, while the flaps are closed on the suction openings of the other portions, at least the portions adjacent the fire portion.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476930"}}},{"rowIdx":986,"cells":{"text":{"kind":"list like","value":[["Capacitive dynamometer","The invention relates to a capacitive dynamometer comprising an elastic body for receiving a mechanical force to be measured, the body having a measuring cavity, which is deformed, when the mechanical force to be measured is applied to the elastic body, the deformations of the inner wall of the cavity being picked up in the form of changes of capacity by means of capacitor electrodes co-operating with the inner wall.","The invention is characterized in that said elastic body includes a flexible tube having a length which is substantially larger than the height of said tube, said tube having solid ends and measuring means, adapted for measuring a displacement between said solid ends of said flexible tube, utilizing a cavity of said flexible tube adapted to performing or transferring the displacement measurement."],["1.A measuring device for measuring a mechanical force, said measuring device including an elastic body for receiving the mechanical force to be measured, said elastic body having a measuring cavity adapted for being deformed when the mechanical force to be measured is applied to the elastic body, said device including measuring means for measuring the deformation of said measuring cavity, characterized in that said elastic body includes a flexible tube having a length which is substantially larger than the height of said tube, said tube having solid ends, and said measuring means, adapted for measuring a displacement between said solid ends of said flexible tube, utilizing a cavity of said flexible tube adapted to performing or transferring the displacement measurement.","2.A measuring device according to claim 1, characterized in that said cavity of said flexible tube forms said measuring cavity.","3.A measuring device according to claim 1 or 2, characterized in that one or more of said solid ends include the measuring cavity.","4.A measuring device according to one or more of claims 1-3, characterized by including an extension of at least one said solid ends, said one or more extensions include the measuring cavity.","5.A measuring device according to one or more of claims 1-4, characterized in that said measuring means including an elongated electrode carrier mounted in said measuring cavity 6.A measuring device according to one or more of claims 1-5, characterized in that said measuring means including one or more measuring electrodes adapted for measuring a displacement between said electrode carrier and an inner wall of said cavity.","7.A measuring device according to one or more of claims 1-6, characterized by including supporting means adapted for supporting said electrode carrier in said measuring device, said supporting means including a first supporting member being located in the flexible tube.","8.A measuring device according to one or more of claims 1-7, characterized by including supporting means adapted for supporting said electrode carrier in said measuring device, said supporting means including a second supporting member being located in one or more of the solid ends.","9.A measuring device according to one or more of claims 1-8, characterized by including supporting means adapted for supporting said electrode carrier in said measuring device, said supporting means including a third supporting member being located in one or more of the extensions of the solid ends.","10.A measuring device according to one or more of claims 1-9, characterized in that one of said first, second and third supporting means forms a fixed support of said electrode carrier in said measuring device.","11.A measuring device according to one or more of the preceding claims, characterized in that said measuring means include capacitive and/or inductive electrodes.","12.A measuring device according to one or more of the preceding claims, characterized in that said one or more extensions of the said preferably solid ends of the flexible tube, include a solid cover to engage with said extension.","13.A measuring device according to one or more of the preceding claims, characterized in that said preferably solid ends of the flexible tube are tied together through beams, flexible mainly in the direction of the force to be measured."],["The invention relates to a capacitive dynamometer comprising an elastic body for receiving a mechanical force to be measured, the body having a measuring cavity, which is deformed, when the mechanical force to be measured is applied to the elastic body, the deformations of the inner wall of the cavity being picked up in the form of changes of capacity by means of capacitor electrodes co-operating with the inner wall.","From EP A 0.108.894 a torsion measuring device is known where capacitor electrodes are arranged in an elastic tube to detect the relative angular displacement of the ends of an elastic tube when a torsion is applied to the measuring device.","This measuring device is specifically designed to be insensitive to bending forces.","From U.S. Pat.","No.","4,175,428 a capacitive dynamometer of the kind referred to is known comprising an elastic body with a measuring cavity, which is deformed when a mechanical force to be measured is applied to the elastic body.","In this known dynamometer the cavity has to be of appreciable dimensions in order for the measured force to produce the dimensional changes that are necessary for the capacitance of the capacitor electrodes to change enough to be measured accurately.","As the elastic body is normally of steel and the electrode carrier is normally fabricated in a ceramic material with thick-film electrodes, the necessary large dimensions in connection with a rather high difference in coefficient of thermal expansion between steel and ceramics gives problems with measuring accuracy by varying temperature of the dynamometer.","Furthermore when the cavity is cylindrical with the measured force changing the cross section the whole length of the cavity, the necessary closure of the ends of the cavity against the environment has to be elastic and rather flexible in the form of a membrane or bellow in order not to influence the measurement.","It is the object of the invention to provide an improved capacitive dynamometer of the type initially mentioned.","According to the invention this object is achieved by a measuring device of the abovementioned type characterized in that said elastic body includes a flexible tube having a length which is substantially larger than the height of said tube, said tube having solid ends and measuring means, adapted for measuring a displacement between said solid ends of said flexible tube, utilizing a cavity of said flexible tube adapted to performing or transferring the displacement measurement.","Measuring cavities in the flexible tube and or in the solid ends or in extensions of the solid ends includes preferably an elongated electrode carrier, mounted in the measuring cavity, with one or more electrodes, which in connection with electrodes on the inner wall of the measuring cavity, forms measuring capacitances.","The preferably solid ends of the flexible tube are preferably tied together through beams, flexible mainly in the direction of the force to be measured, which forces the flexible tube to be deformed in a S shape when the measured force is applied to the dynamometer and which also makes the dynamometer substantially insensitive to torsion and to forces from all other directions than the direction of the measurement.","By this design according to the invention a range of advantages are obtained.","The design of the flexible elastic tube with a relatively low height gives, in combination with a certain length, a high flexibility with large displacements of the inner wall of the measuring cavity, and as these displacements are substantially at a right angle to the surface of the electrodes and substantially parallel to the direction of the measured force the important advantage is obtained that a very high proportion of the deflection of the measuring device is transformed in a displacement to be measured by the measuring capacitances.","An important advantage of the invention is that errors from thermal expansion due to differences in coefficients of thermal expansion of the elastic body and the electrode carrier are reduced because of the relatively small dimensions of the measuring cavity in the direction of displacements from the measured force, while at the same time a rather large difference of the thermal expansions of the electrode carrier and the flexible tube, because of their appreciable length, does not give measuring errors as this expansion takes place at a right angle to the direction of displacements used in the measurement.","The mounting of the electrode carrier in the measuring cavity is according to the invention performed by at least one support along the length of the electrode carrier.","The mounting of the electrode carrier in the measuring cavity is advantageously and according to the invention done by supports at both ends or at some distance from both ends of the flexible tube, whereby a high mechanical stability is obtained.","When the electrodes are positioned on the electrode carrier at positions opposite the inner wall of the flexible tube then, because the flexible tube is deformed in a S shape, the relative displacements are differential and changing sign along the length of the electrode carrier.","When the supports for the electrode carrier are designed to permit an angular movement of the electrode carrier relative to the solid ends, the electrode carrier is not bent and this way the differential displacements of the tube along its length and relative to the electrode carrier are according to the invention measured by placing electrodes acting as differential capacitors along the length of the electrode carrier at one or both sides.","By arranging electrodes at positions where the distance to the inner surface of the flexible tube is increasing and also where it is decreasing at maximum rate, when the force is applied, it is possible to obtain a high measuring signal which rejects the errors from parallel shifts of the electrode carrier and when additionally electrodes are placed where there is substantially no deformation it is also possible to get a rejection of errors from angular shifts of the electrode carrier in the cavity.","When the supports for the electrode carrier for stability reasons are designed to permit only a limited angular movement of the electrode carrier relative to the solid ends, the electrode carrier is partly bent and this way the differential displacements of the tube along its length and relative to the electrode carrier are different compared to the situation when the electrode carriers permit free angular movements.","When in another embodiment of the invention, the electrode carrier is mounted fixed at one end of the measuring cavity the measurement is performed by the changes of the capacitances formed between electrodes on the inner wall of the measuring cavity and electrodes positioned at the electrode carrier an appreciable distance from the mounting end of the electrode carrier.","A differential capacitor may preferably be implemented by corresponding electrodes on both the upper and the lower side of the electrode carrier.","In this embodiment the displacement of the inner wall of the cavity, relative to the measuring electrodes is large, which results in a high sensitivity, but errors resulting from angular movements of the electrode carrier at the mounting end may also be high.","The errors due to angular movements of the electrode carrier may however be compensated by electrodes placed near the fixed end of the electrode carrier, where the relative displacement from the force to be measured is lowest, in connection with calculations performed by the capacitance measuring circuit.","In a preferred embodiment of the invention the electrode carrier is formed by using a part of the material of the elastic body as the material for the electrode carrier, and it is preferably formed by the same machining process which forms the cavity of the elastic tube and it is preferably done in such a way that some of this material is left untouched to perform the function of supporting the electrode carrier in the cavity.","By this embodiment of the invention the stability of the electrode carrier is the highest possible.","Extensions of the preferably solid ends of the flexible tube gives the possibility to close the measuring cavity with solid covers which may be mounted at the ends without interfering with the deformations of the flexible tube.","The invention will now be described in further detail with reference to the drawing in which FIG.","1 illustrates an embodiment of a capacitive dynamometer according to the invention with the electrode carrier supported at both ends, FIG.","2 is a cross section at A-A of the capacitive dynamometer on FIG.","1, FIG.","3 is a top and side view of an electrode carrier with sets of measuring and compensation electrodes on both sides, FIG.","4 is the deflection, caused by the force to be measured, of the inner wall of the flexible tube relative to the fixed solid end of the flexible tube, FIG.","5 is the deflection of the electrode carrier supported at both ends in the cavity, and FIG.","6 is the difference between the deflection in FIG.","4 and the deflection in FIG.","5 i.e.","the displacement of the inner wall of the flexible tube relative to the electrode carrier.","FIG.","7 is a preferred embodiment of a capacitive dynamometer according to the invention with the electrode carrier supported at both ends, and with one of the supports placed in the cavity of one of the extensions of the solid ends, at a distance from one end of the flexible tube.","FIG.","8 is another preferred embodiment of a capacitive dynamometer according to the invention with the electrode carrier supported at both ends, and with one of the supports placed in the solid end of the flexible tube at a distance from the flexing part.","FIG.","9 is a preferred embodiment of a capacitive dynamometer according to the invention with the electrode carrier mounted fixed at one end of the flexible tube.","FIG.","10 is another preferred embodiment of a capacitive dynamometer according to the invention with the electrode carrier mounted fixed at one end of the flexible tube, and with the electrode carrier machined from a part of the material of the elastic body and consisting a part of this body.","FIG.","11 is a cross section at A-A of FIG.","10.FIG.","12 is a cross section at B-B of FIG.","10.FIG.","13 is a cross section at C-C of FIG.","10.FIG.","14 is another preferred embodiment of a capacitive dynamometer according to the invention with the electrode carrier mounted fixed at one end of the flexible tube, and with the electrode carrier machined from a part of the material of the elastic body and consisting a part of this body, and with measuring electrodes mounted fixed in the measuring cavity and the electrode carrier acting as the moving grounded counter electrode.","FIG.","1 is a side view of a preferred embodiment of the invention, showing the elastic body 1 with a force to be measured P applied to one end and with the other end kept fixed at a right angle to P. The elastic part in the form of the flexible tube 2 has the preferably solid ends 8 and 9.The electrode carrier 3 with electrodes 4 and 10 is shown mounted in the measuring cavity.","The flexible beams 5, ties together the solid ends of the flexible tube, in order to prevent the solid end 9 making an angular movement, but instead forces it to be displaced parallel to the force P. The combination of the beams and the solid ends is necessary for the flexible tube to deform in the S shape which again is a prerequisite for the function of the capacitive dynamometer according to FIG.","1.The beams are flexible because they have thinner cross sections 21 in order to concentrate their deflection at the ends.","The O-rings 6 are shown as an example of supports which keeps the electrode carrier in a fixed position preferably, but not necessarily, in the middle of the cavity of the elastic tube, as shown in FIG.","2, but at the same time allows the ends of the electrode carrier to turn freely in relation to the solid ends.","A further advantage by the O-rings 6 is their ability to seal the cavity.","The preferably solid ends 8 and 9 have extensions, which may be closed by solid covers 19 and 20, without interfering with the deformation of the flexible tube.","A capacitance measuring circuit 39, which is shown connected to the signal cable 7 and to a number of measuring and compensating electrodes on the electrode carrier 3, is placed in the extension of the solid end 8.When a force P is applied to the elastic body 1 a displacement of the flexible tube 2 and the beams 5, proportional to the force P, and depending on the dimensions of foremost the flexible tube, will take place.","A calculation of the displacement of a flexible tube 2, with the height 6 mm, cavity height 4 mm, width 25 mm, length 60 mm and force 100 N, is shown in FIG.","4 where the deflection in mm is shown on the y axis and the distance in mm along the length of the flexible tube is shown on the x axis.","The tube is here mounted fixed at the left end and loaded at the right end.","The ends of the electrode carrier 3 will follow the ends of the flexible tube and because the ends of the electrode carrier may turn freely in their supports in the solid ends of the flexible tube the electrode carrier will not be bent and the displacement of the inner wall relative to the electrode carrier will be as shown in FIG.","6 for the lower side of the electrode carrier.","A similar figure for the upper side of the electrode carrier will show the same displacements, but with opposite signs.","With the measuring electrodes arranged, as shown in FIG.","3, on one or preferably on both sides of the electrode carrier, and with electrode 4 in a location with the minimum and electrode 10 in a location with the maximum distance when the dynamometer is loaded, a differential signal is obtained which is seen to be compensated for errors due to a parallel shift of the electrode carrier in the cavity.","When electrodes are mounted in the corresponding places on the lower side of the electrode carrier a differential signal may likewise be obtained but with reversed polarities.","The function of the compensating electrodes 11, 12, 13 and 14 is to compensate errors due to angular shifts and bending of the electrode carrier in the cavity, by suitable choice of positions and areas for these electrodes.","When the electrodes 11 and 13 for example are coupled together and coupled differentially in relation to the measuring electrode 4 it is possible to fully compensate angular movements of the electrode carrier caused by movements up or down of one of the supports.","When the measuring electrode 4, according to the invention, is placed in a position with maximum relative displacement of the inner wall and the compensating electrodes 11 and 13 are placed at positions with minimum displacements, a high resulting signal may be obtained.","Likewise, the measuring electrode 10, may be compensated by the electrodes 12 and 14 and if measuring electrodes are mounted on the lower side of the electrode carrier corresponding compensating electrodes may be utilized the same way.","In a preferred embodiment the electrode carrier may also be supported at the middle where according to FIG.","6 the relative displacement is zero.","In FIG.","7 another embodiment of the invention is shown, where the mounting of one end of the electrode carrier 3 is made at a distance from the flexible tube 2 at the support 27, which here is mounted at the extension of the solid end 8.When a force P is applied to the dynamometer at the left end and the other end is fixed, the left end of the electrode carrier 3, which is supported at both ends in a way where an angular movement is permitted, will substantially have the same displacement as the solid end 9 while the other end of the electrode carrier will substantially have no displacement.","When electrodes 25 and 26 are positioned on the electrode carrier near the right end of the flexible tube they are positioned where the differential change of distance to the inner wall of the flexible tube may be high as it is the displacement due to the force P multiplied by the ratio of the distance from the left support to the position of the electrodes 25 and 26 to the distance between the supports.","These changes of distance from the electrodes 25 and 26 to the inner wall of the measuring cavity are seen to be differential.","The electrodes 23 and 24 are mounted in positions on the electrode carrier where the change of distance to the inner wall of the flexible tube is small and they may therefore be used as reference capacitors for the compensation of effects due to changing temperatures etc.","In FIG.","7 the capacitance measuring circuit 39 is shown mounted on the electrode carrier, but it may according to the invention instead be mounted separately anywhere in one of the extensions of the solid ends.","In FIG.","8, another preferred embodiment of the invention with substantially the same design as the dynamometer according to FIG.","7, but with one end of the electrode carrier 3, mounted at the lengthened solid end 8 of the flexible tube 2, providing the advantage of a very stable mounting, directly in the measuring cavity.","Here the electrodes 25 and 26 again are placed in positions with large differential changes of distance to the inner wall of the measuring cavity when the dynamometer is measuring the force P. In this embodiment of the invention it is also possible to compensate errors from movements of the electrode carrier in the measuring cavity by sets of reference electrodes 23, 24 and 28 and 29.In FIG.","9 an embodiment of the invention is shown where the electrode carrier 3 is mounted at the solid end 8 of the flexible tube 2 by the dimensionally stable mounting bracket 18.The differentially coupled electrodes 22 and 15 measures the deflection of the flexible tube at or near the solid end 9, which has a rather large deflection as shown in FIG.","4.This embodiment of the invention is seen to be sensitive to angular movements of the electrode carrier relative to the solid end 8, but the compensating electrodes 16 and 17, placed in positions with low displacements from the force to be measured, will predominantly sense only this angular movement and may in connection with calculations performed by the capacitance measuring circuit compensate the errors from the angular movements.","An embodiment of the capacitive dynamometer according to FIG.","9, without the beams 5, will be sensitive to the method of application of the force P, but may be produced at a very low cost.","This embodiment of the invention is advantageous in applications with high demands on sensitivity, but with an environment with low vibrations and stable temperatures.","According to the invention the electrode carrier may, as shown in FIG.","9, be extended outside the measuring cavity with the capacitance measuring circuit 39 for the measurement of the capacitances mounted on the extension with the advantage that a complete measuring unit may simply be inserted into the cavity of the elastic body to obtain the lowest possible assembly costs.","The electrode carrier may advantageously be made of a dimensionally stable insulating material such as ceramic or glass-ceramic materials with electrodes deposited on the surface by for example thick- or thin film techniques or the electrode carrier may be made of metal or of a metal with an insulating layer as the basis for depositing the electrodes, but other methods of producing the electrodes may be used according to the invention.","As an example, for low-cost versions of the capacitive dynamometer, the electrode carrier may by printed circuit material where the electrodes are etched on the base material.","The electrode carrier may advantageously also be produced by sandwiching a stiffening layer of insulating or non insulating material between two electrode carriers with the measuring electrodes deposited on the outer side and the interconnections to the capacitance measuring circuit or the measuring circuit itself mounted on the inner side of the electrode carriers or between these.","In another embodiment of the invention the electrode carrier is made of a relatively thick material and here the interconnections may advantageously be placed on the side of the electrode carrier in order not to interfere with the measurement.","In FIG.","10 is shown an embodiment of the invention, with the electrode carrier 3, machined from the base material of the elastic body 1, with one end of the electrode carrier still attached to the base material, while the free end carries the electrodes 30 and 31 which measures the differential changes of distance to the inner wall of the measuring cavity when the dynamometer measures the force P. The preferred method of machining is spark erosion machining which gives a limited distortion of the material, which again gives a very high stability of the electrode carrier in the measuring cavity which here is a part of the solid end 9.The electrodes 30 and 31 may advantageously be mounted on insulators which are fixed in relation to the electrode carrier 3 in a way to permit the mounting of the reference electrodes 32 and 33.These electrodes will basically see no change of distance in the normal operation of the dynamometer, but may be connected to compensate displacements of the insulators carrying the measuring electrodes 30 and 31 in relation to the electrode carrier 3.The electrodes 30 and 33 may be connected together or they may simply constitute the upper and the lower surface of an electrode produced from an electrically conducting material, the same applies of course for electrodes 31 and 32.FIG.","11 is a cross section of the dynamometer at A-A showing the end of the electrode carrier 3 with the electrodes 30 and 31 positioned in the measuring cavity, machined in the solid end 9.FIG.","12 is a cross section at B-B of the dynamometer showing the electrode carrier 3 in the flexible tube 2 and the beams 5.FIG.","13 is a cross section at C-C, showing holes 33 and 34 drilled through the solid end 8 where the electrode carrier is fixed in the base material of the elastic body.","The function of the holes is to conduct the signals from the electrodes 30, 31 and 32, 33 to the capacitance measuring circuit 39.While the process of machining the electrode carrier from the base material of the dynamometer gives the highest stability it is according to the invention possible to mount a separately machined electrode carrier as shown in FIG.","9 in a number of ways, as for example by directly laser welding the electrode carrier 3 into the solid end 8, thereby obtaining the same function as in FIG.","10, but with less stability and at a lower cost.","An advantage of the dynamometer according to the invention and as shown in FIGS.","9 and 10 is that shock loads from the force P are only sensed by the robust elastic body 1 with the flexible tube 2 and the beams 5, because the electrode carrier 3 with the electrodes and the capacitance measuring circuit 12 are referenced to the fixed solid end of the dynamometer.","In FIG.","14 a dynamometer according to the invention is shown where the function of the electrode carrier 3 is to act as the grounded moving electrode while the electrodes 35 and 36, which are fixed in the measuring cavity, machined in the solid end 9, differentially measures the displacement of the electrode carrier 3.Reference electrodes 37 and 38 may compensate displacements, due to temperature changes, of the insulators of electrodes 35 and 36 relative to the measuring cavity in the same way as in the dynamometer according to FIG.","10.The electrodes 35 and 37 may be connected together or they may simply constitute the upper and the lower surface of an electrode produced from an electrically conducting material, the same applies of course for electrodes 36 and 38.This dynamometer according to the invention has the same advantages as the dynamometers according to FIGS.","9 and 10, but with a simpler connection from the electrodes to the measuring circuit 39.The electrode carrier 3 may be made stiff enough to place its frequency of resonance above the frequencies introduced by the force P. Due to the fact that preferred embodiments of the invention have been illustrated and described herein, it will be apparent to those skilled in the art that modifications and improvements may be made to forms herein specifically disclosed.","Accordingly, the present invention, which basically consists of two solid ends connected by a flexible tube, each solid end containing a part of a displacement measuring system, is not to be limited to the forms specifically disclosed.","For example the measuring electrodes measuring the displacements, due to the force to be measured, of one of the solid ends in relation to the other solid end, may in all embodiments of the dynamometer according to the invention be placed fixed either on the electrode carrier or fixed in the measuring cavity.","Correspondingly the counter electrode, which normally, but not necessarily is grounded, may either be placed in the measuring cavity or on the electrode carrier, or the electrode carrier itself could form the electrode.","Likewise the part which is displaced due to the force to be measured may be the measuring cavity or the electrode carrier.","As another example, insulated electrodes could be placed both on the electrode carrier and in the measuring cavity in a measuring device according to the invention.","Furthermore for example the cross section of the flexible tube may according to the invention be oval or round, square or rectangular or alternatively have a cavity with a cross section with rounded ends.","Also the disclosed capacitive method of measuring the changes of displacements may be substituted by other displacement measuring methods, for example eddy current measurements where the capacitive electrodes are simply substituted by small coils."]],"string":"[\n [\n \"Capacitive dynamometer\",\n \"The invention relates to a capacitive dynamometer comprising an elastic body for receiving a mechanical force to be measured, the body having a measuring cavity, which is deformed, when the mechanical force to be measured is applied to the elastic body, the deformations of the inner wall of the cavity being picked up in the form of changes of capacity by means of capacitor electrodes co-operating with the inner wall.\",\n \"The invention is characterized in that said elastic body includes a flexible tube having a length which is substantially larger than the height of said tube, said tube having solid ends and measuring means, adapted for measuring a displacement between said solid ends of said flexible tube, utilizing a cavity of said flexible tube adapted to performing or transferring the displacement measurement.\"\n ],\n [\n \"1.A measuring device for measuring a mechanical force, said measuring device including an elastic body for receiving the mechanical force to be measured, said elastic body having a measuring cavity adapted for being deformed when the mechanical force to be measured is applied to the elastic body, said device including measuring means for measuring the deformation of said measuring cavity, characterized in that said elastic body includes a flexible tube having a length which is substantially larger than the height of said tube, said tube having solid ends, and said measuring means, adapted for measuring a displacement between said solid ends of said flexible tube, utilizing a cavity of said flexible tube adapted to performing or transferring the displacement measurement.\",\n \"2.A measuring device according to claim 1, characterized in that said cavity of said flexible tube forms said measuring cavity.\",\n \"3.A measuring device according to claim 1 or 2, characterized in that one or more of said solid ends include the measuring cavity.\",\n \"4.A measuring device according to one or more of claims 1-3, characterized by including an extension of at least one said solid ends, said one or more extensions include the measuring cavity.\",\n \"5.A measuring device according to one or more of claims 1-4, characterized in that said measuring means including an elongated electrode carrier mounted in said measuring cavity 6.A measuring device according to one or more of claims 1-5, characterized in that said measuring means including one or more measuring electrodes adapted for measuring a displacement between said electrode carrier and an inner wall of said cavity.\",\n \"7.A measuring device according to one or more of claims 1-6, characterized by including supporting means adapted for supporting said electrode carrier in said measuring device, said supporting means including a first supporting member being located in the flexible tube.\",\n \"8.A measuring device according to one or more of claims 1-7, characterized by including supporting means adapted for supporting said electrode carrier in said measuring device, said supporting means including a second supporting member being located in one or more of the solid ends.\",\n \"9.A measuring device according to one or more of claims 1-8, characterized by including supporting means adapted for supporting said electrode carrier in said measuring device, said supporting means including a third supporting member being located in one or more of the extensions of the solid ends.\",\n \"10.A measuring device according to one or more of claims 1-9, characterized in that one of said first, second and third supporting means forms a fixed support of said electrode carrier in said measuring device.\",\n \"11.A measuring device according to one or more of the preceding claims, characterized in that said measuring means include capacitive and/or inductive electrodes.\",\n \"12.A measuring device according to one or more of the preceding claims, characterized in that said one or more extensions of the said preferably solid ends of the flexible tube, include a solid cover to engage with said extension.\",\n \"13.A measuring device according to one or more of the preceding claims, characterized in that said preferably solid ends of the flexible tube are tied together through beams, flexible mainly in the direction of the force to be measured.\"\n ],\n [\n \"The invention relates to a capacitive dynamometer comprising an elastic body for receiving a mechanical force to be measured, the body having a measuring cavity, which is deformed, when the mechanical force to be measured is applied to the elastic body, the deformations of the inner wall of the cavity being picked up in the form of changes of capacity by means of capacitor electrodes co-operating with the inner wall.\",\n \"From EP A 0.108.894 a torsion measuring device is known where capacitor electrodes are arranged in an elastic tube to detect the relative angular displacement of the ends of an elastic tube when a torsion is applied to the measuring device.\",\n \"This measuring device is specifically designed to be insensitive to bending forces.\",\n \"From U.S. Pat.\",\n \"No.\",\n \"4,175,428 a capacitive dynamometer of the kind referred to is known comprising an elastic body with a measuring cavity, which is deformed when a mechanical force to be measured is applied to the elastic body.\",\n \"In this known dynamometer the cavity has to be of appreciable dimensions in order for the measured force to produce the dimensional changes that are necessary for the capacitance of the capacitor electrodes to change enough to be measured accurately.\",\n \"As the elastic body is normally of steel and the electrode carrier is normally fabricated in a ceramic material with thick-film electrodes, the necessary large dimensions in connection with a rather high difference in coefficient of thermal expansion between steel and ceramics gives problems with measuring accuracy by varying temperature of the dynamometer.\",\n \"Furthermore when the cavity is cylindrical with the measured force changing the cross section the whole length of the cavity, the necessary closure of the ends of the cavity against the environment has to be elastic and rather flexible in the form of a membrane or bellow in order not to influence the measurement.\",\n \"It is the object of the invention to provide an improved capacitive dynamometer of the type initially mentioned.\",\n \"According to the invention this object is achieved by a measuring device of the abovementioned type characterized in that said elastic body includes a flexible tube having a length which is substantially larger than the height of said tube, said tube having solid ends and measuring means, adapted for measuring a displacement between said solid ends of said flexible tube, utilizing a cavity of said flexible tube adapted to performing or transferring the displacement measurement.\",\n \"Measuring cavities in the flexible tube and or in the solid ends or in extensions of the solid ends includes preferably an elongated electrode carrier, mounted in the measuring cavity, with one or more electrodes, which in connection with electrodes on the inner wall of the measuring cavity, forms measuring capacitances.\",\n \"The preferably solid ends of the flexible tube are preferably tied together through beams, flexible mainly in the direction of the force to be measured, which forces the flexible tube to be deformed in a S shape when the measured force is applied to the dynamometer and which also makes the dynamometer substantially insensitive to torsion and to forces from all other directions than the direction of the measurement.\",\n \"By this design according to the invention a range of advantages are obtained.\",\n \"The design of the flexible elastic tube with a relatively low height gives, in combination with a certain length, a high flexibility with large displacements of the inner wall of the measuring cavity, and as these displacements are substantially at a right angle to the surface of the electrodes and substantially parallel to the direction of the measured force the important advantage is obtained that a very high proportion of the deflection of the measuring device is transformed in a displacement to be measured by the measuring capacitances.\",\n \"An important advantage of the invention is that errors from thermal expansion due to differences in coefficients of thermal expansion of the elastic body and the electrode carrier are reduced because of the relatively small dimensions of the measuring cavity in the direction of displacements from the measured force, while at the same time a rather large difference of the thermal expansions of the electrode carrier and the flexible tube, because of their appreciable length, does not give measuring errors as this expansion takes place at a right angle to the direction of displacements used in the measurement.\",\n \"The mounting of the electrode carrier in the measuring cavity is according to the invention performed by at least one support along the length of the electrode carrier.\",\n \"The mounting of the electrode carrier in the measuring cavity is advantageously and according to the invention done by supports at both ends or at some distance from both ends of the flexible tube, whereby a high mechanical stability is obtained.\",\n \"When the electrodes are positioned on the electrode carrier at positions opposite the inner wall of the flexible tube then, because the flexible tube is deformed in a S shape, the relative displacements are differential and changing sign along the length of the electrode carrier.\",\n \"When the supports for the electrode carrier are designed to permit an angular movement of the electrode carrier relative to the solid ends, the electrode carrier is not bent and this way the differential displacements of the tube along its length and relative to the electrode carrier are according to the invention measured by placing electrodes acting as differential capacitors along the length of the electrode carrier at one or both sides.\",\n \"By arranging electrodes at positions where the distance to the inner surface of the flexible tube is increasing and also where it is decreasing at maximum rate, when the force is applied, it is possible to obtain a high measuring signal which rejects the errors from parallel shifts of the electrode carrier and when additionally electrodes are placed where there is substantially no deformation it is also possible to get a rejection of errors from angular shifts of the electrode carrier in the cavity.\",\n \"When the supports for the electrode carrier for stability reasons are designed to permit only a limited angular movement of the electrode carrier relative to the solid ends, the electrode carrier is partly bent and this way the differential displacements of the tube along its length and relative to the electrode carrier are different compared to the situation when the electrode carriers permit free angular movements.\",\n \"When in another embodiment of the invention, the electrode carrier is mounted fixed at one end of the measuring cavity the measurement is performed by the changes of the capacitances formed between electrodes on the inner wall of the measuring cavity and electrodes positioned at the electrode carrier an appreciable distance from the mounting end of the electrode carrier.\",\n \"A differential capacitor may preferably be implemented by corresponding electrodes on both the upper and the lower side of the electrode carrier.\",\n \"In this embodiment the displacement of the inner wall of the cavity, relative to the measuring electrodes is large, which results in a high sensitivity, but errors resulting from angular movements of the electrode carrier at the mounting end may also be high.\",\n \"The errors due to angular movements of the electrode carrier may however be compensated by electrodes placed near the fixed end of the electrode carrier, where the relative displacement from the force to be measured is lowest, in connection with calculations performed by the capacitance measuring circuit.\",\n \"In a preferred embodiment of the invention the electrode carrier is formed by using a part of the material of the elastic body as the material for the electrode carrier, and it is preferably formed by the same machining process which forms the cavity of the elastic tube and it is preferably done in such a way that some of this material is left untouched to perform the function of supporting the electrode carrier in the cavity.\",\n \"By this embodiment of the invention the stability of the electrode carrier is the highest possible.\",\n \"Extensions of the preferably solid ends of the flexible tube gives the possibility to close the measuring cavity with solid covers which may be mounted at the ends without interfering with the deformations of the flexible tube.\",\n \"The invention will now be described in further detail with reference to the drawing in which FIG.\",\n \"1 illustrates an embodiment of a capacitive dynamometer according to the invention with the electrode carrier supported at both ends, FIG.\",\n \"2 is a cross section at A-A of the capacitive dynamometer on FIG.\",\n \"1, FIG.\",\n \"3 is a top and side view of an electrode carrier with sets of measuring and compensation electrodes on both sides, FIG.\",\n \"4 is the deflection, caused by the force to be measured, of the inner wall of the flexible tube relative to the fixed solid end of the flexible tube, FIG.\",\n \"5 is the deflection of the electrode carrier supported at both ends in the cavity, and FIG.\",\n \"6 is the difference between the deflection in FIG.\",\n \"4 and the deflection in FIG.\",\n \"5 i.e.\",\n \"the displacement of the inner wall of the flexible tube relative to the electrode carrier.\",\n \"FIG.\",\n \"7 is a preferred embodiment of a capacitive dynamometer according to the invention with the electrode carrier supported at both ends, and with one of the supports placed in the cavity of one of the extensions of the solid ends, at a distance from one end of the flexible tube.\",\n \"FIG.\",\n \"8 is another preferred embodiment of a capacitive dynamometer according to the invention with the electrode carrier supported at both ends, and with one of the supports placed in the solid end of the flexible tube at a distance from the flexing part.\",\n \"FIG.\",\n \"9 is a preferred embodiment of a capacitive dynamometer according to the invention with the electrode carrier mounted fixed at one end of the flexible tube.\",\n \"FIG.\",\n \"10 is another preferred embodiment of a capacitive dynamometer according to the invention with the electrode carrier mounted fixed at one end of the flexible tube, and with the electrode carrier machined from a part of the material of the elastic body and consisting a part of this body.\",\n \"FIG.\",\n \"11 is a cross section at A-A of FIG.\",\n \"10.FIG.\",\n \"12 is a cross section at B-B of FIG.\",\n \"10.FIG.\",\n \"13 is a cross section at C-C of FIG.\",\n \"10.FIG.\",\n \"14 is another preferred embodiment of a capacitive dynamometer according to the invention with the electrode carrier mounted fixed at one end of the flexible tube, and with the electrode carrier machined from a part of the material of the elastic body and consisting a part of this body, and with measuring electrodes mounted fixed in the measuring cavity and the electrode carrier acting as the moving grounded counter electrode.\",\n \"FIG.\",\n \"1 is a side view of a preferred embodiment of the invention, showing the elastic body 1 with a force to be measured P applied to one end and with the other end kept fixed at a right angle to P. The elastic part in the form of the flexible tube 2 has the preferably solid ends 8 and 9.The electrode carrier 3 with electrodes 4 and 10 is shown mounted in the measuring cavity.\",\n \"The flexible beams 5, ties together the solid ends of the flexible tube, in order to prevent the solid end 9 making an angular movement, but instead forces it to be displaced parallel to the force P. The combination of the beams and the solid ends is necessary for the flexible tube to deform in the S shape which again is a prerequisite for the function of the capacitive dynamometer according to FIG.\",\n \"1.The beams are flexible because they have thinner cross sections 21 in order to concentrate their deflection at the ends.\",\n \"The O-rings 6 are shown as an example of supports which keeps the electrode carrier in a fixed position preferably, but not necessarily, in the middle of the cavity of the elastic tube, as shown in FIG.\",\n \"2, but at the same time allows the ends of the electrode carrier to turn freely in relation to the solid ends.\",\n \"A further advantage by the O-rings 6 is their ability to seal the cavity.\",\n \"The preferably solid ends 8 and 9 have extensions, which may be closed by solid covers 19 and 20, without interfering with the deformation of the flexible tube.\",\n \"A capacitance measuring circuit 39, which is shown connected to the signal cable 7 and to a number of measuring and compensating electrodes on the electrode carrier 3, is placed in the extension of the solid end 8.When a force P is applied to the elastic body 1 a displacement of the flexible tube 2 and the beams 5, proportional to the force P, and depending on the dimensions of foremost the flexible tube, will take place.\",\n \"A calculation of the displacement of a flexible tube 2, with the height 6 mm, cavity height 4 mm, width 25 mm, length 60 mm and force 100 N, is shown in FIG.\",\n \"4 where the deflection in mm is shown on the y axis and the distance in mm along the length of the flexible tube is shown on the x axis.\",\n \"The tube is here mounted fixed at the left end and loaded at the right end.\",\n \"The ends of the electrode carrier 3 will follow the ends of the flexible tube and because the ends of the electrode carrier may turn freely in their supports in the solid ends of the flexible tube the electrode carrier will not be bent and the displacement of the inner wall relative to the electrode carrier will be as shown in FIG.\",\n \"6 for the lower side of the electrode carrier.\",\n \"A similar figure for the upper side of the electrode carrier will show the same displacements, but with opposite signs.\",\n \"With the measuring electrodes arranged, as shown in FIG.\",\n \"3, on one or preferably on both sides of the electrode carrier, and with electrode 4 in a location with the minimum and electrode 10 in a location with the maximum distance when the dynamometer is loaded, a differential signal is obtained which is seen to be compensated for errors due to a parallel shift of the electrode carrier in the cavity.\",\n \"When electrodes are mounted in the corresponding places on the lower side of the electrode carrier a differential signal may likewise be obtained but with reversed polarities.\",\n \"The function of the compensating electrodes 11, 12, 13 and 14 is to compensate errors due to angular shifts and bending of the electrode carrier in the cavity, by suitable choice of positions and areas for these electrodes.\",\n \"When the electrodes 11 and 13 for example are coupled together and coupled differentially in relation to the measuring electrode 4 it is possible to fully compensate angular movements of the electrode carrier caused by movements up or down of one of the supports.\",\n \"When the measuring electrode 4, according to the invention, is placed in a position with maximum relative displacement of the inner wall and the compensating electrodes 11 and 13 are placed at positions with minimum displacements, a high resulting signal may be obtained.\",\n \"Likewise, the measuring electrode 10, may be compensated by the electrodes 12 and 14 and if measuring electrodes are mounted on the lower side of the electrode carrier corresponding compensating electrodes may be utilized the same way.\",\n \"In a preferred embodiment the electrode carrier may also be supported at the middle where according to FIG.\",\n \"6 the relative displacement is zero.\",\n \"In FIG.\",\n \"7 another embodiment of the invention is shown, where the mounting of one end of the electrode carrier 3 is made at a distance from the flexible tube 2 at the support 27, which here is mounted at the extension of the solid end 8.When a force P is applied to the dynamometer at the left end and the other end is fixed, the left end of the electrode carrier 3, which is supported at both ends in a way where an angular movement is permitted, will substantially have the same displacement as the solid end 9 while the other end of the electrode carrier will substantially have no displacement.\",\n \"When electrodes 25 and 26 are positioned on the electrode carrier near the right end of the flexible tube they are positioned where the differential change of distance to the inner wall of the flexible tube may be high as it is the displacement due to the force P multiplied by the ratio of the distance from the left support to the position of the electrodes 25 and 26 to the distance between the supports.\",\n \"These changes of distance from the electrodes 25 and 26 to the inner wall of the measuring cavity are seen to be differential.\",\n \"The electrodes 23 and 24 are mounted in positions on the electrode carrier where the change of distance to the inner wall of the flexible tube is small and they may therefore be used as reference capacitors for the compensation of effects due to changing temperatures etc.\",\n \"In FIG.\",\n \"7 the capacitance measuring circuit 39 is shown mounted on the electrode carrier, but it may according to the invention instead be mounted separately anywhere in one of the extensions of the solid ends.\",\n \"In FIG.\",\n \"8, another preferred embodiment of the invention with substantially the same design as the dynamometer according to FIG.\",\n \"7, but with one end of the electrode carrier 3, mounted at the lengthened solid end 8 of the flexible tube 2, providing the advantage of a very stable mounting, directly in the measuring cavity.\",\n \"Here the electrodes 25 and 26 again are placed in positions with large differential changes of distance to the inner wall of the measuring cavity when the dynamometer is measuring the force P. In this embodiment of the invention it is also possible to compensate errors from movements of the electrode carrier in the measuring cavity by sets of reference electrodes 23, 24 and 28 and 29.In FIG.\",\n \"9 an embodiment of the invention is shown where the electrode carrier 3 is mounted at the solid end 8 of the flexible tube 2 by the dimensionally stable mounting bracket 18.The differentially coupled electrodes 22 and 15 measures the deflection of the flexible tube at or near the solid end 9, which has a rather large deflection as shown in FIG.\",\n \"4.This embodiment of the invention is seen to be sensitive to angular movements of the electrode carrier relative to the solid end 8, but the compensating electrodes 16 and 17, placed in positions with low displacements from the force to be measured, will predominantly sense only this angular movement and may in connection with calculations performed by the capacitance measuring circuit compensate the errors from the angular movements.\",\n \"An embodiment of the capacitive dynamometer according to FIG.\",\n \"9, without the beams 5, will be sensitive to the method of application of the force P, but may be produced at a very low cost.\",\n \"This embodiment of the invention is advantageous in applications with high demands on sensitivity, but with an environment with low vibrations and stable temperatures.\",\n \"According to the invention the electrode carrier may, as shown in FIG.\",\n \"9, be extended outside the measuring cavity with the capacitance measuring circuit 39 for the measurement of the capacitances mounted on the extension with the advantage that a complete measuring unit may simply be inserted into the cavity of the elastic body to obtain the lowest possible assembly costs.\",\n \"The electrode carrier may advantageously be made of a dimensionally stable insulating material such as ceramic or glass-ceramic materials with electrodes deposited on the surface by for example thick- or thin film techniques or the electrode carrier may be made of metal or of a metal with an insulating layer as the basis for depositing the electrodes, but other methods of producing the electrodes may be used according to the invention.\",\n \"As an example, for low-cost versions of the capacitive dynamometer, the electrode carrier may by printed circuit material where the electrodes are etched on the base material.\",\n \"The electrode carrier may advantageously also be produced by sandwiching a stiffening layer of insulating or non insulating material between two electrode carriers with the measuring electrodes deposited on the outer side and the interconnections to the capacitance measuring circuit or the measuring circuit itself mounted on the inner side of the electrode carriers or between these.\",\n \"In another embodiment of the invention the electrode carrier is made of a relatively thick material and here the interconnections may advantageously be placed on the side of the electrode carrier in order not to interfere with the measurement.\",\n \"In FIG.\",\n \"10 is shown an embodiment of the invention, with the electrode carrier 3, machined from the base material of the elastic body 1, with one end of the electrode carrier still attached to the base material, while the free end carries the electrodes 30 and 31 which measures the differential changes of distance to the inner wall of the measuring cavity when the dynamometer measures the force P. The preferred method of machining is spark erosion machining which gives a limited distortion of the material, which again gives a very high stability of the electrode carrier in the measuring cavity which here is a part of the solid end 9.The electrodes 30 and 31 may advantageously be mounted on insulators which are fixed in relation to the electrode carrier 3 in a way to permit the mounting of the reference electrodes 32 and 33.These electrodes will basically see no change of distance in the normal operation of the dynamometer, but may be connected to compensate displacements of the insulators carrying the measuring electrodes 30 and 31 in relation to the electrode carrier 3.The electrodes 30 and 33 may be connected together or they may simply constitute the upper and the lower surface of an electrode produced from an electrically conducting material, the same applies of course for electrodes 31 and 32.FIG.\",\n \"11 is a cross section of the dynamometer at A-A showing the end of the electrode carrier 3 with the electrodes 30 and 31 positioned in the measuring cavity, machined in the solid end 9.FIG.\",\n \"12 is a cross section at B-B of the dynamometer showing the electrode carrier 3 in the flexible tube 2 and the beams 5.FIG.\",\n \"13 is a cross section at C-C, showing holes 33 and 34 drilled through the solid end 8 where the electrode carrier is fixed in the base material of the elastic body.\",\n \"The function of the holes is to conduct the signals from the electrodes 30, 31 and 32, 33 to the capacitance measuring circuit 39.While the process of machining the electrode carrier from the base material of the dynamometer gives the highest stability it is according to the invention possible to mount a separately machined electrode carrier as shown in FIG.\",\n \"9 in a number of ways, as for example by directly laser welding the electrode carrier 3 into the solid end 8, thereby obtaining the same function as in FIG.\",\n \"10, but with less stability and at a lower cost.\",\n \"An advantage of the dynamometer according to the invention and as shown in FIGS.\",\n \"9 and 10 is that shock loads from the force P are only sensed by the robust elastic body 1 with the flexible tube 2 and the beams 5, because the electrode carrier 3 with the electrodes and the capacitance measuring circuit 12 are referenced to the fixed solid end of the dynamometer.\",\n \"In FIG.\",\n \"14 a dynamometer according to the invention is shown where the function of the electrode carrier 3 is to act as the grounded moving electrode while the electrodes 35 and 36, which are fixed in the measuring cavity, machined in the solid end 9, differentially measures the displacement of the electrode carrier 3.Reference electrodes 37 and 38 may compensate displacements, due to temperature changes, of the insulators of electrodes 35 and 36 relative to the measuring cavity in the same way as in the dynamometer according to FIG.\",\n \"10.The electrodes 35 and 37 may be connected together or they may simply constitute the upper and the lower surface of an electrode produced from an electrically conducting material, the same applies of course for electrodes 36 and 38.This dynamometer according to the invention has the same advantages as the dynamometers according to FIGS.\",\n \"9 and 10, but with a simpler connection from the electrodes to the measuring circuit 39.The electrode carrier 3 may be made stiff enough to place its frequency of resonance above the frequencies introduced by the force P. Due to the fact that preferred embodiments of the invention have been illustrated and described herein, it will be apparent to those skilled in the art that modifications and improvements may be made to forms herein specifically disclosed.\",\n \"Accordingly, the present invention, which basically consists of two solid ends connected by a flexible tube, each solid end containing a part of a displacement measuring system, is not to be limited to the forms specifically disclosed.\",\n \"For example the measuring electrodes measuring the displacements, due to the force to be measured, of one of the solid ends in relation to the other solid end, may in all embodiments of the dynamometer according to the invention be placed fixed either on the electrode carrier or fixed in the measuring cavity.\",\n \"Correspondingly the counter electrode, which normally, but not necessarily is grounded, may either be placed in the measuring cavity or on the electrode carrier, or the electrode carrier itself could form the electrode.\",\n \"Likewise the part which is displaced due to the force to be measured may be the measuring cavity or the electrode carrier.\",\n \"As another example, insulated electrodes could be placed both on the electrode carrier and in the measuring cavity in a measuring device according to the invention.\",\n \"Furthermore for example the cross section of the flexible tube may according to the invention be oval or round, square or rectangular or alternatively have a cavity with a cross section with rounded ends.\",\n \"Also the disclosed capacitive method of measuring the changes of displacements may be substituted by other displacement measuring methods, for example eddy current measurements where the capacitive electrodes are simply substituted by small coils.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476932"}}},{"rowIdx":987,"cells":{"text":{"kind":"list like","value":[["Container positioner","A container positioner assembly for use in positioning a volume of material in a container relative to a dispensing apparatus.","The container positioner assembly includes structures for engagement with the apparatus allowing relative movement of a container attached thereto.","The container positioner assembly may include an assist system which facilitates biased movement of the container relative to the apparatus."],["1.A container positioner assembly for use with beverage preparation equipment comprising, a pair of support guide rails, each configured to be positioned relative to generally opposite sides of a container and each having a pair of tracking slots configured to engage at least one guider roller and to provide a detent position, the guide rails being configured for maintaining an associated container in a generally upright position, at least one guide roller for each slot, a support roller operable to engage an exterior edge of a support guide rail, and at least one handle operably connected to the pair of support guide rails.","2.The container positioner of claim 1 further comprising an assist system having: at least one cable portion attached to a biasing member to form an elongated cable-biasing assembly, with one end of the at least one cable-biasing assembly adapted fro attachment to one of the support guide rails and the other end adapted for attachment to a housing.","3.The container positioner of claim 1 further including, a housing having side walls, the guide rollers and at least one support roller being mounted on the housing sidewalls, and the tracking slots being configured so that pulling on the at least one handle moves the slots with respect to the guide rollers and moves the support guide rails in a generally horizontal forward direction and then downwardly and forwardly until further movement is stopped by at least one roller engaging a detent on one of the slots.","4.The container positioner of claim 1 further including a detent in each tracking slot for removably holding the assembly in the detent position.","5.The container positioner of claim 3 further comprising a shelf connecting the support guide rails and a container on the shelf.","6.The container positioner of claim 5 wherein the rollers are positioned to prevent tipping of the shelf and container during movement of the support guide rails in the generally horizontal forward direction and then downwardly and forwardly.","7.The container positioner assembly of claim 5 further comprising an assist system having: at least one cable portion attached to a biasing to form an elongated cable-biasing assembly, at least one cable-biasing assembly attached at one end to one of the support guide rails and attached at the other end to the housing.","8.A beverage making system comprising, a container for containing beverage making substance, a housing for holding the container, and a container positioner assembly for positioning the container in a beverage making position in the housing for receiving, holding and dispensing the beverage making substance, and for moving the container outwardly and downwardly relative to the housing to a position generally outside of the housing to enable removing or adding material to the container.","9.The beverage making system of claim 8 wherein the means for positioning the container in a beverage making position permits dispensing of beverage making substance from the container to a mixing chamber when the container is in the beverage making position and does not permit dispensing from the container to the mixing chamber when the container is in the position outside of the housing.","10.The beverage making system of claim 8 wherein the means for positioning the container in a beverage making position and for moving the container forwardly and downwardly to a position outside of the housing creates stored energy during movement from the beverage making position to the position outside the housing and uses the stored energy to assist in the return movement from the position outside the housing to the beverage making position.","11.A beverage making system comprising: a container for beverage making substance, the container having a bottom and opposing sidewalls, a housing having sidewalls for enclosing the container on at least two sides, the housing having guide rollers and a support roller carried on the sidewalls, a pair of support guide rails, each connected to generally opposite sidewalls of the container and each having a pair of tracking slots with at least one of the guide rollers in each slot, a least one handle operably connected to the pair of support guide rails, the support roller operably to engage an exterior edge of one of the support guide rails, and a shelf connecting the support guide rails and a container on the shelf.","12.The beverage making system of claim 11 comprising a second support roller operable to engage an exterior edge of the other one of the support guide rails.","13.The beverage making system of claim 11 further comprising an assist system having a pair of cable-biasing assemblies, each cable-biasing assemblies having a cable portion attached to a biasing member, with one end of each cable-biasing assemblies attached to one of the support guide rails and the other end attached to the housing.","14.The beverage making system of claim 11 comprising the tracking slots being shaped so that pulling on the at least one handle moves the container and support guide rails in a generally horizontal forward direction and then downwardly and forwardly until further movement is stopped by at least one roller engaging a detent on one of the slots.","15.The beverage making system of claim 14, wherein the rollers are positioned to prevent tipping of the container during movement in the generally horizontal forward direction and then downwardly and forwardly."],["<SOH> BACKGROUND <EOH>This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.","Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.","In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.","Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.","As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.","Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.","Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.","Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.","These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.","To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.","In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process."],["<SOH> SUMMARY <EOH>The present disclosure relates to apparatus for making and dispensing beverages.","The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.","Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.","The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.","In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.","In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.","The apparatus may also include a plurality of conduits for supplying liquid to the assembly.","The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.","Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.","The housing may be positioned behind the beverage dispensing modules.","In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.","With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.","The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.","A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.","The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.","The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.","In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.","In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.","Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived."],["BACKGROUND This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.","Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.","In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.","Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.","As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.","Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.","Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.","Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.","These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.","To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.","In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process.","SUMMARY The present disclosure relates to apparatus for making and dispensing beverages.","The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.","Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.","The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.","In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.","In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.","The apparatus may also include a plurality of conduits for supplying liquid to the assembly.","The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.","Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.","The housing may be positioned behind the beverage dispensing modules.","In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.","With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.","The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.","A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.","The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.","The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.","In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.","In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.","Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived.","BRIEF DESCRIPTION OF THE DRAWINGS The detailed description particularly refers to the accompanying figures in which: FIG.","1 is a partial fragmentary side elevational view of a modular beverage dispensing apparatus; FIG.","2 is a top plan view of the modular beverage dispensing apparatus of FIG.","1; FIG.","3 is a side elevational view of the modular beverage dispensing apparatus of FIGS.","1 and 2; FIG.","4 is a schematic illustration of the modular beverage dispensing apparatus; FIG.","5 is a schematic illustration of one module of the modular beverage dispensing apparatus; FIG.","6 is a diagrammatic illustration of a dispensing and disposal valve configuration; FIG.","7 is a partial fragmentary top plan view taken along 7-7 in FIG.","6; FIG.","8 is a diagrammatic illustration of a liquid flavor distributing system; FIGS.","9 and 10 are diagrammatic illustrations of the arrangement of liquid flavor distributing systems; FIG.","11 is a cross sectional view, schematic in nature, of a portion of a powdered beverage, soluble beverage dispensing system including connections to dispense liquid flavor to the soluble beverage production stream; FIGS.","12 and 13 are diagrammatic illustrations of the liquid flavor distributing system coupled to the soluble beverage dispensing system of FIG.","11; FIG.","14 is a perspective view of a soluble beverage dispenser having a slide-out assembly; FIG.","15 is an enlarged perspective view of the soluble beverage dispenser of FIG.","14 showing a user removing a container from the soluble beverage dispensing device using the slide-out assembly; FIGS.","16 and 17 are enlarged views of portions of the slide-out assembly of the soluble beverage dispenser of FIG.","14; FIG.","18 is a perspective view of a cable and spring leverage system which assists movement of the slide-out assembly relative to the beverage dispensing system; FIGS.","19 and 20 are schematic illustrations of a communication link between a base station and a beverage dispensing device; and FIGS.","21-28 are schematic illustrations of the circuitry associated with the communication system of FIGS.","19 and 20 AND INCLUDE FIG.","22A, FIGS.","22-22D, FIGS.","23-23D, FIG.","24, FIGS.","24A-24B, FIGS.","25A-25B, FIGS.","26A-26, FIG.","27, FIGS.","27A-27C, FIG.","28 and FIGS.","28A-28E.","DESCRIPTION OF THE DRAWINGS While the present disclosure may be susceptible to embodiment in different forms, there is shown in the drawings, and herein will be described in detail, embodiments with the understanding that the present description is to be considered an exemplification of the principles of the disclosure and is not intended to limit the disclosure to the details of construction and the arrangements of components set forth in the following description or illustrated in the drawings.","A beverage dispensing apparatus 30 illustrated generally in FIGS.","1-4 as comprising a plurality of beverage dispensing modules 32 and a housing 34 that defines a chamber 36 containing water or other liquid suitable for use in brewing.","The housing 34 desirably acts as a central reservoir for a constant supply of heated water to be used by the beverage dispensing modules 32 to brew several different beverages.","The beverage dispensing apparatus 30 may have any suitable construction and configuration.","The beverage apparatus 30 may include any suitable number of beverage dispensing modules 32 oriented in any suitable manner, depending on various circumstances, such as, for example, customer demand, customer usage, available store space, and desired aesthetics.","Similarly, the housing 34 may have any suitable construction and configuration that desirably is consistent with customer demand, customer usage, available store space and desired aesthetics.","A plurality of conduits in the form of water lines 38 provide flow communication between the housing 34 and the beverage dispensing modules 32 to supply water for brewing.","In a preferred embodiment, the housing 34 is positioned behind the beverage dispensing modules 32 and centrally located relative to the beverage dispensing modules 32 to minimize the distance between the housing and each of the beverage dispensing modules, thereby increasing operational efficiency and minimizing space.","Terms including beverage and beverage making as used herein are intended to be broadly defined as including but not limited to the brewing of coffee, tea and any other brewed beverage, including a final beverage or food product as well as producing an intermediate product to be combined with a final beverage or food product.","This broad interpretation is also intended to include, but is not limited to any process of infusing, steeping, reconstituting, diluting, dissolving, saturating or passing a liquid through or otherwise mixing or combining a beverage substance with a liquid such as water without a limitation to the temperature of such liquid unless specified.","This broad interpretation is also intended to include, but is not limited to beverage substances such as ground coffee, tea, liquid beverage concentrate, powdered beverage concentrate, freeze dried coffee or other beverage concentrates, to obtain a desired beverage or other food.","In the illustrated embodiment, for example, the beverage dispensing apparatus 30 comprises ten beverage dispensing modules 32 oriented in a generally U-shaped manner that substantially surrounds the housing 34.As illustrated, four of the beverage dispensing modules 32 face the same direction, four other beverage dispensing modules 32 face an opposite direction, and two of the beverage dispensing modules 32 are positioned in a space-efficient manner consistent with a desired orientation.","A simplified form of the beverage apparatus 30 is shown in FIG.","4 which uses simplified block diagram illustrations to show multiple beverage dispensing modules 32 which are configured to provide a beverage brewing system.","The beverage apparatus 30 shown in FIG.","4 is very similar to a soluble beverage apparatus 30a also shown in FIG.","4.The beverage apparatus 30, as will be described in greater detail herein below, relates to a beverage brewing or producing system whereas the soluble beverage apparatus 30a, as will be described in greater detail herein below, relates to a soluble or powdered beverage dispensing system.","Both beverage apparatuses 30, 30a employ a common heated water reservoir contained in housing 34 to which the individual modules 32, 32a are connected.","As shown in FIGS.","2-4, the housing 34 desirably is centrally located to serve multiple beverage modules 32, 32a.","The benefit of a common housing 34 functioning as a common reservoir to supply water, instead of individual tanks for each module 32, 32a, is that it substantially reduces the cost and greatly increases the efficiency of the system.","Also it reduces the redundancy of multiple heated water reservoirs, reduces the wattage required by the system and increases flexibility in the number of modules 32, 32a which might be included.","The particular configuration with the housing 34 positioned in close proximity to or between several modules 32, 32a is beneficial in that it also minimizes the length of water lines 38 connecting the housing 34 to the modules 32, 32a.","Additionally, a larger body of water in the housing 34 tends to help maintain the temperature and reduce temperature loss over time as well as maintaining a larger volume of brew water at a desired temperature.","While the housing 34 retained within the beverage apparatus 30 is insulated to reduce heat loss and reduce heat load on the ambient environment, heat which may escape from the housing 34 will generally pass to the modules 32, thereby increasing the efficiency of maintaining the temperature of the beverages retained in the modules 32.Insulation desirably is provided to reduce heat loss to the ambient environment.","This will also help reduce the exertion on the ambient air-conditioning system thereby further increasing efficiency of the present beverage apparatus 30, 30a.","With reference to FIGS.","1-4, each module 32, 32a produces a separate brewed beverage.","The centralization of a housing 34 helps reduce the individual cost associated with the module 32, 32a and improves the ease of manufacturing of the modules.","Also, there is increased commonality of parts associated with this type of system.","The modular configuration of the beverage apparatus 30 using the modules 32, 32a increases the potential variability of the physical size and brewer configuration of the system.","This is beneficial to accommodate the needs of a particular location.","In other words, only a desired or required number of modules 32, 32a will be needed at any given time.","If a location has an increase in demand for brewed or soluble beverages additional modules 32, 32a may be added as necessary for the particular change in demand.","Additionally, with regard to repair and maintenance, the beverage apparatus 30, 30a can be designed such that individual modules 32, 32a can be removed for repair and maintenance as necessary with an identical module being inserted in the missing modules.","In a preferred embodiment, (FIG.","5) the housing 34 includes a heating element 52, and one or more system sensors such as a level sensor 86 and a temperature sensor 84.Such components tend to be generally readily available and easily replaced and as such further enhance the reliability of the overall beverage apparatus 30, 30a.","With reference to FIG.","3, the line 38 connecting each housing 34 to a brewing portion 40 of the module 32 is shown.","Water 42 retained in the housing 34 can be readily dispensed.","A water inlet line 44 is provided to introduce unheated water 50 to the housing 34.The inlet line 44 runs from an upper portion 46 of the housing 34 and directs the inward water downwardly towards a bottom portion 48 of the housing 34.Introducing the unheated water 50 towards the bottom of the reservoir introduces the unheated water 50 in close relation to a heated element 52 thereby accelerating the heating of the water.","Additionally, the introduction of unheated water 50 in the bottom portion 48 tends to force heated water in the upper portion 46 upwardly.","As such, the water 42 in the upper portion 46 tends to be the hottest water in the housing 34.Introducing unheated water 50 from the upper portion has the benefit of providing an entrance into the reservoir through a reservoir wall 56 with little or no pressure on the entry therethrough.","This may result in an increase in the reliability of the entry therethrough.","Additionally, a reinforcing or strengthening structure 58 is provided extending between opposed walls 60, 62 of the housing 34.This helps prevent bowing or stress on the walls 60, 62 which might otherwise result from the force of the water on the walls.","Multiple reinforcing structures 58 may be provided throughout the housing 34 as necessary to provide support for the size and dimension of the housing required for a particular system.","FIG.","5, for example, shows a diagrammatic illustration of an embodiment of the apparatus 30 shown in FIGS.","1-4.The beverage dispensing modules 32 can have any suitable construction desirably adapted to produce beverages, store beverages for ready consumption, and facilitate ready dispensing of the beverages.","Each module 32 includes a brewing assembly 30 that may have any suitable construction that can be used to produce beverage such as brewing coffee or other beverages.","The illustrated brewing assembly 30, for example, includes a brewing substance retainer or assembly 64 including a funnel 66.Such a retainer 64 may be well known in the art and may include a filter (not shown) retained in the funnel 66 for holding a charge of beverage substance.","The funnel 66 may be positioned above a reservoir or container portion 68 of the dispensing module 32 which receives brewed beverage from the funnel.","A dispensing assembly 70 in the form of a controllable valve may be associated with the container 68 for dispensing beverage from the module 32.With reference to FIG.","5 a simplified review of the beverage brewing process is described.","A brew actuator switch 72 desirably is associated with each of the modules 32.To initiate a brew cycle for one of the modules 32 a respective brew activation switch 72 is operated.","Operation of the switch 72 sends a signal over line 74 to controller 76.Controller 76 operates an inlet valve 78 over line 80 to introduce water to the housing 34.Also, heating element 52 is coupled to the controller 76 over line 82.The temperature sensor 84 and level sensor 86 are coupled to the controller over lines 88, 90 respectively.","If the controller 76 detects proper level and temperature it will initiate a brew cycle thereby displacing water from the housing 34 through a sprayhead assembly 92.The sprayhead assembly 92 communicates with the substance retainer 64 and dispenses water into the funnel 66 for producing a beverage therein.","The illustrated module 32 includes a level sensor 94 associated with the container 68 which is coupled to the control over line 96.The level sensor can detect a change in level in beverage retained in the container 68 which may be used to activate a brew cycle.","Also, the level detector 94 may sense the level in the beverage container 68 thereby indicating if the container is full or above the desired level to prevent initiation of a brew cycle.","This will prevent over flow of the beverage server 68.The beverage dispensing modules 32 can be constructed to dispense beverage in any suitable manner.","In a preferred embodiment, for example, a dispenser or faucet switch 98 may be associated with the dispensing assembly 70 and coupled with a dispensing handle or control 100.Activation of the control 100 will activate the switch 98 over line 102 connected to the controller 76 thereby indicating that the controller 76 should operate a solenoid valve 104 (FIGS.","6 and 7), via line 106 to dispense beverage therefrom.","As will be described in greater detail below with regard to FIGS.","6 and 7, a disposal or dump valve 108 is also coupled with the container 68 to facilitate controllable disposal, via line 110 coupled to the controller 76.With reference to FIGS.","6 and 7, the beverage container 68 may further include a manifold 114.The illustrated manifold 114 includes a primary passage 116 which communicates with an interior cavity or reservoir 118 of the container 68.Beverage flows from the cavity 118 through the primary passage 116.The solenoid valves 104, 108 communicate with the passage 116.A dispensing passage 120 and a disposal passage 122 communicate with the primary passage 116.Solenoid valve 104 and 108 desirably are normally closed and require specific activation in order to open the valves.","During a dispense cycle the switch or control 100, for example a controllable faucet as shown in FIG.","5, is activated thereby activating solenoid valve 104.When the dispense solenoid valve 104 is opened beverage 124 will flow from the cavity through the solenoid valve 104.Under this condition disposal valve 108 is closed.","When beverage must be disposed of, the disposal solenoid valve 108 is opened while maintaining the dispense solenoid valve 104 in a closed position.","This will allow the disposal of beverage through a separate dispensing line.","Disposal of beverage may occur as a result of a time lapsed period whereby after a specified period of time beverage should be disposed of.","For example, if a beverage only maintains a desired flavor profile or other characteristics for a period of time, for example 2 hours, at the end of two hours after the beverage brewing cycle is initiated, the beverage may be disposed of through the disposal valve 108.Such disposal may also be used at the end of a service cycle such that at the end of the day all beverages may be disposed of at a predetermined time.","The disposal valve may also be used in a manner to flush and cleanse the beverage brewing system.","In this regard, at a predetermined period of time, the controller 76 may operate the disposal valve 108 to dispose of any beverage retained in the container 68.The disposal valve 108 and dispense valve 104 may be closed.","The controller 76 may then dispense a quantity of water into the cavity 118 thereby flushing and sanitizing the reservoir.","After holding the heated water for a predetermined period of time the water may be disposed through the disposal valve 108.Multiple rinsing or flushing cycles may be controlled by the controller 76.In this regard, both the cavity 118, the container 68 and the associated passages 116, 122, 120 and solenoid valve 104, 108 may be cleansed and sanitize periodically and automatically.","It will be appreciated that these automatic cycles will likely greatly increase the cleanliness, taste reliability and sanitation of such beverage dispensing systems as such procedures require considerable amount of time on the part of manual operators.","The use of controllable solenoid valves 104, 108 also prevents dispensing of beverage during a brew cycle.","For example, once a brew cycle is initiated the controller 76 can lock out the solenoid valves 104, 108.This will allow for the brewing cycle to complete and brewed beverage to mix within the container 68 to insure consistent flavor.","The beverage dispensing modules 32 can be constructed in any other suitable manner to dispense brewed beverage in any suitable manner in accordance with various embodiments disclosed.","If desired, for example, the beverage containers 68 of the beverage dispensing modules 32 can instead be in the form of carafes (not shown) or have any other suitable construction.","In a preferred embodiment, during a brewing cycle, heated water 134 is dispensed from the housing 34 through the water dispensing system.","Heated water 134 is used to infuse a beverage brewing substance 136 retained in the brewing assembly 64.An opening 138 is provided in the bottom of the funnel 66 to allow a brewed beverage 140 to flow therefrom.","As shown, for example, in previous figures, a brewed beverage container 68 is provided to collect the beverage 140.The beverage container 68 may be of any suitable form.","With reference to FIG.","8, a liquid flavor dispensing system 130 may be employed with the modular beverage dispensing system disclosed.","The liquid flavor dispensing system 130 can have any suitable configuration and can be used to supply flavoring 132 to all or less than all of the beverage dispensing modules 32.The liquid dispensing system 130 may also be used to supply a different flavoring 132 to each of the different beverage dispensing modules 32.Additionally, the flavorings 132 can be supplied to the beverage dispensing modules by a single liquid flavor dispensing system 130 or several liquid flavor dispensing systems.","Further, the flavoring 132 may in the form of the typical flavorings associated with brewed coffee or instead be in any other suitable form such as, for example, honey, peach, lemon, herbal flavors, or any other suitable flavoring which might be desired by a beverage consumer for any number of different beverages.","In a preferred embodiment, a small quantity of flavoring 132 is introduced into the water which is dispensed to the beverage brewing substance 136.The beverage brewing substance may be in the form of decaff or regular ground coffee as well as tea substances or other substances which might be used in an infusion beverage brewing process.","The objective in using the liquid flavoring material 132 is to introduce a small quantity of concentrated material or substance 132 during the brewing process so that the flavor becomes fully mixed with and saturated in the heated water 134.Introduction of the flavor 132 early in the brewing process will help assure that the flavor fully mixes with the water, is fully flushed through the beverage brewing substance to maximize the value of the flavor and is used to produce an aroma which might entice purchasers of the beverage as part of the beverage purchasing experience.","The flavor could also be dispensed into the beverage brewing substance and subsequently mixed with the water and the resultant brew during the brewing process.","A benefit of adding flavors at the time of brewing and/or mixing soluble beverages, over using pre-flavor beverage substances, is the increased effectiveness of the flavorings.","The increased effectiveness of the flavorings reduces the amount of flavoring required to produce a desired taste profile.","Under these circumstances, the flavorings are mixed with the beverage substance under optimal conditions to maximize the flavoring benefits of the flavorings.","For example, the time prior to mixing the flavoring with the brewing substance is nominal thereby virtually eliminating any degradation of the flavoring which might occur if the flavoring was exposed to air.","A liquid flavor dispensing system 130 is illustrated diagrammatically in FIG.","8.With this embodiment, the liquid flavor dispensing system dispenses a different liquid flavoring 132 in a controlled manner to several of the brewing assemblies of the beverage dispensing modules 32.The flavor can be dispensed to a water or liquid distribution system which controllably dispenses water to a beverage making substance, the beverage making substance or both.","With reference to FIGS.","8-10, the illustrated liquid flavor dispensing system 130 desirably includes a plurality of packages or containers 144a-144h, each of which retains a respective liquid flavor 132 therein.","The containers 144 desirably are in controlled communication with respective beverage dispensing modules 32a-32d in any suitable manner.","For example, a section of tubing or other passage 146 may connect each container 144 to a respective check valve 148.The check valve 148 is part of a respective solenoid or other form of pump device 150 coupled to the controller 76 via a respective line 152.A respective small diameter tube 154 extends from the solenoid pump 150 to a respective second check valve or flavor injection port 156 at a respective terminal end 158 of the tube 154.A respective sensor 160 can be coupled to the controller via a respective line 162 to monitor and detect a low level of the respective flavoring 132 in the respective container 144.As will be described in greater detail below with regard to sensing and monitoring, the status of the flavorings 132 can be reported by the controller 176.The check valves 148 and 156 at each end of each of the tubes 154 provide a sealed line to which the container 144 can be attached.","Also, either alternatively or in addition to the use of the sensor 160, the flavor usage can be inferred based on information collected on a characteristic of the pump such as operation or usage for example, pump actuation.","For example, a sensor may be associated with the pump and run time of the pump 150 could be monitored and a calculation made to infer the amount of flavorings remaining in a corresponding flavorings container 144.The system would provide a signal to the operator, such as “check-status” when the level of flavorings approaches, attains or drops below a predetermined level, for example 20% remaining.","Such advance warning would give the operator the opportunity to change the flavorings container 144 before it runs out.","The advance warning also helps assure that a reasonable margin of error in the inferred calculation is considered and accommodated for.","In use, when a brewing cycle is initiated, the solenoid pump 150 can be activated simultaneous with the initiation of the brewing cycle of one of the beverage dispensing modules or shortly thereafter.","A quantity, for example 9 ml for use in brewing ½ gallon of coffee, of liquid flavoring 132 is pumped from the container 144 through the tubing 154.The check valve or flavor injection port 156 is positioned in close proximity to a sprayhead 170 of the dispensing assembly 92.Dispensing of the flavor 132 into the water will allow the flavor to mix with the heated water for brewing substance during the saturation and extraction process.","Generally, it is desirable to use the flavoring at the start of the cycle which will result in completely integrating the flavor during the brewing cycle.","The process may be repeated with other beverage dispensing modules 32, 32a and, if desired, with other flavors.","The flavor injection ports 156 may, for example, include a check valve of a type as produced by Smart Products.","This is a positive pressure inverted check valve which requires positive pressure through the tube 154 to open the valve.","Positioning of the injection port 156 in close proximity to the sprayhead 170 results in the benefit of clearing and rinsing the surface as a result of the water vapor and heat during a brewing cycle.","Periodic manual cleaning of the sprayhead 170 and injection port 156 helps maintain a clean system.","Desirably, because the injection port 156 is an inverted pressure check valve, physical contact with the tip of the injection port 156 through which the liquid 132 is dispensed does not result in dispensing of liquid 132.Rather, physical contact with the end of the injection port 156 causes the valve to close as opposed to open.","The system also prevents the entry of atmosphere into the container 144 or line 154 thereby preventing contamination or degrading of the flavor or quality of the flavorings.","This system provides a generally closed system which prevents the flavorings contact with the atmosphere and prevents contamination providing a generally aseptic environment.","With reference to FIG.","11, the liquid flavor dispensing system 130 as described with regard to FIGS.","8-10 may be used with a soluble beverage system 176.In FIG.","11, the liquid dispensing system dispenses liquid flavor into a hot water path 174 of a soluble or powdered beverage dispensing system 176.The devices used to dispense the liquid flavor are the same as those describe above with regard to FIG.","8.As such, only the terminal ends 158a, 158b and 158c of multiple lines 154a, 154b, and 154c are shown.","Similarly, the multiple injection ports 156a, 156b, and 156c are shown.","The soluble beverage dispensing system 176 retains a quantity of powdered beverage mix (not shown) in a beverage dispensing container 178.The dispensing of soluble beverage material in powder form is generally known in the art.","The container 178 includes a dispensing auger (not shown) which controllably dispenses powder by operation of a dispensing drive or motor 180 controlled by the controller 76a.","Powdered beverage material is dispensed through the passage 182 into a mixing chamber 184.The hot water line 174 communicates with the mixing chamber 184 to combine with the powder.","The combined powder and hot water flow into an agitating chamber 186 there below.","A whipper motor 188 and agitating blade 190 further blend the powder and water combination for dispensing a beverage 192 through a nozzle 194.The flavor dispensing system 130 (see, FIG.","8) controllably dispenses liquid flavor 132 from respective containers 144 using corresponding pumps via the lines 154a, 154b, and 154c.","Either individual flavors or combinations of flavors may be dispensed into the hot water line.","Such dispensing may be programmed in the controller 76a.","Positioning of the injection ports 156a, 156b, 156c in the heated water path helps to assure that the liquid flavors are rinsed from the injection ports during and at the conclusion of the heated water dispensing cycle during a beverage dispensing cycle.","It should be noted that generally these powdered beverage dispensing systems have a momentary hot water overrun at the end of the cycle during which powdered beverages dispense from the container 178.The slight overrun of heated water helps to assure that the mixing chamber 184, whipping chamber 186, and nozzle 194 are rinsed clean at the end of the cycle.","The injection ports 156a, 156b and 156c have been positioned in the hot water line to provide the rinsing characteristics and benefits of the overrun of hot water.","Furthermore, it is commonly known to have at least one daily rinse cycle in which heated water is dispensed through the assembly 176 to maintain cleanliness and sanitation of the system.","During a brew cycle as shown in FIG.","8, liquid flavor is generally preferably dispensed at the initiation of the brew cycle.","As noted, this helps to ensure complete mixing and optimal extraction of the flavor from the beverage brewing substance 136.A predetermined volume of liquid flavor 132 can be dispensed into the brewing assembly 30 at the initiation of the brew cycle.","In contrast, the soluble system 176 as shown in FIG.","11 may have a variable quantity of beverage 192 dispensed therefrom.","It is anticipated that multiple beverage sizes may be dispensed from the soluble system 176.As a result, the liquid flavoring 132 must be dispensed at a rate consistent with the concentration flavor profile or recipe, and powder also dispensed by the system.","For example, if a consumer wishes to obtain a 12 ounce beverage the proper amount of liquid flavoring must be dispensed for a 12 ounce beverage.","However, the consumer may be allowed to control whether they wish to have a full 12 ounces of beverage or if they decide to short the beverage resulting in, perhaps, only 10 ounces being dispensed.","As such, the pump associated with the powdered beverage system 176 must be calibrated to dispense small quantities of liquid flavoring 132 on a per unit time basis to assure a desirable and consistent mixing of flavor with the powder.","This will help assure that the desired flavor is achieved regardless of the quantity of beverage dispensed.","The solenoid pump 150 used with the liquid flavor dispensing system 130 is a precision controlled metering pump.","It is anticipated that other forms of the metering pump may be used with this system.","However, in the present embodiment, the metering pump is controllably operated to dispense very small quantities of the liquid flavor 132.The frequency of the plunger rate, the stroke of the plunger as well as the duration of plunger operation the solenoid pump 150 can be controlled by the controller 76, 76a.","As a result, very small quantities of highly concentrated liquid flavor can be dispensed into the beverages 140, 192 during the corresponding beverage preparation process.","As such, precise quantities can be added to provide flexibility and tailoring of the resultant beverages.","The system 130 can be calibrated using volumetric guidelines associated with various beverage recipes and flavor materials to match particular flavor preferences, for example for various demographics.","As such, a franchise operation can custom blend and configure the flavor of beverages dispensed by the beverage dispensing system to meet and satisfy regional preferences.","Additionally, if preferences change over time the system can be modified and calibrated to match those preferences.","This adds further flexibility to the beverage system disclosed.","With reference to FIGS.","14-18, a powdered or soluble beverage dispensing system 199 is shown.","While a soluable beverage system 199 has been shown and described as a powder dispensing system, it is envisioned that such a soluable system also includes a liquid concentrate system which dispenses a quality of liquid coffee concentrate for mixing with water or other dilution or reconsituting substance.","Reference to a soluable beverage system is intended to be broadly interpreted.","Such a beverage dispensing system has been generally and diagrammatically shown in FIG.","11.With further reference to FIG.","14, the container 178 is shown positioned inside a housing 200.The passage 182 coupled to the container 178 is shown positioned for dispensing into the mixing chamber 184 and communicating with the whipping chamber 186 and nozzle 194.A portion of the housing 200 in the form of a door 202 conceals the powdered beverage system 176 in the housing 200.Also, the liquid flavor dispensing system 130 as described hereinabove may be used with the powdered beverage or soluble beverage dispensing system 176 illustrated in FIGS.","14-18.One of the potential difficulties involved with a soluble beverage dispensing system is that the container 178 must be removed periodically for refilling with powdered or soluble beverage substance as well as regular maintenance and cleaning.","The container 178 may not be very heavy when it is empty because it is generally formed of a plastic material.","However, once filled, it may contain 5 to 10 pounds of powder material.","With the container 178 being positioned in an elevated location and containing a quantity of material, it may be difficult to lift the container into a desired position in the housing 200.The container 178 is positioned in an upper position in the housing 200 in order to benefit from gravity assistance when dispensing powdered beverage therefrom to the mixing chamber 84.Additionally, it is important to align the dispensing passage 182 with the housing 184 to help assure that when powdered beverage material is dispensed from the container 178 it flows into the proper path for mixing with water.","As such, it would be desirable to provide an apparatus and system for facilitating improved ease of removal and replacement of the powdered beverage container 178 relative to the housing 200.As shown in FIGS.","14-18, a container positioning assembly or container positioner 220 in the form of a slide-out support or shelf is provided.","The container positioning assembly 220 includes a pair of handles 222 which can be grasped (see FIG.","15) by a user to extract the container 178 attached to the assembly 220 for removing it from the housing 200.As shown in FIG.","15, the assembly 220 allows the container 178 to be pulled straight out of the housing 200 thereby allowing disengagement of the passage 182 relative to the mixing chamber 184.Movement of the assembly 220 also replaces the container 178 in a desired position to align the dispensing passage 182 with the mixing chamber 184.As can be seen in FIGS.","15-17, the container 178 has been removed from the housing 200 in order to refill the container 178 but does not have to be removed from the shelf 250 of the assembly 220.Support rails 226 are positioned on each side of container 178 and are engaged with side walls 228 of the housing 200 (FIG.","15).","Each of the guide rails 226 includes a pair of tracking slots 230, 232 which are engaged with corresponding upper 234 and lower 236 guide rollers.","A support roller 240 is positioned along an exterior edge 242 of the guide 226 to further help facilitate smooth and controlled movement of the assembly 220.Support rollers 240 and guide rollers 234, 236 are mounted on the sidewalls of the housing 200.The guides 226 are connected by a cross extending shelf 250 on which the container 178 is positioned.","The combination of the guide rails 226 and rollers help the assembly 220 slide outwardly and inwardly relative to the housing 200.FIG.","16 shows only the portion of the guide rails 226 extending past the edge of container 178.The guide rollers 240, 234, 236 associated with the guides 226 prevent tipping of the shelf 250 and associated container 178.When the assembly 220 is stowed in the housing 200 the shelf 250 rests on a corresponding surface 252 of the housing 200.As can be seen in FIGS.","15-17, the guides 226 include a generally horizontal portion 254 and a vertically oriented portion 256.The shelf 250 and container 178 are associated with the generally horizontal portion 254.The generally vertical portion 256 helps to allow forward movement as well as downward movement of the assembly 220 relative to the housing 200.The guide including the portion 254, 256 help facilitate movement of the container into and out of the housing 200 and forward and downward movement.","In the embodiment as shown, the assembly facilitates positioning of the container 178 approximately 6-8 inches downwardly from its stored position in the housing 200.This lowering of the container 178 is beneficial in that it allows for easy filling of the container.","Also, by fixing the container to the shelf 250 and the assembly 200 it prevents spilling or otherwise tipping of the container 178 during filling or other processes.","This can result in savings in terms of lost beverage material as well as cleanup costs and associated complications.","In the stored position, detents 260, 262 are provided on the guides 230, 232, respectively.","The detents 260, 262 allow the rollers 234, 236 respectively, to be engaged therewith to prevent casual disengagement of the assembly 220 from the housing 200.This helps to lock or retain the assembly 220 in a desired stored or stowed position in the housing 200.As the assembly 220 is moved outwardly from the housing 200, the track guides 234, 236 track along the corresponding channels 230, 232 generally horizontally along the horizontal portion 254.As such, the container 278 attached to the shelf 250 sides forwardly and outwardly from the housing.","At a position where the generally horizontal portion 254 transitions into the partially vertical portion 256 (elbow 270) the guides allow the assembly to slide downwardly and forwardly.","In the extended most position the guides 234, 236 engage corresponding extended detents 272, 274, respectively.","These detent positions 272, 274 help retain the assembly 220 in the outward or extended most position.","The assembly 220 also includes an assist system 280 which includes cables 282 and biasing members to springs 284 forming a cable-biasing assembly help facilitate removal of and replacement of the container 178 relative to the housing 200.With reference to FIG.","18, the assist system 280 includes a spring 284 attached at one 286 to a back portion of the housing and at a second end to a first pulley 288.The cable 282 is attached at a first end 290 to a corresponding portion of the housing and extends through the first pulley 288 and up and around a second pulley 292.The second end 294 of the cable 282 attaches to a corresponding portion of the guides 226 at attachment point 296.Attachment of the cable 282 to the guide 226 is shown in FIG.","16.The assist assembly 288 helps further facilitate ease of removal and replacement of the container 178 relative to the housing 200.For example, when the container 178 is removed from the housing on the assembly 220 the spring 284 stores energy and is stretched as the cables 282 are extended.","The cables 282 are extended as a result of the forward and downward movement of the guides 226.Extension of the springs 284 expends the stored energy and imparts a return force in the springs which helps reduce the force required to return the container 178 to the housing 200.This is particularly useful when an empty container is removed from the housing, filled with powdered material and then returned.","Returning the filled container 178 normally would require additional effort on the part of the operator and as such the return forces in the assist system 280 help make the return easier.","Turning now to FIGS.","19 and 20, the beverage system also includes a communications system or means 300 for communicating 302 between a corresponding beverage brewing, beverage making apparatus or beverage dispensing modules 32, 32a and a receiver or base station 304.Each of the modules 32, 32a includes a controller 76, 76a which is centralized for use with multiple modules or dedicated to an individual module.","The controller collects information about the module or modules.","For purposes of this present discussion, we will refer to the controller as being dedicated to multiple modules and receiving information from each individual module associated therewith.","The many modules associated with a single control also provides the added benefit of the modular assembly as described hereinabove.","It should also be noted that the details of the circuitry used to achieve the communication system are illustrated in the detailed schematics provided in FIGS.","21-28 and all corresponding sub-portions thereof.","For example, FIG.","21 is a general schematic of the overall system.","FIGS.","22-28 provide details about the general system shown in FIG.","21.FIG.","21 has been noted with cross reference to the other figures which provide details about the portions of the circuitry shown generally in FIG.","21.The controllers 76, 76a include the link 302 which connects the controller to the corresponding receiver or base station 304.In the present embodiment as diagrammatically illustrated in FIGS.","19 and 20, the controllers 76, 76a communicate with the receiver 304 by way of an RF link 302.Generally, the controllers are remote from the receiver and use the Link 302 and the associated means for linking to communicate the collected information about the beverage making apparatus.","Antennae 306, 308 are provided on the controllers 76, 76a and the corresponding receivers 304 respectively.","It should be noted that the communication link 302 may be in any one of a variety of forms such as hardwired physical point-to-point link, optical lines, light wave, ultrasonic, infrared or any other form of communication link between one or more devices.","The receiver 304 receives information from the controller or various components of the modules 32, 32a to identify information relating to the modules and operation of the modules.","The receiver 304 may be in the form of a monitoring unit positioned in an appropriate location relative to an operator or attendant who has responsibility for or otherwise maintains the modules 32, 32a.","The receiver 304 includes a means for displaying display 305 and can provide visual auditor or other information about numerous conditions.","The display 304 allows a form of the collected information about the beverage making apparatus to be displayed at the receiver.","The visual display may include, but is not limited to lights, text, symbolic images (i.e., dispenser “full” showing a colored or shaded dispenser and a dispenser “empty” showing an unshaded dispenser), and mechanical devices that tare operated or other visual displays.","An auditory display may include, but is not limited to speech information, alarms, tones or other signals that van be heard.","Tactile displays are also contemplated in the form of a vibrating surface, vibrating device worn by the operator or other means for letting the operator or other means for letting the operator know about he collected information.","For example, the following is a list of the messages or conditions which can be displayed: Receiver fault; Please wait attempting communication; Communication failure, coffee module #; Communication fault, soluble module #; Flavor [type] low, coffee module #; Flavor [type] low, soluble module #; Hopper low, soluble module #; Hopper empty, soluble module #; Coffee empty, coffee module #; Coffee low, coffee module #; Freshness expired, coffee module #; Dumping in ______ minutes, coffee unit #; Server removed, coffee module #; Refill too long; Temperature probe open; Temperature probe short; Heating too long; Over flow safety; Coffee stations all OK; and Soluble stations all OK. For example, in an convenience store setting, multiple modules 32, 32a may be generally remotely positioned in one portion of the store, for example, a rearward location of the store, if desired.","The operator of the beverage system may also be the same person responsible for stocking of the equipment, and register activities.","As such, one person is required to operate and maintain many components of the convenience store facility.","The receiver 304 may be positioned at the cash register so that while the attendant is operating the cash register he may also be notified of matters that require his attention at the modules 32, 32a.","For example, if a module 32a indicates that a container 68 is empty the controller 76 will communicate with the receiver 304 to indicate this condition.","The receiver may also be configured with audible alarms and visual displays to provide additional information.","For example, a display may identify which module and the specific condition associated with the module which has provided a signal to the receiver 304.The operator can then make a decision how to use that information relative to his responsibilities at the cash register.","If an emergency requires immediate attention he can secure the cash register and address the emergency while attending to the module 32.Alternatively, if it is a slow period in the store and a module is indicating that a container 68 is empty he can put off attending to this condition until he completes one or more transactions in queue.","The signal communication between the controller 76 and the receiver 304 may be a single path signal or in the form of a multiple path verification signal.","For example, as shown in FIGS.","19 and 20 a first signal 310 is transmitted from the controller 76 to the receiver 304.This initial communication may report the status of a condition at the module 32.The receiver 304 can then return a copy of the information 312 to the controller 76.If the controller confirms the information received (312) it can then send a return confirmation 314 that the information is correct.","This is a useful communication link in the present situation due to the presence of electrical noise and other interference within other beverage system applications.","For example, each of the components associated with the modules 32, 32a may provide some degree of electrical or other noise.","Additionally, other systems within the beverage system setting may be producing RF or other signals which could create interference.","As such, the present embodiment providing the verification communication links 310-314 helps facilitate and assure accurate and timely communication.","In the event of excessive interference such as from patrons of the store using telephone or RF communication devices, the controller 76 can continue to periodically send a message 310 to the receiver 304 until the verification signal 312 is received.","While preferred embodiments are disclosed, illustrated and described, it is envisioned that those skilled in the art may devise various modifications and equivalents without departing from the spirit and scope of the disclosure as recited in the following claims."]],"string":"[\n [\n \"Container positioner\",\n \"A container positioner assembly for use in positioning a volume of material in a container relative to a dispensing apparatus.\",\n \"The container positioner assembly includes structures for engagement with the apparatus allowing relative movement of a container attached thereto.\",\n \"The container positioner assembly may include an assist system which facilitates biased movement of the container relative to the apparatus.\"\n ],\n [\n \"1.A container positioner assembly for use with beverage preparation equipment comprising, a pair of support guide rails, each configured to be positioned relative to generally opposite sides of a container and each having a pair of tracking slots configured to engage at least one guider roller and to provide a detent position, the guide rails being configured for maintaining an associated container in a generally upright position, at least one guide roller for each slot, a support roller operable to engage an exterior edge of a support guide rail, and at least one handle operably connected to the pair of support guide rails.\",\n \"2.The container positioner of claim 1 further comprising an assist system having: at least one cable portion attached to a biasing member to form an elongated cable-biasing assembly, with one end of the at least one cable-biasing assembly adapted fro attachment to one of the support guide rails and the other end adapted for attachment to a housing.\",\n \"3.The container positioner of claim 1 further including, a housing having side walls, the guide rollers and at least one support roller being mounted on the housing sidewalls, and the tracking slots being configured so that pulling on the at least one handle moves the slots with respect to the guide rollers and moves the support guide rails in a generally horizontal forward direction and then downwardly and forwardly until further movement is stopped by at least one roller engaging a detent on one of the slots.\",\n \"4.The container positioner of claim 1 further including a detent in each tracking slot for removably holding the assembly in the detent position.\",\n \"5.The container positioner of claim 3 further comprising a shelf connecting the support guide rails and a container on the shelf.\",\n \"6.The container positioner of claim 5 wherein the rollers are positioned to prevent tipping of the shelf and container during movement of the support guide rails in the generally horizontal forward direction and then downwardly and forwardly.\",\n \"7.The container positioner assembly of claim 5 further comprising an assist system having: at least one cable portion attached to a biasing to form an elongated cable-biasing assembly, at least one cable-biasing assembly attached at one end to one of the support guide rails and attached at the other end to the housing.\",\n \"8.A beverage making system comprising, a container for containing beverage making substance, a housing for holding the container, and a container positioner assembly for positioning the container in a beverage making position in the housing for receiving, holding and dispensing the beverage making substance, and for moving the container outwardly and downwardly relative to the housing to a position generally outside of the housing to enable removing or adding material to the container.\",\n \"9.The beverage making system of claim 8 wherein the means for positioning the container in a beverage making position permits dispensing of beverage making substance from the container to a mixing chamber when the container is in the beverage making position and does not permit dispensing from the container to the mixing chamber when the container is in the position outside of the housing.\",\n \"10.The beverage making system of claim 8 wherein the means for positioning the container in a beverage making position and for moving the container forwardly and downwardly to a position outside of the housing creates stored energy during movement from the beverage making position to the position outside the housing and uses the stored energy to assist in the return movement from the position outside the housing to the beverage making position.\",\n \"11.A beverage making system comprising: a container for beverage making substance, the container having a bottom and opposing sidewalls, a housing having sidewalls for enclosing the container on at least two sides, the housing having guide rollers and a support roller carried on the sidewalls, a pair of support guide rails, each connected to generally opposite sidewalls of the container and each having a pair of tracking slots with at least one of the guide rollers in each slot, a least one handle operably connected to the pair of support guide rails, the support roller operably to engage an exterior edge of one of the support guide rails, and a shelf connecting the support guide rails and a container on the shelf.\",\n \"12.The beverage making system of claim 11 comprising a second support roller operable to engage an exterior edge of the other one of the support guide rails.\",\n \"13.The beverage making system of claim 11 further comprising an assist system having a pair of cable-biasing assemblies, each cable-biasing assemblies having a cable portion attached to a biasing member, with one end of each cable-biasing assemblies attached to one of the support guide rails and the other end attached to the housing.\",\n \"14.The beverage making system of claim 11 comprising the tracking slots being shaped so that pulling on the at least one handle moves the container and support guide rails in a generally horizontal forward direction and then downwardly and forwardly until further movement is stopped by at least one roller engaging a detent on one of the slots.\",\n \"15.The beverage making system of claim 14, wherein the rollers are positioned to prevent tipping of the container during movement in the generally horizontal forward direction and then downwardly and forwardly.\"\n ],\n [\n \"<SOH> BACKGROUND <EOH>This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.\",\n \"Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.\",\n \"In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.\",\n \"Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.\",\n \"As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.\",\n \"Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.\",\n \"Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.\",\n \"Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.\",\n \"These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.\",\n \"To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.\",\n \"In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process.\"\n ],\n [\n \"<SOH> SUMMARY <EOH>The present disclosure relates to apparatus for making and dispensing beverages.\",\n \"The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.\",\n \"Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.\",\n \"The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.\",\n \"In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.\",\n \"In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.\",\n \"The apparatus may also include a plurality of conduits for supplying liquid to the assembly.\",\n \"The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.\",\n \"Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.\",\n \"The housing may be positioned behind the beverage dispensing modules.\",\n \"In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.\",\n \"With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.\",\n \"The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.\",\n \"A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.\",\n \"The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.\",\n \"The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.\",\n \"In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.\",\n \"In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.\",\n \"Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived.\"\n ],\n [\n \"BACKGROUND This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.\",\n \"Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.\",\n \"In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.\",\n \"Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.\",\n \"As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.\",\n \"Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.\",\n \"Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.\",\n \"Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.\",\n \"These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.\",\n \"To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.\",\n \"In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process.\",\n \"SUMMARY The present disclosure relates to apparatus for making and dispensing beverages.\",\n \"The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.\",\n \"Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.\",\n \"The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.\",\n \"In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.\",\n \"In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.\",\n \"The apparatus may also include a plurality of conduits for supplying liquid to the assembly.\",\n \"The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.\",\n \"Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.\",\n \"The housing may be positioned behind the beverage dispensing modules.\",\n \"In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.\",\n \"With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.\",\n \"The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.\",\n \"A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.\",\n \"The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.\",\n \"The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.\",\n \"In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.\",\n \"In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.\",\n \"Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS The detailed description particularly refers to the accompanying figures in which: FIG.\",\n \"1 is a partial fragmentary side elevational view of a modular beverage dispensing apparatus; FIG.\",\n \"2 is a top plan view of the modular beverage dispensing apparatus of FIG.\",\n \"1; FIG.\",\n \"3 is a side elevational view of the modular beverage dispensing apparatus of FIGS.\",\n \"1 and 2; FIG.\",\n \"4 is a schematic illustration of the modular beverage dispensing apparatus; FIG.\",\n \"5 is a schematic illustration of one module of the modular beverage dispensing apparatus; FIG.\",\n \"6 is a diagrammatic illustration of a dispensing and disposal valve configuration; FIG.\",\n \"7 is a partial fragmentary top plan view taken along 7-7 in FIG.\",\n \"6; FIG.\",\n \"8 is a diagrammatic illustration of a liquid flavor distributing system; FIGS.\",\n \"9 and 10 are diagrammatic illustrations of the arrangement of liquid flavor distributing systems; FIG.\",\n \"11 is a cross sectional view, schematic in nature, of a portion of a powdered beverage, soluble beverage dispensing system including connections to dispense liquid flavor to the soluble beverage production stream; FIGS.\",\n \"12 and 13 are diagrammatic illustrations of the liquid flavor distributing system coupled to the soluble beverage dispensing system of FIG.\",\n \"11; FIG.\",\n \"14 is a perspective view of a soluble beverage dispenser having a slide-out assembly; FIG.\",\n \"15 is an enlarged perspective view of the soluble beverage dispenser of FIG.\",\n \"14 showing a user removing a container from the soluble beverage dispensing device using the slide-out assembly; FIGS.\",\n \"16 and 17 are enlarged views of portions of the slide-out assembly of the soluble beverage dispenser of FIG.\",\n \"14; FIG.\",\n \"18 is a perspective view of a cable and spring leverage system which assists movement of the slide-out assembly relative to the beverage dispensing system; FIGS.\",\n \"19 and 20 are schematic illustrations of a communication link between a base station and a beverage dispensing device; and FIGS.\",\n \"21-28 are schematic illustrations of the circuitry associated with the communication system of FIGS.\",\n \"19 and 20 AND INCLUDE FIG.\",\n \"22A, FIGS.\",\n \"22-22D, FIGS.\",\n \"23-23D, FIG.\",\n \"24, FIGS.\",\n \"24A-24B, FIGS.\",\n \"25A-25B, FIGS.\",\n \"26A-26, FIG.\",\n \"27, FIGS.\",\n \"27A-27C, FIG.\",\n \"28 and FIGS.\",\n \"28A-28E.\",\n \"DESCRIPTION OF THE DRAWINGS While the present disclosure may be susceptible to embodiment in different forms, there is shown in the drawings, and herein will be described in detail, embodiments with the understanding that the present description is to be considered an exemplification of the principles of the disclosure and is not intended to limit the disclosure to the details of construction and the arrangements of components set forth in the following description or illustrated in the drawings.\",\n \"A beverage dispensing apparatus 30 illustrated generally in FIGS.\",\n \"1-4 as comprising a plurality of beverage dispensing modules 32 and a housing 34 that defines a chamber 36 containing water or other liquid suitable for use in brewing.\",\n \"The housing 34 desirably acts as a central reservoir for a constant supply of heated water to be used by the beverage dispensing modules 32 to brew several different beverages.\",\n \"The beverage dispensing apparatus 30 may have any suitable construction and configuration.\",\n \"The beverage apparatus 30 may include any suitable number of beverage dispensing modules 32 oriented in any suitable manner, depending on various circumstances, such as, for example, customer demand, customer usage, available store space, and desired aesthetics.\",\n \"Similarly, the housing 34 may have any suitable construction and configuration that desirably is consistent with customer demand, customer usage, available store space and desired aesthetics.\",\n \"A plurality of conduits in the form of water lines 38 provide flow communication between the housing 34 and the beverage dispensing modules 32 to supply water for brewing.\",\n \"In a preferred embodiment, the housing 34 is positioned behind the beverage dispensing modules 32 and centrally located relative to the beverage dispensing modules 32 to minimize the distance between the housing and each of the beverage dispensing modules, thereby increasing operational efficiency and minimizing space.\",\n \"Terms including beverage and beverage making as used herein are intended to be broadly defined as including but not limited to the brewing of coffee, tea and any other brewed beverage, including a final beverage or food product as well as producing an intermediate product to be combined with a final beverage or food product.\",\n \"This broad interpretation is also intended to include, but is not limited to any process of infusing, steeping, reconstituting, diluting, dissolving, saturating or passing a liquid through or otherwise mixing or combining a beverage substance with a liquid such as water without a limitation to the temperature of such liquid unless specified.\",\n \"This broad interpretation is also intended to include, but is not limited to beverage substances such as ground coffee, tea, liquid beverage concentrate, powdered beverage concentrate, freeze dried coffee or other beverage concentrates, to obtain a desired beverage or other food.\",\n \"In the illustrated embodiment, for example, the beverage dispensing apparatus 30 comprises ten beverage dispensing modules 32 oriented in a generally U-shaped manner that substantially surrounds the housing 34.As illustrated, four of the beverage dispensing modules 32 face the same direction, four other beverage dispensing modules 32 face an opposite direction, and two of the beverage dispensing modules 32 are positioned in a space-efficient manner consistent with a desired orientation.\",\n \"A simplified form of the beverage apparatus 30 is shown in FIG.\",\n \"4 which uses simplified block diagram illustrations to show multiple beverage dispensing modules 32 which are configured to provide a beverage brewing system.\",\n \"The beverage apparatus 30 shown in FIG.\",\n \"4 is very similar to a soluble beverage apparatus 30a also shown in FIG.\",\n \"4.The beverage apparatus 30, as will be described in greater detail herein below, relates to a beverage brewing or producing system whereas the soluble beverage apparatus 30a, as will be described in greater detail herein below, relates to a soluble or powdered beverage dispensing system.\",\n \"Both beverage apparatuses 30, 30a employ a common heated water reservoir contained in housing 34 to which the individual modules 32, 32a are connected.\",\n \"As shown in FIGS.\",\n \"2-4, the housing 34 desirably is centrally located to serve multiple beverage modules 32, 32a.\",\n \"The benefit of a common housing 34 functioning as a common reservoir to supply water, instead of individual tanks for each module 32, 32a, is that it substantially reduces the cost and greatly increases the efficiency of the system.\",\n \"Also it reduces the redundancy of multiple heated water reservoirs, reduces the wattage required by the system and increases flexibility in the number of modules 32, 32a which might be included.\",\n \"The particular configuration with the housing 34 positioned in close proximity to or between several modules 32, 32a is beneficial in that it also minimizes the length of water lines 38 connecting the housing 34 to the modules 32, 32a.\",\n \"Additionally, a larger body of water in the housing 34 tends to help maintain the temperature and reduce temperature loss over time as well as maintaining a larger volume of brew water at a desired temperature.\",\n \"While the housing 34 retained within the beverage apparatus 30 is insulated to reduce heat loss and reduce heat load on the ambient environment, heat which may escape from the housing 34 will generally pass to the modules 32, thereby increasing the efficiency of maintaining the temperature of the beverages retained in the modules 32.Insulation desirably is provided to reduce heat loss to the ambient environment.\",\n \"This will also help reduce the exertion on the ambient air-conditioning system thereby further increasing efficiency of the present beverage apparatus 30, 30a.\",\n \"With reference to FIGS.\",\n \"1-4, each module 32, 32a produces a separate brewed beverage.\",\n \"The centralization of a housing 34 helps reduce the individual cost associated with the module 32, 32a and improves the ease of manufacturing of the modules.\",\n \"Also, there is increased commonality of parts associated with this type of system.\",\n \"The modular configuration of the beverage apparatus 30 using the modules 32, 32a increases the potential variability of the physical size and brewer configuration of the system.\",\n \"This is beneficial to accommodate the needs of a particular location.\",\n \"In other words, only a desired or required number of modules 32, 32a will be needed at any given time.\",\n \"If a location has an increase in demand for brewed or soluble beverages additional modules 32, 32a may be added as necessary for the particular change in demand.\",\n \"Additionally, with regard to repair and maintenance, the beverage apparatus 30, 30a can be designed such that individual modules 32, 32a can be removed for repair and maintenance as necessary with an identical module being inserted in the missing modules.\",\n \"In a preferred embodiment, (FIG.\",\n \"5) the housing 34 includes a heating element 52, and one or more system sensors such as a level sensor 86 and a temperature sensor 84.Such components tend to be generally readily available and easily replaced and as such further enhance the reliability of the overall beverage apparatus 30, 30a.\",\n \"With reference to FIG.\",\n \"3, the line 38 connecting each housing 34 to a brewing portion 40 of the module 32 is shown.\",\n \"Water 42 retained in the housing 34 can be readily dispensed.\",\n \"A water inlet line 44 is provided to introduce unheated water 50 to the housing 34.The inlet line 44 runs from an upper portion 46 of the housing 34 and directs the inward water downwardly towards a bottom portion 48 of the housing 34.Introducing the unheated water 50 towards the bottom of the reservoir introduces the unheated water 50 in close relation to a heated element 52 thereby accelerating the heating of the water.\",\n \"Additionally, the introduction of unheated water 50 in the bottom portion 48 tends to force heated water in the upper portion 46 upwardly.\",\n \"As such, the water 42 in the upper portion 46 tends to be the hottest water in the housing 34.Introducing unheated water 50 from the upper portion has the benefit of providing an entrance into the reservoir through a reservoir wall 56 with little or no pressure on the entry therethrough.\",\n \"This may result in an increase in the reliability of the entry therethrough.\",\n \"Additionally, a reinforcing or strengthening structure 58 is provided extending between opposed walls 60, 62 of the housing 34.This helps prevent bowing or stress on the walls 60, 62 which might otherwise result from the force of the water on the walls.\",\n \"Multiple reinforcing structures 58 may be provided throughout the housing 34 as necessary to provide support for the size and dimension of the housing required for a particular system.\",\n \"FIG.\",\n \"5, for example, shows a diagrammatic illustration of an embodiment of the apparatus 30 shown in FIGS.\",\n \"1-4.The beverage dispensing modules 32 can have any suitable construction desirably adapted to produce beverages, store beverages for ready consumption, and facilitate ready dispensing of the beverages.\",\n \"Each module 32 includes a brewing assembly 30 that may have any suitable construction that can be used to produce beverage such as brewing coffee or other beverages.\",\n \"The illustrated brewing assembly 30, for example, includes a brewing substance retainer or assembly 64 including a funnel 66.Such a retainer 64 may be well known in the art and may include a filter (not shown) retained in the funnel 66 for holding a charge of beverage substance.\",\n \"The funnel 66 may be positioned above a reservoir or container portion 68 of the dispensing module 32 which receives brewed beverage from the funnel.\",\n \"A dispensing assembly 70 in the form of a controllable valve may be associated with the container 68 for dispensing beverage from the module 32.With reference to FIG.\",\n \"5 a simplified review of the beverage brewing process is described.\",\n \"A brew actuator switch 72 desirably is associated with each of the modules 32.To initiate a brew cycle for one of the modules 32 a respective brew activation switch 72 is operated.\",\n \"Operation of the switch 72 sends a signal over line 74 to controller 76.Controller 76 operates an inlet valve 78 over line 80 to introduce water to the housing 34.Also, heating element 52 is coupled to the controller 76 over line 82.The temperature sensor 84 and level sensor 86 are coupled to the controller over lines 88, 90 respectively.\",\n \"If the controller 76 detects proper level and temperature it will initiate a brew cycle thereby displacing water from the housing 34 through a sprayhead assembly 92.The sprayhead assembly 92 communicates with the substance retainer 64 and dispenses water into the funnel 66 for producing a beverage therein.\",\n \"The illustrated module 32 includes a level sensor 94 associated with the container 68 which is coupled to the control over line 96.The level sensor can detect a change in level in beverage retained in the container 68 which may be used to activate a brew cycle.\",\n \"Also, the level detector 94 may sense the level in the beverage container 68 thereby indicating if the container is full or above the desired level to prevent initiation of a brew cycle.\",\n \"This will prevent over flow of the beverage server 68.The beverage dispensing modules 32 can be constructed to dispense beverage in any suitable manner.\",\n \"In a preferred embodiment, for example, a dispenser or faucet switch 98 may be associated with the dispensing assembly 70 and coupled with a dispensing handle or control 100.Activation of the control 100 will activate the switch 98 over line 102 connected to the controller 76 thereby indicating that the controller 76 should operate a solenoid valve 104 (FIGS.\",\n \"6 and 7), via line 106 to dispense beverage therefrom.\",\n \"As will be described in greater detail below with regard to FIGS.\",\n \"6 and 7, a disposal or dump valve 108 is also coupled with the container 68 to facilitate controllable disposal, via line 110 coupled to the controller 76.With reference to FIGS.\",\n \"6 and 7, the beverage container 68 may further include a manifold 114.The illustrated manifold 114 includes a primary passage 116 which communicates with an interior cavity or reservoir 118 of the container 68.Beverage flows from the cavity 118 through the primary passage 116.The solenoid valves 104, 108 communicate with the passage 116.A dispensing passage 120 and a disposal passage 122 communicate with the primary passage 116.Solenoid valve 104 and 108 desirably are normally closed and require specific activation in order to open the valves.\",\n \"During a dispense cycle the switch or control 100, for example a controllable faucet as shown in FIG.\",\n \"5, is activated thereby activating solenoid valve 104.When the dispense solenoid valve 104 is opened beverage 124 will flow from the cavity through the solenoid valve 104.Under this condition disposal valve 108 is closed.\",\n \"When beverage must be disposed of, the disposal solenoid valve 108 is opened while maintaining the dispense solenoid valve 104 in a closed position.\",\n \"This will allow the disposal of beverage through a separate dispensing line.\",\n \"Disposal of beverage may occur as a result of a time lapsed period whereby after a specified period of time beverage should be disposed of.\",\n \"For example, if a beverage only maintains a desired flavor profile or other characteristics for a period of time, for example 2 hours, at the end of two hours after the beverage brewing cycle is initiated, the beverage may be disposed of through the disposal valve 108.Such disposal may also be used at the end of a service cycle such that at the end of the day all beverages may be disposed of at a predetermined time.\",\n \"The disposal valve may also be used in a manner to flush and cleanse the beverage brewing system.\",\n \"In this regard, at a predetermined period of time, the controller 76 may operate the disposal valve 108 to dispose of any beverage retained in the container 68.The disposal valve 108 and dispense valve 104 may be closed.\",\n \"The controller 76 may then dispense a quantity of water into the cavity 118 thereby flushing and sanitizing the reservoir.\",\n \"After holding the heated water for a predetermined period of time the water may be disposed through the disposal valve 108.Multiple rinsing or flushing cycles may be controlled by the controller 76.In this regard, both the cavity 118, the container 68 and the associated passages 116, 122, 120 and solenoid valve 104, 108 may be cleansed and sanitize periodically and automatically.\",\n \"It will be appreciated that these automatic cycles will likely greatly increase the cleanliness, taste reliability and sanitation of such beverage dispensing systems as such procedures require considerable amount of time on the part of manual operators.\",\n \"The use of controllable solenoid valves 104, 108 also prevents dispensing of beverage during a brew cycle.\",\n \"For example, once a brew cycle is initiated the controller 76 can lock out the solenoid valves 104, 108.This will allow for the brewing cycle to complete and brewed beverage to mix within the container 68 to insure consistent flavor.\",\n \"The beverage dispensing modules 32 can be constructed in any other suitable manner to dispense brewed beverage in any suitable manner in accordance with various embodiments disclosed.\",\n \"If desired, for example, the beverage containers 68 of the beverage dispensing modules 32 can instead be in the form of carafes (not shown) or have any other suitable construction.\",\n \"In a preferred embodiment, during a brewing cycle, heated water 134 is dispensed from the housing 34 through the water dispensing system.\",\n \"Heated water 134 is used to infuse a beverage brewing substance 136 retained in the brewing assembly 64.An opening 138 is provided in the bottom of the funnel 66 to allow a brewed beverage 140 to flow therefrom.\",\n \"As shown, for example, in previous figures, a brewed beverage container 68 is provided to collect the beverage 140.The beverage container 68 may be of any suitable form.\",\n \"With reference to FIG.\",\n \"8, a liquid flavor dispensing system 130 may be employed with the modular beverage dispensing system disclosed.\",\n \"The liquid flavor dispensing system 130 can have any suitable configuration and can be used to supply flavoring 132 to all or less than all of the beverage dispensing modules 32.The liquid dispensing system 130 may also be used to supply a different flavoring 132 to each of the different beverage dispensing modules 32.Additionally, the flavorings 132 can be supplied to the beverage dispensing modules by a single liquid flavor dispensing system 130 or several liquid flavor dispensing systems.\",\n \"Further, the flavoring 132 may in the form of the typical flavorings associated with brewed coffee or instead be in any other suitable form such as, for example, honey, peach, lemon, herbal flavors, or any other suitable flavoring which might be desired by a beverage consumer for any number of different beverages.\",\n \"In a preferred embodiment, a small quantity of flavoring 132 is introduced into the water which is dispensed to the beverage brewing substance 136.The beverage brewing substance may be in the form of decaff or regular ground coffee as well as tea substances or other substances which might be used in an infusion beverage brewing process.\",\n \"The objective in using the liquid flavoring material 132 is to introduce a small quantity of concentrated material or substance 132 during the brewing process so that the flavor becomes fully mixed with and saturated in the heated water 134.Introduction of the flavor 132 early in the brewing process will help assure that the flavor fully mixes with the water, is fully flushed through the beverage brewing substance to maximize the value of the flavor and is used to produce an aroma which might entice purchasers of the beverage as part of the beverage purchasing experience.\",\n \"The flavor could also be dispensed into the beverage brewing substance and subsequently mixed with the water and the resultant brew during the brewing process.\",\n \"A benefit of adding flavors at the time of brewing and/or mixing soluble beverages, over using pre-flavor beverage substances, is the increased effectiveness of the flavorings.\",\n \"The increased effectiveness of the flavorings reduces the amount of flavoring required to produce a desired taste profile.\",\n \"Under these circumstances, the flavorings are mixed with the beverage substance under optimal conditions to maximize the flavoring benefits of the flavorings.\",\n \"For example, the time prior to mixing the flavoring with the brewing substance is nominal thereby virtually eliminating any degradation of the flavoring which might occur if the flavoring was exposed to air.\",\n \"A liquid flavor dispensing system 130 is illustrated diagrammatically in FIG.\",\n \"8.With this embodiment, the liquid flavor dispensing system dispenses a different liquid flavoring 132 in a controlled manner to several of the brewing assemblies of the beverage dispensing modules 32.The flavor can be dispensed to a water or liquid distribution system which controllably dispenses water to a beverage making substance, the beverage making substance or both.\",\n \"With reference to FIGS.\",\n \"8-10, the illustrated liquid flavor dispensing system 130 desirably includes a plurality of packages or containers 144a-144h, each of which retains a respective liquid flavor 132 therein.\",\n \"The containers 144 desirably are in controlled communication with respective beverage dispensing modules 32a-32d in any suitable manner.\",\n \"For example, a section of tubing or other passage 146 may connect each container 144 to a respective check valve 148.The check valve 148 is part of a respective solenoid or other form of pump device 150 coupled to the controller 76 via a respective line 152.A respective small diameter tube 154 extends from the solenoid pump 150 to a respective second check valve or flavor injection port 156 at a respective terminal end 158 of the tube 154.A respective sensor 160 can be coupled to the controller via a respective line 162 to monitor and detect a low level of the respective flavoring 132 in the respective container 144.As will be described in greater detail below with regard to sensing and monitoring, the status of the flavorings 132 can be reported by the controller 176.The check valves 148 and 156 at each end of each of the tubes 154 provide a sealed line to which the container 144 can be attached.\",\n \"Also, either alternatively or in addition to the use of the sensor 160, the flavor usage can be inferred based on information collected on a characteristic of the pump such as operation or usage for example, pump actuation.\",\n \"For example, a sensor may be associated with the pump and run time of the pump 150 could be monitored and a calculation made to infer the amount of flavorings remaining in a corresponding flavorings container 144.The system would provide a signal to the operator, such as “check-status” when the level of flavorings approaches, attains or drops below a predetermined level, for example 20% remaining.\",\n \"Such advance warning would give the operator the opportunity to change the flavorings container 144 before it runs out.\",\n \"The advance warning also helps assure that a reasonable margin of error in the inferred calculation is considered and accommodated for.\",\n \"In use, when a brewing cycle is initiated, the solenoid pump 150 can be activated simultaneous with the initiation of the brewing cycle of one of the beverage dispensing modules or shortly thereafter.\",\n \"A quantity, for example 9 ml for use in brewing ½ gallon of coffee, of liquid flavoring 132 is pumped from the container 144 through the tubing 154.The check valve or flavor injection port 156 is positioned in close proximity to a sprayhead 170 of the dispensing assembly 92.Dispensing of the flavor 132 into the water will allow the flavor to mix with the heated water for brewing substance during the saturation and extraction process.\",\n \"Generally, it is desirable to use the flavoring at the start of the cycle which will result in completely integrating the flavor during the brewing cycle.\",\n \"The process may be repeated with other beverage dispensing modules 32, 32a and, if desired, with other flavors.\",\n \"The flavor injection ports 156 may, for example, include a check valve of a type as produced by Smart Products.\",\n \"This is a positive pressure inverted check valve which requires positive pressure through the tube 154 to open the valve.\",\n \"Positioning of the injection port 156 in close proximity to the sprayhead 170 results in the benefit of clearing and rinsing the surface as a result of the water vapor and heat during a brewing cycle.\",\n \"Periodic manual cleaning of the sprayhead 170 and injection port 156 helps maintain a clean system.\",\n \"Desirably, because the injection port 156 is an inverted pressure check valve, physical contact with the tip of the injection port 156 through which the liquid 132 is dispensed does not result in dispensing of liquid 132.Rather, physical contact with the end of the injection port 156 causes the valve to close as opposed to open.\",\n \"The system also prevents the entry of atmosphere into the container 144 or line 154 thereby preventing contamination or degrading of the flavor or quality of the flavorings.\",\n \"This system provides a generally closed system which prevents the flavorings contact with the atmosphere and prevents contamination providing a generally aseptic environment.\",\n \"With reference to FIG.\",\n \"11, the liquid flavor dispensing system 130 as described with regard to FIGS.\",\n \"8-10 may be used with a soluble beverage system 176.In FIG.\",\n \"11, the liquid dispensing system dispenses liquid flavor into a hot water path 174 of a soluble or powdered beverage dispensing system 176.The devices used to dispense the liquid flavor are the same as those describe above with regard to FIG.\",\n \"8.As such, only the terminal ends 158a, 158b and 158c of multiple lines 154a, 154b, and 154c are shown.\",\n \"Similarly, the multiple injection ports 156a, 156b, and 156c are shown.\",\n \"The soluble beverage dispensing system 176 retains a quantity of powdered beverage mix (not shown) in a beverage dispensing container 178.The dispensing of soluble beverage material in powder form is generally known in the art.\",\n \"The container 178 includes a dispensing auger (not shown) which controllably dispenses powder by operation of a dispensing drive or motor 180 controlled by the controller 76a.\",\n \"Powdered beverage material is dispensed through the passage 182 into a mixing chamber 184.The hot water line 174 communicates with the mixing chamber 184 to combine with the powder.\",\n \"The combined powder and hot water flow into an agitating chamber 186 there below.\",\n \"A whipper motor 188 and agitating blade 190 further blend the powder and water combination for dispensing a beverage 192 through a nozzle 194.The flavor dispensing system 130 (see, FIG.\",\n \"8) controllably dispenses liquid flavor 132 from respective containers 144 using corresponding pumps via the lines 154a, 154b, and 154c.\",\n \"Either individual flavors or combinations of flavors may be dispensed into the hot water line.\",\n \"Such dispensing may be programmed in the controller 76a.\",\n \"Positioning of the injection ports 156a, 156b, 156c in the heated water path helps to assure that the liquid flavors are rinsed from the injection ports during and at the conclusion of the heated water dispensing cycle during a beverage dispensing cycle.\",\n \"It should be noted that generally these powdered beverage dispensing systems have a momentary hot water overrun at the end of the cycle during which powdered beverages dispense from the container 178.The slight overrun of heated water helps to assure that the mixing chamber 184, whipping chamber 186, and nozzle 194 are rinsed clean at the end of the cycle.\",\n \"The injection ports 156a, 156b and 156c have been positioned in the hot water line to provide the rinsing characteristics and benefits of the overrun of hot water.\",\n \"Furthermore, it is commonly known to have at least one daily rinse cycle in which heated water is dispensed through the assembly 176 to maintain cleanliness and sanitation of the system.\",\n \"During a brew cycle as shown in FIG.\",\n \"8, liquid flavor is generally preferably dispensed at the initiation of the brew cycle.\",\n \"As noted, this helps to ensure complete mixing and optimal extraction of the flavor from the beverage brewing substance 136.A predetermined volume of liquid flavor 132 can be dispensed into the brewing assembly 30 at the initiation of the brew cycle.\",\n \"In contrast, the soluble system 176 as shown in FIG.\",\n \"11 may have a variable quantity of beverage 192 dispensed therefrom.\",\n \"It is anticipated that multiple beverage sizes may be dispensed from the soluble system 176.As a result, the liquid flavoring 132 must be dispensed at a rate consistent with the concentration flavor profile or recipe, and powder also dispensed by the system.\",\n \"For example, if a consumer wishes to obtain a 12 ounce beverage the proper amount of liquid flavoring must be dispensed for a 12 ounce beverage.\",\n \"However, the consumer may be allowed to control whether they wish to have a full 12 ounces of beverage or if they decide to short the beverage resulting in, perhaps, only 10 ounces being dispensed.\",\n \"As such, the pump associated with the powdered beverage system 176 must be calibrated to dispense small quantities of liquid flavoring 132 on a per unit time basis to assure a desirable and consistent mixing of flavor with the powder.\",\n \"This will help assure that the desired flavor is achieved regardless of the quantity of beverage dispensed.\",\n \"The solenoid pump 150 used with the liquid flavor dispensing system 130 is a precision controlled metering pump.\",\n \"It is anticipated that other forms of the metering pump may be used with this system.\",\n \"However, in the present embodiment, the metering pump is controllably operated to dispense very small quantities of the liquid flavor 132.The frequency of the plunger rate, the stroke of the plunger as well as the duration of plunger operation the solenoid pump 150 can be controlled by the controller 76, 76a.\",\n \"As a result, very small quantities of highly concentrated liquid flavor can be dispensed into the beverages 140, 192 during the corresponding beverage preparation process.\",\n \"As such, precise quantities can be added to provide flexibility and tailoring of the resultant beverages.\",\n \"The system 130 can be calibrated using volumetric guidelines associated with various beverage recipes and flavor materials to match particular flavor preferences, for example for various demographics.\",\n \"As such, a franchise operation can custom blend and configure the flavor of beverages dispensed by the beverage dispensing system to meet and satisfy regional preferences.\",\n \"Additionally, if preferences change over time the system can be modified and calibrated to match those preferences.\",\n \"This adds further flexibility to the beverage system disclosed.\",\n \"With reference to FIGS.\",\n \"14-18, a powdered or soluble beverage dispensing system 199 is shown.\",\n \"While a soluable beverage system 199 has been shown and described as a powder dispensing system, it is envisioned that such a soluable system also includes a liquid concentrate system which dispenses a quality of liquid coffee concentrate for mixing with water or other dilution or reconsituting substance.\",\n \"Reference to a soluable beverage system is intended to be broadly interpreted.\",\n \"Such a beverage dispensing system has been generally and diagrammatically shown in FIG.\",\n \"11.With further reference to FIG.\",\n \"14, the container 178 is shown positioned inside a housing 200.The passage 182 coupled to the container 178 is shown positioned for dispensing into the mixing chamber 184 and communicating with the whipping chamber 186 and nozzle 194.A portion of the housing 200 in the form of a door 202 conceals the powdered beverage system 176 in the housing 200.Also, the liquid flavor dispensing system 130 as described hereinabove may be used with the powdered beverage or soluble beverage dispensing system 176 illustrated in FIGS.\",\n \"14-18.One of the potential difficulties involved with a soluble beverage dispensing system is that the container 178 must be removed periodically for refilling with powdered or soluble beverage substance as well as regular maintenance and cleaning.\",\n \"The container 178 may not be very heavy when it is empty because it is generally formed of a plastic material.\",\n \"However, once filled, it may contain 5 to 10 pounds of powder material.\",\n \"With the container 178 being positioned in an elevated location and containing a quantity of material, it may be difficult to lift the container into a desired position in the housing 200.The container 178 is positioned in an upper position in the housing 200 in order to benefit from gravity assistance when dispensing powdered beverage therefrom to the mixing chamber 84.Additionally, it is important to align the dispensing passage 182 with the housing 184 to help assure that when powdered beverage material is dispensed from the container 178 it flows into the proper path for mixing with water.\",\n \"As such, it would be desirable to provide an apparatus and system for facilitating improved ease of removal and replacement of the powdered beverage container 178 relative to the housing 200.As shown in FIGS.\",\n \"14-18, a container positioning assembly or container positioner 220 in the form of a slide-out support or shelf is provided.\",\n \"The container positioning assembly 220 includes a pair of handles 222 which can be grasped (see FIG.\",\n \"15) by a user to extract the container 178 attached to the assembly 220 for removing it from the housing 200.As shown in FIG.\",\n \"15, the assembly 220 allows the container 178 to be pulled straight out of the housing 200 thereby allowing disengagement of the passage 182 relative to the mixing chamber 184.Movement of the assembly 220 also replaces the container 178 in a desired position to align the dispensing passage 182 with the mixing chamber 184.As can be seen in FIGS.\",\n \"15-17, the container 178 has been removed from the housing 200 in order to refill the container 178 but does not have to be removed from the shelf 250 of the assembly 220.Support rails 226 are positioned on each side of container 178 and are engaged with side walls 228 of the housing 200 (FIG.\",\n \"15).\",\n \"Each of the guide rails 226 includes a pair of tracking slots 230, 232 which are engaged with corresponding upper 234 and lower 236 guide rollers.\",\n \"A support roller 240 is positioned along an exterior edge 242 of the guide 226 to further help facilitate smooth and controlled movement of the assembly 220.Support rollers 240 and guide rollers 234, 236 are mounted on the sidewalls of the housing 200.The guides 226 are connected by a cross extending shelf 250 on which the container 178 is positioned.\",\n \"The combination of the guide rails 226 and rollers help the assembly 220 slide outwardly and inwardly relative to the housing 200.FIG.\",\n \"16 shows only the portion of the guide rails 226 extending past the edge of container 178.The guide rollers 240, 234, 236 associated with the guides 226 prevent tipping of the shelf 250 and associated container 178.When the assembly 220 is stowed in the housing 200 the shelf 250 rests on a corresponding surface 252 of the housing 200.As can be seen in FIGS.\",\n \"15-17, the guides 226 include a generally horizontal portion 254 and a vertically oriented portion 256.The shelf 250 and container 178 are associated with the generally horizontal portion 254.The generally vertical portion 256 helps to allow forward movement as well as downward movement of the assembly 220 relative to the housing 200.The guide including the portion 254, 256 help facilitate movement of the container into and out of the housing 200 and forward and downward movement.\",\n \"In the embodiment as shown, the assembly facilitates positioning of the container 178 approximately 6-8 inches downwardly from its stored position in the housing 200.This lowering of the container 178 is beneficial in that it allows for easy filling of the container.\",\n \"Also, by fixing the container to the shelf 250 and the assembly 200 it prevents spilling or otherwise tipping of the container 178 during filling or other processes.\",\n \"This can result in savings in terms of lost beverage material as well as cleanup costs and associated complications.\",\n \"In the stored position, detents 260, 262 are provided on the guides 230, 232, respectively.\",\n \"The detents 260, 262 allow the rollers 234, 236 respectively, to be engaged therewith to prevent casual disengagement of the assembly 220 from the housing 200.This helps to lock or retain the assembly 220 in a desired stored or stowed position in the housing 200.As the assembly 220 is moved outwardly from the housing 200, the track guides 234, 236 track along the corresponding channels 230, 232 generally horizontally along the horizontal portion 254.As such, the container 278 attached to the shelf 250 sides forwardly and outwardly from the housing.\",\n \"At a position where the generally horizontal portion 254 transitions into the partially vertical portion 256 (elbow 270) the guides allow the assembly to slide downwardly and forwardly.\",\n \"In the extended most position the guides 234, 236 engage corresponding extended detents 272, 274, respectively.\",\n \"These detent positions 272, 274 help retain the assembly 220 in the outward or extended most position.\",\n \"The assembly 220 also includes an assist system 280 which includes cables 282 and biasing members to springs 284 forming a cable-biasing assembly help facilitate removal of and replacement of the container 178 relative to the housing 200.With reference to FIG.\",\n \"18, the assist system 280 includes a spring 284 attached at one 286 to a back portion of the housing and at a second end to a first pulley 288.The cable 282 is attached at a first end 290 to a corresponding portion of the housing and extends through the first pulley 288 and up and around a second pulley 292.The second end 294 of the cable 282 attaches to a corresponding portion of the guides 226 at attachment point 296.Attachment of the cable 282 to the guide 226 is shown in FIG.\",\n \"16.The assist assembly 288 helps further facilitate ease of removal and replacement of the container 178 relative to the housing 200.For example, when the container 178 is removed from the housing on the assembly 220 the spring 284 stores energy and is stretched as the cables 282 are extended.\",\n \"The cables 282 are extended as a result of the forward and downward movement of the guides 226.Extension of the springs 284 expends the stored energy and imparts a return force in the springs which helps reduce the force required to return the container 178 to the housing 200.This is particularly useful when an empty container is removed from the housing, filled with powdered material and then returned.\",\n \"Returning the filled container 178 normally would require additional effort on the part of the operator and as such the return forces in the assist system 280 help make the return easier.\",\n \"Turning now to FIGS.\",\n \"19 and 20, the beverage system also includes a communications system or means 300 for communicating 302 between a corresponding beverage brewing, beverage making apparatus or beverage dispensing modules 32, 32a and a receiver or base station 304.Each of the modules 32, 32a includes a controller 76, 76a which is centralized for use with multiple modules or dedicated to an individual module.\",\n \"The controller collects information about the module or modules.\",\n \"For purposes of this present discussion, we will refer to the controller as being dedicated to multiple modules and receiving information from each individual module associated therewith.\",\n \"The many modules associated with a single control also provides the added benefit of the modular assembly as described hereinabove.\",\n \"It should also be noted that the details of the circuitry used to achieve the communication system are illustrated in the detailed schematics provided in FIGS.\",\n \"21-28 and all corresponding sub-portions thereof.\",\n \"For example, FIG.\",\n \"21 is a general schematic of the overall system.\",\n \"FIGS.\",\n \"22-28 provide details about the general system shown in FIG.\",\n \"21.FIG.\",\n \"21 has been noted with cross reference to the other figures which provide details about the portions of the circuitry shown generally in FIG.\",\n \"21.The controllers 76, 76a include the link 302 which connects the controller to the corresponding receiver or base station 304.In the present embodiment as diagrammatically illustrated in FIGS.\",\n \"19 and 20, the controllers 76, 76a communicate with the receiver 304 by way of an RF link 302.Generally, the controllers are remote from the receiver and use the Link 302 and the associated means for linking to communicate the collected information about the beverage making apparatus.\",\n \"Antennae 306, 308 are provided on the controllers 76, 76a and the corresponding receivers 304 respectively.\",\n \"It should be noted that the communication link 302 may be in any one of a variety of forms such as hardwired physical point-to-point link, optical lines, light wave, ultrasonic, infrared or any other form of communication link between one or more devices.\",\n \"The receiver 304 receives information from the controller or various components of the modules 32, 32a to identify information relating to the modules and operation of the modules.\",\n \"The receiver 304 may be in the form of a monitoring unit positioned in an appropriate location relative to an operator or attendant who has responsibility for or otherwise maintains the modules 32, 32a.\",\n \"The receiver 304 includes a means for displaying display 305 and can provide visual auditor or other information about numerous conditions.\",\n \"The display 304 allows a form of the collected information about the beverage making apparatus to be displayed at the receiver.\",\n \"The visual display may include, but is not limited to lights, text, symbolic images (i.e., dispenser “full” showing a colored or shaded dispenser and a dispenser “empty” showing an unshaded dispenser), and mechanical devices that tare operated or other visual displays.\",\n \"An auditory display may include, but is not limited to speech information, alarms, tones or other signals that van be heard.\",\n \"Tactile displays are also contemplated in the form of a vibrating surface, vibrating device worn by the operator or other means for letting the operator or other means for letting the operator know about he collected information.\",\n \"For example, the following is a list of the messages or conditions which can be displayed: Receiver fault; Please wait attempting communication; Communication failure, coffee module #; Communication fault, soluble module #; Flavor [type] low, coffee module #; Flavor [type] low, soluble module #; Hopper low, soluble module #; Hopper empty, soluble module #; Coffee empty, coffee module #; Coffee low, coffee module #; Freshness expired, coffee module #; Dumping in ______ minutes, coffee unit #; Server removed, coffee module #; Refill too long; Temperature probe open; Temperature probe short; Heating too long; Over flow safety; Coffee stations all OK; and Soluble stations all OK. For example, in an convenience store setting, multiple modules 32, 32a may be generally remotely positioned in one portion of the store, for example, a rearward location of the store, if desired.\",\n \"The operator of the beverage system may also be the same person responsible for stocking of the equipment, and register activities.\",\n \"As such, one person is required to operate and maintain many components of the convenience store facility.\",\n \"The receiver 304 may be positioned at the cash register so that while the attendant is operating the cash register he may also be notified of matters that require his attention at the modules 32, 32a.\",\n \"For example, if a module 32a indicates that a container 68 is empty the controller 76 will communicate with the receiver 304 to indicate this condition.\",\n \"The receiver may also be configured with audible alarms and visual displays to provide additional information.\",\n \"For example, a display may identify which module and the specific condition associated with the module which has provided a signal to the receiver 304.The operator can then make a decision how to use that information relative to his responsibilities at the cash register.\",\n \"If an emergency requires immediate attention he can secure the cash register and address the emergency while attending to the module 32.Alternatively, if it is a slow period in the store and a module is indicating that a container 68 is empty he can put off attending to this condition until he completes one or more transactions in queue.\",\n \"The signal communication between the controller 76 and the receiver 304 may be a single path signal or in the form of a multiple path verification signal.\",\n \"For example, as shown in FIGS.\",\n \"19 and 20 a first signal 310 is transmitted from the controller 76 to the receiver 304.This initial communication may report the status of a condition at the module 32.The receiver 304 can then return a copy of the information 312 to the controller 76.If the controller confirms the information received (312) it can then send a return confirmation 314 that the information is correct.\",\n \"This is a useful communication link in the present situation due to the presence of electrical noise and other interference within other beverage system applications.\",\n \"For example, each of the components associated with the modules 32, 32a may provide some degree of electrical or other noise.\",\n \"Additionally, other systems within the beverage system setting may be producing RF or other signals which could create interference.\",\n \"As such, the present embodiment providing the verification communication links 310-314 helps facilitate and assure accurate and timely communication.\",\n \"In the event of excessive interference such as from patrons of the store using telephone or RF communication devices, the controller 76 can continue to periodically send a message 310 to the receiver 304 until the verification signal 312 is received.\",\n \"While preferred embodiments are disclosed, illustrated and described, it is envisioned that those skilled in the art may devise various modifications and equivalents without departing from the spirit and scope of the disclosure as recited in the following claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476949"}}},{"rowIdx":988,"cells":{"text":{"kind":"list like","value":[["Multiple sequencible and ligatible structures for genomic analysis","High throughput methods and kits for single nucleotide polymorphism (SNP) genotyping are provided.","The methods involve utilizing nested PCR amplification reactions which produce sequencible and ligatible structures.","An outer PCR primer set amplifies the SNP, and an inner PCR primer set amplifies a portion of the DNA amplified by the outer primer set, but does not amplify the SNP itself.","The inner and outer primers may reaction include non-target common domain sequences, and the inner primer common domain sequences may comprise digestion restriction endonuclease recognition sites.","The design of the inner primer set allows precise tailoring of the sequencible and ligatible structures with respect to length and base composition."],["1.A method of producing hybrid DNA with a single strand overhang that includes a target site, comprising the steps of: obtaining a first primer which hybridizes to a 5′ strand of a strand of deoxyribonucleic acid (DNA), a second primer which hybridizes to a 3′ strand of said strand of DNA, and a third primer which hydridizes to said 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using said first primer, said second primer, and said third primer, an outer amplicon which includes said target site and an inner amplicon which excludes said target site; and forming at least one of a ligatable structure which includes a 3′-5′ sequence which excludes said target site hybridized to a 5′-3′ sequence which includes said target site, and a sequencible structure which includes a 5′-3′ sequence which excludes said target site hybridized to a 3′-5′ sequence which includes said target site.","2.The method of claim 1 wherein said forming step forms both said ligatible structure and said sequencible structure.","3.The method of claim 1 wherein said target site includes at least one single nucleotide polymorphism.","4.A method of genotyping DNA of an individual by analyzing at least one target site in said DNA, comprising the steps of: obtaining a first primer which hybridizes to a 5′ strand of a strand of said DNA, a second primer which hybridizes to a 3′ strand of said strand of DNA, and a third primer which hydridizes to said 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using said first primer, said second primer, and said third primer, an outer amplicon which includes said target site and an inner amplicon which excludes said target site; and forming at least one of a sequencible structure which includes a 5′-3′ sequence which excludes said target site hybridized to a 3′-5′ sequence which includes said target site; and a ligatable structure which includes a 3′-5′ sequence which excludes said target site hybridized to a 5′-3′ sequence which includes said target site; and analyzing at least one of a sequencing product formed by sequencing said sequencible structure and a ligation product formed by ligating said ligatible structure with a labeled oligonucleotide with a DNA sequencer to determine the genotype of said individual.","5.The method of claim 4 wherein said forming step forms both said ligatible structure and said sequencible structure.","6.The method of claim 4 wherein said at least one target site includes a single nucleotide polymorphism.","7.The method of claim 4 wherein said sequencible structure is sequenced by a technique selected from the group consisting of dideoxy sequencing, pyrosequencing, and single base extension.","8.The method of claim 4 wherein a plurality of target sites are simultaneously analyzed by the step of producing an ordered series of sequencing products of varying, non-overlapping length each specific for a particular target site, and wherein said step of analyzing is carried out by electrophoresing said ordered series of sequencing products in a single channel of said DNA sequencer.","9.The method of claim 8 wherein said step of producing an ordered series of sequencing products is carried out by single base extension (SBE).","10.The method of claim 8 wherein said step of producing an ordered series of sequencing products is carried out by a dideoxy sequencing reaction utilizing a low ratio of dNTPs to ddNTPs.","11.The method of claim 4 wherein said labeled oligonucleotide is fluorescently labeled.","12.The method of claim 4 wherein a plurality of ligatable structures are formed in said forming step, each being specific for one of a plurality of target sites, and said ligation products are of varying, non-overlapping lengths with said labeled oligonucleotides being degenerate, said step of analyzing is carried out by electrophoresing a plurality of ligation products in a single channel of said DNA sequencer.","13.A method for analyzing at least one target site in a DNA molecule, comprising the steps of amplifying by nested PCR said target site, wherein said nested PCR is carried out using inner and outer PCR primer pairs, wherein said outer PCR primer pair forms a first PCR product which contains said target site, and wherein said inner PCR primer pair forms a second PCR product which contains a portion of said first PCR product but does not contain said target site, denaturing said first and said second PCR products to form ssDNA sequences, reannealing said ss DNA sequences to form a sequencible hybrid DNA molecule and a ligatible hybrid DNA molecule, performing sequencing reactions with said sequencible hybrid DNA molecule and ligation reactions with said ligatible hybrid DNA molecule, and determining the characteristics of said target site by analyzing results obtained in said performing step.","14.The method of claim 13 wherein said at least one target site is an SNP polymorphism site.","15.The method of claim 13 wherein said inner and outer primer pairs comprise a sequence tag.","16.The method of claim 15 wherein said sequence tag comprises a restriction enzyme recognition site.","17.The method of claim 13 wherein said step of amplifying is carried out using a low concentration of primers, and further comprising a second step of amplifying, wherein said second step of amplifying uses secondary primers for amplification of said sequence tags.","18.The method of claim 13 wherein one target site is analyzed.","19.The method of claim 13 wherein a plurality of target sites are analyzed.","20.The method of claim 13 wherein said step of amplifying is carried out in a single multiplex PCR reaction.","21.The method of claim 13 wherein said step of amplifying is carried out in multiple independent PCR reactions.","22.The method of claim 13 wherein said results obtained in said performing step are analyzed by a DNA sequencer.","23.Inner and outer PCR primer pairs for the amplification of a target site in a DNA molecule, wherein said outer PCR primer pair forms a first PCR product which contains said target site, and wherein said inner PCR primer pair forms a second PCR product which contains a portion of said first PCR product but does not contain said target site.","24.The primer pairs of claim 23, wherein said target site is an SNP polymorphism site.","25.The primer pairs of claim 23, wherein said primer pairs comprise a sequence tag.","26.A kit for amplification of at least one target site in a DNA molecule, comprising inner and outer PCR primer pairs for the amplification of said target site, wherein said outer primer pair amplifies a portion of said DNA molecule including said target site, and wherein said inner primer pair amplifies part of said portion of said DNA molecule amplified by said outer primer pair and excludes said target site.","27.The kit of claim 26 wherein said inner and outer PCR primer pairs comprise sequence tags.","28.The kit of claim 27, further comprising secondary primers for the amplification of said sequence tags.","29.A dideoxy DNA sequencing kit for producing short chain termination fragments, comprising dNTPs and ddNTPs present in a dNTP:ddNTP ratio ranging from 1:0 to 1:10."],["<SOH> BACKGROUND OF THE INVENTION <EOH>1.Field of the Invention The invention generally relates to single nucleotide polymorphism (SNP) genotyping.","In particular, the invention provides a method for SNP genotyping based on a nested PCR design that creates structures directly suitable for both DNA sequencing and ligation reactions.","2.Background of the Invention With the completion of the rough draft of the human genome, over 1.5 million non-redundant SNPs have been identified and mapped by the publicly funded project and the SNP consortium (Sachidanandam et al, 2001).","The availability of such a large number of SNPs has prompted great interest in SNP related applications and research, such as large scale linkage, genomic association and pharmarcogenetic studies.","SNPs entered the central arena of genetics due to their abundance (Collins et al, 1997; Sachidanandam et al, 2001), stability (Sachidanandam et al, 2001) and the relative ease (Kwok 2000; Shi 2001; Syvanen 2002) with which they are genotyped.","The targets of SNP genotyping are normally small pieces of DNA, ranging from 150-300 base pairs.","Based on the SNP consortium report, the average SNP density in the human genome appears to be greater than 1 SNP/KB.","This high density provides a tool to analyze genomic structure at very high resolution, and to establish a direct correlation between genetic coding variation and biological function.","The relative stability of SNPs (compared to mini- and micro-satellite markers) in evolution makes it simpler to carry out this task.","Although SNP genotyping is generally easier than microsatellite genotyping, and despite the rapid and significant advances in SNP genotyping technology in the last few years, major issues remain unsolved.","Whereas only a few years ago, the main concern was the need to have a large number of SNPs available, currently the most urgent issues are increasing throughput and decreasing cost.","For example, for large scale applications as projected by Kruglyak (1999) and Long (1999), a reasonable cost would be <$0.10/genotype, or else large-scale projects will become prohibitively expensive.","To fulfill the goals of understanding the genetics of complex traits and common diseases, cost-effective and higher throughput methodology in SNP genotyping is essential.","Typically, genotyping protocols have three components: target amplification, allelic discrimination and product detection and identification.","They are normally executed sequentially, but can be processed in a single step reaction, depending on the means which are used for allele discrimination and signal detection.","Such single step procedures are well-suited to automation, but are not necessarily effective in terms of cost and throughput.","PCR is the dominant procedure utilized for target amplification.","As is widely known, a PCR can readily amplify targets present in lower copy number by a factor of 108 or more.","With respect to allele discrimination, all SNP genotyping technologies currently available are based on mechanisms involving one or more of the following: DNA polymerase, hybridization and DNA ligase.","DNA polymerase methods include single base extension (SBE) (Armstrong, 2000; Barta 2001; Bray 2001; Cai 2001; Chen 1997; Chem 1999; Chen 1998; Chen 1997; Chen 1997; Fan 2000; Lindblad-Toh 2000; Nikiforov 1994; Pastinen 1996; Pastinen 2000; Ross, 1998; Sauer 2000; Syvanen 1999; Ye 2001), de novo sequencing including pyrosequencing (Nordstrom 2000; Ronaghi 2001), allele-specific PCR and extension (Germer 1999; Myakishev 2001; Pastinen 2000), and structure-specific cleavage (Fors 2000).","Of these different forms, SBE is the most widely used and has been adapted for many different detection platforms.","Hybridization is also widely used in several different forms, including dynamic hybridization (Prince 2001), and is the primary method currently used in all microarray detection formats.","DNA ligase methods are based on the ability of DNA ligase to join the ends of two oligonucleotides annealed next to each other on a template (Tong 2000; Tong1999).","Two oligonucleotides can be designed to anneal to both sides of a SNP site, and by detecting the formation of ligation product, the genotype of a target can be inferred (Chen 1998; Delahunty 1996; Gerry 1999; Iannone 2000).","Once a target DNA sequence has been amplified and the allelic variants discriminated, the next step in a generic genotyping protocol is to detect and identify the allele specific products.","Detection mechanisms vary greatly, from simple fluorescence intensity (Armstrong 2000; Cai 2000; Chen 1998; Chen 1997; Delahunty 1996; Dubertret 2001; Fan 2000; Fergusoon 2000; Fors 2000; Germer 1999; Germer 2000; Gerry 1999; Iannone 2000; Lindblad-Toh 2000; Lindroos 2001; Marras 1999; Medintz 2001; Myakishev 2001; Nikiforov 1994; Pastinen 1997; Pastinen 2000; Prince 2001; Syvanen 1999; Ye 2001) to very precise mass (Bray 2001; Ross 1998; Sauer 2000) or electric charge (Gilles 1999; Woolley 2000) measurement.","The detection mechanisms roughly fall into two categories: homogeneous and solid phase mediated detection.","All homogeneous detection platforms depend on measuring fluorescence intensities and their change during and/or after reactions.","One common feature among homogeneous approaches is that they do not require any separation/purification prior to signal acquisition, making them amenable to automation.","Solid support techniques include flow cytometry genotyping (Armstrong 2000; Cai 2000; Ye 2001), zip-code microarrays (Fan 2000; Gerry 1999), and mass spectrometry genotyping (Bray 2001; Ross 1998; Sauer 2000).","Using a solid support in the detection step may potentially increase throughput and reduce cost, but unfortunately also entails the risk of complicating protocols and compromising data quality.","For example, when reaction mixtures are applied to a solid support, unintended binding of fluorescence dye to the solid surface can occur, necessitating extensive washing to minimize spurious signals.","With respect to cost, prices for genotyping technologies currently available on the market vary considerably, from $0.50-2.00/genotype.","For example, the template-directed dye-terminator incorporation assay with fluorescence detection (FP-TDI) (Chen 1999) costs roughly $0.50/genotype for reagents by list price.","For other methods, due to their requirement for special reagents and/or clean-up procedures, the cost is higher, e.g.","MALDI-TOF mass spectrometry is about $0.75-1.00, whereas pyrosequencing approaches $2.00/genotype.","No currently available technology approaches the low cost which is necessary in order to prevent large-scale projects from becoming prohibitively expensive, i.e.","<$0.10/genotype.","There are currently many genotyping applications in use or under development that require a relatively large number of both SNPs and samples (e.g., both in the thousands or more), demanding both cost effective and high throughput technologies.","Examples include gene mapping studies by linkage or linkage disequilibrium (Kruglyak 1999; Long 1999) for complex traits and pharmacogenetics (Riley 2000).","Clearly, these applications are critical to improving our understanding of the genetics of complex traits, common diseases and drug response, and to help attain the goal of individualized medicine.","However, none of the current approaches are suitable for such applications in terms of cost effectiveness coupled with high throughput potential, and SNP genotyping demands both."],["<SOH> SUMMARY OF THE INVENTION <EOH>The present invention provides a methodology for high-throughput SNP genotyping that is highly cost effective.","This novel approach makes use of the high throughput capacity of DNA sequencers for SNP genotyping and is based on a nested PCR design that creates a series of ordered “structures” in parallel and exploits them for SNP genotyping assays.","The invention thus provides novel genotyping technologies that significantly increase throughput, reduce cost and are accessible for most researchers, leading to significant improvements in genotyping technology.","While typical SNP genotyping approaches currently available score <10 SNPs simultaneously, the methods of the present invention increase the number to about 50-100 with obvious advantages for throughput and cost.","The present invention provides a method of producing hybrid DNA with a single strand overhang that includes a target sequence.","The steps of the method include: obtaining a first primer which hybridizes to a 5′ strand of a strand of deoxyribonucleic acid (DNA), a second primer which hybridizes to a 3′ strand of the strand of DNA, and a third primer which hydridizes to said 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using the first primer, the second primer, and the third primer, an outer amplicon which includes a target sequence and an inner amplicon which excludes the target sequence; forming at least one of the following: a ligatable structure which includes a 3′-5′ sequence which excludes the target sequence hybridized to a 5′-3′ sequence which includes the target sequence, and a sequencible structure which includes a 5′-3′ sequence which excludes the target sequence hybridized to a 3′-5′ sequence which includes the target sequence; sequencing the sequencible structure(s) and ligating the ligatible structure(s) with a labeled oligonucleotide; and analyzing the sequencing products and ligation products with a DNA sequencer to determine the genotype of said individual.","The method may form both the ligatible structure and the sequencible structure.","The target sequence(s) may include a single nucleotide polymorphism.","The invention also provides a method of genotyping the deoxyribonucleic acid (DNA) of an individual by analyzing at least on target sequence in the DNA.","The steps of the method include: obtaining a first primer which hybridizes to a 5′ strand of a strand of said DNA, a second primer which hybridizes to a 3′ strand of the strand of DNA, and a third primer which hydridizes to the 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using the first primer, the second primer, and the third primer, an outer amplicon which includes said target sequence and an inner amplicon which excludes the target sequence; and forming at least one of the following: a ligatable structure which includes a 3′-5′ sequence which excludes the target sequence hybridized to a 5′-3′ sequence which includes the target sequence, and a sequencible structure which includes a 5′-3′ sequence which excludes the target sequence hybridized to a 3′-5′ sequence which includes the target sequence; sequencing the sequencible structure and ligating the ligatible structure with a labeled oligonucleotide; and analyzing the sequencing and ligation products so produced with a DNA sequencer to determine the genotype of the individual.","The forming step of the method may form both the ligatible structure and the sequencible structure.","The target sequence(s) may include a single nucleotide polymorphism.","The sequencible structure may be sequenced by a technique such as dideoxy sequencing, pyrosequencing, and single base extension.","In a preferred embodiment, of the method, i) a plurality of target sequences is analyzed; ii) the step of forming forms an ordered series of sequencible structures; iii) the step of sequencing produces an ordered series of sequencing products of varying, non-overlapping length; and iv) the step of analyzing is carried out by electrophoresing the ordered series of sequencing products in a single channel of said DNA sequencer.","In some embodiments, the step of sequencing is carried out by single base extension (SBE).","In other embodiments, the step of sequencing is carried out by a dideoxy sequencing reaction utilizing a ratio of dNTPs to ddNTPs that is lower than that which is typically used in order to produce a short sequence reading.","In a preferred embodiment of the present invention, the labeled oligonucleotide is fluorescently labeled.","In one embodiment of the method, i) a plurality of target sequences is analyzed; ii) the labeled oligonucleotides are degenerate; iii) the ligation products are of varying, non-overlapping lengths; and iv) the step of analyzing is carried out by electrophoresing a plurality of ligation products in a single channel of said DNA sequencer.","The present invention also provides a method for analyzing at least one target site in a DNA molecule.","The method includes the steps of: 1) amplifying the target site(s) by nested PCR; (The nested PCR is carried out using inner and outer PCR primer pairs, wherein the outer PCR primer pair forms a first PCR product which contains a target site, and the inner PCR primer pair forms a second PCR product which contains a portion of the first PCR product but does not contain the target site); 2) denaturing the first and said second PCR products to form ssDNA sequences, 3) reannealing the ss DNA sequences to form a sequencible hybrid DNA molecule and a ligatible hybrid DNA molecule, 4) performing sequencing reactions with said sequencible hybrid DNA molecule and ligation reactions with said ligatible hybrid DNA molecule, and 5) determining the characteristics of the target sequence by analyzing results obtained in the performing step.","In a preferred embodiment of the method, the target site(s) may be SNP polymorphism sites.","Further, the inner and outer primer pairs may comprise a sequence tag, which in turn may comprises a restriction enzyme recognition site.","In some embodiments of the method, the step of amplifying may be carried out using a low concentration of primers, and may further comprising a second step of amplification using secondary primers for amplification of the sequence tags.","According to the method, one or several target sites may be analyzed.","The step of amplifying may be carried out in a single multiplex PCR reaction, or in multiple independent PCR reactions, and the results may be analyzed by a DNA sequencer.","The invention also provides inner and outer PCR primer pairs for the amplification of a target sequence in a DNA molecule, wherein the outer PCR primer pair forms a first PCR product which contains the target sequence, and the inner PCR primer pair forms a second PCR product which contains a portion of the first PCR product but does not contain said target sequence.","The target sequence may be an SNP polymorphism site.","The primer pairs may comprise a sequence tag.","The present invention also provides a kit for amplification of at least one target sequence in a DNA molecule.","The kit includes inner and outer PCR primer pairs for the amplification of the target sequence.","The outer primer pair amplifies a portion of the DNA molecule which includes the target sequence, and the inner primer pair amplifies part of the same portion of the DNA molecule that is amplified by the outer primer pair, but excludes the target sequence.","The inner and outer PCR primer pairs may comprise sequence tags.","The kit may further comprise secondary primers for the amplification of the sequence tags.","The present invention also provides a dideoxy DNA sequencing kit for producing short chain termination fragments.","The kit includes dNTPs and ddNTPs which are present in a low dNTP:ddNTP ratio."],["BACKGROUND OF THE INVENTION 1.Field of the Invention The invention generally relates to single nucleotide polymorphism (SNP) genotyping.","In particular, the invention provides a method for SNP genotyping based on a nested PCR design that creates structures directly suitable for both DNA sequencing and ligation reactions.","2.Background of the Invention With the completion of the rough draft of the human genome, over 1.5 million non-redundant SNPs have been identified and mapped by the publicly funded project and the SNP consortium (Sachidanandam et al, 2001).","The availability of such a large number of SNPs has prompted great interest in SNP related applications and research, such as large scale linkage, genomic association and pharmarcogenetic studies.","SNPs entered the central arena of genetics due to their abundance (Collins et al, 1997; Sachidanandam et al, 2001), stability (Sachidanandam et al, 2001) and the relative ease (Kwok 2000; Shi 2001; Syvanen 2002) with which they are genotyped.","The targets of SNP genotyping are normally small pieces of DNA, ranging from 150-300 base pairs.","Based on the SNP consortium report, the average SNP density in the human genome appears to be greater than 1 SNP/KB.","This high density provides a tool to analyze genomic structure at very high resolution, and to establish a direct correlation between genetic coding variation and biological function.","The relative stability of SNPs (compared to mini- and micro-satellite markers) in evolution makes it simpler to carry out this task.","Although SNP genotyping is generally easier than microsatellite genotyping, and despite the rapid and significant advances in SNP genotyping technology in the last few years, major issues remain unsolved.","Whereas only a few years ago, the main concern was the need to have a large number of SNPs available, currently the most urgent issues are increasing throughput and decreasing cost.","For example, for large scale applications as projected by Kruglyak (1999) and Long (1999), a reasonable cost would be <$0.10/genotype, or else large-scale projects will become prohibitively expensive.","To fulfill the goals of understanding the genetics of complex traits and common diseases, cost-effective and higher throughput methodology in SNP genotyping is essential.","Typically, genotyping protocols have three components: target amplification, allelic discrimination and product detection and identification.","They are normally executed sequentially, but can be processed in a single step reaction, depending on the means which are used for allele discrimination and signal detection.","Such single step procedures are well-suited to automation, but are not necessarily effective in terms of cost and throughput.","PCR is the dominant procedure utilized for target amplification.","As is widely known, a PCR can readily amplify targets present in lower copy number by a factor of 108 or more.","With respect to allele discrimination, all SNP genotyping technologies currently available are based on mechanisms involving one or more of the following: DNA polymerase, hybridization and DNA ligase.","DNA polymerase methods include single base extension (SBE) (Armstrong, 2000; Barta 2001; Bray 2001; Cai 2001; Chen 1997; Chem 1999; Chen 1998; Chen 1997; Chen 1997; Fan 2000; Lindblad-Toh 2000; Nikiforov 1994; Pastinen 1996; Pastinen 2000; Ross, 1998; Sauer 2000; Syvanen 1999; Ye 2001), de novo sequencing including pyrosequencing (Nordstrom 2000; Ronaghi 2001), allele-specific PCR and extension (Germer 1999; Myakishev 2001; Pastinen 2000), and structure-specific cleavage (Fors 2000).","Of these different forms, SBE is the most widely used and has been adapted for many different detection platforms.","Hybridization is also widely used in several different forms, including dynamic hybridization (Prince 2001), and is the primary method currently used in all microarray detection formats.","DNA ligase methods are based on the ability of DNA ligase to join the ends of two oligonucleotides annealed next to each other on a template (Tong 2000; Tong1999).","Two oligonucleotides can be designed to anneal to both sides of a SNP site, and by detecting the formation of ligation product, the genotype of a target can be inferred (Chen 1998; Delahunty 1996; Gerry 1999; Iannone 2000).","Once a target DNA sequence has been amplified and the allelic variants discriminated, the next step in a generic genotyping protocol is to detect and identify the allele specific products.","Detection mechanisms vary greatly, from simple fluorescence intensity (Armstrong 2000; Cai 2000; Chen 1998; Chen 1997; Delahunty 1996; Dubertret 2001; Fan 2000; Fergusoon 2000; Fors 2000; Germer 1999; Germer 2000; Gerry 1999; Iannone 2000; Lindblad-Toh 2000; Lindroos 2001; Marras 1999; Medintz 2001; Myakishev 2001; Nikiforov 1994; Pastinen 1997; Pastinen 2000; Prince 2001; Syvanen 1999; Ye 2001) to very precise mass (Bray 2001; Ross 1998; Sauer 2000) or electric charge (Gilles 1999; Woolley 2000) measurement.","The detection mechanisms roughly fall into two categories: homogeneous and solid phase mediated detection.","All homogeneous detection platforms depend on measuring fluorescence intensities and their change during and/or after reactions.","One common feature among homogeneous approaches is that they do not require any separation/purification prior to signal acquisition, making them amenable to automation.","Solid support techniques include flow cytometry genotyping (Armstrong 2000; Cai 2000; Ye 2001), zip-code microarrays (Fan 2000; Gerry 1999), and mass spectrometry genotyping (Bray 2001; Ross 1998; Sauer 2000).","Using a solid support in the detection step may potentially increase throughput and reduce cost, but unfortunately also entails the risk of complicating protocols and compromising data quality.","For example, when reaction mixtures are applied to a solid support, unintended binding of fluorescence dye to the solid surface can occur, necessitating extensive washing to minimize spurious signals.","With respect to cost, prices for genotyping technologies currently available on the market vary considerably, from $0.50-2.00/genotype.","For example, the template-directed dye-terminator incorporation assay with fluorescence detection (FP-TDI) (Chen 1999) costs roughly $0.50/genotype for reagents by list price.","For other methods, due to their requirement for special reagents and/or clean-up procedures, the cost is higher, e.g.","MALDI-TOF mass spectrometry is about $0.75-1.00, whereas pyrosequencing approaches $2.00/genotype.","No currently available technology approaches the low cost which is necessary in order to prevent large-scale projects from becoming prohibitively expensive, i.e.","<$0.10/genotype.","There are currently many genotyping applications in use or under development that require a relatively large number of both SNPs and samples (e.g., both in the thousands or more), demanding both cost effective and high throughput technologies.","Examples include gene mapping studies by linkage or linkage disequilibrium (Kruglyak 1999; Long 1999) for complex traits and pharmacogenetics (Riley 2000).","Clearly, these applications are critical to improving our understanding of the genetics of complex traits, common diseases and drug response, and to help attain the goal of individualized medicine.","However, none of the current approaches are suitable for such applications in terms of cost effectiveness coupled with high throughput potential, and SNP genotyping demands both.","SUMMARY OF THE INVENTION The present invention provides a methodology for high-throughput SNP genotyping that is highly cost effective.","This novel approach makes use of the high throughput capacity of DNA sequencers for SNP genotyping and is based on a nested PCR design that creates a series of ordered “structures” in parallel and exploits them for SNP genotyping assays.","The invention thus provides novel genotyping technologies that significantly increase throughput, reduce cost and are accessible for most researchers, leading to significant improvements in genotyping technology.","While typical SNP genotyping approaches currently available score <10 SNPs simultaneously, the methods of the present invention increase the number to about 50-100 with obvious advantages for throughput and cost.","The present invention provides a method of producing hybrid DNA with a single strand overhang that includes a target sequence.","The steps of the method include: obtaining a first primer which hybridizes to a 5′ strand of a strand of deoxyribonucleic acid (DNA), a second primer which hybridizes to a 3′ strand of the strand of DNA, and a third primer which hydridizes to said 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using the first primer, the second primer, and the third primer, an outer amplicon which includes a target sequence and an inner amplicon which excludes the target sequence; forming at least one of the following: a ligatable structure which includes a 3′-5′ sequence which excludes the target sequence hybridized to a 5′-3′ sequence which includes the target sequence, and a sequencible structure which includes a 5′-3′ sequence which excludes the target sequence hybridized to a 3′-5′ sequence which includes the target sequence; sequencing the sequencible structure(s) and ligating the ligatible structure(s) with a labeled oligonucleotide; and analyzing the sequencing products and ligation products with a DNA sequencer to determine the genotype of said individual.","The method may form both the ligatible structure and the sequencible structure.","The target sequence(s) may include a single nucleotide polymorphism.","The invention also provides a method of genotyping the deoxyribonucleic acid (DNA) of an individual by analyzing at least on target sequence in the DNA.","The steps of the method include: obtaining a first primer which hybridizes to a 5′ strand of a strand of said DNA, a second primer which hybridizes to a 3′ strand of the strand of DNA, and a third primer which hydridizes to the 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using the first primer, the second primer, and the third primer, an outer amplicon which includes said target sequence and an inner amplicon which excludes the target sequence; and forming at least one of the following: a ligatable structure which includes a 3′-5′ sequence which excludes the target sequence hybridized to a 5′-3′ sequence which includes the target sequence, and a sequencible structure which includes a 5′-3′ sequence which excludes the target sequence hybridized to a 3′-5′ sequence which includes the target sequence; sequencing the sequencible structure and ligating the ligatible structure with a labeled oligonucleotide; and analyzing the sequencing and ligation products so produced with a DNA sequencer to determine the genotype of the individual.","The forming step of the method may form both the ligatible structure and the sequencible structure.","The target sequence(s) may include a single nucleotide polymorphism.","The sequencible structure may be sequenced by a technique such as dideoxy sequencing, pyrosequencing, and single base extension.","In a preferred embodiment, of the method, i) a plurality of target sequences is analyzed; ii) the step of forming forms an ordered series of sequencible structures; iii) the step of sequencing produces an ordered series of sequencing products of varying, non-overlapping length; and iv) the step of analyzing is carried out by electrophoresing the ordered series of sequencing products in a single channel of said DNA sequencer.","In some embodiments, the step of sequencing is carried out by single base extension (SBE).","In other embodiments, the step of sequencing is carried out by a dideoxy sequencing reaction utilizing a ratio of dNTPs to ddNTPs that is lower than that which is typically used in order to produce a short sequence reading.","In a preferred embodiment of the present invention, the labeled oligonucleotide is fluorescently labeled.","In one embodiment of the method, i) a plurality of target sequences is analyzed; ii) the labeled oligonucleotides are degenerate; iii) the ligation products are of varying, non-overlapping lengths; and iv) the step of analyzing is carried out by electrophoresing a plurality of ligation products in a single channel of said DNA sequencer.","The present invention also provides a method for analyzing at least one target site in a DNA molecule.","The method includes the steps of: 1) amplifying the target site(s) by nested PCR; (The nested PCR is carried out using inner and outer PCR primer pairs, wherein the outer PCR primer pair forms a first PCR product which contains a target site, and the inner PCR primer pair forms a second PCR product which contains a portion of the first PCR product but does not contain the target site); 2) denaturing the first and said second PCR products to form ssDNA sequences, 3) reannealing the ss DNA sequences to form a sequencible hybrid DNA molecule and a ligatible hybrid DNA molecule, 4) performing sequencing reactions with said sequencible hybrid DNA molecule and ligation reactions with said ligatible hybrid DNA molecule, and 5) determining the characteristics of the target sequence by analyzing results obtained in the performing step.","In a preferred embodiment of the method, the target site(s) may be SNP polymorphism sites.","Further, the inner and outer primer pairs may comprise a sequence tag, which in turn may comprises a restriction enzyme recognition site.","In some embodiments of the method, the step of amplifying may be carried out using a low concentration of primers, and may further comprising a second step of amplification using secondary primers for amplification of the sequence tags.","According to the method, one or several target sites may be analyzed.","The step of amplifying may be carried out in a single multiplex PCR reaction, or in multiple independent PCR reactions, and the results may be analyzed by a DNA sequencer.","The invention also provides inner and outer PCR primer pairs for the amplification of a target sequence in a DNA molecule, wherein the outer PCR primer pair forms a first PCR product which contains the target sequence, and the inner PCR primer pair forms a second PCR product which contains a portion of the first PCR product but does not contain said target sequence.","The target sequence may be an SNP polymorphism site.","The primer pairs may comprise a sequence tag.","The present invention also provides a kit for amplification of at least one target sequence in a DNA molecule.","The kit includes inner and outer PCR primer pairs for the amplification of the target sequence.","The outer primer pair amplifies a portion of the DNA molecule which includes the target sequence, and the inner primer pair amplifies part of the same portion of the DNA molecule that is amplified by the outer primer pair, but excludes the target sequence.","The inner and outer PCR primer pairs may comprise sequence tags.","The kit may further comprise secondary primers for the amplification of the sequence tags.","The present invention also provides a dideoxy DNA sequencing kit for producing short chain termination fragments.","The kit includes dNTPs and ddNTPs which are present in a low dNTP:ddNTP ratio.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1.A schematic drawing illustrates a nested PCR design that would create both a sequencible and a ligatible structure.","The reaction uses two primer sets (inner and outer).","P1, forward outer and forward inner primer; P2, reverse inner primer; P3, reverse outer primer.","When this design is multiplexed, multiple sequencible and a ligatible structures are formed.","These structures can be used for genotyping reactions.","FIGS.","2A and B.","A, a multiplex PCR scheme that amplifies three target sites.","Two primer sets (inner and outer) are used for the amplification of each target site.","The target sites are represented by “x”.","All primers contain two domains, a common domain and a target specific domain.","The target specific domains, indicated by arrows, have sequences unique to the targets, whereas the common domains are not target specific.","=common domain of forward inner and forward outer primers; ▮=common domain of inner reverse primers, - - - =common domain of reverse outer primers.","B, close-up view of common domain of inner primer shows restriction sites incorporated into the sequence of the domain.","FIGS.","3A and B.","A. Depiction of nested PCR primer design.","The sequence (SEQ ID NO.","1) represents 393 nucleotides from human chromosome 6.Nucleotides corresponding to primer pairs are shaded with gray, and arrows indicate primer orientation.","Arrow indicates site of polymorphism.","The sequence which was generated by ABI 377 sequencer is shown in italics, and is also depicted in FIG.","3B (SEQ ID NO.","2).","The sample is a A/A homozygote.","FIGS.","4A and B.","A. Depiction of nested PCR primer design.","The sequence (SEQ ID NO.","3) represents 494 nucleotides from human chromosome 6.Nucleotides corresponding to primer pairs are shaded with gray, and arrows indicate primer orientation.","Arrow indicates site of polymorphism.","The sequence which was generated by ABI 377 sequencer is shown in italics, and is also depicted in FIG.","4B (SEQ ID NO.","4).","Two samples were shown in FIG.","4B.","The upper panel was a heterozygote (C/T), at base position 24, as pointed by the arrow, both bases © and T) were identified.","The lower panel was a homozygote (T/T), at the indicated position only the T base was identified.","The example demonstrated that a single nucleotide polymorphism (SNP) could be identified by sequencing from the sequencible structure formed from the nPCR design.","FIG.","5.SBE genotyping using sequencible structures from nPCR products amplified by Pfu DNA polymerases.","Nested PCR primers were designed for an SNP marker from human chromosome 6.The inner product size is 189 bp, its 3′ end is positioned immediately upstream to the polymorphic site when it forms sequencible structures.","PCRs were performed with DNA samples of known genotypes.","After PCRs the outer and inner products were purified and denatured together to form sequencible structures.","SBE reactions were performed and products analyzed in ABI 377 Sequencer using Genescan.","Three samples were presented.","The upper panel is a homozygous A/A where an SBE product of 190 bp is found (arrow).","Products of the same size are found in the heterozygous (middle panel) and the other homozygous (lower panel).","The peaks after the dye front are ROX500 size makers (35, 50, 75, 100, 139, 150, 160, 200, 250 bp respectively).","FIG.","6.An example illustrates a short sequence read with a specially assembled sequencing mixture that has a different ddNTPs/dNTPs ratio.","The components used for the example were the following: 1.75 μM of −21 M13 primer, 300 ng of pGEM, 60 μM of each dNTPs, 2.5 μM of R6G-acyclo-ATP, 12 μM of ROX-acyclo-CTP, 10 μM of BODIPY-Fluorescein-acyclo-GTP, 25 μM of TAMRA-acyclo-UTP, 1 unit of AcycloPol DNA polymerase and 1× AcycloPol reaction buffer.","The thermal cycling conditions were 96° C. for 3 min., followed by 25 cycles of 96° C. for 1 min., 50° C. for 15 sec.","and 60° C. for 4 min.","FIG.","7.Primer designs affected multiplex PCR and nPCR.","Lane 1 was a 4×multiplex PCR with simple PCR design, Lane 2 was the same 4 amplicons but with the two domain design, and Lane 3 was the 4× multiplex nPCR with the same amplicons performed in two sequential reactions.","In the first reaction, equal amount of primary primers at 10 nM were used.","The cycling condition was 95° C. for 10 min, followed by 10 cycles of 95° C. for 45 sec., 65° C. for 5 sec., ramping to 55° C. at 0.1° C./sec and 55° C. for 90 sec.","For the second reaction the primers used were different among the three lanes.","Lane 1, since it did not have secondary primers, the same primers were used as in first reaction but the concentration increased to 100 nM.","Lane 2 used two secondary primers, M13 forward and reverse, each 400 nM.","Lane 3 used three primers, M13 forward (600 nM), M13 reverse (400 nM) and the common tag for inner PCR primers (400 nM).","The cycling conditions for the second reaction was following: 95° C. for 10 min, followed by 30 cycles of 95° C. for 45 sec., 55° C. for 90 sec.","DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION The present invention provides methods of high-throughput SNP genotyping that are highly cost-effective.","The methods utilize the high-throughput capability of ubiquitous and readily accessible DNA sequencers in order to carry out the SNP analysis.","This is possible due to the experimental design of the methods, which are based on nested PCR reactions.","In a preferred embodiment, the target sites which are analyzed are SNP polymorphism sites.","However, those of skill in the art will recognize that other target sites may also be analyzed by the methods of the present invention.","For example, short insertions and deletions, and closely spaced multiple SNPs can be identified by the multiple target sequencing approaches disclosed herein.","As is well-known to those of skill in the art, nested PCR is a PCR amplification method that “nests” one PCR reaction within another.","Two sets of primers are used: “outer” primers amplify a portion of a DNA strand, and “inner” primers amplify a smaller portion of the same DNA that is located within the larger, outer portion.","Thus, two PCR products (amplicons) are produced.","One amplicon is smaller than the other and contains a “subset” of the bases contained in the larger, outer amplicon.","In the practice of the present invention, a new design for nested PCR is used.","Primers are designed so that the outer amplicon includes a target site (e.g.","an SNP polymorphism site) and the inner amplicon excludes the target site.","The purpose of the inner PCR in the nested reaction is to synthesize a DNA fragment whose size and position relative to the outer PCR can be precisely controlled.","This design concept is illustrated in FIG.","1.As can be seen, in FIG.","1, the inner primer set is composed of primers P1 and P2, and the outer primer set is composed of primers P1 and P3.","(In this example, Primer P1 is utilized in both primer sets.)","The P1-P3 outer primer set flanks the indicated target site whereas the P1-P2 inner primer pair does not.","PCR amplification of the DNA with these primers will produce two amplicons.","The larger of the two is the outer amplicon formed by amplification with the P1-P3 pair; this amplicon includes the target site.","The smaller of the two amplicons, the inner amplicon, contains base pairs which lie within the bounds of the larger outer amplicon (i.e.","is “nested” within the larger amplicon), but does not contain the target site.","The ultimate product of the PCR reaction is double strand DNA of two lengths, only one of which (the longer) contains the target site, and the shorter of which contains a subset of the bases contained in the longer strand.","In a preferred embodiment of the invention, the same primer functions as both the outer and inner forward primer.","However, those of skill in the art will recognize that this need not be the case.","With reference to FIG.","1, a distinct forward inner primer (“P4”, shown in parentheses) might be designed to anneal between P1 and P2 and in the orientation of primer P1.Because the shorter amplicon contains bases which are identical to a portion of the longer amplicon, when the ds strands are denatured and allowed to reanneal, some (approximately 50%) of the shorter single strand DNA of the inner amplicon will hybridize with matching portions of the single strand DNA of the outer amplicon.","This cross hybridization between the two amplicons creates two different inner-outer “hybrid” structures as depicted in FIG.","1.In one of the two structures, the “ligatible structure” of FIG.","1, a 3′ overhang will be present, the bases of which originated from the outer amplicon and contain the target site.","This structure can be used, for example, for ligation based applications.","By “ligation based applications” we mean the smaller amplicon is one of the two or more pieces of DNA that are joined together by a DNA/RNA ligase.","In the other structure, the “sequencible structure” of FIG.","1, a 5′ overhang will be present, the bases of which originated from the outer amplicon and also contain the target site.","This second type of structure will be directly amenable to sequencing applications.","By “sequencing applications” we mean that the smaller amplicon acts as a sequencing primer to sequence the DNA downstream from it.","The sequencing can be only one base in length (i.e.","SBE) or in the tens or in the hundreds of bases.","Thus, in a single nested PCR reaction followed by denaturation and annealing, it is possible to generate both sequencible and ligatible structures.","Of course, some (approximately 50%) of the single strand DNA will renature to form the original ds PCR products (“structures same as products” in FIG.","1).","Years of improvement in primer design and thermocycling conditions have increased the success rate for single target PCR reactions to greater than 90%.","However, multiplex reactions such as those in a nPCR reaction, have a much lower success rate and extensive optimization is typically required, usually because it is very difficult to obtain amplified products in equal amounts.","The usual solution is to increase the primer concentration for the weakly amplified target(s).","This suggests that it is annealing efficiency that is responsible for the unequal amplification.","In U.S. Pat.","No.","5,882,856 to Shuber, the complete contents of which is herein incorporated by reference, an alternative solution is proposed in which all primers utilized have two domains, one which is target specific as in regular PCR designs, and one which is a “common domain”.","For a set of PCR primers to be multiplexed, two common domains are utilized.","A first common domain is a sequence shared by all forward primers and is attached at the 5′ end of the target specific domain.","A second common domain sequence is shared by all reverse primers and is attached at the 5′ end of the target specific domain.","The common domain facilitates annealing of different primer sets because the common domain has a higher annealing temperature than the target specific domain.","In the practice of the present invention, common sequence domains or “sequence tags” as taught by Shuber may be utilized for both inner and/or outer PCR primers.","The common sequence tag for the outer PCR primers would help in multiplexing PCR so that amplification would result in the production of equal amounts of product.","The common sequence tag for the inner primers also serves to even the levels of PCR products, and in addition may provide restriction sites that can be used to generate precise fragment ends for genotyping reactions such as sequencing and/or ligation reactions.","The combination of incorporating special restriction enzyme recognition sequences in such a sequence tag, and the deliberate positioning of the inner PCR primers make it possible to precisely tailor the resulting PCR products for any of a variety of purposes, including but not limited to primer extension, sequencing, ligation, pyrosequencing, structure specific cleavage, and other genotyping reactions.","Those of skill in the art will recognize that placement of the P2 primer can be anywhere between the target site and the other primer of the inner primer pair, so long as a useful PCR product is produced by PCR amplification of the inner primer pair, and the target site itself is not amplified.","The positioning of the P2 primer relative to the target SNP may vary depending on the mechanism of the genotyping reactions.","For example, for SBE reactions, the end of the primer should be immediately 5′ to the SNP site.","For Multi-Target Sequencing (NulTarSeq) reactions, there may be a gap between the end and the target SNP site.","In a preferred embodiment, for MulTarSeq reactions the gap may be from about 1 to 50 bases, or more preferably from about 5 to about 20 bases, and most preferably from about 10 to about 15 bases.","Further, as described above, the desired length of the final product from the amplification of PCR primers containing common sequence tags may also be determined by the presence of a restriction enzyme cleavage site within the tag.","In yet another embodiment of the present invention, the amplification reactions may be carried out in a step-wise fashion using primary and secondary primers.","In this embodiment, a single PCR is divided into two sequential reactions, the first of which uses the primary primer set and the second of which uses the secondary primer set.","The primary primers may be designed to have two domains (target specific and common) as described above.","The secondary primer set contains only the common tails from the primary primers.","For the first reaction, the primary primers are used in only limited but equal amounts, e.g.","about 1-5% of the amount for a regular PCR i.e.","about 0.1 to about 100 nM.","The rationale is that by limiting the amount of primers for each set multiplexed in the reaction, the system is forced to produce limited but equal amounts of products for all amplicons.","When the more robust primers are used up (because of the limited amount available) the system will work with the less efficient primers.","The purpose of the primary PCR is to amplify enough templates from the genomic DNA to carry out the secondary PCR.","Since all primary primers have limited but equal amounts, this forces the primary PCR to produce limited but near equal amounts of templates for the secondary PCR.","For the secondary PCR, the secondary primers, consisting of only one set for all primary amplicons, is used.","In essence this secondary reaction is a single PCR that amplifies multiple templates.","The net result of this two-step PCR process is to compensate for differences in efficiency across PCR reactions.","The purpose of the inner PCR in the nested reaction is to synthesize a DNA fragment whose size and position relative to the outer PCR can be precisely controlled.","By altering the position of the P2 primer relative to P1, and by optionally including sequence tags with restriction sites, it is possible to design longer or shorter PCR fragments.","The synthesized fragments can then be used in any of a variety of genotyping reactions, including but not limited to primer extension, sequencing, ligation, pyrosequencing, structure specific cleavage, and the like.","For example, the sequencible structure fragments may be used as extension primers in SBE and MulTarSeq reactions.","The direct production of sequencible structures by the methods of the present invention thus permits investigators to take advantage of the ready availability of DNA sequencers for the purposes of genotyping.","Relatively long primers can be synthesized in this manner in a feasible and much more cost effective way than, as is typically done, by chemical synthesis.","For example, the length of such a fragment can range from about 35 to about 1000 bps, and is preferably in the range of about 50 to about 650 bps.","To use a standard sequencing reaction to determine the sequences surrounding a polymorphic site, it is simply necessary to add an appropriate sequencing mixture without using any sequencing primer, because the sequencible structure provides the free 3′ end for the sequence reaction.","Those of skill in the art will recognize that several methods of sequencing are well established, and that new sequencing methodologies are continually being developed.","In the practice of the present invention, any suitable method of sequencing may be utilized.","Examples of such methods include but are not limited to Sanger (dideoxy)-based sequencing methods and pyrosequencing.","The procedure illustrated in FIG.","1 amplifies a single target and relies on multiplex PCR amplification since more than one pair of primers is used in a single reaction.","However, those of skill in the art will recognize that in many cases it will be desirable to amplify more than one target site at a time.","In this case, the design of the inner primer pairs for each target site may be such that an ordered series of PCR products of varying lengths is produced, i.e.","the sequencible structure formed for each target site differs in length from the sequencible structures formed for each of the other target sites.","(See FIG.","2A).","For example, an ordered series of sequencible structures from about 50 to about 500 bps in length can be designed, thus adding an additional distinguishing feature to each of the amplified target sites.","Because of this feature, multiple sets of PCR products can be sequenced in a single reaction providing that the length of sequencing is limited so that their chain termination fragments do not overlap in a sequencing gel.","This would allow the sequencing of multiple targets in a single reaction and analysis in a single channel (e.g.","a lane or capillary).","Those of skill in the art will recognize that several ways exist to control the length of reading for sequencing reactions.","For example, the length of reading can be adjusted by changing the ratio of ddNTP/dNTP in the reaction mixture.","For standard Sanger sequencing reactions, the ratio is typically about 1:100, a ratio intended to promote relatively long reads (e.g.","about 600-800 bps).","Those of skill in the art will recognize that, by systematically altering and testing the ratio, it is possible to find the optimal reading length for a given application.","For most SNP genotyping purposes, a read of from about 1 to about 50, and preferably from about 1 to about 25 bases would be sufficient, requiring a low ddNTP/dNTP ratio of preferably in the range of about 1:0 to about 1:10, and more preferably about 1:0 to about 1:5.For other applications, the optimal length can be attained by adjusting the ratio, i.e.","the ratio may be optimized by methods which are well known to those of skill in the art in order to achieve the desired sequence read length.","Precise control of the read allows very high level multiplexes to be carried out (e.g.","100 to 200×).","In order to carry out the practice of the present invention, primer design involving placement of the primer and the optional addition of a sequence tag must be carried out.","Further, those of skill in the art will recognize that other factors influence primer design and must be taken into account as well.","For example, the exact sequence of bases will influence the strength of the hybridization of the primer with its cognate DNA (the sequences of based to which it binds during PCR) and thus must be taken into account when planning PCR amplification protocols; the possibility of primer-dimer formation is taken into account, as is the potential for the formation of secondary structure, etc.","Those of skill in the art are well acquainted with such issues and numerous resources exist to aid the skilled practitioner in these various aspects of primer design.","As discussed above, the common domains of the inner primers may be designed to include restriction enzyme recognition sites.","The restriction enzymes preferred are those that cleave DNA strands downstream from their recognition sites and generate blunt ends products.","For sequencing applications restriction enzymes that cleave DNA downstream and generate 3′ overhangs can also be used.","For ligation applications restriction enzymes that cleave DNA downstream and generate 5′ overhangs can also be used.","Examples of such sites include but are not limited to these enzymes: Mly I, Fst F51, Bpm I, Eco571, Bsg I, Eci I, Bst71 I, Alw26 I, Ksp62 I, BceA I, etc.","The other nucleotides in the sequence of a common domain inner primer are selected to balance the melting temperature with that of the common domain of the outer primers.","With respect to the design of common domain sequences for outer primers (i.e.","those which do not contain restriction sites), the sequences are selected so that so it will match the melting temperature of the common domain of the inner primer.","In general, the generation of the sequencible and ligatible structures of the present invention will be carried out as follows: a PCR amplification reaction is carried out, followed by a restriction with a suitable restriction enzyme if a restriction site was incorporated into a common sequence tag of the inner primers.","This is followed by treatment to “clean up” the reaction e.g.","with shrimp alkaline phosphatase (SAP) and exonuclease I (Exo I) to degrade excess PCR primers and dNTP left over from the PCR reaction, and to remove the overhanging ends of restriction digestion.","(Alternatively, filtration via sephadex gel may also be employed.)","After SAP/Exo I digestion, a high temperature denature step is performed.","This denature step serves two purposes: one is to inactivate both SAP, Exo I and the restriction enzyme; the other is to completely denature double stranded DNA.","During the cooling process that follows, the four single stranded DNA fragments from the two PCR products will form four kinds of structure.","Two of them are the exactly same as the PCR products and are both blunt ended.","Two new structures formed between the complementary strands of the large and small PCR products, one of them with a 5′ end single stranded, the other one with a 3′ end single stranded.","The one with a 5′ end single stranded provides a free 3′ end for sequencing reaction/primer extension and for ligation, and is referred to as a “sequencible structure”.","The one with single stranded 3′ end provides a free 5′ end for ligation, and is referred to as a “ligatible structure”.","These four structures will have equal concentrations if the quantity of the two PCR products is equal.","In this case, the sequencible and ligatible structures will represent about 50% of the reannealed products.","This amount is sufficient for both sequencing/primer extension and ligation reactions to make new products.","In some embodiments of the present invention, a single target site is analyzed by the methods of the present invention, and the PCR step is carried out by multiplex PCR, i.e.","amplification with both the inner and outer primers is carried out in the same reaction.","However, those of skill in the art will recognize that two separate PCR reactions may also be carried out, i.e.","the inner and outer PCR reactions are carried out separately.","In this case, the PCR products from the two reactions may be combined later at some point prior to the renaturation step so that the single strands can reanneal.","Similarly, in other embodiments of the present invention, it may be desirable to PCR amplify more than one target site at a time.","In some cases, this may be carried out by a multiplex PCR reaction in which inner and outer primers for all target sites of interest are amplified simultaneously in a single reaction.","Alternatively, one or more sites may be amplified together, or each site may be amplified alone, either in a single reaction with both inner and outer primers, or in two reactions, one for the inner and one for the outer primers as described above.","Any combination of PCR amplification reactions may be utilized in the practice of the present invention, so long as the procedure results in the production of suitable sequencible and/or ligatible structures.","In a preferred embodiment, the present invention provides methods which ultimately utilize DNA sequencers or the like to detect the products formed from sequencible and/or ligatible structures created by the present methods.","Those of skill in the art will recognize that many suitable techniques exist by which the sequencible products can be produced for analysis by a DNA sequencer.","Examples include but are not limited to Sanger-type sequencing reactions, single base extension (SBE), and pyrosequencing.","Ligatible structures may be analyzed directly on a DNA sequencer after a labeled oligonucleotide (e.g.","fluorescently labeled) is joined to the 5′ phosphate of the ligatible structure via a ligation reaction.","Those of skill in the art are well acquainted with Sanger-type sequencing reactions, which are based on the interruption of enzymatic extension of the DNA chain at the 3′ hydroxyl via incorporation of 2′-3′-dideoxy analogs.","Four sets (one for each dideoxy analog) of such chain-terminated fragments of differing lengths are generated and “read” (detected) by a DNA sequencer to yield a sequence.","Typically, the fragments are labeled with radioactivity or fluorescent tags for detection.","Single base extension is essentially a sequencing reaction, the difference being that SBE uses only dye-terminators, resulting in the extension of the primer by only a single base.","As described above, in the practice of the present invention, the length and position of the extension primer relative to the polymorphic site may be controlled so that it is possible to create a primer which anneals immediately upstream of a polymorphic site.","Then, in a regular SBE reaction, the polymorphic base can be determined by identifying which base(s) is incorporated into the primer.","If several target sites are to be analyzed, a set of primers can be designed which, upon amplification of the targeted DNA, produce an ordered set of such sequencible structures of varying, non-overlapping lengths.","When such structures are extended by dye-labeled ddNTPs, an ordered series of SBE products of varying, non-overlapping lengths is produced.","In the proposed nested PCR design, because extension primers from about 50 to over 500 bases can be obtained, it is possible to separate these SBE products in a sequencing gel/capillary.","A regular sequencing gel can sequence about 500-800 bases.","Therefore, over 100 SNPs can be analyzed in a single channel by loading their corresponding SBE products in one lane/capillary when the extension primers are designed to be positioned, for example, every 4-5 bases apart for a full range of 500-600 bases.","In order to successfully carry out SBE reactions, the ends of the primers must be precise, i.e.","it is necessary to control precisely where the inner amplicon ends.","The use of Taq DNA polymerase may in some cases be disadvantageous due to its known extendase activity which interferes with such precision.","Those of skill in the art will recognize that a variety of means are available to overcome such a problem.","For example, the use of high fidelity polymerases may be employed.","Pfu DNA polymerase is a well characterized DNA polymerase that has been shown to have high fidelity of replication (Cline 1996).","It is known that Pfu DNA polymerase does not add extra A's at the 3′ end (Cline 1996; Hu 1993).","Based on a study by Cline et al (Cline 1996) the use of Pfu DNA polymerase gives precise ends of inner PCR products.","A second alternative is to use ribonucleotides and/or abasic analogs in the inner primer to control DNA polymerases' end point.","Some DNA polymerases do not have reverse transcriptase activity, Pfu and Vent polymerases are examples (Perler 1996).","These enzymes can not incorporate any nucleotide when they encounter a ribonucleotide base in a template.","Coljee et al recently reported a strategy to create precise overhang at the ends of PCR products by using ribonucleotides (Coljee 2000).","The strategy inspired us to use same mechanism to control the ends of inner PCR products.","Thus, inner primers may be designed to overlap the target SNP site, at the SNP site, instead of using regular deoxyribonucleotide, a ribonucleotide is used.","DNA polymerases can use a chimeric oligonucleotide as primer efficiently (Gal 2000; Stump 1999), so the use of chimeric primers would not compromise PCR efficiency.","When the complementary strands are synthesized, the DNA polymerases will stop one base before the ribonucleotide base in the chimeric inner primers, creating very accurate and precise ends for SBE.","It is also possible to create chimeric primers containing several ribonucleotide bases to create several base overhang for the purpose of ligation (Coljee 2000).","Similarly, a nucleotide analog that does not contain the base, the abasic analog, has similar effects on DNA polymerase (Gal 2000; Gal 1999) and may be used.","Yet another alternative is to use restriction enzymes to produce precise 3′ ends for sequencible structures.","There are some restriction enzymes that cleave DNA strands downstream from their recognition sites, e.g.","KSP632 I, Alw26 I, and Bst71 I.","As discussed above, to take advantage of these features, a common domain (sequence tag) may be added to the 5′ end of the inner PCR primers.","The common domains contain restriction enzyme sites which can be positioned so that the 3′ extending ends are located just before the polymorphic sites after the tail is cleaved off.","The use of a common domain for inner primers could also help to even the amounts among different amplicons when multiplexed.","Pyrosequencing is a unique technique that incorporates a sequencing reaction and real time detection together.","Since, as in a regular sequencing reaction, pyrosequencing uses DNA polymerase, the free 3′ termini in the sequencible structures can function as extension primers.","Since pyrosequencing is a real time system, it is not amenable to the analysis of multiplex reactions.","There would thus be no need to use a common tail for the inner PCR primers.","The primers for a reaction whose products are destined for analysis by pyrosequencing are designed to accommodate the needs of pyrosequencing, preferably from about 5 to about 8 bases upstream of the target polymorphic site.","The ligatible structures produced by the methods of the present invention have a free 5′ phosphate.","If a common tag is included in the design, when the common tag is removed from the inner PCR products, restriction enzyme digestion leaves the 5′ phosphate on the ligatible structure.","To make such a ligation site at a polymorphic site, the cleavage site of restriction enzymes used in the inner PCR primer must be immediately down stream of the polymorphic site on the complementary strand.","Those of skill in the art will recognize that, because all of these restriction enzymes produce a 5′ over hanging strand, a suitable design of restriction site for ligation assay would not be suitable for SBE reaction, but could be used for MulTarSeq reaction.","Further, one advantage of the use of ligation to score a polymorphism is that it can be combined directly with the PCR reaction; resulting in a closed tube assay.","To be detected by a DNA sequencer, DNA fragments have to be labeled with fluorescence dyes.","In the present nested PCR design, the PCR products are not labeled with any fluorescence dye.","Thus, the ligatible structures can be detected only if they are ligated to a labeled oligonucleotide, e.g.","one that bears a fluorescent label.","To be ligated onto a ligatible structure created by restriction enzyme digestion and subsequently detected by a DNA sequencer, an oligonucleotide would have a 3′ OH group and 5′ fluorescence labeling.","Current DNA sequencers typically analyze fluorescence with a spectrum between 510-610 nm.","Those of skill in the art will recognize that there are many fluorescence dyes available in this range.","For example, the BODIPY series of fluorescent dyes have been shown to minimize emission overlaps between dyes (Metzker 1996).","Examples of other dyes which may be utilized to carry out this aspect of the invention include but are not limited to FAM, fluorescein, R110, R6G, TAMRA, ROX, Texas red, etc.","If only a single locus is of interest is to be analyzed, locus-specific fluorescence-labeled oligos may be utilized.","However, in order to carry out high level multiplex reactions, sets of degenerate, fluorescence-labeled oligos are desirable.","For example, in order to make ligatible structures suitable for high throughput genotyping assays, a common set of four fluorescence dye labeled degenerate oligonucleotides may be utilized.","The degeneration of the oligonucleotides is to make them reusable for all potential assays.","If an oligonucleotide with a specific sequence is used, it would only work for one particular target.","Several important issues must be taken into consideration regarding the design of the degenerate oligonucleotides.","One is the length of the oligonucleotide, which effects both ligation efficiency and the concentration of a specific sequence in the degenerate oligo population).","For example, for a 6-7 mer oligo containing 5-6 degenerate bases, any given sequence would have a concentration of ¼3-¼4 (i.e.","1/1024- 1/4096).","If the total concentration for a 7-mer degenerated oligo used in ligation is 1 μM, the effective concentration for any giving sequence would be greater than 0.2 nM, a concentration at least 100 fold above the detection limit of sequencing gel/capillary.","Those of skill in the art will recognize that the optimal length of the degenerated oligo and its concentration may vary from experiment to experiment, depending on the level of degeneracy and multiplex used in the reaction.","However, the oligo will generally be in the range of about 5- about 20 bps in length with from about 4 to about 19 degenerate bases, and more preferably in the range of about 6 to about 8 bps in length with about 4 to about 6 degenerate bases.","Another consideration is the melting temperature for the degenerated oligonucleotide.","As is well known to those of skill in the art, the Tm will vary depending on factors such as the GC content, the length of the oligo, and the like.","For example, increasing the GC content and/or the length of an oligo increases its Tm.","Increasing the length of oligonucleotide will increase Tm, but would decrease the effective concentration for a specific sequence.","In addition, other factors such as the ionic strength of the ligation buffer also affect Tm significantly.","Other considerations involved in oligo design which are well-known to those of skill in the art include the number and position of bases which are degenerated.","The number of degenerated bases will have a significant impact on the effective concentration and Tm.","For example, if 5 degenerated bases are used within a 7 mer, 16 oligos must be synthesized, but if 6 degenerated bases are used, only 4 oligos are necessary.","While one specified position must match the polymorphic site if multiple specified positions are used, the position does not necessarily have to be the last one on the 3′ end.","Further, the position of the ligation site relative to SNP site (the 5′ end or the 3′ end) may also be altered to optimize a given experiment.","Previous studies (Tong 2000; Tong 1999) have suggested that DNA ligase exhibits better allelic discrimination when the ligation site is on the 3′ end.)","Those of skill in the art will recognize that many tools exist and are well-known to the skilled practitioner for optimizing such parameters, thereby facilitating the design of suitable oligos.","(See, for example, Delahuntyl996, Tong 2000; Tong 1999).","Examples include but are not limited to various computer soft ware packages such as DNA Star, Oligo, Primer Express, etc.","Other factors which may be considered include the type and concentration of ligase to be employed.","For example, a 6-7 mer oligo with a moderate GC content should exhibit a melting temperature between 3040° C., a Tm sufficient to perform a cyclic ligation assay.","The use of thermal stable DNA ligase would provide an opportunity for multiple cycling of a ligation assay at this temperature, which would compensate for lowered efficiency of the reaction.","Those of skill in the art will recognize that a plethora of protocol references, kits, etc.","are readily available to facilitate carrying out such manipulations of the DNA.","The precise nature of the protocols which are used is not a central feature of the present invention, and any of many suitable means for carrying out the various steps of PCR amplification and detection and analysis of the amplicons may be utilized in the practice of the present invention.","Those of skill in the art will recognize that a variety of additional well-known techniques may be reliably utilized in the practice of the present invention, examples of which include but are not limited to: mass spectroscopy, ELISA techniques, fluorescence intensity/energy transfer/polarization, various micro assays (e.g.","DNA/RNA arrays), microfluidic devices, and magnetic/color-coded micro beads.","In a preferred embodiment of the present invention, the method of detection is capillary electrophoresis.","The demand from the human genome project greatly stimulated technology development in DNA sequencing and, in return, improvements in DNA sequencing greatly speeded up the human genome project.","As a result, capillary DNA sequencers became widely available to researchers in both academe and industry.","Capillary DNA sequencers are highly sensitive and automated detection systems.","They also have a relatively broad detection range, covering at least three orders of magnitude.","Their analytical power is better than most signal detection systems currently used for SNP genotyping.","Capillary DNA sequencing systems have been proven to deliver ultra high throughput in many DNA sequencing centers around the world.","Capillary electrophoresis has been used in DNA polymorphism analysis (Barta, 2001; Medintz 2001).","Most of these studies exploited the sensitivity and speed of separation.","Lindblad-Toh et al (2000) used a DNA sequencer to increase throughput.","In this study, the authors chemically synthesized oligonucleotides from 18-50 bases as extension primers and succeeded in scoring 14 SNPs in one lane (The size difference between primers was 4 bases, there were a few sets of oligos of the same length but of different sequences that were used to score two different SNPs by different colors).","This study essentially proved that DNA sequencers had the potential for high capacity SNP scoring using the SBE format, and the length of primers was the limiting factor for throughput.","The wide availability, high detection sensitivity, high throughput capacity and automation readiness of capillary DNA sequencers call for high capacity applications, and SNP genotyping is one of such applications.","If these two are united, considerable advances in cost and throughput would be achieved.","Many did not consider DNA sequencers for high throughput SNP scoring mainly because it was not cost-effective.","DNA sequencers separate DNA fragments by sizes and colors.","In order to separate genotyping products by a sequencer, the products must be different in sizes and or colors.","To separate two SBE products, for example, SBE primers must be different in length or the extended bases must be different in color.","Otherwise, the two products of same length and same colors would not be distinguished.","While it is reasonable to chemically synthesize primers of 15-40 bases, this size range gives only limited multiplex capacity (Lindblad-Toh 2000)).","To chemically synthesize primers longer than 50 bases is not economical, especially in high throughput settings.","But a sequencer can separate DNA fragments up to 600-800 bases.","In the practice of the present invention as described above, SBE products can be arranged to be 5 bases apart for 600-800 bases, allowing the loading over 100 SBE products in one lane.","Multiplexing at this level makes it reasonable to use sequencers for SNP scoring.","Furthermore, practical protocols for using DNA sequencers are well-established and many handling and procedures involved have been automated and are readily available for large scale applications.","The following examples are intended to be illustrative in nature and are not intended to be limit the scope of the invention in any way.","EXAMPLES Example 1 Formation of Sequencible Structures Sets of nPCR primers were designed as described to amplify DNA fragments from six markers of human chromosome 6 and individual nPCR was performed using Taq-Gold DNA polymerases.","Clean-up digestion was carried out using shrimp alkaline phosphatase and E. coli exonuclease I (37° C. 1 hour) After the clean-up digestion, a high temperature denaturing (95° C. for 15 min.)","was performed and the samples were allowed to cool to room temperature.","A sequencing reaction was performed using standard sequencing kits but without sequencing primers.","Sequencing reactions were performed either: 1) under standard cyclic sequencing program: 25 cycles of 96° C. for 1 min., 50° C. for 15 sec.","and 60° C. for 4 min; or 2) using this condition: 25 cycles of 96° C. for 1 min., 80° C. for 4 min.","The purpose of these experiments was to test whether a sequencible structure formed after denaturing and reannealing of the outer and inner PCR products together.","If we obtained sequences and the sequences matched the inner PCR primer position, this would confirm that sequencible structures did form as expected.","The results from experiments with two of the six markers are shown in FIGS.","3A and B and FIGS.","4A and B.","These figures show the results obtained with the second sequencing reaction protocol.","FIGS.","3A and 4A show PCR primer positions and orientations, and the expected sequences.","Note that in FIG.","4, the inner PCR primer is designed in the same orientation as forward, so the sequence reading from a sequencing reaction would be reverse complementary to the sequences highlighted with italics.","FIGS.","3B and 4B show sequencing reads from the ABI 377 sequencer analysis software.","For the 6 markers tested, we obtained sequences for five, and all of these sequences matched the expected sequences exactly.","Full length sequence reads (from the ends of inner PCR products to the ends of outer PCR products) were obtained when the standard cyclic sequencing conditions were used (data not shown).","Much shorter sequence readings (30-50bases) were obtained when the extension temperature was changed to 80° C. (as in FIGS.","3 and 4).","For 4 of the 5 makers that produced sequences the sequence started right after the inner primers, one started 3 bases away from the end of the primer.","For both sequencing conditions, the starting positions of sequencing were the same.","Example 2 Creating Sequencible Structures with Products from Extendase-Free Pfu DNA Polymerases that can be Used for SBE Genotyping Reactions The variations of starting position of the obtained sequences (Example 1) suggested that the positioning of inner PCR primers alone may not be sufficient to obtain sequencible structures with precise extending ends.","An alternative is the use of extendase-free DNA polymerases that do not add the 3′ A base at the ends of PCR products.","Thus, experiments were carried out using Pfu DNA polymerases which have been reported for lacking the extendase activity (10, 28).","Inner PCR primers were designed so that their products would be positioned right before the polymorphic site if the products formed sequencible structures.","Inner and outer PCR reactions were performed separately using Pfu DNA polymerases and the products wqere pooled together to form the sequencible structures.","Then a SBE reaction was performed using SNaPshot kit from Applied BioSystems.","SBE reactions were purified and analyzed by ABI 377 DNA sequencer using GeneScan.","Three samples were shown in FIG.","5, representing three genotypes for the marker: Homozygous A/A (upper panel), heterozygous A/G (middle panel) and homozygous G/G (lower panel).","The experiments demonstrated that Pfu DNA polymerases produce inner PCR products with precise ends that can be used to form sequencible structures for SBE reactions and to score genotypes correctly.","In a parallel experiment using Taq DNA polymerases multiple products were observed and genotypes were not clear (data not shown).","The results clearly show the differences between the two DNA polymerases.","Example 3 Control of the Reading Length for Multiple Target Sequencing The reading length of sequencing reactions is principally determined by the ratio of dNTPs/ddNTPs.","For typical sequencing applications, long sequence reads are obtained by using a high dNTPs/ddNTPs ratio (100:1) provided in commercial sequencing kits.","In the practice of the present invention, a short stretch of DNA surrounding the SNP targets will be sequenced.","For this purpose it is desirable to control the length of sequence reads so that multiple sequencing reactions can be loaded in one lane/capillary without overlapping each other.","Currently there is no commercially available kit for this purpose, but two products, the SNaPShot from Applied Biosystems and the SNuPe from Amershan Biosciences, have the potential to be modified for this purpose.","Both these kits were designed for SBE reactions, containing only dye-labeled ddNTPs.","The concentration of nucleotides in the mix is proprietary.","Individually available dye-labeled nucleotides were used to assemble a testing kit to demonstrate the principle that reading length can be controlled by dNTPs/ddNTPs ratio.","FIG.","6 shows an example of the sequencing reaction.","In this reaction, dye-labeled acyclo-ddNTPs were used to assemble the mix.","The sequencing reaction was performed using pGEM plasmid and M13-21 primer.","The ratio of dNTPs/ddNTPs and thermal cycling conditions are detailed in the figure legend.","The results showed a clean read of 30-40 bases after the primer.","This example demonstrated that the reading length can be effectively controlled by the ratio of dNTPs/dDNTPs.","To practice the current invention, especially the MulTarSeq, a specially formulated sequencing mix is therefore desirable.","Example 4 Comparison of Two-Step PCR Amplification Using Primary and Secondary Primer Sets Four SNP markers randomly were chosen from our SNP collection and PCR and nPCR primers were designed.","Both outer and inner primers had two designs, a simple design and the two domain design.","The sequences used for the common domains of the two domain were the M13 universal forward primer: cccagtcacgacgttgtaaaacg (for forward outer PCR primers) and M13 universal reverse primer: agcggataacaatttcacacagg (for reverse outer PCR primers).","For the inner PCR primer the common domain sequence was tcaGCAGCatGTCTCttcca, which contains recognition sites for two restriction enzymes (Bst71 I and Alw26 I, in capital letters).","Multiplex PCR and multiplex nPCR reactions were performed to compare the primer designs and protocols.","PCR products were then analyzed by agarose gel electrophoresis.","FIG.","7 showed the results.","Lane 1 of FIG.","7 represents a 4× multiplex PCR using simple primer design.","As ca be seen, only two bands are visible (480,510 bp).","They could not be resolved by agarose gel electrophoresis.","Lane 2 represents the same 4 amplicons, but generated using the two domain design.","In this case, the two smaller products (400, 430 bp) are clearly seen.","Note the size shifted for the larger products due to extra length (46 bp) in the two domain primers.","Lane 3 represents 4× multiplex NPCR with the same amplicons but carried out by the two-step porcedure.","In addition to the 4 outer PCR products, 4 inner PCR productscan also be seen.","This experiment demonstrates that the two domain design significantly improves both multiplex PCR and multiplex NPCR, and that the two-step procedure is especially effective to improve multiplex nPCR.","Example 5 Use of Multiplex nPCR to Create an Ordered Series of Sequencible Structures This example is provided to illustrate how a series of sequencible structures are formed from two multiplexed PCRS.","A set of five SNPs are selected and NPCR primers are designed using the two domain design.","In order to run all products on a single sequencing lane/capillary, the inner PCR products are arranged to span 150 to 400 bps range, 50 bps apart from one another.","The common domain for the inner primers should contain a restriction enzyme recognition site to produce blunt ends after the amplification.","Two 5× mulitiplexing PCRs are performed separately: one uses only the outer PCR primers the other uses only the inner PCR primers.","After the amplification an appropriate restriction enzyme is used to remove the common tag sequences from the inner end of the inner PCR products.","The outer products and the inner products then are combined, purified, denature and reannealed together.","A single base extension reaction is performed using the SNaPshot kit from Applied Biosystems.","The products then are purified and run on DNA sequencer.","The analysis of the extension products will identify the genotypes of the sample.","While the invention has been described in terms of its preferred embodiments, those skilled in the art will recognize that the invention can be practiced with modification within the spirit and scope of the appended claims.","Accordingly, the present invention 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identification.","Hum.","Mutat.","17:305-16"]],"string":"[\n [\n \"Multiple sequencible and ligatible structures for genomic analysis\",\n \"High throughput methods and kits for single nucleotide polymorphism (SNP) genotyping are provided.\",\n \"The methods involve utilizing nested PCR amplification reactions which produce sequencible and ligatible structures.\",\n \"An outer PCR primer set amplifies the SNP, and an inner PCR primer set amplifies a portion of the DNA amplified by the outer primer set, but does not amplify the SNP itself.\",\n \"The inner and outer primers may reaction include non-target common domain sequences, and the inner primer common domain sequences may comprise digestion restriction endonuclease recognition sites.\",\n \"The design of the inner primer set allows precise tailoring of the sequencible and ligatible structures with respect to length and base composition.\"\n ],\n [\n \"1.A method of producing hybrid DNA with a single strand overhang that includes a target site, comprising the steps of: obtaining a first primer which hybridizes to a 5′ strand of a strand of deoxyribonucleic acid (DNA), a second primer which hybridizes to a 3′ strand of said strand of DNA, and a third primer which hydridizes to said 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using said first primer, said second primer, and said third primer, an outer amplicon which includes said target site and an inner amplicon which excludes said target site; and forming at least one of a ligatable structure which includes a 3′-5′ sequence which excludes said target site hybridized to a 5′-3′ sequence which includes said target site, and a sequencible structure which includes a 5′-3′ sequence which excludes said target site hybridized to a 3′-5′ sequence which includes said target site.\",\n \"2.The method of claim 1 wherein said forming step forms both said ligatible structure and said sequencible structure.\",\n \"3.The method of claim 1 wherein said target site includes at least one single nucleotide polymorphism.\",\n \"4.A method of genotyping DNA of an individual by analyzing at least one target site in said DNA, comprising the steps of: obtaining a first primer which hybridizes to a 5′ strand of a strand of said DNA, a second primer which hybridizes to a 3′ strand of said strand of DNA, and a third primer which hydridizes to said 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using said first primer, said second primer, and said third primer, an outer amplicon which includes said target site and an inner amplicon which excludes said target site; and forming at least one of a sequencible structure which includes a 5′-3′ sequence which excludes said target site hybridized to a 3′-5′ sequence which includes said target site; and a ligatable structure which includes a 3′-5′ sequence which excludes said target site hybridized to a 5′-3′ sequence which includes said target site; and analyzing at least one of a sequencing product formed by sequencing said sequencible structure and a ligation product formed by ligating said ligatible structure with a labeled oligonucleotide with a DNA sequencer to determine the genotype of said individual.\",\n \"5.The method of claim 4 wherein said forming step forms both said ligatible structure and said sequencible structure.\",\n \"6.The method of claim 4 wherein said at least one target site includes a single nucleotide polymorphism.\",\n \"7.The method of claim 4 wherein said sequencible structure is sequenced by a technique selected from the group consisting of dideoxy sequencing, pyrosequencing, and single base extension.\",\n \"8.The method of claim 4 wherein a plurality of target sites are simultaneously analyzed by the step of producing an ordered series of sequencing products of varying, non-overlapping length each specific for a particular target site, and wherein said step of analyzing is carried out by electrophoresing said ordered series of sequencing products in a single channel of said DNA sequencer.\",\n \"9.The method of claim 8 wherein said step of producing an ordered series of sequencing products is carried out by single base extension (SBE).\",\n \"10.The method of claim 8 wherein said step of producing an ordered series of sequencing products is carried out by a dideoxy sequencing reaction utilizing a low ratio of dNTPs to ddNTPs.\",\n \"11.The method of claim 4 wherein said labeled oligonucleotide is fluorescently labeled.\",\n \"12.The method of claim 4 wherein a plurality of ligatable structures are formed in said forming step, each being specific for one of a plurality of target sites, and said ligation products are of varying, non-overlapping lengths with said labeled oligonucleotides being degenerate, said step of analyzing is carried out by electrophoresing a plurality of ligation products in a single channel of said DNA sequencer.\",\n \"13.A method for analyzing at least one target site in a DNA molecule, comprising the steps of amplifying by nested PCR said target site, wherein said nested PCR is carried out using inner and outer PCR primer pairs, wherein said outer PCR primer pair forms a first PCR product which contains said target site, and wherein said inner PCR primer pair forms a second PCR product which contains a portion of said first PCR product but does not contain said target site, denaturing said first and said second PCR products to form ssDNA sequences, reannealing said ss DNA sequences to form a sequencible hybrid DNA molecule and a ligatible hybrid DNA molecule, performing sequencing reactions with said sequencible hybrid DNA molecule and ligation reactions with said ligatible hybrid DNA molecule, and determining the characteristics of said target site by analyzing results obtained in said performing step.\",\n \"14.The method of claim 13 wherein said at least one target site is an SNP polymorphism site.\",\n \"15.The method of claim 13 wherein said inner and outer primer pairs comprise a sequence tag.\",\n \"16.The method of claim 15 wherein said sequence tag comprises a restriction enzyme recognition site.\",\n \"17.The method of claim 13 wherein said step of amplifying is carried out using a low concentration of primers, and further comprising a second step of amplifying, wherein said second step of amplifying uses secondary primers for amplification of said sequence tags.\",\n \"18.The method of claim 13 wherein one target site is analyzed.\",\n \"19.The method of claim 13 wherein a plurality of target sites are analyzed.\",\n \"20.The method of claim 13 wherein said step of amplifying is carried out in a single multiplex PCR reaction.\",\n \"21.The method of claim 13 wherein said step of amplifying is carried out in multiple independent PCR reactions.\",\n \"22.The method of claim 13 wherein said results obtained in said performing step are analyzed by a DNA sequencer.\",\n \"23.Inner and outer PCR primer pairs for the amplification of a target site in a DNA molecule, wherein said outer PCR primer pair forms a first PCR product which contains said target site, and wherein said inner PCR primer pair forms a second PCR product which contains a portion of said first PCR product but does not contain said target site.\",\n \"24.The primer pairs of claim 23, wherein said target site is an SNP polymorphism site.\",\n \"25.The primer pairs of claim 23, wherein said primer pairs comprise a sequence tag.\",\n \"26.A kit for amplification of at least one target site in a DNA molecule, comprising inner and outer PCR primer pairs for the amplification of said target site, wherein said outer primer pair amplifies a portion of said DNA molecule including said target site, and wherein said inner primer pair amplifies part of said portion of said DNA molecule amplified by said outer primer pair and excludes said target site.\",\n \"27.The kit of claim 26 wherein said inner and outer PCR primer pairs comprise sequence tags.\",\n \"28.The kit of claim 27, further comprising secondary primers for the amplification of said sequence tags.\",\n \"29.A dideoxy DNA sequencing kit for producing short chain termination fragments, comprising dNTPs and ddNTPs present in a dNTP:ddNTP ratio ranging from 1:0 to 1:10.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>1.Field of the Invention The invention generally relates to single nucleotide polymorphism (SNP) genotyping.\",\n \"In particular, the invention provides a method for SNP genotyping based on a nested PCR design that creates structures directly suitable for both DNA sequencing and ligation reactions.\",\n \"2.Background of the Invention With the completion of the rough draft of the human genome, over 1.5 million non-redundant SNPs have been identified and mapped by the publicly funded project and the SNP consortium (Sachidanandam et al, 2001).\",\n \"The availability of such a large number of SNPs has prompted great interest in SNP related applications and research, such as large scale linkage, genomic association and pharmarcogenetic studies.\",\n \"SNPs entered the central arena of genetics due to their abundance (Collins et al, 1997; Sachidanandam et al, 2001), stability (Sachidanandam et al, 2001) and the relative ease (Kwok 2000; Shi 2001; Syvanen 2002) with which they are genotyped.\",\n \"The targets of SNP genotyping are normally small pieces of DNA, ranging from 150-300 base pairs.\",\n \"Based on the SNP consortium report, the average SNP density in the human genome appears to be greater than 1 SNP/KB.\",\n \"This high density provides a tool to analyze genomic structure at very high resolution, and to establish a direct correlation between genetic coding variation and biological function.\",\n \"The relative stability of SNPs (compared to mini- and micro-satellite markers) in evolution makes it simpler to carry out this task.\",\n \"Although SNP genotyping is generally easier than microsatellite genotyping, and despite the rapid and significant advances in SNP genotyping technology in the last few years, major issues remain unsolved.\",\n \"Whereas only a few years ago, the main concern was the need to have a large number of SNPs available, currently the most urgent issues are increasing throughput and decreasing cost.\",\n \"For example, for large scale applications as projected by Kruglyak (1999) and Long (1999), a reasonable cost would be <$0.10/genotype, or else large-scale projects will become prohibitively expensive.\",\n \"To fulfill the goals of understanding the genetics of complex traits and common diseases, cost-effective and higher throughput methodology in SNP genotyping is essential.\",\n \"Typically, genotyping protocols have three components: target amplification, allelic discrimination and product detection and identification.\",\n \"They are normally executed sequentially, but can be processed in a single step reaction, depending on the means which are used for allele discrimination and signal detection.\",\n \"Such single step procedures are well-suited to automation, but are not necessarily effective in terms of cost and throughput.\",\n \"PCR is the dominant procedure utilized for target amplification.\",\n \"As is widely known, a PCR can readily amplify targets present in lower copy number by a factor of 108 or more.\",\n \"With respect to allele discrimination, all SNP genotyping technologies currently available are based on mechanisms involving one or more of the following: DNA polymerase, hybridization and DNA ligase.\",\n \"DNA polymerase methods include single base extension (SBE) (Armstrong, 2000; Barta 2001; Bray 2001; Cai 2001; Chen 1997; Chem 1999; Chen 1998; Chen 1997; Chen 1997; Fan 2000; Lindblad-Toh 2000; Nikiforov 1994; Pastinen 1996; Pastinen 2000; Ross, 1998; Sauer 2000; Syvanen 1999; Ye 2001), de novo sequencing including pyrosequencing (Nordstrom 2000; Ronaghi 2001), allele-specific PCR and extension (Germer 1999; Myakishev 2001; Pastinen 2000), and structure-specific cleavage (Fors 2000).\",\n \"Of these different forms, SBE is the most widely used and has been adapted for many different detection platforms.\",\n \"Hybridization is also widely used in several different forms, including dynamic hybridization (Prince 2001), and is the primary method currently used in all microarray detection formats.\",\n \"DNA ligase methods are based on the ability of DNA ligase to join the ends of two oligonucleotides annealed next to each other on a template (Tong 2000; Tong1999).\",\n \"Two oligonucleotides can be designed to anneal to both sides of a SNP site, and by detecting the formation of ligation product, the genotype of a target can be inferred (Chen 1998; Delahunty 1996; Gerry 1999; Iannone 2000).\",\n \"Once a target DNA sequence has been amplified and the allelic variants discriminated, the next step in a generic genotyping protocol is to detect and identify the allele specific products.\",\n \"Detection mechanisms vary greatly, from simple fluorescence intensity (Armstrong 2000; Cai 2000; Chen 1998; Chen 1997; Delahunty 1996; Dubertret 2001; Fan 2000; Fergusoon 2000; Fors 2000; Germer 1999; Germer 2000; Gerry 1999; Iannone 2000; Lindblad-Toh 2000; Lindroos 2001; Marras 1999; Medintz 2001; Myakishev 2001; Nikiforov 1994; Pastinen 1997; Pastinen 2000; Prince 2001; Syvanen 1999; Ye 2001) to very precise mass (Bray 2001; Ross 1998; Sauer 2000) or electric charge (Gilles 1999; Woolley 2000) measurement.\",\n \"The detection mechanisms roughly fall into two categories: homogeneous and solid phase mediated detection.\",\n \"All homogeneous detection platforms depend on measuring fluorescence intensities and their change during and/or after reactions.\",\n \"One common feature among homogeneous approaches is that they do not require any separation/purification prior to signal acquisition, making them amenable to automation.\",\n \"Solid support techniques include flow cytometry genotyping (Armstrong 2000; Cai 2000; Ye 2001), zip-code microarrays (Fan 2000; Gerry 1999), and mass spectrometry genotyping (Bray 2001; Ross 1998; Sauer 2000).\",\n \"Using a solid support in the detection step may potentially increase throughput and reduce cost, but unfortunately also entails the risk of complicating protocols and compromising data quality.\",\n \"For example, when reaction mixtures are applied to a solid support, unintended binding of fluorescence dye to the solid surface can occur, necessitating extensive washing to minimize spurious signals.\",\n \"With respect to cost, prices for genotyping technologies currently available on the market vary considerably, from $0.50-2.00/genotype.\",\n \"For example, the template-directed dye-terminator incorporation assay with fluorescence detection (FP-TDI) (Chen 1999) costs roughly $0.50/genotype for reagents by list price.\",\n \"For other methods, due to their requirement for special reagents and/or clean-up procedures, the cost is higher, e.g.\",\n \"MALDI-TOF mass spectrometry is about $0.75-1.00, whereas pyrosequencing approaches $2.00/genotype.\",\n \"No currently available technology approaches the low cost which is necessary in order to prevent large-scale projects from becoming prohibitively expensive, i.e.\",\n \"<$0.10/genotype.\",\n \"There are currently many genotyping applications in use or under development that require a relatively large number of both SNPs and samples (e.g., both in the thousands or more), demanding both cost effective and high throughput technologies.\",\n \"Examples include gene mapping studies by linkage or linkage disequilibrium (Kruglyak 1999; Long 1999) for complex traits and pharmacogenetics (Riley 2000).\",\n \"Clearly, these applications are critical to improving our understanding of the genetics of complex traits, common diseases and drug response, and to help attain the goal of individualized medicine.\",\n \"However, none of the current approaches are suitable for such applications in terms of cost effectiveness coupled with high throughput potential, and SNP genotyping demands both.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The present invention provides a methodology for high-throughput SNP genotyping that is highly cost effective.\",\n \"This novel approach makes use of the high throughput capacity of DNA sequencers for SNP genotyping and is based on a nested PCR design that creates a series of ordered “structures” in parallel and exploits them for SNP genotyping assays.\",\n \"The invention thus provides novel genotyping technologies that significantly increase throughput, reduce cost and are accessible for most researchers, leading to significant improvements in genotyping technology.\",\n \"While typical SNP genotyping approaches currently available score <10 SNPs simultaneously, the methods of the present invention increase the number to about 50-100 with obvious advantages for throughput and cost.\",\n \"The present invention provides a method of producing hybrid DNA with a single strand overhang that includes a target sequence.\",\n \"The steps of the method include: obtaining a first primer which hybridizes to a 5′ strand of a strand of deoxyribonucleic acid (DNA), a second primer which hybridizes to a 3′ strand of the strand of DNA, and a third primer which hydridizes to said 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using the first primer, the second primer, and the third primer, an outer amplicon which includes a target sequence and an inner amplicon which excludes the target sequence; forming at least one of the following: a ligatable structure which includes a 3′-5′ sequence which excludes the target sequence hybridized to a 5′-3′ sequence which includes the target sequence, and a sequencible structure which includes a 5′-3′ sequence which excludes the target sequence hybridized to a 3′-5′ sequence which includes the target sequence; sequencing the sequencible structure(s) and ligating the ligatible structure(s) with a labeled oligonucleotide; and analyzing the sequencing products and ligation products with a DNA sequencer to determine the genotype of said individual.\",\n \"The method may form both the ligatible structure and the sequencible structure.\",\n \"The target sequence(s) may include a single nucleotide polymorphism.\",\n \"The invention also provides a method of genotyping the deoxyribonucleic acid (DNA) of an individual by analyzing at least on target sequence in the DNA.\",\n \"The steps of the method include: obtaining a first primer which hybridizes to a 5′ strand of a strand of said DNA, a second primer which hybridizes to a 3′ strand of the strand of DNA, and a third primer which hydridizes to the 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using the first primer, the second primer, and the third primer, an outer amplicon which includes said target sequence and an inner amplicon which excludes the target sequence; and forming at least one of the following: a ligatable structure which includes a 3′-5′ sequence which excludes the target sequence hybridized to a 5′-3′ sequence which includes the target sequence, and a sequencible structure which includes a 5′-3′ sequence which excludes the target sequence hybridized to a 3′-5′ sequence which includes the target sequence; sequencing the sequencible structure and ligating the ligatible structure with a labeled oligonucleotide; and analyzing the sequencing and ligation products so produced with a DNA sequencer to determine the genotype of the individual.\",\n \"The forming step of the method may form both the ligatible structure and the sequencible structure.\",\n \"The target sequence(s) may include a single nucleotide polymorphism.\",\n \"The sequencible structure may be sequenced by a technique such as dideoxy sequencing, pyrosequencing, and single base extension.\",\n \"In a preferred embodiment, of the method, i) a plurality of target sequences is analyzed; ii) the step of forming forms an ordered series of sequencible structures; iii) the step of sequencing produces an ordered series of sequencing products of varying, non-overlapping length; and iv) the step of analyzing is carried out by electrophoresing the ordered series of sequencing products in a single channel of said DNA sequencer.\",\n \"In some embodiments, the step of sequencing is carried out by single base extension (SBE).\",\n \"In other embodiments, the step of sequencing is carried out by a dideoxy sequencing reaction utilizing a ratio of dNTPs to ddNTPs that is lower than that which is typically used in order to produce a short sequence reading.\",\n \"In a preferred embodiment of the present invention, the labeled oligonucleotide is fluorescently labeled.\",\n \"In one embodiment of the method, i) a plurality of target sequences is analyzed; ii) the labeled oligonucleotides are degenerate; iii) the ligation products are of varying, non-overlapping lengths; and iv) the step of analyzing is carried out by electrophoresing a plurality of ligation products in a single channel of said DNA sequencer.\",\n \"The present invention also provides a method for analyzing at least one target site in a DNA molecule.\",\n \"The method includes the steps of: 1) amplifying the target site(s) by nested PCR; (The nested PCR is carried out using inner and outer PCR primer pairs, wherein the outer PCR primer pair forms a first PCR product which contains a target site, and the inner PCR primer pair forms a second PCR product which contains a portion of the first PCR product but does not contain the target site); 2) denaturing the first and said second PCR products to form ssDNA sequences, 3) reannealing the ss DNA sequences to form a sequencible hybrid DNA molecule and a ligatible hybrid DNA molecule, 4) performing sequencing reactions with said sequencible hybrid DNA molecule and ligation reactions with said ligatible hybrid DNA molecule, and 5) determining the characteristics of the target sequence by analyzing results obtained in the performing step.\",\n \"In a preferred embodiment of the method, the target site(s) may be SNP polymorphism sites.\",\n \"Further, the inner and outer primer pairs may comprise a sequence tag, which in turn may comprises a restriction enzyme recognition site.\",\n \"In some embodiments of the method, the step of amplifying may be carried out using a low concentration of primers, and may further comprising a second step of amplification using secondary primers for amplification of the sequence tags.\",\n \"According to the method, one or several target sites may be analyzed.\",\n \"The step of amplifying may be carried out in a single multiplex PCR reaction, or in multiple independent PCR reactions, and the results may be analyzed by a DNA sequencer.\",\n \"The invention also provides inner and outer PCR primer pairs for the amplification of a target sequence in a DNA molecule, wherein the outer PCR primer pair forms a first PCR product which contains the target sequence, and the inner PCR primer pair forms a second PCR product which contains a portion of the first PCR product but does not contain said target sequence.\",\n \"The target sequence may be an SNP polymorphism site.\",\n \"The primer pairs may comprise a sequence tag.\",\n \"The present invention also provides a kit for amplification of at least one target sequence in a DNA molecule.\",\n \"The kit includes inner and outer PCR primer pairs for the amplification of the target sequence.\",\n \"The outer primer pair amplifies a portion of the DNA molecule which includes the target sequence, and the inner primer pair amplifies part of the same portion of the DNA molecule that is amplified by the outer primer pair, but excludes the target sequence.\",\n \"The inner and outer PCR primer pairs may comprise sequence tags.\",\n \"The kit may further comprise secondary primers for the amplification of the sequence tags.\",\n \"The present invention also provides a dideoxy DNA sequencing kit for producing short chain termination fragments.\",\n \"The kit includes dNTPs and ddNTPs which are present in a low dNTP:ddNTP ratio.\"\n ],\n [\n \"BACKGROUND OF THE INVENTION 1.Field of the Invention The invention generally relates to single nucleotide polymorphism (SNP) genotyping.\",\n \"In particular, the invention provides a method for SNP genotyping based on a nested PCR design that creates structures directly suitable for both DNA sequencing and ligation reactions.\",\n \"2.Background of the Invention With the completion of the rough draft of the human genome, over 1.5 million non-redundant SNPs have been identified and mapped by the publicly funded project and the SNP consortium (Sachidanandam et al, 2001).\",\n \"The availability of such a large number of SNPs has prompted great interest in SNP related applications and research, such as large scale linkage, genomic association and pharmarcogenetic studies.\",\n \"SNPs entered the central arena of genetics due to their abundance (Collins et al, 1997; Sachidanandam et al, 2001), stability (Sachidanandam et al, 2001) and the relative ease (Kwok 2000; Shi 2001; Syvanen 2002) with which they are genotyped.\",\n \"The targets of SNP genotyping are normally small pieces of DNA, ranging from 150-300 base pairs.\",\n \"Based on the SNP consortium report, the average SNP density in the human genome appears to be greater than 1 SNP/KB.\",\n \"This high density provides a tool to analyze genomic structure at very high resolution, and to establish a direct correlation between genetic coding variation and biological function.\",\n \"The relative stability of SNPs (compared to mini- and micro-satellite markers) in evolution makes it simpler to carry out this task.\",\n \"Although SNP genotyping is generally easier than microsatellite genotyping, and despite the rapid and significant advances in SNP genotyping technology in the last few years, major issues remain unsolved.\",\n \"Whereas only a few years ago, the main concern was the need to have a large number of SNPs available, currently the most urgent issues are increasing throughput and decreasing cost.\",\n \"For example, for large scale applications as projected by Kruglyak (1999) and Long (1999), a reasonable cost would be <$0.10/genotype, or else large-scale projects will become prohibitively expensive.\",\n \"To fulfill the goals of understanding the genetics of complex traits and common diseases, cost-effective and higher throughput methodology in SNP genotyping is essential.\",\n \"Typically, genotyping protocols have three components: target amplification, allelic discrimination and product detection and identification.\",\n \"They are normally executed sequentially, but can be processed in a single step reaction, depending on the means which are used for allele discrimination and signal detection.\",\n \"Such single step procedures are well-suited to automation, but are not necessarily effective in terms of cost and throughput.\",\n \"PCR is the dominant procedure utilized for target amplification.\",\n \"As is widely known, a PCR can readily amplify targets present in lower copy number by a factor of 108 or more.\",\n \"With respect to allele discrimination, all SNP genotyping technologies currently available are based on mechanisms involving one or more of the following: DNA polymerase, hybridization and DNA ligase.\",\n \"DNA polymerase methods include single base extension (SBE) (Armstrong, 2000; Barta 2001; Bray 2001; Cai 2001; Chen 1997; Chem 1999; Chen 1998; Chen 1997; Chen 1997; Fan 2000; Lindblad-Toh 2000; Nikiforov 1994; Pastinen 1996; Pastinen 2000; Ross, 1998; Sauer 2000; Syvanen 1999; Ye 2001), de novo sequencing including pyrosequencing (Nordstrom 2000; Ronaghi 2001), allele-specific PCR and extension (Germer 1999; Myakishev 2001; Pastinen 2000), and structure-specific cleavage (Fors 2000).\",\n \"Of these different forms, SBE is the most widely used and has been adapted for many different detection platforms.\",\n \"Hybridization is also widely used in several different forms, including dynamic hybridization (Prince 2001), and is the primary method currently used in all microarray detection formats.\",\n \"DNA ligase methods are based on the ability of DNA ligase to join the ends of two oligonucleotides annealed next to each other on a template (Tong 2000; Tong1999).\",\n \"Two oligonucleotides can be designed to anneal to both sides of a SNP site, and by detecting the formation of ligation product, the genotype of a target can be inferred (Chen 1998; Delahunty 1996; Gerry 1999; Iannone 2000).\",\n \"Once a target DNA sequence has been amplified and the allelic variants discriminated, the next step in a generic genotyping protocol is to detect and identify the allele specific products.\",\n \"Detection mechanisms vary greatly, from simple fluorescence intensity (Armstrong 2000; Cai 2000; Chen 1998; Chen 1997; Delahunty 1996; Dubertret 2001; Fan 2000; Fergusoon 2000; Fors 2000; Germer 1999; Germer 2000; Gerry 1999; Iannone 2000; Lindblad-Toh 2000; Lindroos 2001; Marras 1999; Medintz 2001; Myakishev 2001; Nikiforov 1994; Pastinen 1997; Pastinen 2000; Prince 2001; Syvanen 1999; Ye 2001) to very precise mass (Bray 2001; Ross 1998; Sauer 2000) or electric charge (Gilles 1999; Woolley 2000) measurement.\",\n \"The detection mechanisms roughly fall into two categories: homogeneous and solid phase mediated detection.\",\n \"All homogeneous detection platforms depend on measuring fluorescence intensities and their change during and/or after reactions.\",\n \"One common feature among homogeneous approaches is that they do not require any separation/purification prior to signal acquisition, making them amenable to automation.\",\n \"Solid support techniques include flow cytometry genotyping (Armstrong 2000; Cai 2000; Ye 2001), zip-code microarrays (Fan 2000; Gerry 1999), and mass spectrometry genotyping (Bray 2001; Ross 1998; Sauer 2000).\",\n \"Using a solid support in the detection step may potentially increase throughput and reduce cost, but unfortunately also entails the risk of complicating protocols and compromising data quality.\",\n \"For example, when reaction mixtures are applied to a solid support, unintended binding of fluorescence dye to the solid surface can occur, necessitating extensive washing to minimize spurious signals.\",\n \"With respect to cost, prices for genotyping technologies currently available on the market vary considerably, from $0.50-2.00/genotype.\",\n \"For example, the template-directed dye-terminator incorporation assay with fluorescence detection (FP-TDI) (Chen 1999) costs roughly $0.50/genotype for reagents by list price.\",\n \"For other methods, due to their requirement for special reagents and/or clean-up procedures, the cost is higher, e.g.\",\n \"MALDI-TOF mass spectrometry is about $0.75-1.00, whereas pyrosequencing approaches $2.00/genotype.\",\n \"No currently available technology approaches the low cost which is necessary in order to prevent large-scale projects from becoming prohibitively expensive, i.e.\",\n \"<$0.10/genotype.\",\n \"There are currently many genotyping applications in use or under development that require a relatively large number of both SNPs and samples (e.g., both in the thousands or more), demanding both cost effective and high throughput technologies.\",\n \"Examples include gene mapping studies by linkage or linkage disequilibrium (Kruglyak 1999; Long 1999) for complex traits and pharmacogenetics (Riley 2000).\",\n \"Clearly, these applications are critical to improving our understanding of the genetics of complex traits, common diseases and drug response, and to help attain the goal of individualized medicine.\",\n \"However, none of the current approaches are suitable for such applications in terms of cost effectiveness coupled with high throughput potential, and SNP genotyping demands both.\",\n \"SUMMARY OF THE INVENTION The present invention provides a methodology for high-throughput SNP genotyping that is highly cost effective.\",\n \"This novel approach makes use of the high throughput capacity of DNA sequencers for SNP genotyping and is based on a nested PCR design that creates a series of ordered “structures” in parallel and exploits them for SNP genotyping assays.\",\n \"The invention thus provides novel genotyping technologies that significantly increase throughput, reduce cost and are accessible for most researchers, leading to significant improvements in genotyping technology.\",\n \"While typical SNP genotyping approaches currently available score <10 SNPs simultaneously, the methods of the present invention increase the number to about 50-100 with obvious advantages for throughput and cost.\",\n \"The present invention provides a method of producing hybrid DNA with a single strand overhang that includes a target sequence.\",\n \"The steps of the method include: obtaining a first primer which hybridizes to a 5′ strand of a strand of deoxyribonucleic acid (DNA), a second primer which hybridizes to a 3′ strand of the strand of DNA, and a third primer which hydridizes to said 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using the first primer, the second primer, and the third primer, an outer amplicon which includes a target sequence and an inner amplicon which excludes the target sequence; forming at least one of the following: a ligatable structure which includes a 3′-5′ sequence which excludes the target sequence hybridized to a 5′-3′ sequence which includes the target sequence, and a sequencible structure which includes a 5′-3′ sequence which excludes the target sequence hybridized to a 3′-5′ sequence which includes the target sequence; sequencing the sequencible structure(s) and ligating the ligatible structure(s) with a labeled oligonucleotide; and analyzing the sequencing products and ligation products with a DNA sequencer to determine the genotype of said individual.\",\n \"The method may form both the ligatible structure and the sequencible structure.\",\n \"The target sequence(s) may include a single nucleotide polymorphism.\",\n \"The invention also provides a method of genotyping the deoxyribonucleic acid (DNA) of an individual by analyzing at least on target sequence in the DNA.\",\n \"The steps of the method include: obtaining a first primer which hybridizes to a 5′ strand of a strand of said DNA, a second primer which hybridizes to a 3′ strand of the strand of DNA, and a third primer which hydridizes to the 3′ strand of DNA; producing by nested polymerase chain reaction (PCR) using the first primer, the second primer, and the third primer, an outer amplicon which includes said target sequence and an inner amplicon which excludes the target sequence; and forming at least one of the following: a ligatable structure which includes a 3′-5′ sequence which excludes the target sequence hybridized to a 5′-3′ sequence which includes the target sequence, and a sequencible structure which includes a 5′-3′ sequence which excludes the target sequence hybridized to a 3′-5′ sequence which includes the target sequence; sequencing the sequencible structure and ligating the ligatible structure with a labeled oligonucleotide; and analyzing the sequencing and ligation products so produced with a DNA sequencer to determine the genotype of the individual.\",\n \"The forming step of the method may form both the ligatible structure and the sequencible structure.\",\n \"The target sequence(s) may include a single nucleotide polymorphism.\",\n \"The sequencible structure may be sequenced by a technique such as dideoxy sequencing, pyrosequencing, and single base extension.\",\n \"In a preferred embodiment, of the method, i) a plurality of target sequences is analyzed; ii) the step of forming forms an ordered series of sequencible structures; iii) the step of sequencing produces an ordered series of sequencing products of varying, non-overlapping length; and iv) the step of analyzing is carried out by electrophoresing the ordered series of sequencing products in a single channel of said DNA sequencer.\",\n \"In some embodiments, the step of sequencing is carried out by single base extension (SBE).\",\n \"In other embodiments, the step of sequencing is carried out by a dideoxy sequencing reaction utilizing a ratio of dNTPs to ddNTPs that is lower than that which is typically used in order to produce a short sequence reading.\",\n \"In a preferred embodiment of the present invention, the labeled oligonucleotide is fluorescently labeled.\",\n \"In one embodiment of the method, i) a plurality of target sequences is analyzed; ii) the labeled oligonucleotides are degenerate; iii) the ligation products are of varying, non-overlapping lengths; and iv) the step of analyzing is carried out by electrophoresing a plurality of ligation products in a single channel of said DNA sequencer.\",\n \"The present invention also provides a method for analyzing at least one target site in a DNA molecule.\",\n \"The method includes the steps of: 1) amplifying the target site(s) by nested PCR; (The nested PCR is carried out using inner and outer PCR primer pairs, wherein the outer PCR primer pair forms a first PCR product which contains a target site, and the inner PCR primer pair forms a second PCR product which contains a portion of the first PCR product but does not contain the target site); 2) denaturing the first and said second PCR products to form ssDNA sequences, 3) reannealing the ss DNA sequences to form a sequencible hybrid DNA molecule and a ligatible hybrid DNA molecule, 4) performing sequencing reactions with said sequencible hybrid DNA molecule and ligation reactions with said ligatible hybrid DNA molecule, and 5) determining the characteristics of the target sequence by analyzing results obtained in the performing step.\",\n \"In a preferred embodiment of the method, the target site(s) may be SNP polymorphism sites.\",\n \"Further, the inner and outer primer pairs may comprise a sequence tag, which in turn may comprises a restriction enzyme recognition site.\",\n \"In some embodiments of the method, the step of amplifying may be carried out using a low concentration of primers, and may further comprising a second step of amplification using secondary primers for amplification of the sequence tags.\",\n \"According to the method, one or several target sites may be analyzed.\",\n \"The step of amplifying may be carried out in a single multiplex PCR reaction, or in multiple independent PCR reactions, and the results may be analyzed by a DNA sequencer.\",\n \"The invention also provides inner and outer PCR primer pairs for the amplification of a target sequence in a DNA molecule, wherein the outer PCR primer pair forms a first PCR product which contains the target sequence, and the inner PCR primer pair forms a second PCR product which contains a portion of the first PCR product but does not contain said target sequence.\",\n \"The target sequence may be an SNP polymorphism site.\",\n \"The primer pairs may comprise a sequence tag.\",\n \"The present invention also provides a kit for amplification of at least one target sequence in a DNA molecule.\",\n \"The kit includes inner and outer PCR primer pairs for the amplification of the target sequence.\",\n \"The outer primer pair amplifies a portion of the DNA molecule which includes the target sequence, and the inner primer pair amplifies part of the same portion of the DNA molecule that is amplified by the outer primer pair, but excludes the target sequence.\",\n \"The inner and outer PCR primer pairs may comprise sequence tags.\",\n \"The kit may further comprise secondary primers for the amplification of the sequence tags.\",\n \"The present invention also provides a dideoxy DNA sequencing kit for producing short chain termination fragments.\",\n \"The kit includes dNTPs and ddNTPs which are present in a low dNTP:ddNTP ratio.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1.A schematic drawing illustrates a nested PCR design that would create both a sequencible and a ligatible structure.\",\n \"The reaction uses two primer sets (inner and outer).\",\n \"P1, forward outer and forward inner primer; P2, reverse inner primer; P3, reverse outer primer.\",\n \"When this design is multiplexed, multiple sequencible and a ligatible structures are formed.\",\n \"These structures can be used for genotyping reactions.\",\n \"FIGS.\",\n \"2A and B.\",\n \"A, a multiplex PCR scheme that amplifies three target sites.\",\n \"Two primer sets (inner and outer) are used for the amplification of each target site.\",\n \"The target sites are represented by “x”.\",\n \"All primers contain two domains, a common domain and a target specific domain.\",\n \"The target specific domains, indicated by arrows, have sequences unique to the targets, whereas the common domains are not target specific.\",\n \"=common domain of forward inner and forward outer primers; ▮=common domain of inner reverse primers, - - - =common domain of reverse outer primers.\",\n \"B, close-up view of common domain of inner primer shows restriction sites incorporated into the sequence of the domain.\",\n \"FIGS.\",\n \"3A and B.\",\n \"A. Depiction of nested PCR primer design.\",\n \"The sequence (SEQ ID NO.\",\n \"1) represents 393 nucleotides from human chromosome 6.Nucleotides corresponding to primer pairs are shaded with gray, and arrows indicate primer orientation.\",\n \"Arrow indicates site of polymorphism.\",\n \"The sequence which was generated by ABI 377 sequencer is shown in italics, and is also depicted in FIG.\",\n \"3B (SEQ ID NO.\",\n \"2).\",\n \"The sample is a A/A homozygote.\",\n \"FIGS.\",\n \"4A and B.\",\n \"A. Depiction of nested PCR primer design.\",\n \"The sequence (SEQ ID NO.\",\n \"3) represents 494 nucleotides from human chromosome 6.Nucleotides corresponding to primer pairs are shaded with gray, and arrows indicate primer orientation.\",\n \"Arrow indicates site of polymorphism.\",\n \"The sequence which was generated by ABI 377 sequencer is shown in italics, and is also depicted in FIG.\",\n \"4B (SEQ ID NO.\",\n \"4).\",\n \"Two samples were shown in FIG.\",\n \"4B.\",\n \"The upper panel was a heterozygote (C/T), at base position 24, as pointed by the arrow, both bases © and T) were identified.\",\n \"The lower panel was a homozygote (T/T), at the indicated position only the T base was identified.\",\n \"The example demonstrated that a single nucleotide polymorphism (SNP) could be identified by sequencing from the sequencible structure formed from the nPCR design.\",\n \"FIG.\",\n \"5.SBE genotyping using sequencible structures from nPCR products amplified by Pfu DNA polymerases.\",\n \"Nested PCR primers were designed for an SNP marker from human chromosome 6.The inner product size is 189 bp, its 3′ end is positioned immediately upstream to the polymorphic site when it forms sequencible structures.\",\n \"PCRs were performed with DNA samples of known genotypes.\",\n \"After PCRs the outer and inner products were purified and denatured together to form sequencible structures.\",\n \"SBE reactions were performed and products analyzed in ABI 377 Sequencer using Genescan.\",\n \"Three samples were presented.\",\n \"The upper panel is a homozygous A/A where an SBE product of 190 bp is found (arrow).\",\n \"Products of the same size are found in the heterozygous (middle panel) and the other homozygous (lower panel).\",\n \"The peaks after the dye front are ROX500 size makers (35, 50, 75, 100, 139, 150, 160, 200, 250 bp respectively).\",\n \"FIG.\",\n \"6.An example illustrates a short sequence read with a specially assembled sequencing mixture that has a different ddNTPs/dNTPs ratio.\",\n \"The components used for the example were the following: 1.75 μM of −21 M13 primer, 300 ng of pGEM, 60 μM of each dNTPs, 2.5 μM of R6G-acyclo-ATP, 12 μM of ROX-acyclo-CTP, 10 μM of BODIPY-Fluorescein-acyclo-GTP, 25 μM of TAMRA-acyclo-UTP, 1 unit of AcycloPol DNA polymerase and 1× AcycloPol reaction buffer.\",\n \"The thermal cycling conditions were 96° C. for 3 min., followed by 25 cycles of 96° C. for 1 min., 50° C. for 15 sec.\",\n \"and 60° C. for 4 min.\",\n \"FIG.\",\n \"7.Primer designs affected multiplex PCR and nPCR.\",\n \"Lane 1 was a 4×multiplex PCR with simple PCR design, Lane 2 was the same 4 amplicons but with the two domain design, and Lane 3 was the 4× multiplex nPCR with the same amplicons performed in two sequential reactions.\",\n \"In the first reaction, equal amount of primary primers at 10 nM were used.\",\n \"The cycling condition was 95° C. for 10 min, followed by 10 cycles of 95° C. for 45 sec., 65° C. for 5 sec., ramping to 55° C. at 0.1° C./sec and 55° C. for 90 sec.\",\n \"For the second reaction the primers used were different among the three lanes.\",\n \"Lane 1, since it did not have secondary primers, the same primers were used as in first reaction but the concentration increased to 100 nM.\",\n \"Lane 2 used two secondary primers, M13 forward and reverse, each 400 nM.\",\n \"Lane 3 used three primers, M13 forward (600 nM), M13 reverse (400 nM) and the common tag for inner PCR primers (400 nM).\",\n \"The cycling conditions for the second reaction was following: 95° C. for 10 min, followed by 30 cycles of 95° C. for 45 sec., 55° C. for 90 sec.\",\n \"DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION The present invention provides methods of high-throughput SNP genotyping that are highly cost-effective.\",\n \"The methods utilize the high-throughput capability of ubiquitous and readily accessible DNA sequencers in order to carry out the SNP analysis.\",\n \"This is possible due to the experimental design of the methods, which are based on nested PCR reactions.\",\n \"In a preferred embodiment, the target sites which are analyzed are SNP polymorphism sites.\",\n \"However, those of skill in the art will recognize that other target sites may also be analyzed by the methods of the present invention.\",\n \"For example, short insertions and deletions, and closely spaced multiple SNPs can be identified by the multiple target sequencing approaches disclosed herein.\",\n \"As is well-known to those of skill in the art, nested PCR is a PCR amplification method that “nests” one PCR reaction within another.\",\n \"Two sets of primers are used: “outer” primers amplify a portion of a DNA strand, and “inner” primers amplify a smaller portion of the same DNA that is located within the larger, outer portion.\",\n \"Thus, two PCR products (amplicons) are produced.\",\n \"One amplicon is smaller than the other and contains a “subset” of the bases contained in the larger, outer amplicon.\",\n \"In the practice of the present invention, a new design for nested PCR is used.\",\n \"Primers are designed so that the outer amplicon includes a target site (e.g.\",\n \"an SNP polymorphism site) and the inner amplicon excludes the target site.\",\n \"The purpose of the inner PCR in the nested reaction is to synthesize a DNA fragment whose size and position relative to the outer PCR can be precisely controlled.\",\n \"This design concept is illustrated in FIG.\",\n \"1.As can be seen, in FIG.\",\n \"1, the inner primer set is composed of primers P1 and P2, and the outer primer set is composed of primers P1 and P3.\",\n \"(In this example, Primer P1 is utilized in both primer sets.)\",\n \"The P1-P3 outer primer set flanks the indicated target site whereas the P1-P2 inner primer pair does not.\",\n \"PCR amplification of the DNA with these primers will produce two amplicons.\",\n \"The larger of the two is the outer amplicon formed by amplification with the P1-P3 pair; this amplicon includes the target site.\",\n \"The smaller of the two amplicons, the inner amplicon, contains base pairs which lie within the bounds of the larger outer amplicon (i.e.\",\n \"is “nested” within the larger amplicon), but does not contain the target site.\",\n \"The ultimate product of the PCR reaction is double strand DNA of two lengths, only one of which (the longer) contains the target site, and the shorter of which contains a subset of the bases contained in the longer strand.\",\n \"In a preferred embodiment of the invention, the same primer functions as both the outer and inner forward primer.\",\n \"However, those of skill in the art will recognize that this need not be the case.\",\n \"With reference to FIG.\",\n \"1, a distinct forward inner primer (“P4”, shown in parentheses) might be designed to anneal between P1 and P2 and in the orientation of primer P1.Because the shorter amplicon contains bases which are identical to a portion of the longer amplicon, when the ds strands are denatured and allowed to reanneal, some (approximately 50%) of the shorter single strand DNA of the inner amplicon will hybridize with matching portions of the single strand DNA of the outer amplicon.\",\n \"This cross hybridization between the two amplicons creates two different inner-outer “hybrid” structures as depicted in FIG.\",\n \"1.In one of the two structures, the “ligatible structure” of FIG.\",\n \"1, a 3′ overhang will be present, the bases of which originated from the outer amplicon and contain the target site.\",\n \"This structure can be used, for example, for ligation based applications.\",\n \"By “ligation based applications” we mean the smaller amplicon is one of the two or more pieces of DNA that are joined together by a DNA/RNA ligase.\",\n \"In the other structure, the “sequencible structure” of FIG.\",\n \"1, a 5′ overhang will be present, the bases of which originated from the outer amplicon and also contain the target site.\",\n \"This second type of structure will be directly amenable to sequencing applications.\",\n \"By “sequencing applications” we mean that the smaller amplicon acts as a sequencing primer to sequence the DNA downstream from it.\",\n \"The sequencing can be only one base in length (i.e.\",\n \"SBE) or in the tens or in the hundreds of bases.\",\n \"Thus, in a single nested PCR reaction followed by denaturation and annealing, it is possible to generate both sequencible and ligatible structures.\",\n \"Of course, some (approximately 50%) of the single strand DNA will renature to form the original ds PCR products (“structures same as products” in FIG.\",\n \"1).\",\n \"Years of improvement in primer design and thermocycling conditions have increased the success rate for single target PCR reactions to greater than 90%.\",\n \"However, multiplex reactions such as those in a nPCR reaction, have a much lower success rate and extensive optimization is typically required, usually because it is very difficult to obtain amplified products in equal amounts.\",\n \"The usual solution is to increase the primer concentration for the weakly amplified target(s).\",\n \"This suggests that it is annealing efficiency that is responsible for the unequal amplification.\",\n \"In U.S. Pat.\",\n \"No.\",\n \"5,882,856 to Shuber, the complete contents of which is herein incorporated by reference, an alternative solution is proposed in which all primers utilized have two domains, one which is target specific as in regular PCR designs, and one which is a “common domain”.\",\n \"For a set of PCR primers to be multiplexed, two common domains are utilized.\",\n \"A first common domain is a sequence shared by all forward primers and is attached at the 5′ end of the target specific domain.\",\n \"A second common domain sequence is shared by all reverse primers and is attached at the 5′ end of the target specific domain.\",\n \"The common domain facilitates annealing of different primer sets because the common domain has a higher annealing temperature than the target specific domain.\",\n \"In the practice of the present invention, common sequence domains or “sequence tags” as taught by Shuber may be utilized for both inner and/or outer PCR primers.\",\n \"The common sequence tag for the outer PCR primers would help in multiplexing PCR so that amplification would result in the production of equal amounts of product.\",\n \"The common sequence tag for the inner primers also serves to even the levels of PCR products, and in addition may provide restriction sites that can be used to generate precise fragment ends for genotyping reactions such as sequencing and/or ligation reactions.\",\n \"The combination of incorporating special restriction enzyme recognition sequences in such a sequence tag, and the deliberate positioning of the inner PCR primers make it possible to precisely tailor the resulting PCR products for any of a variety of purposes, including but not limited to primer extension, sequencing, ligation, pyrosequencing, structure specific cleavage, and other genotyping reactions.\",\n \"Those of skill in the art will recognize that placement of the P2 primer can be anywhere between the target site and the other primer of the inner primer pair, so long as a useful PCR product is produced by PCR amplification of the inner primer pair, and the target site itself is not amplified.\",\n \"The positioning of the P2 primer relative to the target SNP may vary depending on the mechanism of the genotyping reactions.\",\n \"For example, for SBE reactions, the end of the primer should be immediately 5′ to the SNP site.\",\n \"For Multi-Target Sequencing (NulTarSeq) reactions, there may be a gap between the end and the target SNP site.\",\n \"In a preferred embodiment, for MulTarSeq reactions the gap may be from about 1 to 50 bases, or more preferably from about 5 to about 20 bases, and most preferably from about 10 to about 15 bases.\",\n \"Further, as described above, the desired length of the final product from the amplification of PCR primers containing common sequence tags may also be determined by the presence of a restriction enzyme cleavage site within the tag.\",\n \"In yet another embodiment of the present invention, the amplification reactions may be carried out in a step-wise fashion using primary and secondary primers.\",\n \"In this embodiment, a single PCR is divided into two sequential reactions, the first of which uses the primary primer set and the second of which uses the secondary primer set.\",\n \"The primary primers may be designed to have two domains (target specific and common) as described above.\",\n \"The secondary primer set contains only the common tails from the primary primers.\",\n \"For the first reaction, the primary primers are used in only limited but equal amounts, e.g.\",\n \"about 1-5% of the amount for a regular PCR i.e.\",\n \"about 0.1 to about 100 nM.\",\n \"The rationale is that by limiting the amount of primers for each set multiplexed in the reaction, the system is forced to produce limited but equal amounts of products for all amplicons.\",\n \"When the more robust primers are used up (because of the limited amount available) the system will work with the less efficient primers.\",\n \"The purpose of the primary PCR is to amplify enough templates from the genomic DNA to carry out the secondary PCR.\",\n \"Since all primary primers have limited but equal amounts, this forces the primary PCR to produce limited but near equal amounts of templates for the secondary PCR.\",\n \"For the secondary PCR, the secondary primers, consisting of only one set for all primary amplicons, is used.\",\n \"In essence this secondary reaction is a single PCR that amplifies multiple templates.\",\n \"The net result of this two-step PCR process is to compensate for differences in efficiency across PCR reactions.\",\n \"The purpose of the inner PCR in the nested reaction is to synthesize a DNA fragment whose size and position relative to the outer PCR can be precisely controlled.\",\n \"By altering the position of the P2 primer relative to P1, and by optionally including sequence tags with restriction sites, it is possible to design longer or shorter PCR fragments.\",\n \"The synthesized fragments can then be used in any of a variety of genotyping reactions, including but not limited to primer extension, sequencing, ligation, pyrosequencing, structure specific cleavage, and the like.\",\n \"For example, the sequencible structure fragments may be used as extension primers in SBE and MulTarSeq reactions.\",\n \"The direct production of sequencible structures by the methods of the present invention thus permits investigators to take advantage of the ready availability of DNA sequencers for the purposes of genotyping.\",\n \"Relatively long primers can be synthesized in this manner in a feasible and much more cost effective way than, as is typically done, by chemical synthesis.\",\n \"For example, the length of such a fragment can range from about 35 to about 1000 bps, and is preferably in the range of about 50 to about 650 bps.\",\n \"To use a standard sequencing reaction to determine the sequences surrounding a polymorphic site, it is simply necessary to add an appropriate sequencing mixture without using any sequencing primer, because the sequencible structure provides the free 3′ end for the sequence reaction.\",\n \"Those of skill in the art will recognize that several methods of sequencing are well established, and that new sequencing methodologies are continually being developed.\",\n \"In the practice of the present invention, any suitable method of sequencing may be utilized.\",\n \"Examples of such methods include but are not limited to Sanger (dideoxy)-based sequencing methods and pyrosequencing.\",\n \"The procedure illustrated in FIG.\",\n \"1 amplifies a single target and relies on multiplex PCR amplification since more than one pair of primers is used in a single reaction.\",\n \"However, those of skill in the art will recognize that in many cases it will be desirable to amplify more than one target site at a time.\",\n \"In this case, the design of the inner primer pairs for each target site may be such that an ordered series of PCR products of varying lengths is produced, i.e.\",\n \"the sequencible structure formed for each target site differs in length from the sequencible structures formed for each of the other target sites.\",\n \"(See FIG.\",\n \"2A).\",\n \"For example, an ordered series of sequencible structures from about 50 to about 500 bps in length can be designed, thus adding an additional distinguishing feature to each of the amplified target sites.\",\n \"Because of this feature, multiple sets of PCR products can be sequenced in a single reaction providing that the length of sequencing is limited so that their chain termination fragments do not overlap in a sequencing gel.\",\n \"This would allow the sequencing of multiple targets in a single reaction and analysis in a single channel (e.g.\",\n \"a lane or capillary).\",\n \"Those of skill in the art will recognize that several ways exist to control the length of reading for sequencing reactions.\",\n \"For example, the length of reading can be adjusted by changing the ratio of ddNTP/dNTP in the reaction mixture.\",\n \"For standard Sanger sequencing reactions, the ratio is typically about 1:100, a ratio intended to promote relatively long reads (e.g.\",\n \"about 600-800 bps).\",\n \"Those of skill in the art will recognize that, by systematically altering and testing the ratio, it is possible to find the optimal reading length for a given application.\",\n \"For most SNP genotyping purposes, a read of from about 1 to about 50, and preferably from about 1 to about 25 bases would be sufficient, requiring a low ddNTP/dNTP ratio of preferably in the range of about 1:0 to about 1:10, and more preferably about 1:0 to about 1:5.For other applications, the optimal length can be attained by adjusting the ratio, i.e.\",\n \"the ratio may be optimized by methods which are well known to those of skill in the art in order to achieve the desired sequence read length.\",\n \"Precise control of the read allows very high level multiplexes to be carried out (e.g.\",\n \"100 to 200×).\",\n \"In order to carry out the practice of the present invention, primer design involving placement of the primer and the optional addition of a sequence tag must be carried out.\",\n \"Further, those of skill in the art will recognize that other factors influence primer design and must be taken into account as well.\",\n \"For example, the exact sequence of bases will influence the strength of the hybridization of the primer with its cognate DNA (the sequences of based to which it binds during PCR) and thus must be taken into account when planning PCR amplification protocols; the possibility of primer-dimer formation is taken into account, as is the potential for the formation of secondary structure, etc.\",\n \"Those of skill in the art are well acquainted with such issues and numerous resources exist to aid the skilled practitioner in these various aspects of primer design.\",\n \"As discussed above, the common domains of the inner primers may be designed to include restriction enzyme recognition sites.\",\n \"The restriction enzymes preferred are those that cleave DNA strands downstream from their recognition sites and generate blunt ends products.\",\n \"For sequencing applications restriction enzymes that cleave DNA downstream and generate 3′ overhangs can also be used.\",\n \"For ligation applications restriction enzymes that cleave DNA downstream and generate 5′ overhangs can also be used.\",\n \"Examples of such sites include but are not limited to these enzymes: Mly I, Fst F51, Bpm I, Eco571, Bsg I, Eci I, Bst71 I, Alw26 I, Ksp62 I, BceA I, etc.\",\n \"The other nucleotides in the sequence of a common domain inner primer are selected to balance the melting temperature with that of the common domain of the outer primers.\",\n \"With respect to the design of common domain sequences for outer primers (i.e.\",\n \"those which do not contain restriction sites), the sequences are selected so that so it will match the melting temperature of the common domain of the inner primer.\",\n \"In general, the generation of the sequencible and ligatible structures of the present invention will be carried out as follows: a PCR amplification reaction is carried out, followed by a restriction with a suitable restriction enzyme if a restriction site was incorporated into a common sequence tag of the inner primers.\",\n \"This is followed by treatment to “clean up” the reaction e.g.\",\n \"with shrimp alkaline phosphatase (SAP) and exonuclease I (Exo I) to degrade excess PCR primers and dNTP left over from the PCR reaction, and to remove the overhanging ends of restriction digestion.\",\n \"(Alternatively, filtration via sephadex gel may also be employed.)\",\n \"After SAP/Exo I digestion, a high temperature denature step is performed.\",\n \"This denature step serves two purposes: one is to inactivate both SAP, Exo I and the restriction enzyme; the other is to completely denature double stranded DNA.\",\n \"During the cooling process that follows, the four single stranded DNA fragments from the two PCR products will form four kinds of structure.\",\n \"Two of them are the exactly same as the PCR products and are both blunt ended.\",\n \"Two new structures formed between the complementary strands of the large and small PCR products, one of them with a 5′ end single stranded, the other one with a 3′ end single stranded.\",\n \"The one with a 5′ end single stranded provides a free 3′ end for sequencing reaction/primer extension and for ligation, and is referred to as a “sequencible structure”.\",\n \"The one with single stranded 3′ end provides a free 5′ end for ligation, and is referred to as a “ligatible structure”.\",\n \"These four structures will have equal concentrations if the quantity of the two PCR products is equal.\",\n \"In this case, the sequencible and ligatible structures will represent about 50% of the reannealed products.\",\n \"This amount is sufficient for both sequencing/primer extension and ligation reactions to make new products.\",\n \"In some embodiments of the present invention, a single target site is analyzed by the methods of the present invention, and the PCR step is carried out by multiplex PCR, i.e.\",\n \"amplification with both the inner and outer primers is carried out in the same reaction.\",\n \"However, those of skill in the art will recognize that two separate PCR reactions may also be carried out, i.e.\",\n \"the inner and outer PCR reactions are carried out separately.\",\n \"In this case, the PCR products from the two reactions may be combined later at some point prior to the renaturation step so that the single strands can reanneal.\",\n \"Similarly, in other embodiments of the present invention, it may be desirable to PCR amplify more than one target site at a time.\",\n \"In some cases, this may be carried out by a multiplex PCR reaction in which inner and outer primers for all target sites of interest are amplified simultaneously in a single reaction.\",\n \"Alternatively, one or more sites may be amplified together, or each site may be amplified alone, either in a single reaction with both inner and outer primers, or in two reactions, one for the inner and one for the outer primers as described above.\",\n \"Any combination of PCR amplification reactions may be utilized in the practice of the present invention, so long as the procedure results in the production of suitable sequencible and/or ligatible structures.\",\n \"In a preferred embodiment, the present invention provides methods which ultimately utilize DNA sequencers or the like to detect the products formed from sequencible and/or ligatible structures created by the present methods.\",\n \"Those of skill in the art will recognize that many suitable techniques exist by which the sequencible products can be produced for analysis by a DNA sequencer.\",\n \"Examples include but are not limited to Sanger-type sequencing reactions, single base extension (SBE), and pyrosequencing.\",\n \"Ligatible structures may be analyzed directly on a DNA sequencer after a labeled oligonucleotide (e.g.\",\n \"fluorescently labeled) is joined to the 5′ phosphate of the ligatible structure via a ligation reaction.\",\n \"Those of skill in the art are well acquainted with Sanger-type sequencing reactions, which are based on the interruption of enzymatic extension of the DNA chain at the 3′ hydroxyl via incorporation of 2′-3′-dideoxy analogs.\",\n \"Four sets (one for each dideoxy analog) of such chain-terminated fragments of differing lengths are generated and “read” (detected) by a DNA sequencer to yield a sequence.\",\n \"Typically, the fragments are labeled with radioactivity or fluorescent tags for detection.\",\n \"Single base extension is essentially a sequencing reaction, the difference being that SBE uses only dye-terminators, resulting in the extension of the primer by only a single base.\",\n \"As described above, in the practice of the present invention, the length and position of the extension primer relative to the polymorphic site may be controlled so that it is possible to create a primer which anneals immediately upstream of a polymorphic site.\",\n \"Then, in a regular SBE reaction, the polymorphic base can be determined by identifying which base(s) is incorporated into the primer.\",\n \"If several target sites are to be analyzed, a set of primers can be designed which, upon amplification of the targeted DNA, produce an ordered set of such sequencible structures of varying, non-overlapping lengths.\",\n \"When such structures are extended by dye-labeled ddNTPs, an ordered series of SBE products of varying, non-overlapping lengths is produced.\",\n \"In the proposed nested PCR design, because extension primers from about 50 to over 500 bases can be obtained, it is possible to separate these SBE products in a sequencing gel/capillary.\",\n \"A regular sequencing gel can sequence about 500-800 bases.\",\n \"Therefore, over 100 SNPs can be analyzed in a single channel by loading their corresponding SBE products in one lane/capillary when the extension primers are designed to be positioned, for example, every 4-5 bases apart for a full range of 500-600 bases.\",\n \"In order to successfully carry out SBE reactions, the ends of the primers must be precise, i.e.\",\n \"it is necessary to control precisely where the inner amplicon ends.\",\n \"The use of Taq DNA polymerase may in some cases be disadvantageous due to its known extendase activity which interferes with such precision.\",\n \"Those of skill in the art will recognize that a variety of means are available to overcome such a problem.\",\n \"For example, the use of high fidelity polymerases may be employed.\",\n \"Pfu DNA polymerase is a well characterized DNA polymerase that has been shown to have high fidelity of replication (Cline 1996).\",\n \"It is known that Pfu DNA polymerase does not add extra A's at the 3′ end (Cline 1996; Hu 1993).\",\n \"Based on a study by Cline et al (Cline 1996) the use of Pfu DNA polymerase gives precise ends of inner PCR products.\",\n \"A second alternative is to use ribonucleotides and/or abasic analogs in the inner primer to control DNA polymerases' end point.\",\n \"Some DNA polymerases do not have reverse transcriptase activity, Pfu and Vent polymerases are examples (Perler 1996).\",\n \"These enzymes can not incorporate any nucleotide when they encounter a ribonucleotide base in a template.\",\n \"Coljee et al recently reported a strategy to create precise overhang at the ends of PCR products by using ribonucleotides (Coljee 2000).\",\n \"The strategy inspired us to use same mechanism to control the ends of inner PCR products.\",\n \"Thus, inner primers may be designed to overlap the target SNP site, at the SNP site, instead of using regular deoxyribonucleotide, a ribonucleotide is used.\",\n \"DNA polymerases can use a chimeric oligonucleotide as primer efficiently (Gal 2000; Stump 1999), so the use of chimeric primers would not compromise PCR efficiency.\",\n \"When the complementary strands are synthesized, the DNA polymerases will stop one base before the ribonucleotide base in the chimeric inner primers, creating very accurate and precise ends for SBE.\",\n \"It is also possible to create chimeric primers containing several ribonucleotide bases to create several base overhang for the purpose of ligation (Coljee 2000).\",\n \"Similarly, a nucleotide analog that does not contain the base, the abasic analog, has similar effects on DNA polymerase (Gal 2000; Gal 1999) and may be used.\",\n \"Yet another alternative is to use restriction enzymes to produce precise 3′ ends for sequencible structures.\",\n \"There are some restriction enzymes that cleave DNA strands downstream from their recognition sites, e.g.\",\n \"KSP632 I, Alw26 I, and Bst71 I.\",\n \"As discussed above, to take advantage of these features, a common domain (sequence tag) may be added to the 5′ end of the inner PCR primers.\",\n \"The common domains contain restriction enzyme sites which can be positioned so that the 3′ extending ends are located just before the polymorphic sites after the tail is cleaved off.\",\n \"The use of a common domain for inner primers could also help to even the amounts among different amplicons when multiplexed.\",\n \"Pyrosequencing is a unique technique that incorporates a sequencing reaction and real time detection together.\",\n \"Since, as in a regular sequencing reaction, pyrosequencing uses DNA polymerase, the free 3′ termini in the sequencible structures can function as extension primers.\",\n \"Since pyrosequencing is a real time system, it is not amenable to the analysis of multiplex reactions.\",\n \"There would thus be no need to use a common tail for the inner PCR primers.\",\n \"The primers for a reaction whose products are destined for analysis by pyrosequencing are designed to accommodate the needs of pyrosequencing, preferably from about 5 to about 8 bases upstream of the target polymorphic site.\",\n \"The ligatible structures produced by the methods of the present invention have a free 5′ phosphate.\",\n \"If a common tag is included in the design, when the common tag is removed from the inner PCR products, restriction enzyme digestion leaves the 5′ phosphate on the ligatible structure.\",\n \"To make such a ligation site at a polymorphic site, the cleavage site of restriction enzymes used in the inner PCR primer must be immediately down stream of the polymorphic site on the complementary strand.\",\n \"Those of skill in the art will recognize that, because all of these restriction enzymes produce a 5′ over hanging strand, a suitable design of restriction site for ligation assay would not be suitable for SBE reaction, but could be used for MulTarSeq reaction.\",\n \"Further, one advantage of the use of ligation to score a polymorphism is that it can be combined directly with the PCR reaction; resulting in a closed tube assay.\",\n \"To be detected by a DNA sequencer, DNA fragments have to be labeled with fluorescence dyes.\",\n \"In the present nested PCR design, the PCR products are not labeled with any fluorescence dye.\",\n \"Thus, the ligatible structures can be detected only if they are ligated to a labeled oligonucleotide, e.g.\",\n \"one that bears a fluorescent label.\",\n \"To be ligated onto a ligatible structure created by restriction enzyme digestion and subsequently detected by a DNA sequencer, an oligonucleotide would have a 3′ OH group and 5′ fluorescence labeling.\",\n \"Current DNA sequencers typically analyze fluorescence with a spectrum between 510-610 nm.\",\n \"Those of skill in the art will recognize that there are many fluorescence dyes available in this range.\",\n \"For example, the BODIPY series of fluorescent dyes have been shown to minimize emission overlaps between dyes (Metzker 1996).\",\n \"Examples of other dyes which may be utilized to carry out this aspect of the invention include but are not limited to FAM, fluorescein, R110, R6G, TAMRA, ROX, Texas red, etc.\",\n \"If only a single locus is of interest is to be analyzed, locus-specific fluorescence-labeled oligos may be utilized.\",\n \"However, in order to carry out high level multiplex reactions, sets of degenerate, fluorescence-labeled oligos are desirable.\",\n \"For example, in order to make ligatible structures suitable for high throughput genotyping assays, a common set of four fluorescence dye labeled degenerate oligonucleotides may be utilized.\",\n \"The degeneration of the oligonucleotides is to make them reusable for all potential assays.\",\n \"If an oligonucleotide with a specific sequence is used, it would only work for one particular target.\",\n \"Several important issues must be taken into consideration regarding the design of the degenerate oligonucleotides.\",\n \"One is the length of the oligonucleotide, which effects both ligation efficiency and the concentration of a specific sequence in the degenerate oligo population).\",\n \"For example, for a 6-7 mer oligo containing 5-6 degenerate bases, any given sequence would have a concentration of ¼3-¼4 (i.e.\",\n \"1/1024- 1/4096).\",\n \"If the total concentration for a 7-mer degenerated oligo used in ligation is 1 μM, the effective concentration for any giving sequence would be greater than 0.2 nM, a concentration at least 100 fold above the detection limit of sequencing gel/capillary.\",\n \"Those of skill in the art will recognize that the optimal length of the degenerated oligo and its concentration may vary from experiment to experiment, depending on the level of degeneracy and multiplex used in the reaction.\",\n \"However, the oligo will generally be in the range of about 5- about 20 bps in length with from about 4 to about 19 degenerate bases, and more preferably in the range of about 6 to about 8 bps in length with about 4 to about 6 degenerate bases.\",\n \"Another consideration is the melting temperature for the degenerated oligonucleotide.\",\n \"As is well known to those of skill in the art, the Tm will vary depending on factors such as the GC content, the length of the oligo, and the like.\",\n \"For example, increasing the GC content and/or the length of an oligo increases its Tm.\",\n \"Increasing the length of oligonucleotide will increase Tm, but would decrease the effective concentration for a specific sequence.\",\n \"In addition, other factors such as the ionic strength of the ligation buffer also affect Tm significantly.\",\n \"Other considerations involved in oligo design which are well-known to those of skill in the art include the number and position of bases which are degenerated.\",\n \"The number of degenerated bases will have a significant impact on the effective concentration and Tm.\",\n \"For example, if 5 degenerated bases are used within a 7 mer, 16 oligos must be synthesized, but if 6 degenerated bases are used, only 4 oligos are necessary.\",\n \"While one specified position must match the polymorphic site if multiple specified positions are used, the position does not necessarily have to be the last one on the 3′ end.\",\n \"Further, the position of the ligation site relative to SNP site (the 5′ end or the 3′ end) may also be altered to optimize a given experiment.\",\n \"Previous studies (Tong 2000; Tong 1999) have suggested that DNA ligase exhibits better allelic discrimination when the ligation site is on the 3′ end.)\",\n \"Those of skill in the art will recognize that many tools exist and are well-known to the skilled practitioner for optimizing such parameters, thereby facilitating the design of suitable oligos.\",\n \"(See, for example, Delahuntyl996, Tong 2000; Tong 1999).\",\n \"Examples include but are not limited to various computer soft ware packages such as DNA Star, Oligo, Primer Express, etc.\",\n \"Other factors which may be considered include the type and concentration of ligase to be employed.\",\n \"For example, a 6-7 mer oligo with a moderate GC content should exhibit a melting temperature between 3040° C., a Tm sufficient to perform a cyclic ligation assay.\",\n \"The use of thermal stable DNA ligase would provide an opportunity for multiple cycling of a ligation assay at this temperature, which would compensate for lowered efficiency of the reaction.\",\n \"Those of skill in the art will recognize that a plethora of protocol references, kits, etc.\",\n \"are readily available to facilitate carrying out such manipulations of the DNA.\",\n \"The precise nature of the protocols which are used is not a central feature of the present invention, and any of many suitable means for carrying out the various steps of PCR amplification and detection and analysis of the amplicons may be utilized in the practice of the present invention.\",\n \"Those of skill in the art will recognize that a variety of additional well-known techniques may be reliably utilized in the practice of the present invention, examples of which include but are not limited to: mass spectroscopy, ELISA techniques, fluorescence intensity/energy transfer/polarization, various micro assays (e.g.\",\n \"DNA/RNA arrays), microfluidic devices, and magnetic/color-coded micro beads.\",\n \"In a preferred embodiment of the present invention, the method of detection is capillary electrophoresis.\",\n \"The demand from the human genome project greatly stimulated technology development in DNA sequencing and, in return, improvements in DNA sequencing greatly speeded up the human genome project.\",\n \"As a result, capillary DNA sequencers became widely available to researchers in both academe and industry.\",\n \"Capillary DNA sequencers are highly sensitive and automated detection systems.\",\n \"They also have a relatively broad detection range, covering at least three orders of magnitude.\",\n \"Their analytical power is better than most signal detection systems currently used for SNP genotyping.\",\n \"Capillary DNA sequencing systems have been proven to deliver ultra high throughput in many DNA sequencing centers around the world.\",\n \"Capillary electrophoresis has been used in DNA polymorphism analysis (Barta, 2001; Medintz 2001).\",\n \"Most of these studies exploited the sensitivity and speed of separation.\",\n \"Lindblad-Toh et al (2000) used a DNA sequencer to increase throughput.\",\n \"In this study, the authors chemically synthesized oligonucleotides from 18-50 bases as extension primers and succeeded in scoring 14 SNPs in one lane (The size difference between primers was 4 bases, there were a few sets of oligos of the same length but of different sequences that were used to score two different SNPs by different colors).\",\n \"This study essentially proved that DNA sequencers had the potential for high capacity SNP scoring using the SBE format, and the length of primers was the limiting factor for throughput.\",\n \"The wide availability, high detection sensitivity, high throughput capacity and automation readiness of capillary DNA sequencers call for high capacity applications, and SNP genotyping is one of such applications.\",\n \"If these two are united, considerable advances in cost and throughput would be achieved.\",\n \"Many did not consider DNA sequencers for high throughput SNP scoring mainly because it was not cost-effective.\",\n \"DNA sequencers separate DNA fragments by sizes and colors.\",\n \"In order to separate genotyping products by a sequencer, the products must be different in sizes and or colors.\",\n \"To separate two SBE products, for example, SBE primers must be different in length or the extended bases must be different in color.\",\n \"Otherwise, the two products of same length and same colors would not be distinguished.\",\n \"While it is reasonable to chemically synthesize primers of 15-40 bases, this size range gives only limited multiplex capacity (Lindblad-Toh 2000)).\",\n \"To chemically synthesize primers longer than 50 bases is not economical, especially in high throughput settings.\",\n \"But a sequencer can separate DNA fragments up to 600-800 bases.\",\n \"In the practice of the present invention as described above, SBE products can be arranged to be 5 bases apart for 600-800 bases, allowing the loading over 100 SBE products in one lane.\",\n \"Multiplexing at this level makes it reasonable to use sequencers for SNP scoring.\",\n \"Furthermore, practical protocols for using DNA sequencers are well-established and many handling and procedures involved have been automated and are readily available for large scale applications.\",\n \"The following examples are intended to be illustrative in nature and are not intended to be limit the scope of the invention in any way.\",\n \"EXAMPLES Example 1 Formation of Sequencible Structures Sets of nPCR primers were designed as described to amplify DNA fragments from six markers of human chromosome 6 and individual nPCR was performed using Taq-Gold DNA polymerases.\",\n \"Clean-up digestion was carried out using shrimp alkaline phosphatase and E. coli exonuclease I (37° C. 1 hour) After the clean-up digestion, a high temperature denaturing (95° C. for 15 min.)\",\n \"was performed and the samples were allowed to cool to room temperature.\",\n \"A sequencing reaction was performed using standard sequencing kits but without sequencing primers.\",\n \"Sequencing reactions were performed either: 1) under standard cyclic sequencing program: 25 cycles of 96° C. for 1 min., 50° C. for 15 sec.\",\n \"and 60° C. for 4 min; or 2) using this condition: 25 cycles of 96° C. for 1 min., 80° C. for 4 min.\",\n \"The purpose of these experiments was to test whether a sequencible structure formed after denaturing and reannealing of the outer and inner PCR products together.\",\n \"If we obtained sequences and the sequences matched the inner PCR primer position, this would confirm that sequencible structures did form as expected.\",\n \"The results from experiments with two of the six markers are shown in FIGS.\",\n \"3A and B and FIGS.\",\n \"4A and B.\",\n \"These figures show the results obtained with the second sequencing reaction protocol.\",\n \"FIGS.\",\n \"3A and 4A show PCR primer positions and orientations, and the expected sequences.\",\n \"Note that in FIG.\",\n \"4, the inner PCR primer is designed in the same orientation as forward, so the sequence reading from a sequencing reaction would be reverse complementary to the sequences highlighted with italics.\",\n \"FIGS.\",\n \"3B and 4B show sequencing reads from the ABI 377 sequencer analysis software.\",\n \"For the 6 markers tested, we obtained sequences for five, and all of these sequences matched the expected sequences exactly.\",\n \"Full length sequence reads (from the ends of inner PCR products to the ends of outer PCR products) were obtained when the standard cyclic sequencing conditions were used (data not shown).\",\n \"Much shorter sequence readings (30-50bases) were obtained when the extension temperature was changed to 80° C. (as in FIGS.\",\n \"3 and 4).\",\n \"For 4 of the 5 makers that produced sequences the sequence started right after the inner primers, one started 3 bases away from the end of the primer.\",\n \"For both sequencing conditions, the starting positions of sequencing were the same.\",\n \"Example 2 Creating Sequencible Structures with Products from Extendase-Free Pfu DNA Polymerases that can be Used for SBE Genotyping Reactions The variations of starting position of the obtained sequences (Example 1) suggested that the positioning of inner PCR primers alone may not be sufficient to obtain sequencible structures with precise extending ends.\",\n \"An alternative is the use of extendase-free DNA polymerases that do not add the 3′ A base at the ends of PCR products.\",\n \"Thus, experiments were carried out using Pfu DNA polymerases which have been reported for lacking the extendase activity (10, 28).\",\n \"Inner PCR primers were designed so that their products would be positioned right before the polymorphic site if the products formed sequencible structures.\",\n \"Inner and outer PCR reactions were performed separately using Pfu DNA polymerases and the products wqere pooled together to form the sequencible structures.\",\n \"Then a SBE reaction was performed using SNaPshot kit from Applied BioSystems.\",\n \"SBE reactions were purified and analyzed by ABI 377 DNA sequencer using GeneScan.\",\n \"Three samples were shown in FIG.\",\n \"5, representing three genotypes for the marker: Homozygous A/A (upper panel), heterozygous A/G (middle panel) and homozygous G/G (lower panel).\",\n \"The experiments demonstrated that Pfu DNA polymerases produce inner PCR products with precise ends that can be used to form sequencible structures for SBE reactions and to score genotypes correctly.\",\n \"In a parallel experiment using Taq DNA polymerases multiple products were observed and genotypes were not clear (data not shown).\",\n \"The results clearly show the differences between the two DNA polymerases.\",\n \"Example 3 Control of the Reading Length for Multiple Target Sequencing The reading length of sequencing reactions is principally determined by the ratio of dNTPs/ddNTPs.\",\n \"For typical sequencing applications, long sequence reads are obtained by using a high dNTPs/ddNTPs ratio (100:1) provided in commercial sequencing kits.\",\n \"In the practice of the present invention, a short stretch of DNA surrounding the SNP targets will be sequenced.\",\n \"For this purpose it is desirable to control the length of sequence reads so that multiple sequencing reactions can be loaded in one lane/capillary without overlapping each other.\",\n \"Currently there is no commercially available kit for this purpose, but two products, the SNaPShot from Applied Biosystems and the SNuPe from Amershan Biosciences, have the potential to be modified for this purpose.\",\n \"Both these kits were designed for SBE reactions, containing only dye-labeled ddNTPs.\",\n \"The concentration of nucleotides in the mix is proprietary.\",\n \"Individually available dye-labeled nucleotides were used to assemble a testing kit to demonstrate the principle that reading length can be controlled by dNTPs/ddNTPs ratio.\",\n \"FIG.\",\n \"6 shows an example of the sequencing reaction.\",\n \"In this reaction, dye-labeled acyclo-ddNTPs were used to assemble the mix.\",\n \"The sequencing reaction was performed using pGEM plasmid and M13-21 primer.\",\n \"The ratio of dNTPs/ddNTPs and thermal cycling conditions are detailed in the figure legend.\",\n \"The results showed a clean read of 30-40 bases after the primer.\",\n \"This example demonstrated that the reading length can be effectively controlled by the ratio of dNTPs/dDNTPs.\",\n \"To practice the current invention, especially the MulTarSeq, a specially formulated sequencing mix is therefore desirable.\",\n \"Example 4 Comparison of Two-Step PCR Amplification Using Primary and Secondary Primer Sets Four SNP markers randomly were chosen from our SNP collection and PCR and nPCR primers were designed.\",\n \"Both outer and inner primers had two designs, a simple design and the two domain design.\",\n \"The sequences used for the common domains of the two domain were the M13 universal forward primer: cccagtcacgacgttgtaaaacg (for forward outer PCR primers) and M13 universal reverse primer: agcggataacaatttcacacagg (for reverse outer PCR primers).\",\n \"For the inner PCR primer the common domain sequence was tcaGCAGCatGTCTCttcca, which contains recognition sites for two restriction enzymes (Bst71 I and Alw26 I, in capital letters).\",\n \"Multiplex PCR and multiplex nPCR reactions were performed to compare the primer designs and protocols.\",\n \"PCR products were then analyzed by agarose gel electrophoresis.\",\n \"FIG.\",\n \"7 showed the results.\",\n \"Lane 1 of FIG.\",\n \"7 represents a 4× multiplex PCR using simple primer design.\",\n \"As ca be seen, only two bands are visible (480,510 bp).\",\n \"They could not be resolved by agarose gel electrophoresis.\",\n \"Lane 2 represents the same 4 amplicons, but generated using the two domain design.\",\n \"In this case, the two smaller products (400, 430 bp) are clearly seen.\",\n \"Note the size shifted for the larger products due to extra length (46 bp) in the two domain primers.\",\n \"Lane 3 represents 4× multiplex NPCR with the same amplicons but carried out by the two-step porcedure.\",\n \"In addition to the 4 outer PCR products, 4 inner PCR productscan also be seen.\",\n \"This experiment demonstrates that the two domain design significantly improves both multiplex PCR and multiplex NPCR, and that the two-step procedure is especially effective to improve multiplex nPCR.\",\n \"Example 5 Use of Multiplex nPCR to Create an Ordered Series of Sequencible Structures This example is provided to illustrate how a series of sequencible structures are formed from two multiplexed PCRS.\",\n \"A set of five SNPs are selected and NPCR primers are designed using the two domain design.\",\n \"In order to run all products on a single sequencing lane/capillary, the inner PCR products are arranged to span 150 to 400 bps range, 50 bps apart from one another.\",\n \"The common domain for the inner primers should contain a restriction enzyme recognition site to produce blunt ends after the amplification.\",\n \"Two 5× mulitiplexing PCRs are performed separately: one uses only the outer PCR primers the other uses only the inner PCR primers.\",\n \"After the amplification an appropriate restriction enzyme is used to remove the common tag sequences from the inner end of the inner PCR products.\",\n \"The outer products and the inner products then are combined, purified, denature and reannealed together.\",\n \"A single base extension reaction is performed using the SNaPshot kit from Applied Biosystems.\",\n \"The products then are purified and run on DNA sequencer.\",\n \"The analysis of the extension products will identify the genotypes of the sample.\",\n \"While the invention has been described in terms of its preferred embodiments, those skilled in the art will recognize that the invention can be practiced with modification within the spirit and scope of the appended claims.\",\n \"Accordingly, the present invention should not be limited to the embodiments as described above, but should further include all modifications and equivalents thereof within the spirit and scope of the description provided herein.\",\n \"REFERENCES Armstrong B, Stewart M, Mazumder A.\",\n \"2000.Suspension arrays for high throughput, multiplexed single nucleotide polymorphism genotyping.\",\n \"Cytometry 40:102-8 Barta C, Ronai Z, Sasvari-Szekely M, Guttman A.\",\n \"2001.Rapid single nucleotide polymorphism analysis by primer extension and capillary electrophoresis using polyvinyl pyrrolidone matrix.\",\n \"Electrophoresis 22:779-82 Bray M S, Boerwinkle E, Doris P A.\",\n \"2001.High-throughput multiplex SNP 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\"18:760-3 Ye F, Li M S, Taylor J D, Nguyen Q, Colton H M, Casey W M, Wagner M, Weiner M P, Chen J.\",\n \"2001.Fluorescent microsphere-based readout technology for multiplexed human single nucleotide polymorphism analysis and bacterial identification.\",\n \"Hum.\",\n \"Mutat.\",\n \"17:305-16\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476970"}}},{"rowIdx":989,"cells":{"text":{"kind":"list like","value":[["Company board data processing system and method","A company board data processing system and method provides a user interface platform to supply information related to members of boards of companies to enable comparison of board members or boards at board member level.","Parameters related to board members of a plurality of boards are stored.","A used interface is generated which comprises a display of the parameters related to board members for a plurality of boards and/or individuals board members to allow comparison of board members and/or boards at boards member level.","The system also allows the user to define a group of boards and/or board members, wherein processed parameters for said group can be used for comparison purposes."],["1.A company board data processing system comprising: a board member store storing parameters related to board members of a plurality of boards organised into boards; and a user interface generator for generating a user interface displaying said parameters related to board members for a plurality of boards and/or individual board members to allow comparison of boards, at board member level, and/or board members.","2-197.","(canceled) 198.A company board data processing system as in claim 1, wherein said user interface generator is adapted to display said parameters for each member of at least one board and/or an average value of one or more of said parameters of said members of at least one board.","199.A company board data processing system as in claim 1, further comprising a board parameter processor for processing parameters for a group of boards or board members to determine processed parameters comprising at least one of one or more parameter totals, one or more parameter averages and one or more parameter deviations, wherein said user interface generator is adapted to generate a user interface to display at least one of said processed parameters for at least one group of boards or board members to allow comparison with at least one board or board member.","200.A company board data processing system as in claim 1, wherein said user interface generator is adapted to generate said user interface to display processed parameters comprising at least one of one or more totals, averages and deviations of said parameters related to a group of board members or boards to allow comparison with at least one board or board member.","201.A company Board data processing system as in claim 1, including a ranking processor for ranking members of a group of boards and/or board members based on relative value of said parameters; wherein said user interface generator is adapted to generate a user interface to display said parameters of said boards and/or board members of said group to indicate ranking.","202.A company Board data processing system as in claim 199, including a ranking processor for ranking groups of boards and/or board members based on relative value of said processed parameters; wherein said user interface generator is adapted to generate a user interface to display said processed parameters of said groups of boards and/or board members to indicate ranking.","203.A company data processing system as in claim 199, wherein said group of board members comprises a user selected and/or pre-defined group of boards or board members assigned as a peer group.","204.A company board data processing system as in claim 199, further comprising: user selector means for receiving a user selection of at least one board and/or board member for inclusion in a peer group; wherein said group of boards or board members comprises a user selected group of boards or board members assigned as a peer group; said parameter processor is adapted to process parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; and said user interface generator is adapted to generate a user interface displaying at least one of said processed peer group parameters.","205.A company board data processing system as in claim 204, wherein said user selector means is arranged to receive a user selection of at least one board for comparison with at least one board peer group, and said user interface generator is adapted to generate a user interface to display the parameters related to members of said at least one selected board, to allow the or each board to be the subject of analysis.","206.A company board data processing system as in claim 204, wherein said user interface generator is adapted to generate a user interface to display the parameters related to individual members of said at least one selected board.","207.A company board data processing system as in claim 204, wherein said user selector means is arranged to receive a user selection of at least one board member for comparison with said board member peer group, and said user interface generator is adapted to generate a user interface to display the parameters related to said at least one selected board member, to allow said at least one selected board member to be the subject of analysis.","208.A company board data processing system as in claim 204, further comprising a peer group data store storing data for one or more user selected peer groups; wherein said user interface generator is adapted to generate a user interface displaying a list of existing peer groups based on said stored peer group data; said user selector means is arranged to receive a user selection of one or more peer groups from said list; and said user interface generator is adapted to generate a user interface displaying at least one of said processed parameters for at least one of said selected peer groups.","209.A company board data processing system as in claim 204, further comprising a comparison group amendment means allowing a user to amend the constituents of a peer group.","210.A company board data processing system as in claim 209, wherein said peer group amendment means is adapted to allow a user to copy one or more constituents of a peer group to another peer group.","211.A company board data processing system as in claim 204, further comprising a searching means arranged to allow a user to search for a particular board in the board member store by inputting a specific company or board member name.","212.A company board data processing system as in claim 211, wherein said searching means is adapted to allow a user to search for a particular board in said board member store by selecting one or more parameters from a pre-defined list.","213.A company board data processing system as in claim 1, further comprising: a store storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; wherein said user interface generator is adapted to generate a user interface displaying a list of said pre-defined peer groups; the system further comprising user selector means for receiving a user selection of one or more said pre-defined peer groups from said list; wherein said user interface generator is adapted to generate a user interface displaying at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.","214.A company board data processing system as in claim 213, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.","215.A company board data processing method comprising: storing parameters related to board members of a plurality of boards organised into boards; and generating a user interface displaying said parameters related to board members for a plurality of boards and/or individual board members to allow comparison of boards, at board member level, and/or board members.","216.A company board data processing method as in claim 215, wherein said user interface displays said parameters for each member of at least one board and/or an average value of one or more of said parameters of said members of said at least one board.","217.A company board data processing method as in claim 215, further comprising processing parameters for a group of boards or board members to determine at least one of one or more parameter totals, one or more parameter averages and one or more parameter deviations, wherein said user interface displays at least one of said processed parameters for at least one group of boards or board members to allow comparison with at least one board or board member.","218.A company board data processing method as in claim 215, wherein said user interface displays at least one of one or more totals, averages and deviations of said parameters related to a group of board members or boards of to allow comparison with at least one board or board member.","219.A company Board data processing method as in claim 215, including ranking members of a group of boards and/or board members based on relative value of said parameters; wherein generating a user interface to display said parameters of said boards and/or board members of said group to indicate ranking.","220.A company Board data processing method as in claim 217, including ranking groups of boards and/or board members based on relative value of said processed parameters; wherein generating a user interface to display said processed parameters of said groups of boards and/or board members to indicate ranking.","221.A company data processing method as in claim 217, wherein said group of boards or board members comprise a user selected and/or pre-defined group of boards or board members assigned as a peer group.","222.A company board data processing method as in claim 217, further comprising: receiving a user selection of at least one board and/or board member for inclusion in a peer group; wherein said group of boards or board members comprises a user selected group of boards or board members assigned as a peer group; parameters for constituents of a peer group are processed to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; and said user interface displays at least one of said processed peer group parameters.","223.A company board data processing method as in claim 222, further comprising receiving a user selection of at least one board for comparison with at least one board peer group, and said user interface displays the parameters related to members of said at least one selected board, to allow the board to be the subject of analysis.","224.A company board data processing method as in claim 222, wherein said user interface displays the parameters related to individual members of said at least one selected board.","225.A company board data processing method as in claim 222, further comprising receiving a user selection of at least one board member for comparison with said board member peer group, and said user interface displays the parameters related to said at least one selected board member, to allow said at least one selected board member to be the subject of analysis.","226.A company board data processing method as in claim 222, further comprising storing data for one or more user selected peer groups; wherein said user interface displays a list of existing peer groups based on said stored peer group data; a user selection of one or more peer groups is received from said list; and said user interface displays at least one of said processed parameters for at least one of said selected peer groups.","227.A company board data processing method as in claim 222, further comprising receiving a user amendment to the constituents of a peer group.","228.A company board data processing method as in claim 227, further comprising receiving a user instruction to copy one or more constituents of a peer group to another peer group.","229.A company board data processing method of claim 222, further comprising receiving a specific company or board member name as an input from a user, and searching for a particular board in the board member store.","230.A company board data processing method as in claim 229, further comprising receiving a user selection of one or more parameters from a pre-defined list, and searching for a particular board in said board member store.","231.A company board data processing method as in claim 215, further comprising storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; wherein said user interface displays a list of said pre-defined peer groups; further comprising receiving a user selection of one or more said pre-defined peer groups from said list; and said user interface displays at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.","232.A company board data processing method as in claim 231, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.","233.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 215.234.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 215.235.A company board data processing system comprising: a board member store storing parameters related to board members of a plurality of boards; user selector means for receiving a user selection of at least one board and/or board member for inclusion in a peer group; a parameter processor for processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; and output means for outputting at least one of the processed peer group parameters.","236.A company board data processing system as in claim 235, wherein said user selector means is arranged to receive a user selection of at least one board for comparison with at least one of the board peer groups, and said output means is adapted to output the parameters related to members of said at least one selected board, to allow the or each board to be the subject of analysis.","237.A company board data processing system as in claim 235, wherein said output means is adapted to output the parameters related to individual members of the or each selected board.","238.A company board data processing system as in claim 235, wherein said user selector means is arranged to receive a user selection of at least one board member for comparison with the board member peer group, and said output means is adapted to output the parameters related to said at least one board member, to allow the or each board members to be the subject of analysis.","239.A company board data processing system as in claim 235, further comprising a peer group data store storing data for one or more user selected peer groups; wherein said output means is adapted to output a list of existing peer groups based on said stored peer group data; said user selector means is arranged to receive a user selection of one or more peer groups from said list; and said output means is adapted to output at least one of said processed parameters for the selected peer group.","240.A company board data processing system as in claim 235, further comprising a comparison group amendment means allowing a user to amend the constituents of said peer groups.","241.A company board data processing system as in claim 240, wherein said peer group amendment means is adapted to allow a user to copy one or more constituents of a peer group to another peer group.","242.A company board data processing system as in claim 235, further comprising a searching means arranged to allow a user to search for a particular board and/or board member in said board member store by inputting a specific company or board member name.","243.A company board data processing system as in claim 242, wherein said searching means is adapted to allow a user to search for a particular board and/or board member in said board member store by selecting one or more parameters from a pre-defined list.","244.A company board data processing system as in claim 235, further comprising: a store storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; wherein said output means is adapted to output a list of said pre-defined peer groups; said user selector means is arranged to receive a user selection of one or more said pre-defined peer groups from said list; and said output means is adapted to output at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.","245.A company board data processing system as in claim 244, wherein said stored pre-defined peer group related parameters relate to at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.","246.A company board data processing method comprising: storing parameters related to board members of a plurality of boards organised into boards; receiving a user selection of at least one board and/or board member for inclusion in a peer group; processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; and outputting at least one of said processed peer group parameters.","247.A company board data processing method as in claim 246, further comprising receiving a user selection of one or more boards for comparison with at least one of said board related peer groups, wherein the parameters related to members of said board are output, to allow the board to be the subject of analysis.","248.A company board data processing method as in claim 246, wherein the parameters related to individual members of the or each selected board are output.","249.A company board data processing method as in claim 246, further comprising receiving a user selection of one or more board members for comparison with said board member related peer group, wherein the parameters related to said board members are output, to allow the board members to be the subject of analysis.","250.A company board data processing method as in any one of claim 246, further comprising storing data for one or more user selected peer groups; outputting a list of existing peer groups based on said stored peer group data; receiving a user selection of one or more peer groups from said list; and outputting at least one of said processed parameters for the selected peer group.","251.A company board data processing method as in claim 246, further comprising receiving a user amendment to the constituents of said peer groups.","252.A company board data processing method as in claim 251, further comprising receiving a user instruction to copy one or more constituents of a peer group to another peer group.","253.A company board data processing method as in claim 246, further comprising receiving a specific company or board member name as an input from a user, and searching for a particular board and/or board member in said board member store.","254.A company board data processing method as in claim 253, further comprising receiving a user selection of one or more parameters from a pre-defined list, and searching for a particular board and/or board member in said board member store.","255.A company board data processing method as in claim 246, further comprising storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; outputting a list of said pre-defined peer groups; receiving a user selection of one or more said pre-defined peer groups from said list; and outputting at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.","256.A company board data processing method as in claim 255, wherein said stored pre-defined peer group related parameters relate to at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.","257.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 246.258.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 246.259.A company board data processing system comprising: a store storing pre-defined groups of boards and/or board members comprising peer groups and pre-defined parameters related to said peer groups; output means for outputting a list of said peer groups; and a user selector means for receiving a user selection of one or more of said peer groups from said list; wherein said output means is adapted to output at least one of said pre-defined parameters for the or each selected peer groups.","260.A company board data processing system as in claim 259, wherein said pre-defined parameters relate to at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.","261.A company board data processing system as in claim 259, further comprising a parameter processor for processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and/or one or more peer group parameter deviations; wherein said output means is adapted to output any of said processed parameters.","262.A company board data processing system as in claim 259, wherein said store stores parameters related to board members of a plurality of boards organised into boards; said user selector means is arranged to receive a user selection of one or more said boards for comparison with any said board peer group; and said output means is adapted to output the board member parameters for each member of said board individually, to allow the board to be the subject of analysis.","263.A company board data processing system as in claim 259, wherein said user selector means is arranged to receive a user selection of at least one board member for comparison with any said board member peer group, said output means is adapted to output the parameters of said at least one board member, to allow the or each board member to be the subject of analysis.","264.A company board data processing system as in claim 259, further comprising a comparison group amendment means allowing a user to amend the constituents of said peer groups.","265.A company board data processing system as in claim 264, wherein said peer group amendment means is adapted to allow a user to copy one or more constituents of one said peer group to another said peer group.","266.A company board data processing method comprising; storing pre-defined groups of boards and/or board members comprising peer groups and pre-defined parameters related to said peer groups; outputting a list of said peer groups; receiving a user selection of one or more of said peer groups from said list; and outputting at least one of said pre-defined parameters for the or each selected peer groups.","267.A company board data processing method as in claim 266, wherein said pre-defined parameters relate to at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.","268.A company board data processing method as in claim 266, further comprising processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and/or one or more peer group parameter deviations; wherein any of said processed parameters are output.","269.A company board data processing method as in claim 266, further comprising storing parameters related to board members of a plurality of boards organised into boards; receiving a user selection of at least one said board for comparison with any said board peer group; and outputting the board member parameters for each member of said at least one board individually, to allow the or each board to be the subject of analysis.","270.A company board data processing method as in claim 266, further comprising receiving a user selection of one or more board members for comparison with any said board member peer group, and outputting the parameters of said board members, to allow the board members to be the subject of analysis.","271.A company board data processing method as in claim 266, further comprising receiving a user amendment of the constituents of said peer groups.","272.A company board data processing method as in claim 271, further comprising receiving an instruction from a user to copy one or more constituents of one said peer group to another said peer group.","273.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 266.274.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 266.275.A company board data processing system comprising: a board member store storing board member related parameters for a plurality of board members; parameter deviation determining means for determining deviations of said board member related parameters from pre-determined parameters; and a user interface generator for generating a user interface to display at least one of said board member related parameters highlighted in accordance with said determined deviations.","276.A company board data processing system as in claim 275, wherein said user interface generator is arranged to highlight each said board member related parameter in one of a plurality of colours, wherein said colours relate to the relative value of said determined deviations.","277.A company board data processing system as in claim 275, wherein said board member related parameters relate to one or more parameter totals, one or more parameter averages and/or one or more parameter deviations of a plurality of said board members.","278.A company board data processing system as in claim 275, wherein said predetermined parameters relate to a group of said board members, comprising a peer group.","279.A company board data processing system as in claim 278, wherein said predetermined parameters relate to at least one of one or more totals, one or more averages and one or more deviations of the parameters of the members of said peer group.","280.A company board data processing system as in claim 275, further comprising a parameter processor for processing parameters to determine at least one of one or more totals, one or more averages and one or more deviations for said pre-determined parameters; and wherein said user interface generator is adapted to generate said user interface to display said processed parameters.","281.A company board data processing system as in claim 275, wherein said parameter deviation determining means is adapted to determine said deviations as a number of standard deviations by which said board member related parameters differ from said pre-determined parameters.","282.A company board data processing system as in claim 275, wherein said user interface generator is adapted to generate a user interface to display a table of data, comprising a plurality of cells, each cell containing a said board member related parameter or a said pre-determined parameter, wherein said user interface generator is adapted to highlight said cells containing said board member related parameters in accordance with said determined deviation.","283.A company board data processing system as in claim 282, wherein said user interface generator is arranged to highlight each said cell in one of a plurality of colours, wherein said colours relate to the relative value of said determined deviations.","284.A company board data processing system as in claim 275, wherein said pre-determined parameters relate to a user selected and/or pre-defined group of board members arranged into a peer group.","285.A company board data processing system as in claim 284 further comprising: user selector means for receiving a user selection of at least one board and/or board member for inclusion in a peer group; and a parameter processor for processing parameters for constituents of a peer group to determine said predetermined parameters as at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.","286.A company board data processing system as in claim 285, wherein said user selector means is arranged to receive a user selection of one or more boards for comparison with at least one of said board related peer groups, and said user interface generator is adapted to generate the user interface to display the parameters related to members of said board highlighted in accordance with said determined deviation, to allow the board to be the subject of analysis.","287.A company board data processing system as in claim 285, wherein said user interface generator is adapted to generate a user interface to display the parameters related to individual members of the or each selected board highlighted in accordance with said determined deviation.","288.A company board data processing system as in claim 285, wherein said user selector means is arranged to receive a user selection of at least one board member for comparison with the board member peer group, and said user interface generator is adapted to generate the user interface to display the parameters related to said at least one board member highlighted in accordance with said determined deviations, to allow the or each board member to be the subject of analysis.","289.A company board data processing system as in claim 285, further comprising a peer group data store storing data for one or more user selected peer groups; wherein said user interface generator is adapted to generate a user interface to display a list of existing peer groups based on said stored peer group data; said user selector means is arranged to receive a user selection of one or more peer groups from said list; and said user interface generator is adapted to generate the user interface to display at least one of said processed parameters for at least one of said selected peer groups highlighted in accordance with said determined deviations.","290.A company board data processing system as in claim 285, further comprising a comparison group amendment means allowing a user to amend the constituents of a peer group.","291.A company board data processing system as in claim 290, wherein said peer group amendment means is adapted to allow a user to copy one or more constituents of a peer group to another peer group.","292.A company board data processing system as in claim 285, further comprising a searching means arranged to allow a user to search for a particular board in the board member store by inputting a specific company or board member name.","293.A company board data processing system as in claim 292, wherein said searching means is adapted to allow a user to search for a particular board in said board member store by selecting one or more parameters from a pre-defined list.","294.A company board data processing system as in claim 285, further comprising a store storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; wherein said user interface generator is adapted to generate a user interface to display a list of said pre-defined peer groups; including per group user selector means arranged to receive a user selection of one or more said pre-defined peer groups from said list; wherein said user interface generator is adapted to generate the user interface to display at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.","295.A company board data processing system as in claim 294, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.","296.A company board data processing method comprising: storing board member related parameters for a plurality of board members; determining deviations of said board member related parameters from pre-determined parameters; and generating a user interface to display at least one of said board member related parameters highlighted in accordance with said determined deviations.","297.A company board data processing method as in claim 296, wherein said user interface is generated to highlight each said board member related parameter in one of a plurality of colours, wherein said colours relate to the relative value of said determined deviations.","298.A company board data processing method as in claim 296, wherein said board member related parameters relate to one or more parameter totals, one or more parameter averages and/or one or more parameter deviations of a plurality of said board members.","299.A company board data processing method as in claim 296, wherein said pre-determined parameters relate to a group of said board members, comprising a peer group.","300.A company board data processing method as in claim 299, wherein said pre-determined parameters relate to at least one of one or more totals, one or more averages and one or more deviations of the parameters of the members of said peer group.","301.A company board data processing method as in claim 296, further comprising processing parameters to determine at least one of one or more totals, one or more averages and one or more deviations for said pre-determined parameters; and wherein said user interface displays said processed parameters.","302.A company board data processing method as in claim 296, wherein said determined deviations comprise a number of standard deviations by which said board member related parameters differ from said pre-determined parameters.","303.A company board data processing method as in claim 296, wherein said user interface displays a table of data comprising a plurality of cells, each cell containing a said board member related parameter or a said pre-determined parameter, said cells containing said board member related parameters being highlighted in accordance with said determined deviation.","304.A company board data processing method as in claim 303, wherein said user interface highlights each said cell in one of a plurality of colours, wherein said colours relate to the relative value of said determined deviations.","305.A company board data processing method as in claim 296 wherein said pre-determined parameters relate to a user selected and/or pre-defined group of board members arranged into a peer group.","306.A company board data processing method as in claim 305, further comprising: receiving a user selection of at least one board and/or board member for inclusion in a peer group; and processing parameters for constituents of a peer group to determine said predetermined parameters as at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.","307.A company board data processing method as in claim 306, wherein a user selection of one or more boards is received for comparison with at least one of said board related peer groups, and said user interface displays the parameters related to members of said board highlighted in accordance with said determined deviation, to allow the board to be the subject of analysis.","308.A company board data processing method as in claim 306, wherein said user interface displays the parameters related to individual members of the or each selected board highlighted in accordance with said determined deviation.","309.A company board data processing method as in claim 306, wherein a user selection of at least one board member is received for comparison with the board member peer group, and said user interface displays the parameters related to said at least one board member highlighted in accordance with said determined deviations, to allow the or each board member to be the subject of analysis.","310.A company board data processing method as in claim 306, further comprising storing data for one or more user selected peer groups in a peer group data store; wherein said user interface displays a list of existing peer groups based on said stored peer group data; a user selection of one or more peer groups from said list is received; and said user interface displays at least one of said processed parameters for at least one of said selected peer groups highlighted in accordance with said determined deviations.","311.A company board data processing method as in claim 306, further comprising receiving a user amendment of the constituents of a peer group.","312.A company board data processing method as in claim 311, wherein an instruction is received from a user to copy one or more constituents of a peer group to another peer group.","313.A company board data processing method as in claim 306, further comprising receiving a specific company or board member name from a user to search for a particular board in the board member store.","314.A company board data processing method as in claim 313, wherein a user selection of one or more parameters from a pre-defined list is received to search for a particular board in said board member store.","315.A company board data processing method as in claim 306, further comprising storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; wherein said user interface is generated to display a list of said pre-defined peer groups; a user selection of one or more said pre-defined peer groups from said list is received; and said user interface generates the user interface to display at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.","316.A company board data processing method as in claim 315, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.","317.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 296.318.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 296.319.A company board data processing system comprising: a board member store storing board member related parameters for a plurality of boards; and a user interface generator arranged to generate and display a multi axis plot of one or more of said board member related parameters for one or more boards and/or board members for comparison purposes.","320.A company board data processing system as in claim 319, further comprising a processor for determining a scale for at least one axis on said multi axis plot based on at least one of one or more totals, one or more averages and one or more deviations of a plurality of said parameters; wherein said user interface generator is adapted to display at least one axis on said multi axis plot with a scale equal to said determined scale.","321.A company board data processing system as in claim 319, wherein said board member store stores said parameters such that said board members are grouped into groups, and said user interface generator is arranged to generate a display of a multi axis plot of at least one of one or more totals, one or more averages or one or more deviations of said parameters of one or more said groups.","322.A company board data processing system as in claim 321, wherein said group comprises a board.","323.A company board data processing system as in claim 321, further comprising a parameter processor adapted to process parameters for said groups to determine at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations; wherein said user interface generator is adapted to display a plurality of said parameters and/or processed parameters on a multi axis plot, showing each axis for a different parameter type.","324.A company board data processing system as in claim 323, wherein said processor is adapted to determine a scale of at least one axis on said multi axis plot based on a predetermined level of deviation of at least one parameter already displayed on that axis from an average of a plurality of said stored and/or said processed parameters for a said group or a plurality of boards and/or board members.","325.A company board data processing system as in claim 323, wherein said parameter processor is adapted to determine the scale of at least one parameter axis by determining a minimum axis value of −n standard deviations and a maximum axis value of +m standard deviations of at least one parameter displayed on that axis from an average of a plurality of stored and/or processed parameters related to said group or a plurality of boards and/or board members, where n and m are integers.","326.A company board data processing system as in claim 325, wherein said plurality of boards and/or board members comprise all of said boards and/or board members from a specific pre-defined geographic region.","327.A company board data processing system as in claim 319 wherein said board member store stores parameters for at least some of said board members as a user selected and/or pre-defined group of board members assigned as a peer group.","328.A company board data processing system as in claim 327 further comprising: user selector means for receiving a user selection of at least one board and/or board member for inclusion in a peer group; a parameter processor for processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; and said user interface generator is adapted to generate the user interface to display a multi axis plot of one or more said processed parameters related to said peer group.","329.A company board data processing system as in claim 328, wherein said user selector means is arranged to receive a user selection of one or more boards for comparison with at least one of said board peer groups, and said user interface generator is adapted to generate a user interface to display a multi axis plot of one or more parameters related to members of said at least one board, to allow the or each board to be the subject of analysis.","330.A company board data processing system as in claim 328, wherein said user interface generator is adapted to generate a user interface to display a multi axis plot of the parameters related to individual members of the or each selected board.","331.A company board data processing system as in claim 328, wherein said user selector means is arranged to receive a user selection of at least one board member for comparison with said board member related peer group, and said user interface generator is adapted to generate a user interface to display a multi axis plot of the parameters related to said at least one board member, to allow the or each board member to be the subject of analysis.","332.A company board data processing system as in claim 328, wherein said user interface generator is adapted to generate a user interface to display a list of existing peer groups based on stored peer group data in said board member store; said user selector means is arranged to receive a user selection of one or more peer groups from said list; and said user interface generator is adapted to generate a user interface to display a multi axis plot of at least one of said processed parameters for at least one of said selected peer groups.","333.A company board data processing system as in claim 328, further comprising a comparison group amendment means allowing a user to amend the constituents of a peer group.","334.A company board data processing system as in claim 333, wherein said peer group amendment means is adapted to allow a user to copy one or more constituents of a peer group to another peer group.","335.A company board data processing system as in claim 328, further comprising a searching means arranged to allow a user to search for a particular board in the board member store by inputting a specific company or board member name.","336.A company board data processing system as in claim 335, wherein said searching means is adapted to allow a user to search for a particular board in said board member store by selecting one or more parameters from a pre-defined list.","337.A company board data processing system as in claim 328, wherein said user interface generator is adapted to generate a user interface to display a list of said pre-defined peer groups; said user selector means is arranged to receive a user selection of at least one said pre-defined peer groups from said list; and said user interface generator is adapted to generate a user interface to display a multi axis plot of at least one of said stored pre-defined peer group related parameters for said at least one selected pre-defined peer group.","338.A company board data processing system as in claim 337, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.","339.A company board data processing method comprising: storing board member related parameters for a plurality of boards; and generating a user interface displaying a multi axis plot of one or more of said board member related parameters for one or more boards and/or board members for comparison purposes.","340.A company board data processing method as in claim 339, further comprising determining a scale for at least one axis on said multi axis plot based on at least one of one or more totals, one or more averages and one or more deviations of a plurality of said parameters; wherein said user interface is generated to display at least one axis on said multi axis plot with a scale equal to said determined scale.","341.A company board data processing method as in claim 339, wherein said parameters are stored such that said board members are grouped into groups, and said user interface is generated to display a multi axis plot of at least one of one or more totals, one or more averages or one or more deviations of said parameters of one or more said groups.","342.A company board data processing method as in claim 341, wherein said group comprises a board.","343.A company board data processing method as in claim 341, further comprising processing parameters for said groups to determine at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations; wherein said user interface is generated to display a plurality of said parameters and/or processed parameters on a multi axis plot, showing each axis for a different parameter type.","344.A company board data processing method as in claim 343, wherein a scale of at least one axis on said multi axis plot is determined based on a predetermined level of deviation of at least one parameter already displayed on that axis from an average of a plurality of said stored and/or said processed parameters for a said group or a plurality of boards and/or board members.","345.A company board data processing method as in claim 343, wherein the scale of at least one parameter axis is determined by determining a minimum axis value of −n standard deviations and a maximum axis value of +m standard deviations of at least one parameter displayed on that axis from an average of a plurality of stored and/or processed parameters related to said group or a plurality of boards and/or board members, where n and m are integers.","346.A company board data processing method as in claim 345, wherein said plurality of boards and/or board members comprise all of said boards and/or board members from a specific pre-defined geographic region.","347.A company board data processing method as in claim 339 wherein said board member store stores parameters for at least some of said board members as a user selected and/or pre-defined group of board members assigned as a peer group.","348.A company board data processing method as in claim 347 further comprising: receiving a user selection of at least one board and/or board member for inclusion in a peer group; and processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; wherein said user interface generates the user interface to display a multi axis plot of one or more said processed parameters related to said peer group.","349.A company board data processing method as in claim 348, wherein a user selection of one or more boards is received for comparison with at least one of said board peer groups, and said user interface is generated to display a multi axis plot of one or more parameters related to members of said at least one board, to allow the or each board to be the subject of analysis.","350.A company board data processing method as in claim 348, wherein said user interface is generated to display a multi axis plot of the parameters related to individual members of the or each selected board.","351.A company board data processing method as in claim 348, wherein a user selection of at least one board member is received for comparison with said board member related peer group, and said user interface is generated to display a multi axis plot of the parameters related to said at least one board member, to allow the or each board member to be the subject of analysis.","352.A company board data processing method as in claim 348, wherein said user interface is generated to display a list of existing peer groups based on stored peer group data in said board member store; the method including receiving a user selection of one or more peer groups from said list; wherein said user interface is generated to display a multi axis plot of at least one of said processed parameters for at least one of said selected peer groups.","353.A company board data processing method as in claim 348, further comprising receiving a user amendment of the constituents of a peer group.","354.A company board data processing method as in claim 353, wherein an instruction is received from a user to copy one or more constituents of a peer group to another peer group.","355.A company board data processing method as in claim 348, further comprising receiving a specific company or board member name from a user to search for a particular board in the board member store.","356.A company board data processing method as in claim 355, wherein one or more parameters from a pre-defined list selected by a user is received to search for a particular board in said board member store.","357.A company board data processing method as in claim 348, wherein said user interface is generated to display a list of said pre-defined peer groups; a user selection of at least one said pre-defined peer groups from said list is received; and said user interface is generated to display a multi axis plot of at least one of said stored pre-defined peer group related parameters for said at least one selected pre-defined peer group.","358.A company board data processing method as in claim 357, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.","359.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 339.360.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 339.361.A company board data processing system comprising: access means for accessing a board member store storing parameters related to board members of a plurality of boards organised into boards; and a user interface generator for generating a user interface displaying said parameters related to board members for a plurality of boards to allow comparison of boards, at board member level, and/or board members of boards.","362.A company board data processing system as in claim 361, wherein said user interface generator is adapted to generate said user interface to display at least one of one or more totals, averages and deviations of said parameters related to a group of board members or boards to allow comparison with at least one board or board member.","363.A company board data processing method comprising: accessing a board member store storing parameters related to board members of a plurality of boards organized into boards; and generating a user interface displaying said parameters related to board members for a plurality of boards to allow comparison of boards, at board member level, and/or board members of boards.","364.A company board data processing method as in claim 363, wherein said user interface is generated to display at least one of one or more totals, averages and deviations of said parameters related to a group of board members or boards to allow comparison with at least one board or board member.","365.A company board data processing system comprising: a program memory storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 363.366.A carrier medium carrying computer readable instructions for controlling a computer to carry out the method of claim 363.367.A company board data processing system comprising: a store storing a number of shares owned by at least one board member in a company in which they are a board member; means to determine current share price of the company in which said board member is a member of the board; processor to process said number of shares and said share price to determine the value of said shares held by said board member; to determine a wealth index value for at least one said board member, comprising an increase in wealth related to said company of said board member when said share price increases by one percent; and output means for outputting at least one of said determined wealth index values.","368.A company board data processing method comprising: storing a number of shares owned by at least one board member in a company in which they are a board member; determining current share price of the company in which said board member is a member of the board; processing the number of shares and the share price to determine the value of said shares held by said board member; determining a wealth index for at least one said board member, comprising an increase in wealth related to said company of said board member when said share price increases by 1%; and outputting at least one of said determined wealth index values.","369.A company board data processing system, comprising: a store storing history information for a plurality of board members, said history information including information on previous companies of which said board member has been a board member, and timing information; a graphical display device for generating a graphical display showing performance data for companies together with the history data as a graph showing when the board member was at each company to allow a user to see how the companies have performed in relation to whether or not the board member was a member of the board for the company.","370.A system for analysing a created company board, comprising; a store for storing parameters related to a plurality of board members; user selection means for receiving a user selection of at least one board member for inclusion in at least one created company board; a parameter processor to generate board parameters for constituents of said created company board; and output means for outputting said board member parameters and/or said generated board parameters to allow the created board to be the subject of analysis.","371.A method of analyzing a created company board, comprising: storing parameters related to a plurality of board members; receiving a user selection of at least one board member for inclusion in at least one created company board; generating board parameters for constituents of said created company board; and outputting said company member parameters and/or said generated board parameters to allow the created board to be the subject of analysis.","372.A system for analyzing a created company board, comprising: a program memory storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 371.373.A carrier medium carrying computer readable instructions for controlling a computer to carry out the method of claim 371.374.A board member level recruitment and analysis tool comprising: a store storing parameters related to a plurality of board members; user selection means for receiving a user selection of at least one board member to be the subject of analysis, and a user selection of at least one other board member to be used as a comparison in board member analysis; and a display generator arranged to display the parameters of said at least one user selected board member subject to analysis and said at least one user selected comparison board member to allow recruitment related comparison of the two sets of parameters.","375.A board member level recruitment and analysis tool as in claim 374, further comprising: a processor for processing said board member parameters to determine at least one of one or more parameter totals, one or more parameter averages and one or more parameter deviations for a plurality of said user selected board members; wherein said display generator is adapted to display said processed parameters.","376.A board member level recruitment and analysis method comprising: storing parameters related to a plurality of board members; receiving a user selection of at least one board member to be the subject of analysis, and a user selection of at least one other board member to be used as a comparison in board member analysis; and displaying the parameters of said at least one user selected board member subject to analysis and said at least one user selected comparison board member to allow recruitment related comparison of the two sets of parameters.","377.A board member level recruitment and analysis system, comprising: a program memory storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to: store parameters related to a plurality of board members; receive a user selection of at least one board member to be the subject of analysis, and a user selection of at least one other board member to be used as a comparison in board member analysis; and displaying the parameters of said at least one user selected board member subject to analysis and said at least one user selected comparison board member to allow recruitment related comparison of the two sets of parameters.","378.A company board data processing system comprising: a board member store storing board member names for a plurality of board members organized into boards; association determining means for determining associations between said board members; a user interface generator for generating a user interface to display at least one of said board member names and a corresponding list of said determined associations for said board member.","379.A company board data processing system as in claim 378, wherein said association determining means is adapted to determine an association to exist between two board members when said two board members both serve on the same two or more independent boards together.","380.A company board data processing system as in claim 378, wherein said association determining means is adapted to determine an association to exist between two boards when one board member serves on said two boards at the same time.","381.A company board data processing system as in claim 378, wherein said association determining means is adapted to determine indirect associations between two or more boards and/or board members via a plurality of other associations; and said user interface generator is adapted to generate a user interface to display at least one board and/or board member and a list of said network of determined associations to at least one other board and/or board member.","382.A company board data processing system as in claim 378, wherein said user interface generator is adapted to generate a user interface to display a list of board members and/or boards based on said stored board member data; further comprising a user selector means for receiving a user selection of two or more boards and/or board members from said list; wherein said user interface generator is adapted to generate a user interface to display said names of said boards and/or board members and a list of said determined associations between said boards and/or board members.","383.A company board data processing system as in claim 381, wherein said user interface generator is adapted to generate a user interface to display a list of board members and/or boards based on said stored board member data; further comprising a user selector means for receiving a user selection of two or more boards and/or board members from said list; wherein said user interface generator is adapted to generate a user interface to display said names of said boards and/or board members and a list of said determined indirect associations between said boards and/or board members.","384.A company board data processing method comprising: storing board member names for a plurality of board members organized into boards; determining associations between said board members; generating a user interface to display at least one of said board member names and a corresponding list of said determined associations for said board member.","385.A company board data processing method as in claim 384, wherein said determining of associations includes determining an association to exist between two board members when said two board members both serve on the same two or more independent boards together.","386.A company board data processing method as in claim 384, wherein said determining of associations includes determining an association to exist between two boards when one board member serves on said two boards at the same time.","387.A company board data processing method as in claim 384 further comprising; determining indirect associations between two or more boards and/or board members via a plurality of other associations; wherein said user interface displays at least one board and/or board member and a list of said network of determined associations to at least one other board and/or board member.","388.A company board data processing method as in claim 384, wherein said user interface displays a list of board members and/or boards based on said stored board member data; further comprising receiving a user selection of two or more boards and/or board members from said list; and in response to said user selection said user interface displays said names of said selected boards and/or board members and a list of said determined associations between said boards and/or board members.","389.A company board data processing method as in claim 387, wherein said user interface displays a list of board members and/or boards based on said stored board member data; further comprising receiving a user selection of two or more boards and/or board members from said list; and in response to said user selection said user interface displays said names of said selected boards and/or board members and a list of said determined indirect associations between said boards and/or board members.","390.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 378.391.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 378.392.A method of fund management using the system of claim 1.393.A method of fund management using the system of claim 235.394.A method of fund management using the system of claim 259.395.A method of fund management using the system of claim 275.396.A method of fund management using the system of claim 319.397.A method of fund management using the system of claim 361.398.A method of fund management using the system of claim 367.399.A method of fund management using the system of claim 369.400.A method of fund management using the system of claim 370.401.A method of fund management using the system of claim 374.402.A method of fund management using the system of claim 375.403.A method of fund management using the system of claim 378.404.A method of recruiting board members using the system of claim 1.405.A method of recruiting board members using the system of claim 235.406.A method of recruiting board members using the system of claim 259.407.A method of recruiting board members using the system of claim 275.408.A method of recruiting board members using the system of claim 319.409.A method of recruiting board members using the system of claim 361.410.A method of recruiting board members using the system of claim 367.411.A method of recruiting board members using the system of claim 369.412.A method of recruiting board members using the system of claim 370.413.A method of recruiting board members using the system of claim 374.414.A method of recruiting board members using the system of claim 375.415.A method of recruiting board members using the system of claim 378."],["<SOH> BACKGROUND <EOH>Investing on the Stock Markets inevitably involves a risk on behalf of the investor, equities can go up or down and there is never any guarantee of a positive return from your investment.","However this uncertainty can be reduced considerably by investing wisely and more importantly by investing only in things that you have knowledge of.","You will have more chance of winning if you bet on a horse based on its form than you will if you take a guess from a list of starters without any knowledge of any of them, and so it is with the equity markets that a sensible investor will only buy or sell a stock based on sound information.","It is the requirement for this information across the stock markets of the world which has led to the existence of the world-wide Investment Banks and a plethora of financial advisors.","It is almost impossible for a fund manager or private investor to be able to make an informed decision on a stock without some form of advice on the economic conditions of that company, its past history, future prospects, products, staff, etc.",".",".",".","This advice can come in many forms depending on the information sought, for instance there are a large number of sources of company focussed economic data, information such as the price/earnings ratio, the profit & loss figures etc .",".",".",", well known financial statistics which can give an indication as to the stock's current worth and potential.","However, there are other forms of information that are more subjective by their very nature, and many of the large investment houses have teams of analysts trying to get behind the financial figures to tell the real story.","The problem with such information is that it is not as reliable as the hard economic facts, it often emanates from person to person meetings, press conferences and rumour.","It is not possible for a fund manager or investor to attend every annual general meeting, read every end of year report and interview every Board of Directors, and hence it is the analyst's job to try to do this for the investor, and it is left to the investor to attempt to read the analyst's report and pick out the information that he is looking for.","This, inevitably increases the risk of following the information and advice supplied, especially as two different analysts will often have a completely opposite view on the potential of any particular company.","Unfortunately there are very few alternatives, and whilst the use of analyst reports helps it still rarely allows for comparison across the whole spectrum of the market because analysts will often have a specialised sector and hence comparative data across a range of stocks in this field of information can be difficult to find and often lacking in user friendliness.","Hence it is the object of the proposed invention to supply a user friendly tool to aid in information provision."],["<SOH> SUMMARY OF THE INVENTION <EOH>A first aspect of the present invention provides a company board data processing system and method which provides a user interface platform to supply information related to members of boards of companies to enable comparison of board members or boards at board member level.","Parameters related to board members of a plurality of boards are stored.","The boards of which the board members are members is indicated in the stored parameters.","A user interface is generated which comprises a display of the parameters related to board members for a plurality of boards and/or individual board members to, allow comparison of board members and/or boards at board member level.","Thus this aspect of the present invention provides a tool which can be used by analysts to review parameters related to boards at the level of granularity of board member parameters.","In one embodiment of this aspect of the present invention, parameters for a group of board members such as a board or some other predetermined group is processed to determine process parameters which can comprise one or a number of: parameter totals, parameter averages and parameter deviations.","Thus the user interface can display such process parameters for comparison with boards at board member level or with individual board members.","In one embodiment of this aspect of the present invention the group can comprise a group of boards in which case the process parameters comprise parameters processed for a number of boards and thus represents totals, averages or deviations for the boards.","This enables the board as a whole and individual members of the board to be compared to a number of boards.","In an alternative embodiment of this aspect of the present invention the group comprises a group of board members.","The group of board members can comprise board members of a single board, or board members across a number of boards.","This enables individual board members to be compared with a peer group of board members from a number of boards.","A second aspect of the present invention provides a company board data processing system and method in which parameters related to board members of a plurality of boards is stored.","The system receives a user selection of at least one board and/or board member for inclusion in a peer group.","Parameters for constituents of a peer group are processed to determine processed parameters for the peer group and these processed parameters are output.","Preferably the processing determines one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.","This aspect of the present invention provides a useful tool to enable a user to select and form a peer group comprised of a number of board members which need not be in the same board.","This aspect of the present invention also allows the selection of boards for inclusion in a board peer group.","Parameters for boards are processed to determine process parameters for the board peer group.","Thus a board peer group can be used for comparison with a board to allow comparative analysis.","In one embodiment of this aspect of the present invention the peer group data is stored for at least one user selected peer group.","The user interface can display a list of existing peer groups to allow a user to select one or more of the existing peer groups.","Process parameters for the or each selected peer group can then be output for display, e.g.","for comparative purposes.","In one embodiment of this aspect of the present invention, the constituents of the peer groups can be amended.","This amendment can take the form of allowing constituents of a peer group, e.g.","a board member or board, to be transferred from one peer group to another.","In another embodiment of this aspect of the present invention predefined groups of boards and/or board members are stored with parameters related to the peer groups as predefined peer groups.","The user interface can display a list of the predefined peer groups to allow a user to select one or more of them for output for comparative purposes.","Another aspect of the present invention provides a company board data processing system in which predefined groups of boards and/or board members comprising peer groups and predefined parameters related to the peer groups are stored.","The system is arranged to output a list of the peer groups to allow a user to select one or more of the peer groups from the list.","As a result of the selection at least one of the predefined parameters for the or each selected peer group is output.","The parameters can be displayed and used for comparative purposes.","In this aspect of the present invention a peer group which can comprise parameters for groups of boards and/or board members is predefined.","In one embodiment, however, a user can modify a predefined peer group by adding constituents, i.e.","boards or board members, or removing constituents.","Also, constituents can be moved from group to group.","Thus this embodiment provides a flexible tool for comparison of board members and boards with peer groups.","In one embodiment the predefined parameters for the peer groups comprise process parameters.","The process parameters can comprise peer group parameter totals, peer group parameter averages and peer group parameter deviations.","Another aspect of the present invention provides a company board data processing system and method in which board member related parameters for a plurality of board members is stored.","A deviation of said board member related parameters from predetermined parameters is determined and this is used by a user interface to generate a display of at least one board member related parameter highlighted in accordance with the determined deviations.","This embodiment of the present invention provides a system in which deviations of parameters related to board members from predetermined parameters is displayed in a clear form by highlighting such parameters in accordance with the deviations.","This allows these deviations and patterns in these deviations to be easily distinguishable.","In one embodiment of the present invention the user interface is generated to highlight each board member related parameter in one of a plurality of colours, where the colour relates to the relative value of the determined deviation.","In one embodiment of this aspect of the present invention the predetermined parameters relate to a group of board members comprising a peer group.","In this embodiment predetermined parameters can relate to totals, averages and/or deviations of the parameters of the members of the peer group.","In one embodiment of this aspect of the present invention the user interface generates a table of data comprising a plurality of cells.","Each cell contains a board member related parameter or a predetermined parameter.","Cells containing the board member related parameters are highlighted in accordance with the determined deviation.","In a preferred embodiment these cells are highlighted in one of a plurality of colours, wherein the colours relate to the relative value of the determined deviations.","In another aspect of this aspect of the present invention there is provided a company board data processing system and method in which board member related parameters are stored for a plurality of boards.","A user interface generates a multi-axis plot of one or more of the board member related parameters for one or more boards and/or board members to allow comparison of these parameters.","Thus this aspect of the present invention provides a graphical user interface tool for the visualization of company board data at a company board member level of granularity.","In one embodiment of this aspect of the present invention the board members can be grouped into groups and the user interface can generate a display of a multi-axis plot of parameters related to a group of board members.","In one embodiment this can be used for comparative purposes to allow parameters related to a board or individual members of a board to be plotted against the group.","In one embodiment of the present invention the axes of the multi-axis plot can be scaled based on a plurality of parameters for a plurality of board members.","This can comprise totals, averages or deviations for parameters for a plurality of board members.","A further aspect of the present invention provides a company board data processing system in which a number of shares owned by at least one board member in a company in which they are a board member are stored.","The current share price of the company in which the board member is a member of the board is determined and used with the number of shares to determine the value of the shares held by the board member to determine a wealth index for the board member.","The wealth index comprises an increase in wealth related to the company when the share price increases by an amount, e.g.","1%.","This aspect of the present invention provides an indicator of the wealth related motivation of a board member to improve the performance of a company.","If a board member holds a large number of shares, they have a personal interest in improving the performance of a company so that the share price increases.","Another aspect of the present invention provides a system for analyzing a created company board.","Parameters related to a plurality of board members are stored.","A user can select at least one board member for inclusion in at least one created company board.","Board parameters for constituents of the created company board are generated.","The board member parameters and/or the generated board parameters can be output to allow the created board to be the subject of analysis.","Thus this aspect of the present invention provides a means by which a user of the system can create a “fantasy” board comprised of a number of board members currently members of other boards.","Parameters related to the board members for the created company board can then be analysed.","Thus this aspect of the present invention can be used as a tool by which users can identify suitable board members for building a company board for a new company or for replacing board members of an existing company board.","A further aspect of the present invention provides a board member level recruitment and analysis tool and method in which parameters related to a plurality of board members are stored.","A user is able to select at least one board member to be the subject of analysis and to select at least one other board member to be used as a comparison in the board member analysis.","A user interface generates a display of the parameters of the selected board members subject to analysis and the selected comparison board members to allow recruitment related comparison of the two sets of parameters.","Another aspect of the present invention provides a company board data processing system and method in which a graphical display is generated to show the performance of a company and a board member history on a time basis.","This aspect of the present invention allows a user to identify the effect of a board member on the performance of a company.","Although the present invention has been described hereinabove with reference to a number of separate aspects, in accordance with the present invention, any aspect of the present invention described hereinabove can be used in conjunction with any other aspect of the present invention.","The present invention is applicable to financial analysis and in management of companies.","For example, the present invention can be used by fund managers for managing a portfolio of investments by analyzing company board member parameters.","The presentation of the data can be by any suitable output means such as a display or printer.","The output can also be an output over a network to provide for presentation, e.g.","display, at a remote location.","The present invention is applicable to a stand alone system or a network system in which the server receives requests from a user of a client and outputs data for presentation to the user by the client.","All aspects of the present invention can be implemented on dedicated hardware, a combination of dedicated hardware and suitably programmed hardware, or on a programmed general purpose computer.","The present invention thus encompasses computer readable instructions for controlling a computer to carry out the methods.","Computer readable instructions can be provided to a general purpose computer on any suitable carrier medium such as a transient medium, e.g.","an electrical, optical, microwave, acoustic or radio frequency signal (e.g.","a signal (carrier wave) carrying computer readable instructions over a network such as the Internet), or a storage medium such as a floppy disk, hard disk, CD-ROM or solid state memory device."],["FIELD OF INVENTION This invention relates to a data processing system and method for processing and presenting a wide range of data related to the Board of Directors of companies, as well as the members of those Boards individually.","BACKGROUND Investing on the Stock Markets inevitably involves a risk on behalf of the investor, equities can go up or down and there is never any guarantee of a positive return from your investment.","However this uncertainty can be reduced considerably by investing wisely and more importantly by investing only in things that you have knowledge of.","You will have more chance of winning if you bet on a horse based on its form than you will if you take a guess from a list of starters without any knowledge of any of them, and so it is with the equity markets that a sensible investor will only buy or sell a stock based on sound information.","It is the requirement for this information across the stock markets of the world which has led to the existence of the world-wide Investment Banks and a plethora of financial advisors.","It is almost impossible for a fund manager or private investor to be able to make an informed decision on a stock without some form of advice on the economic conditions of that company, its past history, future prospects, products, staff, etc.",".",".",".","This advice can come in many forms depending on the information sought, for instance there are a large number of sources of company focussed economic data, information such as the price/earnings ratio, the profit & loss figures etc .",".",".",", well known financial statistics which can give an indication as to the stock's current worth and potential.","However, there are other forms of information that are more subjective by their very nature, and many of the large investment houses have teams of analysts trying to get behind the financial figures to tell the real story.","The problem with such information is that it is not as reliable as the hard economic facts, it often emanates from person to person meetings, press conferences and rumour.","It is not possible for a fund manager or investor to attend every annual general meeting, read every end of year report and interview every Board of Directors, and hence it is the analyst's job to try to do this for the investor, and it is left to the investor to attempt to read the analyst's report and pick out the information that he is looking for.","This, inevitably increases the risk of following the information and advice supplied, especially as two different analysts will often have a completely opposite view on the potential of any particular company.","Unfortunately there are very few alternatives, and whilst the use of analyst reports helps it still rarely allows for comparison across the whole spectrum of the market because analysts will often have a specialised sector and hence comparative data across a range of stocks in this field of information can be difficult to find and often lacking in user friendliness.","Hence it is the object of the proposed invention to supply a user friendly tool to aid in information provision.","SUMMARY OF THE INVENTION A first aspect of the present invention provides a company board data processing system and method which provides a user interface platform to supply information related to members of boards of companies to enable comparison of board members or boards at board member level.","Parameters related to board members of a plurality of boards are stored.","The boards of which the board members are members is indicated in the stored parameters.","A user interface is generated which comprises a display of the parameters related to board members for a plurality of boards and/or individual board members to, allow comparison of board members and/or boards at board member level.","Thus this aspect of the present invention provides a tool which can be used by analysts to review parameters related to boards at the level of granularity of board member parameters.","In one embodiment of this aspect of the present invention, parameters for a group of board members such as a board or some other predetermined group is processed to determine process parameters which can comprise one or a number of: parameter totals, parameter averages and parameter deviations.","Thus the user interface can display such process parameters for comparison with boards at board member level or with individual board members.","In one embodiment of this aspect of the present invention the group can comprise a group of boards in which case the process parameters comprise parameters processed for a number of boards and thus represents totals, averages or deviations for the boards.","This enables the board as a whole and individual members of the board to be compared to a number of boards.","In an alternative embodiment of this aspect of the present invention the group comprises a group of board members.","The group of board members can comprise board members of a single board, or board members across a number of boards.","This enables individual board members to be compared with a peer group of board members from a number of boards.","A second aspect of the present invention provides a company board data processing system and method in which parameters related to board members of a plurality of boards is stored.","The system receives a user selection of at least one board and/or board member for inclusion in a peer group.","Parameters for constituents of a peer group are processed to determine processed parameters for the peer group and these processed parameters are output.","Preferably the processing determines one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.","This aspect of the present invention provides a useful tool to enable a user to select and form a peer group comprised of a number of board members which need not be in the same board.","This aspect of the present invention also allows the selection of boards for inclusion in a board peer group.","Parameters for boards are processed to determine process parameters for the board peer group.","Thus a board peer group can be used for comparison with a board to allow comparative analysis.","In one embodiment of this aspect of the present invention the peer group data is stored for at least one user selected peer group.","The user interface can display a list of existing peer groups to allow a user to select one or more of the existing peer groups.","Process parameters for the or each selected peer group can then be output for display, e.g.","for comparative purposes.","In one embodiment of this aspect of the present invention, the constituents of the peer groups can be amended.","This amendment can take the form of allowing constituents of a peer group, e.g.","a board member or board, to be transferred from one peer group to another.","In another embodiment of this aspect of the present invention predefined groups of boards and/or board members are stored with parameters related to the peer groups as predefined peer groups.","The user interface can display a list of the predefined peer groups to allow a user to select one or more of them for output for comparative purposes.","Another aspect of the present invention provides a company board data processing system in which predefined groups of boards and/or board members comprising peer groups and predefined parameters related to the peer groups are stored.","The system is arranged to output a list of the peer groups to allow a user to select one or more of the peer groups from the list.","As a result of the selection at least one of the predefined parameters for the or each selected peer group is output.","The parameters can be displayed and used for comparative purposes.","In this aspect of the present invention a peer group which can comprise parameters for groups of boards and/or board members is predefined.","In one embodiment, however, a user can modify a predefined peer group by adding constituents, i.e.","boards or board members, or removing constituents.","Also, constituents can be moved from group to group.","Thus this embodiment provides a flexible tool for comparison of board members and boards with peer groups.","In one embodiment the predefined parameters for the peer groups comprise process parameters.","The process parameters can comprise peer group parameter totals, peer group parameter averages and peer group parameter deviations.","Another aspect of the present invention provides a company board data processing system and method in which board member related parameters for a plurality of board members is stored.","A deviation of said board member related parameters from predetermined parameters is determined and this is used by a user interface to generate a display of at least one board member related parameter highlighted in accordance with the determined deviations.","This embodiment of the present invention provides a system in which deviations of parameters related to board members from predetermined parameters is displayed in a clear form by highlighting such parameters in accordance with the deviations.","This allows these deviations and patterns in these deviations to be easily distinguishable.","In one embodiment of the present invention the user interface is generated to highlight each board member related parameter in one of a plurality of colours, where the colour relates to the relative value of the determined deviation.","In one embodiment of this aspect of the present invention the predetermined parameters relate to a group of board members comprising a peer group.","In this embodiment predetermined parameters can relate to totals, averages and/or deviations of the parameters of the members of the peer group.","In one embodiment of this aspect of the present invention the user interface generates a table of data comprising a plurality of cells.","Each cell contains a board member related parameter or a predetermined parameter.","Cells containing the board member related parameters are highlighted in accordance with the determined deviation.","In a preferred embodiment these cells are highlighted in one of a plurality of colours, wherein the colours relate to the relative value of the determined deviations.","In another aspect of this aspect of the present invention there is provided a company board data processing system and method in which board member related parameters are stored for a plurality of boards.","A user interface generates a multi-axis plot of one or more of the board member related parameters for one or more boards and/or board members to allow comparison of these parameters.","Thus this aspect of the present invention provides a graphical user interface tool for the visualization of company board data at a company board member level of granularity.","In one embodiment of this aspect of the present invention the board members can be grouped into groups and the user interface can generate a display of a multi-axis plot of parameters related to a group of board members.","In one embodiment this can be used for comparative purposes to allow parameters related to a board or individual members of a board to be plotted against the group.","In one embodiment of the present invention the axes of the multi-axis plot can be scaled based on a plurality of parameters for a plurality of board members.","This can comprise totals, averages or deviations for parameters for a plurality of board members.","A further aspect of the present invention provides a company board data processing system in which a number of shares owned by at least one board member in a company in which they are a board member are stored.","The current share price of the company in which the board member is a member of the board is determined and used with the number of shares to determine the value of the shares held by the board member to determine a wealth index for the board member.","The wealth index comprises an increase in wealth related to the company when the share price increases by an amount, e.g.","1%.","This aspect of the present invention provides an indicator of the wealth related motivation of a board member to improve the performance of a company.","If a board member holds a large number of shares, they have a personal interest in improving the performance of a company so that the share price increases.","Another aspect of the present invention provides a system for analyzing a created company board.","Parameters related to a plurality of board members are stored.","A user can select at least one board member for inclusion in at least one created company board.","Board parameters for constituents of the created company board are generated.","The board member parameters and/or the generated board parameters can be output to allow the created board to be the subject of analysis.","Thus this aspect of the present invention provides a means by which a user of the system can create a “fantasy” board comprised of a number of board members currently members of other boards.","Parameters related to the board members for the created company board can then be analysed.","Thus this aspect of the present invention can be used as a tool by which users can identify suitable board members for building a company board for a new company or for replacing board members of an existing company board.","A further aspect of the present invention provides a board member level recruitment and analysis tool and method in which parameters related to a plurality of board members are stored.","A user is able to select at least one board member to be the subject of analysis and to select at least one other board member to be used as a comparison in the board member analysis.","A user interface generates a display of the parameters of the selected board members subject to analysis and the selected comparison board members to allow recruitment related comparison of the two sets of parameters.","Another aspect of the present invention provides a company board data processing system and method in which a graphical display is generated to show the performance of a company and a board member history on a time basis.","This aspect of the present invention allows a user to identify the effect of a board member on the performance of a company.","Although the present invention has been described hereinabove with reference to a number of separate aspects, in accordance with the present invention, any aspect of the present invention described hereinabove can be used in conjunction with any other aspect of the present invention.","The present invention is applicable to financial analysis and in management of companies.","For example, the present invention can be used by fund managers for managing a portfolio of investments by analyzing company board member parameters.","The presentation of the data can be by any suitable output means such as a display or printer.","The output can also be an output over a network to provide for presentation, e.g.","display, at a remote location.","The present invention is applicable to a stand alone system or a network system in which the server receives requests from a user of a client and outputs data for presentation to the user by the client.","All aspects of the present invention can be implemented on dedicated hardware, a combination of dedicated hardware and suitably programmed hardware, or on a programmed general purpose computer.","The present invention thus encompasses computer readable instructions for controlling a computer to carry out the methods.","Computer readable instructions can be provided to a general purpose computer on any suitable carrier medium such as a transient medium, e.g.","an electrical, optical, microwave, acoustic or radio frequency signal (e.g.","a signal (carrier wave) carrying computer readable instructions over a network such as the Internet), or a storage medium such as a floppy disk, hard disk, CD-ROM or solid state memory device.","BRIEF DESCRIPTION OF THE DRAWINGS Embodiments of the present invention will now be described with reference to the accompanying drawings, in which: FIG.","1 is a schematic diagram of a company board data processing system in accordance with an embodiment of the present invention; FIG.","2 illustrates the top level structure of the user interface information pages; FIG.","3 illustrates the information page structure of the Board Associations data section; FIG.","4 illustrates the information page structure of the Board Remuneration data section; FIG.","5 illustrates the information page structure of the Board Characteristics data section; FIG.","6 illustrates the information page structure of the Director Associations data section; FIG.","7 illustrates the information page structure of the Director Remuneration data section; FIG.","8 illustrates the information page structure of the Director Characteristics data section; FIG.","9 is a screen shot of a user interface ‘Board Summary’ information page; FIG.","10 is a screen shot of a user interface ‘Board Single & Multi Associations’ information page; FIG.","11 is a screen shot of a user interface ‘Board Remuneration Summary Table’ information page; FIG.","12 is a screen shot of a user interface ‘Board Remuneration Summary Bubble Chart’ information page; FIG.","13 is a screen shot of a user interface ‘Board Remuneration Ratios’ information page; FIG.","14 is a screen shot of a user interface ‘Board Characteristics Network/Biographical’ information page; FIG.","15 is a screen shot of a user interface ‘Board Characteristics Change Table’ information page; FIG.","16 is a screen shot of a user interface ‘Board Characteristics Diversity Table’ information page; FIG.","17 is a screen shot of a user interface ‘Board Characteristics Diversity Radar Chart information page; FIG.","18 is a screen shot of a user interface ‘Board Peer Group Management’ information page; FIG.","19 is a screen shot of a user interface ‘Board Peer Group Member’ information page; FIG.","20 is a screen shot of a user interface ‘Board Advanced Search’ information page, FIG.","21 is a screen shot of a user interface ‘Board Search Results’ information page; FIG.","22 is a screen shot of a user interface ‘Director Summary’ information page; FIG.","23 is a screen shot of a user interface ‘Director Single & Multi Associations’ information page; FIG.","24 is a screen shot of a user interface ‘Director Remuneration Ratios’ information page; FIG.","25 is a screen shot of a user interface ‘Director Characteristics Roles’ information page; FIG.","26 is a screen shot of a user interface ‘Director Characteristics Experience Table’ information page; FIG.","27 is a screen shot of a user interface ‘Director Characteristics Experience Radar Chart’ information page; FIG.","28 is a screen shot of a user interface ‘Director Peer Group Management’ information page; FIG.","29 is a screen shot of a user interface ‘Director Peer Group Member’ information page; FIG.","30 is a screen shot of a user interface ‘Director Advanced Search’ information page; FIG.","31 is a screen shot of a user interface ‘Director Search Results’ information page; FIG.","32 is a screen shot of a user interface ‘Board Timeline’ information page; FIG.","33 is a screen shot of a user interface ‘Board Share Price History’ information page; FIG.","34 is a screen shot of a user interface ‘Board and Shareholder Returns’ information page; FIG.","35 is a screen shot of a user interface ‘Board Time Analysis’ information page; FIG.","36 is a screen shot of a user interface ‘Board Experience over Time’ information page; FIG.","37 is a screen shot of a user interface ‘Director Multi Associations’ information page; FIG.","38 is a screen shot of a user interface ‘Director Share Price History’ information page; FIG.","39 is a screen shot of a user interface ‘Board Peer Group Alerts Set-Up’ information page; FIG.","40 is a screen shot of a user interface ‘Board Peer Group Alerts Results’ information page; FIG.","41 is a screen shot of a user interface ‘Director Peer Group Alerts Set-Up’ information page; and FIG.","42 is a screen shot of a user interface ‘Director Peer Group Alerts Results’ information page.","DESCRIPTION OF PREFERRED EMBODIMENTS General Introduction There are many systems that provide information regarding companies listed on the stock markets of the world, but that there is a gap in the market for an analysis tool that is focussed on the actual members of the Boards of these companies.","Such information is currently hard to find and more importantly difficult to analyse and often unreliable.","The system of the embodiment of the present invention allows people to view and analyse the people who actually run these listed companies and hence offer a platform for people to form an indicative view of another very important aspect in measuring the potential success, or otherwise, of such companies that has so far been very difficult to grasp.","The system offered by the embodiment of the current invention allows the user to select an entire Board and analyse and compare the credentials of each member of that Board.","Also, one or more other Boards can be selected, by the user, and corresponding data can be displayed simultaneously to allow a comparison of the selected Boards against one another, hence providing the statistics that can be used to measure the relative merits of one Board against another.","This system provides a platform of user interactive analysis that has not been previously available and that will greatly benefit anyone investing in stock market listed companies.","However, the usefulness of such a tool is not restricted to the financial markets.","Anyone who wished to compare the skills, experience and background history of one or more Director level individuals, say for recruitment purposes, would also find this tool very effective in their analysis.","The system allows the user to select an individual and view data on all of his Board-level roles, and also offers the chance to select one or more other individuals to be displayed simultaneously and used in the analysis.","Hardware The preferred embodiment of the current invention is illustrated in FIG.","1 and comprises an architecture structure that is made up of a Data Repository System, 1, a Database System, 2, 3, 4, 5, a Data Processor, 6, a web Server system, 7, 8, 9, and a User's computer, 11.The Data Repository System, 1, is located at a central data entry and management location, 100.It comprises a unified data entry point and is the tool with which all raw data is entered into the system.","The Data Repository System 1 acts as a master database for the data in the system and receives data via one of two methods: manual data entry; or automatic data entry.","The manual data entry system comprises a computer system with a user interface based at the central data entry and management location, 100.This user interface allows manual entry of new data raw data parameters as well as the entry of some manual analytics.","It comprises two interfaces, one for the entry of data relating to a Director Profile and the other for the entry of data relating to a Board profile.","The raw data comprises simple numerical values, such as a Director's salary, that are described hereafter.","The manual analytics data entry comprises such data as, for example, a company time-line share price graph that has been manually amended to display key corporate actions during the time period displayed, for example the departure of a CEO from the Board or a new company share issue, such as that shown in FIG.","33.The automatic data entry system comprises on on-line feed to a number of financial data providers, such as FT, AFX and Hernscott.","This data is downloaded periodically, daily in the case of the preferred embodiment of the invention, into a data store, where it is checked both by manual and automatic means before being populated onto the Master Database.","The raw data parameters that are entered via the manual and automatic data entry systems comprise parameters related to Board members and parameters related to Boards and include: Salary Bonus Pension Contributions Other remuneration Role Number of Options awarded Stock Price Option Exercise Price Options time to expiry Dividend Yield Number of Directors at the start of the Year Number of Directors at the end of the Year Equity awarded LTIP Contributions awarded Number of Executives and Non-executives in the company Age of Director Time in Company of Director Time on Board of Director Board's policy retirement Age Time to retirement for Director Name Role Director's Direct Compensation Director's sex Director's Nationality Director's past employment Director's Qualifications Director's Shared external Directorships Total Company Revenue EBITDA—earnings before interest tax depreciation amortisation Operating Profit Company's net income Company's fixed assets Company's current liabilities Company's long-term liabilities Company's long-term creditors Company's total assets Company's fixed assets Company's current assets Company's total liabilities Company's PE ratio Company's Auditors Company's Financial Advisors Company's Stock Brokers Company's Financial PR Company's Solicitors The central data entry and management location, 100, containing the Data Repository System, 1, acting as the Master Database is connected via a dedicated link to a slave Database system, 200, located at a different location.","The Database system, 200, provides the publicly accessible data as a mirror of the data in the Master Database.","The Database system, 200, comprises two systems, 300 and 500, each comprising a Database Processor, 4 and 5, connected to a Database, 2 and 3, which act as slaves to the Master Database.","There are two database systems to provide redundancy in the case of the failure of one system.","The data entered onto the system by means of the Data Repository System, 1, is loaded onto the two database processors, 4 and 5, which perform a series of analytical calculations to transform the raw data into a form that is useful for the system, e.g.","the calculation of complex variables, or data set averages and deviations.","The two databases, 2 and 3, are then populated with the plurality of predetermined calculated parameters, and the raw data, for storage.","In this way the Master Database is kept offline and this mirroring of data takes place regularly, e.g.","daily at night when usage is low.","This system enables the Master Database to be continuously updated at the central data entry and management location.","The two database processors, 4 and 5, are further connected to a data processor 6.The Data Processor 6 is used to perform user end requested calculations, for instance it will calculate peer group parameters, which are discussed hereafter.","These parameters are stored in the databases, 2 and 3, and are specific to users.","Thus user specific parameters such as Peer Group data are generated and stored in the databases, 2 and 3.The Database Processors, 4 and 5, download the data from the Databases, 2 and 3, and transmit the data to the Data Processor, 6.The Data Processor 6 is connected to two a third system, 500, that comprises two Web Servers, 7 and 8 and a Web Page Data Processor.","The system, 500, provides the method of accessing the data in the system, 200, i.e.","a web interface.","The connection is made across a firewall system for security.","Again the existence of two Web Servers is based on the necessity for redundancy in the system.","The Data Processor 6, acting as an Application Server to provide data to the Web Servers, 7 and 8, sorts and processes the data into a useable form for the user end of the system, and communicates that data onto the Web Servers, 7 and 8.The Web Servers, 7 and 8, are both connected to a Web Page Data Store, 9.By communicating with the Web Page Data Store, 9, the Web Servers, 7 and 8, generate web pages that are to be communicated to the client's user interface, based on the data received from the Data Processor, 6.The Web Servers, 7 and 8, are connected to an IP network, 10, e.g.","the internet.","A User's Web Browser, 12, on a User's Computer, 11, is also connected to said IP Network, 10.When requested by the User's Web Browser, 12, the Web Servers, 7 and 8, will transmit said generated web pages to the User's Web Browser, 12, via the IP Network, 10.Also the Web Servers, 7 and 8, can transmit Java Script and Java Applets to control the client's user interface, and to generate a number of the web pages on the User's Web Browser, 12.Thus the user interface can be server side generated, client side generated or a mixture of the two.","The User's Web Browser, 12, will then display said web pages in the form of a user interactive display on the User's Computer, 11.The User's Web Browser, 12, can also communicate requests for information back up through the system.","This may involve a request for new processed parameters, for example based on a new user selected peer group that the user has set up on his browser.","In this scenario a request would be transmitted back to the Data Processor, 6, via the IP network, 10, and one of the Web Servers, 7 or 8.The Data Processor will process the request and transmit the new processed parameters back to the User's Computer in the same manner as the initial parameters were transmitted as discussed above.","In this way the Data Processor, 6, generates user specific parameters, e.g.","peer groups, and these are stored, along with a User ID, in the databases, 2 and 3, to provide for a level of personalisation when a user accesses the data in the Databases, 2 and 3.User Interface Description In the embodiment of the present invention, the user interface displayed on the user's web browser is split into two main sections, as shown in FIG.","2: one supplies information regarding one or more Boards and offers comparative analysis of these Boards against other Boards; and the other offers the same functionality at Director level, allowing the comparison of one or more individual Directors against other Directors.","In both cases the system is able to display a large number of parameters for any Board and/or Director through a variety of medium, including tabular form and a number of graphical displays.","Data Fields Described by Screen FIG.","2 shows the main structure of the system's visual user interface information pages.","The pages are organised into a hierarchical structure, with each branch containing further levels of detail below it.","As mentioned, above, the user must first select either the Board related section or the Director related section.","This structure can clearly be seen in FIG.","2 where the Board related pages are on the left hand branch and the Director related pages are on the right hand branch.","In each of the two sections the user can select from a list of main topics headings: Summary Associations Remuneration Characteristics Profile Each of these options will display top-level data related to their topic.","Some of these topics have further layers of increasingly detailed data, and/or different graphical display methods, in a plurality of related screens and each of the data layers will now be discussed in more detail, initially by looking at the Board related branch and then the Director related branch.","Board Summary The Board Summary, as shown in FIG.","9, has two themes: firstly it provides a general overview of the members of a Board and displays the following basic information relating to those members: Executive Name- names of all of the Board Directors Role- The position held by the Director in the Company.","Time on Board- The length of time the Director has been on the Board (in years).","Time in Company- The length of time spent by the Director in the company (in years).","Time to Retirement- The length of time to retirement, based on the company's policy retirement age (in years).","Stakeholder Representative- specific stakeholder group(s) represented by the Director for example shareholders representatives, union, employees..","The second theme to the Board summary is a list of major shareholders in the company, giving their name and an indication as to the size of their holding.","Board Associations The Board Associations section has two levels beneath it, as shown in FIG.","3, named ‘Single and Multi Associations’, as shown in FIG.","10, and ‘Multi Associations’.","These two pages provide lists of associations that the Directors of a Board have with the Boards of other companies.","A ‘Single Association’ occurs when a Director on the Board of one particular company is also serving on the Board of another company, and is the only person from the subject company to have any association with that latter company.","A ‘Multi Association’ occurs when two or more Directors from one particular Board both have associations with another particular Board, i.e.","two or more Directors are currently serving on that other Board together, or at least one Director has served on another Board at the same time as at least another Director was also serving on that Board, if at least one of them is currently still sitting on that other board.","These two features can be seen on FIG.","10, where an example ‘Single Association’ is Georgh Ehrnrooth (bottom row), who is a Director on the Board of Nokia OJY, the subject company, and is also currently a Director on the Board of Wartsila OYJ ABP, and an example of a ‘Multi Association’ is Georg Ehrnrooth and Olli-Pekka Kallasvuo, fifth row from the bottom, who are both on the Board of the Subject company Nokia OYJ and are also both currently on the Board of Sampo OYJ.","Board Remuneration The Board Remuneration section can be broken down into five main sections: Remuneration Summary Direct Remuneration—annual cash based pay.","Equity Linked Remuneration—annual equity linked pay.","Equity Linked Wealth—total equity linked benefits received.","Remuneration Ratios This hierarchy can be seen in FIG.","4.These sections will now be discussed in turn.","The Remuneration Summary, as shown in FIG.","4, consists of four separate pages, each displaying the summary information in a different manner.","The Remuneration Summary Table, as shown in FIG.","11, displays the following parameters: Executive Name- names of all of the Board Directors Role- The position held by the Director in the Company.","Total Direct Remuneration- the sum of salary, bonus, pension (defined contribution) and other non-equity remuneration.","Total Equity Linked Remuneration- amount of remuneration in the form of equity, shares, and options for a particular financial period.","Total Remuneration for the Period- the sum of the Total Direct Remuneration and the Total Equity Linked Remuneration, each as described above.","Total Equity Linked Wealth- the value of equity held + the estimated market value of options held.","It can also be seen in FIG.","11 that the cells in the table are highlighted in a plurality of different colours, this feature gives an indication of the diversity of the data within the table and will be discussed in more detail hereafter.","The Remuneration Summary Bubble Chart, as shown in FIG.","12, is a chart that plots the three variables from the Remuneration Summary Table, i.e.","the Total Direct Remuneration, the Total Equity Linked Remuneration and the Total Equity Linked Wealth onto the same graph providing a simple a quick analysis method for the data.","The x-axis on the graph plots the Direct Remuneration value, and the y-axis plots the Total Equity Linked Remuneration values for each individual Director on the Board.","Each data point is then expanded in a circular form, wherein the diameter of each circle is proportional to the value of the Total Equity Linked Wealth of each Director.","The circles are colour coded to indicate the difference between different types of data, e.g.","executive directors, non-executive directors, averages, etc.",".",".",".","It can also be seen in FIG.","12 that various averages and ‘peer group’ averages are plotted, these will discussed hereafter.","The Direct and Equity Remuneration Bar Chart is a standard bar chart, with each Director placed along the x-axis, and the values of their respective Total Direct Remuneration and Total Equity Linked Remuneration for a particular financial period shown as bars, the size of which are relative to a scaled y-axis (for example in £000s of pounds).","Lines are also placed on the plot to indicate the values of the Director and peer group averages mentioned above.","The Total Equity Wealth Chart is another bar chart showing each Director along the x-axis with a bar extended upwards relative to the y-axis indicating the relative value of the Total Equity Wealth of each Director.","Again, some average values are also indicated.","The Direct Remuneration page provides a breakdown of the Total Direct Remuneration figures given in the Board Remuneration Summary, for a particular financial period.","The display is in the form of a table, with format similar to that of the Remuneration Summary in FIG.","11.The parameters displayed are: Executive Name- names of all of the Board Directors Role- The position held by the Director in the Company.","Salary- base annual pay Bonus- payments made in addition to salary Pension- employer's contribution to the employee's pension scheme Other- other ad hoc payments, such as relocation and fringe benefits Total Direct- sum of salary, bonus, pension and other income.","The Equity Linked Remuneration page provides a breakdown of the Total Equity Remuneration figures given in the Board Remuneration Summary, for a particular financial period.","The display is in the form of a table, with format similar to that of the Remuneration Summary in FIG.","11.The parameters displayed are: Executive Name- names of all of the Board Directors Role- position each Director holds on the Board.","Value of Equity Awarded- value of company shares awarded in the period.","LTIP Value- value of Long Term Incentive Plan equity contributions.","Intrinsic Value of Options Awarded- the amount by which the current stock price exceeds the exercise price of the options.","Note: if the stock price is below the exercise price, there is no intrinsic value.","Estimated Value of Options Awarded- Current estimate of the market value of Options gained, based on the latest closing stock price using the Black Scholes option pricing model, or a modification thereof.","Total Equity Linked Remuneration the sum of all of the for the Period- other equity linked variables in the table, i.e.","the sum of the variables in this list.","The Remuneration Ratios page is a series of ratios of some of the remuneration parameters mentioned above.","The display is in the form of a table, as shown in FIG.","13.The ratios displayed are: Executive Name- names of all of the Board Directors Role- The position held by the Director in the Company.","‘Bonus/(Bonus + Salary)’ Ratio ‘Equity Linked/Total Remuneration’ Ratio % Change the percentage change of an individual's Remuneration- remuneration from the one fiscal period to another, e.g.","from the ‘99 financial year to the ‘00 financial year.","Wealth Delta- the percentage increase in a Directors Wealth due to a 1% increase in a particular company's share price, i.e.","a measure of a Director's personal financial gain relative to the company share price success.","Board Characteristics The Board Characteristics section can be broken down into five main sections: Roles Network/Biographical Experience Change Diversity as can be seen in FIG.","5.These sections will now be discussed in turn.","The Board Characteristics ‘Roles’ page provides a breakdown of and parameters relating to the Role that each individual Director has in the company.","The page is displayed in a tabular form, similar to that shown in FIG.","25 (FIG.","25 is the Director Characteristics Roles page, but the format is very similar).","The parameters displayed on this Roles page are: Executive Name- names of all of the Board Directors Role- The position held by the Director in the Company.","Time to Retirement- The length of time to retirement, based on the company's policy retirement age (in years).","Shared External Directorships- The number of Directors on the subject Board who serve on other boards together.","Time in Role- number of years the Director has been in his/her current role on Board.","Time in Board- number of years the Director has been on the Board, in any role.","Time in Company- number of years under employment with company.","Time in Sector- number of years the Director has worked in the market sector.","The Board Characteristics ‘Network/Biographical’ page provides a breakdown of and parameters relating to the a Director's current Board level commitments, their history of working Board level commitments, and some basic personal details.","The page is displayed in a tabular form, as shown in FIG.","14.The parameters displayed on this Network/Biographical page are: Executive Name- names of all of the Board Directors.","Role- The position held by the Director in the Company.","Boards to Date Listed- the total number of Boards of public listed companies on which the Director has served to date including current Boards positions.","Boards to Date Unlisted- the total number of Boards of limited companies on which the Director has served to date including current Boards positions.","Boards to Date Other- the total number of Other Boards on which the Director has served to date including current Boards positions.","Current Boards Listed- the total number of Boards of public listed companies on which the Director currently serves.","Current Boards Unlisted- the total number of Boards of limited companies on which the Director currently serves.","Current Boards Other- the total number of Other Boards on which the Director currently serves.","Average Time in Other the average length of time spent by Companies- individual Directors in companies other than the subject company (in years).","Age- the age of the Director Education- the number of qualifications (degree level, inc. professional qualifications) held by the Director.","Gender- Shows either the gender of the Director or the proportion of the Directors that are male.","Nationality- individual nationalities for each Director, and a nationality mix ratio for the average values (where a value of 1 indicates the most diverse Board possible and a value of 0 indicates a Board comprising all directors of one single nationality).","The Board Characteristics ‘Experience’ section consists of two pages, as shown on FIG.","5, a Table and a Radar Chart, providing the same information in two different forms.","The pages provide a breakdown of and parameters relating to the Experience of the members of the Board, by mixing together some previously discussed parameters from other pages.","The pages are displayed firstly in tabular form, similar to that shown in FIG.","26, and in graphical form similar to that shown in FIG.","27 (FIGS.","26 and 27 are the Director Characteristics Experience pages, but the format is very similar).","The parameters displayed on the tabular page are: Executive Level- these figures are given for the executives collectively, unlike each individual Director's statistics as before.","This column defines between Executive Directors, Non-Executive Directors, and the Board as a whole.","Current Boards Listed- the total number of Boards of public listed companies on which the Director currently serves including current Boards positions.","Boards to Date Listed- the total number of Boards of public listed companies on which the Director has served to date including current Boards positions.","Time in Sector- number of years Director has worked in the market sector.","Time on Board- number of years a Director has served on the Board, in any role.","Age- the age of the Director Education- the number of qualifications (degree level, inc. professional qualifications) held by the Director.","The Board Characteristics Experience Radar Chart is a multi axis chart that will display the parameters displayed in the Experience Table page on one visual chart.","The radar charts will be discussed in more detail hereafter.","The Board Characteristics ‘Change’ section also consists of two pages, as shown on FIG.","5, a Table and a Radar Chart, providing the same information in two different forms.","The pages provide a breakdown of and parameters relating to the stability of the Board, by mixing together some previously discussed parameters from other pages and introducing some new parameters from them.","The pages are displayed firstly in tabular form, as shown in FIG.","15, and in graphical form as shown in FIG.","27 (FIG.","27 is the Director Characteristics Experience page, but the format is very similar).","The parameters displayed on the tabular page are: Executive Level- these figures are given for the executives collectively, unlike displaying each individual Director's statistics as in pages discussed earlier.","This column defines between Executive Directors, Non-Executive Directors, and the Board as a whole.","Attrition Rate- Proportion of Directors who have left in the previous year.","Succession Factor- the standard deviation of the time to retirement.","This measures the clustering of Directors and the proximity of Directors to retirement.","Standard Deviation of Time on standard deviation from the Board- average value of the time on the Board of all members of each executive group.","Average Time in Other standard deviation from the average Companies- value of the time in other companies for all members of each executive group.","Direct Remuneration- the average Direct Remuneration for each executive group.","Equity Linked Remuneration- the average Equity Linked Remuneration for each executive group.","The Change Radar Chart, as for the Radar Chart in the previous section, is a multi axis chart that will display the parameters displayed in the Change Table page on one visual chart.","These charts will be discussed in more detail hereafter.","The Board Characteristics ‘Diversity’ section also consists of two pages, as shown on FIG.","5, a Table and a Radar Chart, providing the same information in two different forms.","The pages provide a breakdown of and parameters relating to the diversity of the Board, by displaying the diversity of some previously discussed parameters from other pages.","The pages are displayed firstly in tabular form, as shown in FIG.","16, and secondly in graphical form as shown in FIG.","17.The parameters displayed on the tabular page are: Executive Level- these figures are given for the executives collectively, as with the change statistics above.","This column defines between Executive Directors, Non-Executive Directors, and the Board as a whole.","Standard standard deviation from the Deviation of average time served on the Board Time on Board- by all of the Directors in each executive group.","Standard standard deviation from the Deviation average time served in the of Time in company by all of the Company- Directors in each executive group.","Standard standard deviation from the Deviation average time served in the market of Time in sector by all of the Directors in Sector- each executive group.","Standard standard deviation from the Deviation of average number of Boards Listed Boards the Directors of each to Date- executive group have served on.","Standard standard deviation from the average age of Deviation the Directors of each executive group.","of Age- Standard standard deviation from the average Deviation number of qualifications gained by of Education- the Directors of each executive group.","Gender- the proportion of males in each executive group.","Nationality- a nationality mix ratio where a value of 1 indicates the most nationally diverse Board possible and a value of 0 indicates a Board comprising all directors of one single nationality.","The Diversity Radar Chart, as for the charts in the previous two sections, is a multi axis chart that will display the parameters displayed in the Change Table page on one visual chart, as shown in FIG.","17.These charts will be discussed in more detail hereafter.","Board Profile The Board Profile provides some general contact information for the subject company, e.g.","address, phone number, web site etc.","It also provides a short list of some basic financial data for the company, such as the Revenue Figures, Profit & Loss Figures and some key ratios, such as the P/E Ratio.","Thirdly it provides a list of advisers to the company, such as it's auditors, solicitors and financial advisors.","Director Summary The Director Summary, as shown in FIG.","22, provides a general overview of the positions held by a Director and displays the following basic information relating to those positions: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.","Time on Board- The length of time the Director has been on the Board (in years).","Time in Company- The length of time spent by the Director in the company (in years).","Time to Retirement- The length of time to retirement, based on the company's policy retirement age (in years).","Stakeholder Representative- specific stakeholder group(s) represented by the Director for example shareholders representatives, union, employees..","The second theme to the Director summary is a list of major shareholders in the company, giving their name and an indication as to the size of their holding.","Director Associations The Director Associations section has two levels beneath it, as shown in FIG.","6, named ‘Single and Multi Associations’, as shown in FIG.","23, and ‘Multi Associations’.","These two pages provide lists of associations that the Director has with other Directors through the Boards he/she serves on.","A ‘Single Association’ occurs when a Director on the Director of one particular company is also serving on the Director of another company, and is the only person from the subject company to have any association with that latter company.","A ‘Multi Association’ occurs when the Director sits on more than one Board with another Director.","Director Remuneration The Director Remuneration section can be broken down into six main sections: Remuneration Summary Direct Remuneration- annual cash based pay.","Equity Linked Remuneration- annual equity linked pay.","Equity Linked Wealth- total equity linked benefits received.","Remuneration Ratios Options, Pensions, and LTIP This hierarchy can be seen in FIG.","7.These sections will now be discussed in turn.","The Remuneration Summary, as shown in FIG.","7, consists of four separate pages, each displaying the summary information in a different manner.","The Remuneration Summary Table, which is similar to that shown in FIG.","11 (FIG.","11 is the Board Remuneration Summary, but the format is very similar), displays the following parameters: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.","Total Direct Remuneration- amount of cash based remuneration, for a particular financial period (e.g.","a financial year).","Total Equity Linked Remuneration- amount of remuneration in the form of equity, shares, and options for a particular financial period.","Total Remuneration for the Period- the sum of the Total Direct Remuneration and the Total Equity Linked Remuneration, each as described above.","Total Equity Linked Wealth- the value of equity held + the estimated market value of options held.","As with the Board Remuneration Summary Table in FIG.","11, the Director data tables are highlighted in accordance with the diversity of the data, this will be discussed hereafter.","The Remuneration Summary Bubble Chart, similar to that shown in FIG.","12 (FIG.","12 is the Board Remuneration Summary Bubble Chart, but the format is very similar), is a chart that plots the three variables from the Remuneration Summary Table, i.e.","the Total Direct Remuneration, the Total Equity Linked Remuneration and the Total Equity Linked Wealth onto the same graph providing a simple a quick analysis method for the data.","The x-axis on the graph plots the Direct Remuneration value, and the y-axis plots the Total Equity Linked Remuneration values for each position that the Director holds.","Each data point is then expanded in a circular form, wherein the diameter of each circle is proportional to the value of the Total Equity Linked Wealth of the Director in each position held.","The circles are colour coded to indicate the difference between different types of data, e.g.","executive Director positions, non-executive Director positions, averages, and ‘peer group’ averages.","The Direct and Equity Remuneration Bar Chart is a standard bar chart, with each Director position placed along the x-axis, and the values of their respective Total Direct Remuneration and Total Equity Linked Remuneration for a particular financial period shown as bars, the size of which are relative to a scaled y-axis (for e.g.","in £000s of pounds).","Lines are also placed on the plot to indicate the values of the Director and peer group averages mentioned above.","The Total Equity Wealth Chart is another bar chart showing each Director along the x-axis with a bar extended upwards relative to the y-axis indicating the relative value of the Total Equity Wealth of each Director.","Again, some average values are also indicated.","The Direct Remuneration page provides a breakdown of the Total Direct Remuneration figures given in the Director Remuneration Summary, for a particular financial period.","The display is in the form of a table, with format similar to that of the Board Remuneration Summary in FIG.","11.The parameters displayed are: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.","Salary- base annual pay Bonus- payments made in addition to salary Pension- employer's contribution to the employee's pension scheme Other- other ad hoc payments, such as relocation and fringe benefits Total Direct- sum of salary, bonus, pension and other income.","The Equity Linked Remuneration page provides a breakdown of the Total Equity Remuneration figures given in the Director Remuneration Summary, for a particular financial period.","The display is in the form of a table, with format similar to that of the Board Remuneration Summary in FIG.","11.The parameters displayed are: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.","Value of Equity Awarded- value of company shares awarded in the period.","LTIP Value- value of Long Term Incentive Plan equity contributions.","Intrinsic Value of Options Awarded- Current Realisable Profit on Options awarded.","Estimated Value of Options Awarded- Current estimate of the market value of Options gained, based on the latest closing stock price using the Black Scholes option pricing model.","Total Equity Linked Remuneration the sum of all of the for the Period- other equity linked variables in the table, i.e.","the sum of the variables in this list.","The Remuneration Ratios page is a series of ratios of some of the remuneration parameters mentioned above.","The display is in the form of a table, as shown in FIG.","24.The ratios displayed are: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.","‘Bonus/(Bonus + Salary)’ Ratio ‘Equity Linked/Total Remuneration’ Ratio % Change Remuneration- the percentage change of an individual's remuneration from the one fiscal period to another, e.g.","from the ‘99 financial year to the ‘00 financial year.","Wealth Delta- the percentage increase in a Directors Wealth due to a 1% increase in a particular company's share price, i.e.","a measure of a Director's personal financial gain relative to the company share price success.","The Remuneration Options, Pensions and LTIP page provides a revue of these parameters.","Director Characteristics The Director Characteristics section can be broken down into three main sections: Roles Network/Biographical Experience as can be seen in FIG.","8.These sections will now be discussed in turn.","The Director Characteristics ‘Roles’ page provides a breakdown of and parameters relating to each Role held by the Director.","The page is displayed in a tabular form, as shown in FIG.","25.The parameters displayed on this Roles page are: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.","Time to Retirement- The length of time to retirement, based on the company's policy retirement age (in years).","Shared External Directorships- The number of Directors who serve on other Boards with the Director.","Time in Role- number of years the Director has been on his/her current role on Board.","Time on Board- number of years the Director has been on the Board, in any role.","Time in Company- number of years under employment with company.","Time in Sector- number of years the Director has worked in the market sector.","The Director Characteristics ‘Network/Biographical’ page provides a breakdown of and parameters relating to the a Director's current Board level commitments, their history of working Board level commitments, and some basic personal details.","The page is displayed in a tabular form, similar to the Board Characteristics Network/Biographical page as shown in FIG.","14.The parameters displayed on this Director Network/Biographical page are: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.","Boards to Date Listed- the total number of Boards of public listed companies on which the Director has served to date including current Board positions.","Boards to Date Unlisted- the total number of Boards of limited companies on which the Director has served to date including current Board positions.","Directors to Date Other- the total number of Other Boards on which the Director has served to date including current Board positions.","Current Boards Listed- the total number of Boards of public listed companies on which the Director currently serves.","Current Boards Unlisted- the total number of Boards of limited companies on which the Director currently serves.","Current Boards Other- the total number of Other Boards on which the Director currently serves.","Average Time in Other the average length of time spent by Companies- individual Directors in companies other than the subject company (in years).","Age- the age of the Director Education- the number of qualifications (degree level, inc. professional qualifications) held by the Director.","Gender- Shows the gender of the Director.","Nationality- shows the nationality of the Director.","The Director Characteristics ‘Experience’ section consists of two pages, as shown on FIG.","8, a Table and a Radar Chart, providing the same information in two different forms.","The pages provide a breakdown of and parameters relating to the Experience of the Director, by mixing together some previously discussed parameters from other pages.","The pages are displayed firstly in tabular form, similar to that shown in FIG.","26, and in graphical form as shown in FIG.","27.The parameters displayed on the tabular page are: Executive Name- names of the companies with whom the Director has a Board level position Current Boards Listed- the total number of Boards of public listed companies on which the Director currently serves including current Board positions.","Boards to Date Listed- the total number of Boards of public listed companies on which the Director has served to date including current Board positions.","Time in Sector- number of years Director has worked in the market sector.","Time on Board- number of years a Director has served on the Board, in any role.","Age- the age of the Director Education- the number of qualifications (degree level, inc. professional qualifications) held by the Director.","The Director Characteristics Experience Radar Chart is a multi axis chart that will display the parameters displayed in the Experience Table page on one visual chart.","The radar charts will be discussed in more detail hereafter.","Director Profile The Director Profile provides some general personal information for the Director, e.g.","date of birth, nationality etc.","It also provides a short CV showing the positions that he/she currently holds and positions that the Director has held in the past, with a brief description of each of those positions.","User Interface Having looked at the structure of the information pages displayed on the user's web browser, the method by which the user navigates around these pages and how they select which data is to be displayed will now be described.","FIG.","9 shows an example of the general layout of the user interface.","The display comprises two parts: a fixed thin navigation pane on the left hand side of the display which displays the list of available information page windows that a user can select, these were discussed above, as well as other features such as a search facility and some historical selection lists, that will be discussed hereafter; the second part of the display comprises a changeable information display pane which can display a series of web style pages that each provide a different set of the actual Board/Director related information pages that are received from the web server.","A user can select which page of data he wishes to have displayed in the right hand pane by selecting from the hierarchical list in the left hand navigation pane.","The navigation pane provides a lower level break down of available pages as the user navigates around the system.","For instance in FIG.","9 the top level structure of the system, as shown in FIG.","2, is visible, whereas in FIG.","11, where the user has selected the ‘Board Remuneration Summary’ screen, it can be seen that the navigation list has been extended to show the levels below that of the screen which is being displayed, hence allowing the user to access different methods of display for the ‘Board Remuneration Summary’ information; ‘Table’, ‘Bubble Chart’, ‘Direct and Equity Chart’ and ‘Equity Wealth Chart’ in this case.","Once the user has selected either the Board section or the Director section, by clicking on one of the two labelled tabs in the top left hand corner of the screen, they have to next choose a particular Board/Director to be used as the subject of the data.","This selection will instruct the system to download all relevant information for that Board/Director to then allow the user to navigate around the different screens in the navigation panel viewing different data sets related to that Board/Director.","This Board/Director selection is conducted by using one of two search systems which are situated in the navigation pane.","FIG.","9 shows these search facilities situated below the screen navigation list in the navigation pane on the left hand side of the display.","One search option, the first box below the screen navigation list, comprises a manual entry search system which allows the user to enter some data which the system then uses to search the database.","Another search option is called the ‘Advanced Search’ and is situated in the box below the manual search data entry boxes, this involves the user defining a search by selecting from a number of parameters in a pre-defined list that the system will use to narrow the search of the database.","The manual search facility offers two methods of data entry for the search; a user can either input a company/person's name into the ‘Company Name’ text box displayed on the navigation pane; or the user can input another form of company related designation data, such as the SEDOL CODE—a financial markets company ID (the data type can be selected from a drop-down box), by inputting data into the ‘Company ID’ text box on the navigation pane.","The system will then conduct a search of the database and produce a search results list such as those shown in FIGS.","21 and 31.The user can then select any Board/Director he wishes to use as the subject of the analysis in the information display pages by clicking on a Board/Director's name from the search results list.","The system will then retrieve the parameters for that chosen Board/Director and initially display the ‘Summary’ page for that subject Board/Director, as shown in FIG.","3.The Advanced Search facility is initiated when the user clicks on the ‘Advanced Search’ button in the navigation pane.","A screen such as that in FIG.","20 will be displayed for an advanced Board search, and such as that in FIG.","30 for and advanced Director search.","The user will then select at least one of the pre-defined parameters, such as the country or sector of the Board/Director they wish to search for and/or by inputting at least one piece of data into any one of the data entry fields, such as defining a Market Capitalisation of the company of the Board they wish to search for.","The system will then conduct a search of the database based on the data entered by the user and produce a search results list, again such as those shown in FIGS.","21 and 31.The user can then select any Board/Director he wishes to use as the subject of the analysis in the information display pages by clicking on a Board/Director's name from the search results list, the system will then retrieve the parameters for that Board/Director and initially display the ‘Summary’ page for that subject Board/Director, as shown in FIG.","9.A third method of choosing a subject Board/Director to be the subject of the information pages is to select a name from the ‘Viewed Boards’ list at the bottom of the navigation pane.","The system will store a list of the last n subjects to have been chosen by the user and display them at the bottom of the navigation pane.","This allows the user to easily return to a recent set of displayed data without having to pass through either of the two search methods again.","Once a subject Board/Director has been chosen and its data downloaded and displayed, the name of that Board/Director will be displayed in the top left-hand corner of the display on all of the information pages.","When a new Board/Director is chosen, the name displayed will be updated accordingly.","Many of the information pages display the information in tabular form, and there is a marked difference between the style of these tables for the Board section and the Director section in terms of how the data is displayed, these two different styles will now be explained, firstly with reference to Boards.","The rows of the Board related tables of data are split up into three main categories, the top category comprising the Executive Directors, the second category comprising the Non-Executive Directors and the third category comprising All Positions.","FIG.","11 shows how this structure is displayed.","The top category of FIG.","11 lists the names of the Executive Directors of ‘Barclays’, the second category lists the names of all of the Non-Executive Directors of Barclays, and the bottom category contains the average values for all of the Directors together.","It should be noted that this format allows the user to analyse the data for every individual on the selected subject Board, as well as the average value for the ‘executives’ category, an average value for the ‘non-executives’ category and the average value for all members of the Board collectively.","Thus, this set-up provides a large amount of segregated and averaged data to allow the user to analyse a detailed view of all aspects of a Board at the same tine.","The Director section display is shown in FIG.","25.It can be seen that the selected Director is Tessen von Heydebreck and that he serves on two Boards, as an Executive Director with Deutsche Bank, and a Non-Executive Director with BASF AG.","In a similar manner to the Board tables, these different positions are split in to separate categories.","Any Executive positions will be displayed in the top category, and any Non-Executive positions in the second category, with an average for all positions held shown in the bottom category.","Hence, all aspects of an individual Director's commitments are displayed at one time, as well as an average for those positions, providing a comprehensive analysis platform without having to look up each position individually.","Peer Groups One of the features of the embodiment of the present invention are the so called ‘Peer Groups’ that allow the user to compare a selected subject Board/Director with one or more corresponding Boards/Directors.","These Peer Groups act as a benchmark with which to compare and contrast different Boards/Directors, they can comprise one single Board/Direct to allow a direct comparison between two particular companies/individual Directors, or they can contain a plurality of different Boards/Directors, grouped for example by sector.","As mentioned above, the tables of data for both the Board and Director sections are separated into three categories, and as well as showing data for the individual Directors of a Board or the individual Positions held by a Director, each of these categories also displays an average value for all data within that category.","Below these average values, as can be seen in FIGS.","11 and 25, is displayed an average value for all of the members of a Peer Group, this can be used as a comparative measure for the individual Board members/Director positions, as well as for the average values for each category.","A key function of these Peer Groups is to enable the user to define the constituents of any Peer Group and can select any Peer Group from their records to be used in the comparison.","At the bottom of the navigation pane, above the Viewed Boards/Directors section is the Peer Group Button which when selected opens up the Peer Group management screens.","These screens work in exactly the same way for the Board sections as for the Director section and will therefore be explained together, with reference to a number of corresponding diagrams.","FIGS.","18 and 28 show the first page of the Peer Group management pages.","This page displays all of the Peer Groups that have been set up by the user to date.","We can see in FIG.","18 that the user has set up three Peer Groups: PG1, PG2 and PG3, and that those Peer Groups contain seven, three and two members respectively.","From the Peer Group amendment screen the user is able to view the members of any Peer Group, amend the members of any Peer Group, or set up a new Peer Group.","To view the members of a Peer Group, shown in FIGS.","18 and 28, the user simply clicks on the name of one of the Peer Groups and a screen containing a list of Peer Group members is displayed, such as those shown in FIG.","19 for the Board Peer Group PG1 and in FIG.","29 for the Director Peer Group PG2.Hence it can be seen that the Board Peer Group PG1 consists of seven Boards, including those of Barclays and Marks & Spencer, and the average parameter values displayed on a table of data on a Board information screen, for this Peer Group, would be equal to the average value for all of the groups in the Peer Group.","The user can easily amend the members of a Peer Group whilst in the Peer Group members screen, as in FIG.","19 or 29, by adding a new member or deleting an existing Peer Group member, as well as being able to move or copy a member to another Peer Group.","To delete a member the user would check the check box to the left of any current Peer Group member and then click on the delete button in the top left hand corner of the screen.","To copy or move a member the same process would be used except that having selected a member to copy or move the user would choose another Peer Group from a drop-down menu in the top right corner, once a member and other Peer Group have been chosen the user clicks either the copy or move button and the action is completed.","Adding a new user to a Peer Group is slightly more complicated and requires the use of the two searching tools, manual and advanced, that were discussed earlier.","A user can search as described earlier by company name, company ID or by using the advanced search and a search results page is displayed, as in FIGS.","21 and 31.If the Board/Director is present in the search result list, the user will check the check box next to the displayed name and then select a Peer Group from a drop-down menu in the top right hand corner of the screen into which they wish to place Board/Director.","Once a subject and Peer Group have been chosen, the user clicks on the ‘Add To Peer Group’ button above the search results list and the action is duly performed.","Setting up a new Peer Group mainly involves a combination of the features that have already been discussed, once the initial naming procedure has been carried out.","In the main Peer Group page, FIGS.","18 and 28, the user can set up a shell for a new Peer Group by typing a name into the text box next to the ‘New’ button above the list of current Peer Groups.","Once a name has been entered and the ‘New’ button has been selected, the shell is created and the name of the new Peer Group are displayed along with the other current Peer Groups.","The user can then enter that new Peer Group's member screen and add new members, or indeed copy/move members from other group's member screen to the new Peer Group.","Once one or more Peer Groups have been set up, the user is able to use the Peer Group comparison facility in one of the main Board/Director information screens.","Most of the tabular and graphical screens contain a combination of a series of drop-down menus in the top right hand corner of the screen.","These enable the user to perform one of three actions: choose a Peer Group to act as a comparison; choose and individual member of any one Peer Group to act as a comparison, or add the current subject Board/Member to a Peer Group.","When the user selects a Peer Group from the ‘Compare to Peer Group’ drop-down list, the system will display the average value for all members of that Peer Group for all of the parameters in any information table or graph.","For instance, in FIG.","14, the Compare to Peer Group display box in the top right hand corner of the screen is indicating that Peer Group PG1 has been selected for the comparison to Nokia OYJ, the subject Board, named in the top left hand corner of the screen.","The Peer Group averages in the table refer to the average value of all of the members of Peer Group PG1.If the user wished to compare Nokia against a different Peer Group they are able to select a different Peer Group name from the Compare to Peer Group selection drop down menu.","The user is able to compare the subject Board, Nokia OYJ in the example in FIG.","14, against a particular member of the selected Peer Group, PG1 in this example, by selecting one of the members of PG1 from the ‘Compare to Peer Member’ drop-down list, below the ‘Compare to Peer Group’ list in the display.","The user could, for instance, choose ‘Marks & Spencer's’ from the PG1 Peer Group list, this would then display the average values for the Marks & Spencer's Board on the table of data and hence allow the user to compare the Boards of Nokia OYJ with Marks & Spencer's directly.","The third drop-down menu in the top right hand corner of the screen allows the user to add the current subject Board, Nokia OYJ in the example of FIG.","14, to any current Peer Group.","The user simply clicks on the drop-down menu and a list of existing Peer Groups is displayed, the user can then select one of these per groups and the subject Board is added to that Peer Group automatically.","The Peer Group system works in exactly the same manner for the Director information screen as explained above for the Board information screens.","The only difference being that the Peer Groups are made up of individual Directors and the ‘Compare to Peer Member’ list will offer a list of said individuals to allow the user to compare the subject Director to any one specific other Director.","It should be noted that the Peer Group comparisons are also use in the graphical information pages.","For instance their values are plotted on the bubble charts for comparison purposes, as mentioned earlier.","Also, they are used as a comparative means on the radar charts, FIG.","17 shows an example of this where the thin line is that of the selected Board, Royal Bank of Scotland, and the shaded area represents the values for the selected Peer Group PG1.Again these pages have the functionality to change the Peer Group, view an individual member of a Peer Group or add the subject Board to a Peer Group.","Colour Coding Another feature of the embodiment of the current invention, as mentioned earlier, is the highlighting of the cells of the tables of information to visually indicate the diversity of the data.","Again the Peer Groups play an important part in this function as the reference point from which the diversity is calculated.","FIG.","14 shows a table of data for the Network/Biographical parameters.","Displayed above the table is a ‘Variance key’ that indicates the association between the colour of the highlighting of each cell and the measure of variance of the data in the cell, as compared to its corresponding Peer Group average value.","The cells are highlight in relation to the number of standard deviations that the data contained therein is away from the average value of the selected Peer Group.","If the data is within ±1 standard deviations of the Peer Group average, the cell will not be highlighted.","If the cell is between −1 and −2 or +1 and +2 standard deviations of the average of the Peer Group it is highlighted in a light colour, and so on where the highlighting becomes darker with the more standard deviations the data is away from the Peer Group average.","This feature provides easily recognisable information to a user.","Patterns and trends can be identified in the data much more easily using this feature.","It removes the need for a user to laboriously read every data parameter in the tables in order to analyse the data.","Radar Charts Another feature of the embodiment of the current invention is the radar charts, and especially the method used to determine the scale of the axes.","FIG.","17 shows a Diversity Radar Chart for the Royal Bank of Scotland and Peer Group PG1.This radar chart is the corresponding graphical representation of the data in the Diversity Table in FIG.","16.The first aspect to note is the number of columns of data within the Diversity Table and the corresponding number of axes on the Diversity Chart.","The chart will display as many parameters from the table as it has data for.","In many cases the Diversity table is fully populated with data and the Diversity Chart will have a potential 8 axes.","However, if there is insufficient data available, i.e.","there is no data for the subject Board or there is no data for the selected Peer Group, the radar chart is adjusted and the axis related to the parameter with insufficient data is removed and hence not displayed.","It can be seen from FIGS.","16 and 17 that there is only sufficient data in four of the parameter columns of the Diversity Table, the Gender column does contain some data but is not fully populated, hence the corresponding Radar Chart only has four axes.","The scaling of the axes is based on a similar method to that of the highlighting mentioned above, in that they are bound by a range of standard deviations.","The axes are bound by a range of ±3 standard deviations from the average of a Peer Group.","Hence the minimum value of each axis on the Radar Chart equals −3 standard deviations from the average of a Peer Group and the maximum value of each axis equals +3 standard deviations from the average of a Peer Group.","In this particular embodiment of the invention the Peer Group used for this scaling is not the user selected Peer Groups, it comprises all of the Boards/Directors that are based in the same country as the subject Board/Director.","For example in FIG.","17, the Royal Bank of Scotland is based in the UK, hence the scale of the axes in the corresponding Diversity Chart are ±3 deviations from the average value for all other Boards on the Database that are also based in the UK.","This feature provides a useful method of displaying the data to allow easy analysis by a user.","The multi axis plot facilitates comparative analysis of a multitude of parameters related to Boards and/or Board members.","Date Change Another feature of the embodiment of the invention is the date change facility.","On each information display page there is a date in the top left corner, below the subject Board/Director name, as can be seen in FIGS.","16 and 17 for example.","The date displayed in the box denotes the point in time to which the information displayed relates.","The user is able to change this date, by selecting another date from the drop down menu, hence allowing them a very simple tool to view historical data across a wide date range.","Mouse Over Another feature of the embodiment of the invention is the ‘mouse-over’ facility.","The system displays small information notes next to a mouse pointer as the user moves around the screen to enable the user to quickly understand the functionality of a particular feature on the display.","Additional to this is a mouse-over feature to aid the colour coding of data tables facility discussed above.","As a user passes their mouse over a shaded cell in a table of data a small information note is displayed showing the specific value of deviation that the particular parameter has from the Peer Group average value, this feature adds more detail to the colour coding system discussed above, when it is required by the user.","Pre-Defined Peer Groups In another embodiment of the current invention a plurality of pre-defined Peer Groups are be loaded onto the database.","The user is able to use, select, and even amend, these pre-defined Peer Groups in the same way as the current user defined Peer Groups, discussed above.","These pre-defined Peer Groups are organised into logical groups, such as by sector or geographic region and allows the user to compare a subject Board/Director against one of these default Peer Groups without having to set-up a Peer Group themselves.","Fantasy Board In another embodiment of the current invention the Board and Director sections are mixed together and function as one unit displaying data for both individual Directors as well as Boards in the same information tables and on the same information charts.","This allows for analysis of Boards against Directors, e.g.","a user can set up a ‘fantasy Board’ by setting up a Peer Group of Directors and then compare that fantasy Board against a real Board.","Or, in turn, an individual Director can be compared against the average value for a Peer Group of one or more Boards.","Enhanced Board/Member Associations Functionality In another embodiment of the current invention the Board/Director Associations information pages will not only display links between the members of the subject Board/subject Director and other positions that they may hold, but also associations from a subject Board/Director to a target Board Director, via a plurality of other Boards/Directors Associations.","For Example, this will allow the user to input their subject Board/Director and a ‘target’ Board/Director with which they would like to make contact.","The system will search the database and display a chain of associations which link the two user selected Boards/Directors together.","This display may provide a search result such as: Board 1 is linked to Board 3 via Mr. A; Board 3 is linked to Board 5 via Mr. B; Board 5 is linked to Board 2 via Ms. C; Where Board 1 is the ‘subject’ Board and Board 2 is the ‘target’ Board.","This functionality allows the user to build networks of Boards and Directors for use in a plurality of business situations.","Company Advisors Associations In another embodiment of the current invention, the associations functionality is extended to encompass the advisors of companies, a list of which is already provided on the Board Profile information page for each Board.","In this embodiment a user is able to conduct a search of Associations based on a particular advisor, i.e.","make the advisor the subject of the information and display the associations for that advisor.","For example, if a particular company has been in trouble as a result of one of their advisors, a user can conduct a search of the Associations of that advisor to other Boards and the system would display a list of any other Boards that use the same advisor.","This is useful, for example in the field of fund management, where an advisor has recently shown weakness whilst acting for one of a Fund Manager's holdings, that fund manager could perform a due diligence check to ascertain if any of his other shareholdings are advised by the same advisor by conducting an Associations search on that advisor.","Board Summary ‘Timeline’ In another embodiment of the current invention a ‘Board Timeline’ chart, as that shown in FIG.","32, is displayed on a second information display page under the ‘Board Summary’ section.","This information page displays a bar chart style chart over a discrete time period, pre-set or user defined, showing bars to represent the periods during which each Director has been on the Board, and the position they held, during the said time period.","Director Associtaion ‘Multi’ In another embodiment of the current invention a graphical bar chart style plot is displayed to provide a more user friendly method of viewing the Director Multi Associations.","An example of this graphical information page is in FIG.","37.The names of each Director are placed along the y-axis of a bar chart, with time (in years) along the x-axis.","A bar is then plotted alongside each Director's name for a period in which they held a Director level role, colour coding is used to differentiate between the different companies on which they served.","This chart provides a much more user friendly method of reviewing the multi associations of a number of Directors.","Analyses Topic Section In another embodiment of the current invention the Board and Director sections comprise a sixth main topic heading, in addition to the five existing main sections as discussed above and shown in FIG.","2, named ‘Analyses’.","This extra section is added to the user interface navigation pane, as shown in the navigation pane on the left hand side of FIGS.","32 and 37, between the Characteristics and Profiles sections.","The two sections contain different sub-layers, and will now be discussed in turn, beginning with the Board section.","Board Analyses The Board Analyses section can be broken down into five main sections: Board Share Price History Board and Shareholder Returns Board Time Analysis Board Experience over Time Options and Equity Scenarios Each of these sections will now be discussed in turn.","Board Analyses ‘Board Share Price History’, as shown in FIG.","33, displays the share price of the subject company over a discrete time period, pre-set or user-defined, with major Board related corporate actions indicated graphically on top of the chart.","For example, if a CEO resigns, a mark will be placed on the graph to indicate the point in time when this occurs.","This data can be loaded onto the database automatically or manually, as discussed above, via the analytics data entry method on the Data Repository System (1, FIG.","1) at the Central Data entry Management Location (100, FIG.","1).","This facility enables the user to analyse the fluctuations in the company share price relative to such Board related corporate events and allows them to gain a fuller understanding of share price volatility at such points in time.","Hence this embodiment offers the user a tool to assess the importance of Board stability for a particular company and hence gain an insight into future share price fluctuations on the announcement of a Board related corporate action.","Board Analyses ‘Board and Share Holder Returns’, as shown in FIG.","34, displays a mix of different data types on one graphical display, including Director Direct Remuneration, Director Equity remuneration, Company Income and the Company Share Price.","The main element of the graph is a bar chart with two bars for each year along the x-axis, one of these bars displays information relating to the remuneration (further broken down into sections of: salary, other direct, equity, options and LTIP) of the Directors of the Board during that year, the other showing the Net Income of the Company in the same year.","On top of the bar chart is displayed the share price of the subject company over the discrete time period, pre-set or user-defined, displayed.","This graph enables the user to analyse a number of different features at once, for instance the relationship between company net income and the share price, or that of the Directors income in relation to the share price, plus many more.","Board Analyses ‘Board Time Analysis’, as shown in FIG.","35, graphically displays a summary of the external commitments of the members of a Board.","The graph comprises a bar chart with three bars displayed in each year along the x-axis, each bar to represent board-level positions held on ‘listed’, ‘unlisted’ and ‘other’ boards respectively.","The y-axis provides a measure of the average number of those board-level positions held by the members of the subject board during each particular year.","This graph is a graphical representation of some of the information given in the Board Characteristics ‘Network/Biographical’ section discussed earlier.","Board Analyses ‘Board Experience over Time’, as shown in FIG.","36, displays a bar chart offering a summary of the experience of the members of the Board on a yearly time basis.","Each year plotted along the x-axis contains two bars: the first of which provides an average value, for all of the Board members, of the time spent in the current company, and a measure of how much of that time was spent serving on the Board; the second bar represents the average number, for all the members of the Board, of all the Boards on which they currently serve and all the Boards on which they have served to date.","This is another graphical representation related to the experience parameters displayed in the Board Characteristics ‘Experience’ Table information page.","Board Analyses ‘Options and Equity Scenarios’ allows the user to assess the change in the value of the Board held options and equity by running scenarios related to the stock price of a company.","The user can enter parameters for an imaginary rise or fall in the price of a stock and the system will calculate and display the effect that this change in stock price would have on the option and equity valuation of the Board of that company.","Director Analyses The Director Analyses section can be broken down into two main sections: Director Share Price History Options and Equity Scenarios These sections will now be discussed in turn.","Director Share Price History, as shown in FIG.","38, is a tool to allow the user to assess the impact of a Director on the company share price during each of his Director level positions.","A discreet time period, pre-set or user selected, is used and a separate share price chart for each of the companies on whose Board the Director has served during that time are arranged on the same screen.","On top of these share price graphs will be displayed time related information relating to when the Director served with each company and the corresponding role which he held on each Board.","This data, as with the Board time line analysis data is entered onto the system using the manual analytics software package on the Data Repository System (1, FIG.","1) at the Central Data entry Management Location (100, FIG.","1).","By reviewing the information displayed the user is able to quickly gain an impression of the historical impact of the arrival and departure of the Director in question at each of his past companies, as well as the performance during his/her time on the Board.","Hence, the system is able to provide an indicative historical performance analysis for the Director.","Director Analyses ‘Options and Equity Scenarios’ allows the user to assess the change in the value of the Director held options and equity by running scenarios related to the stock price of a company.","The user can enter parameters for an imaginary rise or fall in the price of a stock and the system will calculate and display the effect that this change in stock price would have on the option and equity valuation of a Director of that company.","Peer Group Alerts In another embodiment of the current invention The Peer Group Alerts section forms part of the Board Peer Groups Section discussed above.","These alerts target allow the user to receive targeted corporate action based information about a Board and/or Director Peer Group as follows: Board Peer Group Alerts allows the user to define a list of criteria for all the members of a Peer Group of Boards, on the action of which an alert will be flagged up to the user indicating the event that takes place.","FIG.","39 shows an example of the definitions screen, it can be seen that there is a wide range of Director level and Board level criteria which can be defined for the members of the Peer Group that has been selected from the drop-down at the top of the screen.","If any one of the pre-defined variables is met, for instance a Chairman of one of the Boards within the Peer Group leaves, or the average age of the Boards in the Peer Group rises above the selected level, then a message indicating the event will be supplied to the user, either by e-mail or on login depending on the user's preference, as shown in FIG.","40.This facility allows the user to target the corporate news that they require.","Instead of sifting through large quantities of literature to find the information that is useful to them, the user can define the areas in which they are interested, by using the screen shown in FIG.","39, for the Boards in which they are interested, by setting up a Peer Group containing those Boards, as discussed above, and hence receive news that is personally targeted at the user.","Director Peer Group Alerts allows the user to define a list of criteria for all the members of a Peer Group of Directors, on the action of which an alert is flagged up to the user indicating the event that takes place.","FIG.","41 shows an example of the definitions screen, it can be seen that there is a wide range of Director level criteria which can be defined for the members of the Peer Group that has been selected from the drop-down at the top of the screen.","If any one of the pre-defined variables is met, for instance a Director within the Peer Group leaves his position, or the average Direct Compensation within the Peer Group rises above the selected level, then a message indicating the event is supplied to the user, either by e-mail or on login depending on the user's preference, as shown in FIG.","42.This facility allows the user to target the corporate news that they require.","Instead of sifting through large quantities of literature to find the information that is useful to them, the user can define the areas in which they are interested, by using the screen shown in FIG.","41, for the Directors in which they are interested, by setting up a Peer Group containing those Directors, as discussed above, and hence receive news that is personally targeted at the user.","Although the present invention has been described hereinbefore with reference to specific embodiments, it will be apparent to a skilled person that modifications lie within the spirit and scope of the present invention.","For example, although the system has been described as being executed on a network, it could be implemented on a single computer or any suitable computer system."]],"string":"[\n [\n \"Company board data processing system and method\",\n \"A company board data processing system and method provides a user interface platform to supply information related to members of boards of companies to enable comparison of board members or boards at board member level.\",\n \"Parameters related to board members of a plurality of boards are stored.\",\n \"A used interface is generated which comprises a display of the parameters related to board members for a plurality of boards and/or individuals board members to allow comparison of board members and/or boards at boards member level.\",\n \"The system also allows the user to define a group of boards and/or board members, wherein processed parameters for said group can be used for comparison purposes.\"\n ],\n [\n \"1.A company board data processing system comprising: a board member store storing parameters related to board members of a plurality of boards organised into boards; and a user interface generator for generating a user interface displaying said parameters related to board members for a plurality of boards and/or individual board members to allow comparison of boards, at board member level, and/or board members.\",\n \"2-197.\",\n \"(canceled) 198.A company board data processing system as in claim 1, wherein said user interface generator is adapted to display said parameters for each member of at least one board and/or an average value of one or more of said parameters of said members of at least one board.\",\n \"199.A company board data processing system as in claim 1, further comprising a board parameter processor for processing parameters for a group of boards or board members to determine processed parameters comprising at least one of one or more parameter totals, one or more parameter averages and one or more parameter deviations, wherein said user interface generator is adapted to generate a user interface to display at least one of said processed parameters for at least one group of boards or board members to allow comparison with at least one board or board member.\",\n \"200.A company board data processing system as in claim 1, wherein said user interface generator is adapted to generate said user interface to display processed parameters comprising at least one of one or more totals, averages and deviations of said parameters related to a group of board members or boards to allow comparison with at least one board or board member.\",\n \"201.A company Board data processing system as in claim 1, including a ranking processor for ranking members of a group of boards and/or board members based on relative value of said parameters; wherein said user interface generator is adapted to generate a user interface to display said parameters of said boards and/or board members of said group to indicate ranking.\",\n \"202.A company Board data processing system as in claim 199, including a ranking processor for ranking groups of boards and/or board members based on relative value of said processed parameters; wherein said user interface generator is adapted to generate a user interface to display said processed parameters of said groups of boards and/or board members to indicate ranking.\",\n \"203.A company data processing system as in claim 199, wherein said group of board members comprises a user selected and/or pre-defined group of boards or board members assigned as a peer group.\",\n \"204.A company board data processing system as in claim 199, further comprising: user selector means for receiving a user selection of at least one board and/or board member for inclusion in a peer group; wherein said group of boards or board members comprises a user selected group of boards or board members assigned as a peer group; said parameter processor is adapted to process parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; and said user interface generator is adapted to generate a user interface displaying at least one of said processed peer group parameters.\",\n \"205.A company board data processing system as in claim 204, wherein said user selector means is arranged to receive a user selection of at least one board for comparison with at least one board peer group, and said user interface generator is adapted to generate a user interface to display the parameters related to members of said at least one selected board, to allow the or each board to be the subject of analysis.\",\n \"206.A company board data processing system as in claim 204, wherein said user interface generator is adapted to generate a user interface to display the parameters related to individual members of said at least one selected board.\",\n \"207.A company board data processing system as in claim 204, wherein said user selector means is arranged to receive a user selection of at least one board member for comparison with said board member peer group, and said user interface generator is adapted to generate a user interface to display the parameters related to said at least one selected board member, to allow said at least one selected board member to be the subject of analysis.\",\n \"208.A company board data processing system as in claim 204, further comprising a peer group data store storing data for one or more user selected peer groups; wherein said user interface generator is adapted to generate a user interface displaying a list of existing peer groups based on said stored peer group data; said user selector means is arranged to receive a user selection of one or more peer groups from said list; and said user interface generator is adapted to generate a user interface displaying at least one of said processed parameters for at least one of said selected peer groups.\",\n \"209.A company board data processing system as in claim 204, further comprising a comparison group amendment means allowing a user to amend the constituents of a peer group.\",\n \"210.A company board data processing system as in claim 209, wherein said peer group amendment means is adapted to allow a user to copy one or more constituents of a peer group to another peer group.\",\n \"211.A company board data processing system as in claim 204, further comprising a searching means arranged to allow a user to search for a particular board in the board member store by inputting a specific company or board member name.\",\n \"212.A company board data processing system as in claim 211, wherein said searching means is adapted to allow a user to search for a particular board in said board member store by selecting one or more parameters from a pre-defined list.\",\n \"213.A company board data processing system as in claim 1, further comprising: a store storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; wherein said user interface generator is adapted to generate a user interface displaying a list of said pre-defined peer groups; the system further comprising user selector means for receiving a user selection of one or more said pre-defined peer groups from said list; wherein said user interface generator is adapted to generate a user interface displaying at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.\",\n \"214.A company board data processing system as in claim 213, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.\",\n \"215.A company board data processing method comprising: storing parameters related to board members of a plurality of boards organised into boards; and generating a user interface displaying said parameters related to board members for a plurality of boards and/or individual board members to allow comparison of boards, at board member level, and/or board members.\",\n \"216.A company board data processing method as in claim 215, wherein said user interface displays said parameters for each member of at least one board and/or an average value of one or more of said parameters of said members of said at least one board.\",\n \"217.A company board data processing method as in claim 215, further comprising processing parameters for a group of boards or board members to determine at least one of one or more parameter totals, one or more parameter averages and one or more parameter deviations, wherein said user interface displays at least one of said processed parameters for at least one group of boards or board members to allow comparison with at least one board or board member.\",\n \"218.A company board data processing method as in claim 215, wherein said user interface displays at least one of one or more totals, averages and deviations of said parameters related to a group of board members or boards of to allow comparison with at least one board or board member.\",\n \"219.A company Board data processing method as in claim 215, including ranking members of a group of boards and/or board members based on relative value of said parameters; wherein generating a user interface to display said parameters of said boards and/or board members of said group to indicate ranking.\",\n \"220.A company Board data processing method as in claim 217, including ranking groups of boards and/or board members based on relative value of said processed parameters; wherein generating a user interface to display said processed parameters of said groups of boards and/or board members to indicate ranking.\",\n \"221.A company data processing method as in claim 217, wherein said group of boards or board members comprise a user selected and/or pre-defined group of boards or board members assigned as a peer group.\",\n \"222.A company board data processing method as in claim 217, further comprising: receiving a user selection of at least one board and/or board member for inclusion in a peer group; wherein said group of boards or board members comprises a user selected group of boards or board members assigned as a peer group; parameters for constituents of a peer group are processed to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; and said user interface displays at least one of said processed peer group parameters.\",\n \"223.A company board data processing method as in claim 222, further comprising receiving a user selection of at least one board for comparison with at least one board peer group, and said user interface displays the parameters related to members of said at least one selected board, to allow the board to be the subject of analysis.\",\n \"224.A company board data processing method as in claim 222, wherein said user interface displays the parameters related to individual members of said at least one selected board.\",\n \"225.A company board data processing method as in claim 222, further comprising receiving a user selection of at least one board member for comparison with said board member peer group, and said user interface displays the parameters related to said at least one selected board member, to allow said at least one selected board member to be the subject of analysis.\",\n \"226.A company board data processing method as in claim 222, further comprising storing data for one or more user selected peer groups; wherein said user interface displays a list of existing peer groups based on said stored peer group data; a user selection of one or more peer groups is received from said list; and said user interface displays at least one of said processed parameters for at least one of said selected peer groups.\",\n \"227.A company board data processing method as in claim 222, further comprising receiving a user amendment to the constituents of a peer group.\",\n \"228.A company board data processing method as in claim 227, further comprising receiving a user instruction to copy one or more constituents of a peer group to another peer group.\",\n \"229.A company board data processing method of claim 222, further comprising receiving a specific company or board member name as an input from a user, and searching for a particular board in the board member store.\",\n \"230.A company board data processing method as in claim 229, further comprising receiving a user selection of one or more parameters from a pre-defined list, and searching for a particular board in said board member store.\",\n \"231.A company board data processing method as in claim 215, further comprising storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; wherein said user interface displays a list of said pre-defined peer groups; further comprising receiving a user selection of one or more said pre-defined peer groups from said list; and said user interface displays at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.\",\n \"232.A company board data processing method as in claim 231, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.\",\n \"233.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 215.234.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 215.235.A company board data processing system comprising: a board member store storing parameters related to board members of a plurality of boards; user selector means for receiving a user selection of at least one board and/or board member for inclusion in a peer group; a parameter processor for processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; and output means for outputting at least one of the processed peer group parameters.\",\n \"236.A company board data processing system as in claim 235, wherein said user selector means is arranged to receive a user selection of at least one board for comparison with at least one of the board peer groups, and said output means is adapted to output the parameters related to members of said at least one selected board, to allow the or each board to be the subject of analysis.\",\n \"237.A company board data processing system as in claim 235, wherein said output means is adapted to output the parameters related to individual members of the or each selected board.\",\n \"238.A company board data processing system as in claim 235, wherein said user selector means is arranged to receive a user selection of at least one board member for comparison with the board member peer group, and said output means is adapted to output the parameters related to said at least one board member, to allow the or each board members to be the subject of analysis.\",\n \"239.A company board data processing system as in claim 235, further comprising a peer group data store storing data for one or more user selected peer groups; wherein said output means is adapted to output a list of existing peer groups based on said stored peer group data; said user selector means is arranged to receive a user selection of one or more peer groups from said list; and said output means is adapted to output at least one of said processed parameters for the selected peer group.\",\n \"240.A company board data processing system as in claim 235, further comprising a comparison group amendment means allowing a user to amend the constituents of said peer groups.\",\n \"241.A company board data processing system as in claim 240, wherein said peer group amendment means is adapted to allow a user to copy one or more constituents of a peer group to another peer group.\",\n \"242.A company board data processing system as in claim 235, further comprising a searching means arranged to allow a user to search for a particular board and/or board member in said board member store by inputting a specific company or board member name.\",\n \"243.A company board data processing system as in claim 242, wherein said searching means is adapted to allow a user to search for a particular board and/or board member in said board member store by selecting one or more parameters from a pre-defined list.\",\n \"244.A company board data processing system as in claim 235, further comprising: a store storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; wherein said output means is adapted to output a list of said pre-defined peer groups; said user selector means is arranged to receive a user selection of one or more said pre-defined peer groups from said list; and said output means is adapted to output at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.\",\n \"245.A company board data processing system as in claim 244, wherein said stored pre-defined peer group related parameters relate to at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.\",\n \"246.A company board data processing method comprising: storing parameters related to board members of a plurality of boards organised into boards; receiving a user selection of at least one board and/or board member for inclusion in a peer group; processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; and outputting at least one of said processed peer group parameters.\",\n \"247.A company board data processing method as in claim 246, further comprising receiving a user selection of one or more boards for comparison with at least one of said board related peer groups, wherein the parameters related to members of said board are output, to allow the board to be the subject of analysis.\",\n \"248.A company board data processing method as in claim 246, wherein the parameters related to individual members of the or each selected board are output.\",\n \"249.A company board data processing method as in claim 246, further comprising receiving a user selection of one or more board members for comparison with said board member related peer group, wherein the parameters related to said board members are output, to allow the board members to be the subject of analysis.\",\n \"250.A company board data processing method as in any one of claim 246, further comprising storing data for one or more user selected peer groups; outputting a list of existing peer groups based on said stored peer group data; receiving a user selection of one or more peer groups from said list; and outputting at least one of said processed parameters for the selected peer group.\",\n \"251.A company board data processing method as in claim 246, further comprising receiving a user amendment to the constituents of said peer groups.\",\n \"252.A company board data processing method as in claim 251, further comprising receiving a user instruction to copy one or more constituents of a peer group to another peer group.\",\n \"253.A company board data processing method as in claim 246, further comprising receiving a specific company or board member name as an input from a user, and searching for a particular board and/or board member in said board member store.\",\n \"254.A company board data processing method as in claim 253, further comprising receiving a user selection of one or more parameters from a pre-defined list, and searching for a particular board and/or board member in said board member store.\",\n \"255.A company board data processing method as in claim 246, further comprising storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; outputting a list of said pre-defined peer groups; receiving a user selection of one or more said pre-defined peer groups from said list; and outputting at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.\",\n \"256.A company board data processing method as in claim 255, wherein said stored pre-defined peer group related parameters relate to at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.\",\n \"257.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 246.258.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 246.259.A company board data processing system comprising: a store storing pre-defined groups of boards and/or board members comprising peer groups and pre-defined parameters related to said peer groups; output means for outputting a list of said peer groups; and a user selector means for receiving a user selection of one or more of said peer groups from said list; wherein said output means is adapted to output at least one of said pre-defined parameters for the or each selected peer groups.\",\n \"260.A company board data processing system as in claim 259, wherein said pre-defined parameters relate to at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.\",\n \"261.A company board data processing system as in claim 259, further comprising a parameter processor for processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and/or one or more peer group parameter deviations; wherein said output means is adapted to output any of said processed parameters.\",\n \"262.A company board data processing system as in claim 259, wherein said store stores parameters related to board members of a plurality of boards organised into boards; said user selector means is arranged to receive a user selection of one or more said boards for comparison with any said board peer group; and said output means is adapted to output the board member parameters for each member of said board individually, to allow the board to be the subject of analysis.\",\n \"263.A company board data processing system as in claim 259, wherein said user selector means is arranged to receive a user selection of at least one board member for comparison with any said board member peer group, said output means is adapted to output the parameters of said at least one board member, to allow the or each board member to be the subject of analysis.\",\n \"264.A company board data processing system as in claim 259, further comprising a comparison group amendment means allowing a user to amend the constituents of said peer groups.\",\n \"265.A company board data processing system as in claim 264, wherein said peer group amendment means is adapted to allow a user to copy one or more constituents of one said peer group to another said peer group.\",\n \"266.A company board data processing method comprising; storing pre-defined groups of boards and/or board members comprising peer groups and pre-defined parameters related to said peer groups; outputting a list of said peer groups; receiving a user selection of one or more of said peer groups from said list; and outputting at least one of said pre-defined parameters for the or each selected peer groups.\",\n \"267.A company board data processing method as in claim 266, wherein said pre-defined parameters relate to at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.\",\n \"268.A company board data processing method as in claim 266, further comprising processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and/or one or more peer group parameter deviations; wherein any of said processed parameters are output.\",\n \"269.A company board data processing method as in claim 266, further comprising storing parameters related to board members of a plurality of boards organised into boards; receiving a user selection of at least one said board for comparison with any said board peer group; and outputting the board member parameters for each member of said at least one board individually, to allow the or each board to be the subject of analysis.\",\n \"270.A company board data processing method as in claim 266, further comprising receiving a user selection of one or more board members for comparison with any said board member peer group, and outputting the parameters of said board members, to allow the board members to be the subject of analysis.\",\n \"271.A company board data processing method as in claim 266, further comprising receiving a user amendment of the constituents of said peer groups.\",\n \"272.A company board data processing method as in claim 271, further comprising receiving an instruction from a user to copy one or more constituents of one said peer group to another said peer group.\",\n \"273.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 266.274.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 266.275.A company board data processing system comprising: a board member store storing board member related parameters for a plurality of board members; parameter deviation determining means for determining deviations of said board member related parameters from pre-determined parameters; and a user interface generator for generating a user interface to display at least one of said board member related parameters highlighted in accordance with said determined deviations.\",\n \"276.A company board data processing system as in claim 275, wherein said user interface generator is arranged to highlight each said board member related parameter in one of a plurality of colours, wherein said colours relate to the relative value of said determined deviations.\",\n \"277.A company board data processing system as in claim 275, wherein said board member related parameters relate to one or more parameter totals, one or more parameter averages and/or one or more parameter deviations of a plurality of said board members.\",\n \"278.A company board data processing system as in claim 275, wherein said predetermined parameters relate to a group of said board members, comprising a peer group.\",\n \"279.A company board data processing system as in claim 278, wherein said predetermined parameters relate to at least one of one or more totals, one or more averages and one or more deviations of the parameters of the members of said peer group.\",\n \"280.A company board data processing system as in claim 275, further comprising a parameter processor for processing parameters to determine at least one of one or more totals, one or more averages and one or more deviations for said pre-determined parameters; and wherein said user interface generator is adapted to generate said user interface to display said processed parameters.\",\n \"281.A company board data processing system as in claim 275, wherein said parameter deviation determining means is adapted to determine said deviations as a number of standard deviations by which said board member related parameters differ from said pre-determined parameters.\",\n \"282.A company board data processing system as in claim 275, wherein said user interface generator is adapted to generate a user interface to display a table of data, comprising a plurality of cells, each cell containing a said board member related parameter or a said pre-determined parameter, wherein said user interface generator is adapted to highlight said cells containing said board member related parameters in accordance with said determined deviation.\",\n \"283.A company board data processing system as in claim 282, wherein said user interface generator is arranged to highlight each said cell in one of a plurality of colours, wherein said colours relate to the relative value of said determined deviations.\",\n \"284.A company board data processing system as in claim 275, wherein said pre-determined parameters relate to a user selected and/or pre-defined group of board members arranged into a peer group.\",\n \"285.A company board data processing system as in claim 284 further comprising: user selector means for receiving a user selection of at least one board and/or board member for inclusion in a peer group; and a parameter processor for processing parameters for constituents of a peer group to determine said predetermined parameters as at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.\",\n \"286.A company board data processing system as in claim 285, wherein said user selector means is arranged to receive a user selection of one or more boards for comparison with at least one of said board related peer groups, and said user interface generator is adapted to generate the user interface to display the parameters related to members of said board highlighted in accordance with said determined deviation, to allow the board to be the subject of analysis.\",\n \"287.A company board data processing system as in claim 285, wherein said user interface generator is adapted to generate a user interface to display the parameters related to individual members of the or each selected board highlighted in accordance with said determined deviation.\",\n \"288.A company board data processing system as in claim 285, wherein said user selector means is arranged to receive a user selection of at least one board member for comparison with the board member peer group, and said user interface generator is adapted to generate the user interface to display the parameters related to said at least one board member highlighted in accordance with said determined deviations, to allow the or each board member to be the subject of analysis.\",\n \"289.A company board data processing system as in claim 285, further comprising a peer group data store storing data for one or more user selected peer groups; wherein said user interface generator is adapted to generate a user interface to display a list of existing peer groups based on said stored peer group data; said user selector means is arranged to receive a user selection of one or more peer groups from said list; and said user interface generator is adapted to generate the user interface to display at least one of said processed parameters for at least one of said selected peer groups highlighted in accordance with said determined deviations.\",\n \"290.A company board data processing system as in claim 285, further comprising a comparison group amendment means allowing a user to amend the constituents of a peer group.\",\n \"291.A company board data processing system as in claim 290, wherein said peer group amendment means is adapted to allow a user to copy one or more constituents of a peer group to another peer group.\",\n \"292.A company board data processing system as in claim 285, further comprising a searching means arranged to allow a user to search for a particular board in the board member store by inputting a specific company or board member name.\",\n \"293.A company board data processing system as in claim 292, wherein said searching means is adapted to allow a user to search for a particular board in said board member store by selecting one or more parameters from a pre-defined list.\",\n \"294.A company board data processing system as in claim 285, further comprising a store storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; wherein said user interface generator is adapted to generate a user interface to display a list of said pre-defined peer groups; including per group user selector means arranged to receive a user selection of one or more said pre-defined peer groups from said list; wherein said user interface generator is adapted to generate the user interface to display at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.\",\n \"295.A company board data processing system as in claim 294, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.\",\n \"296.A company board data processing method comprising: storing board member related parameters for a plurality of board members; determining deviations of said board member related parameters from pre-determined parameters; and generating a user interface to display at least one of said board member related parameters highlighted in accordance with said determined deviations.\",\n \"297.A company board data processing method as in claim 296, wherein said user interface is generated to highlight each said board member related parameter in one of a plurality of colours, wherein said colours relate to the relative value of said determined deviations.\",\n \"298.A company board data processing method as in claim 296, wherein said board member related parameters relate to one or more parameter totals, one or more parameter averages and/or one or more parameter deviations of a plurality of said board members.\",\n \"299.A company board data processing method as in claim 296, wherein said pre-determined parameters relate to a group of said board members, comprising a peer group.\",\n \"300.A company board data processing method as in claim 299, wherein said pre-determined parameters relate to at least one of one or more totals, one or more averages and one or more deviations of the parameters of the members of said peer group.\",\n \"301.A company board data processing method as in claim 296, further comprising processing parameters to determine at least one of one or more totals, one or more averages and one or more deviations for said pre-determined parameters; and wherein said user interface displays said processed parameters.\",\n \"302.A company board data processing method as in claim 296, wherein said determined deviations comprise a number of standard deviations by which said board member related parameters differ from said pre-determined parameters.\",\n \"303.A company board data processing method as in claim 296, wherein said user interface displays a table of data comprising a plurality of cells, each cell containing a said board member related parameter or a said pre-determined parameter, said cells containing said board member related parameters being highlighted in accordance with said determined deviation.\",\n \"304.A company board data processing method as in claim 303, wherein said user interface highlights each said cell in one of a plurality of colours, wherein said colours relate to the relative value of said determined deviations.\",\n \"305.A company board data processing method as in claim 296 wherein said pre-determined parameters relate to a user selected and/or pre-defined group of board members arranged into a peer group.\",\n \"306.A company board data processing method as in claim 305, further comprising: receiving a user selection of at least one board and/or board member for inclusion in a peer group; and processing parameters for constituents of a peer group to determine said predetermined parameters as at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.\",\n \"307.A company board data processing method as in claim 306, wherein a user selection of one or more boards is received for comparison with at least one of said board related peer groups, and said user interface displays the parameters related to members of said board highlighted in accordance with said determined deviation, to allow the board to be the subject of analysis.\",\n \"308.A company board data processing method as in claim 306, wherein said user interface displays the parameters related to individual members of the or each selected board highlighted in accordance with said determined deviation.\",\n \"309.A company board data processing method as in claim 306, wherein a user selection of at least one board member is received for comparison with the board member peer group, and said user interface displays the parameters related to said at least one board member highlighted in accordance with said determined deviations, to allow the or each board member to be the subject of analysis.\",\n \"310.A company board data processing method as in claim 306, further comprising storing data for one or more user selected peer groups in a peer group data store; wherein said user interface displays a list of existing peer groups based on said stored peer group data; a user selection of one or more peer groups from said list is received; and said user interface displays at least one of said processed parameters for at least one of said selected peer groups highlighted in accordance with said determined deviations.\",\n \"311.A company board data processing method as in claim 306, further comprising receiving a user amendment of the constituents of a peer group.\",\n \"312.A company board data processing method as in claim 311, wherein an instruction is received from a user to copy one or more constituents of a peer group to another peer group.\",\n \"313.A company board data processing method as in claim 306, further comprising receiving a specific company or board member name from a user to search for a particular board in the board member store.\",\n \"314.A company board data processing method as in claim 313, wherein a user selection of one or more parameters from a pre-defined list is received to search for a particular board in said board member store.\",\n \"315.A company board data processing method as in claim 306, further comprising storing pre-defined groups of boards and/or board members comprising peer groups and parameters related to said peer groups; wherein said user interface is generated to display a list of said pre-defined peer groups; a user selection of one or more said pre-defined peer groups from said list is received; and said user interface generates the user interface to display at least one of said stored pre-defined peer group related parameters for said selected pre-defined peer group.\",\n \"316.A company board data processing method as in claim 315, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.\",\n \"317.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 296.318.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 296.319.A company board data processing system comprising: a board member store storing board member related parameters for a plurality of boards; and a user interface generator arranged to generate and display a multi axis plot of one or more of said board member related parameters for one or more boards and/or board members for comparison purposes.\",\n \"320.A company board data processing system as in claim 319, further comprising a processor for determining a scale for at least one axis on said multi axis plot based on at least one of one or more totals, one or more averages and one or more deviations of a plurality of said parameters; wherein said user interface generator is adapted to display at least one axis on said multi axis plot with a scale equal to said determined scale.\",\n \"321.A company board data processing system as in claim 319, wherein said board member store stores said parameters such that said board members are grouped into groups, and said user interface generator is arranged to generate a display of a multi axis plot of at least one of one or more totals, one or more averages or one or more deviations of said parameters of one or more said groups.\",\n \"322.A company board data processing system as in claim 321, wherein said group comprises a board.\",\n \"323.A company board data processing system as in claim 321, further comprising a parameter processor adapted to process parameters for said groups to determine at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations; wherein said user interface generator is adapted to display a plurality of said parameters and/or processed parameters on a multi axis plot, showing each axis for a different parameter type.\",\n \"324.A company board data processing system as in claim 323, wherein said processor is adapted to determine a scale of at least one axis on said multi axis plot based on a predetermined level of deviation of at least one parameter already displayed on that axis from an average of a plurality of said stored and/or said processed parameters for a said group or a plurality of boards and/or board members.\",\n \"325.A company board data processing system as in claim 323, wherein said parameter processor is adapted to determine the scale of at least one parameter axis by determining a minimum axis value of −n standard deviations and a maximum axis value of +m standard deviations of at least one parameter displayed on that axis from an average of a plurality of stored and/or processed parameters related to said group or a plurality of boards and/or board members, where n and m are integers.\",\n \"326.A company board data processing system as in claim 325, wherein said plurality of boards and/or board members comprise all of said boards and/or board members from a specific pre-defined geographic region.\",\n \"327.A company board data processing system as in claim 319 wherein said board member store stores parameters for at least some of said board members as a user selected and/or pre-defined group of board members assigned as a peer group.\",\n \"328.A company board data processing system as in claim 327 further comprising: user selector means for receiving a user selection of at least one board and/or board member for inclusion in a peer group; a parameter processor for processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; and said user interface generator is adapted to generate the user interface to display a multi axis plot of one or more said processed parameters related to said peer group.\",\n \"329.A company board data processing system as in claim 328, wherein said user selector means is arranged to receive a user selection of one or more boards for comparison with at least one of said board peer groups, and said user interface generator is adapted to generate a user interface to display a multi axis plot of one or more parameters related to members of said at least one board, to allow the or each board to be the subject of analysis.\",\n \"330.A company board data processing system as in claim 328, wherein said user interface generator is adapted to generate a user interface to display a multi axis plot of the parameters related to individual members of the or each selected board.\",\n \"331.A company board data processing system as in claim 328, wherein said user selector means is arranged to receive a user selection of at least one board member for comparison with said board member related peer group, and said user interface generator is adapted to generate a user interface to display a multi axis plot of the parameters related to said at least one board member, to allow the or each board member to be the subject of analysis.\",\n \"332.A company board data processing system as in claim 328, wherein said user interface generator is adapted to generate a user interface to display a list of existing peer groups based on stored peer group data in said board member store; said user selector means is arranged to receive a user selection of one or more peer groups from said list; and said user interface generator is adapted to generate a user interface to display a multi axis plot of at least one of said processed parameters for at least one of said selected peer groups.\",\n \"333.A company board data processing system as in claim 328, further comprising a comparison group amendment means allowing a user to amend the constituents of a peer group.\",\n \"334.A company board data processing system as in claim 333, wherein said peer group amendment means is adapted to allow a user to copy one or more constituents of a peer group to another peer group.\",\n \"335.A company board data processing system as in claim 328, further comprising a searching means arranged to allow a user to search for a particular board in the board member store by inputting a specific company or board member name.\",\n \"336.A company board data processing system as in claim 335, wherein said searching means is adapted to allow a user to search for a particular board in said board member store by selecting one or more parameters from a pre-defined list.\",\n \"337.A company board data processing system as in claim 328, wherein said user interface generator is adapted to generate a user interface to display a list of said pre-defined peer groups; said user selector means is arranged to receive a user selection of at least one said pre-defined peer groups from said list; and said user interface generator is adapted to generate a user interface to display a multi axis plot of at least one of said stored pre-defined peer group related parameters for said at least one selected pre-defined peer group.\",\n \"338.A company board data processing system as in claim 337, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.\",\n \"339.A company board data processing method comprising: storing board member related parameters for a plurality of boards; and generating a user interface displaying a multi axis plot of one or more of said board member related parameters for one or more boards and/or board members for comparison purposes.\",\n \"340.A company board data processing method as in claim 339, further comprising determining a scale for at least one axis on said multi axis plot based on at least one of one or more totals, one or more averages and one or more deviations of a plurality of said parameters; wherein said user interface is generated to display at least one axis on said multi axis plot with a scale equal to said determined scale.\",\n \"341.A company board data processing method as in claim 339, wherein said parameters are stored such that said board members are grouped into groups, and said user interface is generated to display a multi axis plot of at least one of one or more totals, one or more averages or one or more deviations of said parameters of one or more said groups.\",\n \"342.A company board data processing method as in claim 341, wherein said group comprises a board.\",\n \"343.A company board data processing method as in claim 341, further comprising processing parameters for said groups to determine at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations; wherein said user interface is generated to display a plurality of said parameters and/or processed parameters on a multi axis plot, showing each axis for a different parameter type.\",\n \"344.A company board data processing method as in claim 343, wherein a scale of at least one axis on said multi axis plot is determined based on a predetermined level of deviation of at least one parameter already displayed on that axis from an average of a plurality of said stored and/or said processed parameters for a said group or a plurality of boards and/or board members.\",\n \"345.A company board data processing method as in claim 343, wherein the scale of at least one parameter axis is determined by determining a minimum axis value of −n standard deviations and a maximum axis value of +m standard deviations of at least one parameter displayed on that axis from an average of a plurality of stored and/or processed parameters related to said group or a plurality of boards and/or board members, where n and m are integers.\",\n \"346.A company board data processing method as in claim 345, wherein said plurality of boards and/or board members comprise all of said boards and/or board members from a specific pre-defined geographic region.\",\n \"347.A company board data processing method as in claim 339 wherein said board member store stores parameters for at least some of said board members as a user selected and/or pre-defined group of board members assigned as a peer group.\",\n \"348.A company board data processing method as in claim 347 further comprising: receiving a user selection of at least one board and/or board member for inclusion in a peer group; and processing parameters for constituents of a peer group to determine at least one of one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations; wherein said user interface generates the user interface to display a multi axis plot of one or more said processed parameters related to said peer group.\",\n \"349.A company board data processing method as in claim 348, wherein a user selection of one or more boards is received for comparison with at least one of said board peer groups, and said user interface is generated to display a multi axis plot of one or more parameters related to members of said at least one board, to allow the or each board to be the subject of analysis.\",\n \"350.A company board data processing method as in claim 348, wherein said user interface is generated to display a multi axis plot of the parameters related to individual members of the or each selected board.\",\n \"351.A company board data processing method as in claim 348, wherein a user selection of at least one board member is received for comparison with said board member related peer group, and said user interface is generated to display a multi axis plot of the parameters related to said at least one board member, to allow the or each board member to be the subject of analysis.\",\n \"352.A company board data processing method as in claim 348, wherein said user interface is generated to display a list of existing peer groups based on stored peer group data in said board member store; the method including receiving a user selection of one or more peer groups from said list; wherein said user interface is generated to display a multi axis plot of at least one of said processed parameters for at least one of said selected peer groups.\",\n \"353.A company board data processing method as in claim 348, further comprising receiving a user amendment of the constituents of a peer group.\",\n \"354.A company board data processing method as in claim 353, wherein an instruction is received from a user to copy one or more constituents of a peer group to another peer group.\",\n \"355.A company board data processing method as in claim 348, further comprising receiving a specific company or board member name from a user to search for a particular board in the board member store.\",\n \"356.A company board data processing method as in claim 355, wherein one or more parameters from a pre-defined list selected by a user is received to search for a particular board in said board member store.\",\n \"357.A company board data processing method as in claim 348, wherein said user interface is generated to display a list of said pre-defined peer groups; a user selection of at least one said pre-defined peer groups from said list is received; and said user interface is generated to display a multi axis plot of at least one of said stored pre-defined peer group related parameters for said at least one selected pre-defined peer group.\",\n \"358.A company board data processing method as in claim 357, wherein said stored pre-defined peer group related parameters relate to at least one of one or more group parameter totals, one or more group parameter averages and one or more group parameter deviations.\",\n \"359.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 339.360.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 339.361.A company board data processing system comprising: access means for accessing a board member store storing parameters related to board members of a plurality of boards organised into boards; and a user interface generator for generating a user interface displaying said parameters related to board members for a plurality of boards to allow comparison of boards, at board member level, and/or board members of boards.\",\n \"362.A company board data processing system as in claim 361, wherein said user interface generator is adapted to generate said user interface to display at least one of one or more totals, averages and deviations of said parameters related to a group of board members or boards to allow comparison with at least one board or board member.\",\n \"363.A company board data processing method comprising: accessing a board member store storing parameters related to board members of a plurality of boards organized into boards; and generating a user interface displaying said parameters related to board members for a plurality of boards to allow comparison of boards, at board member level, and/or board members of boards.\",\n \"364.A company board data processing method as in claim 363, wherein said user interface is generated to display at least one of one or more totals, averages and deviations of said parameters related to a group of board members or boards to allow comparison with at least one board or board member.\",\n \"365.A company board data processing system comprising: a program memory storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 363.366.A carrier medium carrying computer readable instructions for controlling a computer to carry out the method of claim 363.367.A company board data processing system comprising: a store storing a number of shares owned by at least one board member in a company in which they are a board member; means to determine current share price of the company in which said board member is a member of the board; processor to process said number of shares and said share price to determine the value of said shares held by said board member; to determine a wealth index value for at least one said board member, comprising an increase in wealth related to said company of said board member when said share price increases by one percent; and output means for outputting at least one of said determined wealth index values.\",\n \"368.A company board data processing method comprising: storing a number of shares owned by at least one board member in a company in which they are a board member; determining current share price of the company in which said board member is a member of the board; processing the number of shares and the share price to determine the value of said shares held by said board member; determining a wealth index for at least one said board member, comprising an increase in wealth related to said company of said board member when said share price increases by 1%; and outputting at least one of said determined wealth index values.\",\n \"369.A company board data processing system, comprising: a store storing history information for a plurality of board members, said history information including information on previous companies of which said board member has been a board member, and timing information; a graphical display device for generating a graphical display showing performance data for companies together with the history data as a graph showing when the board member was at each company to allow a user to see how the companies have performed in relation to whether or not the board member was a member of the board for the company.\",\n \"370.A system for analysing a created company board, comprising; a store for storing parameters related to a plurality of board members; user selection means for receiving a user selection of at least one board member for inclusion in at least one created company board; a parameter processor to generate board parameters for constituents of said created company board; and output means for outputting said board member parameters and/or said generated board parameters to allow the created board to be the subject of analysis.\",\n \"371.A method of analyzing a created company board, comprising: storing parameters related to a plurality of board members; receiving a user selection of at least one board member for inclusion in at least one created company board; generating board parameters for constituents of said created company board; and outputting said company member parameters and/or said generated board parameters to allow the created board to be the subject of analysis.\",\n \"372.A system for analyzing a created company board, comprising: a program memory storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 371.373.A carrier medium carrying computer readable instructions for controlling a computer to carry out the method of claim 371.374.A board member level recruitment and analysis tool comprising: a store storing parameters related to a plurality of board members; user selection means for receiving a user selection of at least one board member to be the subject of analysis, and a user selection of at least one other board member to be used as a comparison in board member analysis; and a display generator arranged to display the parameters of said at least one user selected board member subject to analysis and said at least one user selected comparison board member to allow recruitment related comparison of the two sets of parameters.\",\n \"375.A board member level recruitment and analysis tool as in claim 374, further comprising: a processor for processing said board member parameters to determine at least one of one or more parameter totals, one or more parameter averages and one or more parameter deviations for a plurality of said user selected board members; wherein said display generator is adapted to display said processed parameters.\",\n \"376.A board member level recruitment and analysis method comprising: storing parameters related to a plurality of board members; receiving a user selection of at least one board member to be the subject of analysis, and a user selection of at least one other board member to be used as a comparison in board member analysis; and displaying the parameters of said at least one user selected board member subject to analysis and said at least one user selected comparison board member to allow recruitment related comparison of the two sets of parameters.\",\n \"377.A board member level recruitment and analysis system, comprising: a program memory storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to: store parameters related to a plurality of board members; receive a user selection of at least one board member to be the subject of analysis, and a user selection of at least one other board member to be used as a comparison in board member analysis; and displaying the parameters of said at least one user selected board member subject to analysis and said at least one user selected comparison board member to allow recruitment related comparison of the two sets of parameters.\",\n \"378.A company board data processing system comprising: a board member store storing board member names for a plurality of board members organized into boards; association determining means for determining associations between said board members; a user interface generator for generating a user interface to display at least one of said board member names and a corresponding list of said determined associations for said board member.\",\n \"379.A company board data processing system as in claim 378, wherein said association determining means is adapted to determine an association to exist between two board members when said two board members both serve on the same two or more independent boards together.\",\n \"380.A company board data processing system as in claim 378, wherein said association determining means is adapted to determine an association to exist between two boards when one board member serves on said two boards at the same time.\",\n \"381.A company board data processing system as in claim 378, wherein said association determining means is adapted to determine indirect associations between two or more boards and/or board members via a plurality of other associations; and said user interface generator is adapted to generate a user interface to display at least one board and/or board member and a list of said network of determined associations to at least one other board and/or board member.\",\n \"382.A company board data processing system as in claim 378, wherein said user interface generator is adapted to generate a user interface to display a list of board members and/or boards based on said stored board member data; further comprising a user selector means for receiving a user selection of two or more boards and/or board members from said list; wherein said user interface generator is adapted to generate a user interface to display said names of said boards and/or board members and a list of said determined associations between said boards and/or board members.\",\n \"383.A company board data processing system as in claim 381, wherein said user interface generator is adapted to generate a user interface to display a list of board members and/or boards based on said stored board member data; further comprising a user selector means for receiving a user selection of two or more boards and/or board members from said list; wherein said user interface generator is adapted to generate a user interface to display said names of said boards and/or board members and a list of said determined indirect associations between said boards and/or board members.\",\n \"384.A company board data processing method comprising: storing board member names for a plurality of board members organized into boards; determining associations between said board members; generating a user interface to display at least one of said board member names and a corresponding list of said determined associations for said board member.\",\n \"385.A company board data processing method as in claim 384, wherein said determining of associations includes determining an association to exist between two board members when said two board members both serve on the same two or more independent boards together.\",\n \"386.A company board data processing method as in claim 384, wherein said determining of associations includes determining an association to exist between two boards when one board member serves on said two boards at the same time.\",\n \"387.A company board data processing method as in claim 384 further comprising; determining indirect associations between two or more boards and/or board members via a plurality of other associations; wherein said user interface displays at least one board and/or board member and a list of said network of determined associations to at least one other board and/or board member.\",\n \"388.A company board data processing method as in claim 384, wherein said user interface displays a list of board members and/or boards based on said stored board member data; further comprising receiving a user selection of two or more boards and/or board members from said list; and in response to said user selection said user interface displays said names of said selected boards and/or board members and a list of said determined associations between said boards and/or board members.\",\n \"389.A company board data processing method as in claim 387, wherein said user interface displays a list of board members and/or boards based on said stored board member data; further comprising receiving a user selection of two or more boards and/or board members from said list; and in response to said user selection said user interface displays said names of said selected boards and/or board members and a list of said determined indirect associations between said boards and/or board members.\",\n \"390.A company board data processing system comprising a program member storing processor implementable instructions; and a processor for reading and implementing the instructions stored in the program memory; wherein the instructions stored in the program memory comprise instructions for controlling the processor to implement the method of claim 378.391.A carrier medium carrying computer readable instructions for controlling the computer to carry out the method of claim 378.392.A method of fund management using the system of claim 1.393.A method of fund management using the system of claim 235.394.A method of fund management using the system of claim 259.395.A method of fund management using the system of claim 275.396.A method of fund management using the system of claim 319.397.A method of fund management using the system of claim 361.398.A method of fund management using the system of claim 367.399.A method of fund management using the system of claim 369.400.A method of fund management using the system of claim 370.401.A method of fund management using the system of claim 374.402.A method of fund management using the system of claim 375.403.A method of fund management using the system of claim 378.404.A method of recruiting board members using the system of claim 1.405.A method of recruiting board members using the system of claim 235.406.A method of recruiting board members using the system of claim 259.407.A method of recruiting board members using the system of claim 275.408.A method of recruiting board members using the system of claim 319.409.A method of recruiting board members using the system of claim 361.410.A method of recruiting board members using the system of claim 367.411.A method of recruiting board members using the system of claim 369.412.A method of recruiting board members using the system of claim 370.413.A method of recruiting board members using the system of claim 374.414.A method of recruiting board members using the system of claim 375.415.A method of recruiting board members using the system of claim 378.\"\n ],\n [\n \"<SOH> BACKGROUND <EOH>Investing on the Stock Markets inevitably involves a risk on behalf of the investor, equities can go up or down and there is never any guarantee of a positive return from your investment.\",\n \"However this uncertainty can be reduced considerably by investing wisely and more importantly by investing only in things that you have knowledge of.\",\n \"You will have more chance of winning if you bet on a horse based on its form than you will if you take a guess from a list of starters without any knowledge of any of them, and so it is with the equity markets that a sensible investor will only buy or sell a stock based on sound information.\",\n \"It is the requirement for this information across the stock markets of the world which has led to the existence of the world-wide Investment Banks and a plethora of financial advisors.\",\n \"It is almost impossible for a fund manager or private investor to be able to make an informed decision on a stock without some form of advice on the economic conditions of that company, its past history, future prospects, products, staff, etc.\",\n \".\",\n \".\",\n \".\",\n \"This advice can come in many forms depending on the information sought, for instance there are a large number of sources of company focussed economic data, information such as the price/earnings ratio, the profit & loss figures etc .\",\n \".\",\n \".\",\n \", well known financial statistics which can give an indication as to the stock's current worth and potential.\",\n \"However, there are other forms of information that are more subjective by their very nature, and many of the large investment houses have teams of analysts trying to get behind the financial figures to tell the real story.\",\n \"The problem with such information is that it is not as reliable as the hard economic facts, it often emanates from person to person meetings, press conferences and rumour.\",\n \"It is not possible for a fund manager or investor to attend every annual general meeting, read every end of year report and interview every Board of Directors, and hence it is the analyst's job to try to do this for the investor, and it is left to the investor to attempt to read the analyst's report and pick out the information that he is looking for.\",\n \"This, inevitably increases the risk of following the information and advice supplied, especially as two different analysts will often have a completely opposite view on the potential of any particular company.\",\n \"Unfortunately there are very few alternatives, and whilst the use of analyst reports helps it still rarely allows for comparison across the whole spectrum of the market because analysts will often have a specialised sector and hence comparative data across a range of stocks in this field of information can be difficult to find and often lacking in user friendliness.\",\n \"Hence it is the object of the proposed invention to supply a user friendly tool to aid in information provision.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>A first aspect of the present invention provides a company board data processing system and method which provides a user interface platform to supply information related to members of boards of companies to enable comparison of board members or boards at board member level.\",\n \"Parameters related to board members of a plurality of boards are stored.\",\n \"The boards of which the board members are members is indicated in the stored parameters.\",\n \"A user interface is generated which comprises a display of the parameters related to board members for a plurality of boards and/or individual board members to, allow comparison of board members and/or boards at board member level.\",\n \"Thus this aspect of the present invention provides a tool which can be used by analysts to review parameters related to boards at the level of granularity of board member parameters.\",\n \"In one embodiment of this aspect of the present invention, parameters for a group of board members such as a board or some other predetermined group is processed to determine process parameters which can comprise one or a number of: parameter totals, parameter averages and parameter deviations.\",\n \"Thus the user interface can display such process parameters for comparison with boards at board member level or with individual board members.\",\n \"In one embodiment of this aspect of the present invention the group can comprise a group of boards in which case the process parameters comprise parameters processed for a number of boards and thus represents totals, averages or deviations for the boards.\",\n \"This enables the board as a whole and individual members of the board to be compared to a number of boards.\",\n \"In an alternative embodiment of this aspect of the present invention the group comprises a group of board members.\",\n \"The group of board members can comprise board members of a single board, or board members across a number of boards.\",\n \"This enables individual board members to be compared with a peer group of board members from a number of boards.\",\n \"A second aspect of the present invention provides a company board data processing system and method in which parameters related to board members of a plurality of boards is stored.\",\n \"The system receives a user selection of at least one board and/or board member for inclusion in a peer group.\",\n \"Parameters for constituents of a peer group are processed to determine processed parameters for the peer group and these processed parameters are output.\",\n \"Preferably the processing determines one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.\",\n \"This aspect of the present invention provides a useful tool to enable a user to select and form a peer group comprised of a number of board members which need not be in the same board.\",\n \"This aspect of the present invention also allows the selection of boards for inclusion in a board peer group.\",\n \"Parameters for boards are processed to determine process parameters for the board peer group.\",\n \"Thus a board peer group can be used for comparison with a board to allow comparative analysis.\",\n \"In one embodiment of this aspect of the present invention the peer group data is stored for at least one user selected peer group.\",\n \"The user interface can display a list of existing peer groups to allow a user to select one or more of the existing peer groups.\",\n \"Process parameters for the or each selected peer group can then be output for display, e.g.\",\n \"for comparative purposes.\",\n \"In one embodiment of this aspect of the present invention, the constituents of the peer groups can be amended.\",\n \"This amendment can take the form of allowing constituents of a peer group, e.g.\",\n \"a board member or board, to be transferred from one peer group to another.\",\n \"In another embodiment of this aspect of the present invention predefined groups of boards and/or board members are stored with parameters related to the peer groups as predefined peer groups.\",\n \"The user interface can display a list of the predefined peer groups to allow a user to select one or more of them for output for comparative purposes.\",\n \"Another aspect of the present invention provides a company board data processing system in which predefined groups of boards and/or board members comprising peer groups and predefined parameters related to the peer groups are stored.\",\n \"The system is arranged to output a list of the peer groups to allow a user to select one or more of the peer groups from the list.\",\n \"As a result of the selection at least one of the predefined parameters for the or each selected peer group is output.\",\n \"The parameters can be displayed and used for comparative purposes.\",\n \"In this aspect of the present invention a peer group which can comprise parameters for groups of boards and/or board members is predefined.\",\n \"In one embodiment, however, a user can modify a predefined peer group by adding constituents, i.e.\",\n \"boards or board members, or removing constituents.\",\n \"Also, constituents can be moved from group to group.\",\n \"Thus this embodiment provides a flexible tool for comparison of board members and boards with peer groups.\",\n \"In one embodiment the predefined parameters for the peer groups comprise process parameters.\",\n \"The process parameters can comprise peer group parameter totals, peer group parameter averages and peer group parameter deviations.\",\n \"Another aspect of the present invention provides a company board data processing system and method in which board member related parameters for a plurality of board members is stored.\",\n \"A deviation of said board member related parameters from predetermined parameters is determined and this is used by a user interface to generate a display of at least one board member related parameter highlighted in accordance with the determined deviations.\",\n \"This embodiment of the present invention provides a system in which deviations of parameters related to board members from predetermined parameters is displayed in a clear form by highlighting such parameters in accordance with the deviations.\",\n \"This allows these deviations and patterns in these deviations to be easily distinguishable.\",\n \"In one embodiment of the present invention the user interface is generated to highlight each board member related parameter in one of a plurality of colours, where the colour relates to the relative value of the determined deviation.\",\n \"In one embodiment of this aspect of the present invention the predetermined parameters relate to a group of board members comprising a peer group.\",\n \"In this embodiment predetermined parameters can relate to totals, averages and/or deviations of the parameters of the members of the peer group.\",\n \"In one embodiment of this aspect of the present invention the user interface generates a table of data comprising a plurality of cells.\",\n \"Each cell contains a board member related parameter or a predetermined parameter.\",\n \"Cells containing the board member related parameters are highlighted in accordance with the determined deviation.\",\n \"In a preferred embodiment these cells are highlighted in one of a plurality of colours, wherein the colours relate to the relative value of the determined deviations.\",\n \"In another aspect of this aspect of the present invention there is provided a company board data processing system and method in which board member related parameters are stored for a plurality of boards.\",\n \"A user interface generates a multi-axis plot of one or more of the board member related parameters for one or more boards and/or board members to allow comparison of these parameters.\",\n \"Thus this aspect of the present invention provides a graphical user interface tool for the visualization of company board data at a company board member level of granularity.\",\n \"In one embodiment of this aspect of the present invention the board members can be grouped into groups and the user interface can generate a display of a multi-axis plot of parameters related to a group of board members.\",\n \"In one embodiment this can be used for comparative purposes to allow parameters related to a board or individual members of a board to be plotted against the group.\",\n \"In one embodiment of the present invention the axes of the multi-axis plot can be scaled based on a plurality of parameters for a plurality of board members.\",\n \"This can comprise totals, averages or deviations for parameters for a plurality of board members.\",\n \"A further aspect of the present invention provides a company board data processing system in which a number of shares owned by at least one board member in a company in which they are a board member are stored.\",\n \"The current share price of the company in which the board member is a member of the board is determined and used with the number of shares to determine the value of the shares held by the board member to determine a wealth index for the board member.\",\n \"The wealth index comprises an increase in wealth related to the company when the share price increases by an amount, e.g.\",\n \"1%.\",\n \"This aspect of the present invention provides an indicator of the wealth related motivation of a board member to improve the performance of a company.\",\n \"If a board member holds a large number of shares, they have a personal interest in improving the performance of a company so that the share price increases.\",\n \"Another aspect of the present invention provides a system for analyzing a created company board.\",\n \"Parameters related to a plurality of board members are stored.\",\n \"A user can select at least one board member for inclusion in at least one created company board.\",\n \"Board parameters for constituents of the created company board are generated.\",\n \"The board member parameters and/or the generated board parameters can be output to allow the created board to be the subject of analysis.\",\n \"Thus this aspect of the present invention provides a means by which a user of the system can create a “fantasy” board comprised of a number of board members currently members of other boards.\",\n \"Parameters related to the board members for the created company board can then be analysed.\",\n \"Thus this aspect of the present invention can be used as a tool by which users can identify suitable board members for building a company board for a new company or for replacing board members of an existing company board.\",\n \"A further aspect of the present invention provides a board member level recruitment and analysis tool and method in which parameters related to a plurality of board members are stored.\",\n \"A user is able to select at least one board member to be the subject of analysis and to select at least one other board member to be used as a comparison in the board member analysis.\",\n \"A user interface generates a display of the parameters of the selected board members subject to analysis and the selected comparison board members to allow recruitment related comparison of the two sets of parameters.\",\n \"Another aspect of the present invention provides a company board data processing system and method in which a graphical display is generated to show the performance of a company and a board member history on a time basis.\",\n \"This aspect of the present invention allows a user to identify the effect of a board member on the performance of a company.\",\n \"Although the present invention has been described hereinabove with reference to a number of separate aspects, in accordance with the present invention, any aspect of the present invention described hereinabove can be used in conjunction with any other aspect of the present invention.\",\n \"The present invention is applicable to financial analysis and in management of companies.\",\n \"For example, the present invention can be used by fund managers for managing a portfolio of investments by analyzing company board member parameters.\",\n \"The presentation of the data can be by any suitable output means such as a display or printer.\",\n \"The output can also be an output over a network to provide for presentation, e.g.\",\n \"display, at a remote location.\",\n \"The present invention is applicable to a stand alone system or a network system in which the server receives requests from a user of a client and outputs data for presentation to the user by the client.\",\n \"All aspects of the present invention can be implemented on dedicated hardware, a combination of dedicated hardware and suitably programmed hardware, or on a programmed general purpose computer.\",\n \"The present invention thus encompasses computer readable instructions for controlling a computer to carry out the methods.\",\n \"Computer readable instructions can be provided to a general purpose computer on any suitable carrier medium such as a transient medium, e.g.\",\n \"an electrical, optical, microwave, acoustic or radio frequency signal (e.g.\",\n \"a signal (carrier wave) carrying computer readable instructions over a network such as the Internet), or a storage medium such as a floppy disk, hard disk, CD-ROM or solid state memory device.\"\n ],\n [\n \"FIELD OF INVENTION This invention relates to a data processing system and method for processing and presenting a wide range of data related to the Board of Directors of companies, as well as the members of those Boards individually.\",\n \"BACKGROUND Investing on the Stock Markets inevitably involves a risk on behalf of the investor, equities can go up or down and there is never any guarantee of a positive return from your investment.\",\n \"However this uncertainty can be reduced considerably by investing wisely and more importantly by investing only in things that you have knowledge of.\",\n \"You will have more chance of winning if you bet on a horse based on its form than you will if you take a guess from a list of starters without any knowledge of any of them, and so it is with the equity markets that a sensible investor will only buy or sell a stock based on sound information.\",\n \"It is the requirement for this information across the stock markets of the world which has led to the existence of the world-wide Investment Banks and a plethora of financial advisors.\",\n \"It is almost impossible for a fund manager or private investor to be able to make an informed decision on a stock without some form of advice on the economic conditions of that company, its past history, future prospects, products, staff, etc.\",\n \".\",\n \".\",\n \".\",\n \"This advice can come in many forms depending on the information sought, for instance there are a large number of sources of company focussed economic data, information such as the price/earnings ratio, the profit & loss figures etc .\",\n \".\",\n \".\",\n \", well known financial statistics which can give an indication as to the stock's current worth and potential.\",\n \"However, there are other forms of information that are more subjective by their very nature, and many of the large investment houses have teams of analysts trying to get behind the financial figures to tell the real story.\",\n \"The problem with such information is that it is not as reliable as the hard economic facts, it often emanates from person to person meetings, press conferences and rumour.\",\n \"It is not possible for a fund manager or investor to attend every annual general meeting, read every end of year report and interview every Board of Directors, and hence it is the analyst's job to try to do this for the investor, and it is left to the investor to attempt to read the analyst's report and pick out the information that he is looking for.\",\n \"This, inevitably increases the risk of following the information and advice supplied, especially as two different analysts will often have a completely opposite view on the potential of any particular company.\",\n \"Unfortunately there are very few alternatives, and whilst the use of analyst reports helps it still rarely allows for comparison across the whole spectrum of the market because analysts will often have a specialised sector and hence comparative data across a range of stocks in this field of information can be difficult to find and often lacking in user friendliness.\",\n \"Hence it is the object of the proposed invention to supply a user friendly tool to aid in information provision.\",\n \"SUMMARY OF THE INVENTION A first aspect of the present invention provides a company board data processing system and method which provides a user interface platform to supply information related to members of boards of companies to enable comparison of board members or boards at board member level.\",\n \"Parameters related to board members of a plurality of boards are stored.\",\n \"The boards of which the board members are members is indicated in the stored parameters.\",\n \"A user interface is generated which comprises a display of the parameters related to board members for a plurality of boards and/or individual board members to, allow comparison of board members and/or boards at board member level.\",\n \"Thus this aspect of the present invention provides a tool which can be used by analysts to review parameters related to boards at the level of granularity of board member parameters.\",\n \"In one embodiment of this aspect of the present invention, parameters for a group of board members such as a board or some other predetermined group is processed to determine process parameters which can comprise one or a number of: parameter totals, parameter averages and parameter deviations.\",\n \"Thus the user interface can display such process parameters for comparison with boards at board member level or with individual board members.\",\n \"In one embodiment of this aspect of the present invention the group can comprise a group of boards in which case the process parameters comprise parameters processed for a number of boards and thus represents totals, averages or deviations for the boards.\",\n \"This enables the board as a whole and individual members of the board to be compared to a number of boards.\",\n \"In an alternative embodiment of this aspect of the present invention the group comprises a group of board members.\",\n \"The group of board members can comprise board members of a single board, or board members across a number of boards.\",\n \"This enables individual board members to be compared with a peer group of board members from a number of boards.\",\n \"A second aspect of the present invention provides a company board data processing system and method in which parameters related to board members of a plurality of boards is stored.\",\n \"The system receives a user selection of at least one board and/or board member for inclusion in a peer group.\",\n \"Parameters for constituents of a peer group are processed to determine processed parameters for the peer group and these processed parameters are output.\",\n \"Preferably the processing determines one or more peer group parameter totals, one or more peer group parameter averages and one or more peer group parameter deviations.\",\n \"This aspect of the present invention provides a useful tool to enable a user to select and form a peer group comprised of a number of board members which need not be in the same board.\",\n \"This aspect of the present invention also allows the selection of boards for inclusion in a board peer group.\",\n \"Parameters for boards are processed to determine process parameters for the board peer group.\",\n \"Thus a board peer group can be used for comparison with a board to allow comparative analysis.\",\n \"In one embodiment of this aspect of the present invention the peer group data is stored for at least one user selected peer group.\",\n \"The user interface can display a list of existing peer groups to allow a user to select one or more of the existing peer groups.\",\n \"Process parameters for the or each selected peer group can then be output for display, e.g.\",\n \"for comparative purposes.\",\n \"In one embodiment of this aspect of the present invention, the constituents of the peer groups can be amended.\",\n \"This amendment can take the form of allowing constituents of a peer group, e.g.\",\n \"a board member or board, to be transferred from one peer group to another.\",\n \"In another embodiment of this aspect of the present invention predefined groups of boards and/or board members are stored with parameters related to the peer groups as predefined peer groups.\",\n \"The user interface can display a list of the predefined peer groups to allow a user to select one or more of them for output for comparative purposes.\",\n \"Another aspect of the present invention provides a company board data processing system in which predefined groups of boards and/or board members comprising peer groups and predefined parameters related to the peer groups are stored.\",\n \"The system is arranged to output a list of the peer groups to allow a user to select one or more of the peer groups from the list.\",\n \"As a result of the selection at least one of the predefined parameters for the or each selected peer group is output.\",\n \"The parameters can be displayed and used for comparative purposes.\",\n \"In this aspect of the present invention a peer group which can comprise parameters for groups of boards and/or board members is predefined.\",\n \"In one embodiment, however, a user can modify a predefined peer group by adding constituents, i.e.\",\n \"boards or board members, or removing constituents.\",\n \"Also, constituents can be moved from group to group.\",\n \"Thus this embodiment provides a flexible tool for comparison of board members and boards with peer groups.\",\n \"In one embodiment the predefined parameters for the peer groups comprise process parameters.\",\n \"The process parameters can comprise peer group parameter totals, peer group parameter averages and peer group parameter deviations.\",\n \"Another aspect of the present invention provides a company board data processing system and method in which board member related parameters for a plurality of board members is stored.\",\n \"A deviation of said board member related parameters from predetermined parameters is determined and this is used by a user interface to generate a display of at least one board member related parameter highlighted in accordance with the determined deviations.\",\n \"This embodiment of the present invention provides a system in which deviations of parameters related to board members from predetermined parameters is displayed in a clear form by highlighting such parameters in accordance with the deviations.\",\n \"This allows these deviations and patterns in these deviations to be easily distinguishable.\",\n \"In one embodiment of the present invention the user interface is generated to highlight each board member related parameter in one of a plurality of colours, where the colour relates to the relative value of the determined deviation.\",\n \"In one embodiment of this aspect of the present invention the predetermined parameters relate to a group of board members comprising a peer group.\",\n \"In this embodiment predetermined parameters can relate to totals, averages and/or deviations of the parameters of the members of the peer group.\",\n \"In one embodiment of this aspect of the present invention the user interface generates a table of data comprising a plurality of cells.\",\n \"Each cell contains a board member related parameter or a predetermined parameter.\",\n \"Cells containing the board member related parameters are highlighted in accordance with the determined deviation.\",\n \"In a preferred embodiment these cells are highlighted in one of a plurality of colours, wherein the colours relate to the relative value of the determined deviations.\",\n \"In another aspect of this aspect of the present invention there is provided a company board data processing system and method in which board member related parameters are stored for a plurality of boards.\",\n \"A user interface generates a multi-axis plot of one or more of the board member related parameters for one or more boards and/or board members to allow comparison of these parameters.\",\n \"Thus this aspect of the present invention provides a graphical user interface tool for the visualization of company board data at a company board member level of granularity.\",\n \"In one embodiment of this aspect of the present invention the board members can be grouped into groups and the user interface can generate a display of a multi-axis plot of parameters related to a group of board members.\",\n \"In one embodiment this can be used for comparative purposes to allow parameters related to a board or individual members of a board to be plotted against the group.\",\n \"In one embodiment of the present invention the axes of the multi-axis plot can be scaled based on a plurality of parameters for a plurality of board members.\",\n \"This can comprise totals, averages or deviations for parameters for a plurality of board members.\",\n \"A further aspect of the present invention provides a company board data processing system in which a number of shares owned by at least one board member in a company in which they are a board member are stored.\",\n \"The current share price of the company in which the board member is a member of the board is determined and used with the number of shares to determine the value of the shares held by the board member to determine a wealth index for the board member.\",\n \"The wealth index comprises an increase in wealth related to the company when the share price increases by an amount, e.g.\",\n \"1%.\",\n \"This aspect of the present invention provides an indicator of the wealth related motivation of a board member to improve the performance of a company.\",\n \"If a board member holds a large number of shares, they have a personal interest in improving the performance of a company so that the share price increases.\",\n \"Another aspect of the present invention provides a system for analyzing a created company board.\",\n \"Parameters related to a plurality of board members are stored.\",\n \"A user can select at least one board member for inclusion in at least one created company board.\",\n \"Board parameters for constituents of the created company board are generated.\",\n \"The board member parameters and/or the generated board parameters can be output to allow the created board to be the subject of analysis.\",\n \"Thus this aspect of the present invention provides a means by which a user of the system can create a “fantasy” board comprised of a number of board members currently members of other boards.\",\n \"Parameters related to the board members for the created company board can then be analysed.\",\n \"Thus this aspect of the present invention can be used as a tool by which users can identify suitable board members for building a company board for a new company or for replacing board members of an existing company board.\",\n \"A further aspect of the present invention provides a board member level recruitment and analysis tool and method in which parameters related to a plurality of board members are stored.\",\n \"A user is able to select at least one board member to be the subject of analysis and to select at least one other board member to be used as a comparison in the board member analysis.\",\n \"A user interface generates a display of the parameters of the selected board members subject to analysis and the selected comparison board members to allow recruitment related comparison of the two sets of parameters.\",\n \"Another aspect of the present invention provides a company board data processing system and method in which a graphical display is generated to show the performance of a company and a board member history on a time basis.\",\n \"This aspect of the present invention allows a user to identify the effect of a board member on the performance of a company.\",\n \"Although the present invention has been described hereinabove with reference to a number of separate aspects, in accordance with the present invention, any aspect of the present invention described hereinabove can be used in conjunction with any other aspect of the present invention.\",\n \"The present invention is applicable to financial analysis and in management of companies.\",\n \"For example, the present invention can be used by fund managers for managing a portfolio of investments by analyzing company board member parameters.\",\n \"The presentation of the data can be by any suitable output means such as a display or printer.\",\n \"The output can also be an output over a network to provide for presentation, e.g.\",\n \"display, at a remote location.\",\n \"The present invention is applicable to a stand alone system or a network system in which the server receives requests from a user of a client and outputs data for presentation to the user by the client.\",\n \"All aspects of the present invention can be implemented on dedicated hardware, a combination of dedicated hardware and suitably programmed hardware, or on a programmed general purpose computer.\",\n \"The present invention thus encompasses computer readable instructions for controlling a computer to carry out the methods.\",\n \"Computer readable instructions can be provided to a general purpose computer on any suitable carrier medium such as a transient medium, e.g.\",\n \"an electrical, optical, microwave, acoustic or radio frequency signal (e.g.\",\n \"a signal (carrier wave) carrying computer readable instructions over a network such as the Internet), or a storage medium such as a floppy disk, hard disk, CD-ROM or solid state memory device.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS Embodiments of the present invention will now be described with reference to the accompanying drawings, in which: FIG.\",\n \"1 is a schematic diagram of a company board data processing system in accordance with an embodiment of the present invention; FIG.\",\n \"2 illustrates the top level structure of the user interface information pages; FIG.\",\n \"3 illustrates the information page structure of the Board Associations data section; FIG.\",\n \"4 illustrates the information page structure of the Board Remuneration data section; FIG.\",\n \"5 illustrates the information page structure of the Board Characteristics data section; FIG.\",\n \"6 illustrates the information page structure of the Director Associations data section; FIG.\",\n \"7 illustrates the information page structure of the Director Remuneration data section; FIG.\",\n \"8 illustrates the information page structure of the Director Characteristics data section; FIG.\",\n \"9 is a screen shot of a user interface ‘Board Summary’ information page; FIG.\",\n \"10 is a screen shot of a user interface ‘Board Single & Multi Associations’ information page; FIG.\",\n \"11 is a screen shot of a user interface ‘Board Remuneration Summary Table’ information page; FIG.\",\n \"12 is a screen shot of a user interface ‘Board Remuneration Summary Bubble Chart’ information page; FIG.\",\n \"13 is a screen shot of a user interface ‘Board Remuneration Ratios’ information page; FIG.\",\n \"14 is a screen shot of a user interface ‘Board Characteristics Network/Biographical’ information page; FIG.\",\n \"15 is a screen shot of a user interface ‘Board Characteristics Change Table’ information page; FIG.\",\n \"16 is a screen shot of a user interface ‘Board Characteristics Diversity Table’ information page; FIG.\",\n \"17 is a screen shot of a user interface ‘Board Characteristics Diversity Radar Chart information page; FIG.\",\n \"18 is a screen shot of a user interface ‘Board Peer Group Management’ information page; FIG.\",\n \"19 is a screen shot of a user interface ‘Board Peer Group Member’ information page; FIG.\",\n \"20 is a screen shot of a user interface ‘Board Advanced Search’ information page, FIG.\",\n \"21 is a screen shot of a user interface ‘Board Search Results’ information page; FIG.\",\n \"22 is a screen shot of a user interface ‘Director Summary’ information page; FIG.\",\n \"23 is a screen shot of a user interface ‘Director Single & Multi Associations’ information page; FIG.\",\n \"24 is a screen shot of a user interface ‘Director Remuneration Ratios’ information page; FIG.\",\n \"25 is a screen shot of a user interface ‘Director Characteristics Roles’ information page; FIG.\",\n \"26 is a screen shot of a user interface ‘Director Characteristics Experience Table’ information page; FIG.\",\n \"27 is a screen shot of a user interface ‘Director Characteristics Experience Radar Chart’ information page; FIG.\",\n \"28 is a screen shot of a user interface ‘Director Peer Group Management’ information page; FIG.\",\n \"29 is a screen shot of a user interface ‘Director Peer Group Member’ information page; FIG.\",\n \"30 is a screen shot of a user interface ‘Director Advanced Search’ information page; FIG.\",\n \"31 is a screen shot of a user interface ‘Director Search Results’ information page; FIG.\",\n \"32 is a screen shot of a user interface ‘Board Timeline’ information page; FIG.\",\n \"33 is a screen shot of a user interface ‘Board Share Price History’ information page; FIG.\",\n \"34 is a screen shot of a user interface ‘Board and Shareholder Returns’ information page; FIG.\",\n \"35 is a screen shot of a user interface ‘Board Time Analysis’ information page; FIG.\",\n \"36 is a screen shot of a user interface ‘Board Experience over Time’ information page; FIG.\",\n \"37 is a screen shot of a user interface ‘Director Multi Associations’ information page; FIG.\",\n \"38 is a screen shot of a user interface ‘Director Share Price History’ information page; FIG.\",\n \"39 is a screen shot of a user interface ‘Board Peer Group Alerts Set-Up’ information page; FIG.\",\n \"40 is a screen shot of a user interface ‘Board Peer Group Alerts Results’ information page; FIG.\",\n \"41 is a screen shot of a user interface ‘Director Peer Group Alerts Set-Up’ information page; and FIG.\",\n \"42 is a screen shot of a user interface ‘Director Peer Group Alerts Results’ information page.\",\n \"DESCRIPTION OF PREFERRED EMBODIMENTS General Introduction There are many systems that provide information regarding companies listed on the stock markets of the world, but that there is a gap in the market for an analysis tool that is focussed on the actual members of the Boards of these companies.\",\n \"Such information is currently hard to find and more importantly difficult to analyse and often unreliable.\",\n \"The system of the embodiment of the present invention allows people to view and analyse the people who actually run these listed companies and hence offer a platform for people to form an indicative view of another very important aspect in measuring the potential success, or otherwise, of such companies that has so far been very difficult to grasp.\",\n \"The system offered by the embodiment of the current invention allows the user to select an entire Board and analyse and compare the credentials of each member of that Board.\",\n \"Also, one or more other Boards can be selected, by the user, and corresponding data can be displayed simultaneously to allow a comparison of the selected Boards against one another, hence providing the statistics that can be used to measure the relative merits of one Board against another.\",\n \"This system provides a platform of user interactive analysis that has not been previously available and that will greatly benefit anyone investing in stock market listed companies.\",\n \"However, the usefulness of such a tool is not restricted to the financial markets.\",\n \"Anyone who wished to compare the skills, experience and background history of one or more Director level individuals, say for recruitment purposes, would also find this tool very effective in their analysis.\",\n \"The system allows the user to select an individual and view data on all of his Board-level roles, and also offers the chance to select one or more other individuals to be displayed simultaneously and used in the analysis.\",\n \"Hardware The preferred embodiment of the current invention is illustrated in FIG.\",\n \"1 and comprises an architecture structure that is made up of a Data Repository System, 1, a Database System, 2, 3, 4, 5, a Data Processor, 6, a web Server system, 7, 8, 9, and a User's computer, 11.The Data Repository System, 1, is located at a central data entry and management location, 100.It comprises a unified data entry point and is the tool with which all raw data is entered into the system.\",\n \"The Data Repository System 1 acts as a master database for the data in the system and receives data via one of two methods: manual data entry; or automatic data entry.\",\n \"The manual data entry system comprises a computer system with a user interface based at the central data entry and management location, 100.This user interface allows manual entry of new data raw data parameters as well as the entry of some manual analytics.\",\n \"It comprises two interfaces, one for the entry of data relating to a Director Profile and the other for the entry of data relating to a Board profile.\",\n \"The raw data comprises simple numerical values, such as a Director's salary, that are described hereafter.\",\n \"The manual analytics data entry comprises such data as, for example, a company time-line share price graph that has been manually amended to display key corporate actions during the time period displayed, for example the departure of a CEO from the Board or a new company share issue, such as that shown in FIG.\",\n \"33.The automatic data entry system comprises on on-line feed to a number of financial data providers, such as FT, AFX and Hernscott.\",\n \"This data is downloaded periodically, daily in the case of the preferred embodiment of the invention, into a data store, where it is checked both by manual and automatic means before being populated onto the Master Database.\",\n \"The raw data parameters that are entered via the manual and automatic data entry systems comprise parameters related to Board members and parameters related to Boards and include: Salary Bonus Pension Contributions Other remuneration Role Number of Options awarded Stock Price Option Exercise Price Options time to expiry Dividend Yield Number of Directors at the start of the Year Number of Directors at the end of the Year Equity awarded LTIP Contributions awarded Number of Executives and Non-executives in the company Age of Director Time in Company of Director Time on Board of Director Board's policy retirement Age Time to retirement for Director Name Role Director's Direct Compensation Director's sex Director's Nationality Director's past employment Director's Qualifications Director's Shared external Directorships Total Company Revenue EBITDA—earnings before interest tax depreciation amortisation Operating Profit Company's net income Company's fixed assets Company's current liabilities Company's long-term liabilities Company's long-term creditors Company's total assets Company's fixed assets Company's current assets Company's total liabilities Company's PE ratio Company's Auditors Company's Financial Advisors Company's Stock Brokers Company's Financial PR Company's Solicitors The central data entry and management location, 100, containing the Data Repository System, 1, acting as the Master Database is connected via a dedicated link to a slave Database system, 200, located at a different location.\",\n \"The Database system, 200, provides the publicly accessible data as a mirror of the data in the Master Database.\",\n \"The Database system, 200, comprises two systems, 300 and 500, each comprising a Database Processor, 4 and 5, connected to a Database, 2 and 3, which act as slaves to the Master Database.\",\n \"There are two database systems to provide redundancy in the case of the failure of one system.\",\n \"The data entered onto the system by means of the Data Repository System, 1, is loaded onto the two database processors, 4 and 5, which perform a series of analytical calculations to transform the raw data into a form that is useful for the system, e.g.\",\n \"the calculation of complex variables, or data set averages and deviations.\",\n \"The two databases, 2 and 3, are then populated with the plurality of predetermined calculated parameters, and the raw data, for storage.\",\n \"In this way the Master Database is kept offline and this mirroring of data takes place regularly, e.g.\",\n \"daily at night when usage is low.\",\n \"This system enables the Master Database to be continuously updated at the central data entry and management location.\",\n \"The two database processors, 4 and 5, are further connected to a data processor 6.The Data Processor 6 is used to perform user end requested calculations, for instance it will calculate peer group parameters, which are discussed hereafter.\",\n \"These parameters are stored in the databases, 2 and 3, and are specific to users.\",\n \"Thus user specific parameters such as Peer Group data are generated and stored in the databases, 2 and 3.The Database Processors, 4 and 5, download the data from the Databases, 2 and 3, and transmit the data to the Data Processor, 6.The Data Processor 6 is connected to two a third system, 500, that comprises two Web Servers, 7 and 8 and a Web Page Data Processor.\",\n \"The system, 500, provides the method of accessing the data in the system, 200, i.e.\",\n \"a web interface.\",\n \"The connection is made across a firewall system for security.\",\n \"Again the existence of two Web Servers is based on the necessity for redundancy in the system.\",\n \"The Data Processor 6, acting as an Application Server to provide data to the Web Servers, 7 and 8, sorts and processes the data into a useable form for the user end of the system, and communicates that data onto the Web Servers, 7 and 8.The Web Servers, 7 and 8, are both connected to a Web Page Data Store, 9.By communicating with the Web Page Data Store, 9, the Web Servers, 7 and 8, generate web pages that are to be communicated to the client's user interface, based on the data received from the Data Processor, 6.The Web Servers, 7 and 8, are connected to an IP network, 10, e.g.\",\n \"the internet.\",\n \"A User's Web Browser, 12, on a User's Computer, 11, is also connected to said IP Network, 10.When requested by the User's Web Browser, 12, the Web Servers, 7 and 8, will transmit said generated web pages to the User's Web Browser, 12, via the IP Network, 10.Also the Web Servers, 7 and 8, can transmit Java Script and Java Applets to control the client's user interface, and to generate a number of the web pages on the User's Web Browser, 12.Thus the user interface can be server side generated, client side generated or a mixture of the two.\",\n \"The User's Web Browser, 12, will then display said web pages in the form of a user interactive display on the User's Computer, 11.The User's Web Browser, 12, can also communicate requests for information back up through the system.\",\n \"This may involve a request for new processed parameters, for example based on a new user selected peer group that the user has set up on his browser.\",\n \"In this scenario a request would be transmitted back to the Data Processor, 6, via the IP network, 10, and one of the Web Servers, 7 or 8.The Data Processor will process the request and transmit the new processed parameters back to the User's Computer in the same manner as the initial parameters were transmitted as discussed above.\",\n \"In this way the Data Processor, 6, generates user specific parameters, e.g.\",\n \"peer groups, and these are stored, along with a User ID, in the databases, 2 and 3, to provide for a level of personalisation when a user accesses the data in the Databases, 2 and 3.User Interface Description In the embodiment of the present invention, the user interface displayed on the user's web browser is split into two main sections, as shown in FIG.\",\n \"2: one supplies information regarding one or more Boards and offers comparative analysis of these Boards against other Boards; and the other offers the same functionality at Director level, allowing the comparison of one or more individual Directors against other Directors.\",\n \"In both cases the system is able to display a large number of parameters for any Board and/or Director through a variety of medium, including tabular form and a number of graphical displays.\",\n \"Data Fields Described by Screen FIG.\",\n \"2 shows the main structure of the system's visual user interface information pages.\",\n \"The pages are organised into a hierarchical structure, with each branch containing further levels of detail below it.\",\n \"As mentioned, above, the user must first select either the Board related section or the Director related section.\",\n \"This structure can clearly be seen in FIG.\",\n \"2 where the Board related pages are on the left hand branch and the Director related pages are on the right hand branch.\",\n \"In each of the two sections the user can select from a list of main topics headings: Summary Associations Remuneration Characteristics Profile Each of these options will display top-level data related to their topic.\",\n \"Some of these topics have further layers of increasingly detailed data, and/or different graphical display methods, in a plurality of related screens and each of the data layers will now be discussed in more detail, initially by looking at the Board related branch and then the Director related branch.\",\n \"Board Summary The Board Summary, as shown in FIG.\",\n \"9, has two themes: firstly it provides a general overview of the members of a Board and displays the following basic information relating to those members: Executive Name- names of all of the Board Directors Role- The position held by the Director in the Company.\",\n \"Time on Board- The length of time the Director has been on the Board (in years).\",\n \"Time in Company- The length of time spent by the Director in the company (in years).\",\n \"Time to Retirement- The length of time to retirement, based on the company's policy retirement age (in years).\",\n \"Stakeholder Representative- specific stakeholder group(s) represented by the Director for example shareholders representatives, union, employees..\",\n \"The second theme to the Board summary is a list of major shareholders in the company, giving their name and an indication as to the size of their holding.\",\n \"Board Associations The Board Associations section has two levels beneath it, as shown in FIG.\",\n \"3, named ‘Single and Multi Associations’, as shown in FIG.\",\n \"10, and ‘Multi Associations’.\",\n \"These two pages provide lists of associations that the Directors of a Board have with the Boards of other companies.\",\n \"A ‘Single Association’ occurs when a Director on the Board of one particular company is also serving on the Board of another company, and is the only person from the subject company to have any association with that latter company.\",\n \"A ‘Multi Association’ occurs when two or more Directors from one particular Board both have associations with another particular Board, i.e.\",\n \"two or more Directors are currently serving on that other Board together, or at least one Director has served on another Board at the same time as at least another Director was also serving on that Board, if at least one of them is currently still sitting on that other board.\",\n \"These two features can be seen on FIG.\",\n \"10, where an example ‘Single Association’ is Georgh Ehrnrooth (bottom row), who is a Director on the Board of Nokia OJY, the subject company, and is also currently a Director on the Board of Wartsila OYJ ABP, and an example of a ‘Multi Association’ is Georg Ehrnrooth and Olli-Pekka Kallasvuo, fifth row from the bottom, who are both on the Board of the Subject company Nokia OYJ and are also both currently on the Board of Sampo OYJ.\",\n \"Board Remuneration The Board Remuneration section can be broken down into five main sections: Remuneration Summary Direct Remuneration—annual cash based pay.\",\n \"Equity Linked Remuneration—annual equity linked pay.\",\n \"Equity Linked Wealth—total equity linked benefits received.\",\n \"Remuneration Ratios This hierarchy can be seen in FIG.\",\n \"4.These sections will now be discussed in turn.\",\n \"The Remuneration Summary, as shown in FIG.\",\n \"4, consists of four separate pages, each displaying the summary information in a different manner.\",\n \"The Remuneration Summary Table, as shown in FIG.\",\n \"11, displays the following parameters: Executive Name- names of all of the Board Directors Role- The position held by the Director in the Company.\",\n \"Total Direct Remuneration- the sum of salary, bonus, pension (defined contribution) and other non-equity remuneration.\",\n \"Total Equity Linked Remuneration- amount of remuneration in the form of equity, shares, and options for a particular financial period.\",\n \"Total Remuneration for the Period- the sum of the Total Direct Remuneration and the Total Equity Linked Remuneration, each as described above.\",\n \"Total Equity Linked Wealth- the value of equity held + the estimated market value of options held.\",\n \"It can also be seen in FIG.\",\n \"11 that the cells in the table are highlighted in a plurality of different colours, this feature gives an indication of the diversity of the data within the table and will be discussed in more detail hereafter.\",\n \"The Remuneration Summary Bubble Chart, as shown in FIG.\",\n \"12, is a chart that plots the three variables from the Remuneration Summary Table, i.e.\",\n \"the Total Direct Remuneration, the Total Equity Linked Remuneration and the Total Equity Linked Wealth onto the same graph providing a simple a quick analysis method for the data.\",\n \"The x-axis on the graph plots the Direct Remuneration value, and the y-axis plots the Total Equity Linked Remuneration values for each individual Director on the Board.\",\n \"Each data point is then expanded in a circular form, wherein the diameter of each circle is proportional to the value of the Total Equity Linked Wealth of each Director.\",\n \"The circles are colour coded to indicate the difference between different types of data, e.g.\",\n \"executive directors, non-executive directors, averages, etc.\",\n \".\",\n \".\",\n \".\",\n \"It can also be seen in FIG.\",\n \"12 that various averages and ‘peer group’ averages are plotted, these will discussed hereafter.\",\n \"The Direct and Equity Remuneration Bar Chart is a standard bar chart, with each Director placed along the x-axis, and the values of their respective Total Direct Remuneration and Total Equity Linked Remuneration for a particular financial period shown as bars, the size of which are relative to a scaled y-axis (for example in £000s of pounds).\",\n \"Lines are also placed on the plot to indicate the values of the Director and peer group averages mentioned above.\",\n \"The Total Equity Wealth Chart is another bar chart showing each Director along the x-axis with a bar extended upwards relative to the y-axis indicating the relative value of the Total Equity Wealth of each Director.\",\n \"Again, some average values are also indicated.\",\n \"The Direct Remuneration page provides a breakdown of the Total Direct Remuneration figures given in the Board Remuneration Summary, for a particular financial period.\",\n \"The display is in the form of a table, with format similar to that of the Remuneration Summary in FIG.\",\n \"11.The parameters displayed are: Executive Name- names of all of the Board Directors Role- The position held by the Director in the Company.\",\n \"Salary- base annual pay Bonus- payments made in addition to salary Pension- employer's contribution to the employee's pension scheme Other- other ad hoc payments, such as relocation and fringe benefits Total Direct- sum of salary, bonus, pension and other income.\",\n \"The Equity Linked Remuneration page provides a breakdown of the Total Equity Remuneration figures given in the Board Remuneration Summary, for a particular financial period.\",\n \"The display is in the form of a table, with format similar to that of the Remuneration Summary in FIG.\",\n \"11.The parameters displayed are: Executive Name- names of all of the Board Directors Role- position each Director holds on the Board.\",\n \"Value of Equity Awarded- value of company shares awarded in the period.\",\n \"LTIP Value- value of Long Term Incentive Plan equity contributions.\",\n \"Intrinsic Value of Options Awarded- the amount by which the current stock price exceeds the exercise price of the options.\",\n \"Note: if the stock price is below the exercise price, there is no intrinsic value.\",\n \"Estimated Value of Options Awarded- Current estimate of the market value of Options gained, based on the latest closing stock price using the Black Scholes option pricing model, or a modification thereof.\",\n \"Total Equity Linked Remuneration the sum of all of the for the Period- other equity linked variables in the table, i.e.\",\n \"the sum of the variables in this list.\",\n \"The Remuneration Ratios page is a series of ratios of some of the remuneration parameters mentioned above.\",\n \"The display is in the form of a table, as shown in FIG.\",\n \"13.The ratios displayed are: Executive Name- names of all of the Board Directors Role- The position held by the Director in the Company.\",\n \"‘Bonus/(Bonus + Salary)’ Ratio ‘Equity Linked/Total Remuneration’ Ratio % Change the percentage change of an individual's Remuneration- remuneration from the one fiscal period to another, e.g.\",\n \"from the ‘99 financial year to the ‘00 financial year.\",\n \"Wealth Delta- the percentage increase in a Directors Wealth due to a 1% increase in a particular company's share price, i.e.\",\n \"a measure of a Director's personal financial gain relative to the company share price success.\",\n \"Board Characteristics The Board Characteristics section can be broken down into five main sections: Roles Network/Biographical Experience Change Diversity as can be seen in FIG.\",\n \"5.These sections will now be discussed in turn.\",\n \"The Board Characteristics ‘Roles’ page provides a breakdown of and parameters relating to the Role that each individual Director has in the company.\",\n \"The page is displayed in a tabular form, similar to that shown in FIG.\",\n \"25 (FIG.\",\n \"25 is the Director Characteristics Roles page, but the format is very similar).\",\n \"The parameters displayed on this Roles page are: Executive Name- names of all of the Board Directors Role- The position held by the Director in the Company.\",\n \"Time to Retirement- The length of time to retirement, based on the company's policy retirement age (in years).\",\n \"Shared External Directorships- The number of Directors on the subject Board who serve on other boards together.\",\n \"Time in Role- number of years the Director has been in his/her current role on Board.\",\n \"Time in Board- number of years the Director has been on the Board, in any role.\",\n \"Time in Company- number of years under employment with company.\",\n \"Time in Sector- number of years the Director has worked in the market sector.\",\n \"The Board Characteristics ‘Network/Biographical’ page provides a breakdown of and parameters relating to the a Director's current Board level commitments, their history of working Board level commitments, and some basic personal details.\",\n \"The page is displayed in a tabular form, as shown in FIG.\",\n \"14.The parameters displayed on this Network/Biographical page are: Executive Name- names of all of the Board Directors.\",\n \"Role- The position held by the Director in the Company.\",\n \"Boards to Date Listed- the total number of Boards of public listed companies on which the Director has served to date including current Boards positions.\",\n \"Boards to Date Unlisted- the total number of Boards of limited companies on which the Director has served to date including current Boards positions.\",\n \"Boards to Date Other- the total number of Other Boards on which the Director has served to date including current Boards positions.\",\n \"Current Boards Listed- the total number of Boards of public listed companies on which the Director currently serves.\",\n \"Current Boards Unlisted- the total number of Boards of limited companies on which the Director currently serves.\",\n \"Current Boards Other- the total number of Other Boards on which the Director currently serves.\",\n \"Average Time in Other the average length of time spent by Companies- individual Directors in companies other than the subject company (in years).\",\n \"Age- the age of the Director Education- the number of qualifications (degree level, inc. professional qualifications) held by the Director.\",\n \"Gender- Shows either the gender of the Director or the proportion of the Directors that are male.\",\n \"Nationality- individual nationalities for each Director, and a nationality mix ratio for the average values (where a value of 1 indicates the most diverse Board possible and a value of 0 indicates a Board comprising all directors of one single nationality).\",\n \"The Board Characteristics ‘Experience’ section consists of two pages, as shown on FIG.\",\n \"5, a Table and a Radar Chart, providing the same information in two different forms.\",\n \"The pages provide a breakdown of and parameters relating to the Experience of the members of the Board, by mixing together some previously discussed parameters from other pages.\",\n \"The pages are displayed firstly in tabular form, similar to that shown in FIG.\",\n \"26, and in graphical form similar to that shown in FIG.\",\n \"27 (FIGS.\",\n \"26 and 27 are the Director Characteristics Experience pages, but the format is very similar).\",\n \"The parameters displayed on the tabular page are: Executive Level- these figures are given for the executives collectively, unlike each individual Director's statistics as before.\",\n \"This column defines between Executive Directors, Non-Executive Directors, and the Board as a whole.\",\n \"Current Boards Listed- the total number of Boards of public listed companies on which the Director currently serves including current Boards positions.\",\n \"Boards to Date Listed- the total number of Boards of public listed companies on which the Director has served to date including current Boards positions.\",\n \"Time in Sector- number of years Director has worked in the market sector.\",\n \"Time on Board- number of years a Director has served on the Board, in any role.\",\n \"Age- the age of the Director Education- the number of qualifications (degree level, inc. professional qualifications) held by the Director.\",\n \"The Board Characteristics Experience Radar Chart is a multi axis chart that will display the parameters displayed in the Experience Table page on one visual chart.\",\n \"The radar charts will be discussed in more detail hereafter.\",\n \"The Board Characteristics ‘Change’ section also consists of two pages, as shown on FIG.\",\n \"5, a Table and a Radar Chart, providing the same information in two different forms.\",\n \"The pages provide a breakdown of and parameters relating to the stability of the Board, by mixing together some previously discussed parameters from other pages and introducing some new parameters from them.\",\n \"The pages are displayed firstly in tabular form, as shown in FIG.\",\n \"15, and in graphical form as shown in FIG.\",\n \"27 (FIG.\",\n \"27 is the Director Characteristics Experience page, but the format is very similar).\",\n \"The parameters displayed on the tabular page are: Executive Level- these figures are given for the executives collectively, unlike displaying each individual Director's statistics as in pages discussed earlier.\",\n \"This column defines between Executive Directors, Non-Executive Directors, and the Board as a whole.\",\n \"Attrition Rate- Proportion of Directors who have left in the previous year.\",\n \"Succession Factor- the standard deviation of the time to retirement.\",\n \"This measures the clustering of Directors and the proximity of Directors to retirement.\",\n \"Standard Deviation of Time on standard deviation from the Board- average value of the time on the Board of all members of each executive group.\",\n \"Average Time in Other standard deviation from the average Companies- value of the time in other companies for all members of each executive group.\",\n \"Direct Remuneration- the average Direct Remuneration for each executive group.\",\n \"Equity Linked Remuneration- the average Equity Linked Remuneration for each executive group.\",\n \"The Change Radar Chart, as for the Radar Chart in the previous section, is a multi axis chart that will display the parameters displayed in the Change Table page on one visual chart.\",\n \"These charts will be discussed in more detail hereafter.\",\n \"The Board Characteristics ‘Diversity’ section also consists of two pages, as shown on FIG.\",\n \"5, a Table and a Radar Chart, providing the same information in two different forms.\",\n \"The pages provide a breakdown of and parameters relating to the diversity of the Board, by displaying the diversity of some previously discussed parameters from other pages.\",\n \"The pages are displayed firstly in tabular form, as shown in FIG.\",\n \"16, and secondly in graphical form as shown in FIG.\",\n \"17.The parameters displayed on the tabular page are: Executive Level- these figures are given for the executives collectively, as with the change statistics above.\",\n \"This column defines between Executive Directors, Non-Executive Directors, and the Board as a whole.\",\n \"Standard standard deviation from the Deviation of average time served on the Board Time on Board- by all of the Directors in each executive group.\",\n \"Standard standard deviation from the Deviation average time served in the of Time in company by all of the Company- Directors in each executive group.\",\n \"Standard standard deviation from the Deviation average time served in the market of Time in sector by all of the Directors in Sector- each executive group.\",\n \"Standard standard deviation from the Deviation of average number of Boards Listed Boards the Directors of each to Date- executive group have served on.\",\n \"Standard standard deviation from the average age of Deviation the Directors of each executive group.\",\n \"of Age- Standard standard deviation from the average Deviation number of qualifications gained by of Education- the Directors of each executive group.\",\n \"Gender- the proportion of males in each executive group.\",\n \"Nationality- a nationality mix ratio where a value of 1 indicates the most nationally diverse Board possible and a value of 0 indicates a Board comprising all directors of one single nationality.\",\n \"The Diversity Radar Chart, as for the charts in the previous two sections, is a multi axis chart that will display the parameters displayed in the Change Table page on one visual chart, as shown in FIG.\",\n \"17.These charts will be discussed in more detail hereafter.\",\n \"Board Profile The Board Profile provides some general contact information for the subject company, e.g.\",\n \"address, phone number, web site etc.\",\n \"It also provides a short list of some basic financial data for the company, such as the Revenue Figures, Profit & Loss Figures and some key ratios, such as the P/E Ratio.\",\n \"Thirdly it provides a list of advisers to the company, such as it's auditors, solicitors and financial advisors.\",\n \"Director Summary The Director Summary, as shown in FIG.\",\n \"22, provides a general overview of the positions held by a Director and displays the following basic information relating to those positions: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.\",\n \"Time on Board- The length of time the Director has been on the Board (in years).\",\n \"Time in Company- The length of time spent by the Director in the company (in years).\",\n \"Time to Retirement- The length of time to retirement, based on the company's policy retirement age (in years).\",\n \"Stakeholder Representative- specific stakeholder group(s) represented by the Director for example shareholders representatives, union, employees..\",\n \"The second theme to the Director summary is a list of major shareholders in the company, giving their name and an indication as to the size of their holding.\",\n \"Director Associations The Director Associations section has two levels beneath it, as shown in FIG.\",\n \"6, named ‘Single and Multi Associations’, as shown in FIG.\",\n \"23, and ‘Multi Associations’.\",\n \"These two pages provide lists of associations that the Director has with other Directors through the Boards he/she serves on.\",\n \"A ‘Single Association’ occurs when a Director on the Director of one particular company is also serving on the Director of another company, and is the only person from the subject company to have any association with that latter company.\",\n \"A ‘Multi Association’ occurs when the Director sits on more than one Board with another Director.\",\n \"Director Remuneration The Director Remuneration section can be broken down into six main sections: Remuneration Summary Direct Remuneration- annual cash based pay.\",\n \"Equity Linked Remuneration- annual equity linked pay.\",\n \"Equity Linked Wealth- total equity linked benefits received.\",\n \"Remuneration Ratios Options, Pensions, and LTIP This hierarchy can be seen in FIG.\",\n \"7.These sections will now be discussed in turn.\",\n \"The Remuneration Summary, as shown in FIG.\",\n \"7, consists of four separate pages, each displaying the summary information in a different manner.\",\n \"The Remuneration Summary Table, which is similar to that shown in FIG.\",\n \"11 (FIG.\",\n \"11 is the Board Remuneration Summary, but the format is very similar), displays the following parameters: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.\",\n \"Total Direct Remuneration- amount of cash based remuneration, for a particular financial period (e.g.\",\n \"a financial year).\",\n \"Total Equity Linked Remuneration- amount of remuneration in the form of equity, shares, and options for a particular financial period.\",\n \"Total Remuneration for the Period- the sum of the Total Direct Remuneration and the Total Equity Linked Remuneration, each as described above.\",\n \"Total Equity Linked Wealth- the value of equity held + the estimated market value of options held.\",\n \"As with the Board Remuneration Summary Table in FIG.\",\n \"11, the Director data tables are highlighted in accordance with the diversity of the data, this will be discussed hereafter.\",\n \"The Remuneration Summary Bubble Chart, similar to that shown in FIG.\",\n \"12 (FIG.\",\n \"12 is the Board Remuneration Summary Bubble Chart, but the format is very similar), is a chart that plots the three variables from the Remuneration Summary Table, i.e.\",\n \"the Total Direct Remuneration, the Total Equity Linked Remuneration and the Total Equity Linked Wealth onto the same graph providing a simple a quick analysis method for the data.\",\n \"The x-axis on the graph plots the Direct Remuneration value, and the y-axis plots the Total Equity Linked Remuneration values for each position that the Director holds.\",\n \"Each data point is then expanded in a circular form, wherein the diameter of each circle is proportional to the value of the Total Equity Linked Wealth of the Director in each position held.\",\n \"The circles are colour coded to indicate the difference between different types of data, e.g.\",\n \"executive Director positions, non-executive Director positions, averages, and ‘peer group’ averages.\",\n \"The Direct and Equity Remuneration Bar Chart is a standard bar chart, with each Director position placed along the x-axis, and the values of their respective Total Direct Remuneration and Total Equity Linked Remuneration for a particular financial period shown as bars, the size of which are relative to a scaled y-axis (for e.g.\",\n \"in £000s of pounds).\",\n \"Lines are also placed on the plot to indicate the values of the Director and peer group averages mentioned above.\",\n \"The Total Equity Wealth Chart is another bar chart showing each Director along the x-axis with a bar extended upwards relative to the y-axis indicating the relative value of the Total Equity Wealth of each Director.\",\n \"Again, some average values are also indicated.\",\n \"The Direct Remuneration page provides a breakdown of the Total Direct Remuneration figures given in the Director Remuneration Summary, for a particular financial period.\",\n \"The display is in the form of a table, with format similar to that of the Board Remuneration Summary in FIG.\",\n \"11.The parameters displayed are: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.\",\n \"Salary- base annual pay Bonus- payments made in addition to salary Pension- employer's contribution to the employee's pension scheme Other- other ad hoc payments, such as relocation and fringe benefits Total Direct- sum of salary, bonus, pension and other income.\",\n \"The Equity Linked Remuneration page provides a breakdown of the Total Equity Remuneration figures given in the Director Remuneration Summary, for a particular financial period.\",\n \"The display is in the form of a table, with format similar to that of the Board Remuneration Summary in FIG.\",\n \"11.The parameters displayed are: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.\",\n \"Value of Equity Awarded- value of company shares awarded in the period.\",\n \"LTIP Value- value of Long Term Incentive Plan equity contributions.\",\n \"Intrinsic Value of Options Awarded- Current Realisable Profit on Options awarded.\",\n \"Estimated Value of Options Awarded- Current estimate of the market value of Options gained, based on the latest closing stock price using the Black Scholes option pricing model.\",\n \"Total Equity Linked Remuneration the sum of all of the for the Period- other equity linked variables in the table, i.e.\",\n \"the sum of the variables in this list.\",\n \"The Remuneration Ratios page is a series of ratios of some of the remuneration parameters mentioned above.\",\n \"The display is in the form of a table, as shown in FIG.\",\n \"24.The ratios displayed are: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.\",\n \"‘Bonus/(Bonus + Salary)’ Ratio ‘Equity Linked/Total Remuneration’ Ratio % Change Remuneration- the percentage change of an individual's remuneration from the one fiscal period to another, e.g.\",\n \"from the ‘99 financial year to the ‘00 financial year.\",\n \"Wealth Delta- the percentage increase in a Directors Wealth due to a 1% increase in a particular company's share price, i.e.\",\n \"a measure of a Director's personal financial gain relative to the company share price success.\",\n \"The Remuneration Options, Pensions and LTIP page provides a revue of these parameters.\",\n \"Director Characteristics The Director Characteristics section can be broken down into three main sections: Roles Network/Biographical Experience as can be seen in FIG.\",\n \"8.These sections will now be discussed in turn.\",\n \"The Director Characteristics ‘Roles’ page provides a breakdown of and parameters relating to each Role held by the Director.\",\n \"The page is displayed in a tabular form, as shown in FIG.\",\n \"25.The parameters displayed on this Roles page are: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.\",\n \"Time to Retirement- The length of time to retirement, based on the company's policy retirement age (in years).\",\n \"Shared External Directorships- The number of Directors who serve on other Boards with the Director.\",\n \"Time in Role- number of years the Director has been on his/her current role on Board.\",\n \"Time on Board- number of years the Director has been on the Board, in any role.\",\n \"Time in Company- number of years under employment with company.\",\n \"Time in Sector- number of years the Director has worked in the market sector.\",\n \"The Director Characteristics ‘Network/Biographical’ page provides a breakdown of and parameters relating to the a Director's current Board level commitments, their history of working Board level commitments, and some basic personal details.\",\n \"The page is displayed in a tabular form, similar to the Board Characteristics Network/Biographical page as shown in FIG.\",\n \"14.The parameters displayed on this Director Network/Biographical page are: Executive Name- names of the companies with whom the Director has a Board level position Role- The position held by the Director on each Board.\",\n \"Boards to Date Listed- the total number of Boards of public listed companies on which the Director has served to date including current Board positions.\",\n \"Boards to Date Unlisted- the total number of Boards of limited companies on which the Director has served to date including current Board positions.\",\n \"Directors to Date Other- the total number of Other Boards on which the Director has served to date including current Board positions.\",\n \"Current Boards Listed- the total number of Boards of public listed companies on which the Director currently serves.\",\n \"Current Boards Unlisted- the total number of Boards of limited companies on which the Director currently serves.\",\n \"Current Boards Other- the total number of Other Boards on which the Director currently serves.\",\n \"Average Time in Other the average length of time spent by Companies- individual Directors in companies other than the subject company (in years).\",\n \"Age- the age of the Director Education- the number of qualifications (degree level, inc. professional qualifications) held by the Director.\",\n \"Gender- Shows the gender of the Director.\",\n \"Nationality- shows the nationality of the Director.\",\n \"The Director Characteristics ‘Experience’ section consists of two pages, as shown on FIG.\",\n \"8, a Table and a Radar Chart, providing the same information in two different forms.\",\n \"The pages provide a breakdown of and parameters relating to the Experience of the Director, by mixing together some previously discussed parameters from other pages.\",\n \"The pages are displayed firstly in tabular form, similar to that shown in FIG.\",\n \"26, and in graphical form as shown in FIG.\",\n \"27.The parameters displayed on the tabular page are: Executive Name- names of the companies with whom the Director has a Board level position Current Boards Listed- the total number of Boards of public listed companies on which the Director currently serves including current Board positions.\",\n \"Boards to Date Listed- the total number of Boards of public listed companies on which the Director has served to date including current Board positions.\",\n \"Time in Sector- number of years Director has worked in the market sector.\",\n \"Time on Board- number of years a Director has served on the Board, in any role.\",\n \"Age- the age of the Director Education- the number of qualifications (degree level, inc. professional qualifications) held by the Director.\",\n \"The Director Characteristics Experience Radar Chart is a multi axis chart that will display the parameters displayed in the Experience Table page on one visual chart.\",\n \"The radar charts will be discussed in more detail hereafter.\",\n \"Director Profile The Director Profile provides some general personal information for the Director, e.g.\",\n \"date of birth, nationality etc.\",\n \"It also provides a short CV showing the positions that he/she currently holds and positions that the Director has held in the past, with a brief description of each of those positions.\",\n \"User Interface Having looked at the structure of the information pages displayed on the user's web browser, the method by which the user navigates around these pages and how they select which data is to be displayed will now be described.\",\n \"FIG.\",\n \"9 shows an example of the general layout of the user interface.\",\n \"The display comprises two parts: a fixed thin navigation pane on the left hand side of the display which displays the list of available information page windows that a user can select, these were discussed above, as well as other features such as a search facility and some historical selection lists, that will be discussed hereafter; the second part of the display comprises a changeable information display pane which can display a series of web style pages that each provide a different set of the actual Board/Director related information pages that are received from the web server.\",\n \"A user can select which page of data he wishes to have displayed in the right hand pane by selecting from the hierarchical list in the left hand navigation pane.\",\n \"The navigation pane provides a lower level break down of available pages as the user navigates around the system.\",\n \"For instance in FIG.\",\n \"9 the top level structure of the system, as shown in FIG.\",\n \"2, is visible, whereas in FIG.\",\n \"11, where the user has selected the ‘Board Remuneration Summary’ screen, it can be seen that the navigation list has been extended to show the levels below that of the screen which is being displayed, hence allowing the user to access different methods of display for the ‘Board Remuneration Summary’ information; ‘Table’, ‘Bubble Chart’, ‘Direct and Equity Chart’ and ‘Equity Wealth Chart’ in this case.\",\n \"Once the user has selected either the Board section or the Director section, by clicking on one of the two labelled tabs in the top left hand corner of the screen, they have to next choose a particular Board/Director to be used as the subject of the data.\",\n \"This selection will instruct the system to download all relevant information for that Board/Director to then allow the user to navigate around the different screens in the navigation panel viewing different data sets related to that Board/Director.\",\n \"This Board/Director selection is conducted by using one of two search systems which are situated in the navigation pane.\",\n \"FIG.\",\n \"9 shows these search facilities situated below the screen navigation list in the navigation pane on the left hand side of the display.\",\n \"One search option, the first box below the screen navigation list, comprises a manual entry search system which allows the user to enter some data which the system then uses to search the database.\",\n \"Another search option is called the ‘Advanced Search’ and is situated in the box below the manual search data entry boxes, this involves the user defining a search by selecting from a number of parameters in a pre-defined list that the system will use to narrow the search of the database.\",\n \"The manual search facility offers two methods of data entry for the search; a user can either input a company/person's name into the ‘Company Name’ text box displayed on the navigation pane; or the user can input another form of company related designation data, such as the SEDOL CODE—a financial markets company ID (the data type can be selected from a drop-down box), by inputting data into the ‘Company ID’ text box on the navigation pane.\",\n \"The system will then conduct a search of the database and produce a search results list such as those shown in FIGS.\",\n \"21 and 31.The user can then select any Board/Director he wishes to use as the subject of the analysis in the information display pages by clicking on a Board/Director's name from the search results list.\",\n \"The system will then retrieve the parameters for that chosen Board/Director and initially display the ‘Summary’ page for that subject Board/Director, as shown in FIG.\",\n \"3.The Advanced Search facility is initiated when the user clicks on the ‘Advanced Search’ button in the navigation pane.\",\n \"A screen such as that in FIG.\",\n \"20 will be displayed for an advanced Board search, and such as that in FIG.\",\n \"30 for and advanced Director search.\",\n \"The user will then select at least one of the pre-defined parameters, such as the country or sector of the Board/Director they wish to search for and/or by inputting at least one piece of data into any one of the data entry fields, such as defining a Market Capitalisation of the company of the Board they wish to search for.\",\n \"The system will then conduct a search of the database based on the data entered by the user and produce a search results list, again such as those shown in FIGS.\",\n \"21 and 31.The user can then select any Board/Director he wishes to use as the subject of the analysis in the information display pages by clicking on a Board/Director's name from the search results list, the system will then retrieve the parameters for that Board/Director and initially display the ‘Summary’ page for that subject Board/Director, as shown in FIG.\",\n \"9.A third method of choosing a subject Board/Director to be the subject of the information pages is to select a name from the ‘Viewed Boards’ list at the bottom of the navigation pane.\",\n \"The system will store a list of the last n subjects to have been chosen by the user and display them at the bottom of the navigation pane.\",\n \"This allows the user to easily return to a recent set of displayed data without having to pass through either of the two search methods again.\",\n \"Once a subject Board/Director has been chosen and its data downloaded and displayed, the name of that Board/Director will be displayed in the top left-hand corner of the display on all of the information pages.\",\n \"When a new Board/Director is chosen, the name displayed will be updated accordingly.\",\n \"Many of the information pages display the information in tabular form, and there is a marked difference between the style of these tables for the Board section and the Director section in terms of how the data is displayed, these two different styles will now be explained, firstly with reference to Boards.\",\n \"The rows of the Board related tables of data are split up into three main categories, the top category comprising the Executive Directors, the second category comprising the Non-Executive Directors and the third category comprising All Positions.\",\n \"FIG.\",\n \"11 shows how this structure is displayed.\",\n \"The top category of FIG.\",\n \"11 lists the names of the Executive Directors of ‘Barclays’, the second category lists the names of all of the Non-Executive Directors of Barclays, and the bottom category contains the average values for all of the Directors together.\",\n \"It should be noted that this format allows the user to analyse the data for every individual on the selected subject Board, as well as the average value for the ‘executives’ category, an average value for the ‘non-executives’ category and the average value for all members of the Board collectively.\",\n \"Thus, this set-up provides a large amount of segregated and averaged data to allow the user to analyse a detailed view of all aspects of a Board at the same tine.\",\n \"The Director section display is shown in FIG.\",\n \"25.It can be seen that the selected Director is Tessen von Heydebreck and that he serves on two Boards, as an Executive Director with Deutsche Bank, and a Non-Executive Director with BASF AG.\",\n \"In a similar manner to the Board tables, these different positions are split in to separate categories.\",\n \"Any Executive positions will be displayed in the top category, and any Non-Executive positions in the second category, with an average for all positions held shown in the bottom category.\",\n \"Hence, all aspects of an individual Director's commitments are displayed at one time, as well as an average for those positions, providing a comprehensive analysis platform without having to look up each position individually.\",\n \"Peer Groups One of the features of the embodiment of the present invention are the so called ‘Peer Groups’ that allow the user to compare a selected subject Board/Director with one or more corresponding Boards/Directors.\",\n \"These Peer Groups act as a benchmark with which to compare and contrast different Boards/Directors, they can comprise one single Board/Direct to allow a direct comparison between two particular companies/individual Directors, or they can contain a plurality of different Boards/Directors, grouped for example by sector.\",\n \"As mentioned above, the tables of data for both the Board and Director sections are separated into three categories, and as well as showing data for the individual Directors of a Board or the individual Positions held by a Director, each of these categories also displays an average value for all data within that category.\",\n \"Below these average values, as can be seen in FIGS.\",\n \"11 and 25, is displayed an average value for all of the members of a Peer Group, this can be used as a comparative measure for the individual Board members/Director positions, as well as for the average values for each category.\",\n \"A key function of these Peer Groups is to enable the user to define the constituents of any Peer Group and can select any Peer Group from their records to be used in the comparison.\",\n \"At the bottom of the navigation pane, above the Viewed Boards/Directors section is the Peer Group Button which when selected opens up the Peer Group management screens.\",\n \"These screens work in exactly the same way for the Board sections as for the Director section and will therefore be explained together, with reference to a number of corresponding diagrams.\",\n \"FIGS.\",\n \"18 and 28 show the first page of the Peer Group management pages.\",\n \"This page displays all of the Peer Groups that have been set up by the user to date.\",\n \"We can see in FIG.\",\n \"18 that the user has set up three Peer Groups: PG1, PG2 and PG3, and that those Peer Groups contain seven, three and two members respectively.\",\n \"From the Peer Group amendment screen the user is able to view the members of any Peer Group, amend the members of any Peer Group, or set up a new Peer Group.\",\n \"To view the members of a Peer Group, shown in FIGS.\",\n \"18 and 28, the user simply clicks on the name of one of the Peer Groups and a screen containing a list of Peer Group members is displayed, such as those shown in FIG.\",\n \"19 for the Board Peer Group PG1 and in FIG.\",\n \"29 for the Director Peer Group PG2.Hence it can be seen that the Board Peer Group PG1 consists of seven Boards, including those of Barclays and Marks & Spencer, and the average parameter values displayed on a table of data on a Board information screen, for this Peer Group, would be equal to the average value for all of the groups in the Peer Group.\",\n \"The user can easily amend the members of a Peer Group whilst in the Peer Group members screen, as in FIG.\",\n \"19 or 29, by adding a new member or deleting an existing Peer Group member, as well as being able to move or copy a member to another Peer Group.\",\n \"To delete a member the user would check the check box to the left of any current Peer Group member and then click on the delete button in the top left hand corner of the screen.\",\n \"To copy or move a member the same process would be used except that having selected a member to copy or move the user would choose another Peer Group from a drop-down menu in the top right corner, once a member and other Peer Group have been chosen the user clicks either the copy or move button and the action is completed.\",\n \"Adding a new user to a Peer Group is slightly more complicated and requires the use of the two searching tools, manual and advanced, that were discussed earlier.\",\n \"A user can search as described earlier by company name, company ID or by using the advanced search and a search results page is displayed, as in FIGS.\",\n \"21 and 31.If the Board/Director is present in the search result list, the user will check the check box next to the displayed name and then select a Peer Group from a drop-down menu in the top right hand corner of the screen into which they wish to place Board/Director.\",\n \"Once a subject and Peer Group have been chosen, the user clicks on the ‘Add To Peer Group’ button above the search results list and the action is duly performed.\",\n \"Setting up a new Peer Group mainly involves a combination of the features that have already been discussed, once the initial naming procedure has been carried out.\",\n \"In the main Peer Group page, FIGS.\",\n \"18 and 28, the user can set up a shell for a new Peer Group by typing a name into the text box next to the ‘New’ button above the list of current Peer Groups.\",\n \"Once a name has been entered and the ‘New’ button has been selected, the shell is created and the name of the new Peer Group are displayed along with the other current Peer Groups.\",\n \"The user can then enter that new Peer Group's member screen and add new members, or indeed copy/move members from other group's member screen to the new Peer Group.\",\n \"Once one or more Peer Groups have been set up, the user is able to use the Peer Group comparison facility in one of the main Board/Director information screens.\",\n \"Most of the tabular and graphical screens contain a combination of a series of drop-down menus in the top right hand corner of the screen.\",\n \"These enable the user to perform one of three actions: choose a Peer Group to act as a comparison; choose and individual member of any one Peer Group to act as a comparison, or add the current subject Board/Member to a Peer Group.\",\n \"When the user selects a Peer Group from the ‘Compare to Peer Group’ drop-down list, the system will display the average value for all members of that Peer Group for all of the parameters in any information table or graph.\",\n \"For instance, in FIG.\",\n \"14, the Compare to Peer Group display box in the top right hand corner of the screen is indicating that Peer Group PG1 has been selected for the comparison to Nokia OYJ, the subject Board, named in the top left hand corner of the screen.\",\n \"The Peer Group averages in the table refer to the average value of all of the members of Peer Group PG1.If the user wished to compare Nokia against a different Peer Group they are able to select a different Peer Group name from the Compare to Peer Group selection drop down menu.\",\n \"The user is able to compare the subject Board, Nokia OYJ in the example in FIG.\",\n \"14, against a particular member of the selected Peer Group, PG1 in this example, by selecting one of the members of PG1 from the ‘Compare to Peer Member’ drop-down list, below the ‘Compare to Peer Group’ list in the display.\",\n \"The user could, for instance, choose ‘Marks & Spencer's’ from the PG1 Peer Group list, this would then display the average values for the Marks & Spencer's Board on the table of data and hence allow the user to compare the Boards of Nokia OYJ with Marks & Spencer's directly.\",\n \"The third drop-down menu in the top right hand corner of the screen allows the user to add the current subject Board, Nokia OYJ in the example of FIG.\",\n \"14, to any current Peer Group.\",\n \"The user simply clicks on the drop-down menu and a list of existing Peer Groups is displayed, the user can then select one of these per groups and the subject Board is added to that Peer Group automatically.\",\n \"The Peer Group system works in exactly the same manner for the Director information screen as explained above for the Board information screens.\",\n \"The only difference being that the Peer Groups are made up of individual Directors and the ‘Compare to Peer Member’ list will offer a list of said individuals to allow the user to compare the subject Director to any one specific other Director.\",\n \"It should be noted that the Peer Group comparisons are also use in the graphical information pages.\",\n \"For instance their values are plotted on the bubble charts for comparison purposes, as mentioned earlier.\",\n \"Also, they are used as a comparative means on the radar charts, FIG.\",\n \"17 shows an example of this where the thin line is that of the selected Board, Royal Bank of Scotland, and the shaded area represents the values for the selected Peer Group PG1.Again these pages have the functionality to change the Peer Group, view an individual member of a Peer Group or add the subject Board to a Peer Group.\",\n \"Colour Coding Another feature of the embodiment of the current invention, as mentioned earlier, is the highlighting of the cells of the tables of information to visually indicate the diversity of the data.\",\n \"Again the Peer Groups play an important part in this function as the reference point from which the diversity is calculated.\",\n \"FIG.\",\n \"14 shows a table of data for the Network/Biographical parameters.\",\n \"Displayed above the table is a ‘Variance key’ that indicates the association between the colour of the highlighting of each cell and the measure of variance of the data in the cell, as compared to its corresponding Peer Group average value.\",\n \"The cells are highlight in relation to the number of standard deviations that the data contained therein is away from the average value of the selected Peer Group.\",\n \"If the data is within ±1 standard deviations of the Peer Group average, the cell will not be highlighted.\",\n \"If the cell is between −1 and −2 or +1 and +2 standard deviations of the average of the Peer Group it is highlighted in a light colour, and so on where the highlighting becomes darker with the more standard deviations the data is away from the Peer Group average.\",\n \"This feature provides easily recognisable information to a user.\",\n \"Patterns and trends can be identified in the data much more easily using this feature.\",\n \"It removes the need for a user to laboriously read every data parameter in the tables in order to analyse the data.\",\n \"Radar Charts Another feature of the embodiment of the current invention is the radar charts, and especially the method used to determine the scale of the axes.\",\n \"FIG.\",\n \"17 shows a Diversity Radar Chart for the Royal Bank of Scotland and Peer Group PG1.This radar chart is the corresponding graphical representation of the data in the Diversity Table in FIG.\",\n \"16.The first aspect to note is the number of columns of data within the Diversity Table and the corresponding number of axes on the Diversity Chart.\",\n \"The chart will display as many parameters from the table as it has data for.\",\n \"In many cases the Diversity table is fully populated with data and the Diversity Chart will have a potential 8 axes.\",\n \"However, if there is insufficient data available, i.e.\",\n \"there is no data for the subject Board or there is no data for the selected Peer Group, the radar chart is adjusted and the axis related to the parameter with insufficient data is removed and hence not displayed.\",\n \"It can be seen from FIGS.\",\n \"16 and 17 that there is only sufficient data in four of the parameter columns of the Diversity Table, the Gender column does contain some data but is not fully populated, hence the corresponding Radar Chart only has four axes.\",\n \"The scaling of the axes is based on a similar method to that of the highlighting mentioned above, in that they are bound by a range of standard deviations.\",\n \"The axes are bound by a range of ±3 standard deviations from the average of a Peer Group.\",\n \"Hence the minimum value of each axis on the Radar Chart equals −3 standard deviations from the average of a Peer Group and the maximum value of each axis equals +3 standard deviations from the average of a Peer Group.\",\n \"In this particular embodiment of the invention the Peer Group used for this scaling is not the user selected Peer Groups, it comprises all of the Boards/Directors that are based in the same country as the subject Board/Director.\",\n \"For example in FIG.\",\n \"17, the Royal Bank of Scotland is based in the UK, hence the scale of the axes in the corresponding Diversity Chart are ±3 deviations from the average value for all other Boards on the Database that are also based in the UK.\",\n \"This feature provides a useful method of displaying the data to allow easy analysis by a user.\",\n \"The multi axis plot facilitates comparative analysis of a multitude of parameters related to Boards and/or Board members.\",\n \"Date Change Another feature of the embodiment of the invention is the date change facility.\",\n \"On each information display page there is a date in the top left corner, below the subject Board/Director name, as can be seen in FIGS.\",\n \"16 and 17 for example.\",\n \"The date displayed in the box denotes the point in time to which the information displayed relates.\",\n \"The user is able to change this date, by selecting another date from the drop down menu, hence allowing them a very simple tool to view historical data across a wide date range.\",\n \"Mouse Over Another feature of the embodiment of the invention is the ‘mouse-over’ facility.\",\n \"The system displays small information notes next to a mouse pointer as the user moves around the screen to enable the user to quickly understand the functionality of a particular feature on the display.\",\n \"Additional to this is a mouse-over feature to aid the colour coding of data tables facility discussed above.\",\n \"As a user passes their mouse over a shaded cell in a table of data a small information note is displayed showing the specific value of deviation that the particular parameter has from the Peer Group average value, this feature adds more detail to the colour coding system discussed above, when it is required by the user.\",\n \"Pre-Defined Peer Groups In another embodiment of the current invention a plurality of pre-defined Peer Groups are be loaded onto the database.\",\n \"The user is able to use, select, and even amend, these pre-defined Peer Groups in the same way as the current user defined Peer Groups, discussed above.\",\n \"These pre-defined Peer Groups are organised into logical groups, such as by sector or geographic region and allows the user to compare a subject Board/Director against one of these default Peer Groups without having to set-up a Peer Group themselves.\",\n \"Fantasy Board In another embodiment of the current invention the Board and Director sections are mixed together and function as one unit displaying data for both individual Directors as well as Boards in the same information tables and on the same information charts.\",\n \"This allows for analysis of Boards against Directors, e.g.\",\n \"a user can set up a ‘fantasy Board’ by setting up a Peer Group of Directors and then compare that fantasy Board against a real Board.\",\n \"Or, in turn, an individual Director can be compared against the average value for a Peer Group of one or more Boards.\",\n \"Enhanced Board/Member Associations Functionality In another embodiment of the current invention the Board/Director Associations information pages will not only display links between the members of the subject Board/subject Director and other positions that they may hold, but also associations from a subject Board/Director to a target Board Director, via a plurality of other Boards/Directors Associations.\",\n \"For Example, this will allow the user to input their subject Board/Director and a ‘target’ Board/Director with which they would like to make contact.\",\n \"The system will search the database and display a chain of associations which link the two user selected Boards/Directors together.\",\n \"This display may provide a search result such as: Board 1 is linked to Board 3 via Mr. A; Board 3 is linked to Board 5 via Mr. B; Board 5 is linked to Board 2 via Ms. C; Where Board 1 is the ‘subject’ Board and Board 2 is the ‘target’ Board.\",\n \"This functionality allows the user to build networks of Boards and Directors for use in a plurality of business situations.\",\n \"Company Advisors Associations In another embodiment of the current invention, the associations functionality is extended to encompass the advisors of companies, a list of which is already provided on the Board Profile information page for each Board.\",\n \"In this embodiment a user is able to conduct a search of Associations based on a particular advisor, i.e.\",\n \"make the advisor the subject of the information and display the associations for that advisor.\",\n \"For example, if a particular company has been in trouble as a result of one of their advisors, a user can conduct a search of the Associations of that advisor to other Boards and the system would display a list of any other Boards that use the same advisor.\",\n \"This is useful, for example in the field of fund management, where an advisor has recently shown weakness whilst acting for one of a Fund Manager's holdings, that fund manager could perform a due diligence check to ascertain if any of his other shareholdings are advised by the same advisor by conducting an Associations search on that advisor.\",\n \"Board Summary ‘Timeline’ In another embodiment of the current invention a ‘Board Timeline’ chart, as that shown in FIG.\",\n \"32, is displayed on a second information display page under the ‘Board Summary’ section.\",\n \"This information page displays a bar chart style chart over a discrete time period, pre-set or user defined, showing bars to represent the periods during which each Director has been on the Board, and the position they held, during the said time period.\",\n \"Director Associtaion ‘Multi’ In another embodiment of the current invention a graphical bar chart style plot is displayed to provide a more user friendly method of viewing the Director Multi Associations.\",\n \"An example of this graphical information page is in FIG.\",\n \"37.The names of each Director are placed along the y-axis of a bar chart, with time (in years) along the x-axis.\",\n \"A bar is then plotted alongside each Director's name for a period in which they held a Director level role, colour coding is used to differentiate between the different companies on which they served.\",\n \"This chart provides a much more user friendly method of reviewing the multi associations of a number of Directors.\",\n \"Analyses Topic Section In another embodiment of the current invention the Board and Director sections comprise a sixth main topic heading, in addition to the five existing main sections as discussed above and shown in FIG.\",\n \"2, named ‘Analyses’.\",\n \"This extra section is added to the user interface navigation pane, as shown in the navigation pane on the left hand side of FIGS.\",\n \"32 and 37, between the Characteristics and Profiles sections.\",\n \"The two sections contain different sub-layers, and will now be discussed in turn, beginning with the Board section.\",\n \"Board Analyses The Board Analyses section can be broken down into five main sections: Board Share Price History Board and Shareholder Returns Board Time Analysis Board Experience over Time Options and Equity Scenarios Each of these sections will now be discussed in turn.\",\n \"Board Analyses ‘Board Share Price History’, as shown in FIG.\",\n \"33, displays the share price of the subject company over a discrete time period, pre-set or user-defined, with major Board related corporate actions indicated graphically on top of the chart.\",\n \"For example, if a CEO resigns, a mark will be placed on the graph to indicate the point in time when this occurs.\",\n \"This data can be loaded onto the database automatically or manually, as discussed above, via the analytics data entry method on the Data Repository System (1, FIG.\",\n \"1) at the Central Data entry Management Location (100, FIG.\",\n \"1).\",\n \"This facility enables the user to analyse the fluctuations in the company share price relative to such Board related corporate events and allows them to gain a fuller understanding of share price volatility at such points in time.\",\n \"Hence this embodiment offers the user a tool to assess the importance of Board stability for a particular company and hence gain an insight into future share price fluctuations on the announcement of a Board related corporate action.\",\n \"Board Analyses ‘Board and Share Holder Returns’, as shown in FIG.\",\n \"34, displays a mix of different data types on one graphical display, including Director Direct Remuneration, Director Equity remuneration, Company Income and the Company Share Price.\",\n \"The main element of the graph is a bar chart with two bars for each year along the x-axis, one of these bars displays information relating to the remuneration (further broken down into sections of: salary, other direct, equity, options and LTIP) of the Directors of the Board during that year, the other showing the Net Income of the Company in the same year.\",\n \"On top of the bar chart is displayed the share price of the subject company over the discrete time period, pre-set or user-defined, displayed.\",\n \"This graph enables the user to analyse a number of different features at once, for instance the relationship between company net income and the share price, or that of the Directors income in relation to the share price, plus many more.\",\n \"Board Analyses ‘Board Time Analysis’, as shown in FIG.\",\n \"35, graphically displays a summary of the external commitments of the members of a Board.\",\n \"The graph comprises a bar chart with three bars displayed in each year along the x-axis, each bar to represent board-level positions held on ‘listed’, ‘unlisted’ and ‘other’ boards respectively.\",\n \"The y-axis provides a measure of the average number of those board-level positions held by the members of the subject board during each particular year.\",\n \"This graph is a graphical representation of some of the information given in the Board Characteristics ‘Network/Biographical’ section discussed earlier.\",\n \"Board Analyses ‘Board Experience over Time’, as shown in FIG.\",\n \"36, displays a bar chart offering a summary of the experience of the members of the Board on a yearly time basis.\",\n \"Each year plotted along the x-axis contains two bars: the first of which provides an average value, for all of the Board members, of the time spent in the current company, and a measure of how much of that time was spent serving on the Board; the second bar represents the average number, for all the members of the Board, of all the Boards on which they currently serve and all the Boards on which they have served to date.\",\n \"This is another graphical representation related to the experience parameters displayed in the Board Characteristics ‘Experience’ Table information page.\",\n \"Board Analyses ‘Options and Equity Scenarios’ allows the user to assess the change in the value of the Board held options and equity by running scenarios related to the stock price of a company.\",\n \"The user can enter parameters for an imaginary rise or fall in the price of a stock and the system will calculate and display the effect that this change in stock price would have on the option and equity valuation of the Board of that company.\",\n \"Director Analyses The Director Analyses section can be broken down into two main sections: Director Share Price History Options and Equity Scenarios These sections will now be discussed in turn.\",\n \"Director Share Price History, as shown in FIG.\",\n \"38, is a tool to allow the user to assess the impact of a Director on the company share price during each of his Director level positions.\",\n \"A discreet time period, pre-set or user selected, is used and a separate share price chart for each of the companies on whose Board the Director has served during that time are arranged on the same screen.\",\n \"On top of these share price graphs will be displayed time related information relating to when the Director served with each company and the corresponding role which he held on each Board.\",\n \"This data, as with the Board time line analysis data is entered onto the system using the manual analytics software package on the Data Repository System (1, FIG.\",\n \"1) at the Central Data entry Management Location (100, FIG.\",\n \"1).\",\n \"By reviewing the information displayed the user is able to quickly gain an impression of the historical impact of the arrival and departure of the Director in question at each of his past companies, as well as the performance during his/her time on the Board.\",\n \"Hence, the system is able to provide an indicative historical performance analysis for the Director.\",\n \"Director Analyses ‘Options and Equity Scenarios’ allows the user to assess the change in the value of the Director held options and equity by running scenarios related to the stock price of a company.\",\n \"The user can enter parameters for an imaginary rise or fall in the price of a stock and the system will calculate and display the effect that this change in stock price would have on the option and equity valuation of a Director of that company.\",\n \"Peer Group Alerts In another embodiment of the current invention The Peer Group Alerts section forms part of the Board Peer Groups Section discussed above.\",\n \"These alerts target allow the user to receive targeted corporate action based information about a Board and/or Director Peer Group as follows: Board Peer Group Alerts allows the user to define a list of criteria for all the members of a Peer Group of Boards, on the action of which an alert will be flagged up to the user indicating the event that takes place.\",\n \"FIG.\",\n \"39 shows an example of the definitions screen, it can be seen that there is a wide range of Director level and Board level criteria which can be defined for the members of the Peer Group that has been selected from the drop-down at the top of the screen.\",\n \"If any one of the pre-defined variables is met, for instance a Chairman of one of the Boards within the Peer Group leaves, or the average age of the Boards in the Peer Group rises above the selected level, then a message indicating the event will be supplied to the user, either by e-mail or on login depending on the user's preference, as shown in FIG.\",\n \"40.This facility allows the user to target the corporate news that they require.\",\n \"Instead of sifting through large quantities of literature to find the information that is useful to them, the user can define the areas in which they are interested, by using the screen shown in FIG.\",\n \"39, for the Boards in which they are interested, by setting up a Peer Group containing those Boards, as discussed above, and hence receive news that is personally targeted at the user.\",\n \"Director Peer Group Alerts allows the user to define a list of criteria for all the members of a Peer Group of Directors, on the action of which an alert is flagged up to the user indicating the event that takes place.\",\n \"FIG.\",\n \"41 shows an example of the definitions screen, it can be seen that there is a wide range of Director level criteria which can be defined for the members of the Peer Group that has been selected from the drop-down at the top of the screen.\",\n \"If any one of the pre-defined variables is met, for instance a Director within the Peer Group leaves his position, or the average Direct Compensation within the Peer Group rises above the selected level, then a message indicating the event is supplied to the user, either by e-mail or on login depending on the user's preference, as shown in FIG.\",\n \"42.This facility allows the user to target the corporate news that they require.\",\n \"Instead of sifting through large quantities of literature to find the information that is useful to them, the user can define the areas in which they are interested, by using the screen shown in FIG.\",\n \"41, for the Directors in which they are interested, by setting up a Peer Group containing those Directors, as discussed above, and hence receive news that is personally targeted at the user.\",\n \"Although the present invention has been described hereinbefore with reference to specific embodiments, it will be apparent to a skilled person that modifications lie within the spirit and scope of the present invention.\",\n \"For example, although the system has been described as being executed on a network, it could be implemented on a single computer or any suitable computer system.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476975"}}},{"rowIdx":990,"cells":{"text":{"kind":"list like","value":[["Polypeptide serving as angiogenic marker and dna thereof","A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease and cancer, the method of detection of the marker and a diagnosis kit of the diseases are provided.","Additionally, therapeutic agents of the diseases are provided.","The expression of KIAA1036 is enhanced in ovarian cancer and large bowel cancer and KIAA1036 expresses in umbilical vein endothelial cell and inhibits DNA synthesis in the cells, cell migrating and lumen formation.","Therefore KIAA1036 is useful as a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.","Additionally, KIAA1036 is useful for screening of agonists, antagonists, DNA synthesis inhibitors, cell migrating inhibitors and neovascular inhibitors.","The substances obtained by the screening, KIAA1036 and the antibodies are useful as therapeutic agents the above disease."],["1.A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof.","2.A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polynucleotide hybridizing under a stringent condition with a polynucleotide having a base sequence as set forth in C of 1-C of 1480 of SEQ ID NO: 1 or fragment thereof.","3.A marker of claim 1, which contains a polynucleotide having a base sequence as set forth in C of 1-C of 1480 of SEQ ID NO: 1 or fragment thereof.","4.A marker of claim 1, which contains a polynucleotide having a base sequence as set forth in C of 1-C of 5481 of SEQ ID NO: 1 or fragment thereof.","5.A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.","6.A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polypeptide wherein 1 or more amino acid residue(s) has a mutation(s) selected from deletion, substitution and addition in an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.","7.A marker of claim 5, which contains a polypeptide consisting of an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.","8.A method of detection of a marker of any one of claims 1-7 from a sample derived from human.","9.A method of detection or assay of a marker of any one of claims 1-4, which comprises the following processes and using a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof, (a) a process that the polynucleotide and a sample are contacted, and (b) a process that a polynucleotide binding with the polynucleotide is detected.","10.A method of detection or assay of a marker of any one of claims 1-4, which comprises the following processes and using a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof, (a) a process that cDNA is prepared from a sample, and (b) a process that cDNA is amplified with the polynucleotide fragment and detected.","11.A method of detection or assay of a marker of any one of claims 5-7, which comprises the following processes and using an antibody recognizing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (a) a process that the antibody and a sample are contacted, and (b) a process that the binding between the antibody and a polypeptide is detected.","12.A method of detection of a disease relating to a marker of any one of claims 1-7, which comprises using a method of any one of claims 9-11.13.A method of detection of claim 12, wherein the disease is vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.","14.A diagnostic kit of a disease relating to a marker of any one of claims 1-7, which comprises using a method of any one of claims 9-11.15.A diagnostic kit of claim 14, wherein the disease is vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.","16.A screening method of a binding substance of a marker of any one of claims 5-7, which comprises the following processes, (a) a process that a marker of any one of claims 5-7 and a subject are contacted, and (b) a process that the binding between the marker and the subject is detected.","17.A kit for screening of a binding substance of claim 16, which contains a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof.","18.A screening method of a binding activity regulatory substance which regulates a binding between a marker of any one of claims 5-7 and a binding substance of the marker, which comprises the following processes, (a) a process that a marker of any one of claims 5-7 and a binding substance of the marker are contacted in the presence or absence of a subject, and (b) a process that a binding activity of the marker and the binding substance is compared in the presence or absence of a subject.","19.A kit for screening of a binding activity regulatory substance of claim 18, which contains a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof.","20.A screening method of an expression regulatory substance of a marker of any one of claims 1-7, which comprises the following processes, (a) a process that a cell, which can express a marker of any one of claims 1-7, and a subject are contacted, and (b) a process that a change of an expression level of the marker is detected by using a method of any one of claims 9-11.21.A kit for screening of an expression regulatory substance of a marker of any one of claims 1-7, which comprises using the method of claim 20.22.A DNA synthesis inhibitor containing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.","23.A cell migrating activity inhibitor containing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.","24.A pharmaceutical composition which contains at least one selected from a DNA synthesis inhibitor of claim 22, a cell migrating activity inhibitor of claim 23, a polynucleotide having a base sequence as set forth in SEQ ID NO: 1 or fragment thereof, a human gene therapeutic expression vector containing polynucleotide having a base sequence as set forth in SEQ ID NO: 1 or fragment thereof, a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof and an antibody to the polypeptide.","25.A pharmaceutical composition of claim 24, which is a therapeutic agent for vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.","26.A method of treating vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which comprises administrating a pharmaceutical composition of claims 24 or 25.27.Use of a polynucleotide having a base sequence as set forth in SEQ ID NO: 1 and a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 to produce a pharmaceutical composition of claim 24 or 25."],["<SOH> BACKGROUND ART <EOH>By a double adapter method, shotgun library of human genome was constructed and a nucleotide sequence of AF055021 (SEQ ID NO: 3), EST clone, was determined with the DNA library (ANLITICAL BIOCHEMISTRY, 236, 107-113 (1996).).","Subsequently, a nucleotide sequence of full-length cDNA of AF055021 (SEQ ID NO: 1) was determined and named KIAA1036 (DNA Research, 6 (3) 197-205 (1999).).","KIAA1036 polypeptide (SEQ ID NO: 2) is a polypeptide consisting of 365 amino acids and the function has not been elucidated yet.","Since cells need oxygen and nutriments provided from blood to survive in vivo, neovascularization is vigorous in vigorously propagating tissues.","It is known that neovascularization relates not only to the physiological regeneration of tissues but also to diseases accompanied by the pathological cell propagation.","As the disease, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease and cancer were reported (Nature, 407, 249 (2000).).","Since neovascularization is also observed upon the propagation of malignant neoplasma cell, human umbilical vein endothelial cells (HUVEC) treated with vascular endothelial cell growth factor (VEGF) were reported as a model of neovascularization caused by cancer (Cancer Research, 55, 5296-5301 (1995).).","Though the research of genes relating to the above-mentioned various diseases have been conducted, the mechanism of pathogenesis has not been perfectly elucidated, there being a possibility of unknown genes relating to the mechanism.","Under this circumstance, an identification of a new gene relating to the pathogenesis of the above-mentioned diseases and a diagnostic procedure of diseases with the gene have been desired."],["<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG.","1 is a schematic comparing an amino acid sequence of KIAA1036 and that of AK022567.Query: 614 shows an amino acid sequence of KIAA1036.Sbjct: 1 shows an amino acid sequence of AK022567.FIG.","2 shows an expression level of KIAA1036 in human umbilical vein endothelial cells by Northern blot.","Total RNA was prepared by treating human umbilical vein endothelial cell with nM VEGF for 0, 0.5, 2, 6, 12 or 24 hours.","And then signal intensity of KIAA1036 gene against signal intensity of β-actin gene was calculated.","A relative expression level is shown when signal intensity in a control division (1 nM VEGF was not added) is 1.The expression was inhibited in 0.5 to 2 hours after VEGF was added.","But a 3.2-fold high gene expression was induced at 24 hours after VEGF was added.","FIG.","3 shows an expression level of KIAA1036 in human cancer tissues by Dot blot.","By using a nylon membrane on which cDNA was immobilized, wherein cDNA was derived from a cancer tissue obtained from a patient with human ovarian cancer or large bowel cancer and a normal tissue obtained from the same patient, signal intensity of KIAA1036 gene against signal intensity of β-actin gene was calculated.","The ratio of signal intensity in each cancer tissue against in each normal tissue of the same patient was calculated.","And a change of expression level of the gene in each cancer tissue was analyzed.","As a result, in cancer tissue of all of the 4 patients with ovarian cancer and all of the 11 patients with large bowel cancer, the expression of KIAA1036 gene was increased.","FIG.","4 shows a cytostatic effect of KIAA1036 polypeptide against a vascular endothelial cell.","Cell propagation in human vascular endothelial cells was calculated by measuring an uptake of bromodeoxyuridine (BrdU) in DNA synthesis.","Human vascular endothelial cells were divided in M199 medium in which a concentration of KIAA1036 polypeptide was 0, 1 or 10 nM.","BrdU reagent was added thereto and the cells were cultured for 12 hours.","And then the quantity of BrdU taken in cells was measured (lane 1 to 3).","The same operation was carried out with M199 medium to which 1 nM VEGF was added and the quantity of BrdU taken in cells was measured (lane 4 to 6).","As a result, regardless of the addition of VEGF, DNA synthesis in cells was decreased, depending on the concentration of added KIAA1036 polypeptide.","FIG.","5 shows a depression effect of KIAA1036 against migrating activity of a vascular endothelial cell.","M199 medium, in which a concentration of KIAA1036 polypeptide was 0, 1 or 10 nM, was added in the lower layer of a culture apparatus divided with a filter and vascular endothelial cells were added in the upper layer of it.","The cells were cultured for 4 hours.","Then cell population attached to the filter was measured (lane 1 to 3).","The same operation was carried out with M199 medium to which 0.25 nM VEGF was added and cell population attached to the filter was measured (lane 4 to 6).","As a result, regardless of the addition of VEGF, migrating activity of cells was inhibited, depending on the concentration of added KIAA1036 polypeptide.","detailed-description description=\"Detailed Description\" end=\"lead\"?"],["TECHNICAL FIELD The present invention relates to a polypeptide and a polynucleotide useful as a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.","And the present invention relates to the treatment of those diseases.","BACKGROUND ART By a double adapter method, shotgun library of human genome was constructed and a nucleotide sequence of AF055021 (SEQ ID NO: 3), EST clone, was determined with the DNA library (ANLITICAL BIOCHEMISTRY, 236, 107-113 (1996).).","Subsequently, a nucleotide sequence of full-length cDNA of AF055021 (SEQ ID NO: 1) was determined and named KIAA1036 (DNA Research, 6 (3) 197-205 (1999).).","KIAA1036 polypeptide (SEQ ID NO: 2) is a polypeptide consisting of 365 amino acids and the function has not been elucidated yet.","Since cells need oxygen and nutriments provided from blood to survive in vivo, neovascularization is vigorous in vigorously propagating tissues.","It is known that neovascularization relates not only to the physiological regeneration of tissues but also to diseases accompanied by the pathological cell propagation.","As the disease, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease and cancer were reported (Nature, 407, 249 (2000).).","Since neovascularization is also observed upon the propagation of malignant neoplasma cell, human umbilical vein endothelial cells (HUVEC) treated with vascular endothelial cell growth factor (VEGF) were reported as a model of neovascularization caused by cancer (Cancer Research, 55, 5296-5301 (1995).).","Though the research of genes relating to the above-mentioned various diseases have been conducted, the mechanism of pathogenesis has not been perfectly elucidated, there being a possibility of unknown genes relating to the mechanism.","Under this circumstance, an identification of a new gene relating to the pathogenesis of the above-mentioned diseases and a diagnostic procedure of diseases with the gene have been desired.","DISCLOSURE OF INVENTION The present inventors found that expression of KIAA1036 polypeptide was increased in ovarian cancer and colon cancer and confirmed that the peptide was useful as a marker for cancer.","As results of further researches, they found that KIAA1036 polypeptide was a new marker for neovascular and useful as a marker and a therapeutic agent of various diseases such as vascular disease relating to neovascularization, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer to accomplish the present invention.","The present invention is (1) a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof, (2) a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polynucleotide hybridizing under a stringent condition with a polynucleotide having a base sequence as set forth in C of 1-C of 1480 of SEQ ID NO: 1 or fragment thereof, (3) a marker of (1), which contains a polynucleotide having a base sequence as set forth in C of 1-C of 1480 of SEQ ID NO: 1 or fragment thereof, (4) a marker of (1), which contains a polynucleotide having a base sequence as set forth in C of 1-C of 5481 of SEQ ID NO: 1 or fragment thereof, (5) a marker for neovascuarlization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (6) a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polypeptide wherein 1 or more amino acid residue(s) has a mutation(s) selected from deletion, substitution and addition in an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (7) a marker of (5), which contains a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (8) a method of detection of a marker of any one of (1)-(7) from a sample derived from human, (9) a method of detection or assay of a marker of any one of (1)-(4), which comprises the following processes and using a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof, (a) a process that the polynucleotide and a sample are contacted, and (b) a process that a polynucleotide binding with the polynucleotide is detected, (10) a method of detection or assay of a marker of any one of (1)-(4), which comprises the following processes and using a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof, (a) a process that cDNA is prepared from a sample, and (b) a process that cDNA is amplified with the polynucleotide fragment and detected, (11) a method of detection or assay of a marker of any one of (5)-(7), which comprises the following processes and using an antibody recognizing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (a) a process that the antibody and a sample are contacted, and (b) a process that the binding between the antibody and a polypeptide is detected, (12) a method of detection of a disease relating to a marker of any one of (1)-(7), which comprises using a method of any one of (9)-(11), (13) a method of detection of (12), wherein the disease is vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, (14) a diagnostic kit of a disease relating to a marker of any one of (1)-(7), which comprises using a method of any one of (9)-(11), (15) a diagnostic kit of (14), wherein the disease is vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, (16) a screening method of a binding substance of a marker of any one of (5)-(7), which comprises the following processes, (a) a process that a marker of any one of (5)-(7) and a subject are contacted, and (b) a process that the binding between the marker and the subject is detected, (17) a kit for screening of a binding substance of (16), which contains a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof, (18) a screening method of a binding activity regulatory substance which regulates a binding between a marker of any one of (5)-(7) and a binding substance of the marker, which comprises the following processes, (a) a process that a marker of any one of (5)-(7) and a binding substance of the marker are contacted in the presence or absence of a subject, and (b) a process that a binding activity of the marker and the binding substance is compared in the presence or absence of a subject, (19) a kit for screening of a binding activity regulatory substance of (18), which contains a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof, (20) a screening method of an expression regulatory substance of a marker of any one of (1)-(7), which comprises the following processes, (a) a process that a cell, which can express a marker of any one of (1)-(7), and a subject are contacted, and (b) a process that a change of an expression level of the marker is detected by using a method of any one of (9)-(11), (21) a kit for screening of an expression regulatory substance of a marker of any one of (1)-(7), which comprises using the method of (20), (22) a DNA synthesis inhibitor containing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (23) a cell migrating activity inhibitor containing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (24) a pharmaceutical composition which contains at least one selected from a DNA synthesis inhibitor of (22), a cell migrating activity inhibitor of (23), a polynucleotide having a base sequence as set forth in SEQ ID NO: 1 or fragment thereof, a human gene therapeutic expression vector containing polynucleotide having a base sequence as set forth in SEQ ID NO: 1 or fragment thereof, a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof and an antibody to the polypeptide, (25) a pharmaceutical composition of (24), which is a therapeutic agent for vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, (26) a method of treating vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which comprises administrating a pharmaceutical composition of (24) or (25), (27) use of a polynucleotide having a base sequence as set forth in SEQ ID NO: 1 and a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 to produce a pharmaceutical composition of (24) or (25).","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is a schematic comparing an amino acid sequence of KIAA1036 and that of AK022567.Query: 614 shows an amino acid sequence of KIAA1036.Sbjct: 1 shows an amino acid sequence of AK022567.FIG.","2 shows an expression level of KIAA1036 in human umbilical vein endothelial cells by Northern blot.","Total RNA was prepared by treating human umbilical vein endothelial cell with nM VEGF for 0, 0.5, 2, 6, 12 or 24 hours.","And then signal intensity of KIAA1036 gene against signal intensity of β-actin gene was calculated.","A relative expression level is shown when signal intensity in a control division (1 nM VEGF was not added) is 1.The expression was inhibited in 0.5 to 2 hours after VEGF was added.","But a 3.2-fold high gene expression was induced at 24 hours after VEGF was added.","FIG.","3 shows an expression level of KIAA1036 in human cancer tissues by Dot blot.","By using a nylon membrane on which cDNA was immobilized, wherein cDNA was derived from a cancer tissue obtained from a patient with human ovarian cancer or large bowel cancer and a normal tissue obtained from the same patient, signal intensity of KIAA1036 gene against signal intensity of β-actin gene was calculated.","The ratio of signal intensity in each cancer tissue against in each normal tissue of the same patient was calculated.","And a change of expression level of the gene in each cancer tissue was analyzed.","As a result, in cancer tissue of all of the 4 patients with ovarian cancer and all of the 11 patients with large bowel cancer, the expression of KIAA1036 gene was increased.","FIG.","4 shows a cytostatic effect of KIAA1036 polypeptide against a vascular endothelial cell.","Cell propagation in human vascular endothelial cells was calculated by measuring an uptake of bromodeoxyuridine (BrdU) in DNA synthesis.","Human vascular endothelial cells were divided in M199 medium in which a concentration of KIAA1036 polypeptide was 0, 1 or 10 nM.","BrdU reagent was added thereto and the cells were cultured for 12 hours.","And then the quantity of BrdU taken in cells was measured (lane 1 to 3).","The same operation was carried out with M199 medium to which 1 nM VEGF was added and the quantity of BrdU taken in cells was measured (lane 4 to 6).","As a result, regardless of the addition of VEGF, DNA synthesis in cells was decreased, depending on the concentration of added KIAA1036 polypeptide.","FIG.","5 shows a depression effect of KIAA1036 against migrating activity of a vascular endothelial cell.","M199 medium, in which a concentration of KIAA1036 polypeptide was 0, 1 or 10 nM, was added in the lower layer of a culture apparatus divided with a filter and vascular endothelial cells were added in the upper layer of it.","The cells were cultured for 4 hours.","Then cell population attached to the filter was measured (lane 1 to 3).","The same operation was carried out with M199 medium to which 0.25 nM VEGF was added and cell population attached to the filter was measured (lane 4 to 6).","As a result, regardless of the addition of VEGF, migrating activity of cells was inhibited, depending on the concentration of added KIAA1036 polypeptide.","BEST MODE FOR CARRYING OUT THE INVENTION Preparation of KIAA1036 polynucleotide which is a marker of the present invention, preparation of the polypeptide, a recombinant vector, a transformant, a method of production thereof, an antibody, the method of detection of polypeptide, a method of detection of mRNA, the method of detection of neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer and a diagnostic kit, a therapeutic agent or a method of treating for vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer are explained below.","In this description, if there is no instruction, a gene recombinant technique, a production engineering technique of a recombinant polypeptide in animal cells, insect cells, yeast and Escherichia coli, a molecular-biological method, the method of separation and purification of expressed polypeptide, assay and immunological method, which are well-known in this field, are adopted.","A marker of the present invention is “a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof” and a polynucleotide sequence of KIAA1036.Additionally, a marker of the present invention includes “a polynucleotide having a base sequence as set forth in C of 1-C of 1480 of SEQ ID NO: 1 or fragment thereof” and “a polynucleotide hybridizing with the present invention marker under a stringent condition or fragment thereof”.","Hereinafter, a polynucleotide, which is a marker of the present invention, is referred to as polynucleotide A.","In this description, “neovascularization” means a phenomenon that a new vasculature is developed from an existing vasculature.","A disease related to “neovascular” includes “vascular disease”, “inflammatory disease”, “entoptic neovascular disease”, “reproductive system disease”, “central nervous system disease” or “cancer”.","“Vascular disease” includes arterial sclerosis, hypertonia, angina pectoris, obstructive arteriosclerosis, myocardial infarction, cerebral infarction, diabetic angiopathy or vascular malformation.","“Inflammatory disease” includes hepatitis, pneumonitis, glomerular nephritis, thyreoiditis, osteitis, arthromeningitis, osteoclasia, chondrolysis, rheumatism, bronchial asthma, sarcoidosis, Crow-Fukase syndrome, pannus, allergic oedema, ulcers, hydroperitoneum, peritoneal screlosis or tissular conglutination.","“Entoptic neovascular disease” includes diabetic retinopathy, occlusion of retinal vein or aging macular degeneration.","“Reproductive system disease” includes uterus dysfunction, placental dysfunction, ovarian hyperergasia or follicle cyst.","“Central nervous system disease” includes retinosis, cerebral apoplexy, vascular dementia or Alzheimer disease.","“Cancer” includes a malignant neoplasma such as solid cancer, angiomatous, hemangioendothelioma, sarcomas, sarcoma or hematopoietic organic ulcer, ovarian cancer or large bowel cancer.","Additionally, it includes metastatics of these cancers.","“A polynucleotide hybridizing under a stringent condition” means a polynucleotide obtainable by a generally known and common method in this field with using a fragment of polynucleotide A as a probe, for example, colony hybridization, plaque hybridization or Southern blotting hybridization.","An example of the above polynucleotide includes a polynucleotide obtained by hybridization at 65° C. in the presence of 0.7 to 1.0M NaCl by using a membrane on which polynucleotides from colony or plaque are immobilized and a washing of a membrane at 65° C. with SSC (Saline Sodium Citrate; 150 mM sodium chloride, 15 mM sodium citrate) solution whose concentration is 0.1-fold to twice.","Hybridization can be performed as well as methods described in Molecular Cloning: A Laboratory Manual, Second Edition (1989)(Cold Spring Harbor Laboratory Press), Current Protocols in Molecular Biology (1994)(Wiley-Interscience), DNA Cloning 1: Core Techniques, A practical Approach, Second Edition (1995)(Oxford University Press).","Preferably, sequences comprising of only adenine (a) or thymine (T) are excluded from sequences hybridizing under a stringent condition.","“A fragment of a polynucleotide” includes a polynucleotide having a base sequence comprising 5 or more serial bases in a base sequence of SEQ ID NO: 1, for example, a polynucleotide having a base sequence comprising 5, 8, 10, 12, 15, 20, 25, 30, 50, 100, 500 or 1000 bases.","In this description, “a polynucleotide hybridizing” includes a polynucleotide hybridizing with the other polynucleotide under the above hybridizing condition.","Examples of the above polynucleotide includes a polynucleotide having a homology of at least 60% or more, preferably 80% or more and more preferably 95% or more with a base sequence of DNA encoding a polypeptide having an amino acid sequence of SEQ ID NO: 1.And the homology is shown as a score, for example, by using a search program BLAST with algorithm developing by Altschul et al (The Journal of Molecular Biology, 215, 403-410 (1990).).","A marker of the present invention includes “a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof”, a polypeptide sequence of KIAA1036 including their salts.","A “salt” includes a salt physiologically acceptable with an acid or a base.","A salt with an acid is especially preferable.","The salt includes, for example, a salt with inorganic acid such as hydrochloric acid, phosphoric acid, hydrobromic acid or sulfuric acid, or a salt with organic acid such as acetic acid, fomic acid, proprionic acid, fumaric acid, maleic acid, succinic acid, tartaric acid, citric acid, malic acid, oxalic acid, benzoic acid, methanesulfonic acid or benzenesulfonic acid.","“A fragment of polypeptide” includes a polypeptide sequence comprising 5 or more serial amino acid residues in an amino acid sequence of SEQ ID NO: 2, for example, a polypeptide comprising an amino acid sequences comprising 5, 8, 10, 12, 15, 20, 25, 30, 50 or 100 amino acid residues.","A marker of the present invention includes “a polypeptide having a mutation selected from deletion, substitution and addition in 1 or a few amino acid residue(s) of an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof”.","Hereinafter, a polypeptide, which is a marker of the present invention, is referred to as polypeptide B.","“1 or a few amino acid residue(s)” means amino acid residues, which can delete, substitute or add by a site-specific mutagenic method or the like.","Namely, it means amino acid residues having 50 or less, preferably 30 or less, more preferably 20 or less, much more preferably 10 or less amino acid residues.","Furthermore, polypeptide B includes a polypeptide to be used as a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer even after deletions, substitutions or additions.","It is, for example, a polypeptide having a homology of at least 60% or more, preferably 80% or more and more preferably 95% or more with an amino acid sequence of SEQ ID NO: 2.In this description, “a marker” includes a substance used to detect neovascularization and all kinds of disease related to neovascularization derived from organism samples, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.","And it includes polynucleotide A and polypeptide B. mRNA or a protein can be a “marker” when the expression increases or decreases under diseases.","Examples of the above maker include KIAA1036 polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2, KIAA1036 polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1, a polynucleotide having a complementary base sequence thereof or a fragment thereof.","In this description, terms are used with usual meanings in this field unless otherwise mentioned especially referred.","The terms especially used in this description are explained below.","An “antibody” means a general antibody in this field and includes all of the antibodies, fragments thereof, derivatives, conjugations or modified antibodies.","An antibody recognizing polypeptide B or fragment thereof is preferable, an antibody recognizing specifically the above polypeptide is more preferable and an antibody recognizing single specifically the polypeptide is much more preferable.","These antibodies include a polyclonal antibody or a monoclonal antibody.","“A detection or a quantitation” of a marker of the present invention can be accomplished by any appropriate method including an immunological assay or a molecular-biological assay.","When a marker is polypeptide B, the above method includes, for example, an immunological assay such as ELISA (Enzyme Linked Immuno Sorbent Assay), RIA (Radio Immuno Assay), fluorescence antibody technique, Western blot or an immune structure dyeing method.","By using an antibody against polypeptide B or fragment thereof, neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer can be detected or quantitated.","Because a marker of the present invention can be used as a marker for above diseases, a method of detection or quantitation of diseases with the above antibody can be accomplished by any appropriate method including an immunological assay as well as an above detection or quantitation of a polypeptide.","The present invention relates to a diagnostic kit of neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, characterized by including an antibody against polypeptide B or fragment thereof.","The above kit includes at least an antibody against polypeptide B or fragment thereof and a standard reagent of polypeptide B.","Because polypeptide B can be used as a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, it can be immunologically measured as well as an above mentioned method of detection or quantitation by an immunological assay.","And when a marker is a polynucleotide such as mRNA, the assay includes a molecular-biological assay, for example, Northern blot, Dot blot or polymerase chain reaction (PCR).","mRNA can be detected or quantitated by using polynucleotide A or fragment thereof as a probe or primer.","Because a marker of the present invention can be a marker of above diseases, a method of detection or quantitation of diseases with the probe or the primer is accomplished by an appropriate method including a molecular-biological assay as well as the above method for detection or quantitation of a polynucleotide.","A probe or a primer includes a nucleotide comprising serial bases in a base sequence of SEQ ID NO: 1, for example, a base sequence comprising 5, 8, 10, 12, 15, 20, 25, 30, 40, 50 or 60 bases.","It includes an oligonucleotide having the same sequence, an oligonucleotide having a sequence complementary to the sequence of the oligonucleotide, and the derivative thereof.","A derivative thereof includes, for example, a oligonucleotide wherein a phosphodiester bond in the oligonucleotide is transformed into a phosphorothioate bond or a N3′-P5′ phosphoamidite bond, a oligonucleotide wherein a ribose and a phosphodiester bond are transformed into a peptide bond, a oligonucleotide wherein a uracil in the oligonucleotide is substituted with a C-5 propionyl uracil or a C-5 thiazole uracil, a oligonucleotide wherein a cytosine in the oligonucleotide is substituted with C-5 propionyl cytosine or cytosine modified with phenoxazine or a oligonucleotide wherein a ribose in DNA is substituted with 2′-O-propyl ribose, 2′-methoxyethoxy ribose or the like.","Those polynucleotides are useful, for example, as a gene marker, a primer for PCR or a probe for hybridization.","The present invention relates to a part or all of the polynucleotide encoding polypeptide B and a diagnostic kit of vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer including the polynucleotide as a standard reagent.","This kit includes at least a probe or a primer against polynucleotide A or fragment thereof and a standard reagent of polynucleotide A.","Because polynucleotide A can be used as a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, it is molecular-biologically measured as well as the above method of detection or quantitation by a molecular-biological assay.","A transcription of DNA or a translation of mRNA can be inhibited by using polynucleotide A.","Methods to inhibit transcription of DNA or translation of mRNA of the polypeptide B can be provided by appropriately preparing a polynucleotide having a complementary base sequence to a base sequence of polynucleotide A, a polynucleotide hybridizing with polynucleotide A or an oligonucleotide and a derivative oligonucleotide having a complementary sequence to an oligonucleotide having serial 5 to 60 base sequences in base sequence of polynucleotide A and using antisense RNA/DNA techniques known in this field.","The present invention relates to “a screening method of a binding substance” of a marker of the present invention.","“A binding substance” includes any substance binding to a marker of the present invention.","The binding substance includes, for example, a low-molecular weight substance, a polynucleotide or a polypeptide such as a receptor and an antibody.","“A screening method of a binding substance” can be accomplished by appropriately using techniques widely known in this technical field.","It includes, for example, a bioassay and a binding assay.","“A kit for screening of a binding substance” of the present invention includes at least a marker of the present invention.","A binding substance can be screened by biochemical method with the above kit.","Additionally the present invention relates to “a screening method of a binding activity regulatory substance”.","“A binding activity regulation” means that a binding activity of a marker of the present invention and the binding substance is enhanced or reduced and “a binding activity” is evaluated by Percent Maximum Binding (PMB).","“A binding activity regulatory substance” includes a substance that makes a binding activity of the binding substance against a marker of the present invention enhanced or reduced, for example, an agonist or an antagonist.","The substance includes, for example, a low-molecular weight substance, a polynucleotide or a polypeptide.","“A screening method of a binding activity regulatory substance” can be accomplished by appropriately using techniques widely known in this field.","A binding activity regulatory substance can be screened by a screening method of the present invention.","The method is, for example, a bioassay or a binding assay.","“A screening kit of a binding activity regulatory substance” of the present invention includes at least a marker of the present invention.","A binding activity regulatory substance can be screened by a biochemical method with a kit of the present invention.","And the present invention relates to “a screening method of an expression regulatory substance”.","Expression level means the amount of a marker of the present invention expressed in target cells.","Expression level can be evaluated by an appropriate method including an immunological assay with an antibody against polypeptide B, for example, ELISA, RIA, a fluorescence antibody technique or an immune structure dyeing method or by a molecular-biological assay measuring mRNA of polynucleotide A, for example, Northern blot hybridization, Dot blot or RT-PCR.","“Expression regulation” means that expression level of a marker of the present invention in protein or mRNA level evaluated by any appropriate method including the above immunological or molecular-biological assay is enhanced or reduced.","“An expression regulatory substance” includes a substance enhancing or reducing expression level of a marker of the present invention in protein or mRNA level, which can be evaluated by any appropriate method including the above immunological or molecular-biological assay.","The above substance includes, for example, a low-molecular weight substance, a polynucleotide or a polypeptide.","“A screening method of an expression regulatory substance” can be accomplished by appropriately using techniques widely known in this field.","An expression regulatory substance can be screened by a screening method of the present invention.","The method includes, for example, a bioassay or a binding assay.","“A screening kit of an expression regulatory substance” of the present invention includes at least a marker of the present invention.","An expression regulatory substance can be screened by a biochemical method based on a screening method of an expression regulatory substance with a kit of the present invention.","Because polypeptide B inhibits a DNA synthesis in a vascular endothelial cell and a migrating activity of a vascular endothelial cell, polynucleotide A, a complementary polynucleotide of the above polynucleotide, polypeptide B or/and an antibody against the above polypeptide can be a DNA synthesis inhibitor or/and a cell migrating activity inhibitor.","Then, the present invention provides a DNA synthesis inhibitor and a cell migrating inhibitor.","“A pharmaceutical composition” of the present invention includes at least one selected from polypeptide B, an antibody against the polypeptide, polynucleotide A, a complementary polynucleotide of the polynucleotide, a DNA synthesis inhibitor, a cell migrating activity inhibitor, a binding substance, a binding activity regulatory substance and an expression regulatory substance.","It can be used for a therapeutic agent of a specific disease, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.","The substance includes, for example, a low-molecular weight substance, a polynucleotide or a polypeptide.","Preferred is a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.","And a pharmaceutical composition of the present invention includes “a gene therapeutic expression vector”.","“A gene therapeutic expression vector” includes an expression vector inserted all or a part of polynucleotide A and a vector which inserts a normal gene in a cell, repairs/modifies a defect of a gene and regulates an expression of a gene by introducing a gene in a cell/tissue.","The vector includes a viral vector wherein a part or all of a base sequence of a viral, which is lack of reproductive capability, is replaced with a therapeutic gene.","Then, the present invention provides therapeutic agents or methods of treating for these diseases.","“A method of treating” includes a method for treating of specific diseases, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer relating to neovascularization by a administration of the above pharmaceutical composition.","Hereinafter, the Present Invention is Described in Detail.","(1) An Preparation of KIAA1036 Polynucleotide cDNA library is manufactured from human brain, heart, skeletal muscle, spleen, kidney, liver, small intestine, placenta, human normal cells from these tissues, human umbilical vein endothelial cells or human ovarian cancer or human large bowel cancer by a usual method.","A method which manufactures cDNA library is a method described in Molecular Cloning: A Laboratory Manual, Second Edition (1989) (Cold Spring Harbor Laboratory Press), Current Protocols in Molecular Biology (1994) (Wiley-Interscience) or DNA Cloning 1: Core Techniques, A Practical Approach, Second Edition (1995) (Oxford University Press) or the method that a kit on the market, for example, SuperScript Plasmid System for cDNA Synthesis and Plasmid Cloning (Invitrogen) or ZAP-cDNA Synthesis Kits (STRATAGENE) is used.","cDNA obtained by the method is, for example, DNA encoding a polypeptide of SEQ ID NO: 2.Examples of the above cDNA include a DNA comprising a base sequence of SEQ ID NO: 1.A plasmid including DNA of SEQ ID NO: 1 is, for example, a plasmid described in the following examples.","The obtained DNA is inserted to an appropriate expression vector and an expression plasmid is constructed.","The expression vectors are introduced to appropriate hosts and then transformants can be obtained.","The expression vector is any vector in which cDNA is inserted and which express in animal cells.","The vector is, for example, pcDNA1.1, pcDNA1.1/Amp, pCDM8, pREP (Invitrogen), pHM6, pHB6 (Roche Diagnostics), pKK223-3, pGEX (Amersham Pharmacia Bioteque), pET-3, pET-11, pBluescriptII SK(+), pBluescriptII SK(−) (STRATAGENE), pUC19, pTrxFus (Invitrogen), pUC118, pSTV28 (TaKaRa), pMAL-c2X (New England BioLabs), pAGE107 (Cytotechnology, 3 (2), 133-140 (1990).","; JP1991-22979), pAGE103 (The Journal of Biochemistry, 101 (5), 1307-1310 (1987).","), pAMo, pAMoA (The Journal of Biological Chemistry, 268 (30), 22782-22787 (1993).","), pAMoPRSA (JP1993-336963) or pAS3-3 (JP1990-227075).","The expression vectors with a cDNA insert introduce a target cDNA into optional animal cells and transformed cells are obtained.","Any method for introducing DNA can be used as a method for introducing the expression vectors a host.","When a host is animal cells, it is, for example, an electroporation (Cytotechnology, 3 (2), 133-140 (1990).","), a calcium phosphate method (JP1990-227075) or a lipofection method (Proceedings of the National Academy of Sciences USA, 84, 7413 (1987).","; Virology, 52, 456 (1973).).","Appropriate cells or tissue for expression vectors can be used as a host.","It is, for example, animal cells.","An animal cell as a host is, for example, Namalwa (Burkitt lymphoma, ATCC: CRL-1432) and the subline NamalwaKJM-1 which is an established cell from human, HCT-15 (human large bowel cancer cell, ATCC: CCL-225), COS-1(African green monkey's nephrocyte (SV40 transformed cell), ATCC: CRL-1650) and COS-7 (African green monkey's nephrocyte (SV40 transformed cell), ATCC: CRL-1651) which is an established cell from monkey or CHO-K1 (Chinese hamster ovary cell, ATCC: CCL-61) and HBT5637 (JP1987-299) which is an established cell derived from hamster.","Preferred is Namalwa cell, NamalwaKJM-1 cell or HCT-15 cell.","Transformants of the present invention are cultured by a generally known and common method in this field.","It can be accomplished with a medium appropriate to a transforming host and a liquid medium is appropriate as a medium for a culturing liquid medium.","For example, MEM medium (Science, 130, 432 (1959).","), D-MEM medium (Virology, 8, 396 (1959).","), PRMI 1640 medium (The Journal of the American Medical Association, 199, 519 (1967).","), YT medium or BEM medium can be used.","When transformants is prepared by using animal cells as a host are cultured, for example, a medium wherein cattle fetal serum (FCS) is appropriately added into MEM medium, D-MEM medium or PRIM medium can be used.","A medium can optionally include a substance promoting transcription activity to enhance transcription activity of a promoter of an expression vector.","For example, isopropyl-1-thio-β-D-galactopyranosin (IPTG) can be used.","A medium includes nutriments needed to grow up transformants, for example, glucose, amino acid, peptone, vitamin, hormone or serum, preferably, FCS, calcium chloride or magnesium chloride.","The medium like this, which has any composition, can be used and a medium available on the market can be used.","A cultivation is held in pH 6.0 to 8.0 at 25 to 40° C. in the presence of 5% CO2.The obtained transformed cells are cultured by a usual method.","For example, they can be cultured by the following culture method.","When transformants are animal cells, examples of a medium for culturing the cells includes commonly used RPMI1640 medium (The Journal of the American Medical Association, 199, 519 (1967).","), MEM medium of Eagle (Science, 122, 501 (1952).","), D-MEM medium (Virology, 8, 396 (1959).","), 199 medium (Proceeding of the Society for the Biological Medicine, 73, 1 (1950).)","or a medium added fetal calf serum (FCS).","Cultivation is usually carried out in pH 6 to 8, at 25 to 40° C., in the presence of 5% CO2 for 1 to 7 days.","An antibiotic such as kanamycin, penicillin or streptomycin can optionally be added during cultivation.","DNAs encoding KIAA1036 which is a neovascular marker can be obtained from human brain, heart, skeletal muscle, spleen, kidney, liver, lung, placenta, human normal cells from these tissues, human umbilical vein endothelial cells or human ovarian cancer or human large bowel cancer by the above-mentioned.","On the basis of an amino acid sequence, DNA encoding KIAA1036 can be prepared by a chemical synthesis.","A chemical synthesis of DNA can be accomplished with a DNA synthesizer using a thiophosphite method (Shimazu Corporation) or a DNA synthesizer model 392 using a phosphoamidite method (PerkinElmer, Inc.).","The following oligonucleotide is used as a sense primer (SEQ ID NO: 4) or an antisense primer (SEQ ID NO: 5) and cDNA prepared from mRNA in cells expressing complementary mRNA of these DNA is used as a template.","Target DNA can be prepared by PCR under this condition.","(2) The Production of KIAA1036 Polypeptide KIAA1036 can be produced by a method described in Molecular Cloning: A Laboratory Manual, Second Edition (1989)(Cold Spring Harbor Laboratory Press) or Current Protocols in Molecular Biology (1994)(Wiley-InterScience).","For example, KIAA1036 polynucleotide is made to express in host cells by the following method and KIAA1036 can be produced.","On a basis of a full length of DNA encoding KIAA1036 polypeptide, an appropriate length of DNA fragment including a part encoding the polypeptide is optionally prepared.","And DNA whose bases substituted is prepared, as a base sequence encoding the polypeptide is the most suitable codons for expression in a host.","This DNA is useful to enhance productivity of the polypeptide.","By inserting the DNA fragment or a full length of DNA to the downstream of a promoter of an appropriate expression vector, a recombinant DNA (a recombinant vector) is produced.","By the recombinant vectors are produced in host cells adapting to the expression vector, transformants producing KIAA1036 polypeptide can be obtained.","Any cell such as a prokaryotic cell, yeast, an animal cell, a plant cell or a insect cell can be used as a host cell, if a target gene can be expressed in the cells.","As an expression vector, the vector which can autonomously replicate in the above host cells or be inserted into a chromosome and which includes a promoter at the appreciate position for the transcription of KIAA1036 polypeptide gene.","(i) The case that prokaryote is used as a host.","An expression vector of KIAA1036 can autonomously replicate in prokaryote as well as is preferably constructed with a promoter, a ribosome binding sequence, KIAA1036 and a transcription termination sequence.","A gene regulating a promoter can be included.","An expression vector is, for example, pBTrp2, pBTac1, pBTac2 (Roche Diagnostics), BluescriptII SK(+), pBluescriptII SK(−) (STRATAGENE), pSTV28, pUC118, pUC19 (TaKaRa), pKK233-2 (Pharmacia), pSE280, pSupex, pUB110, pTP5, pC194, pTrxFus (Invitrogen), pGEMEX-1 (Promega), pQE-8 (QIAGEN), pGEX (Pharmacia), pETsystem (Novagen), pMAL-c2 (New England BioLabs), pKYP10 (JP1982-110600), pKYP200 (Agricultural Biological Chemistry, 48, 669 (1984).","), pLSA1 (Agricultural Biological Chemistry, 53, 277 (1989).","), pGEL1 (Proceedings of the National Academy of Sciences USA, 82, 4306(1985).","), pEG400 (Journal of Bacteriology, 172, 2392(1990).","), pTrs30 (FERM BP-5407), pTrs32 (FERM BP-5408), pGHA2 (FERM BP-400), pGKA2 (FERM BP-6798), pPA1 (JP1987-233798) or pTerm2 (JP1990-22979, U.S. Pat.","No.","4,686,191, U.S. Pat.","No.","4,939,094, U.S. Pat.","No.","5,160,735).","Any promoter which can express in a host cell such as Escherichia coli can be used.","For example, it is a promoter from Escherichia coli or a phage such as trp promoter (Ptrp), lac promoter (Plac), PL promoter, PR promoter or PSE promoter, SPO1 promoter, SPO2 promoter or penP promoter.","And a promoter artificially modified such as a promoter which two Ptrps is connected in series (Ptrp×2), tac promoter, lacT7 promoter or letI promoter can be used.","Preferred is a plasmid that the distance between Shine-Dalgarno sequence which is a ribosome binding sequence and an initiation codon is regulated to an appropriate distance, for example, 6 to 18 bases.","A transcription termination sequence is not always needed for an expression of KIAA1036 polynucleotide but preferred is that a transcription termination sequence is positioned at the right downstream of a structural gene.","A host cell is, for example, a prokaryote of Escherichia genus, Serratia genus, Bacillus genus, Brevibacterium genus, Corynebacterium genus, Microbacterium genus or Pseudomonas genus, for example, XL1-Blue strain, XL2-Blue strain, DH1 strain, MC1000 strain, KY3276 strain, W1485 strain, JM109 strain, HB101 strain, No.","49 strain, W3110 strain, NY49 strain, BL21 (DE3) strain, BL21 (DE3) pLysS strain, HMS174 (DE3) strain or HMS174 (DE3) pLysS strain of E. coli as Escherichia genus, S. ficaria strain, S. fonticola strain, S. liquefaciens strain or S. marcescens strain as Serratia genus, B. subtilis strain or B. amyloliquefaciens strain as Bacillus genus, B. ammoniagenes strain, B. Immariophilum (ATCC: 14068) strain or B. saccharolyticum (ATCC: 14066) strain as Brevibacterium genus, C. glutamicum (ATCC: 13032) strain, C. glutamicum (ATCC: 14067) strain, C. glutamicum (ATCC: 13869) strain or C. acetoacidophilum (ATCC: 13870) strain as Corynebacterium genus, M. ammoniaphilum (ATCC: 15354) strain as Microbacterium genus or S. mephitica strain as Pseudomonas genus.","Any method to introduce DNA to the above host cell can be used as an introduction method of a recombinant vector, for example, an electroporation (Nucleic Acids Research, 16, 6127 (1988).","), calcium phosphate method (Proceedings of the National Academy of Sciences USA, 69, 2110 (1972).","), a protoplast method (JP1987-2483942) or the method described in Gene, 17, 107 (1982), or Molecular & General Genetics, 168, 111 (1979).","(ii) The Case that Yeast is Used as a Host.","When yeast is used as a host, an expression vector is, for example, YEp13 (ATCC:37115), YEp24 (ATCC:37051), YCp50 (ATCC:37419), pHS19 or pHS15.Any promoter which express in yeast can be used as a promoter, for example, ADH1 (alcohol dehydrogenase) promoter, PHO5 (acid phosphatase) promoter, PGK1 (phosphoglycerate kinase) promoter, GAPDH (glyceraldehyde3-phosphate dehydrogenase) promoter, GAL1 (galactose kinase) promoter, GAL10 (UDP galactose 4-epimerase) promoter, MF α 1 (α pheromone) promoter or CUP1 (metallothionein) promoter.","A host is, for example, S. cerevisiae species of Saccharomyces genus, S. pombe species of Schizosaccharomyces genus, K. lactis species of Kluyveromyces genus, T. pullulans species of Trichosporon genus, S. alluvius species of Schwanniomyces genus or P. pastoris species of Pichia genus.","Any method to introduce DNA into a host can be used as a introduction method of a recombinant vector, for example, an electroporation (Methods in Enzymology, 194, 182 (1990).","), a spheroplast method (Proceedings of the National Academy of Sciences USA, 84, 1929 (1978).)","or a lithium acetate method (Journal of Bacteriology, 153, 163 (1983), or Proceedings of the National Academy of Sciences USA, 75, 1929 (1978).).","(iii) The Case that an Animal Cell is Used as a Host.","When an animal cell is used as a host, an expression vector is, for example, pcDNA1/Amp, pcDNA1, pCDM8, pREP4 (Invitrogen), pAGE 107 (Cytotechnology, 3, 133 (1990).","), pAGE 103 (The Journal of Biochemistry, 101, 1307 (1987).","), pAMo, pAMoA (pAMoPRSA) (The Journal of Biological Chemistry, 268, 22782-22787 (1993).)","or pAS3-3 (JP1990-22705).","Any promoter that can express in a host can be used as a promoter, for example, a promoter of IE (Imediate-early) gene of human cytomegalovirus (hCMV), an early promoter of SV40, Long Terminal Repeat Promoter of Moloney Murine Leulemia Virus, a promoter of retrovirus, HSP promoter, SR α promoter or a promoter of metallothionein.","An enhancer of IE gene of human CMV can be used with a promoter.","An animal cell as a host is, for example, HEK293 (a human fetal nephrocyte, ATCC:CRL-1573), Namalwa (Burkitt lymphoma, ATCC:CRL-1432), HeLa (a cell of carcinoma of uterine cervix, ATCC:CCL-2), HBT5637 (a leukemia cell, JP1987-299), BALL-1 (a leukemia cell) or HCT-15 (a large bowel cancer cell) of an established cell from a human, Sp2/0-Ag14 (a mouse myeloma cell, ATCC:CRL-1581) or NSO (a mouse myeloma cell) of an established cell from a mouse, COS-1 (African green monkey nephrocyte (SV40 transformed cell), ATCC:CRL-1650) or COS-7 (African green monkey nephrocyte (SV40 transformed cell), ATCC:CRL-1651) of an established cell from a monkey, CHO-K1 (Chinese hamster ovary cell, ATCC:CCL-61) or BHK-21 (C-13) (Sicilian hamster kidney cell, ATCC:CCL-10) of an established cell from a hamster, PC12 (an adrenal pheochromocytoma, ATCC:CRL-1721) or YB2/0 (a rat myeloma cell, ATCC:CRL-1662) of an established cell from a rat.","Any method to introduce DNA into a host can be used as an introduction method of a recombinant vector, for example, an electroporation (Cytotechnology, 3, 133, (1990).","), a calcium phosphate method (JP1990-22705) or a lipofection method (Proceedings of the National Academy of Sciences, USA, 84, 7413 (1987), or Virology, 52, 456 (1973).).","(iv) The Case that a Plant Cell is Used as a Host.","When a plant cell or a plant is used as a host, a polypeptide can be produced as well as a well-known method (The Tissue Culture, 20 (1994)., The Tissue Culture, 21 (1995), or Trends in Biotechnology, 15, 45 (1997).).","An expression vector is, for example, Ti plasmid or Tobacco mosaic virus vector.","Any promoter that can express in a plant cell can be used as a promoter for a gene expression, for example, 35S promoter of Cauliflower mosaic virus (CaMV) or Rice actin 1 promoter.","And expression productivity of a gene can be enhanced by inserting intron 1 of an alcohol dehydrogenase gene of maize between a promoter and an expressed.","A host is, for example, a plant cell such as potato, tobacco, maize, rice, rape, soybean, tomato, carrot, wheat, barley, rye, alfalfa or flax.","Any method to introduce DNA into a host can be used as a introduction method of a recombinant vector, for example, a method with Agrobacterium (JP1983-140885, JP1984-70080 or WO94/00977), an electroporation (JP1984-251887) or a particle gun (gene gun) method (JP2606856, JP2517813).","(v) The Case that an Insect Cell is Used as a Host.","When an insect cell is used as a host, a transfer vector is, for example, pVL1392, pVL1393 or pBlueBacIII (Invitrogen) and a virus for infection is, for example, a Vaculovirus which infects insects of Mamestra brassicoe family; Autographa california nuclear polyhedrosis virus (AcMNPV) Bac-N-Blue DNA.","A transformation method of an insect cell is, for example, a method described in Baculovirus Expression Vector: A Laboratory Manual (1992) (W.H.","Freeman and Company), Molecular Cloning: A Laboratory Manual, Second Edition (1989) (Cold Spring Harbor Laboratory Press), Current Protocols in Molecular Biology (1994) (Wiley-InterScience) or Biotechnology, 6, 47 (1988).","A transfer vector including a target gene and baculovirus DNA for infection to an insect cell are added into a culture and a virus expressing a target gene produced by recombinant infects an insect cell to be expressed a polypeptide.","An insect cell as a host is, for example, an established cell from Spodoptera frugiperda (Mamestra brassicoe) or an established cell from Trichoplusia ni.","For example, a cell from S. frugiperda includes Sf9 (ATCC: CRL-1711, an ovary cell) or Sf21 (an ovary cell) and a cell strain from T.ni is, for example, High Five or BTI-TN-5B1-4 (an egg cell, Invitrogen).","Any method to introduce into a host can be used as an introduction method of a recombinant vector, for example, a calcium phosphate method (JP1990-22705) or a lipofection method (Proceedings of the National Academy of Sciences USA, 84, 7413 (1987).).","And an electroporation (Cytotechnology, 3, 133 (1990).)","can be used as well as an animal cell.","(vi) A Culture Method When a transformant having a recombinant vector with an inserted DNA encoding KIAA1036 polypeptide of the present invention is a cell such as Escherichia coli, yeast, an animal cell or a plant cell, a cultivation by a usual culture method suited to all kinds of hosts is held.","And the polypeptides are produced, accumulated and collected from transformants or a culture solution to produce the polypeptide.","When transformants are animals or plants, they are cultured by a usual growth method suited to all kinds of hosts.","And the polypeptides are produced, accumulated and collected from animals or plants to produce the polypeptide.","When a host is an animal, for example, a nonhuman transgenic animal having polynucleotides of the present invention is cultured.","And KIAA1036 polypeptides encoded by the recombinant DNA are produced and accumulated in the animal and they are collected from an animal to produce KIAA1036 polypeptides.","A production/accumulation place in an animal is, for example, milk, sputum or egg of the animal.","When a host is a plant, for example, a transgenic plant having KIAA1036 polynucleotides of the present invention is cultured.","And KIAA1036 polypeptides encoded by the recombinant DNA are produced or accumulated in a plant and collected from a plant to produce KIAA1036 polypeptides.","When a host is a prokaryote such as Escherichia coli or a eucaryote such as yeast, for example, transformants having polynucleotides of the present invention are cultured in a medium.","And KIAA1036 polypeptides encoded by the recombinant DNA are produced or accumulated in a culture solution and collected from the culture to produce polypeptides of the present invention.","The method that transformants of KIAA1036 of the present invention are cultured in a medium is accomplished by a usual method for cultivation.","When a host is prokaryote such as Escherichia coli or a eucaryote, a natural medium or a synthetic medium can be used as a medium that obtained transformants are cultured if it has a carbon source, a nitrogen source and mineral that a host can assimilate and it is a medium that cultivation of transformants is held efficiently.","For example, YT medium including bactotryptone, yeast extract and sodium chloride is preferable as a medium when transformants that hosts are Escherichia coli are cultured.","Any carbon source that each microorganism can assimilate can be used, for example, glucose, fructose, sucrose, syrup including them, carbohydrate such as starch or starch hydrolysate, an organic acid such as acetic acid or proprionic acid or alcohol such as ethanol or propanol.","A nitrogen source is, for example, all kinds of inorganic acids such ammonia, ammonium chloride, ammonium sulfate, ammonium acetate, ammonium phosphate, ammonium salts of organic acids, other nitrogenous substances, peptone, meat extract, yeast extract, Corn Steep Liquor, casein hydrolysate, soybean cake, soybean cake hydrolysate, all kinds of fermentative bacteria or the digest.","A mineral is, for example, potassium phosphate monobasic, potassium phosphate dibasic, magnesium phosphate, magnesium sulfate, sodium chloride, ferrous sulfate, manganese sulfate, copper sulfate or calcium carbonate.","Cultivation is held under an aerobic condition such as a shaking culture or a submerged culture.","An incubation temperature is preferably 15 to 40° C. and a culture time is usually 5 hours to 7 days.","During cultivation, pH is kept from 3.0 to 9.0.Adjustment of pH is held with mineral or organic acid, alkali solution, urea, calcium carbonate or ammonia.","And an antibiotic such as ampicillin or tetracycline can be optionally added into a medium during cultivation.","When microorganisms transforming with an expression vector with an inductive promoter are cultured, inducers can be optionally added into a medium.","For example, when transformants transforming with an expression vector with lac promoter are cultured, isopropyl β-D-thiogalactopyranoside can be added into a medium.","When transformants transforming with an expression vector with trp promoter are cultured, indoleacrylic acid can be added into a medium.","Cells and organs of plants introduced genes can do a mass culture with jar fermenter.","A culture medium is, for example, popularly used Murashige & Skoog (MS) medium, White medium, or a medium that plant hormones such as oxine or cytokinin are added into these mediums.","When transformants for production of KIAA1036 polypeptide are animal cells, a medium that cells are cultured is popularly used RPMI1640 medium (The Journal of the American Medical Association, 199, 519 (1967).","), MEM medium (Science, 130, 432 (1959).","), D-MEM medium (Virology, 8, 396 (1959).","), 199 medium (Proceedings of the Society for the Biological Medicine, 73, 1 (1950).)","or a medium that fetal calf serum (FCS) is added into these mediums.","Cultivation held usually in pH 6 to 8, at 25 to 40° C., in the presence of 5% CO2 for 1 to 7 days.","And an antibiotic such as kanamycin, penicillin or streptomycin can be optionally added into a medium during cultivation.","When transformants are insect cells, a culture medium is popularly used TNM-FH medium (Pharmingen), Sf-900II SFM medium (Invitrogen), ExCell400, ExCell405 (JRH Biosciences Inc.) or Grace's Insect Medium (Nature, 195, 788 (1962).).","(vii) The Method of Production Transformants are cultured and KIAA1036 polypeptides are isolated and purified from a culture to produce KIAA1036 polypeptides.","A method of isolation/purification of KIAA1036 polypeptides can be usual method widely known in this field, for example, a method of isolation/purification of an enzyme or a method of purification of transglucosylase by Sandler (Methods in Enzymology, 83, 458).","When KIAA1036 polypeptides are produced and accumulated as dissolved polypeptides, a culture solution that transformants are cultured as mentioned above is separated to cells or fungus body and a medium, for example, by a centrifugal separation.","When KIAA1036 polypeptides exist in host cells, after cells or fungus bodies extracted are washed with an appropriate buffer such as STE solution and broken into pieces by ultrasonic waves, French press, Manton Gaulin homogenizer or Dynomill, KIAA polypeptides can be obtained as an acellular solution by a centrifugal separation or a filtration.","The suitable quantity of surfactant can be included in a buffer for separation/purification of KIAA1036 polypeptide.","For example, sodium lauryl sulfate (SDS) or Sodium N-Dodecanoylsalcosinate (salcosiyl) can be used.","A method of separation/purification of target proteins included in an obtained crude material can be accomplished with the combination of all kinds of well-known methods of separation/purification.","The well-known method is, for example, a solvent extraction method, a salting-out method with ammonium sulfate, a dialysis, an sedimentation with an organic solvent, an ultrafiltration method, a gel filtration, all kinds of chromatography such as a diethylaminoethyl (DEAE)-sepharose chromatography, an anion chromatography or an ion exchange chromatography using lysine such as DIAION HPA-75 (Mitsubishi Chemical Corporation), a cation chromatography using lysine such as S-Sepharose FF (Pharmacia), a hydrophobic chromatography or an affinity chromatography such as butylsepharose or all kinds of electrophoresis such as a SDS-polyacrylamide gel electrophoresis or an electro-focussing electrophoresis.","An affinity chromatography can be accomplished by using antibodies against KIAA1036 polypeptide.","When KIAA1036 polypeptides are produced and accumulated as insoluble polypeptides, cells or fungus bodies are separated as mentioned above and broken into pieces by an appropriate method.","Then a division including the polypeptides is collected.","A collected sample is solubilized with a solubilizer like a surfactant such as sodium lauryl sulfate (SDS) or Sodium N-Dodecanoylsalcosinate (salcosiyl).","After the solubilized solution is diluted or dialyzed to the concentration that a solubilizer is not or almost not included and the polypeptide is constructed to a normal stereo structure, a purification sample can be obtained by a method of separation/purification as mentioned above.","And KIAA1036 polypeptides can be produced as fusion proteins with the other proteins and purified by an affinity chromatography with substances having affinity with the fusion proteins (Yamakawa Akio, “Experimental Medicine”, 13, 469-474 (1995).).","An addition protein used as a fusion protein is, for example, protein A or FLAG (Proceedings of the National Academy of Sciences USA, 86, 8227 (1989)., Genes Development, 4, 1288 (1990)., JP1993-336963 or JP1994-823021).","When protein A are used, fusion proteins with KIAA1036 polypeptides and protein A can be produced and purified by an affinity chromatography with immunoglobulin G. When FLAG peptides are used, fusion proteins with KIAA1036 polypeptides and FLAG can be produced and purified by an affinity chromatography with anti-FLAG antibodies.","KIAA1036 polypeptides can be produced as well as a well-known method with in vitro transcription/translation system (Journal of Biomolecular NMR, 6, 129-134 (1995)., Science, 242, 1162-1164 (1988), or The Journal of Biochemistry, 110, 166-168 (1991).).","KIAA1036 polypeptides can be chemosynthesized on the basis of the amino acid sequence by a chemical synthesis method such as Fmo method (Fluorenylmethyl oxycarbonyl method) or tBoc method (t-butyl oxycarbonyl method), or by a peptide synthetic equipment on the market such as, for example, APEX396 (Advanced Chemtech), 433A (Applied Biosystems), PS3 (Protein Technologies), 9050 (Perseptive) or PSSM-8 (Shimazu corporation).","A structural analysis of KIAA1036 polypeptide can be accomplished by a usual method in the field of protein chemistry, for example, a method described in “A protein structural analysis for gene cloning” (Hisashi Hirano, TOKYO KAGAKU DOZIN Co., LTD., 1993).","The chordin activity of a polypeptide of the present invention can be measured as well as a well-known assay (The Journal of Biological Chemistry, 258, 9893-9898 (1983)., The Journal of Biological Chemistry, 262, 15649-15658 (1987)., The Journal of Biological Chemistry, 273, 58-65 (1998)., The Journal of Biological Chemistry, 273, 433-440 (1998)., The Journal of Biological Chemistry, 273, 12770-12778 (1998)., Archives of Biochemistry and Biophysics, 270, 630-646 (1989)., Archives of Biochemistry and Biophysics, 274, 14-25 (1989), or JP1994-181759).","(3) A Method of Production of a Variant Polypeptide Mutation A deletion, a substitution or an addition of an amino acid of KIAA1036 polypeptide is accomplished by a widely known technique, a site-specific potentially mutagenic method.","A deletion, a substitution or an addition of 1 or few amino acid(s) can be prepared as well as the method described in Molecular Cloning: A Laboratory Manual, Second Edition (1989) (Cold Spring Harbor Laboratory Press.","), Current Protocols in Molecular Biology (1994) (Wiley-InterScience), Nucleic Acids Research, 10, 6487 (1982)., Proceedings of the National Academy of Sciences USA, 79, 6409 (1982)., Gene, 34, 315 (1985)., Nucleic Acids Research, 13, 4431 (1985)., Proceedings of the National Academy of Sciences USA, 82, 488(1985)., Proceedings of the National Academy of Sciences USA, 81, 5662 (1984)., Science, 224, 1431 (1984)., WO85/00817 or Nature, 316, 601 (1985).","(4) A Production of an Antibody Against KIAA1036 Polypeptide (i) A Production of a Polyclonal Antibody Antibodies can be produced by giving all length of KIAA1036 polypeptides, a part of the peptides or polypeptides including a part of the peptides as antigens to a mammal.","The peptide itself and a carrier, for example, a carrier combined with cattle serum albumin (BSA), keyhole limpet hemocyanin (KLH) or bovine thyroglobulin (BTG) can be used as an antigen.","To enhance immune reactions with antigens, for example, a complete Freund adjuvants (CFA) and an incomplete Freund adjuvants (IFA) can be given.","A mouse, a rat, a rabbit, a goat or a hamster can be used as a mammal to immunize.","Polyclonal antibodies can be produced, for example, as well as the method by Lane et al.","(Antibodies: A Laboratory Manual, Second Edition (1989)(Cold Spring Harber Laboratory Press)).","After the first immunization, a mammal is immunized by an appropriate antigen 3 to 10 times at 1- to 2-week intervals.","And then a serum from the mammal is extracted and purified to produce the antibodies.","The antigens are given 3 to 10 times at 1- to 2-week intervals.","A preferable dosage of the antigens is 50 to 100 μg at one time per an animal.","When peptides are used, peptides covalently bonded to appropriate carriers are preferably used as antigens.","Peptides as antigens can be synthesized by a method of genetic engineering or a peptide synthesizer.","Three to seven days after immunization, the blood from venous plexus of eyegrounds is collected and the responsiveness of the serum against the antigens can be measured by an enzyme-liked-immunosorbent assay (Enzyme-liked-immunosorbent assay (ELISA): Igaku-Shoin Ltd. (1976), Antibodies: A Laboratory Manual, Second Edition (1989)(Cold Spring Harbor Laboratory Press).","Blood is collected from a mammal immunized and an antibody titer is measured.","Blood is collected when the immunized animal shows a sufficient antibody titer, and then polyclonal antibodies can be prepared from the serum.","Separation and purification of polyclonal antibodies can be accomplished by an individual or a combination of all kinds of chromatography such as a centrifugal separation, salting-out with ammonium sulfate, precipitation with caplyric acid (Antibodies: A Laboratory Manual, Second Edition (1989)(Cold Spring Harbor Laboratory Press), DEAE-sepharose column, anion exchange column, protein A column or G-column or a gel filter column.","(ii) A Production of a Monoclonal Antibody (a) A Preparation of an Antibody Producing Cell After a sufficient antibody titer in (i) is obtained, spleen or lymph node is extracted from the mammal.","And then a monoclonal antibody producing hybridoma can be obtained by fusing an antibody-producing cell from the spleen or lymph node with a myeloma cell.","As for the myeloma cell, cell strains established from a mouse or a rat can be used.","Cell fusion can be done according to an already known method, for example, a method by Kohler and Milstein (Nature, 256, 495-497 (1975).).","KIAA1036 polypeptides, a part of the peptides or polypeptides including the part of the peptides are immunized to a rat.","Three to seven days after a rat showed a sufficient antibody titer, the rat is immunized with the antigen for the last time, and its spleen is extracted as antibody producing cells.","The spleen is cut to pieces in MEM medium (Nissui Pharmaceutical Co. Ltd.) and untied with tweezers.","And then a precipitant is obtained after a centrifugation at 1,200 rpm for 5 minutes.","Splenocytes is separated by treating the precipitant with Tris-ammonium chloride buffer (pH 7.65) for 1 to 2 minutes to remove red blood cells.","The splenocytes are washed with MEM medium 3 times and are used as antibody producing cells.","(b) A Preparation of a Myeloma Cell An established cell line from mouse or rat is used as a myeloma cell.","For example, it is a myeloma cell of 8-azaguanine resistance mouse (from BALB/c), P3-X63Ag8-U1 strain (described blow as P3-U1) (Current Topics Microbiological Immunology, 81, 1 (1978), or European Journal of Immunology, 6, 511 (1976).","), SP2/0-Ag14 strain (described blow as SP-2) (Nature, 276, 269 (1978).","), P3-X63-Ag8653 strain (described blow as 653) (Journal of Immunology, 123, 1548 (1979).)","or P3-X63-Ag8 (described blow as X63) (Nature, 256, 495 (1975).).","These cell strains are subcultured in a 8-azaguanine medium (a normal medium including 15 μg/ml 8-azaguanine (RPMI1640 medium including 1.5 mM glutamine, 5×10−5M 2-mercaptoethanol, 10 μg/ml gentamysin and 10% FCS made by CSL)) and cultured in a normal medium for 3 to 4 days before cell fusion.","2×107 or more cells are prepared for cell fusion.","(c) A Production of Hybridoma Antibody producing cells prepared in (a) and myeloma cells prepared in (b) are washed with MEM medium or PBS (per 1 L; 1.83 g sodium phosphate dibasic, 0.21 g monobasic potassium phosphate, 7.65 g NaCl, pH 7.2) and mixed as the number of antibody producing cells is 5 to 10 times larger than that of the myeloma cells.","After a centrifugal separation at 1,200 rpm for 5 minutes, a precipitant is obtained.","The precipitated cells are well untied and 0.2 to 1 ml of polyethylene glycol solution (2 g polyethylene glycol-1000 (PEG-1000), 2 ml MEM medium, 0.7 ml dimethyl sulfoxide (DMSO)) per 108 antibody producing cells is added to the cells with stirring at 37° C. And then 1 to 2 ml of MEM medium is added for several times every 1 to 2 minutes.","The solution is prepared with MEM medium to 50 ml in total.","After a centrifugal separation at 900 rpm for 5 minutes, a precipitant is obtained.","100 ml of HAT medium (normal medium including 10−4 M hypoxanthine, 1.5×10−5 M thymidine and 4×10−7 M aminopterin) is added to a precipitant and the precipitant is untied slowly and suspended.","The suspension is poured into the 96-well culture plate at 100 μl per well and cultured at 37° C. in the presence of 5% CO2 for 7 to 14 days.","By the method described in “enzyme immunoassay” (Antibodies: A Laboratory Manual, Second Edition (1989)(Cold Spring Harbor Laboratory Press)), hybridomas producing antibodies specifically reacting with KIAA1036 polypeptides are selected.","(5) an Immunoassay of KIAA 1036 Polypeptide The method to measure polypeptides or a part of the peptides by using antibodies that recognize the polypeptides or a part of the peptides and are directly or indirectly bonded to enzymes, fluorescent substances, radioisotopes or latexes can be used as an immunoassay of KIAA 1036 polypeptides.","The assay is, for example, ELISA or a chemiluminescence method detecting enzyme activities such as horseradish peroxidase or alkaline phosphatase, FITC method detecting fluorescent tags such as luminol or GFP (Green Fluorescence Protein), RIA method detecting radioisotope tags such as 125I or a latex agglutination method detecting binding with latex.","And the assay is, for example, Western blotting or an immune structure dyeing method.","Furthermore, KIAA1036 polypeptides or a part of peptides can be quantitated by the assay.","Antibodies for an immunoassay can be immobilized to a solid phase carrier and the trapped polypeptides can be detected by using secondary antibodies with a reporter group or using reagents.","A competitive method that KIAA1036 polypeptides are labeled with a reporter group, reacted with antibodies and a sample and bonded with immobilized antibodies can be used to detect.","The level of inhibition the binding between the labeled polypeptides and antibodies by KIAA1036 polypeptides of a sample is shown by reactivity with immobilized antibodies of a sample and the concentration of KIAA1036 polypeptides in a sample can be calculated.","Any substance, to which antibodies can attach and which is widely known to a parson having ordinary skill in the art, can be used as a solid phase carrier.","The substance includes, for example, a microtitre plate, a membrane such as a nitrocellulose membrane, bead, disk, glass, glass fiber, plastic material such as latex, polystyrene or polyvinyl chloride.","Magnetic particles or fiber optical sensors (U.S. Pat.","No.","5,359,681) can be used.","A well-known method used in this field can be used as a method which antibodies are immobilized to a carrier.","In this description, “solid phase” means immobilization by a physical method such as adsorption or a chemical binding by a covalent bond between an antibody and a functional group on a carrier.","An antibody and a functional group on a carrier can be bonded directly or through a cross-linking agent.","Immobilization by a physical method can be accomplished by appropriately diluted antibodies contacted with a carrier, preferably, a microtiter plate or a membrane in an appropriate buffer for an appropriate time.","The contact time varies depending on the temperature, but it is typically between about 1 hour and 1 day.","About 10 ng to 1 μg, preferably, about 100 to 200 ng of antibodies is added and immobilized on each well of a microtiter plate made of plastic such as polystyrene or polyvinyl chloride.","Immobilization by a chemical method can be accomplished by a reaction of a carrier and functional groups of antibodies, for example, a reaction of a carrier and a two-functional reagent that reacts with both hydroxyl groups and amino groups and a carrier.","For example, antibodies can be immobilized to a carrier having an appropriate polymer coat with a covalent bond by using benzoquinone or a condensation between aldehyde groups on a carrier and an amine or an active hydrogen on a combination partner.","The method can be accomplished by using for example, Pierce immunotechnology Catalog and Handbook (1991) A12 to A13 as a reference.","A carrier-immobilized antibody is treated to inhibit physical adsorption of other polypeptides by a well-known method for a parson having ordinary skill in the art with an appropriate blocking reagent, for example, cattle serum albumin or Tween 20 (Sigma-Aldrich).","A carrier-immobilized antibody is reacted with a sample and polypeptides of the present invention and antibodies are combined.","A sample can be appropriately diluted with an appropriate diluent, for example, phosphate buffered saline solution (PBS).","A reaction time of a sample and antibodies should be enough to detect the presence of polypeptides of the present invention in a sample obtained from an individual having a cancer, preferably, a time to achieve at least 95% of binding level compared to the level at which bound and not-bound polypeptides are equilibrated.","A time to reach to equilibrium can be easily decided by measuring the binding level by the time.","Substances other than bound polypeptides can be removed by washing a solid carrier with an appropriate buffer, for example, PBS (including 0.1% Tween 20).","Labeled secondary antibodies are reacted with a solid carrier.","The labels are preferably enzymes such as horseradish peroxidase, ground substances, supplemental elements, inhibitors, pigments, radioisotopes, coloring substances or fluorescent substances.","The binding between antibodies and labels can be accomplished by a well-known method for a parson having ordinary skill in the art.","The secondary antibodies are reacted for a sufficient time to bind to complexes, which include immobilized antibodies and polypeptides of the present invention.","An appropriate time can be easily decided by measuring binding level by the time.","The non-binding secondary antibodies can be removed by washing a solid carrier with an appropriate buffer, for example, PBS (including 0.1% Tween 20).","The method of detection of labels of the secondary antibodies is different by a kind of labels.","When radioisotopes are used as labels, detection by a scintillation counter or an autoradiography can be used.","When pigments, coloring substances or fluorescent substances are used as labels, detection by a spectrophotometer can be used.","When enzymes are used as labels, ground substances against the enzymes are added and reacted for a fixed time and the products are detected by a spectrophotometer.","Labels and secondary antibodies can bind directly or indirectly by an avidin-biotin method.","When they bind indirectly, one part of the avidin-biotin is bound to a secondary antibody and another is bound to a label.","KIAA1036 polypeptides can be detected by a flow through test or a strip test.","In a flow through test, a sample is added to a nitrocellulose membrane on which antibodies are immobilized, and when a sample passes through the membrane, polypeptides bind to the immobilized antibodies to form immune complexes.","And when a solution including labeled secondary antibodies pass through the membrane, it binds to the immune complexes.","In a strip test, once a sample is added, the sample pass through a region including labeled antibodies, and polypeptides bind to labeled antibodies to form immune complexes.","When a sample pass through a region including a solid phase antibodies, polypeptides bind to the immune complexes.","The quantity of secondary antibodies detected in the region with immobilized antibodies shows the presence or absence of cancer.","Labels in the detection part can be visualized for confirmation.","If it can not be confirmed, it shows a negative result.","A quantity of antibodies immobilized to a membrane is preferably about 25 ng to 1 μg and more preferably about 50 ng to 1 μg.","(6) An Assay of mRNA Using oligonucleotides prepared from KIAA1036 polynucleotides, mRNAs are quantitated by Northern hybridization or RT-PCR, and then expression level of a gene encoding KIAA1036 polypeptide can be quantitated.","When mRNAs are quantitated with normal or disease model animals, for example, mice, rats, rabbits, sheep, pigs, cattle, cats, dogs or monkeys, an agent such as an anticancer agent is optionally given.","After a fixed time, organs or tissues such as blood, brain, stomach or kidney is isolated and cells are prepared.","mRNAs can be extracted from obtained cells by a widely known and usual method in this field and quantitated/analyzed by RT-PCR or Northern blot hybridization.","When mRNAs from transformants, which express KIAA1036 polypeptides or a part of the peptides, are quantitated, mRNAs can be extracted from the transformants by a widely known and usual method in this field and quantitated/analyzed by RT-PCR or Northern blot hybridization.","(7) Identification of a Structure of the Gene Recently, a lot of sequences of human genome whose functions are unknown are registered on databases.","Therefore, by comparing KIAA1036 polynucleotide sequence and a sequence of human genome registered on databases, human gene encoding KIAA1036 polypeptide can be identified and a structure of the gene can be clarified.","If a genomic sequence, which coincides with cDNA sequence, is registered, a promoter region and a structure of exon and intron of the gene encoding a polypeptide of the present invention can be screened by comparing cDNA sequence and a sequence of the gene.","By using KIAA1036 polynucleotide as a probe and a widely known and usual method in this field (Division of Oncology, Institute of Medical Science, Univ.","of Tokyo, New Protocol for Molecular and Cellular Biology, Shujunsha (1993).","), a promoter region of the gene can be obtained.","Promoter regions are all of the promoter regions concerned with transcription of a gene encoding KIAA1036 polypeptide in mammal cells.","(8) A Method of Detection of Disease KIAA1036 polypeptides can be used for diagnosis of diseases relating to neovaucularization, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer because the expression is increased in angiogenesis models.","Furthermore, because the expression of the polypeptides is increased in ovarian cancer and large bowel cancer tissue, it can be used for diagnosis of these cancers.","Polypeptides of the present invention or polynucleotides such as mRNAs are specifically detected from biological samples, for example, cancer or normal tissue specimen, dissected tissue, blood, lymph node, serum, urine or other tissues obtained from a patient and their homogenates or extracts.","Then presence or absence of cancer in a patient can be decided by comparing the expression level in a patient with a level in a normal person or a normal tissue.","The method of detection of disease includes, for example, an immunoassay or a molecular-biological method of detection of the polypeptides.","An immunoassay of the polypeptides includes, for example, sandwich ELISA.","In this method, a pair of antibodies recognizing different epitopes of the polypeptide are used and one antibody is labeled directly or indirectly with an enzyme.","Another method is a competitive RIA by using the polypeptides labeled with radioisotopes such as 125I and antibodies recognized them.","A molecular-biological method of detection includes, for example, in situ hybridization or PCR.","PCR can be used to amplify cDNA prepared from mRNA isolated from a biological sample from a patient.","A primer for PCR can be chemically synthesized on the basis of DNA sequence of SEQ ID NO: 1.Preferred is Primer1 which is an oligonucleotide sequence of SEQ ID NO: 4,5′-GTTCAGGACTGTCTTTCAGC-3′, or Primer2 which is an oligonucleotide sequence of SEQ ID NO: 5,5′-GTCAATACTGATGGACTTGC-3′.","cDNA amplified by PCR can be separated by gel electrophoresis and then turned visible by the method widely known to a parson having ordinary skill in the art.","Usually, samples are appropriately prepared from a pair of tissues, which are a cancer tissue and a normal tissue from the same individual or different individuals and subjected to PCR.","An amplification reaction by PCR is preferably held with a serial two orders dilution of template cDNAs.","In some dilution points, more than two-fold increase in expression of a cancer sample compared to a normal sample is considered as a positive result.","The presence or absence of cancer is judged by measuring markers of the present invention detected in a normal sample and a patient sample and comparing.","Generally, a mean value of markers from a healthy person without cancer is considered as a cut-off value and the cut-off value is compared to detected value of markers from a patient sample.","Generally, when the detected value is over three times of the standard deviation plus a cut-off value, the sample is considered as a positive about cancer.","And a cut-off value is decided by Receiver Oparator Curve described in Sackett et al., Clinical Epidemiology: A Basic Science for Clinical Medicine 106-107, (1985)(Little Brown and Co).","(9) A Diagnostic Kit KIAA1036 polynucleotides can be used for a diagnosis of neovascularization related disease, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer such as ovarian cancer or large bowel by Northern hybridization or PCR.","And a diagnosis of neovascularization related disease, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer such as ovarian cancer or large bowel can be held by an immunologic method with antibodies against KIAA1036 polypeptides.","Therefore, in the case of a diagnosis with polynucleotides, labeled KIAA1036 polynucleotides are contained in a kit.","And in the case of a diagnosis with antibodies, KIAA1036 polypeptides are contained as standard antigens in addition to antibodies against KIAA1036 polypeptides.","In a kit, a standard curve can be contained.","(10) The Method to Inhibit mRNA Translation Giving KIAA1036 polynucleotides can inhibit the production of KIAA1036 polypeptides.","Namely, by using KIAA1036 polynucleotides, transcription of DNA encoding KIAA1036 polypeptide and translation of mRNA encoding the polypeptide can be inhibited.","(11) The Screening Method of Binding Substances As a reagent for search or screening binding substances to markers of the present invention, KIAA1036 polypeptides are useful.","Namely, the screening method of binding substances to markers of the present invention, which comprises contact between markers and subjects, is provided.","Subjects include, for example, chemical substances synthesized naturally or chemically, proteins, tissue extracts of a mammal such as human, mouse, rat, pig, cattle, sheep or monkey or cell culture.","These subjects are added to markers of the present invention and the mixture is separated with measuring DNA synthesis activity of vascular endothelial cells.","Finally, single ligands can be obtained.","For example, the screening method of binding substances of the present invention includes a method by using KIAA1036 polypeptides or the partial peptides, by constructing an expression system of recombinant polypeptides and using a binding assay with the expression system, by binding to KIAA1036 polypeptides and measuring cell stimulatory activity, for example, activity to promote or inhibit DNA synthesis by bringing BrdU (bromodeoxyuridine) or cell migrating activity or by measuring visually network formation of vascular endothelial cells with microscope.","At first, any polypeptide comprising polypeptide B can be used as polypeptides for the screening method of the binding substances but polypeptides expressed massively with animal cells is preferable.","To produce polypeptide B, the above mentioned expression method is used, but the method is preferable when it includes expressing DNA encoding the polypeptide in mammal cells or insect cells.","Generally cDNA is used as a fragment which encodes a protein to be expressed, but it should not be always cDNAs but fragment from genomic DNA or synthetic DNA can be used.","To introduce DNA fragments encoding KIAA1036 polypeptides into host animal cells and effectively express them, it is preferable that the DNA fragments are inserted downstream of a polyhedrin promoter of nuclear polyhedrosis virus (NPV) of baculovirus which use insects as a host, a promoter from SV40, a promoter of retrovirus, a metallothionein promoter, a human heat-shock promoter, a cytomegalovirus promoter or a SR α promoter.","The examination of quantity and quality of expressed polypeptides can be done by a well-known method.","For example, it is the method described in The Journal of Biological Chemistry, 267, 19555 to 19559 (1992).","Therefore, in a screening method of binding substances of the present invention, polypeptides or the partial peptides purified by the well-known method, cells including the polypeptides or their supernatants can be used as a thing including KIAA1036 polypeptides or the partial peptides.","In a screening method of binding substances of the present invention, when cells including KIAA1036 polypeptides are used, that cells can be immobilized to agarose gel.","The method of immobilization can be held by the well-known method.","Cells including KIAA1036 polypeptides mean host cells with expressing KIAA1036 polypeptides.","For example, Escherichia coli, Bacillus subtilis, yeast, insect cells and animal cells are used as the host cells.","For the screening of binding substances to KIAA1036 polypeptides, appropriate polypeptide fractions and labeled test substances are needed.","As a polypeptide fraction, for example, a natural polypeptide fraction or a recombinant polypeptide fraction having the same activity as a natural one is preferable.","The same activity means, for example, the same ligand binding activity or the same signal transduction activity.","The labeled test substances are, for example, substances labeled with 3H, 125I, 14C or 35S.","For example, for a screening method of binding substances to KIAA1036 polypeptides, at first, a polypeptide sample is prepared by suspending cells including KIAA1036 polypeptides or a membrane fraction of the cells in a buffer which is suitable for a screening method.","Any buffer, which does not inhibit combining a ligand and a polypeptide, for example, a phosphoric acid buffer that is pH 4 to 10, preferably pH 6 to 8 or a Tris-HCl buffer is used as a buffer.","And to decrease non-specific binding, a surfactant, for example, CHAPS, Tween-80 (Kao-Atlas), digitonin or deoxycolate or all kinds of protein, for example, cattle serum albumin or gelatin can be added into the buffer.","Furthermore, to repress degradation of receptors and ligands by protease, a protease inhibitor, for example, PMSF, leupeptin, E-64 (PEPTIDE INSTITUTE, INC.) or pepstatin can be added.","Test substances are labeled with, for example, constant quantity, preferably 5000 cpm to 500000 cpm of 3H, 125I, 14C or 35S are coexistence in 0.01 to 10 ml of the polypeptide solution.","To measure the quantity of non-specific binding (NSB), a reaction tube with excessive amount of unlabeled test substances is also prepared.","The reaction is held at 0° C. to 50° C., preferably at 4° C. to 37° C., for 20 minutes to 24 hours, preferably for 30 minutes to 3 hours.","After the reaction, the solution is filtered by glass fiber filter papers and washed with a suitable quantity of the same buffer.","The radioactivity remaining on the glass fiber filter paper is measured by a liquid scintillation counter or a γ-counter.","Test substances that the count (B-NSB), the result obtained by subtracting quantity of non-specific binding (NSB) from all quantity of binding (B), is over 0 cpm can be selected as binding substances to markers of the present invention.","For the screening of binding substances to markers of the present invention, cell stimulatory activity through the polypeptides, for example, activity to promote or inhibit DNA synthesis by bringing BrdU (bromodeoxyuridine) or cell migrating activity can be measured by well-known method or a measuring kit on the market.","And the activity can be measured by measuring visually network formation of vascular endothelial cells with microscope.","For example, at first, cells including polypeptides are cultured on the multiwell plate.","When the screening of binding substances is held, in advance, medium or buffer changes to fresh medium or appropriate buffer, which is non-toxic to cells and test substances are added.","After incubation for a fixed time, cells are extracted or their supernatants are collected and the products are quantitated by the method for them.","(12) A Kit for Screening of Binding Substances A kit for screening of binding substances bonded to markers of the present invention is a kit comprising cells expressing polypeptide B or the polypeptides or their supernatant fraction including the polypeptides.","The examples of a kit for screening of binding substances are the followings.","(i) A Reagent for Screening of Binding Substances As a screening solution and washing solution, a solution that Hank's Balanced Salt Solution (Invitrogen) added 0.05% cattle serum albumin (SIGMA-Aldrich) is sterilized by filtration with 45 μm filter, and it is kept at 4° C. or prepared just before use.","(a) COS-7 Cells Expressing Polypeptides of the Present Invention are Cultured on a 12-well Plate with 5×105 Cells/well and Cultured at 37° C. for 2 Days.","(b) A Labeled Subject A solution containing commercially available subjects labeled with 3H, 125I, 14C or 35S is kept at 4° C. or 20° C., and diluted to 1 μM with assay buffer just before use.","Subjects with the low solubility to water are dissolved in dimethyl formamide, DMSO or methyl alcohol.","(c) A Non-Labeled Subject The same things as labeled substances are prepared at 100 to 1000 times higher concentration.","(ii) Assay (a) COS-7 cells expressing polypeptides of the present invention cultured on a 12-well plate for tissue culture are washed twice with assay buffer (iml) and then assay buffer is added (490 μl per well).","(b) Labeled subjects (5 μl) are added and the solution is reacted at a room temperature for 1 hour.","Non-labeled subjects (5 μl) are added to measure the quantity of unspecific binding.","(c) The reaction solution is removed and the remaining cells are washed with a washing solution (1 ml) 3 times.","Labeled subjects bound to cells are dissolved with 0.2M NaOH (included 1% SDS) and the solution is mixed with liquid scintillator A (4 ml) (Wako Pure Chemical Industries, Ltd.).","(d) The radioactivity is measured by liquid scintillation counter (Beckman Coulter).","(13) A Screening Method of Binding Activity Regulatory Substances A marker of the present invention is useful as a reagent to search or screen binding activity regulatory substances to the markers.","As a screening method of binding activity regulatory substances of the markers and binding substances, namely, substances changing the binding activity, a recombinant expression system expressing markers of the present invention is constructed and bioassay system or binding assay system is used.","Then substances changing the binding activity between binding substances and markers of the present invention, for example, peptides, proteins, nonpeptide substances, synthetic substances or ferment products can be efficiently screened.","The substances comprise a substance which can enhance or reduce (a) cell stimulatory activity between binding substances and a marker of the present invention, for example, an activity to promote or inhibit DNA synthesis monitored by BrdU (bromodeoxyuridine) incorporation or an activity to promote or inhibit of cell migrating activity, or (b) the binding activity between a binding substance and a marker of the present invention.","Namely, the present invention provides with a screening method of binding activity regulatory substances between binding substances and markers of the present invention characterized by comparing the quantity of binding of binding substances when (i) markers of the present invention and binding substances are contacted and (ii) markers of the present invention, binding substances and subjects are contacted.","A screening method of binding activity regulatory substances of the present invention is characteristic of comparing the quantity of binding between the markers and binding substances by measuring, for example, cell stimulatory activity in the case of (i) and (ii).","A screening method of binding activity regulatory substances of the present invention is, for example, (a) a method to screen binding activity regulatory substances which can change the binding activity between binding substances and KIAA1036 polypeptides, comprising measuring and comparing the quantity of binding of labeled substances to the polypeptides when labeled binding substances are contacted to KIAA1036 polypeptides and when labeled binding substances and subjects are contacted, (b) a method to screen binding activity regulatory substances which can change the binding activity between binding substances and polypeptides of the present invention comprising measuring and comparing the quantity of binding of label binding substances to the polypeptides in the cells or cell culture when cells expressing KIAA1036 polypeptides or the cell culture solution and labeled binding substances are contacted and when labeled binding substances and subjects are contacted or (c) a method to screen binding activity regulatory substances which can change the binding activity between binding substances and KIAA1036 polypeptides comprising measuring and comparing the quantity of binding of label binding substances to the polypeptides when polypeptides secreted in cell culture solutions by culturing transformants having KIAA1036 polynucleotides and label binding substances are contacted and when label binding substances and subjects are contacted.","(14) A Kit for Screening of Binding Activity Regulatory Substances A kit for screening of binding activity regulatory substances changing the binding activity between KIAA1036 polypeptides and binding substances is a kit comprising cells including the polypeptides or a cell culture solution.","The examples of a kit for screening of the present invention are the followings.","(i) A Reagent for Screening of Binding Activity Regulatory Substances As a screening solution and washing solution, a solution that Hank's Balanced Salt Solution (Invitrogen) added 0.05% cattle serum albumin (SIGMA-Aldrich) is sterilized by filtration with 45 μm filter, and it is kept at 4° C. or prepared just before use.","(a) KIAA1036 Polypeptide COS-7 cells expressing KIAA1036 polypeptides are cultured on a 12 well plate with 5×105 cells/well and cultured at 37° C. for 2 days.","(b) A Labeled Subject A solution containing commercially available subjects labeled with 3H, 125I, 14C or 35S is kept at 4° C. or 20° C., and diluted to 1 μM with assay buffer and prepared just before use.","Subjects with the low solubility to water are dissolved in dimethyl formamide, DMSO or methyl alcohol.","(c) A Non-Labeled Subject The same things with labeled substances are prepared at 100 to 1000 times higher concentration.","(ii) An Assay (a) COS-7 cells expressing polypeptides of the present invention cultured on a 12-well plate for tissue culture are washed twice with assay buffer (1 ml) and then assay buffer is added (490 μl per well).","(b) Labeled subjects (5 μl) are added and the solution is reacted at room temperature for 1 hour.","Non-labeled subjects (5 μl) are added to measure the quantity of non-specific binding.","(c) The reaction solution is removed and the remaining cells are washed with a washing solution (1 ml) 3 times.","Labeled subjects binding to cells are dissolved with 0.2M NaOH (included 1% SDS) and the solution is mixed with liquid scintillator A (4 ml) (Wako Pure Chemical Industries, Ltd.).","(d) The radioactivity is measured by liquid scintillation counter (Beckman Coulter).","(15) A Screening Method of Expression Regulatory Substances Polynucleotide A, polypeptide B or antibodies against the polypeptides are useful as a reagent to search or screen expression regulatory substances to markers of the present invention.","Namely, a screening method of expression regulatory substances of the present invention is, for example, a screening method of expression regulatory substances of markers of the present invention by measuring quantity of mRNAs or proteins of KIAA1036 polypeptides or the partial peptides included in (i) blood of a non-human mammal, a specific organ, a tissue or cells isolated from the organ or (ii) transformants and so on.","(16) A Kit for Screening of Expression Regulatory Substances A kit for screening of expression regulatory substances of KIAA1036 polypeptides or its salt is a kit comprising cells expressing polypeptide A or polypeptide B or cell culture solutions including the polypeptides.","The Example of a kit for screening of the present invention is the following.","(17) A Kit for Screening on the Basis of the Immunologic Assay An assay of KIAA1036 polypeptides is, for example, sandwich ELISA using two kinds of monoclonal antibodies that their epitopes are different in antibodies reacting with the polypeptide in the liquid phase, or a radioimmunoassay using KIAA1036 polypeptides labeled with a radioisotope such as 126I and antibodies specifically recognizing it.","Therefore, in a case of diagnosis with antibodies, KIAA1036 polypeptide is included as standard antigens in addition to antibodies of the present invention.","Furthermore, standard curve can be included in a kit.","(18) A Kit for Screening on the Basis of a Molecular-Biological Assay The expression level of DNA encoding KIAA1036 polypeptide can be quantitated in mRNA level by Northern hybridization or PCR using oligonucleotides prepared from KIAA1036 polynucleotides.","For example, (i) an agent, for example, an anticancer agent is given into a normal or a disease model non-human mammal (e.g., mouse, rat, rabbit, sheep, pig, cattle, cat, dog or monkey) and after a fixed time, blood, a specific organ, for example, brain, kidney, colon, ovary, tissues or cells isolated from an organ are obtained.","mRNAs of a polypeptide of the present invention or the partial peptide included in the obtained cells can be for example, extracted by the extraction method widely known in this field, quantitated by the method such as TaqManPCR or analyzed by Northern blot widely known, or (ii) transformants expressing KIAA1036 polypeptides or the partial peptides are produced by the above mentioned method and mRNAs of KIAA1036 polypeptides or the partial peptides included in the transformant can be quantitated/analyzed.","And in a case of diagnosis with polynucleotides, labeled KIAA1036 polynucleotides are included in a kit.","(19) A Pharmaceutical Composition A pharmaceutical composition comprising at least one selected from substances such as polynucleotide A, a complementary polynucleotide to the polynucleotide, polypeptide B, its antibody or a gene therapeutic vector having polynucleotide A can be given independently as a therapeutic agent.","But preferred is usually that a pharmaceutical composition is mixed with one or more pharmaceutically acceptable carrier(s) and provided as a pharmaceutical preparation produced by the any method widely known in the technical field of galenical pharmacy.","Because KIAA1036 inhibit DNA synthesis in vascular endothelial cells and cell migrating activity, DNA synthesis inhibitors and cell migrating activity inhibitors are included.","And substances obtained by a screening method or a kit for screening of the present invention and their salts are included.","The substance is, for example, (a) a substance enhancing or reducing cell stimulation activity through binding between binding substances and KIAA1036 polypeptides, for example, DNA synthesis activity, migrating activity, network formation activity of vascular endothelial cells, (b) a substance enhancing or reducing a binding activity between a binding substance and KIAA1036 polypeptide or (c) a substance enhancing or reducing an expression level of KIAA1036 polypeptide.","The substance is, for example, a low-molecular weight compound, a peptide, a protein, a nonpeptide substance, a synthetic substance or a ferment product.","These substances can be new or well-known substances and natural or synthetic substances.","Because agonists against KIAA1036 polypeptides have the same activity as bioactivation of binding substances against KIAA1036 polypeptides, they are useful as safe and low-toxic drugs for the binding substance activity.","Because antagonists against KIAA1036 polypeptides can inhibit bioactivation of binding substances against KIAA1036 polypeptides, they are useful as safe and low-toxic drugs, which inhibit the binding substance activity.","Substances enhancing binding activity between binding substances and KIAA1036 polypeptides are useful as safe and low-toxic drugs to enhance bioactivation of binding substances against KIAA1036 polypeptides.","Substances reducing binding activity between binding substances and KIAA1036 polypeptides are useful as safe and low-toxic drugs to reduce bioactivation of binding substances against KIAA1036 polypeptides.","And gene therapeutic expression vectors expressing KIAA1036 are included.","Because KIAA1036 can be a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, it is suggested that KIAA1036 relates to those diseases and they are useful for gene therapies of these diseases.","The gene therapeutic expression vectors are expression vectors having a part or all of a polynucleotide of KIAA1036 and when they are introduced into cells or tissues, they can normalize disorders which cause some natural or acquired disease at a gene level.","In other words, the vectors can compensate for cells with normal genes, repair/modify defects of genes and regulate expression of genes.","(20) A Therapeutic Agent A pharmaceutical composition comprising polypeptide B, an antibody against the polypeptide, polynucleotide A, complementary polynucleotide to the polynucleotide, a binding substance, a bond regulatory substance, an expression regulatory substance, a DNA synthesis inhibitor, a cell migrating activity inhibitor or a gene therapeutic vector can be given independently as a therapeutic agent.","But preferred is usually that a pharmaceutical composition is mixed with one or more pharmaceutically acceptable carrier(s) and provided as a pharmaceutical preparation produced by the any method widely known in the technique field of galenical pharmacy.","The most effective route of administration in treats is desirable as a route of administration, for example, an oral administration or a non-oral administration such as intraoral, tracheobronchial, intrarectal, subcutaneous, intramuscular or intravenous.","A form of administration is, for example, spray, capsule, tablet, granule, syrup, emulsion, suppository, injection, ointment or tapes.","An appropriate pharmaceutical preparation for an oral administration is, for example, emulsion, syrup, capsule, tablet, powder or granule.","For example, a liquid preparation such as emulsion or syrup can be produced by using water, saccharide such as sucrose, sorbitol or fructose, glycol such as polyethylene glycol or propylene glycol, oil such as sesame oil, olive oil or soybean oil, antiseptic such as p-hydroxy ester benzoate or flavor such as strawberry or peppermint as a excipient.","Capsule, tablet, powder or granule can be produced by using a vehicle such as lactose, dextrose, sucrose or mannitol, a disintegrating agent such as starch or sodium alginate, a lubricant such as magnesium stearate or talc, a binding agent such as polyvinyl alcohol, hydroxypropylcellulose or gelatin, a surface active agent such as fatty acid ester or a plasticizer such as glycerol as a excipient.","An appropriate pharmaceutical preparation for non-oral administration is, for example, injection, suppository or spray.","For example, an injection is prepared by using a carrier comprising salt solution, dextrose solution or their mixture.","A suppository is prepared by using a carrier such as cacao butter, fat hydride or carboxylic acid.","And a spray is prepared by using the substance or a carrier, which does not stimulate oral cavity and airway mucous membrane of a recipient and spread the substance as microparticles to become easy to absorb.","The carrier includes, for example, lactose or glycerol.","A pharmaceutical preparation such as aerosol or dry powder is possible if a nature of the substance and that of the used carrier are appropriate for it.","And in these non-oral agents, a component used as an excipient in oral agents can be added.","A substance obtained by using a screening method or a kit for screening of the present invention or the salt is a binding substance, a binding activity regulatory substance, an expression regulatory substance, a DNA synthesis inhibitor or a cell migrating activity inhibitor, for example, (a) a substance enhancing or reducing cell stimulation activity through bonds between binding substances and polypeptide B, for example, an activity to promote or inhibit DNA synthesis, which can be monitored by BrdU (bromodeoxyuridine) uptake, or an activity to promote or inhibit of cell migrating activity, (b) a substance enhancing or reducing a binding activity between binding substances and polypeptide B or (c) a substance enhancing or reducing an expression level of a marker of the present invention.","The substance is a low-molecular weight compound, peptide, protein, non-peptide substance, a synthetic substance or a ferment product.","These substances can be new or well-known substances and natural or synthetic substances.","Because agonists against KIAA1036 polypeptides have the same activity as bioactivation of binding substances against KIAA1036 polypeptides, they are useful as safe and low-toxic drugs for the binding substance activity.","Because antagonists against KIAA1036 polypeptides can inhibit bioactivation of binding substances against KIAA1036 polypeptide or something like this, they are useful as safe and low-toxic drugs, which inhibit the binding substance activity.","Substances enhancing binding activity between binding substances and KIAA1036 polypeptides are useful as safe and low-toxic drugs to enhance bioactivation of binding substances against KIAA1036 polypeptides.","Substances reducing binding activity between binding substances and KIAA1036 polypeptides are useful as safe and low-toxic drugs to reduce bioactivation of binding substances against KIAA1036 polypeptides.","Substances reducing binding activity between binding substances and KIAA1036 polypeptides are useful as safe and low-toxic drugs to reduce bioactivation of binding substances against KIAA1036 polypeptides.","And therapeutic agent KIAA1036 polypeptides for any kinds of diseases comprising substances changing the expression level of KIAA1036 polypeptides seem to have any important role, for example, a central nervous system as described above.","Therefore an expression regulatory substance changing the expression level of KIAA1036 polypeptides or the partial peptide can be used as a therapeutic agent for a disease related insufficiency of KIAA1036 polypeptides.","When the substance is used as a therapeutic agent for a disease relating insufficiency of KIAA1036 polypeptide, it can be turned a pharmaceutical preparation by the stereotyped method.","For example, the substance can be used orally as a tablet with sugar-coating, a capsule, an elixir or a microcapsule as occasion demandsor parentally as a sterile solution with water or the other pharmaceutically acceptable solution or an injection of suspension.","For example, the substances can be produced by mixing as the unitary dose form required to accomplish popularly acceptable pharmaceutical preparation with physiologically acceptable well-known carrier, spice, vehicle, antiseptic, stabilizer or binding agent.","The amount of active principle in these pharmaceutical preparations is decided to be able to obtain an appropriate capacity in a directed range.","An excipient, which can mix with tablet or capsule, is, for example, a binding agent such as gelatin, corn starch, tragacanth or Arabian gum, a vehicle such as crystal cellulose, a swelling agent such as corn starch, gelatin or alginic acid, a lubricant agent such as magnesium stearate, a sweetening agent such as sucrose, lactose or saccharin or a spice such as peppermint or cherry.","In the case where the dosage form is a capsule, the above type of ingredients and a liquid carrier such as fat and oil can be included.","An aseptic composition for the injection can be prescribed by the usual pharmaceutical preparation, for example, by dissolving or suspending an activity substance in vehicle such as water for the injection or natural plant oil such as sesame oil or coconut oil.","An aqueous solution for the injection is, for example, an isotonic solution such as D-sorbitol, D-mannitol or sodium chloride including physiological salt solution, dextrose or adjuvant.","It can be used with appropriate solubilizing agent, for example, alcohol such as ethanol, propylene glycol or polyethylene glycol, a nonionic surface-active agent such as polysorbate 80 or HCO-50.Oil solution is, for example, sesame oil or soybean oil and it can be used with a solubilizing agent such as benzyl benzoate or benzyl alcohol.","And the above preventive/therapeutic agent can be mixed with, for example, a buffer such as phosphate buffer or sodium acetate buffer, a soothing agent such as benzalkonium chloride or procaine hydrochloride, a stabilizer such as human serum albumin or polyethylene glycol, a preservatives such as benzyl alcohol or phenol or an antioxidant.","A prepared injection solution is usually filled up into an appropriate ampoule.","(21) A Method of Treating Because the above-mentioned pharmaceutical composition can be a therapeutic agent for vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, a method of treating disease such as vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer is provided by giving that pharmaceutical composition by an appropriate method.","It is desirable that the above-mentioned pharmaceutical composition is used in the most effectively medication method in treats.","And the method is, for example, an oral administration or a non-oral administration such as intraoral, tracheobronchial, intrarectal, subcutaneous, intramuscular or intravenous.","A therapeutic agent as described above is safe and low-toxic and can be given to, for example, a mammal such as human, rat, mouse, rabbit, sheep, pig, cattle, cat, dog or monkey.","A dosage of a preventive or therapeutic agent per 1 time varies depending on medication subject, an object organ, a condition of a disease or a medication method.","But in the case of a oral administration, for example, for a patient with hypertonia whose body weight is 60 kg, usually approximately 0.1 to 100 mg per a day, preferably approximately 1.0 to 50 mg, more preferably approximately 1.0 to 20 mg are given.","For example, in the case of an injection of parentally giving, for example, for a patient with hypertonia whose body weight is 60 kg, usually approximately 0.01 to 30 mg per a day, preferably approximately 0.1 to 20 mg, more preferably approximately 0.1 to 10 mg are given by an intravenous injection.","In the case of the other animal, the amount converted into the amount per kg of body weight can be given.","The dosage or times of a dosage varies depending on treating effect targeted, medication method, treating period, age or body weight, but usually it is 10 μg to 8 mg/kg per a day for an adult.","EXAMPLES The following examples are provided to exemplify and do not restrict the present invention.","As a genetic engineering method, if there is not a special provision, the method described in Molecular Cloning: A Laboratory Manual, 2nd Edition (Cold Spring Harbor Laboratory), is used.","Example 1 An Identification of Gene Whose Expression in Human Umbilical Vein Endothelial Cells (HUVEC) is Enhanced by the Stimulation of Vascular Endothelial Cell Propagation Factors (VEGF) Three Endocell HUVEC (KURABO) which is a human umbilical vein vascular endothelial cell were thawed and then inoculated on 16 culture plates coating with collagen (IWAKI) poured EBM medium including 10% fetal calf serum (FCS) (Sanko Junyaku).","After culturing at 37° C. for 3 days, the medium was changed with the same medium and cultured for 2 days.","After the cells reached to confluence, the medium was changed with M199 medium including 1% FCS (Nissui) and cells were cultured for 24 hours.","For a culture with VEGF treatment, the medium was changed to M199 medium including 1% FCS and 1 nM human VEGF165 (R&D Systems).","For a culture without VEGF (a control), medium was changed with the same medium not including human VEGF165.Cells were cultured at 37° C. and collected after 0 hour, 0.5 hours, 2 hours, 6 hours, 12 hours and 24 hours.","Collected cells were washed with PBS twice and total RNA were prepared by ISOGEN (Nippon Gene) and mRNAs were prepared by oligo dT cellulose column.","These mRNAs were analyzed by LifeArrayhumanUniGENE (Incyte Genomics) through Gene Expression Microarray analysis on assignment of Incyte Genomics.","As a result, for 6393 human genes, gene expression level was calculated as a ratio of a expression level in VEGF-treated cells compared to one in untreated cells in each time point.","Among these genes, the expression level of a gene GenBank ID: AF055021, whose function was unknown, was increased by treatment with VEGF reaching the expression level of was 3.8 fold after 24 hours.","Because a protein translated region was unclear from the information of sequence of AF055021 gene in a database of NCBI (National Center for Biotechnology Information), a base sequence of the same gene was searched with AssEst (maze) database and a consensus sequence of EST cluster was obtained.","Based on this consensus sequence, homology search using Blastx database in AssEst revealed that it encoded KIAA1036 polypeptide (SEQ ID NO: 2) whose function was unknown.","Additionally, it had 58% homology with a polypeptide of AK022567 whose function was unknown (FIG.","1).","KIAA1036 gene and AK022567 gene on NCBI database have open reading frames corresponding to 365 and 290 amino acid residues respectively, which are initiated from translation initiation codons and they are seem to be almost full length cDNA.","A polypeptide of AK022567 was already published internationally as a secretory protein (WO99/3881).","And KIAA1036 is predicted as a secretory protein because the homology of their amino acid sequences is 58%.","Example 2 Expression of KIAA1036 in Human Normal Tissues Cloning of KIAA1036 gene from human umbilical vein vascular endothelial cell was tried.","Two kinds of oligonucleotides were synthesized as primers for PCR on the basis of a nucleic acid sequence of 3′ untranslated region of KIAA1036 gene from NCBI database.","Primer1: 5′-GTTCAGGACTGTCTTTCAGC-3′ (SEQ ID NO: 4) Primer2: 5′-GTCAATACTGATGGACTTGC-3′ (SEQ ID NO: 5) Total RNA was prepared according to an attached manual with RNeasy Midi (QIAGEN) from HUVEC which are human umbilical vein vascular endothelial cells.","Single strand cDNA fragments were prepared from the obtained total RNA (10 μg) with SuperscriptII (Invitrogen).","The obtained single strand cDNAs were used as templates and gene fragments were amplified with Primer1 (SEQ ID NO: 4) and Primer2 (SEQ ID NO: 5) by PCR.","To one fiftieth of single strand cDNAs, was added TaKaRa Ex Taq (TaKaRa) (1.25 unit), Primer1 and Primer2 (1 μM each), a buffer (Mg2+ plus) for PCR reaction attached TaKaRa Ex Taq (5 μl) and dNTP Mixture whose final concentration was 200 μM.","Distilled water was added to the mixture to be 50 μl in total.","This was how samples for PCR were prepared.","PCR reaction was held under the condition that is 94° C. for 3 minutes was once and the cycle (94° C. for 1 minute, 50° C. for 1 minute and 72° C. for 1 minute) was 30 times and then 72° C. for 5 minutes was once.","The amplified cDNA fragment by the PCR reaction was separated by an agarose electrophoresis and 750 bp cDNA fragment (SEQ ID NO: 6) was collected.","The obtained cDNA fragment was digested with restriction enzymes, ApaI, HincII and PstI and the lengths of cDNA fragments separated by an agarose gel electrophoresis were measured.","And the collected cDNA fragment was confirmed as a target.","The cDNA fragment (50 ng) amplified with Primer1 and Primer2 was labeled with 32P using [α-32P] dCTP (3000 Ci/m mol, 10 mCi/ml) (Amersham Pharmacia Bioteque) by an attached manual with Rediprime II DNA Labeling System (Amersham Pharmacia Bioteque).","Northern blot was held with these cDNA fragments labeled with 32P as a probe.","Nylon membrane of Multiple Tissue Northern (MTN™) Blots Human 12-Lane MTN Blot (Clontech) was immersed with GMC buffer (including 0.5M phosphoric acid buffer, pH 7.2, 1% cattle serum albumin, 1 mM EDTA and 7% SDS) and prehybridization was held at 65° C. for 1 hour.","After that, the nylon membrane was immersed with GMC buffer containing probes labeled with 32P and hybridization was held at 65° C. for 16 hours.","After hybridization, the nylon membrane was washed with 2×SSC (1×SSC was 15 mM sodium citrate, 150 mM NaCl, pH 7.0) at a room temperature for 5 minutes, 1×SSC (including 0.2% SDS) at 50° C. for 30 minutes and 0.5×SSC (including 0.2% SDS) at 50° C. for 30 minutes one after another and dried with air.","Subsequently, this nylon membrane was put on an imaging plate cassette (FUJI FHOTO FILM CO., LTD.) with an imaging plate (FUJI FHOTO FILM CO., LTD.).","Image analysis was held with image analyzer FLA3000 (FUJI FHOTO FILM CO., LTD.).","As a result, signals which showed about 6 kb length were detected from brain, heart, skeletal muscle, spleen, kidney, liver, small intestine and placenta and the strong expression was selectively observed in brain and placenta.","Example 3 Expression of KIAA1036 in Human Umbilical Vein Endothelial Cells As the VEGF-induced expression of KIAA1036 was originally found by Gene Expression Microarray Analysis using DNA chip as described in Example 1, the result was confirmed by Northern blot with KIAA1036 probe and total RNA prepared from VEGF-treated HUVEC.","Endocell HUVEC (KURABO) which are human umbilical vein endothelial cells were cultured in M199 medium containing 1 nM human VEGF165 (R&D Systems) and 1% FCS as described in Example 1.After 0, 0.5, 1, 2, 6, 12 and 24 hours of culture, total RNA was prepared from these cells according to an attached manual with RNeasy Midi (QIAGEN).","After electrophoresis of each RNA sample (5 μg), the samples were transferred to a nylon membrane, Hybond-XL (Amersham Pharmacia Bioteque).","That nylon membrane was immersed with GMC buffer and prehybridization was held at 65° C. for 1 hour.","After prehybridization, the nylon membrane was immersed with GMC buffer containing probes labeled with 32P as Example 2 and hybridization was held at 65° C. for 16 hours.","After hybridization, the nylon membrane was washed with 2×SSC at a room temperature for 5 minutes, 1×SSC (including 0.2% SDS) at 50° C. for 30 minutes and 0.5×SSC (including 0.2% SDS) at 50° C. for 30 minutes one after another and dried with air.","Subsequently, this nylon membrane was put on an imaging plate cassette (FUJI FHOTO FILM CO., LTD.) with an imaging plate (FUJI FHOTO FILM CO., LTD.).","Image analysis was held with image analyzer FLA3000 (FUJI FHOTO FILM CO., LTD.).","Next, probes were dehybridized by immersing the same nylon membrane in a bath with boiling water and Northern blot was held with human β-actin cDNA fragments labeled with 32P as Example 2 as probes.","Signal intensities of KIAA1036 gene were normalized with that of β-actin gene at each time point.","And then a ratio of the expression level of KIAA1036 in each time point to 0 hour was calculated.","As a result, the expression was inhibited at 0.5 to 2 hours after adding VEGF but a high gene expression, which was 3.2 fold, was induced after 24 hours.","This result was same as that obtained by DNA chip (FIG.","2).","Example 4 Expression of KIAA1036 in Human Cancer Tissues The difference of expression levels of the genes in a normal tissue and a cancer tissue of all kinds of organ from human was analyzed.","Matched Tumor/Normal Expression Array (Clontech), which is a nylon membrane with cDNAs synthesized from a cancer tissue and a normal tissue of chest, uterus, colon, stomach, ovary, lung, kidney, rectum, cervix, small intestine or prostate of the same patient, was immersed with GMC buffer containing probes labeled with 32P as Example 2 and hybridization was held at 65° C. for 16 hours.","After hybridization, the nylon membrane was washed with 2×SSC at a room temperature for 5 minutes, 1×SSC (including 0.2% SDS) at 50° C. for 30 minutes and 0.5×SSC (including 0.2% SDS) at 50° C. for 30 minutes one after another and dried with air.","Subsequently, this nylon membrane was put on an imaging plate cassette (FUJI FHOTO FILM CO., LTD.) with an imaging plate (FUJI FHOTO FILM CO., LTD.).","Image analysis was held with image analyzer FLA3000 (FUJI FHOTO FILM CO., LTD.).","After hybridization of the nylon membrane, signal intensities were measured with β-actin genes labeled with 32P as probes as described in Example 3.Signal intensities of KIAA1036 was normalize with signal intensities by β-actin probe in each organ, and a ratio of cancer tissues to normal tissues in the same patient was calculated.","Then a change of an expression level of the genes in all kinds of cancer tissue was analyzed.","As a result, expression of KIAA1036 genes were increased in cancer tissues of all 4 patients with ovarian cancer and 11 patients with colon cancer patient (FIG.","3).","Example 5 Expression of KIAA1036 in COS Cells cDNA encoding the polypeptide that 3×FLAG sequence was added to a carboxy-terminal of KIAA1036 amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 was inserted to a expression vector in animal cells and expressed in COS cells.","A base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 encoding KIAA1036 was inserted in the NotI/XbaI sites of an expression vector in animal cells, p3×FLAG-CMV-13 and p3×FLAG-CMV-14(SIGMA-Aldrich) and plasmids, pFLAG13-1036 and pFLAG14-1036 which express polypeptides that 3×FLAG was added to carboxy- or amino-terminals of KIAA1036, respectively, were constructed.","COS-7 cells (African green monkey kidney cells (SV40-transformed cells)) were used as host of pFLAG13-1036 and pFLAG14-1036 and transformation was operated by an attached manual with FuGENE6 reagent (Roche Diagnostics).","COS-7 cells (5×105 cell/well in 60 mm dishes) were cultured in D-MEM medium containing 10% FCS at 37° C. for 16 hours.","After medium was changed, pFLAG13-1036 (2 μg) and FuGENE6 (10 μl) were added to the cells and the cells were cultured for 48 hours.","pFLAG14-1036 and p3×FLAG-CMV-14 without KIAA1036 cDNA as a control were also used in a same method to make transfectants and the transfectants were cultured.","Expression of KIAA1036 polypeptide fused with FLAG was analyzed by Western blotting, a method of Laemli.","The culture supernatants (10 μl) of all kinds of COS-7 transfectants were subjected to SDS-PAGE and electrically transfered to a nitrocellulose membrane in a nitrocellulose membrane kit (TEFCO).","The nitrocellulose membrane was immersed with a blocking reagent (TBS, pH 7.6; including 5%(w/v) skim milk) and shook at a room temperature for 1 hour.","And the membrane was immersed in TBS reagent (pH7.6; including 5%(w/v) skim milk) containing anti-FLAG M2 antibody labeled with peroxidase (HRP) (SIGMA-Aldrich), put static at a room temperature for 1 hour.","Then the nitrocellulose membrane was washed with a washing reagent (TBS, pH7.6; including 0.05%(v/v) TritonX-100) for 10 minutes 3 times.","The nitrocellulose membrane and ECL Plus Western Blotting Detection Reagents (Amersham Pharmacia Bioteque) were reacted at a room temperature.","The nitrocellulose membrane and X-ray film X-OMAT AR (Kodak) were put on X-OMAT cassette (Kodak) immediately and exposed for a few minutes.","As a result, expected bands, which show about 50 kDa, from the culture supernatant of COS cells transfected with pFLAG13-1036 and pFLAG14-1036 were detected but the band was not detected in a control.","Therefore, it was confirmed that KIAA1036 gene encodes a KIAA1036 protein, which is able to express, and that KIAA1036 was a secretory protein in animal cells.","Example 6 Expression of KIAA1036 Protein in a Baculovirus System Expressions in a baculovirus system were operated with Bac-to-Bac Baculovirus Expression System (Invitrogen) according to an attached manual.","cDNA fragments encoding KIAA1036 with 3×FLAG at the carboxy-terminal described in Example 5 was inserted in NotI/XhoI sites of a plasmid pFastBac1, an expression plasmid for insect cells, to make pFASTBac1036.The recombinant plasmid was introduced in Max Efficiency DH10 Bac Competent Cells and cDNA was transferred into baculovirus genome (bacmid DNA) to give rise to a recombinant bacmid DNA.","The recombinant bacmid DNA was introduced in Sf9 (ATCC: CRL-1711, ovary cell) cell with CellFECTIN Reagent (Invitrogen) to obtain a recombinant baculovirus.","A culture supernatant (0.5 ml) of Sf9 transfectants was added into a culture medium (50 ml) of Sf9 cells with vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or malignant neoplasma (The concentration was 1.5×106 cells/ml) and cultured at 28° C. for 96 hours.","The transformed cells were collected by centrifugation.","Expression of the FLAG-tagged KIAA1036 was confirmed by Western blotting as described in Example 5.The collected transformants were washed once with PBS (5 ml, pH7.4).","After washing, the cells were suspended in Lysis reagent (5 ml) (including 50 mM Tris-HCl, pH7.4; 0.15M NaCl, 0.1 mM EDTA/2Na, 0.1 mM EGTA, 1 mM DTT, 0.1 mM Amidinophenyl methansulfonyl fluride hydrochloride or 0.1% NP-40), treated with ultrasonic waves on ice for 15 seconds for 4 times by MICROCON (HEART SYSTEMS) and separated by centrifugation at 14,500×g for 20 minutes.","The same amount of Lysis reagent was added to the supernatant to make it a crude protein solution.","After a crude protein solution (10 μl) was subjected to SDS-PAGE, it was electrically transferred on a nitrocellulose membrane in nitrocellulose membrane kit (TEFCO).","The nitrocellulose membrane was immersed in a blocking reagent (TBS, pH7.6; including 5%(w/v) skim milk) and shook at a room temperature for 1 hour.","After that, it was immersed in a TBS reagent (pH7.6; including 5%(w/v) skim milk) containing anti-FLAG M2 antibody labeled with peroxidase (HRP) (SIGMA-Aldrich), put static at a room temperature for 1 hour.","And then the nitrocellulose membrane was washed with the washing reagent (TBS, pH7.4; including 0.05%(v/v) TritonX-100) for 10 minutes 3 times.","The nylon membrane and ECL Plus Western Blotting Detection Reagents (Amersham Pharmacia Bioteque) were reacted at a room temperature.","The nitrocellulose membrane and X-ray hyper film ECL (Amersham Pharmacia Bioteque) were put on X-OMAT cassette Kodak) immediately and exposed for a few minutes.","KIAA1036 FLAG proteins were purified by an affinity chromatography with anti-FLAG antibodies.","After an ANTI-FLAG M2-Agarose (SIGMA-Aldrich) affinity column was equilibrated with TBS, pH7.4 (including 0.1% NP-40), a crude protein solution was added and FLAG proteins were separated.","The columns were then washed with TBS (pH7.4; including 0.1% NP-40) and the protein was eluted with Gly-HCl (pH3.5; including 0.1% NP-40) to be 1 ml/fraction.","Each fraction was neutralized with 1M Tris-HCl (pH8.0; 20 μl) to make it an affinity purification fraction.","A crude protein fraction (10 μl) and an affinity purified fraction (10 μl) were subjected to SDS-PAGE and the gel was stained by Bio-Safe Coomassie (BIO-RAD).","As a result, an expected band with about 50 kDa was detected.","Furthermore, after SDS-PAGE, the purified fraction was electrically transferred to PVDF membrane (BIO-RAD) and a band stained with Bio-Safe Coomassie (BIO-RAD) was cut off, and then a sequence of an amino terminal was analyzed by an amino acid sequencer, Procise Model 491 (Applied Biosystems).","As a result, an amino acid sequence as set forth in Pro of 2-Gly of 10 of SEQ ID NO: 2 was detected and it was confirmed that the expressed protein was KIAA1036.Example 7 A Blocking Effect of DNA Synthesis by KIAA1036 in Vascular Endothelial Cells To examine an effect of KIAA1036 protein to the proliferation of vascular endothelial cells, BrdU (bromodeoxyuridine) incorporation was measured in HUVEC cells treated with VEGF in the presence (a treated group) or the absence (a control group) of KIAA1036.A KIAA1036 protein, which was expressed by a baculovirus expression system in Example 6 and purified by an affinity chromatography, was used.","Incorporation of BrdU into cells was measured with cell propagation ELISA and a BrdU chemiluminescence kit according to an attached manual.","Endocell HUVEC (KURABO) which are human vascular endothelial cells (3000 cells/plate) were cultured on collagen-coated plates (IWAKI) with M199 medium (Nissui) containing 5% fetal calf serum (FCS) at 37° C. for 24 hours.","For the VEGF-treated group, medium was changed to M199 medium including 1% FCS, 1 nM human VEGF165 (R&D Systems) and 0, 1, 10 nM KIAA1036 protein.","For the VEGF-untreated group (control group) medium was changed to the same mediums without human VEGF165 containing 0, 1 or 10 nM KIAA1036 protein.","A solution labeled with BrdU (0.02 ml/well) was added and the cells were cultured at 37° C. for 12 hours.","The cultured cells were put static at a room temperature for 1 hour to immobilize and an anti-BrdU antibody solution labeled with POD (0.1 ml/well) was added and the cells were put static at a room temperature for 1 hour.","The cells were washed with a washing buffer for 5 minutes 3 times and a luminecsence substrate (0.1 ml/well) was added.","After 3 minutes of shaking, the instantaneous fluorescence for 5 seconds was measured by Luminescencer JNR (ATTO) within 10 minutes.","As a result, VEGF promoted DNA synthesis of HUVEC, but KIAA1036 protein doses-dependently inhibited it (FIG.","4).","Example 8 A Blocking Effect to the Migration of Vascular Endothelial Cells by KIAA1036 Protein An effect to the migration of vascular endothelial cells of KIAA1036 protein was examined by a modified Boyden chamber method.","To the lower chambers of Transwell (Corning coaster), M199 medium (600 μl /well) containing 0.25 nM human VEGF165 (R&D Systems) and 1 or 10 nM KIAA1036 protein was added for samples treated with VEGF (test group), and M199 medium (600 μl/well) containing 1 or 10 nM KIAA1036 protein without VEGF was added for a control group.","Endocell HUVEC (KURABO) (4×104 cells/well) were cultured in M199 medium containing 5% FCS for 16 hours and then seeded in the upper chambers and cultured at 37° C. for 4 hours.","Filters of Transwell were immobilized with Diffquick fix reagent (Sysmex) and then stained with Diffquick stain (Sysmex).","After wiping the side of the upper chamber of filter, the cell migration ability toward VEGF was measured by counting the number of the cells migrating to the lower part.","As a result, VEGF promoted migration of HUVEC, but KIAA1036 protein significantly inhibited it (FIG.","5).","Example 9 A Blocking Effect to Network Formation of Vascular Endothelial Cells by KIAA1036 Protein To examine an effect to network formation of vascular endothelial cells by KIAA1036 protein, network formation of HUVEC in the presence (treated group) or the absence (control group) of KIAA1036 protein was observed.","Matrigel (Becton Dickinson Biosciences) (1 ml/well) was divided in 6-well tissue culture plates chilled on ice and put static at 37° C. for 1 hour to be gelatinized.","Endocell HUVEC (KURABO) were suspended in EBM medium containing 10% FCS and poured onto the matrigel at 5×104 cells/well with a final concentration of KIAA1036 protein at 1 or 10 nM (test group) or without KIAA1036 protein.","After 9 hours of incubation at 37° C., the network formations of HUVEC in test divisions and control divisions were observed with an inverted microscope.","As a result, it was confirmed that KIAA1036 protein obviously inhibits the network formation.","Industrial Applicability The present invention provides with a detection method of mRNA of KIAA1036, a detection method of a polypeptide of KIAA1036, a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or malignant neoplasm, a diagnostic kit and a therapeutic agent."]],"string":"[\n [\n \"Polypeptide serving as angiogenic marker and dna thereof\",\n \"A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease and cancer, the method of detection of the marker and a diagnosis kit of the diseases are provided.\",\n \"Additionally, therapeutic agents of the diseases are provided.\",\n \"The expression of KIAA1036 is enhanced in ovarian cancer and large bowel cancer and KIAA1036 expresses in umbilical vein endothelial cell and inhibits DNA synthesis in the cells, cell migrating and lumen formation.\",\n \"Therefore KIAA1036 is useful as a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.\",\n \"Additionally, KIAA1036 is useful for screening of agonists, antagonists, DNA synthesis inhibitors, cell migrating inhibitors and neovascular inhibitors.\",\n \"The substances obtained by the screening, KIAA1036 and the antibodies are useful as therapeutic agents the above disease.\"\n ],\n [\n \"1.A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof.\",\n \"2.A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polynucleotide hybridizing under a stringent condition with a polynucleotide having a base sequence as set forth in C of 1-C of 1480 of SEQ ID NO: 1 or fragment thereof.\",\n \"3.A marker of claim 1, which contains a polynucleotide having a base sequence as set forth in C of 1-C of 1480 of SEQ ID NO: 1 or fragment thereof.\",\n \"4.A marker of claim 1, which contains a polynucleotide having a base sequence as set forth in C of 1-C of 5481 of SEQ ID NO: 1 or fragment thereof.\",\n \"5.A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.\",\n \"6.A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polypeptide wherein 1 or more amino acid residue(s) has a mutation(s) selected from deletion, substitution and addition in an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.\",\n \"7.A marker of claim 5, which contains a polypeptide consisting of an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.\",\n \"8.A method of detection of a marker of any one of claims 1-7 from a sample derived from human.\",\n \"9.A method of detection or assay of a marker of any one of claims 1-4, which comprises the following processes and using a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof, (a) a process that the polynucleotide and a sample are contacted, and (b) a process that a polynucleotide binding with the polynucleotide is detected.\",\n \"10.A method of detection or assay of a marker of any one of claims 1-4, which comprises the following processes and using a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof, (a) a process that cDNA is prepared from a sample, and (b) a process that cDNA is amplified with the polynucleotide fragment and detected.\",\n \"11.A method of detection or assay of a marker of any one of claims 5-7, which comprises the following processes and using an antibody recognizing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (a) a process that the antibody and a sample are contacted, and (b) a process that the binding between the antibody and a polypeptide is detected.\",\n \"12.A method of detection of a disease relating to a marker of any one of claims 1-7, which comprises using a method of any one of claims 9-11.13.A method of detection of claim 12, wherein the disease is vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.\",\n \"14.A diagnostic kit of a disease relating to a marker of any one of claims 1-7, which comprises using a method of any one of claims 9-11.15.A diagnostic kit of claim 14, wherein the disease is vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.\",\n \"16.A screening method of a binding substance of a marker of any one of claims 5-7, which comprises the following processes, (a) a process that a marker of any one of claims 5-7 and a subject are contacted, and (b) a process that the binding between the marker and the subject is detected.\",\n \"17.A kit for screening of a binding substance of claim 16, which contains a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof.\",\n \"18.A screening method of a binding activity regulatory substance which regulates a binding between a marker of any one of claims 5-7 and a binding substance of the marker, which comprises the following processes, (a) a process that a marker of any one of claims 5-7 and a binding substance of the marker are contacted in the presence or absence of a subject, and (b) a process that a binding activity of the marker and the binding substance is compared in the presence or absence of a subject.\",\n \"19.A kit for screening of a binding activity regulatory substance of claim 18, which contains a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof.\",\n \"20.A screening method of an expression regulatory substance of a marker of any one of claims 1-7, which comprises the following processes, (a) a process that a cell, which can express a marker of any one of claims 1-7, and a subject are contacted, and (b) a process that a change of an expression level of the marker is detected by using a method of any one of claims 9-11.21.A kit for screening of an expression regulatory substance of a marker of any one of claims 1-7, which comprises using the method of claim 20.22.A DNA synthesis inhibitor containing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.\",\n \"23.A cell migrating activity inhibitor containing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.\",\n \"24.A pharmaceutical composition which contains at least one selected from a DNA synthesis inhibitor of claim 22, a cell migrating activity inhibitor of claim 23, a polynucleotide having a base sequence as set forth in SEQ ID NO: 1 or fragment thereof, a human gene therapeutic expression vector containing polynucleotide having a base sequence as set forth in SEQ ID NO: 1 or fragment thereof, a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof and an antibody to the polypeptide.\",\n \"25.A pharmaceutical composition of claim 24, which is a therapeutic agent for vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.\",\n \"26.A method of treating vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which comprises administrating a pharmaceutical composition of claims 24 or 25.27.Use of a polynucleotide having a base sequence as set forth in SEQ ID NO: 1 and a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 to produce a pharmaceutical composition of claim 24 or 25.\"\n ],\n [\n \"<SOH> BACKGROUND ART <EOH>By a double adapter method, shotgun library of human genome was constructed and a nucleotide sequence of AF055021 (SEQ ID NO: 3), EST clone, was determined with the DNA library (ANLITICAL BIOCHEMISTRY, 236, 107-113 (1996).).\",\n \"Subsequently, a nucleotide sequence of full-length cDNA of AF055021 (SEQ ID NO: 1) was determined and named KIAA1036 (DNA Research, 6 (3) 197-205 (1999).).\",\n \"KIAA1036 polypeptide (SEQ ID NO: 2) is a polypeptide consisting of 365 amino acids and the function has not been elucidated yet.\",\n \"Since cells need oxygen and nutriments provided from blood to survive in vivo, neovascularization is vigorous in vigorously propagating tissues.\",\n \"It is known that neovascularization relates not only to the physiological regeneration of tissues but also to diseases accompanied by the pathological cell propagation.\",\n \"As the disease, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease and cancer were reported (Nature, 407, 249 (2000).).\",\n \"Since neovascularization is also observed upon the propagation of malignant neoplasma cell, human umbilical vein endothelial cells (HUVEC) treated with vascular endothelial cell growth factor (VEGF) were reported as a model of neovascularization caused by cancer (Cancer Research, 55, 5296-5301 (1995).).\",\n \"Though the research of genes relating to the above-mentioned various diseases have been conducted, the mechanism of pathogenesis has not been perfectly elucidated, there being a possibility of unknown genes relating to the mechanism.\",\n \"Under this circumstance, an identification of a new gene relating to the pathogenesis of the above-mentioned diseases and a diagnostic procedure of diseases with the gene have been desired.\"\n ],\n [\n \"<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG.\",\n \"1 is a schematic comparing an amino acid sequence of KIAA1036 and that of AK022567.Query: 614 shows an amino acid sequence of KIAA1036.Sbjct: 1 shows an amino acid sequence of AK022567.FIG.\",\n \"2 shows an expression level of KIAA1036 in human umbilical vein endothelial cells by Northern blot.\",\n \"Total RNA was prepared by treating human umbilical vein endothelial cell with nM VEGF for 0, 0.5, 2, 6, 12 or 24 hours.\",\n \"And then signal intensity of KIAA1036 gene against signal intensity of β-actin gene was calculated.\",\n \"A relative expression level is shown when signal intensity in a control division (1 nM VEGF was not added) is 1.The expression was inhibited in 0.5 to 2 hours after VEGF was added.\",\n \"But a 3.2-fold high gene expression was induced at 24 hours after VEGF was added.\",\n \"FIG.\",\n \"3 shows an expression level of KIAA1036 in human cancer tissues by Dot blot.\",\n \"By using a nylon membrane on which cDNA was immobilized, wherein cDNA was derived from a cancer tissue obtained from a patient with human ovarian cancer or large bowel cancer and a normal tissue obtained from the same patient, signal intensity of KIAA1036 gene against signal intensity of β-actin gene was calculated.\",\n \"The ratio of signal intensity in each cancer tissue against in each normal tissue of the same patient was calculated.\",\n \"And a change of expression level of the gene in each cancer tissue was analyzed.\",\n \"As a result, in cancer tissue of all of the 4 patients with ovarian cancer and all of the 11 patients with large bowel cancer, the expression of KIAA1036 gene was increased.\",\n \"FIG.\",\n \"4 shows a cytostatic effect of KIAA1036 polypeptide against a vascular endothelial cell.\",\n \"Cell propagation in human vascular endothelial cells was calculated by measuring an uptake of bromodeoxyuridine (BrdU) in DNA synthesis.\",\n \"Human vascular endothelial cells were divided in M199 medium in which a concentration of KIAA1036 polypeptide was 0, 1 or 10 nM.\",\n \"BrdU reagent was added thereto and the cells were cultured for 12 hours.\",\n \"And then the quantity of BrdU taken in cells was measured (lane 1 to 3).\",\n \"The same operation was carried out with M199 medium to which 1 nM VEGF was added and the quantity of BrdU taken in cells was measured (lane 4 to 6).\",\n \"As a result, regardless of the addition of VEGF, DNA synthesis in cells was decreased, depending on the concentration of added KIAA1036 polypeptide.\",\n \"FIG.\",\n \"5 shows a depression effect of KIAA1036 against migrating activity of a vascular endothelial cell.\",\n \"M199 medium, in which a concentration of KIAA1036 polypeptide was 0, 1 or 10 nM, was added in the lower layer of a culture apparatus divided with a filter and vascular endothelial cells were added in the upper layer of it.\",\n \"The cells were cultured for 4 hours.\",\n \"Then cell population attached to the filter was measured (lane 1 to 3).\",\n \"The same operation was carried out with M199 medium to which 0.25 nM VEGF was added and cell population attached to the filter was measured (lane 4 to 6).\",\n \"As a result, regardless of the addition of VEGF, migrating activity of cells was inhibited, depending on the concentration of added KIAA1036 polypeptide.\",\n \"detailed-description description=\\\"Detailed Description\\\" end=\\\"lead\\\"?\"\n ],\n [\n \"TECHNICAL FIELD The present invention relates to a polypeptide and a polynucleotide useful as a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.\",\n \"And the present invention relates to the treatment of those diseases.\",\n \"BACKGROUND ART By a double adapter method, shotgun library of human genome was constructed and a nucleotide sequence of AF055021 (SEQ ID NO: 3), EST clone, was determined with the DNA library (ANLITICAL BIOCHEMISTRY, 236, 107-113 (1996).).\",\n \"Subsequently, a nucleotide sequence of full-length cDNA of AF055021 (SEQ ID NO: 1) was determined and named KIAA1036 (DNA Research, 6 (3) 197-205 (1999).).\",\n \"KIAA1036 polypeptide (SEQ ID NO: 2) is a polypeptide consisting of 365 amino acids and the function has not been elucidated yet.\",\n \"Since cells need oxygen and nutriments provided from blood to survive in vivo, neovascularization is vigorous in vigorously propagating tissues.\",\n \"It is known that neovascularization relates not only to the physiological regeneration of tissues but also to diseases accompanied by the pathological cell propagation.\",\n \"As the disease, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease and cancer were reported (Nature, 407, 249 (2000).).\",\n \"Since neovascularization is also observed upon the propagation of malignant neoplasma cell, human umbilical vein endothelial cells (HUVEC) treated with vascular endothelial cell growth factor (VEGF) were reported as a model of neovascularization caused by cancer (Cancer Research, 55, 5296-5301 (1995).).\",\n \"Though the research of genes relating to the above-mentioned various diseases have been conducted, the mechanism of pathogenesis has not been perfectly elucidated, there being a possibility of unknown genes relating to the mechanism.\",\n \"Under this circumstance, an identification of a new gene relating to the pathogenesis of the above-mentioned diseases and a diagnostic procedure of diseases with the gene have been desired.\",\n \"DISCLOSURE OF INVENTION The present inventors found that expression of KIAA1036 polypeptide was increased in ovarian cancer and colon cancer and confirmed that the peptide was useful as a marker for cancer.\",\n \"As results of further researches, they found that KIAA1036 polypeptide was a new marker for neovascular and useful as a marker and a therapeutic agent of various diseases such as vascular disease relating to neovascularization, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer to accomplish the present invention.\",\n \"The present invention is (1) a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof, (2) a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polynucleotide hybridizing under a stringent condition with a polynucleotide having a base sequence as set forth in C of 1-C of 1480 of SEQ ID NO: 1 or fragment thereof, (3) a marker of (1), which contains a polynucleotide having a base sequence as set forth in C of 1-C of 1480 of SEQ ID NO: 1 or fragment thereof, (4) a marker of (1), which contains a polynucleotide having a base sequence as set forth in C of 1-C of 5481 of SEQ ID NO: 1 or fragment thereof, (5) a marker for neovascuarlization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (6) a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which contains a polypeptide wherein 1 or more amino acid residue(s) has a mutation(s) selected from deletion, substitution and addition in an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (7) a marker of (5), which contains a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (8) a method of detection of a marker of any one of (1)-(7) from a sample derived from human, (9) a method of detection or assay of a marker of any one of (1)-(4), which comprises the following processes and using a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof, (a) a process that the polynucleotide and a sample are contacted, and (b) a process that a polynucleotide binding with the polynucleotide is detected, (10) a method of detection or assay of a marker of any one of (1)-(4), which comprises the following processes and using a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof, (a) a process that cDNA is prepared from a sample, and (b) a process that cDNA is amplified with the polynucleotide fragment and detected, (11) a method of detection or assay of a marker of any one of (5)-(7), which comprises the following processes and using an antibody recognizing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (a) a process that the antibody and a sample are contacted, and (b) a process that the binding between the antibody and a polypeptide is detected, (12) a method of detection of a disease relating to a marker of any one of (1)-(7), which comprises using a method of any one of (9)-(11), (13) a method of detection of (12), wherein the disease is vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, (14) a diagnostic kit of a disease relating to a marker of any one of (1)-(7), which comprises using a method of any one of (9)-(11), (15) a diagnostic kit of (14), wherein the disease is vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, (16) a screening method of a binding substance of a marker of any one of (5)-(7), which comprises the following processes, (a) a process that a marker of any one of (5)-(7) and a subject are contacted, and (b) a process that the binding between the marker and the subject is detected, (17) a kit for screening of a binding substance of (16), which contains a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof, (18) a screening method of a binding activity regulatory substance which regulates a binding between a marker of any one of (5)-(7) and a binding substance of the marker, which comprises the following processes, (a) a process that a marker of any one of (5)-(7) and a binding substance of the marker are contacted in the presence or absence of a subject, and (b) a process that a binding activity of the marker and the binding substance is compared in the presence or absence of a subject, (19) a kit for screening of a binding activity regulatory substance of (18), which contains a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof, (20) a screening method of an expression regulatory substance of a marker of any one of (1)-(7), which comprises the following processes, (a) a process that a cell, which can express a marker of any one of (1)-(7), and a subject are contacted, and (b) a process that a change of an expression level of the marker is detected by using a method of any one of (9)-(11), (21) a kit for screening of an expression regulatory substance of a marker of any one of (1)-(7), which comprises using the method of (20), (22) a DNA synthesis inhibitor containing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (23) a cell migrating activity inhibitor containing a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof, (24) a pharmaceutical composition which contains at least one selected from a DNA synthesis inhibitor of (22), a cell migrating activity inhibitor of (23), a polynucleotide having a base sequence as set forth in SEQ ID NO: 1 or fragment thereof, a human gene therapeutic expression vector containing polynucleotide having a base sequence as set forth in SEQ ID NO: 1 or fragment thereof, a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 or fragment thereof and an antibody to the polypeptide, (25) a pharmaceutical composition of (24), which is a therapeutic agent for vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, (26) a method of treating vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, which comprises administrating a pharmaceutical composition of (24) or (25), (27) use of a polynucleotide having a base sequence as set forth in SEQ ID NO: 1 and a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2 to produce a pharmaceutical composition of (24) or (25).\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is a schematic comparing an amino acid sequence of KIAA1036 and that of AK022567.Query: 614 shows an amino acid sequence of KIAA1036.Sbjct: 1 shows an amino acid sequence of AK022567.FIG.\",\n \"2 shows an expression level of KIAA1036 in human umbilical vein endothelial cells by Northern blot.\",\n \"Total RNA was prepared by treating human umbilical vein endothelial cell with nM VEGF for 0, 0.5, 2, 6, 12 or 24 hours.\",\n \"And then signal intensity of KIAA1036 gene against signal intensity of β-actin gene was calculated.\",\n \"A relative expression level is shown when signal intensity in a control division (1 nM VEGF was not added) is 1.The expression was inhibited in 0.5 to 2 hours after VEGF was added.\",\n \"But a 3.2-fold high gene expression was induced at 24 hours after VEGF was added.\",\n \"FIG.\",\n \"3 shows an expression level of KIAA1036 in human cancer tissues by Dot blot.\",\n \"By using a nylon membrane on which cDNA was immobilized, wherein cDNA was derived from a cancer tissue obtained from a patient with human ovarian cancer or large bowel cancer and a normal tissue obtained from the same patient, signal intensity of KIAA1036 gene against signal intensity of β-actin gene was calculated.\",\n \"The ratio of signal intensity in each cancer tissue against in each normal tissue of the same patient was calculated.\",\n \"And a change of expression level of the gene in each cancer tissue was analyzed.\",\n \"As a result, in cancer tissue of all of the 4 patients with ovarian cancer and all of the 11 patients with large bowel cancer, the expression of KIAA1036 gene was increased.\",\n \"FIG.\",\n \"4 shows a cytostatic effect of KIAA1036 polypeptide against a vascular endothelial cell.\",\n \"Cell propagation in human vascular endothelial cells was calculated by measuring an uptake of bromodeoxyuridine (BrdU) in DNA synthesis.\",\n \"Human vascular endothelial cells were divided in M199 medium in which a concentration of KIAA1036 polypeptide was 0, 1 or 10 nM.\",\n \"BrdU reagent was added thereto and the cells were cultured for 12 hours.\",\n \"And then the quantity of BrdU taken in cells was measured (lane 1 to 3).\",\n \"The same operation was carried out with M199 medium to which 1 nM VEGF was added and the quantity of BrdU taken in cells was measured (lane 4 to 6).\",\n \"As a result, regardless of the addition of VEGF, DNA synthesis in cells was decreased, depending on the concentration of added KIAA1036 polypeptide.\",\n \"FIG.\",\n \"5 shows a depression effect of KIAA1036 against migrating activity of a vascular endothelial cell.\",\n \"M199 medium, in which a concentration of KIAA1036 polypeptide was 0, 1 or 10 nM, was added in the lower layer of a culture apparatus divided with a filter and vascular endothelial cells were added in the upper layer of it.\",\n \"The cells were cultured for 4 hours.\",\n \"Then cell population attached to the filter was measured (lane 1 to 3).\",\n \"The same operation was carried out with M199 medium to which 0.25 nM VEGF was added and cell population attached to the filter was measured (lane 4 to 6).\",\n \"As a result, regardless of the addition of VEGF, migrating activity of cells was inhibited, depending on the concentration of added KIAA1036 polypeptide.\",\n \"BEST MODE FOR CARRYING OUT THE INVENTION Preparation of KIAA1036 polynucleotide which is a marker of the present invention, preparation of the polypeptide, a recombinant vector, a transformant, a method of production thereof, an antibody, the method of detection of polypeptide, a method of detection of mRNA, the method of detection of neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer and a diagnostic kit, a therapeutic agent or a method of treating for vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer are explained below.\",\n \"In this description, if there is no instruction, a gene recombinant technique, a production engineering technique of a recombinant polypeptide in animal cells, insect cells, yeast and Escherichia coli, a molecular-biological method, the method of separation and purification of expressed polypeptide, assay and immunological method, which are well-known in this field, are adopted.\",\n \"A marker of the present invention is “a polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 or fragment thereof” and a polynucleotide sequence of KIAA1036.Additionally, a marker of the present invention includes “a polynucleotide having a base sequence as set forth in C of 1-C of 1480 of SEQ ID NO: 1 or fragment thereof” and “a polynucleotide hybridizing with the present invention marker under a stringent condition or fragment thereof”.\",\n \"Hereinafter, a polynucleotide, which is a marker of the present invention, is referred to as polynucleotide A.\",\n \"In this description, “neovascularization” means a phenomenon that a new vasculature is developed from an existing vasculature.\",\n \"A disease related to “neovascular” includes “vascular disease”, “inflammatory disease”, “entoptic neovascular disease”, “reproductive system disease”, “central nervous system disease” or “cancer”.\",\n \"“Vascular disease” includes arterial sclerosis, hypertonia, angina pectoris, obstructive arteriosclerosis, myocardial infarction, cerebral infarction, diabetic angiopathy or vascular malformation.\",\n \"“Inflammatory disease” includes hepatitis, pneumonitis, glomerular nephritis, thyreoiditis, osteitis, arthromeningitis, osteoclasia, chondrolysis, rheumatism, bronchial asthma, sarcoidosis, Crow-Fukase syndrome, pannus, allergic oedema, ulcers, hydroperitoneum, peritoneal screlosis or tissular conglutination.\",\n \"“Entoptic neovascular disease” includes diabetic retinopathy, occlusion of retinal vein or aging macular degeneration.\",\n \"“Reproductive system disease” includes uterus dysfunction, placental dysfunction, ovarian hyperergasia or follicle cyst.\",\n \"“Central nervous system disease” includes retinosis, cerebral apoplexy, vascular dementia or Alzheimer disease.\",\n \"“Cancer” includes a malignant neoplasma such as solid cancer, angiomatous, hemangioendothelioma, sarcomas, sarcoma or hematopoietic organic ulcer, ovarian cancer or large bowel cancer.\",\n \"Additionally, it includes metastatics of these cancers.\",\n \"“A polynucleotide hybridizing under a stringent condition” means a polynucleotide obtainable by a generally known and common method in this field with using a fragment of polynucleotide A as a probe, for example, colony hybridization, plaque hybridization or Southern blotting hybridization.\",\n \"An example of the above polynucleotide includes a polynucleotide obtained by hybridization at 65° C. in the presence of 0.7 to 1.0M NaCl by using a membrane on which polynucleotides from colony or plaque are immobilized and a washing of a membrane at 65° C. with SSC (Saline Sodium Citrate; 150 mM sodium chloride, 15 mM sodium citrate) solution whose concentration is 0.1-fold to twice.\",\n \"Hybridization can be performed as well as methods described in Molecular Cloning: A Laboratory Manual, Second Edition (1989)(Cold Spring Harbor Laboratory Press), Current Protocols in Molecular Biology (1994)(Wiley-Interscience), DNA Cloning 1: Core Techniques, A practical Approach, Second Edition (1995)(Oxford University Press).\",\n \"Preferably, sequences comprising of only adenine (a) or thymine (T) are excluded from sequences hybridizing under a stringent condition.\",\n \"“A fragment of a polynucleotide” includes a polynucleotide having a base sequence comprising 5 or more serial bases in a base sequence of SEQ ID NO: 1, for example, a polynucleotide having a base sequence comprising 5, 8, 10, 12, 15, 20, 25, 30, 50, 100, 500 or 1000 bases.\",\n \"In this description, “a polynucleotide hybridizing” includes a polynucleotide hybridizing with the other polynucleotide under the above hybridizing condition.\",\n \"Examples of the above polynucleotide includes a polynucleotide having a homology of at least 60% or more, preferably 80% or more and more preferably 95% or more with a base sequence of DNA encoding a polypeptide having an amino acid sequence of SEQ ID NO: 1.And the homology is shown as a score, for example, by using a search program BLAST with algorithm developing by Altschul et al (The Journal of Molecular Biology, 215, 403-410 (1990).).\",\n \"A marker of the present invention includes “a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof”, a polypeptide sequence of KIAA1036 including their salts.\",\n \"A “salt” includes a salt physiologically acceptable with an acid or a base.\",\n \"A salt with an acid is especially preferable.\",\n \"The salt includes, for example, a salt with inorganic acid such as hydrochloric acid, phosphoric acid, hydrobromic acid or sulfuric acid, or a salt with organic acid such as acetic acid, fomic acid, proprionic acid, fumaric acid, maleic acid, succinic acid, tartaric acid, citric acid, malic acid, oxalic acid, benzoic acid, methanesulfonic acid or benzenesulfonic acid.\",\n \"“A fragment of polypeptide” includes a polypeptide sequence comprising 5 or more serial amino acid residues in an amino acid sequence of SEQ ID NO: 2, for example, a polypeptide comprising an amino acid sequences comprising 5, 8, 10, 12, 15, 20, 25, 30, 50 or 100 amino acid residues.\",\n \"A marker of the present invention includes “a polypeptide having a mutation selected from deletion, substitution and addition in 1 or a few amino acid residue(s) of an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof”.\",\n \"Hereinafter, a polypeptide, which is a marker of the present invention, is referred to as polypeptide B.\",\n \"“1 or a few amino acid residue(s)” means amino acid residues, which can delete, substitute or add by a site-specific mutagenic method or the like.\",\n \"Namely, it means amino acid residues having 50 or less, preferably 30 or less, more preferably 20 or less, much more preferably 10 or less amino acid residues.\",\n \"Furthermore, polypeptide B includes a polypeptide to be used as a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer even after deletions, substitutions or additions.\",\n \"It is, for example, a polypeptide having a homology of at least 60% or more, preferably 80% or more and more preferably 95% or more with an amino acid sequence of SEQ ID NO: 2.In this description, “a marker” includes a substance used to detect neovascularization and all kinds of disease related to neovascularization derived from organism samples, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.\",\n \"And it includes polynucleotide A and polypeptide B. mRNA or a protein can be a “marker” when the expression increases or decreases under diseases.\",\n \"Examples of the above maker include KIAA1036 polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2, KIAA1036 polynucleotide having a base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1, a polynucleotide having a complementary base sequence thereof or a fragment thereof.\",\n \"In this description, terms are used with usual meanings in this field unless otherwise mentioned especially referred.\",\n \"The terms especially used in this description are explained below.\",\n \"An “antibody” means a general antibody in this field and includes all of the antibodies, fragments thereof, derivatives, conjugations or modified antibodies.\",\n \"An antibody recognizing polypeptide B or fragment thereof is preferable, an antibody recognizing specifically the above polypeptide is more preferable and an antibody recognizing single specifically the polypeptide is much more preferable.\",\n \"These antibodies include a polyclonal antibody or a monoclonal antibody.\",\n \"“A detection or a quantitation” of a marker of the present invention can be accomplished by any appropriate method including an immunological assay or a molecular-biological assay.\",\n \"When a marker is polypeptide B, the above method includes, for example, an immunological assay such as ELISA (Enzyme Linked Immuno Sorbent Assay), RIA (Radio Immuno Assay), fluorescence antibody technique, Western blot or an immune structure dyeing method.\",\n \"By using an antibody against polypeptide B or fragment thereof, neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer can be detected or quantitated.\",\n \"Because a marker of the present invention can be used as a marker for above diseases, a method of detection or quantitation of diseases with the above antibody can be accomplished by any appropriate method including an immunological assay as well as an above detection or quantitation of a polypeptide.\",\n \"The present invention relates to a diagnostic kit of neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, characterized by including an antibody against polypeptide B or fragment thereof.\",\n \"The above kit includes at least an antibody against polypeptide B or fragment thereof and a standard reagent of polypeptide B.\",\n \"Because polypeptide B can be used as a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, it can be immunologically measured as well as an above mentioned method of detection or quantitation by an immunological assay.\",\n \"And when a marker is a polynucleotide such as mRNA, the assay includes a molecular-biological assay, for example, Northern blot, Dot blot or polymerase chain reaction (PCR).\",\n \"mRNA can be detected or quantitated by using polynucleotide A or fragment thereof as a probe or primer.\",\n \"Because a marker of the present invention can be a marker of above diseases, a method of detection or quantitation of diseases with the probe or the primer is accomplished by an appropriate method including a molecular-biological assay as well as the above method for detection or quantitation of a polynucleotide.\",\n \"A probe or a primer includes a nucleotide comprising serial bases in a base sequence of SEQ ID NO: 1, for example, a base sequence comprising 5, 8, 10, 12, 15, 20, 25, 30, 40, 50 or 60 bases.\",\n \"It includes an oligonucleotide having the same sequence, an oligonucleotide having a sequence complementary to the sequence of the oligonucleotide, and the derivative thereof.\",\n \"A derivative thereof includes, for example, a oligonucleotide wherein a phosphodiester bond in the oligonucleotide is transformed into a phosphorothioate bond or a N3′-P5′ phosphoamidite bond, a oligonucleotide wherein a ribose and a phosphodiester bond are transformed into a peptide bond, a oligonucleotide wherein a uracil in the oligonucleotide is substituted with a C-5 propionyl uracil or a C-5 thiazole uracil, a oligonucleotide wherein a cytosine in the oligonucleotide is substituted with C-5 propionyl cytosine or cytosine modified with phenoxazine or a oligonucleotide wherein a ribose in DNA is substituted with 2′-O-propyl ribose, 2′-methoxyethoxy ribose or the like.\",\n \"Those polynucleotides are useful, for example, as a gene marker, a primer for PCR or a probe for hybridization.\",\n \"The present invention relates to a part or all of the polynucleotide encoding polypeptide B and a diagnostic kit of vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer including the polynucleotide as a standard reagent.\",\n \"This kit includes at least a probe or a primer against polynucleotide A or fragment thereof and a standard reagent of polynucleotide A.\",\n \"Because polynucleotide A can be used as a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, it is molecular-biologically measured as well as the above method of detection or quantitation by a molecular-biological assay.\",\n \"A transcription of DNA or a translation of mRNA can be inhibited by using polynucleotide A.\",\n \"Methods to inhibit transcription of DNA or translation of mRNA of the polypeptide B can be provided by appropriately preparing a polynucleotide having a complementary base sequence to a base sequence of polynucleotide A, a polynucleotide hybridizing with polynucleotide A or an oligonucleotide and a derivative oligonucleotide having a complementary sequence to an oligonucleotide having serial 5 to 60 base sequences in base sequence of polynucleotide A and using antisense RNA/DNA techniques known in this field.\",\n \"The present invention relates to “a screening method of a binding substance” of a marker of the present invention.\",\n \"“A binding substance” includes any substance binding to a marker of the present invention.\",\n \"The binding substance includes, for example, a low-molecular weight substance, a polynucleotide or a polypeptide such as a receptor and an antibody.\",\n \"“A screening method of a binding substance” can be accomplished by appropriately using techniques widely known in this technical field.\",\n \"It includes, for example, a bioassay and a binding assay.\",\n \"“A kit for screening of a binding substance” of the present invention includes at least a marker of the present invention.\",\n \"A binding substance can be screened by biochemical method with the above kit.\",\n \"Additionally the present invention relates to “a screening method of a binding activity regulatory substance”.\",\n \"“A binding activity regulation” means that a binding activity of a marker of the present invention and the binding substance is enhanced or reduced and “a binding activity” is evaluated by Percent Maximum Binding (PMB).\",\n \"“A binding activity regulatory substance” includes a substance that makes a binding activity of the binding substance against a marker of the present invention enhanced or reduced, for example, an agonist or an antagonist.\",\n \"The substance includes, for example, a low-molecular weight substance, a polynucleotide or a polypeptide.\",\n \"“A screening method of a binding activity regulatory substance” can be accomplished by appropriately using techniques widely known in this field.\",\n \"A binding activity regulatory substance can be screened by a screening method of the present invention.\",\n \"The method is, for example, a bioassay or a binding assay.\",\n \"“A screening kit of a binding activity regulatory substance” of the present invention includes at least a marker of the present invention.\",\n \"A binding activity regulatory substance can be screened by a biochemical method with a kit of the present invention.\",\n \"And the present invention relates to “a screening method of an expression regulatory substance”.\",\n \"Expression level means the amount of a marker of the present invention expressed in target cells.\",\n \"Expression level can be evaluated by an appropriate method including an immunological assay with an antibody against polypeptide B, for example, ELISA, RIA, a fluorescence antibody technique or an immune structure dyeing method or by a molecular-biological assay measuring mRNA of polynucleotide A, for example, Northern blot hybridization, Dot blot or RT-PCR.\",\n \"“Expression regulation” means that expression level of a marker of the present invention in protein or mRNA level evaluated by any appropriate method including the above immunological or molecular-biological assay is enhanced or reduced.\",\n \"“An expression regulatory substance” includes a substance enhancing or reducing expression level of a marker of the present invention in protein or mRNA level, which can be evaluated by any appropriate method including the above immunological or molecular-biological assay.\",\n \"The above substance includes, for example, a low-molecular weight substance, a polynucleotide or a polypeptide.\",\n \"“A screening method of an expression regulatory substance” can be accomplished by appropriately using techniques widely known in this field.\",\n \"An expression regulatory substance can be screened by a screening method of the present invention.\",\n \"The method includes, for example, a bioassay or a binding assay.\",\n \"“A screening kit of an expression regulatory substance” of the present invention includes at least a marker of the present invention.\",\n \"An expression regulatory substance can be screened by a biochemical method based on a screening method of an expression regulatory substance with a kit of the present invention.\",\n \"Because polypeptide B inhibits a DNA synthesis in a vascular endothelial cell and a migrating activity of a vascular endothelial cell, polynucleotide A, a complementary polynucleotide of the above polynucleotide, polypeptide B or/and an antibody against the above polypeptide can be a DNA synthesis inhibitor or/and a cell migrating activity inhibitor.\",\n \"Then, the present invention provides a DNA synthesis inhibitor and a cell migrating inhibitor.\",\n \"“A pharmaceutical composition” of the present invention includes at least one selected from polypeptide B, an antibody against the polypeptide, polynucleotide A, a complementary polynucleotide of the polynucleotide, a DNA synthesis inhibitor, a cell migrating activity inhibitor, a binding substance, a binding activity regulatory substance and an expression regulatory substance.\",\n \"It can be used for a therapeutic agent of a specific disease, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer.\",\n \"The substance includes, for example, a low-molecular weight substance, a polynucleotide or a polypeptide.\",\n \"Preferred is a polypeptide comprising an amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 or fragment thereof.\",\n \"And a pharmaceutical composition of the present invention includes “a gene therapeutic expression vector”.\",\n \"“A gene therapeutic expression vector” includes an expression vector inserted all or a part of polynucleotide A and a vector which inserts a normal gene in a cell, repairs/modifies a defect of a gene and regulates an expression of a gene by introducing a gene in a cell/tissue.\",\n \"The vector includes a viral vector wherein a part or all of a base sequence of a viral, which is lack of reproductive capability, is replaced with a therapeutic gene.\",\n \"Then, the present invention provides therapeutic agents or methods of treating for these diseases.\",\n \"“A method of treating” includes a method for treating of specific diseases, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer relating to neovascularization by a administration of the above pharmaceutical composition.\",\n \"Hereinafter, the Present Invention is Described in Detail.\",\n \"(1) An Preparation of KIAA1036 Polynucleotide cDNA library is manufactured from human brain, heart, skeletal muscle, spleen, kidney, liver, small intestine, placenta, human normal cells from these tissues, human umbilical vein endothelial cells or human ovarian cancer or human large bowel cancer by a usual method.\",\n \"A method which manufactures cDNA library is a method described in Molecular Cloning: A Laboratory Manual, Second Edition (1989) (Cold Spring Harbor Laboratory Press), Current Protocols in Molecular Biology (1994) (Wiley-Interscience) or DNA Cloning 1: Core Techniques, A Practical Approach, Second Edition (1995) (Oxford University Press) or the method that a kit on the market, for example, SuperScript Plasmid System for cDNA Synthesis and Plasmid Cloning (Invitrogen) or ZAP-cDNA Synthesis Kits (STRATAGENE) is used.\",\n \"cDNA obtained by the method is, for example, DNA encoding a polypeptide of SEQ ID NO: 2.Examples of the above cDNA include a DNA comprising a base sequence of SEQ ID NO: 1.A plasmid including DNA of SEQ ID NO: 1 is, for example, a plasmid described in the following examples.\",\n \"The obtained DNA is inserted to an appropriate expression vector and an expression plasmid is constructed.\",\n \"The expression vectors are introduced to appropriate hosts and then transformants can be obtained.\",\n \"The expression vector is any vector in which cDNA is inserted and which express in animal cells.\",\n \"The vector is, for example, pcDNA1.1, pcDNA1.1/Amp, pCDM8, pREP (Invitrogen), pHM6, pHB6 (Roche Diagnostics), pKK223-3, pGEX (Amersham Pharmacia Bioteque), pET-3, pET-11, pBluescriptII SK(+), pBluescriptII SK(−) (STRATAGENE), pUC19, pTrxFus (Invitrogen), pUC118, pSTV28 (TaKaRa), pMAL-c2X (New England BioLabs), pAGE107 (Cytotechnology, 3 (2), 133-140 (1990).\",\n \"; JP1991-22979), pAGE103 (The Journal of Biochemistry, 101 (5), 1307-1310 (1987).\",\n \"), pAMo, pAMoA (The Journal of Biological Chemistry, 268 (30), 22782-22787 (1993).\",\n \"), pAMoPRSA (JP1993-336963) or pAS3-3 (JP1990-227075).\",\n \"The expression vectors with a cDNA insert introduce a target cDNA into optional animal cells and transformed cells are obtained.\",\n \"Any method for introducing DNA can be used as a method for introducing the expression vectors a host.\",\n \"When a host is animal cells, it is, for example, an electroporation (Cytotechnology, 3 (2), 133-140 (1990).\",\n \"), a calcium phosphate method (JP1990-227075) or a lipofection method (Proceedings of the National Academy of Sciences USA, 84, 7413 (1987).\",\n \"; Virology, 52, 456 (1973).).\",\n \"Appropriate cells or tissue for expression vectors can be used as a host.\",\n \"It is, for example, animal cells.\",\n \"An animal cell as a host is, for example, Namalwa (Burkitt lymphoma, ATCC: CRL-1432) and the subline NamalwaKJM-1 which is an established cell from human, HCT-15 (human large bowel cancer cell, ATCC: CCL-225), COS-1(African green monkey's nephrocyte (SV40 transformed cell), ATCC: CRL-1650) and COS-7 (African green monkey's nephrocyte (SV40 transformed cell), ATCC: CRL-1651) which is an established cell from monkey or CHO-K1 (Chinese hamster ovary cell, ATCC: CCL-61) and HBT5637 (JP1987-299) which is an established cell derived from hamster.\",\n \"Preferred is Namalwa cell, NamalwaKJM-1 cell or HCT-15 cell.\",\n \"Transformants of the present invention are cultured by a generally known and common method in this field.\",\n \"It can be accomplished with a medium appropriate to a transforming host and a liquid medium is appropriate as a medium for a culturing liquid medium.\",\n \"For example, MEM medium (Science, 130, 432 (1959).\",\n \"), D-MEM medium (Virology, 8, 396 (1959).\",\n \"), PRMI 1640 medium (The Journal of the American Medical Association, 199, 519 (1967).\",\n \"), YT medium or BEM medium can be used.\",\n \"When transformants is prepared by using animal cells as a host are cultured, for example, a medium wherein cattle fetal serum (FCS) is appropriately added into MEM medium, D-MEM medium or PRIM medium can be used.\",\n \"A medium can optionally include a substance promoting transcription activity to enhance transcription activity of a promoter of an expression vector.\",\n \"For example, isopropyl-1-thio-β-D-galactopyranosin (IPTG) can be used.\",\n \"A medium includes nutriments needed to grow up transformants, for example, glucose, amino acid, peptone, vitamin, hormone or serum, preferably, FCS, calcium chloride or magnesium chloride.\",\n \"The medium like this, which has any composition, can be used and a medium available on the market can be used.\",\n \"A cultivation is held in pH 6.0 to 8.0 at 25 to 40° C. in the presence of 5% CO2.The obtained transformed cells are cultured by a usual method.\",\n \"For example, they can be cultured by the following culture method.\",\n \"When transformants are animal cells, examples of a medium for culturing the cells includes commonly used RPMI1640 medium (The Journal of the American Medical Association, 199, 519 (1967).\",\n \"), MEM medium of Eagle (Science, 122, 501 (1952).\",\n \"), D-MEM medium (Virology, 8, 396 (1959).\",\n \"), 199 medium (Proceeding of the Society for the Biological Medicine, 73, 1 (1950).)\",\n \"or a medium added fetal calf serum (FCS).\",\n \"Cultivation is usually carried out in pH 6 to 8, at 25 to 40° C., in the presence of 5% CO2 for 1 to 7 days.\",\n \"An antibiotic such as kanamycin, penicillin or streptomycin can optionally be added during cultivation.\",\n \"DNAs encoding KIAA1036 which is a neovascular marker can be obtained from human brain, heart, skeletal muscle, spleen, kidney, liver, lung, placenta, human normal cells from these tissues, human umbilical vein endothelial cells or human ovarian cancer or human large bowel cancer by the above-mentioned.\",\n \"On the basis of an amino acid sequence, DNA encoding KIAA1036 can be prepared by a chemical synthesis.\",\n \"A chemical synthesis of DNA can be accomplished with a DNA synthesizer using a thiophosphite method (Shimazu Corporation) or a DNA synthesizer model 392 using a phosphoamidite method (PerkinElmer, Inc.).\",\n \"The following oligonucleotide is used as a sense primer (SEQ ID NO: 4) or an antisense primer (SEQ ID NO: 5) and cDNA prepared from mRNA in cells expressing complementary mRNA of these DNA is used as a template.\",\n \"Target DNA can be prepared by PCR under this condition.\",\n \"(2) The Production of KIAA1036 Polypeptide KIAA1036 can be produced by a method described in Molecular Cloning: A Laboratory Manual, Second Edition (1989)(Cold Spring Harbor Laboratory Press) or Current Protocols in Molecular Biology (1994)(Wiley-InterScience).\",\n \"For example, KIAA1036 polynucleotide is made to express in host cells by the following method and KIAA1036 can be produced.\",\n \"On a basis of a full length of DNA encoding KIAA1036 polypeptide, an appropriate length of DNA fragment including a part encoding the polypeptide is optionally prepared.\",\n \"And DNA whose bases substituted is prepared, as a base sequence encoding the polypeptide is the most suitable codons for expression in a host.\",\n \"This DNA is useful to enhance productivity of the polypeptide.\",\n \"By inserting the DNA fragment or a full length of DNA to the downstream of a promoter of an appropriate expression vector, a recombinant DNA (a recombinant vector) is produced.\",\n \"By the recombinant vectors are produced in host cells adapting to the expression vector, transformants producing KIAA1036 polypeptide can be obtained.\",\n \"Any cell such as a prokaryotic cell, yeast, an animal cell, a plant cell or a insect cell can be used as a host cell, if a target gene can be expressed in the cells.\",\n \"As an expression vector, the vector which can autonomously replicate in the above host cells or be inserted into a chromosome and which includes a promoter at the appreciate position for the transcription of KIAA1036 polypeptide gene.\",\n \"(i) The case that prokaryote is used as a host.\",\n \"An expression vector of KIAA1036 can autonomously replicate in prokaryote as well as is preferably constructed with a promoter, a ribosome binding sequence, KIAA1036 and a transcription termination sequence.\",\n \"A gene regulating a promoter can be included.\",\n \"An expression vector is, for example, pBTrp2, pBTac1, pBTac2 (Roche Diagnostics), BluescriptII SK(+), pBluescriptII SK(−) (STRATAGENE), pSTV28, pUC118, pUC19 (TaKaRa), pKK233-2 (Pharmacia), pSE280, pSupex, pUB110, pTP5, pC194, pTrxFus (Invitrogen), pGEMEX-1 (Promega), pQE-8 (QIAGEN), pGEX (Pharmacia), pETsystem (Novagen), pMAL-c2 (New England BioLabs), pKYP10 (JP1982-110600), pKYP200 (Agricultural Biological Chemistry, 48, 669 (1984).\",\n \"), pLSA1 (Agricultural Biological Chemistry, 53, 277 (1989).\",\n \"), pGEL1 (Proceedings of the National Academy of Sciences USA, 82, 4306(1985).\",\n \"), pEG400 (Journal of Bacteriology, 172, 2392(1990).\",\n \"), pTrs30 (FERM BP-5407), pTrs32 (FERM BP-5408), pGHA2 (FERM BP-400), pGKA2 (FERM BP-6798), pPA1 (JP1987-233798) or pTerm2 (JP1990-22979, U.S. Pat.\",\n \"No.\",\n \"4,686,191, U.S. Pat.\",\n \"No.\",\n \"4,939,094, U.S. Pat.\",\n \"No.\",\n \"5,160,735).\",\n \"Any promoter which can express in a host cell such as Escherichia coli can be used.\",\n \"For example, it is a promoter from Escherichia coli or a phage such as trp promoter (Ptrp), lac promoter (Plac), PL promoter, PR promoter or PSE promoter, SPO1 promoter, SPO2 promoter or penP promoter.\",\n \"And a promoter artificially modified such as a promoter which two Ptrps is connected in series (Ptrp×2), tac promoter, lacT7 promoter or letI promoter can be used.\",\n \"Preferred is a plasmid that the distance between Shine-Dalgarno sequence which is a ribosome binding sequence and an initiation codon is regulated to an appropriate distance, for example, 6 to 18 bases.\",\n \"A transcription termination sequence is not always needed for an expression of KIAA1036 polynucleotide but preferred is that a transcription termination sequence is positioned at the right downstream of a structural gene.\",\n \"A host cell is, for example, a prokaryote of Escherichia genus, Serratia genus, Bacillus genus, Brevibacterium genus, Corynebacterium genus, Microbacterium genus or Pseudomonas genus, for example, XL1-Blue strain, XL2-Blue strain, DH1 strain, MC1000 strain, KY3276 strain, W1485 strain, JM109 strain, HB101 strain, No.\",\n \"49 strain, W3110 strain, NY49 strain, BL21 (DE3) strain, BL21 (DE3) pLysS strain, HMS174 (DE3) strain or HMS174 (DE3) pLysS strain of E. coli as Escherichia genus, S. ficaria strain, S. fonticola strain, S. liquefaciens strain or S. marcescens strain as Serratia genus, B. subtilis strain or B. amyloliquefaciens strain as Bacillus genus, B. ammoniagenes strain, B. Immariophilum (ATCC: 14068) strain or B. saccharolyticum (ATCC: 14066) strain as Brevibacterium genus, C. glutamicum (ATCC: 13032) strain, C. glutamicum (ATCC: 14067) strain, C. glutamicum (ATCC: 13869) strain or C. acetoacidophilum (ATCC: 13870) strain as Corynebacterium genus, M. ammoniaphilum (ATCC: 15354) strain as Microbacterium genus or S. mephitica strain as Pseudomonas genus.\",\n \"Any method to introduce DNA to the above host cell can be used as an introduction method of a recombinant vector, for example, an electroporation (Nucleic Acids Research, 16, 6127 (1988).\",\n \"), calcium phosphate method (Proceedings of the National Academy of Sciences USA, 69, 2110 (1972).\",\n \"), a protoplast method (JP1987-2483942) or the method described in Gene, 17, 107 (1982), or Molecular & General Genetics, 168, 111 (1979).\",\n \"(ii) The Case that Yeast is Used as a Host.\",\n \"When yeast is used as a host, an expression vector is, for example, YEp13 (ATCC:37115), YEp24 (ATCC:37051), YCp50 (ATCC:37419), pHS19 or pHS15.Any promoter which express in yeast can be used as a promoter, for example, ADH1 (alcohol dehydrogenase) promoter, PHO5 (acid phosphatase) promoter, PGK1 (phosphoglycerate kinase) promoter, GAPDH (glyceraldehyde3-phosphate dehydrogenase) promoter, GAL1 (galactose kinase) promoter, GAL10 (UDP galactose 4-epimerase) promoter, MF α 1 (α pheromone) promoter or CUP1 (metallothionein) promoter.\",\n \"A host is, for example, S. cerevisiae species of Saccharomyces genus, S. pombe species of Schizosaccharomyces genus, K. lactis species of Kluyveromyces genus, T. pullulans species of Trichosporon genus, S. alluvius species of Schwanniomyces genus or P. pastoris species of Pichia genus.\",\n \"Any method to introduce DNA into a host can be used as a introduction method of a recombinant vector, for example, an electroporation (Methods in Enzymology, 194, 182 (1990).\",\n \"), a spheroplast method (Proceedings of the National Academy of Sciences USA, 84, 1929 (1978).)\",\n \"or a lithium acetate method (Journal of Bacteriology, 153, 163 (1983), or Proceedings of the National Academy of Sciences USA, 75, 1929 (1978).).\",\n \"(iii) The Case that an Animal Cell is Used as a Host.\",\n \"When an animal cell is used as a host, an expression vector is, for example, pcDNA1/Amp, pcDNA1, pCDM8, pREP4 (Invitrogen), pAGE 107 (Cytotechnology, 3, 133 (1990).\",\n \"), pAGE 103 (The Journal of Biochemistry, 101, 1307 (1987).\",\n \"), pAMo, pAMoA (pAMoPRSA) (The Journal of Biological Chemistry, 268, 22782-22787 (1993).)\",\n \"or pAS3-3 (JP1990-22705).\",\n \"Any promoter that can express in a host can be used as a promoter, for example, a promoter of IE (Imediate-early) gene of human cytomegalovirus (hCMV), an early promoter of SV40, Long Terminal Repeat Promoter of Moloney Murine Leulemia Virus, a promoter of retrovirus, HSP promoter, SR α promoter or a promoter of metallothionein.\",\n \"An enhancer of IE gene of human CMV can be used with a promoter.\",\n \"An animal cell as a host is, for example, HEK293 (a human fetal nephrocyte, ATCC:CRL-1573), Namalwa (Burkitt lymphoma, ATCC:CRL-1432), HeLa (a cell of carcinoma of uterine cervix, ATCC:CCL-2), HBT5637 (a leukemia cell, JP1987-299), BALL-1 (a leukemia cell) or HCT-15 (a large bowel cancer cell) of an established cell from a human, Sp2/0-Ag14 (a mouse myeloma cell, ATCC:CRL-1581) or NSO (a mouse myeloma cell) of an established cell from a mouse, COS-1 (African green monkey nephrocyte (SV40 transformed cell), ATCC:CRL-1650) or COS-7 (African green monkey nephrocyte (SV40 transformed cell), ATCC:CRL-1651) of an established cell from a monkey, CHO-K1 (Chinese hamster ovary cell, ATCC:CCL-61) or BHK-21 (C-13) (Sicilian hamster kidney cell, ATCC:CCL-10) of an established cell from a hamster, PC12 (an adrenal pheochromocytoma, ATCC:CRL-1721) or YB2/0 (a rat myeloma cell, ATCC:CRL-1662) of an established cell from a rat.\",\n \"Any method to introduce DNA into a host can be used as an introduction method of a recombinant vector, for example, an electroporation (Cytotechnology, 3, 133, (1990).\",\n \"), a calcium phosphate method (JP1990-22705) or a lipofection method (Proceedings of the National Academy of Sciences, USA, 84, 7413 (1987), or Virology, 52, 456 (1973).).\",\n \"(iv) The Case that a Plant Cell is Used as a Host.\",\n \"When a plant cell or a plant is used as a host, a polypeptide can be produced as well as a well-known method (The Tissue Culture, 20 (1994)., The Tissue Culture, 21 (1995), or Trends in Biotechnology, 15, 45 (1997).).\",\n \"An expression vector is, for example, Ti plasmid or Tobacco mosaic virus vector.\",\n \"Any promoter that can express in a plant cell can be used as a promoter for a gene expression, for example, 35S promoter of Cauliflower mosaic virus (CaMV) or Rice actin 1 promoter.\",\n \"And expression productivity of a gene can be enhanced by inserting intron 1 of an alcohol dehydrogenase gene of maize between a promoter and an expressed.\",\n \"A host is, for example, a plant cell such as potato, tobacco, maize, rice, rape, soybean, tomato, carrot, wheat, barley, rye, alfalfa or flax.\",\n \"Any method to introduce DNA into a host can be used as a introduction method of a recombinant vector, for example, a method with Agrobacterium (JP1983-140885, JP1984-70080 or WO94/00977), an electroporation (JP1984-251887) or a particle gun (gene gun) method (JP2606856, JP2517813).\",\n \"(v) The Case that an Insect Cell is Used as a Host.\",\n \"When an insect cell is used as a host, a transfer vector is, for example, pVL1392, pVL1393 or pBlueBacIII (Invitrogen) and a virus for infection is, for example, a Vaculovirus which infects insects of Mamestra brassicoe family; Autographa california nuclear polyhedrosis virus (AcMNPV) Bac-N-Blue DNA.\",\n \"A transformation method of an insect cell is, for example, a method described in Baculovirus Expression Vector: A Laboratory Manual (1992) (W.H.\",\n \"Freeman and Company), Molecular Cloning: A Laboratory Manual, Second Edition (1989) (Cold Spring Harbor Laboratory Press), Current Protocols in Molecular Biology (1994) (Wiley-InterScience) or Biotechnology, 6, 47 (1988).\",\n \"A transfer vector including a target gene and baculovirus DNA for infection to an insect cell are added into a culture and a virus expressing a target gene produced by recombinant infects an insect cell to be expressed a polypeptide.\",\n \"An insect cell as a host is, for example, an established cell from Spodoptera frugiperda (Mamestra brassicoe) or an established cell from Trichoplusia ni.\",\n \"For example, a cell from S. frugiperda includes Sf9 (ATCC: CRL-1711, an ovary cell) or Sf21 (an ovary cell) and a cell strain from T.ni is, for example, High Five or BTI-TN-5B1-4 (an egg cell, Invitrogen).\",\n \"Any method to introduce into a host can be used as an introduction method of a recombinant vector, for example, a calcium phosphate method (JP1990-22705) or a lipofection method (Proceedings of the National Academy of Sciences USA, 84, 7413 (1987).).\",\n \"And an electroporation (Cytotechnology, 3, 133 (1990).)\",\n \"can be used as well as an animal cell.\",\n \"(vi) A Culture Method When a transformant having a recombinant vector with an inserted DNA encoding KIAA1036 polypeptide of the present invention is a cell such as Escherichia coli, yeast, an animal cell or a plant cell, a cultivation by a usual culture method suited to all kinds of hosts is held.\",\n \"And the polypeptides are produced, accumulated and collected from transformants or a culture solution to produce the polypeptide.\",\n \"When transformants are animals or plants, they are cultured by a usual growth method suited to all kinds of hosts.\",\n \"And the polypeptides are produced, accumulated and collected from animals or plants to produce the polypeptide.\",\n \"When a host is an animal, for example, a nonhuman transgenic animal having polynucleotides of the present invention is cultured.\",\n \"And KIAA1036 polypeptides encoded by the recombinant DNA are produced and accumulated in the animal and they are collected from an animal to produce KIAA1036 polypeptides.\",\n \"A production/accumulation place in an animal is, for example, milk, sputum or egg of the animal.\",\n \"When a host is a plant, for example, a transgenic plant having KIAA1036 polynucleotides of the present invention is cultured.\",\n \"And KIAA1036 polypeptides encoded by the recombinant DNA are produced or accumulated in a plant and collected from a plant to produce KIAA1036 polypeptides.\",\n \"When a host is a prokaryote such as Escherichia coli or a eucaryote such as yeast, for example, transformants having polynucleotides of the present invention are cultured in a medium.\",\n \"And KIAA1036 polypeptides encoded by the recombinant DNA are produced or accumulated in a culture solution and collected from the culture to produce polypeptides of the present invention.\",\n \"The method that transformants of KIAA1036 of the present invention are cultured in a medium is accomplished by a usual method for cultivation.\",\n \"When a host is prokaryote such as Escherichia coli or a eucaryote, a natural medium or a synthetic medium can be used as a medium that obtained transformants are cultured if it has a carbon source, a nitrogen source and mineral that a host can assimilate and it is a medium that cultivation of transformants is held efficiently.\",\n \"For example, YT medium including bactotryptone, yeast extract and sodium chloride is preferable as a medium when transformants that hosts are Escherichia coli are cultured.\",\n \"Any carbon source that each microorganism can assimilate can be used, for example, glucose, fructose, sucrose, syrup including them, carbohydrate such as starch or starch hydrolysate, an organic acid such as acetic acid or proprionic acid or alcohol such as ethanol or propanol.\",\n \"A nitrogen source is, for example, all kinds of inorganic acids such ammonia, ammonium chloride, ammonium sulfate, ammonium acetate, ammonium phosphate, ammonium salts of organic acids, other nitrogenous substances, peptone, meat extract, yeast extract, Corn Steep Liquor, casein hydrolysate, soybean cake, soybean cake hydrolysate, all kinds of fermentative bacteria or the digest.\",\n \"A mineral is, for example, potassium phosphate monobasic, potassium phosphate dibasic, magnesium phosphate, magnesium sulfate, sodium chloride, ferrous sulfate, manganese sulfate, copper sulfate or calcium carbonate.\",\n \"Cultivation is held under an aerobic condition such as a shaking culture or a submerged culture.\",\n \"An incubation temperature is preferably 15 to 40° C. and a culture time is usually 5 hours to 7 days.\",\n \"During cultivation, pH is kept from 3.0 to 9.0.Adjustment of pH is held with mineral or organic acid, alkali solution, urea, calcium carbonate or ammonia.\",\n \"And an antibiotic such as ampicillin or tetracycline can be optionally added into a medium during cultivation.\",\n \"When microorganisms transforming with an expression vector with an inductive promoter are cultured, inducers can be optionally added into a medium.\",\n \"For example, when transformants transforming with an expression vector with lac promoter are cultured, isopropyl β-D-thiogalactopyranoside can be added into a medium.\",\n \"When transformants transforming with an expression vector with trp promoter are cultured, indoleacrylic acid can be added into a medium.\",\n \"Cells and organs of plants introduced genes can do a mass culture with jar fermenter.\",\n \"A culture medium is, for example, popularly used Murashige & Skoog (MS) medium, White medium, or a medium that plant hormones such as oxine or cytokinin are added into these mediums.\",\n \"When transformants for production of KIAA1036 polypeptide are animal cells, a medium that cells are cultured is popularly used RPMI1640 medium (The Journal of the American Medical Association, 199, 519 (1967).\",\n \"), MEM medium (Science, 130, 432 (1959).\",\n \"), D-MEM medium (Virology, 8, 396 (1959).\",\n \"), 199 medium (Proceedings of the Society for the Biological Medicine, 73, 1 (1950).)\",\n \"or a medium that fetal calf serum (FCS) is added into these mediums.\",\n \"Cultivation held usually in pH 6 to 8, at 25 to 40° C., in the presence of 5% CO2 for 1 to 7 days.\",\n \"And an antibiotic such as kanamycin, penicillin or streptomycin can be optionally added into a medium during cultivation.\",\n \"When transformants are insect cells, a culture medium is popularly used TNM-FH medium (Pharmingen), Sf-900II SFM medium (Invitrogen), ExCell400, ExCell405 (JRH Biosciences Inc.) or Grace's Insect Medium (Nature, 195, 788 (1962).).\",\n \"(vii) The Method of Production Transformants are cultured and KIAA1036 polypeptides are isolated and purified from a culture to produce KIAA1036 polypeptides.\",\n \"A method of isolation/purification of KIAA1036 polypeptides can be usual method widely known in this field, for example, a method of isolation/purification of an enzyme or a method of purification of transglucosylase by Sandler (Methods in Enzymology, 83, 458).\",\n \"When KIAA1036 polypeptides are produced and accumulated as dissolved polypeptides, a culture solution that transformants are cultured as mentioned above is separated to cells or fungus body and a medium, for example, by a centrifugal separation.\",\n \"When KIAA1036 polypeptides exist in host cells, after cells or fungus bodies extracted are washed with an appropriate buffer such as STE solution and broken into pieces by ultrasonic waves, French press, Manton Gaulin homogenizer or Dynomill, KIAA polypeptides can be obtained as an acellular solution by a centrifugal separation or a filtration.\",\n \"The suitable quantity of surfactant can be included in a buffer for separation/purification of KIAA1036 polypeptide.\",\n \"For example, sodium lauryl sulfate (SDS) or Sodium N-Dodecanoylsalcosinate (salcosiyl) can be used.\",\n \"A method of separation/purification of target proteins included in an obtained crude material can be accomplished with the combination of all kinds of well-known methods of separation/purification.\",\n \"The well-known method is, for example, a solvent extraction method, a salting-out method with ammonium sulfate, a dialysis, an sedimentation with an organic solvent, an ultrafiltration method, a gel filtration, all kinds of chromatography such as a diethylaminoethyl (DEAE)-sepharose chromatography, an anion chromatography or an ion exchange chromatography using lysine such as DIAION HPA-75 (Mitsubishi Chemical Corporation), a cation chromatography using lysine such as S-Sepharose FF (Pharmacia), a hydrophobic chromatography or an affinity chromatography such as butylsepharose or all kinds of electrophoresis such as a SDS-polyacrylamide gel electrophoresis or an electro-focussing electrophoresis.\",\n \"An affinity chromatography can be accomplished by using antibodies against KIAA1036 polypeptide.\",\n \"When KIAA1036 polypeptides are produced and accumulated as insoluble polypeptides, cells or fungus bodies are separated as mentioned above and broken into pieces by an appropriate method.\",\n \"Then a division including the polypeptides is collected.\",\n \"A collected sample is solubilized with a solubilizer like a surfactant such as sodium lauryl sulfate (SDS) or Sodium N-Dodecanoylsalcosinate (salcosiyl).\",\n \"After the solubilized solution is diluted or dialyzed to the concentration that a solubilizer is not or almost not included and the polypeptide is constructed to a normal stereo structure, a purification sample can be obtained by a method of separation/purification as mentioned above.\",\n \"And KIAA1036 polypeptides can be produced as fusion proteins with the other proteins and purified by an affinity chromatography with substances having affinity with the fusion proteins (Yamakawa Akio, “Experimental Medicine”, 13, 469-474 (1995).).\",\n \"An addition protein used as a fusion protein is, for example, protein A or FLAG (Proceedings of the National Academy of Sciences USA, 86, 8227 (1989)., Genes Development, 4, 1288 (1990)., JP1993-336963 or JP1994-823021).\",\n \"When protein A are used, fusion proteins with KIAA1036 polypeptides and protein A can be produced and purified by an affinity chromatography with immunoglobulin G. When FLAG peptides are used, fusion proteins with KIAA1036 polypeptides and FLAG can be produced and purified by an affinity chromatography with anti-FLAG antibodies.\",\n \"KIAA1036 polypeptides can be produced as well as a well-known method with in vitro transcription/translation system (Journal of Biomolecular NMR, 6, 129-134 (1995)., Science, 242, 1162-1164 (1988), or The Journal of Biochemistry, 110, 166-168 (1991).).\",\n \"KIAA1036 polypeptides can be chemosynthesized on the basis of the amino acid sequence by a chemical synthesis method such as Fmo method (Fluorenylmethyl oxycarbonyl method) or tBoc method (t-butyl oxycarbonyl method), or by a peptide synthetic equipment on the market such as, for example, APEX396 (Advanced Chemtech), 433A (Applied Biosystems), PS3 (Protein Technologies), 9050 (Perseptive) or PSSM-8 (Shimazu corporation).\",\n \"A structural analysis of KIAA1036 polypeptide can be accomplished by a usual method in the field of protein chemistry, for example, a method described in “A protein structural analysis for gene cloning” (Hisashi Hirano, TOKYO KAGAKU DOZIN Co., LTD., 1993).\",\n \"The chordin activity of a polypeptide of the present invention can be measured as well as a well-known assay (The Journal of Biological Chemistry, 258, 9893-9898 (1983)., The Journal of Biological Chemistry, 262, 15649-15658 (1987)., The Journal of Biological Chemistry, 273, 58-65 (1998)., The Journal of Biological Chemistry, 273, 433-440 (1998)., The Journal of Biological Chemistry, 273, 12770-12778 (1998)., Archives of Biochemistry and Biophysics, 270, 630-646 (1989)., Archives of Biochemistry and Biophysics, 274, 14-25 (1989), or JP1994-181759).\",\n \"(3) A Method of Production of a Variant Polypeptide Mutation A deletion, a substitution or an addition of an amino acid of KIAA1036 polypeptide is accomplished by a widely known technique, a site-specific potentially mutagenic method.\",\n \"A deletion, a substitution or an addition of 1 or few amino acid(s) can be prepared as well as the method described in Molecular Cloning: A Laboratory Manual, Second Edition (1989) (Cold Spring Harbor Laboratory Press.\",\n \"), Current Protocols in Molecular Biology (1994) (Wiley-InterScience), Nucleic Acids Research, 10, 6487 (1982)., Proceedings of the National Academy of Sciences USA, 79, 6409 (1982)., Gene, 34, 315 (1985)., Nucleic Acids Research, 13, 4431 (1985)., Proceedings of the National Academy of Sciences USA, 82, 488(1985)., Proceedings of the National Academy of Sciences USA, 81, 5662 (1984)., Science, 224, 1431 (1984)., WO85/00817 or Nature, 316, 601 (1985).\",\n \"(4) A Production of an Antibody Against KIAA1036 Polypeptide (i) A Production of a Polyclonal Antibody Antibodies can be produced by giving all length of KIAA1036 polypeptides, a part of the peptides or polypeptides including a part of the peptides as antigens to a mammal.\",\n \"The peptide itself and a carrier, for example, a carrier combined with cattle serum albumin (BSA), keyhole limpet hemocyanin (KLH) or bovine thyroglobulin (BTG) can be used as an antigen.\",\n \"To enhance immune reactions with antigens, for example, a complete Freund adjuvants (CFA) and an incomplete Freund adjuvants (IFA) can be given.\",\n \"A mouse, a rat, a rabbit, a goat or a hamster can be used as a mammal to immunize.\",\n \"Polyclonal antibodies can be produced, for example, as well as the method by Lane et al.\",\n \"(Antibodies: A Laboratory Manual, Second Edition (1989)(Cold Spring Harber Laboratory Press)).\",\n \"After the first immunization, a mammal is immunized by an appropriate antigen 3 to 10 times at 1- to 2-week intervals.\",\n \"And then a serum from the mammal is extracted and purified to produce the antibodies.\",\n \"The antigens are given 3 to 10 times at 1- to 2-week intervals.\",\n \"A preferable dosage of the antigens is 50 to 100 μg at one time per an animal.\",\n \"When peptides are used, peptides covalently bonded to appropriate carriers are preferably used as antigens.\",\n \"Peptides as antigens can be synthesized by a method of genetic engineering or a peptide synthesizer.\",\n \"Three to seven days after immunization, the blood from venous plexus of eyegrounds is collected and the responsiveness of the serum against the antigens can be measured by an enzyme-liked-immunosorbent assay (Enzyme-liked-immunosorbent assay (ELISA): Igaku-Shoin Ltd. (1976), Antibodies: A Laboratory Manual, Second Edition (1989)(Cold Spring Harbor Laboratory Press).\",\n \"Blood is collected from a mammal immunized and an antibody titer is measured.\",\n \"Blood is collected when the immunized animal shows a sufficient antibody titer, and then polyclonal antibodies can be prepared from the serum.\",\n \"Separation and purification of polyclonal antibodies can be accomplished by an individual or a combination of all kinds of chromatography such as a centrifugal separation, salting-out with ammonium sulfate, precipitation with caplyric acid (Antibodies: A Laboratory Manual, Second Edition (1989)(Cold Spring Harbor Laboratory Press), DEAE-sepharose column, anion exchange column, protein A column or G-column or a gel filter column.\",\n \"(ii) A Production of a Monoclonal Antibody (a) A Preparation of an Antibody Producing Cell After a sufficient antibody titer in (i) is obtained, spleen or lymph node is extracted from the mammal.\",\n \"And then a monoclonal antibody producing hybridoma can be obtained by fusing an antibody-producing cell from the spleen or lymph node with a myeloma cell.\",\n \"As for the myeloma cell, cell strains established from a mouse or a rat can be used.\",\n \"Cell fusion can be done according to an already known method, for example, a method by Kohler and Milstein (Nature, 256, 495-497 (1975).).\",\n \"KIAA1036 polypeptides, a part of the peptides or polypeptides including the part of the peptides are immunized to a rat.\",\n \"Three to seven days after a rat showed a sufficient antibody titer, the rat is immunized with the antigen for the last time, and its spleen is extracted as antibody producing cells.\",\n \"The spleen is cut to pieces in MEM medium (Nissui Pharmaceutical Co. Ltd.) and untied with tweezers.\",\n \"And then a precipitant is obtained after a centrifugation at 1,200 rpm for 5 minutes.\",\n \"Splenocytes is separated by treating the precipitant with Tris-ammonium chloride buffer (pH 7.65) for 1 to 2 minutes to remove red blood cells.\",\n \"The splenocytes are washed with MEM medium 3 times and are used as antibody producing cells.\",\n \"(b) A Preparation of a Myeloma Cell An established cell line from mouse or rat is used as a myeloma cell.\",\n \"For example, it is a myeloma cell of 8-azaguanine resistance mouse (from BALB/c), P3-X63Ag8-U1 strain (described blow as P3-U1) (Current Topics Microbiological Immunology, 81, 1 (1978), or European Journal of Immunology, 6, 511 (1976).\",\n \"), SP2/0-Ag14 strain (described blow as SP-2) (Nature, 276, 269 (1978).\",\n \"), P3-X63-Ag8653 strain (described blow as 653) (Journal of Immunology, 123, 1548 (1979).)\",\n \"or P3-X63-Ag8 (described blow as X63) (Nature, 256, 495 (1975).).\",\n \"These cell strains are subcultured in a 8-azaguanine medium (a normal medium including 15 μg/ml 8-azaguanine (RPMI1640 medium including 1.5 mM glutamine, 5×10−5M 2-mercaptoethanol, 10 μg/ml gentamysin and 10% FCS made by CSL)) and cultured in a normal medium for 3 to 4 days before cell fusion.\",\n \"2×107 or more cells are prepared for cell fusion.\",\n \"(c) A Production of Hybridoma Antibody producing cells prepared in (a) and myeloma cells prepared in (b) are washed with MEM medium or PBS (per 1 L; 1.83 g sodium phosphate dibasic, 0.21 g monobasic potassium phosphate, 7.65 g NaCl, pH 7.2) and mixed as the number of antibody producing cells is 5 to 10 times larger than that of the myeloma cells.\",\n \"After a centrifugal separation at 1,200 rpm for 5 minutes, a precipitant is obtained.\",\n \"The precipitated cells are well untied and 0.2 to 1 ml of polyethylene glycol solution (2 g polyethylene glycol-1000 (PEG-1000), 2 ml MEM medium, 0.7 ml dimethyl sulfoxide (DMSO)) per 108 antibody producing cells is added to the cells with stirring at 37° C. And then 1 to 2 ml of MEM medium is added for several times every 1 to 2 minutes.\",\n \"The solution is prepared with MEM medium to 50 ml in total.\",\n \"After a centrifugal separation at 900 rpm for 5 minutes, a precipitant is obtained.\",\n \"100 ml of HAT medium (normal medium including 10−4 M hypoxanthine, 1.5×10−5 M thymidine and 4×10−7 M aminopterin) is added to a precipitant and the precipitant is untied slowly and suspended.\",\n \"The suspension is poured into the 96-well culture plate at 100 μl per well and cultured at 37° C. in the presence of 5% CO2 for 7 to 14 days.\",\n \"By the method described in “enzyme immunoassay” (Antibodies: A Laboratory Manual, Second Edition (1989)(Cold Spring Harbor Laboratory Press)), hybridomas producing antibodies specifically reacting with KIAA1036 polypeptides are selected.\",\n \"(5) an Immunoassay of KIAA 1036 Polypeptide The method to measure polypeptides or a part of the peptides by using antibodies that recognize the polypeptides or a part of the peptides and are directly or indirectly bonded to enzymes, fluorescent substances, radioisotopes or latexes can be used as an immunoassay of KIAA 1036 polypeptides.\",\n \"The assay is, for example, ELISA or a chemiluminescence method detecting enzyme activities such as horseradish peroxidase or alkaline phosphatase, FITC method detecting fluorescent tags such as luminol or GFP (Green Fluorescence Protein), RIA method detecting radioisotope tags such as 125I or a latex agglutination method detecting binding with latex.\",\n \"And the assay is, for example, Western blotting or an immune structure dyeing method.\",\n \"Furthermore, KIAA1036 polypeptides or a part of peptides can be quantitated by the assay.\",\n \"Antibodies for an immunoassay can be immobilized to a solid phase carrier and the trapped polypeptides can be detected by using secondary antibodies with a reporter group or using reagents.\",\n \"A competitive method that KIAA1036 polypeptides are labeled with a reporter group, reacted with antibodies and a sample and bonded with immobilized antibodies can be used to detect.\",\n \"The level of inhibition the binding between the labeled polypeptides and antibodies by KIAA1036 polypeptides of a sample is shown by reactivity with immobilized antibodies of a sample and the concentration of KIAA1036 polypeptides in a sample can be calculated.\",\n \"Any substance, to which antibodies can attach and which is widely known to a parson having ordinary skill in the art, can be used as a solid phase carrier.\",\n \"The substance includes, for example, a microtitre plate, a membrane such as a nitrocellulose membrane, bead, disk, glass, glass fiber, plastic material such as latex, polystyrene or polyvinyl chloride.\",\n \"Magnetic particles or fiber optical sensors (U.S. Pat.\",\n \"No.\",\n \"5,359,681) can be used.\",\n \"A well-known method used in this field can be used as a method which antibodies are immobilized to a carrier.\",\n \"In this description, “solid phase” means immobilization by a physical method such as adsorption or a chemical binding by a covalent bond between an antibody and a functional group on a carrier.\",\n \"An antibody and a functional group on a carrier can be bonded directly or through a cross-linking agent.\",\n \"Immobilization by a physical method can be accomplished by appropriately diluted antibodies contacted with a carrier, preferably, a microtiter plate or a membrane in an appropriate buffer for an appropriate time.\",\n \"The contact time varies depending on the temperature, but it is typically between about 1 hour and 1 day.\",\n \"About 10 ng to 1 μg, preferably, about 100 to 200 ng of antibodies is added and immobilized on each well of a microtiter plate made of plastic such as polystyrene or polyvinyl chloride.\",\n \"Immobilization by a chemical method can be accomplished by a reaction of a carrier and functional groups of antibodies, for example, a reaction of a carrier and a two-functional reagent that reacts with both hydroxyl groups and amino groups and a carrier.\",\n \"For example, antibodies can be immobilized to a carrier having an appropriate polymer coat with a covalent bond by using benzoquinone or a condensation between aldehyde groups on a carrier and an amine or an active hydrogen on a combination partner.\",\n \"The method can be accomplished by using for example, Pierce immunotechnology Catalog and Handbook (1991) A12 to A13 as a reference.\",\n \"A carrier-immobilized antibody is treated to inhibit physical adsorption of other polypeptides by a well-known method for a parson having ordinary skill in the art with an appropriate blocking reagent, for example, cattle serum albumin or Tween 20 (Sigma-Aldrich).\",\n \"A carrier-immobilized antibody is reacted with a sample and polypeptides of the present invention and antibodies are combined.\",\n \"A sample can be appropriately diluted with an appropriate diluent, for example, phosphate buffered saline solution (PBS).\",\n \"A reaction time of a sample and antibodies should be enough to detect the presence of polypeptides of the present invention in a sample obtained from an individual having a cancer, preferably, a time to achieve at least 95% of binding level compared to the level at which bound and not-bound polypeptides are equilibrated.\",\n \"A time to reach to equilibrium can be easily decided by measuring the binding level by the time.\",\n \"Substances other than bound polypeptides can be removed by washing a solid carrier with an appropriate buffer, for example, PBS (including 0.1% Tween 20).\",\n \"Labeled secondary antibodies are reacted with a solid carrier.\",\n \"The labels are preferably enzymes such as horseradish peroxidase, ground substances, supplemental elements, inhibitors, pigments, radioisotopes, coloring substances or fluorescent substances.\",\n \"The binding between antibodies and labels can be accomplished by a well-known method for a parson having ordinary skill in the art.\",\n \"The secondary antibodies are reacted for a sufficient time to bind to complexes, which include immobilized antibodies and polypeptides of the present invention.\",\n \"An appropriate time can be easily decided by measuring binding level by the time.\",\n \"The non-binding secondary antibodies can be removed by washing a solid carrier with an appropriate buffer, for example, PBS (including 0.1% Tween 20).\",\n \"The method of detection of labels of the secondary antibodies is different by a kind of labels.\",\n \"When radioisotopes are used as labels, detection by a scintillation counter or an autoradiography can be used.\",\n \"When pigments, coloring substances or fluorescent substances are used as labels, detection by a spectrophotometer can be used.\",\n \"When enzymes are used as labels, ground substances against the enzymes are added and reacted for a fixed time and the products are detected by a spectrophotometer.\",\n \"Labels and secondary antibodies can bind directly or indirectly by an avidin-biotin method.\",\n \"When they bind indirectly, one part of the avidin-biotin is bound to a secondary antibody and another is bound to a label.\",\n \"KIAA1036 polypeptides can be detected by a flow through test or a strip test.\",\n \"In a flow through test, a sample is added to a nitrocellulose membrane on which antibodies are immobilized, and when a sample passes through the membrane, polypeptides bind to the immobilized antibodies to form immune complexes.\",\n \"And when a solution including labeled secondary antibodies pass through the membrane, it binds to the immune complexes.\",\n \"In a strip test, once a sample is added, the sample pass through a region including labeled antibodies, and polypeptides bind to labeled antibodies to form immune complexes.\",\n \"When a sample pass through a region including a solid phase antibodies, polypeptides bind to the immune complexes.\",\n \"The quantity of secondary antibodies detected in the region with immobilized antibodies shows the presence or absence of cancer.\",\n \"Labels in the detection part can be visualized for confirmation.\",\n \"If it can not be confirmed, it shows a negative result.\",\n \"A quantity of antibodies immobilized to a membrane is preferably about 25 ng to 1 μg and more preferably about 50 ng to 1 μg.\",\n \"(6) An Assay of mRNA Using oligonucleotides prepared from KIAA1036 polynucleotides, mRNAs are quantitated by Northern hybridization or RT-PCR, and then expression level of a gene encoding KIAA1036 polypeptide can be quantitated.\",\n \"When mRNAs are quantitated with normal or disease model animals, for example, mice, rats, rabbits, sheep, pigs, cattle, cats, dogs or monkeys, an agent such as an anticancer agent is optionally given.\",\n \"After a fixed time, organs or tissues such as blood, brain, stomach or kidney is isolated and cells are prepared.\",\n \"mRNAs can be extracted from obtained cells by a widely known and usual method in this field and quantitated/analyzed by RT-PCR or Northern blot hybridization.\",\n \"When mRNAs from transformants, which express KIAA1036 polypeptides or a part of the peptides, are quantitated, mRNAs can be extracted from the transformants by a widely known and usual method in this field and quantitated/analyzed by RT-PCR or Northern blot hybridization.\",\n \"(7) Identification of a Structure of the Gene Recently, a lot of sequences of human genome whose functions are unknown are registered on databases.\",\n \"Therefore, by comparing KIAA1036 polynucleotide sequence and a sequence of human genome registered on databases, human gene encoding KIAA1036 polypeptide can be identified and a structure of the gene can be clarified.\",\n \"If a genomic sequence, which coincides with cDNA sequence, is registered, a promoter region and a structure of exon and intron of the gene encoding a polypeptide of the present invention can be screened by comparing cDNA sequence and a sequence of the gene.\",\n \"By using KIAA1036 polynucleotide as a probe and a widely known and usual method in this field (Division of Oncology, Institute of Medical Science, Univ.\",\n \"of Tokyo, New Protocol for Molecular and Cellular Biology, Shujunsha (1993).\",\n \"), a promoter region of the gene can be obtained.\",\n \"Promoter regions are all of the promoter regions concerned with transcription of a gene encoding KIAA1036 polypeptide in mammal cells.\",\n \"(8) A Method of Detection of Disease KIAA1036 polypeptides can be used for diagnosis of diseases relating to neovaucularization, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer because the expression is increased in angiogenesis models.\",\n \"Furthermore, because the expression of the polypeptides is increased in ovarian cancer and large bowel cancer tissue, it can be used for diagnosis of these cancers.\",\n \"Polypeptides of the present invention or polynucleotides such as mRNAs are specifically detected from biological samples, for example, cancer or normal tissue specimen, dissected tissue, blood, lymph node, serum, urine or other tissues obtained from a patient and their homogenates or extracts.\",\n \"Then presence or absence of cancer in a patient can be decided by comparing the expression level in a patient with a level in a normal person or a normal tissue.\",\n \"The method of detection of disease includes, for example, an immunoassay or a molecular-biological method of detection of the polypeptides.\",\n \"An immunoassay of the polypeptides includes, for example, sandwich ELISA.\",\n \"In this method, a pair of antibodies recognizing different epitopes of the polypeptide are used and one antibody is labeled directly or indirectly with an enzyme.\",\n \"Another method is a competitive RIA by using the polypeptides labeled with radioisotopes such as 125I and antibodies recognized them.\",\n \"A molecular-biological method of detection includes, for example, in situ hybridization or PCR.\",\n \"PCR can be used to amplify cDNA prepared from mRNA isolated from a biological sample from a patient.\",\n \"A primer for PCR can be chemically synthesized on the basis of DNA sequence of SEQ ID NO: 1.Preferred is Primer1 which is an oligonucleotide sequence of SEQ ID NO: 4,5′-GTTCAGGACTGTCTTTCAGC-3′, or Primer2 which is an oligonucleotide sequence of SEQ ID NO: 5,5′-GTCAATACTGATGGACTTGC-3′.\",\n \"cDNA amplified by PCR can be separated by gel electrophoresis and then turned visible by the method widely known to a parson having ordinary skill in the art.\",\n \"Usually, samples are appropriately prepared from a pair of tissues, which are a cancer tissue and a normal tissue from the same individual or different individuals and subjected to PCR.\",\n \"An amplification reaction by PCR is preferably held with a serial two orders dilution of template cDNAs.\",\n \"In some dilution points, more than two-fold increase in expression of a cancer sample compared to a normal sample is considered as a positive result.\",\n \"The presence or absence of cancer is judged by measuring markers of the present invention detected in a normal sample and a patient sample and comparing.\",\n \"Generally, a mean value of markers from a healthy person without cancer is considered as a cut-off value and the cut-off value is compared to detected value of markers from a patient sample.\",\n \"Generally, when the detected value is over three times of the standard deviation plus a cut-off value, the sample is considered as a positive about cancer.\",\n \"And a cut-off value is decided by Receiver Oparator Curve described in Sackett et al., Clinical Epidemiology: A Basic Science for Clinical Medicine 106-107, (1985)(Little Brown and Co).\",\n \"(9) A Diagnostic Kit KIAA1036 polynucleotides can be used for a diagnosis of neovascularization related disease, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer such as ovarian cancer or large bowel by Northern hybridization or PCR.\",\n \"And a diagnosis of neovascularization related disease, for example, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer such as ovarian cancer or large bowel can be held by an immunologic method with antibodies against KIAA1036 polypeptides.\",\n \"Therefore, in the case of a diagnosis with polynucleotides, labeled KIAA1036 polynucleotides are contained in a kit.\",\n \"And in the case of a diagnosis with antibodies, KIAA1036 polypeptides are contained as standard antigens in addition to antibodies against KIAA1036 polypeptides.\",\n \"In a kit, a standard curve can be contained.\",\n \"(10) The Method to Inhibit mRNA Translation Giving KIAA1036 polynucleotides can inhibit the production of KIAA1036 polypeptides.\",\n \"Namely, by using KIAA1036 polynucleotides, transcription of DNA encoding KIAA1036 polypeptide and translation of mRNA encoding the polypeptide can be inhibited.\",\n \"(11) The Screening Method of Binding Substances As a reagent for search or screening binding substances to markers of the present invention, KIAA1036 polypeptides are useful.\",\n \"Namely, the screening method of binding substances to markers of the present invention, which comprises contact between markers and subjects, is provided.\",\n \"Subjects include, for example, chemical substances synthesized naturally or chemically, proteins, tissue extracts of a mammal such as human, mouse, rat, pig, cattle, sheep or monkey or cell culture.\",\n \"These subjects are added to markers of the present invention and the mixture is separated with measuring DNA synthesis activity of vascular endothelial cells.\",\n \"Finally, single ligands can be obtained.\",\n \"For example, the screening method of binding substances of the present invention includes a method by using KIAA1036 polypeptides or the partial peptides, by constructing an expression system of recombinant polypeptides and using a binding assay with the expression system, by binding to KIAA1036 polypeptides and measuring cell stimulatory activity, for example, activity to promote or inhibit DNA synthesis by bringing BrdU (bromodeoxyuridine) or cell migrating activity or by measuring visually network formation of vascular endothelial cells with microscope.\",\n \"At first, any polypeptide comprising polypeptide B can be used as polypeptides for the screening method of the binding substances but polypeptides expressed massively with animal cells is preferable.\",\n \"To produce polypeptide B, the above mentioned expression method is used, but the method is preferable when it includes expressing DNA encoding the polypeptide in mammal cells or insect cells.\",\n \"Generally cDNA is used as a fragment which encodes a protein to be expressed, but it should not be always cDNAs but fragment from genomic DNA or synthetic DNA can be used.\",\n \"To introduce DNA fragments encoding KIAA1036 polypeptides into host animal cells and effectively express them, it is preferable that the DNA fragments are inserted downstream of a polyhedrin promoter of nuclear polyhedrosis virus (NPV) of baculovirus which use insects as a host, a promoter from SV40, a promoter of retrovirus, a metallothionein promoter, a human heat-shock promoter, a cytomegalovirus promoter or a SR α promoter.\",\n \"The examination of quantity and quality of expressed polypeptides can be done by a well-known method.\",\n \"For example, it is the method described in The Journal of Biological Chemistry, 267, 19555 to 19559 (1992).\",\n \"Therefore, in a screening method of binding substances of the present invention, polypeptides or the partial peptides purified by the well-known method, cells including the polypeptides or their supernatants can be used as a thing including KIAA1036 polypeptides or the partial peptides.\",\n \"In a screening method of binding substances of the present invention, when cells including KIAA1036 polypeptides are used, that cells can be immobilized to agarose gel.\",\n \"The method of immobilization can be held by the well-known method.\",\n \"Cells including KIAA1036 polypeptides mean host cells with expressing KIAA1036 polypeptides.\",\n \"For example, Escherichia coli, Bacillus subtilis, yeast, insect cells and animal cells are used as the host cells.\",\n \"For the screening of binding substances to KIAA1036 polypeptides, appropriate polypeptide fractions and labeled test substances are needed.\",\n \"As a polypeptide fraction, for example, a natural polypeptide fraction or a recombinant polypeptide fraction having the same activity as a natural one is preferable.\",\n \"The same activity means, for example, the same ligand binding activity or the same signal transduction activity.\",\n \"The labeled test substances are, for example, substances labeled with 3H, 125I, 14C or 35S.\",\n \"For example, for a screening method of binding substances to KIAA1036 polypeptides, at first, a polypeptide sample is prepared by suspending cells including KIAA1036 polypeptides or a membrane fraction of the cells in a buffer which is suitable for a screening method.\",\n \"Any buffer, which does not inhibit combining a ligand and a polypeptide, for example, a phosphoric acid buffer that is pH 4 to 10, preferably pH 6 to 8 or a Tris-HCl buffer is used as a buffer.\",\n \"And to decrease non-specific binding, a surfactant, for example, CHAPS, Tween-80 (Kao-Atlas), digitonin or deoxycolate or all kinds of protein, for example, cattle serum albumin or gelatin can be added into the buffer.\",\n \"Furthermore, to repress degradation of receptors and ligands by protease, a protease inhibitor, for example, PMSF, leupeptin, E-64 (PEPTIDE INSTITUTE, INC.) or pepstatin can be added.\",\n \"Test substances are labeled with, for example, constant quantity, preferably 5000 cpm to 500000 cpm of 3H, 125I, 14C or 35S are coexistence in 0.01 to 10 ml of the polypeptide solution.\",\n \"To measure the quantity of non-specific binding (NSB), a reaction tube with excessive amount of unlabeled test substances is also prepared.\",\n \"The reaction is held at 0° C. to 50° C., preferably at 4° C. to 37° C., for 20 minutes to 24 hours, preferably for 30 minutes to 3 hours.\",\n \"After the reaction, the solution is filtered by glass fiber filter papers and washed with a suitable quantity of the same buffer.\",\n \"The radioactivity remaining on the glass fiber filter paper is measured by a liquid scintillation counter or a γ-counter.\",\n \"Test substances that the count (B-NSB), the result obtained by subtracting quantity of non-specific binding (NSB) from all quantity of binding (B), is over 0 cpm can be selected as binding substances to markers of the present invention.\",\n \"For the screening of binding substances to markers of the present invention, cell stimulatory activity through the polypeptides, for example, activity to promote or inhibit DNA synthesis by bringing BrdU (bromodeoxyuridine) or cell migrating activity can be measured by well-known method or a measuring kit on the market.\",\n \"And the activity can be measured by measuring visually network formation of vascular endothelial cells with microscope.\",\n \"For example, at first, cells including polypeptides are cultured on the multiwell plate.\",\n \"When the screening of binding substances is held, in advance, medium or buffer changes to fresh medium or appropriate buffer, which is non-toxic to cells and test substances are added.\",\n \"After incubation for a fixed time, cells are extracted or their supernatants are collected and the products are quantitated by the method for them.\",\n \"(12) A Kit for Screening of Binding Substances A kit for screening of binding substances bonded to markers of the present invention is a kit comprising cells expressing polypeptide B or the polypeptides or their supernatant fraction including the polypeptides.\",\n \"The examples of a kit for screening of binding substances are the followings.\",\n \"(i) A Reagent for Screening of Binding Substances As a screening solution and washing solution, a solution that Hank's Balanced Salt Solution (Invitrogen) added 0.05% cattle serum albumin (SIGMA-Aldrich) is sterilized by filtration with 45 μm filter, and it is kept at 4° C. or prepared just before use.\",\n \"(a) COS-7 Cells Expressing Polypeptides of the Present Invention are Cultured on a 12-well Plate with 5×105 Cells/well and Cultured at 37° C. for 2 Days.\",\n \"(b) A Labeled Subject A solution containing commercially available subjects labeled with 3H, 125I, 14C or 35S is kept at 4° C. or 20° C., and diluted to 1 μM with assay buffer just before use.\",\n \"Subjects with the low solubility to water are dissolved in dimethyl formamide, DMSO or methyl alcohol.\",\n \"(c) A Non-Labeled Subject The same things as labeled substances are prepared at 100 to 1000 times higher concentration.\",\n \"(ii) Assay (a) COS-7 cells expressing polypeptides of the present invention cultured on a 12-well plate for tissue culture are washed twice with assay buffer (iml) and then assay buffer is added (490 μl per well).\",\n \"(b) Labeled subjects (5 μl) are added and the solution is reacted at a room temperature for 1 hour.\",\n \"Non-labeled subjects (5 μl) are added to measure the quantity of unspecific binding.\",\n \"(c) The reaction solution is removed and the remaining cells are washed with a washing solution (1 ml) 3 times.\",\n \"Labeled subjects bound to cells are dissolved with 0.2M NaOH (included 1% SDS) and the solution is mixed with liquid scintillator A (4 ml) (Wako Pure Chemical Industries, Ltd.).\",\n \"(d) The radioactivity is measured by liquid scintillation counter (Beckman Coulter).\",\n \"(13) A Screening Method of Binding Activity Regulatory Substances A marker of the present invention is useful as a reagent to search or screen binding activity regulatory substances to the markers.\",\n \"As a screening method of binding activity regulatory substances of the markers and binding substances, namely, substances changing the binding activity, a recombinant expression system expressing markers of the present invention is constructed and bioassay system or binding assay system is used.\",\n \"Then substances changing the binding activity between binding substances and markers of the present invention, for example, peptides, proteins, nonpeptide substances, synthetic substances or ferment products can be efficiently screened.\",\n \"The substances comprise a substance which can enhance or reduce (a) cell stimulatory activity between binding substances and a marker of the present invention, for example, an activity to promote or inhibit DNA synthesis monitored by BrdU (bromodeoxyuridine) incorporation or an activity to promote or inhibit of cell migrating activity, or (b) the binding activity between a binding substance and a marker of the present invention.\",\n \"Namely, the present invention provides with a screening method of binding activity regulatory substances between binding substances and markers of the present invention characterized by comparing the quantity of binding of binding substances when (i) markers of the present invention and binding substances are contacted and (ii) markers of the present invention, binding substances and subjects are contacted.\",\n \"A screening method of binding activity regulatory substances of the present invention is characteristic of comparing the quantity of binding between the markers and binding substances by measuring, for example, cell stimulatory activity in the case of (i) and (ii).\",\n \"A screening method of binding activity regulatory substances of the present invention is, for example, (a) a method to screen binding activity regulatory substances which can change the binding activity between binding substances and KIAA1036 polypeptides, comprising measuring and comparing the quantity of binding of labeled substances to the polypeptides when labeled binding substances are contacted to KIAA1036 polypeptides and when labeled binding substances and subjects are contacted, (b) a method to screen binding activity regulatory substances which can change the binding activity between binding substances and polypeptides of the present invention comprising measuring and comparing the quantity of binding of label binding substances to the polypeptides in the cells or cell culture when cells expressing KIAA1036 polypeptides or the cell culture solution and labeled binding substances are contacted and when labeled binding substances and subjects are contacted or (c) a method to screen binding activity regulatory substances which can change the binding activity between binding substances and KIAA1036 polypeptides comprising measuring and comparing the quantity of binding of label binding substances to the polypeptides when polypeptides secreted in cell culture solutions by culturing transformants having KIAA1036 polynucleotides and label binding substances are contacted and when label binding substances and subjects are contacted.\",\n \"(14) A Kit for Screening of Binding Activity Regulatory Substances A kit for screening of binding activity regulatory substances changing the binding activity between KIAA1036 polypeptides and binding substances is a kit comprising cells including the polypeptides or a cell culture solution.\",\n \"The examples of a kit for screening of the present invention are the followings.\",\n \"(i) A Reagent for Screening of Binding Activity Regulatory Substances As a screening solution and washing solution, a solution that Hank's Balanced Salt Solution (Invitrogen) added 0.05% cattle serum albumin (SIGMA-Aldrich) is sterilized by filtration with 45 μm filter, and it is kept at 4° C. or prepared just before use.\",\n \"(a) KIAA1036 Polypeptide COS-7 cells expressing KIAA1036 polypeptides are cultured on a 12 well plate with 5×105 cells/well and cultured at 37° C. for 2 days.\",\n \"(b) A Labeled Subject A solution containing commercially available subjects labeled with 3H, 125I, 14C or 35S is kept at 4° C. or 20° C., and diluted to 1 μM with assay buffer and prepared just before use.\",\n \"Subjects with the low solubility to water are dissolved in dimethyl formamide, DMSO or methyl alcohol.\",\n \"(c) A Non-Labeled Subject The same things with labeled substances are prepared at 100 to 1000 times higher concentration.\",\n \"(ii) An Assay (a) COS-7 cells expressing polypeptides of the present invention cultured on a 12-well plate for tissue culture are washed twice with assay buffer (1 ml) and then assay buffer is added (490 μl per well).\",\n \"(b) Labeled subjects (5 μl) are added and the solution is reacted at room temperature for 1 hour.\",\n \"Non-labeled subjects (5 μl) are added to measure the quantity of non-specific binding.\",\n \"(c) The reaction solution is removed and the remaining cells are washed with a washing solution (1 ml) 3 times.\",\n \"Labeled subjects binding to cells are dissolved with 0.2M NaOH (included 1% SDS) and the solution is mixed with liquid scintillator A (4 ml) (Wako Pure Chemical Industries, Ltd.).\",\n \"(d) The radioactivity is measured by liquid scintillation counter (Beckman Coulter).\",\n \"(15) A Screening Method of Expression Regulatory Substances Polynucleotide A, polypeptide B or antibodies against the polypeptides are useful as a reagent to search or screen expression regulatory substances to markers of the present invention.\",\n \"Namely, a screening method of expression regulatory substances of the present invention is, for example, a screening method of expression regulatory substances of markers of the present invention by measuring quantity of mRNAs or proteins of KIAA1036 polypeptides or the partial peptides included in (i) blood of a non-human mammal, a specific organ, a tissue or cells isolated from the organ or (ii) transformants and so on.\",\n \"(16) A Kit for Screening of Expression Regulatory Substances A kit for screening of expression regulatory substances of KIAA1036 polypeptides or its salt is a kit comprising cells expressing polypeptide A or polypeptide B or cell culture solutions including the polypeptides.\",\n \"The Example of a kit for screening of the present invention is the following.\",\n \"(17) A Kit for Screening on the Basis of the Immunologic Assay An assay of KIAA1036 polypeptides is, for example, sandwich ELISA using two kinds of monoclonal antibodies that their epitopes are different in antibodies reacting with the polypeptide in the liquid phase, or a radioimmunoassay using KIAA1036 polypeptides labeled with a radioisotope such as 126I and antibodies specifically recognizing it.\",\n \"Therefore, in a case of diagnosis with antibodies, KIAA1036 polypeptide is included as standard antigens in addition to antibodies of the present invention.\",\n \"Furthermore, standard curve can be included in a kit.\",\n \"(18) A Kit for Screening on the Basis of a Molecular-Biological Assay The expression level of DNA encoding KIAA1036 polypeptide can be quantitated in mRNA level by Northern hybridization or PCR using oligonucleotides prepared from KIAA1036 polynucleotides.\",\n \"For example, (i) an agent, for example, an anticancer agent is given into a normal or a disease model non-human mammal (e.g., mouse, rat, rabbit, sheep, pig, cattle, cat, dog or monkey) and after a fixed time, blood, a specific organ, for example, brain, kidney, colon, ovary, tissues or cells isolated from an organ are obtained.\",\n \"mRNAs of a polypeptide of the present invention or the partial peptide included in the obtained cells can be for example, extracted by the extraction method widely known in this field, quantitated by the method such as TaqManPCR or analyzed by Northern blot widely known, or (ii) transformants expressing KIAA1036 polypeptides or the partial peptides are produced by the above mentioned method and mRNAs of KIAA1036 polypeptides or the partial peptides included in the transformant can be quantitated/analyzed.\",\n \"And in a case of diagnosis with polynucleotides, labeled KIAA1036 polynucleotides are included in a kit.\",\n \"(19) A Pharmaceutical Composition A pharmaceutical composition comprising at least one selected from substances such as polynucleotide A, a complementary polynucleotide to the polynucleotide, polypeptide B, its antibody or a gene therapeutic vector having polynucleotide A can be given independently as a therapeutic agent.\",\n \"But preferred is usually that a pharmaceutical composition is mixed with one or more pharmaceutically acceptable carrier(s) and provided as a pharmaceutical preparation produced by the any method widely known in the technical field of galenical pharmacy.\",\n \"Because KIAA1036 inhibit DNA synthesis in vascular endothelial cells and cell migrating activity, DNA synthesis inhibitors and cell migrating activity inhibitors are included.\",\n \"And substances obtained by a screening method or a kit for screening of the present invention and their salts are included.\",\n \"The substance is, for example, (a) a substance enhancing or reducing cell stimulation activity through binding between binding substances and KIAA1036 polypeptides, for example, DNA synthesis activity, migrating activity, network formation activity of vascular endothelial cells, (b) a substance enhancing or reducing a binding activity between a binding substance and KIAA1036 polypeptide or (c) a substance enhancing or reducing an expression level of KIAA1036 polypeptide.\",\n \"The substance is, for example, a low-molecular weight compound, a peptide, a protein, a nonpeptide substance, a synthetic substance or a ferment product.\",\n \"These substances can be new or well-known substances and natural or synthetic substances.\",\n \"Because agonists against KIAA1036 polypeptides have the same activity as bioactivation of binding substances against KIAA1036 polypeptides, they are useful as safe and low-toxic drugs for the binding substance activity.\",\n \"Because antagonists against KIAA1036 polypeptides can inhibit bioactivation of binding substances against KIAA1036 polypeptides, they are useful as safe and low-toxic drugs, which inhibit the binding substance activity.\",\n \"Substances enhancing binding activity between binding substances and KIAA1036 polypeptides are useful as safe and low-toxic drugs to enhance bioactivation of binding substances against KIAA1036 polypeptides.\",\n \"Substances reducing binding activity between binding substances and KIAA1036 polypeptides are useful as safe and low-toxic drugs to reduce bioactivation of binding substances against KIAA1036 polypeptides.\",\n \"And gene therapeutic expression vectors expressing KIAA1036 are included.\",\n \"Because KIAA1036 can be a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, it is suggested that KIAA1036 relates to those diseases and they are useful for gene therapies of these diseases.\",\n \"The gene therapeutic expression vectors are expression vectors having a part or all of a polynucleotide of KIAA1036 and when they are introduced into cells or tissues, they can normalize disorders which cause some natural or acquired disease at a gene level.\",\n \"In other words, the vectors can compensate for cells with normal genes, repair/modify defects of genes and regulate expression of genes.\",\n \"(20) A Therapeutic Agent A pharmaceutical composition comprising polypeptide B, an antibody against the polypeptide, polynucleotide A, complementary polynucleotide to the polynucleotide, a binding substance, a bond regulatory substance, an expression regulatory substance, a DNA synthesis inhibitor, a cell migrating activity inhibitor or a gene therapeutic vector can be given independently as a therapeutic agent.\",\n \"But preferred is usually that a pharmaceutical composition is mixed with one or more pharmaceutically acceptable carrier(s) and provided as a pharmaceutical preparation produced by the any method widely known in the technique field of galenical pharmacy.\",\n \"The most effective route of administration in treats is desirable as a route of administration, for example, an oral administration or a non-oral administration such as intraoral, tracheobronchial, intrarectal, subcutaneous, intramuscular or intravenous.\",\n \"A form of administration is, for example, spray, capsule, tablet, granule, syrup, emulsion, suppository, injection, ointment or tapes.\",\n \"An appropriate pharmaceutical preparation for an oral administration is, for example, emulsion, syrup, capsule, tablet, powder or granule.\",\n \"For example, a liquid preparation such as emulsion or syrup can be produced by using water, saccharide such as sucrose, sorbitol or fructose, glycol such as polyethylene glycol or propylene glycol, oil such as sesame oil, olive oil or soybean oil, antiseptic such as p-hydroxy ester benzoate or flavor such as strawberry or peppermint as a excipient.\",\n \"Capsule, tablet, powder or granule can be produced by using a vehicle such as lactose, dextrose, sucrose or mannitol, a disintegrating agent such as starch or sodium alginate, a lubricant such as magnesium stearate or talc, a binding agent such as polyvinyl alcohol, hydroxypropylcellulose or gelatin, a surface active agent such as fatty acid ester or a plasticizer such as glycerol as a excipient.\",\n \"An appropriate pharmaceutical preparation for non-oral administration is, for example, injection, suppository or spray.\",\n \"For example, an injection is prepared by using a carrier comprising salt solution, dextrose solution or their mixture.\",\n \"A suppository is prepared by using a carrier such as cacao butter, fat hydride or carboxylic acid.\",\n \"And a spray is prepared by using the substance or a carrier, which does not stimulate oral cavity and airway mucous membrane of a recipient and spread the substance as microparticles to become easy to absorb.\",\n \"The carrier includes, for example, lactose or glycerol.\",\n \"A pharmaceutical preparation such as aerosol or dry powder is possible if a nature of the substance and that of the used carrier are appropriate for it.\",\n \"And in these non-oral agents, a component used as an excipient in oral agents can be added.\",\n \"A substance obtained by using a screening method or a kit for screening of the present invention or the salt is a binding substance, a binding activity regulatory substance, an expression regulatory substance, a DNA synthesis inhibitor or a cell migrating activity inhibitor, for example, (a) a substance enhancing or reducing cell stimulation activity through bonds between binding substances and polypeptide B, for example, an activity to promote or inhibit DNA synthesis, which can be monitored by BrdU (bromodeoxyuridine) uptake, or an activity to promote or inhibit of cell migrating activity, (b) a substance enhancing or reducing a binding activity between binding substances and polypeptide B or (c) a substance enhancing or reducing an expression level of a marker of the present invention.\",\n \"The substance is a low-molecular weight compound, peptide, protein, non-peptide substance, a synthetic substance or a ferment product.\",\n \"These substances can be new or well-known substances and natural or synthetic substances.\",\n \"Because agonists against KIAA1036 polypeptides have the same activity as bioactivation of binding substances against KIAA1036 polypeptides, they are useful as safe and low-toxic drugs for the binding substance activity.\",\n \"Because antagonists against KIAA1036 polypeptides can inhibit bioactivation of binding substances against KIAA1036 polypeptide or something like this, they are useful as safe and low-toxic drugs, which inhibit the binding substance activity.\",\n \"Substances enhancing binding activity between binding substances and KIAA1036 polypeptides are useful as safe and low-toxic drugs to enhance bioactivation of binding substances against KIAA1036 polypeptides.\",\n \"Substances reducing binding activity between binding substances and KIAA1036 polypeptides are useful as safe and low-toxic drugs to reduce bioactivation of binding substances against KIAA1036 polypeptides.\",\n \"Substances reducing binding activity between binding substances and KIAA1036 polypeptides are useful as safe and low-toxic drugs to reduce bioactivation of binding substances against KIAA1036 polypeptides.\",\n \"And therapeutic agent KIAA1036 polypeptides for any kinds of diseases comprising substances changing the expression level of KIAA1036 polypeptides seem to have any important role, for example, a central nervous system as described above.\",\n \"Therefore an expression regulatory substance changing the expression level of KIAA1036 polypeptides or the partial peptide can be used as a therapeutic agent for a disease related insufficiency of KIAA1036 polypeptides.\",\n \"When the substance is used as a therapeutic agent for a disease relating insufficiency of KIAA1036 polypeptide, it can be turned a pharmaceutical preparation by the stereotyped method.\",\n \"For example, the substance can be used orally as a tablet with sugar-coating, a capsule, an elixir or a microcapsule as occasion demandsor parentally as a sterile solution with water or the other pharmaceutically acceptable solution or an injection of suspension.\",\n \"For example, the substances can be produced by mixing as the unitary dose form required to accomplish popularly acceptable pharmaceutical preparation with physiologically acceptable well-known carrier, spice, vehicle, antiseptic, stabilizer or binding agent.\",\n \"The amount of active principle in these pharmaceutical preparations is decided to be able to obtain an appropriate capacity in a directed range.\",\n \"An excipient, which can mix with tablet or capsule, is, for example, a binding agent such as gelatin, corn starch, tragacanth or Arabian gum, a vehicle such as crystal cellulose, a swelling agent such as corn starch, gelatin or alginic acid, a lubricant agent such as magnesium stearate, a sweetening agent such as sucrose, lactose or saccharin or a spice such as peppermint or cherry.\",\n \"In the case where the dosage form is a capsule, the above type of ingredients and a liquid carrier such as fat and oil can be included.\",\n \"An aseptic composition for the injection can be prescribed by the usual pharmaceutical preparation, for example, by dissolving or suspending an activity substance in vehicle such as water for the injection or natural plant oil such as sesame oil or coconut oil.\",\n \"An aqueous solution for the injection is, for example, an isotonic solution such as D-sorbitol, D-mannitol or sodium chloride including physiological salt solution, dextrose or adjuvant.\",\n \"It can be used with appropriate solubilizing agent, for example, alcohol such as ethanol, propylene glycol or polyethylene glycol, a nonionic surface-active agent such as polysorbate 80 or HCO-50.Oil solution is, for example, sesame oil or soybean oil and it can be used with a solubilizing agent such as benzyl benzoate or benzyl alcohol.\",\n \"And the above preventive/therapeutic agent can be mixed with, for example, a buffer such as phosphate buffer or sodium acetate buffer, a soothing agent such as benzalkonium chloride or procaine hydrochloride, a stabilizer such as human serum albumin or polyethylene glycol, a preservatives such as benzyl alcohol or phenol or an antioxidant.\",\n \"A prepared injection solution is usually filled up into an appropriate ampoule.\",\n \"(21) A Method of Treating Because the above-mentioned pharmaceutical composition can be a therapeutic agent for vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer, a method of treating disease such as vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or cancer is provided by giving that pharmaceutical composition by an appropriate method.\",\n \"It is desirable that the above-mentioned pharmaceutical composition is used in the most effectively medication method in treats.\",\n \"And the method is, for example, an oral administration or a non-oral administration such as intraoral, tracheobronchial, intrarectal, subcutaneous, intramuscular or intravenous.\",\n \"A therapeutic agent as described above is safe and low-toxic and can be given to, for example, a mammal such as human, rat, mouse, rabbit, sheep, pig, cattle, cat, dog or monkey.\",\n \"A dosage of a preventive or therapeutic agent per 1 time varies depending on medication subject, an object organ, a condition of a disease or a medication method.\",\n \"But in the case of a oral administration, for example, for a patient with hypertonia whose body weight is 60 kg, usually approximately 0.1 to 100 mg per a day, preferably approximately 1.0 to 50 mg, more preferably approximately 1.0 to 20 mg are given.\",\n \"For example, in the case of an injection of parentally giving, for example, for a patient with hypertonia whose body weight is 60 kg, usually approximately 0.01 to 30 mg per a day, preferably approximately 0.1 to 20 mg, more preferably approximately 0.1 to 10 mg are given by an intravenous injection.\",\n \"In the case of the other animal, the amount converted into the amount per kg of body weight can be given.\",\n \"The dosage or times of a dosage varies depending on treating effect targeted, medication method, treating period, age or body weight, but usually it is 10 μg to 8 mg/kg per a day for an adult.\",\n \"EXAMPLES The following examples are provided to exemplify and do not restrict the present invention.\",\n \"As a genetic engineering method, if there is not a special provision, the method described in Molecular Cloning: A Laboratory Manual, 2nd Edition (Cold Spring Harbor Laboratory), is used.\",\n \"Example 1 An Identification of Gene Whose Expression in Human Umbilical Vein Endothelial Cells (HUVEC) is Enhanced by the Stimulation of Vascular Endothelial Cell Propagation Factors (VEGF) Three Endocell HUVEC (KURABO) which is a human umbilical vein vascular endothelial cell were thawed and then inoculated on 16 culture plates coating with collagen (IWAKI) poured EBM medium including 10% fetal calf serum (FCS) (Sanko Junyaku).\",\n \"After culturing at 37° C. for 3 days, the medium was changed with the same medium and cultured for 2 days.\",\n \"After the cells reached to confluence, the medium was changed with M199 medium including 1% FCS (Nissui) and cells were cultured for 24 hours.\",\n \"For a culture with VEGF treatment, the medium was changed to M199 medium including 1% FCS and 1 nM human VEGF165 (R&D Systems).\",\n \"For a culture without VEGF (a control), medium was changed with the same medium not including human VEGF165.Cells were cultured at 37° C. and collected after 0 hour, 0.5 hours, 2 hours, 6 hours, 12 hours and 24 hours.\",\n \"Collected cells were washed with PBS twice and total RNA were prepared by ISOGEN (Nippon Gene) and mRNAs were prepared by oligo dT cellulose column.\",\n \"These mRNAs were analyzed by LifeArrayhumanUniGENE (Incyte Genomics) through Gene Expression Microarray analysis on assignment of Incyte Genomics.\",\n \"As a result, for 6393 human genes, gene expression level was calculated as a ratio of a expression level in VEGF-treated cells compared to one in untreated cells in each time point.\",\n \"Among these genes, the expression level of a gene GenBank ID: AF055021, whose function was unknown, was increased by treatment with VEGF reaching the expression level of was 3.8 fold after 24 hours.\",\n \"Because a protein translated region was unclear from the information of sequence of AF055021 gene in a database of NCBI (National Center for Biotechnology Information), a base sequence of the same gene was searched with AssEst (maze) database and a consensus sequence of EST cluster was obtained.\",\n \"Based on this consensus sequence, homology search using Blastx database in AssEst revealed that it encoded KIAA1036 polypeptide (SEQ ID NO: 2) whose function was unknown.\",\n \"Additionally, it had 58% homology with a polypeptide of AK022567 whose function was unknown (FIG.\",\n \"1).\",\n \"KIAA1036 gene and AK022567 gene on NCBI database have open reading frames corresponding to 365 and 290 amino acid residues respectively, which are initiated from translation initiation codons and they are seem to be almost full length cDNA.\",\n \"A polypeptide of AK022567 was already published internationally as a secretory protein (WO99/3881).\",\n \"And KIAA1036 is predicted as a secretory protein because the homology of their amino acid sequences is 58%.\",\n \"Example 2 Expression of KIAA1036 in Human Normal Tissues Cloning of KIAA1036 gene from human umbilical vein vascular endothelial cell was tried.\",\n \"Two kinds of oligonucleotides were synthesized as primers for PCR on the basis of a nucleic acid sequence of 3′ untranslated region of KIAA1036 gene from NCBI database.\",\n \"Primer1: 5′-GTTCAGGACTGTCTTTCAGC-3′ (SEQ ID NO: 4) Primer2: 5′-GTCAATACTGATGGACTTGC-3′ (SEQ ID NO: 5) Total RNA was prepared according to an attached manual with RNeasy Midi (QIAGEN) from HUVEC which are human umbilical vein vascular endothelial cells.\",\n \"Single strand cDNA fragments were prepared from the obtained total RNA (10 μg) with SuperscriptII (Invitrogen).\",\n \"The obtained single strand cDNAs were used as templates and gene fragments were amplified with Primer1 (SEQ ID NO: 4) and Primer2 (SEQ ID NO: 5) by PCR.\",\n \"To one fiftieth of single strand cDNAs, was added TaKaRa Ex Taq (TaKaRa) (1.25 unit), Primer1 and Primer2 (1 μM each), a buffer (Mg2+ plus) for PCR reaction attached TaKaRa Ex Taq (5 μl) and dNTP Mixture whose final concentration was 200 μM.\",\n \"Distilled water was added to the mixture to be 50 μl in total.\",\n \"This was how samples for PCR were prepared.\",\n \"PCR reaction was held under the condition that is 94° C. for 3 minutes was once and the cycle (94° C. for 1 minute, 50° C. for 1 minute and 72° C. for 1 minute) was 30 times and then 72° C. for 5 minutes was once.\",\n \"The amplified cDNA fragment by the PCR reaction was separated by an agarose electrophoresis and 750 bp cDNA fragment (SEQ ID NO: 6) was collected.\",\n \"The obtained cDNA fragment was digested with restriction enzymes, ApaI, HincII and PstI and the lengths of cDNA fragments separated by an agarose gel electrophoresis were measured.\",\n \"And the collected cDNA fragment was confirmed as a target.\",\n \"The cDNA fragment (50 ng) amplified with Primer1 and Primer2 was labeled with 32P using [α-32P] dCTP (3000 Ci/m mol, 10 mCi/ml) (Amersham Pharmacia Bioteque) by an attached manual with Rediprime II DNA Labeling System (Amersham Pharmacia Bioteque).\",\n \"Northern blot was held with these cDNA fragments labeled with 32P as a probe.\",\n \"Nylon membrane of Multiple Tissue Northern (MTN™) Blots Human 12-Lane MTN Blot (Clontech) was immersed with GMC buffer (including 0.5M phosphoric acid buffer, pH 7.2, 1% cattle serum albumin, 1 mM EDTA and 7% SDS) and prehybridization was held at 65° C. for 1 hour.\",\n \"After that, the nylon membrane was immersed with GMC buffer containing probes labeled with 32P and hybridization was held at 65° C. for 16 hours.\",\n \"After hybridization, the nylon membrane was washed with 2×SSC (1×SSC was 15 mM sodium citrate, 150 mM NaCl, pH 7.0) at a room temperature for 5 minutes, 1×SSC (including 0.2% SDS) at 50° C. for 30 minutes and 0.5×SSC (including 0.2% SDS) at 50° C. for 30 minutes one after another and dried with air.\",\n \"Subsequently, this nylon membrane was put on an imaging plate cassette (FUJI FHOTO FILM CO., LTD.) with an imaging plate (FUJI FHOTO FILM CO., LTD.).\",\n \"Image analysis was held with image analyzer FLA3000 (FUJI FHOTO FILM CO., LTD.).\",\n \"As a result, signals which showed about 6 kb length were detected from brain, heart, skeletal muscle, spleen, kidney, liver, small intestine and placenta and the strong expression was selectively observed in brain and placenta.\",\n \"Example 3 Expression of KIAA1036 in Human Umbilical Vein Endothelial Cells As the VEGF-induced expression of KIAA1036 was originally found by Gene Expression Microarray Analysis using DNA chip as described in Example 1, the result was confirmed by Northern blot with KIAA1036 probe and total RNA prepared from VEGF-treated HUVEC.\",\n \"Endocell HUVEC (KURABO) which are human umbilical vein endothelial cells were cultured in M199 medium containing 1 nM human VEGF165 (R&D Systems) and 1% FCS as described in Example 1.After 0, 0.5, 1, 2, 6, 12 and 24 hours of culture, total RNA was prepared from these cells according to an attached manual with RNeasy Midi (QIAGEN).\",\n \"After electrophoresis of each RNA sample (5 μg), the samples were transferred to a nylon membrane, Hybond-XL (Amersham Pharmacia Bioteque).\",\n \"That nylon membrane was immersed with GMC buffer and prehybridization was held at 65° C. for 1 hour.\",\n \"After prehybridization, the nylon membrane was immersed with GMC buffer containing probes labeled with 32P as Example 2 and hybridization was held at 65° C. for 16 hours.\",\n \"After hybridization, the nylon membrane was washed with 2×SSC at a room temperature for 5 minutes, 1×SSC (including 0.2% SDS) at 50° C. for 30 minutes and 0.5×SSC (including 0.2% SDS) at 50° C. for 30 minutes one after another and dried with air.\",\n \"Subsequently, this nylon membrane was put on an imaging plate cassette (FUJI FHOTO FILM CO., LTD.) with an imaging plate (FUJI FHOTO FILM CO., LTD.).\",\n \"Image analysis was held with image analyzer FLA3000 (FUJI FHOTO FILM CO., LTD.).\",\n \"Next, probes were dehybridized by immersing the same nylon membrane in a bath with boiling water and Northern blot was held with human β-actin cDNA fragments labeled with 32P as Example 2 as probes.\",\n \"Signal intensities of KIAA1036 gene were normalized with that of β-actin gene at each time point.\",\n \"And then a ratio of the expression level of KIAA1036 in each time point to 0 hour was calculated.\",\n \"As a result, the expression was inhibited at 0.5 to 2 hours after adding VEGF but a high gene expression, which was 3.2 fold, was induced after 24 hours.\",\n \"This result was same as that obtained by DNA chip (FIG.\",\n \"2).\",\n \"Example 4 Expression of KIAA1036 in Human Cancer Tissues The difference of expression levels of the genes in a normal tissue and a cancer tissue of all kinds of organ from human was analyzed.\",\n \"Matched Tumor/Normal Expression Array (Clontech), which is a nylon membrane with cDNAs synthesized from a cancer tissue and a normal tissue of chest, uterus, colon, stomach, ovary, lung, kidney, rectum, cervix, small intestine or prostate of the same patient, was immersed with GMC buffer containing probes labeled with 32P as Example 2 and hybridization was held at 65° C. for 16 hours.\",\n \"After hybridization, the nylon membrane was washed with 2×SSC at a room temperature for 5 minutes, 1×SSC (including 0.2% SDS) at 50° C. for 30 minutes and 0.5×SSC (including 0.2% SDS) at 50° C. for 30 minutes one after another and dried with air.\",\n \"Subsequently, this nylon membrane was put on an imaging plate cassette (FUJI FHOTO FILM CO., LTD.) with an imaging plate (FUJI FHOTO FILM CO., LTD.).\",\n \"Image analysis was held with image analyzer FLA3000 (FUJI FHOTO FILM CO., LTD.).\",\n \"After hybridization of the nylon membrane, signal intensities were measured with β-actin genes labeled with 32P as probes as described in Example 3.Signal intensities of KIAA1036 was normalize with signal intensities by β-actin probe in each organ, and a ratio of cancer tissues to normal tissues in the same patient was calculated.\",\n \"Then a change of an expression level of the genes in all kinds of cancer tissue was analyzed.\",\n \"As a result, expression of KIAA1036 genes were increased in cancer tissues of all 4 patients with ovarian cancer and 11 patients with colon cancer patient (FIG.\",\n \"3).\",\n \"Example 5 Expression of KIAA1036 in COS Cells cDNA encoding the polypeptide that 3×FLAG sequence was added to a carboxy-terminal of KIAA1036 amino acid sequence as set forth in Met of 1-Val of 365 of SEQ ID NO: 2 was inserted to a expression vector in animal cells and expressed in COS cells.\",\n \"A base sequence as set forth in A of 386-C of 1480 of SEQ ID NO: 1 encoding KIAA1036 was inserted in the NotI/XbaI sites of an expression vector in animal cells, p3×FLAG-CMV-13 and p3×FLAG-CMV-14(SIGMA-Aldrich) and plasmids, pFLAG13-1036 and pFLAG14-1036 which express polypeptides that 3×FLAG was added to carboxy- or amino-terminals of KIAA1036, respectively, were constructed.\",\n \"COS-7 cells (African green monkey kidney cells (SV40-transformed cells)) were used as host of pFLAG13-1036 and pFLAG14-1036 and transformation was operated by an attached manual with FuGENE6 reagent (Roche Diagnostics).\",\n \"COS-7 cells (5×105 cell/well in 60 mm dishes) were cultured in D-MEM medium containing 10% FCS at 37° C. for 16 hours.\",\n \"After medium was changed, pFLAG13-1036 (2 μg) and FuGENE6 (10 μl) were added to the cells and the cells were cultured for 48 hours.\",\n \"pFLAG14-1036 and p3×FLAG-CMV-14 without KIAA1036 cDNA as a control were also used in a same method to make transfectants and the transfectants were cultured.\",\n \"Expression of KIAA1036 polypeptide fused with FLAG was analyzed by Western blotting, a method of Laemli.\",\n \"The culture supernatants (10 μl) of all kinds of COS-7 transfectants were subjected to SDS-PAGE and electrically transfered to a nitrocellulose membrane in a nitrocellulose membrane kit (TEFCO).\",\n \"The nitrocellulose membrane was immersed with a blocking reagent (TBS, pH 7.6; including 5%(w/v) skim milk) and shook at a room temperature for 1 hour.\",\n \"And the membrane was immersed in TBS reagent (pH7.6; including 5%(w/v) skim milk) containing anti-FLAG M2 antibody labeled with peroxidase (HRP) (SIGMA-Aldrich), put static at a room temperature for 1 hour.\",\n \"Then the nitrocellulose membrane was washed with a washing reagent (TBS, pH7.6; including 0.05%(v/v) TritonX-100) for 10 minutes 3 times.\",\n \"The nitrocellulose membrane and ECL Plus Western Blotting Detection Reagents (Amersham Pharmacia Bioteque) were reacted at a room temperature.\",\n \"The nitrocellulose membrane and X-ray film X-OMAT AR (Kodak) were put on X-OMAT cassette (Kodak) immediately and exposed for a few minutes.\",\n \"As a result, expected bands, which show about 50 kDa, from the culture supernatant of COS cells transfected with pFLAG13-1036 and pFLAG14-1036 were detected but the band was not detected in a control.\",\n \"Therefore, it was confirmed that KIAA1036 gene encodes a KIAA1036 protein, which is able to express, and that KIAA1036 was a secretory protein in animal cells.\",\n \"Example 6 Expression of KIAA1036 Protein in a Baculovirus System Expressions in a baculovirus system were operated with Bac-to-Bac Baculovirus Expression System (Invitrogen) according to an attached manual.\",\n \"cDNA fragments encoding KIAA1036 with 3×FLAG at the carboxy-terminal described in Example 5 was inserted in NotI/XhoI sites of a plasmid pFastBac1, an expression plasmid for insect cells, to make pFASTBac1036.The recombinant plasmid was introduced in Max Efficiency DH10 Bac Competent Cells and cDNA was transferred into baculovirus genome (bacmid DNA) to give rise to a recombinant bacmid DNA.\",\n \"The recombinant bacmid DNA was introduced in Sf9 (ATCC: CRL-1711, ovary cell) cell with CellFECTIN Reagent (Invitrogen) to obtain a recombinant baculovirus.\",\n \"A culture supernatant (0.5 ml) of Sf9 transfectants was added into a culture medium (50 ml) of Sf9 cells with vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or malignant neoplasma (The concentration was 1.5×106 cells/ml) and cultured at 28° C. for 96 hours.\",\n \"The transformed cells were collected by centrifugation.\",\n \"Expression of the FLAG-tagged KIAA1036 was confirmed by Western blotting as described in Example 5.The collected transformants were washed once with PBS (5 ml, pH7.4).\",\n \"After washing, the cells were suspended in Lysis reagent (5 ml) (including 50 mM Tris-HCl, pH7.4; 0.15M NaCl, 0.1 mM EDTA/2Na, 0.1 mM EGTA, 1 mM DTT, 0.1 mM Amidinophenyl methansulfonyl fluride hydrochloride or 0.1% NP-40), treated with ultrasonic waves on ice for 15 seconds for 4 times by MICROCON (HEART SYSTEMS) and separated by centrifugation at 14,500×g for 20 minutes.\",\n \"The same amount of Lysis reagent was added to the supernatant to make it a crude protein solution.\",\n \"After a crude protein solution (10 μl) was subjected to SDS-PAGE, it was electrically transferred on a nitrocellulose membrane in nitrocellulose membrane kit (TEFCO).\",\n \"The nitrocellulose membrane was immersed in a blocking reagent (TBS, pH7.6; including 5%(w/v) skim milk) and shook at a room temperature for 1 hour.\",\n \"After that, it was immersed in a TBS reagent (pH7.6; including 5%(w/v) skim milk) containing anti-FLAG M2 antibody labeled with peroxidase (HRP) (SIGMA-Aldrich), put static at a room temperature for 1 hour.\",\n \"And then the nitrocellulose membrane was washed with the washing reagent (TBS, pH7.4; including 0.05%(v/v) TritonX-100) for 10 minutes 3 times.\",\n \"The nylon membrane and ECL Plus Western Blotting Detection Reagents (Amersham Pharmacia Bioteque) were reacted at a room temperature.\",\n \"The nitrocellulose membrane and X-ray hyper film ECL (Amersham Pharmacia Bioteque) were put on X-OMAT cassette Kodak) immediately and exposed for a few minutes.\",\n \"KIAA1036 FLAG proteins were purified by an affinity chromatography with anti-FLAG antibodies.\",\n \"After an ANTI-FLAG M2-Agarose (SIGMA-Aldrich) affinity column was equilibrated with TBS, pH7.4 (including 0.1% NP-40), a crude protein solution was added and FLAG proteins were separated.\",\n \"The columns were then washed with TBS (pH7.4; including 0.1% NP-40) and the protein was eluted with Gly-HCl (pH3.5; including 0.1% NP-40) to be 1 ml/fraction.\",\n \"Each fraction was neutralized with 1M Tris-HCl (pH8.0; 20 μl) to make it an affinity purification fraction.\",\n \"A crude protein fraction (10 μl) and an affinity purified fraction (10 μl) were subjected to SDS-PAGE and the gel was stained by Bio-Safe Coomassie (BIO-RAD).\",\n \"As a result, an expected band with about 50 kDa was detected.\",\n \"Furthermore, after SDS-PAGE, the purified fraction was electrically transferred to PVDF membrane (BIO-RAD) and a band stained with Bio-Safe Coomassie (BIO-RAD) was cut off, and then a sequence of an amino terminal was analyzed by an amino acid sequencer, Procise Model 491 (Applied Biosystems).\",\n \"As a result, an amino acid sequence as set forth in Pro of 2-Gly of 10 of SEQ ID NO: 2 was detected and it was confirmed that the expressed protein was KIAA1036.Example 7 A Blocking Effect of DNA Synthesis by KIAA1036 in Vascular Endothelial Cells To examine an effect of KIAA1036 protein to the proliferation of vascular endothelial cells, BrdU (bromodeoxyuridine) incorporation was measured in HUVEC cells treated with VEGF in the presence (a treated group) or the absence (a control group) of KIAA1036.A KIAA1036 protein, which was expressed by a baculovirus expression system in Example 6 and purified by an affinity chromatography, was used.\",\n \"Incorporation of BrdU into cells was measured with cell propagation ELISA and a BrdU chemiluminescence kit according to an attached manual.\",\n \"Endocell HUVEC (KURABO) which are human vascular endothelial cells (3000 cells/plate) were cultured on collagen-coated plates (IWAKI) with M199 medium (Nissui) containing 5% fetal calf serum (FCS) at 37° C. for 24 hours.\",\n \"For the VEGF-treated group, medium was changed to M199 medium including 1% FCS, 1 nM human VEGF165 (R&D Systems) and 0, 1, 10 nM KIAA1036 protein.\",\n \"For the VEGF-untreated group (control group) medium was changed to the same mediums without human VEGF165 containing 0, 1 or 10 nM KIAA1036 protein.\",\n \"A solution labeled with BrdU (0.02 ml/well) was added and the cells were cultured at 37° C. for 12 hours.\",\n \"The cultured cells were put static at a room temperature for 1 hour to immobilize and an anti-BrdU antibody solution labeled with POD (0.1 ml/well) was added and the cells were put static at a room temperature for 1 hour.\",\n \"The cells were washed with a washing buffer for 5 minutes 3 times and a luminecsence substrate (0.1 ml/well) was added.\",\n \"After 3 minutes of shaking, the instantaneous fluorescence for 5 seconds was measured by Luminescencer JNR (ATTO) within 10 minutes.\",\n \"As a result, VEGF promoted DNA synthesis of HUVEC, but KIAA1036 protein doses-dependently inhibited it (FIG.\",\n \"4).\",\n \"Example 8 A Blocking Effect to the Migration of Vascular Endothelial Cells by KIAA1036 Protein An effect to the migration of vascular endothelial cells of KIAA1036 protein was examined by a modified Boyden chamber method.\",\n \"To the lower chambers of Transwell (Corning coaster), M199 medium (600 μl /well) containing 0.25 nM human VEGF165 (R&D Systems) and 1 or 10 nM KIAA1036 protein was added for samples treated with VEGF (test group), and M199 medium (600 μl/well) containing 1 or 10 nM KIAA1036 protein without VEGF was added for a control group.\",\n \"Endocell HUVEC (KURABO) (4×104 cells/well) were cultured in M199 medium containing 5% FCS for 16 hours and then seeded in the upper chambers and cultured at 37° C. for 4 hours.\",\n \"Filters of Transwell were immobilized with Diffquick fix reagent (Sysmex) and then stained with Diffquick stain (Sysmex).\",\n \"After wiping the side of the upper chamber of filter, the cell migration ability toward VEGF was measured by counting the number of the cells migrating to the lower part.\",\n \"As a result, VEGF promoted migration of HUVEC, but KIAA1036 protein significantly inhibited it (FIG.\",\n \"5).\",\n \"Example 9 A Blocking Effect to Network Formation of Vascular Endothelial Cells by KIAA1036 Protein To examine an effect to network formation of vascular endothelial cells by KIAA1036 protein, network formation of HUVEC in the presence (treated group) or the absence (control group) of KIAA1036 protein was observed.\",\n \"Matrigel (Becton Dickinson Biosciences) (1 ml/well) was divided in 6-well tissue culture plates chilled on ice and put static at 37° C. for 1 hour to be gelatinized.\",\n \"Endocell HUVEC (KURABO) were suspended in EBM medium containing 10% FCS and poured onto the matrigel at 5×104 cells/well with a final concentration of KIAA1036 protein at 1 or 10 nM (test group) or without KIAA1036 protein.\",\n \"After 9 hours of incubation at 37° C., the network formations of HUVEC in test divisions and control divisions were observed with an inverted microscope.\",\n \"As a result, it was confirmed that KIAA1036 protein obviously inhibits the network formation.\",\n \"Industrial Applicability The present invention provides with a detection method of mRNA of KIAA1036, a detection method of a polypeptide of KIAA1036, a marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease or malignant neoplasm, a diagnostic kit and a therapeutic agent.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476985"}}},{"rowIdx":991,"cells":{"text":{"kind":"list like","value":[["Message authentication","There is disclosed a technique of providing message authentication in a communication system comprising the steps of: transmitting a first message from a first device to a second device; transmitting a second message from the second device to the first device, the second message including a message authentication code determined using said first and second messages; transmitting a third message from the first device to the second device, the third messages including a message authentication code determined using the third message.","The message authentication code of the third message may be additionally based on the second or the second and first messages."],["1.A method of providing message authentication in a communication system comprising the steps of: transmitting a first message from a first device to a second device; transmitting a second message from the second device to the first device, the second message including a message authentication code determined using said first and second messages; transmitting a third message from the first device to the second device, the third message including a message authentication code determined using the third message.","2.A method according to claim 1, further comprising the step of storing the first message in the first device.","3.A method according to claim 1 further comprising the steps of: responsive to receipt of the second message, determining an expected message authentication code using said first and second messages; and comparing the expected message authentication code to the received message authentication code.","4.A method according to claim 3 further comprising the step of discarding the second message if the expected message authentication code does not match the received message authentication code.","5.A method according to claim 1 further comprising the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message; and comparing the expected message authentication code to the received message authentication code.","6.A method according to claim 5 further comprising the step of discarding the third message if the expected message authentication code does not match the received message authentication code.","7.A method according to claim 1, wherein the third message includes a message authentication code determined using the third message and the second message.","8.A method according to claim 7, further comprising the step of storing the second message in the second drive.","9.A method according to claim 7, further comprising the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message and said second message; and comparing the expected message authentication code to the received message authentication code.","10.A method according to claim 9 further comprising the step of discarding the third message if the expected message authentication code does not match the received message authentication code.","11.A method according to claim 1, wherein the third message includes a message authentication determined using the third message, the second message, and the first message.","12.A method according to claim 11, further comprising the step of storing said first message in the second device.","13.A method according to claim 11, further comprising the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message, said second message, and said first message; and comparing the expected message authentication code to the received message authentication code.","14.A method according to claim 13 further comprising the step of discarding the third message if the expected message authentication code does not match the received message authentication code.","15.A method according to claim 1 wherein the first and second devices are elements of a mobile communication system.","16.A method of providing message authentication in a communication system comprising transmitting a plurality of messages between a first device and a second device, wherein a message includes a message authentication code determined using at least said message and another message includes a further message authentication code determined using at least said other message, whereby the number of required message authentication code computations and transmissions may be reduced to two independently of the number of steps of the authentication procedure.","17.A communication system for providing message authentication between two communicating devices, each communicating device having respective transmitting and receiving means, and each communicating device having means for generating a message authentication code, comprising: in the first device, transmitting a first message from the second device; in the second device, transmitting a second message to the first device, the second message including a message authentication code determined using said first and second messages; in the first device transmitting a third message to the second device, the third messages including a message authentication code determined using the third message.","18.A communication system according to claim 17, further including, in the first device: responsive to receipt of the second message, determining an expected message authentication code using said first and second messages; and comparing the expected message authentication code to the received message authentication code.","19.A communication system according to claim 17 wherein the second message is discarded if the expected message authentication code does not match the received message authentication code.","20.A communication system according to claim 17 further comprising, in the second device: responsive to receipt of the third message, determining an expected message authentication code using said third message; and comparing the expected message authentication code to the received message authentication code.","21.A communication system according to claim 20 wherein the third message is discarded if the expected message authentication code does not match the received message authentication code.","22.A communication system according to claim 17, wherein the first device generates the third message including a message authentication code determined using the third message and the second message.","23.A communications system according to claim 22, wherein responsive to receipt of the third message, the second device determines an expected message authentication code using said third message and said second message; and compares the expected message authentication code to the received message authentication code.","24.A communications system according to claim 23 wherein the third message if the expected message authentication code does not match the received message authentication code.","25.A communications system according to claim 17 wherein the third message generated by the first device includes a message authentication code determined using the third message, the second message, and the first message.","26.A communication system according to claim 25 wherein responsive to receipt of the third message, the second device determines an expected message authentication code using said third message, said second message, and said first message; and compares the expected message authentication code to the received message authentication code.","27.A communication system according to claim 26 wherein the second device discards the third message if the expected message authentication code does not match the received message authentication code.","28.A communication system according to claim 17 wherein the first and second device are elements of a mobile communication system.","29.A communication system according to claim 28, wherein the first and second devices are one of a mobile terminal and a network element.","30.A communication system according to claim 28, wherein the mobile communication system comprises a GERAN system."],["<SOH> BACKGROUND OF THE INVENTION <EOH>All telecommunication is subject to the problem of how to make sure that the information received has been sent by an authorized sender and not by somebody who is trying to masquerade as the sender.","The problem is particularly evident in cellular telecommunication systems, where the air interface presents a potential platform for eavesdropping and replacing the contents of a transmission by using higher transmission levels, even from a distance.","A basic solution to this problem is the authentication of the communicating parties.","An authentication process aims to discover and check the identity of both the communicating parties, so that each party receives information about the identity of the other party and can rely on the identification to a sufficient degree.","Authentication is typically performed in a specific procedure at the beginning of a connection.","However, this does not adequately protect subsequent messages from unauthorized manipulation, insertion, and deletion.","Thus, there is a need for the separate authentication of each transmitted message.","The latter task can be carried out by appending a message authentication code (MAC-I) with a particular value to the message at the transmitting end and checking the MAC-I value at the receiving end.","A MAC-I is typically a relatively short string of bits based in some specified way on the message it protects and on a secret key known both by the sender and by the recipient of the message.","The secret key is generated and agreed on typically in connection with the authentication procedure at the beginning of the connection.","In some cases the algorithm that is used to calculate the MAC-I based on the secret key and on the message may also be secret.","The process of authentication of single messages is often called integrity protection.","To protect the integrity of signaling, the transmitting party computes a MAC-I value based on the message to be sent and the secret key using the specified algorithm, and sends the message with the MAC-I value.","The receiving party recomputes a MAC-I value based on the message and the same secret key according to the same specified algorithm, and compares the received MAC-I and the calculated MAC-I.","If the two MAC-I values match, the recipient can trust that the message is intact and has been sent by the authorized party.","Known integrity protection schemes are not completely reliable.","A third party can attempt to manipulate a message transmitted between a first and a second party.","There are two main alternative methods for forging a MAC-I value for a modified or a new message: obtaining the secret key; or by trying directly without the secret key.","The secret key can be obtained by a third party in two ways: i) by computing all possible keys until a key is found matching the data of observed message MAC-I pairs, or by otherwise breaking the algorithm for producing MAC-I values; or ii) by directly capturing a stored or transmitted secret key.","The original communicating parties can prevent a third party from obtaining the secret key by using an algorithm which is cryptographically strong and which uses a secret key which is long enough to prevent the exhaustive search of all keys, and by using other security means for the transmission and storage of secret keys.","A third party may try to disrupt the sending of messages between the two parties without a secret key by guessing the correct MAC-I value, or by replaying some earlier message transmitted between the two parties for which message the correct MAC-I is known from the original transmission.","Guessing of the correct MAC-I value can be made difficult by using long MAC-I values.","The MAC-I value should be long enough to reduce the probability of correct guessing to a sufficiently low level compared to the benefit gained by one successful forgery.","For example, using a 32 bit MAC-I value reduces the probability of a correct guess to 1/4294967296, which is small enough for most applications.","Obtaining a correct MAC-I value using the replay attack, i.e.","by re-playing an earlier message, can be prevented by introducing a varying parameter to the calculation of the MAC-I values.","For example, a time stamp value, a sequence number, or a random number can be used as further input to the MAC-I algorithm, in addition to the secret integrity key and the message.","In the following, the prior art methods are described in more detail.","When using sequence numbers, each party has to keep track of which sequence numbers have already been used and are not acceptable any more.","The easiest way to implement this is to store the highest sequence number used in MAC-I calculations so far.","This approach has the drawback that between connections each party must maintain state information which is at least to some level synchronized.","That is, they need to store the highest sequence number used so far.","This requires the use of a large database in the network.","A further approach is to include a random number in each message, which the other side must use in MAC-I calculation the next time a message is sent for which MAC-I authentication is required.","This approach has the same drawback as the previous one, i.e.","between connections each party must maintain state information, which requires the use of a large database in the network.","By way of example, FIG.","1 illustrates the computation of a message authentication code in the UTRAN (UMTS Terrestrial Radio Access Network), which is a wideband multiple access radio network currently being specified in the 3GPP (Third Generation Partnership Project).","The length of the MAC-I used in UTRAN is 32 bits.","Block 100 represents the UMTS integrity algorithm for generating the message authentication code.","The UMTS integrity algorithm used in block 100 is a one-way cryptographic function for calculating the 32 bit Message Authentication Code (MAC-I) based on the five input parameters shown in FIG.","1 .","A one-way function makes it impossible to derive the unknown input parameters from a MAC-I, even if all but one input parameter are known.","The input parameters for calculating the MAC-I 20 are: the actual signaling message 10 (after encoding) to be sent, a secret integrity key 12 , a number COUNT-I value 14 for the message to be integrity protected, a value indicating the direction of transmission 16 , (i.e.","whether the message is sent in uplink or downlink direction), and a random number 18 (FRESH) generated by the network.","The COUNT-I value consists of a hyper frame number HFN and the message sequence number SN.","The computing block 100 calculates the message authentication code by applying the afore-mentioned parameters to the integrity algorithm, which is called the f9 algorithm in 3GPP Release'99 specifications.","FIG.","2 illustrates a typical message to be sent over a radio interface.","The message is a layer N protocol data unit (PDU) 200 , which is transferred as a payload 30 in a layer N- 1 PDU 201 .","In the present example, layer N represents the Radio Resource Control (RRC) protocol in the radio interface and layer N- 1 represents the Radio Link Control (RLC) layer.","The layer N- 1 PDU normally has a fixed size, which depends on the physical layer (the lowest layer, not visible in FIG.","2 ) the channel type used and on other parameters, e.g.","modulation, channel coding, interleaving.","If layer N PDUs are not exactly the size of the payload 30 offered by layer N- 1 as is normally the case, layer N- 1 can utilize functions like segmentation, concatenation, and padding to make layer N- 1 PDUs a fixed size.","In the example discussed herein, a layer N PDU consisting of the actual signaling data 22 and the Integrity Check Info is discussed.","The Integrity Check Info consists of the 32 bit MAC-I 26 and the message sequence number SN 24 , which is needed at the peer end for the recalculation of MAC-I.","The total length of the message 200 is then a combination of the signaling bits and the Integrity Check Info bits.","At the receiving end the message is received including the signaling data part and the Integrity Check Info (which comprises the message sequence number SN and the 32-bit MAC-I).","The signaling data together with the Integrity Check Info (ie the secret integrity the COUNT-I value, the direction of transmission, and the random number (FRESH)), are processed in a computing block, for example a function like the UTRAN f9 function, with a fixed function.","A thus generated received computed message authentication code XMAC-I is then compared with the message authentication code MAC-I received in the transmitted message.","If the two codes match (ie XMAC-I matches MAC-I), the recipient can trust that the message is intact, and the recipient then accepts the message.","Otherwise, the message is discarded.","The frame dependent COUNT-I number is actually the sum of a locally generated and incremented frame number HFN (Hyper Frame Number), which is added to the message sequence number, for example RRC_SN, and included in the message.","The HFN is incremented each time SN reaches its maximum value (SN is normally very short, e.g.","4 bits).","As mentioned hereinabove the transmitted.block (layer N- 1 PDU) normally has a fixed length.","However, it may be that the signaling data bits together with the Integrity Check Info require more space than that provided in one layer N- 1 PDU payload.","One known way to deal with this problem is to segment the signaling message.","In segmentation a signaling message, which is too long to fit in a single layer N- 1 block, is passed on to a lower layer, where it is split up into two blocks (two layer N- 1 PDUs), each with an appropriate layer N- 1 header.","Two blocks is just an example here, naturally a larger message may require even more than two blocks.","If the second layer N- 1 PDU is not totally filled with the layer N data, padding bits are inserted.","At the receiving end before transferring to a higher layer, the two layer N- 1 payloads are reassembled into one layer N PDU.","To a person skilled in the art, it is immediately obvious that the use of padding bits is a potential waste of resources.","TDMA systems, for example, have a limited radio block size, whereby a message including the full message authentication code does not necessarily fit into one radio block.","This leads to the difficulty that the message has either to be sent without the MAC-I or in one or more additional segments.","In addition, there are certain time critical messages, for example, handover messages, which must be sent in one radio block only.","Generally, segmentation is not desirable, because it wastes radio resources and slows down the signaling procedure unnecessarily.","One way to solve the above problem is to make the length of the message authentication code shorter than 32 bits.","It has been proposed that such a message should include a field that defines the length of the message authentication code, a two-bit identifier, for example.","This identifier allows certain discrete values: 8, 16, 24 and 32.This solution still has some problems.","First, the identifier always takes two extra bits from the length of the message.","Second, the discrete values are not flexible, and in some cases this can lead to the same problem as above, i.e.","segmentation is needed for certain messages.","A particularly advantageous technique for addressing the above stated problems is disclosed in Finnish patent application number FI20002453.This discloses a technique that allows the transmission of a message in a single lower layer data block even when the length of the message including the integrity check info exceeds the length of the lower layer data block.","In all current known solutions, each message is authenticated separately.","Each message contains a sequence number of the protocol it belongs to.","Hence a binding between different steps in the existing solutions is achieved through the sequence number.","The drawback to existing solutions is that at each step a 32-bit MAC is transmitted which takes a significant part of the signaling bandwidth.","It is therefore an object to the present invention to provide an improved technique for taking the integrity of messages."],["<SOH> SUMMARY OF THE INVENTION <EOH>In accordance with the present invention there is provided a method of providing message authentication in a communication system comprising the steps of: transmitting a first message from a first device to a second device; transmitting a second message from the second device to the first device, the second message including a message authentication code determined using said first and second messages; transmitting a third message from the first device to the second device, the third messages including a message authentication code determined using the third message.","In order to minimize the risk of ‘replay attacks’, the sequence numbers should preferably be maintained in the protocol.","This is particularly important if the messages are short or otherwise small in number.","The method may further comprise the step of storing the first message in the first device.","The method may further comprise the steps of: responsive to receipt of the second message, determining an expected message authentication code using said first and second messages; and comparing the expected message authentication code to the received message authentication code.","The method may further comprise the step of discarding the second message if the expected message authentication code does not match the received message authentication code.","The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message; and comparing the expected message authentication code to the received message authentication code.","The method may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.","The third message may include a message authentication code determined using the third message and the second message.","The method may further comprise the step of storing the second message in the second device.","The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message and said second message; and comparing the expected message authentication code to the received message authentication code.","The method according to claim 9 may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.","The third message may include a message authentication code determined using the third message, the second message, and the first message.","The method may further comprise the step of storing said first message in the second device.","The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message, said second message, and said first message; and comparing the expected message authentication code to the received message authentication code.","The method may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.","The invention may thus advantageously reduce the bandwidth used for authentication.","In a multi-step protocol, the invention provides for the messages exchanged at different steps to be grouped together for message authentication code computations.","A message is saved by the sending party for subsequent verification at a later step.","The number of required message authentication code computations and transmissions may be reduced to two independently of the number of steps in the authentication procedure.","In three step signaling procedures, there is thus no need for a message authentication code to be computed in the first step.","The invention is further advantageously applicable to procedures containing more than three messages.","All communications that occur between two parties in various steps may be authenticated by the communicating parties in the last two steps of the protocol.","The first and second devices are preferably elements of a mobile communication system."],["FIELD OF THE INVENTION The present invention generally relates to a method for checking the integrity of messages in a communication system, particularly but not exclusively between a mobile station and a cellular network.","BACKGROUND OF THE INVENTION All telecommunication is subject to the problem of how to make sure that the information received has been sent by an authorized sender and not by somebody who is trying to masquerade as the sender.","The problem is particularly evident in cellular telecommunication systems, where the air interface presents a potential platform for eavesdropping and replacing the contents of a transmission by using higher transmission levels, even from a distance.","A basic solution to this problem is the authentication of the communicating parties.","An authentication process aims to discover and check the identity of both the communicating parties, so that each party receives information about the identity of the other party and can rely on the identification to a sufficient degree.","Authentication is typically performed in a specific procedure at the beginning of a connection.","However, this does not adequately protect subsequent messages from unauthorized manipulation, insertion, and deletion.","Thus, there is a need for the separate authentication of each transmitted message.","The latter task can be carried out by appending a message authentication code (MAC-I) with a particular value to the message at the transmitting end and checking the MAC-I value at the receiving end.","A MAC-I is typically a relatively short string of bits based in some specified way on the message it protects and on a secret key known both by the sender and by the recipient of the message.","The secret key is generated and agreed on typically in connection with the authentication procedure at the beginning of the connection.","In some cases the algorithm that is used to calculate the MAC-I based on the secret key and on the message may also be secret.","The process of authentication of single messages is often called integrity protection.","To protect the integrity of signaling, the transmitting party computes a MAC-I value based on the message to be sent and the secret key using the specified algorithm, and sends the message with the MAC-I value.","The receiving party recomputes a MAC-I value based on the message and the same secret key according to the same specified algorithm, and compares the received MAC-I and the calculated MAC-I.","If the two MAC-I values match, the recipient can trust that the message is intact and has been sent by the authorized party.","Known integrity protection schemes are not completely reliable.","A third party can attempt to manipulate a message transmitted between a first and a second party.","There are two main alternative methods for forging a MAC-I value for a modified or a new message: obtaining the secret key; or by trying directly without the secret key.","The secret key can be obtained by a third party in two ways: i) by computing all possible keys until a key is found matching the data of observed message MAC-I pairs, or by otherwise breaking the algorithm for producing MAC-I values; or ii) by directly capturing a stored or transmitted secret key.","The original communicating parties can prevent a third party from obtaining the secret key by using an algorithm which is cryptographically strong and which uses a secret key which is long enough to prevent the exhaustive search of all keys, and by using other security means for the transmission and storage of secret keys.","A third party may try to disrupt the sending of messages between the two parties without a secret key by guessing the correct MAC-I value, or by replaying some earlier message transmitted between the two parties for which message the correct MAC-I is known from the original transmission.","Guessing of the correct MAC-I value can be made difficult by using long MAC-I values.","The MAC-I value should be long enough to reduce the probability of correct guessing to a sufficiently low level compared to the benefit gained by one successful forgery.","For example, using a 32 bit MAC-I value reduces the probability of a correct guess to 1/4294967296, which is small enough for most applications.","Obtaining a correct MAC-I value using the replay attack, i.e.","by re-playing an earlier message, can be prevented by introducing a varying parameter to the calculation of the MAC-I values.","For example, a time stamp value, a sequence number, or a random number can be used as further input to the MAC-I algorithm, in addition to the secret integrity key and the message.","In the following, the prior art methods are described in more detail.","When using sequence numbers, each party has to keep track of which sequence numbers have already been used and are not acceptable any more.","The easiest way to implement this is to store the highest sequence number used in MAC-I calculations so far.","This approach has the drawback that between connections each party must maintain state information which is at least to some level synchronized.","That is, they need to store the highest sequence number used so far.","This requires the use of a large database in the network.","A further approach is to include a random number in each message, which the other side must use in MAC-I calculation the next time a message is sent for which MAC-I authentication is required.","This approach has the same drawback as the previous one, i.e.","between connections each party must maintain state information, which requires the use of a large database in the network.","By way of example, FIG.","1 illustrates the computation of a message authentication code in the UTRAN (UMTS Terrestrial Radio Access Network), which is a wideband multiple access radio network currently being specified in the 3GPP (Third Generation Partnership Project).","The length of the MAC-I used in UTRAN is 32 bits.","Block 100 represents the UMTS integrity algorithm for generating the message authentication code.","The UMTS integrity algorithm used in block 100 is a one-way cryptographic function for calculating the 32 bit Message Authentication Code (MAC-I) based on the five input parameters shown in FIG.","1.A one-way function makes it impossible to derive the unknown input parameters from a MAC-I, even if all but one input parameter are known.","The input parameters for calculating the MAC-I 20 are: the actual signaling message 10 (after encoding) to be sent, a secret integrity key 12, a number COUNT-I value 14 for the message to be integrity protected, a value indicating the direction of transmission 16, (i.e.","whether the message is sent in uplink or downlink direction), and a random number 18 (FRESH) generated by the network.","The COUNT-I value consists of a hyper frame number HFN and the message sequence number SN.","The computing block 100 calculates the message authentication code by applying the afore-mentioned parameters to the integrity algorithm, which is called the f9 algorithm in 3GPP Release'99 specifications.","FIG.","2 illustrates a typical message to be sent over a radio interface.","The message is a layer N protocol data unit (PDU) 200, which is transferred as a payload 30 in a layer N-1 PDU 201.In the present example, layer N represents the Radio Resource Control (RRC) protocol in the radio interface and layer N-1 represents the Radio Link Control (RLC) layer.","The layer N-1 PDU normally has a fixed size, which depends on the physical layer (the lowest layer, not visible in FIG.","2) the channel type used and on other parameters, e.g.","modulation, channel coding, interleaving.","If layer N PDUs are not exactly the size of the payload 30 offered by layer N-1 as is normally the case, layer N-1 can utilize functions like segmentation, concatenation, and padding to make layer N-1 PDUs a fixed size.","In the example discussed herein, a layer N PDU consisting of the actual signaling data 22 and the Integrity Check Info is discussed.","The Integrity Check Info consists of the 32 bit MAC-I 26 and the message sequence number SN 24, which is needed at the peer end for the recalculation of MAC-I.","The total length of the message 200 is then a combination of the signaling bits and the Integrity Check Info bits.","At the receiving end the message is received including the signaling data part and the Integrity Check Info (which comprises the message sequence number SN and the 32-bit MAC-I).","The signaling data together with the Integrity Check Info (ie the secret integrity the COUNT-I value, the direction of transmission, and the random number (FRESH)), are processed in a computing block, for example a function like the UTRAN f9 function, with a fixed function.","A thus generated received computed message authentication code XMAC-I is then compared with the message authentication code MAC-I received in the transmitted message.","If the two codes match (ie XMAC-I matches MAC-I), the recipient can trust that the message is intact, and the recipient then accepts the message.","Otherwise, the message is discarded.","The frame dependent COUNT-I number is actually the sum of a locally generated and incremented frame number HFN (Hyper Frame Number), which is added to the message sequence number, for example RRC_SN, and included in the message.","The HFN is incremented each time SN reaches its maximum value (SN is normally very short, e.g.","4 bits).","As mentioned hereinabove the transmitted.block (layer N-1 PDU) normally has a fixed length.","However, it may be that the signaling data bits together with the Integrity Check Info require more space than that provided in one layer N-1 PDU payload.","One known way to deal with this problem is to segment the signaling message.","In segmentation a signaling message, which is too long to fit in a single layer N-1 block, is passed on to a lower layer, where it is split up into two blocks (two layer N-1 PDUs), each with an appropriate layer N-1 header.","Two blocks is just an example here, naturally a larger message may require even more than two blocks.","If the second layer N-1 PDU is not totally filled with the layer N data, padding bits are inserted.","At the receiving end before transferring to a higher layer, the two layer N-1 payloads are reassembled into one layer N PDU.","To a person skilled in the art, it is immediately obvious that the use of padding bits is a potential waste of resources.","TDMA systems, for example, have a limited radio block size, whereby a message including the full message authentication code does not necessarily fit into one radio block.","This leads to the difficulty that the message has either to be sent without the MAC-I or in one or more additional segments.","In addition, there are certain time critical messages, for example, handover messages, which must be sent in one radio block only.","Generally, segmentation is not desirable, because it wastes radio resources and slows down the signaling procedure unnecessarily.","One way to solve the above problem is to make the length of the message authentication code shorter than 32 bits.","It has been proposed that such a message should include a field that defines the length of the message authentication code, a two-bit identifier, for example.","This identifier allows certain discrete values: 8, 16, 24 and 32.This solution still has some problems.","First, the identifier always takes two extra bits from the length of the message.","Second, the discrete values are not flexible, and in some cases this can lead to the same problem as above, i.e.","segmentation is needed for certain messages.","A particularly advantageous technique for addressing the above stated problems is disclosed in Finnish patent application number FI20002453.This discloses a technique that allows the transmission of a message in a single lower layer data block even when the length of the message including the integrity check info exceeds the length of the lower layer data block.","In all current known solutions, each message is authenticated separately.","Each message contains a sequence number of the protocol it belongs to.","Hence a binding between different steps in the existing solutions is achieved through the sequence number.","The drawback to existing solutions is that at each step a 32-bit MAC is transmitted which takes a significant part of the signaling bandwidth.","It is therefore an object to the present invention to provide an improved technique for taking the integrity of messages.","SUMMARY OF THE INVENTION In accordance with the present invention there is provided a method of providing message authentication in a communication system comprising the steps of: transmitting a first message from a first device to a second device; transmitting a second message from the second device to the first device, the second message including a message authentication code determined using said first and second messages; transmitting a third message from the first device to the second device, the third messages including a message authentication code determined using the third message.","In order to minimize the risk of ‘replay attacks’, the sequence numbers should preferably be maintained in the protocol.","This is particularly important if the messages are short or otherwise small in number.","The method may further comprise the step of storing the first message in the first device.","The method may further comprise the steps of: responsive to receipt of the second message, determining an expected message authentication code using said first and second messages; and comparing the expected message authentication code to the received message authentication code.","The method may further comprise the step of discarding the second message if the expected message authentication code does not match the received message authentication code.","The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message; and comparing the expected message authentication code to the received message authentication code.","The method may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.","The third message may include a message authentication code determined using the third message and the second message.","The method may further comprise the step of storing the second message in the second device.","The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message and said second message; and comparing the expected message authentication code to the received message authentication code.","The method according to claim 9 may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.","The third message may include a message authentication code determined using the third message, the second message, and the first message.","The method may further comprise the step of storing said first message in the second device.","The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message, said second message, and said first message; and comparing the expected message authentication code to the received message authentication code.","The method may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.","The invention may thus advantageously reduce the bandwidth used for authentication.","In a multi-step protocol, the invention provides for the messages exchanged at different steps to be grouped together for message authentication code computations.","A message is saved by the sending party for subsequent verification at a later step.","The number of required message authentication code computations and transmissions may be reduced to two independently of the number of steps in the authentication procedure.","In three step signaling procedures, there is thus no need for a message authentication code to be computed in the first step.","The invention is further advantageously applicable to procedures containing more than three messages.","All communications that occur between two parties in various steps may be authenticated by the communicating parties in the last two steps of the protocol.","The first and second devices are preferably elements of a mobile communication system.","BRIEF DESCRIPTION OF THE DRAWINGS The invention will be described more closely with reference to the accompanying drawings, in which FIG.","1 depicts the computation of a message authentication code; FIG.","2 shows the contents of a message; FIG.","3 illustrates a first embodiment of the present invention; FIG.","4 illustrates a second embodiment of the present invention; FIG.","5 depicts the creation of a message according to a preferred implementation; FIG.","6 is a flow chart showing the creation of a message in the GERAN system, and FIG.","7 is a flow chart of actions at the receiving end.","DESCRIPTION OF PREFERRED EMBODIMENTS The invention is described herein by way of reference to particular non-limiting examples, and with particular reference to a GERAN system.","The GERAN is specified by the Third Generation Partnership Project (3GPP).","GERAN is an evolution of the GSM-system (Global System for Mobile Communication), the TDMA/136-system (Time Division Multiple Access System), and the EDGE-system.","GERAN has no integrity protection of its own.","Implementation of the same integrity algorithms used in UTRAN is suggested for a radio system using the GPRS/EDGE-radio connection network GERAN.","This leads to certain significant problems, especially the problem of message segmentation.","The present invention can be advantageously used for implementing an integrity algorithm in the GERAN system, and the present invention is described herein with reference to example implementations in such a system.","However, the invention is not limited in its application to such a system.","Radio interface protocols are needed to set up, reconfigure, and release Radio Bearer services.","The protocol layers above the physical layer are called the data link layer (layer 2) and the network layer (layer 3).","The control plane layer 2 contains two sub-layers: Medium Access Control (MAC) protocol and Radio Link Control (RLC) protocol.","Layer 3 consists of one protocol, called Radio Recourse Control (RRC), which belongs to the control plane.","The channels offered by the physical layer to the MAC layer are called logical channels.","It shall be appreciated that the term ‘logical channel’ can be used for other purposes in other systems.","For example, in the UTRAN the term logical channel referes to a channel offered by the MAC layer to higher layers.","All higher layer signaling (mobility management, call control, session management, etc.)","is encapsulated into RRC messages for transmission over the radio interface.","The following provides a description of integrity protection for a message to be sent over a radio link.","Referring to FIG.","3, a first embodiment of the present invention is described.","For the purpose of this illustrative example, it is assumed that messages are being exchanged between the mobile station 40 and a network 42.It is assumed that the mobile station 40 initiates the exchanged messages.","The mobile station 40 constructs a message for transmission in the normal way in accordance with standard procedures.","Prior to transmitting the message, as indicated by step 50, the mobile station 40 stores the initial message, labeled message_1, in its memory.","As indicated by arrow 44, message_1 is then transmitted from the mobile station 40 to network 42.In accordance with the present invention, the first message is sent without a message authentication code.","The network 42 receives the first message from the mobile station, and prepares a second message, message_2, for transmission back to the mobile station 40.In addition, in a step 52 the network 42 calculates a first message authentication code, MAC_I_1, using both the first message as received by the mobile station and the second message which it is to transmit to the mobile station.","Thus the message authentication code to be transmitted with the second message is based on the combination of both the first and second messages.","As indicated by arrow 46, the network 42 then transmits a second message, message_2, with the message authentication code MAC_I_1.Using the received second message, message_2, and the stored first message, message_1, the mobile station 40 in a step 54 calculates the expected message authentication code XMAC_I_1.If XMAC_I_1 is identical to MAC_I_1, then the mobile station 40 continues with message transmission.","Otherwise the received message message_2 is discarded.","In this embodiment of the invention, the mobile station 40 prepares to transmit a third message message_3 to the network 42.In the step 56 the mobile station 40 prepares a second message authentication code MAC_I_2 using the content of the third message message_3.As represented by arrow 58, the third message message_3 is transmitted to the network 42 together with the second message authentication code MAC_I_2.The network 42 then calculates the expected message authentication code XMAC_I_2 using the third message message_3 in step 58, and in the normal way compares this to the transmitted authentication code MAC_I_2.Whilst in FIG.","3 the three messages are identified as message_1, message_2 and message_3, in actual signaling procedures they may be named, for example, XXX_request, XXX_command, XXX_complete (or “confirm”).","Thus in accordance with the first embodiment of the invention, the message authentication code is left out of the first message.","The message authentication code for the second message is calculated over the first and second messages, although the first message is not returned to the sender.","The third message is integrity protected “normally”, with a message authentication code calculated over the third message itself.","This technique ensures that the procedure cannot be used illegally by an intruder, even if the first message is not integrity protected.","Referring to FIG.","4, the second embodiment of the present invention is now described.","Where appropriate, the same reference numerals as used in FIG.","3 are used to refer to identical steps or procedures.","The embodiment of the invention described with reference to FIG.","4 is suitable for signaling procedures normally having either two or three messages.","As in the embodiment described hereinabove with reference to FIG.","3, the mobile station 40 prepares a first message message_1 for transmission to the network 42, and prior to transmitting it, as represented by arrow 44, stores it in a memory as represented by step 50.In step 52, the network 42 calculates a first message authentication code MAC_I_1 using both the first message and the second message.","In an additional step 53, the network 42 saves the second message message_2 into its memory before transmission.","As in the embodiment of FIG.","3, the network 42 transmits the second message message_2 together with the first message authentication code MAC_I_1 as indicated by arrow 46 to the mobile station 40.In step 54, as in the embodiment of FIG.","3, the mobile station 40 calculates the expected message authentication code XMAC_I_1 using both the received second message and the stored first message.","If the expected message authentication code XMAC_I_1 is identical to the received message authentication code MAC_I_1, then the procedure is continued with.","In this embodiment of the present invention, the mobile station 40 then calculates in step 55 a second message authentication code MAC_I_2 using both the third message to be sent, message_3, and the received second message, message_2.The mobile station 40 then transmits, as indicated by arrow 48, the third message, message_3, together with the message authentication code MAC_I_2.The network 42 then calculates the expected second message authentication code XMAC_I_2 using both the stored second message message_2 and the received third message message_3 in step 59.Thus, in common with the first embodiment described hereinabove with reference to FIG.","3, the second embodiment as described with reference to FIG.","4 similarly does not include any message authentication code in the first transmitted message.","The difference of the second embodiment compared to the first embodiment is that the third message contains a message authentication code calculated over both the second and third messages, whereas in the first embodiment the message authentication code is calculated over only the third message.","The technique of FIG.","4 may be more advantageous in situations where the third message is added only for this purpose, ie it does not contain any actual information but is merely an acknowledgement message and is thus very short.","From a security viewpoint, the calculation of the integrity check sum over a longer message is beneficial.","The addition of a third message may be more preferable than segmentation of the first message, because the segmentation solution necessitates additional acknowledgement (on data link layer level).","In a third embodiment of the present invention, the message authentication code transmitted with the third message is calculated over all three messages, ie message_1, message_2 and message_3.The advantage of this embodiment is that each check sum protects the maximum amount of the data communicated in the procedure.","Thus, this variation excludes the possibility of an attack where a “man in the middle” replaces the first message with another one and modifies the message authentication code in the second message accordingly.","Such an attack could only be successful if the attacker is able to modify the check sum correctly.","There are two possible ways to do this: 1) the replay of an earlier message authentication code; or 2) pure guessing.","The first way is only possible if the counter number repeats, which means the same integrity key has been in use for too long.","The possibility of the second way being successful is a very low probability.","However, if the message authentication code of the second message is not a full 32-bits long, the probability of the guessing attack becomes higher.","If the “man in the middle” is to have any chance of making a successful attack when the third embodiment of the invention is in use, then it is necessary for either 1) or 2) to be succeeded twice, which is much more difficult.","The present invention can particularly advantageously be used in combination with the technique described in Finnish patent application number FI120002453.Such a technique is described hereinbelow.","It should be noted that the techniques described hereinbelow may only be utilized in steps where a message authentication code is being generated, and hence would not be used in accordance with the invention in relation to the generation and transmission of the first message.","It should also be noted that other techniques may also be used.","FIG.","5 illustrates a situation where a signaling message 500 is to be sent in a secure manner over a lower layer radio link in one fixed length radio block, which can be a TDMA block, for example, without segmentation.","The maximum block size allowed by the lower layer data block 501 is indicated by dotted lines in the figure.","The signaling message without the Integrity Check Info (ICI) is in the illustrated situation shorter than the said maximum block size.","This leads to a situation where the data to be sent either has to be segmented or sent without the message authentication code.","Neither of these alternatives is acceptable.","In order for the data to be sent in a sufficiently secure manner over a radio link, the computed message authentication code should be appended to it.","However, it must be shortened in a predefined way (described in detail below).","This truncated message authentication code diminishes the reliability of the integrity protection to some degree but still provides sufficient protection for the message.","It should be noted, that the sequential number SN needed to form the COUNT-I parameter cannot be truncated.","The message authentication code may be computed in the usual way in the device concerned and the MAC-I added with the message sequence number to the encoded message to form the actual PDU.","Then the length of the message (without the Integrity Check Info) and/or the length of the PDU can be examined as follows.","i) If the length of the message is longer than the length of the lower layer data block, the PDU is segmented into two or more data blocks as in prior art.","ii) If the length of the PDU is shorter than the length of the lower layer data block, the PDU is placed into said lower layer data block and the rest of the block is filled with padding bits (normally by the lower layer itself).","iii) If the length of the PDU is longer than the length of the lower layer data block but the extra bits are less than the size of the MAC-I, then the computed message authentication code is truncated so that the truncated PDU fits into one layer N-1 data block.","However, truncation of the message authentication code diminishes the security of the message exchange.","Therefore, the number of bits the MAC-I is allowed to be truncated by is limited to a certain maximum value, i.e.","the truncated message authentication code has a certain minimum value.","Thereafter, the truncated PDU is sent via a radio interface to the receiving end.","At the receiving end the integrity is examined of the PDU received.","First, the part including the signaling bits and the part including the Integrity Check Info are separated.","Then a message authentication code is recomputed based on exactly the same algorithm and using the same parameters as were used at the transmitting end.","The message authentication code of the received message is then compared with the recomputed authentication code.","FIG.","6 shows as a flowchart a more detailed example of one implementation of the method according to the invention from the point of view of the transmitting end.","At stage 600 a time critical RRC message is to be sent through a radio interface, for example from the network to a mobile.","Most signaling messages sent between a mobile station MS and the network, for example, must be integrity protected.","Examples of such messages are RRC, MM, CC, GMM, and SM messages.","Integrity protection is applied at the RRC layer, both in the mobile station and in the network.","Integrity protection is usually performed for all RRC (Radio Recourse Control) messages, with some exceptions.","These exceptions can be: 1.messages that are assigned to more than one recipient, —messages that have been sent before integrity keys were created for the connection, 2.frequently repeated messages, including information which does not need integrity protection.","The message is encoded according to the specified message transfer syntax at stage 601.The encoded message (bit string) is called here E. A 32-bit message authentication code MAC-I, which is to be added to the encoded message, is calculated at stage 602.The message authentication code not only depend on the encoded message but also on several other parameters.","The following input parameters are needed for calculation of the integrity algorithm: the encoded message, the 4-bit sequence number SN, the 28-bit hyper-frame number HFN, the 32-bit random number FRESH, the 1-bit direction identifier DIR, and the most important parameter—the128-bit integrity key IK.","The short sequence number SN and the long sequence number HFN together compose the serial integrity sequence number COUNT-I.","When the message authentication code is computed using the UMTS integrity algorithm and the above parameters, it is guaranteed that no one other than the actual sender can add the correct MAC-I code to the signaling message.","COUNT-I, for example, prevents the same message from being sent repeatedly.","However, if the same signaling message for some reason or other is to be sent repeatedly, the MAC-I code differs from the MAC-I code that was in the previously sent signaling message.","The aim of that is to protect the message as strongly as possible against eavesdroppers and other fraudulent users.","Due to the fact that a TDMA radio block has a fixed length, the length of the message has to be checked to avoid segmentation of the message.","The RRC layer makes a decision as to whether the segmentation of the message concerned is to be allowed or not.","At stage 603 the total length of the signaling message to be sent without the message authentication code is calculated using the following formula: X=max_size−sizeof(E)−sizeof(RRC_SN) In the above formula max_size is the maximum size (in bits) of a RRC message that can be sent in one radio block (i.e.","there is no need for segmentation).","Sizeof(E) is the size (in bits) of the encoded message and sizeof(RRC_SN) is the size of the RRC sequence number, a 4-bit working assumption.","X defines the length (in bits) of the rest of the fixed length message, which is still left after the minimum number of bits are reserved for the message authentication code, the untruncated MAC-I size may be different.","Next, at stage 604, a comparison is made to ascertain whether the calculated X is between values 0 and min_MACI_len, where the latter value is the minimum allowed length for the message authentication code.","This minimum length is a predefined value, which can be either the same for all messages or even a message type specific value.","It is clear that the smaller the value, the weaker the protection.","So it is obvious that a minimum length must be determined so that the message can be sent with adequate security.","If the answer after said comparison is YES, this means that the message authentication code does not fit with the signaling message to be sent in one radio block.","In other words, the space left in one radio block is too short even for a shortened message authentication code after the signaling message is put into the block.","If this is the case, the system protocol defines 605 as the next action to be carried out.","If the answer after comparison is NO, the next step is to compare whether X is between values min_MACI_len and 32, stage 606.If X is between those values, i.e.","if the answer after the comparison in stage 604 is YES, then X bits of the message authentication code with the RRC_SN are added to the encoded message, stage 607.The sequence number RRC_SN is needed for integrity protection, that is, for calculation of the message authentication code at the receiving end.","Note that the MAC-I size is also 32 bits in the UTRAN system.","In some other systems, the ‘normal’ MAC-I size may be something different.","The length of the message authentication code is shortened in a predefined way.","Thus, the size of the message authentication code transmitted over the radio path depends dynamically on the size of each encoded message, not on the type of the message.","The decisions are made at the RRC layer as to the minimum message authentication code size and as to when the size of the said code may be shortened.","In some cases the RRC layer can make the decision that the message authentication code is not to be shortened even though this would have been possible.","Such cases might occur when strong protection is demanded for a message.","The next step 611 is to send the message including the integrity protection info (E+MAC-I+RRC_SN) to the lower layers for transmission over the radio interface to the mobile station.","If the answer in the above comparison 604 is NO, a final comparison is made as to whether the value of X is greater than 31, stage 608.If the answer is YES, this means that neither shortening nor segmentation is needed.","Now the whole message authentication code and the RRC_SN are appended to the encoded message E, stage 609.The next step is stage 611.If the answer to the comparison at stage 608 is NO, i.e.","if X is smaller than 0, which means that the size of the encoded message sizeof(E) is greater than the maximum size of the RRC message max_size, then two different alternatives A and B, are possible at stage 610: 1.add the entire MAC-I (+RRC_SN) to the message; 2.set sizeof(E)=(sizeof(E)−max_size) and rerun the previous steps.","Which of the two above alternatives is selected depends on the protocol according to the system.","Alternative A means that the whole message authentication code with the sequence number RRC_SN is added to the block, since the message has to be segmented anyway.","Thus with this alternative, it is not important whether adding Integrity Protection Info causes additional segmentation or not.","In alternative B, the attempt is made to avoid the additional segmentation caused by the addition of Integrity Check Info.","Thus the sizeof(E) is set one full data block shorter than what was given in stage 601, and the truncation algorithm for the MAC-I is rerun starting from step 603.FIG.","7 shows as a flowchart an example of one implementation of the method according to the invention from the point of view of the receiving end.","At stage 700 the receiving end gets a Service Data Unit (SDU) comprising the signaling data M from the lower layers.","It is assumed here that this message is the same as in the previous example in FIG.","6.The next step is that the part including signaling data bits and the part including the Integrity Check Info MAC-I (the message authentication code with the sequence number the RRC_SN) are separated and a message authentication code with RRC sequence number RRC_SN is decoded in stage 701.The actual message (signaling data) can still be stored as an encoded bit string at this point.","In prior art systems the message received is discarded immediately if the received and the recomputed message authentication codes do not match.","But according to the described embodiment of the invention, the receiving end first examines the length of the message authentication code and, depending on the result, it then decides, if and how the message is to be processed further.","For example, if the receiving end finds that the message authentication code received is shorter than it should be, it may assume that the code has been truncated.","Instead of n bits the truncated code comprises m bits.","If the truncation exceeds the predetermined maximum amount known by the receiving end, the message is discarded.","If truncation does not exceed the predetermined maximum amount as known by the receiving end, the bits of the truncated message authentication code are compared bit-by-bit to the bits of the recomputed authentication code of full length.","When the m bits of the truncated message authentication code match the corresponding bits of the recomputed message authentication code, the integrity check of the message received is passed.","The length of the message authentication code of the message received is examined at stages 702 and 706.In addition, the length of the entire message (including the signaling data and the integrity check info) received is also examined at stage 704.At stage 702 a check is made as to whether the length of the MAC-I is ‘normal’, in this example 32 bits.","If YES (the answer to stage 702 is NO), the flow proceeds to stage 703, where the message is processed in the normal way.","The message authentication code is checked, the message is decoded, etc.","using the same algorithm and parameters as were used at the transmitting end.","Provided that stage 702 yields the YES alternative, meaning that the MAC-I has been truncated, a check is made as to whether the length of the message is a multiple of the max_size (sizeof(M) mod max_size—0), justifying the MAC-I truncation.","A NO alternative yields a protocol error, for which reason the message received is discarded 705.A YES answer after stage 704 leads to stage 706, when a comparison is made as to whether the length of the message authentication code is greater than or equal to the minimum allowed MAC-I length (min_MACI_len).","A NO alternative yields a protocol error, for which reason the received message is discarded 707.If the length is greater than or equal to the minimum value, the transmitting end may have shortened the MAC-I code in the correct way.","With the integrity key and all the other needed parameters, the expected message authentication code XMAC-I is calculated 708, using the same algorithm as for the transmitting end.","The calculated XMAC-I has to be truncated in order to compare its size with the size of the transmitted MAC-I, stage 709.If the truncated XMAC-I does not correspond to the transmitted truncated MAC-I 710, an integrity error is found and the received message is discarded 711.If the result of the comparison is positive, the message is decoded 712.The final check 713 is made after decoding the actual signaling data 712.The final check is to find out whether there are some padding bits in the received message.","Since the MAC-I has been truncated due to message size, no padding bits are allowed (since the padding bits should have been used for the MAC-I).","The Integrity check is OK whenever no RRC padding bits are found 713, i.e.","it is then ensured that the message has been sent from the authorized party 715.Otherwise, a protocol error is found and the message is discarded, stage 714.The above defined preferred technique for generation of the message authentication code may be used in the embodiments of the present invention at those steps where a message authentication code is generated.","An implementation and embodiment of the present invention has been explained above with some examples.","However, it is understood that the invention is not restricted to the details of the above embodiment and that numerous changes and modifications can be made by those skilled in the art without departing from the characteristic features of the invention.","The embodiment described is to be considered illustrative but not restrictive.","Therefore, the invention should be limited only by the attached claims.","Thus, alternative implementations defined by the claims, as well as equivalent implementations, are included in the scope of the invention.","For example, instead of at the RRC layer the decision concerning the MAC-I size can be made at some other layer, e.g.","the RLC layer.","In that case, the RLC must know whether segmentation of the message(s) in the transmission buffer is to be allowed or not.","The protocol layers from top to bottom may be, for example, RRC, LLC (Logical Link Control), LAPDm (Link Access Protocol on the Dm channel), PDCP (Packet Data Convergence Protocol), RLC, MAC (Medium Access Control Protocol), and PHY (Physical Layer) In addition, the minimum value, which is set at min_MACI_len might also depend on the signaling message used.","The grouping of signaling messages into different min_MACI_len categories can be carried out either simply according to the message type.","Grouping can be based on other factors as well, such as on whether the message is so that for critical signaling messages the value is greater than for non-critical signaling messages.","For some non-critical messages the min_MACI_len could be set as low as 8 bits, for example.","Utilised in combination with the techniques described with reference to FIGS.","5 to 7 the present invention may allow, in certain situations, a longer MAC-I, thereby further improving integrity.","The present invention is independent of the length of the message authentication code.","Such code may be longer or shorter than 32 bits.","The length may also differ between messages in one procedure.","It should also be noted that although this application is made only from the signaling standpoint, integrity protection can also be applied in some systems to the user plane data.","The same principles and methods described in this application are applicable also for user plane data packets, although the actual protocol layers performing the integrity protection, and the message authentication code truncation would then be different."]],"string":"[\n [\n \"Message authentication\",\n \"There is disclosed a technique of providing message authentication in a communication system comprising the steps of: transmitting a first message from a first device to a second device; transmitting a second message from the second device to the first device, the second message including a message authentication code determined using said first and second messages; transmitting a third message from the first device to the second device, the third messages including a message authentication code determined using the third message.\",\n \"The message authentication code of the third message may be additionally based on the second or the second and first messages.\"\n ],\n [\n \"1.A method of providing message authentication in a communication system comprising the steps of: transmitting a first message from a first device to a second device; transmitting a second message from the second device to the first device, the second message including a message authentication code determined using said first and second messages; transmitting a third message from the first device to the second device, the third message including a message authentication code determined using the third message.\",\n \"2.A method according to claim 1, further comprising the step of storing the first message in the first device.\",\n \"3.A method according to claim 1 further comprising the steps of: responsive to receipt of the second message, determining an expected message authentication code using said first and second messages; and comparing the expected message authentication code to the received message authentication code.\",\n \"4.A method according to claim 3 further comprising the step of discarding the second message if the expected message authentication code does not match the received message authentication code.\",\n \"5.A method according to claim 1 further comprising the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message; and comparing the expected message authentication code to the received message authentication code.\",\n \"6.A method according to claim 5 further comprising the step of discarding the third message if the expected message authentication code does not match the received message authentication code.\",\n \"7.A method according to claim 1, wherein the third message includes a message authentication code determined using the third message and the second message.\",\n \"8.A method according to claim 7, further comprising the step of storing the second message in the second drive.\",\n \"9.A method according to claim 7, further comprising the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message and said second message; and comparing the expected message authentication code to the received message authentication code.\",\n \"10.A method according to claim 9 further comprising the step of discarding the third message if the expected message authentication code does not match the received message authentication code.\",\n \"11.A method according to claim 1, wherein the third message includes a message authentication determined using the third message, the second message, and the first message.\",\n \"12.A method according to claim 11, further comprising the step of storing said first message in the second device.\",\n \"13.A method according to claim 11, further comprising the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message, said second message, and said first message; and comparing the expected message authentication code to the received message authentication code.\",\n \"14.A method according to claim 13 further comprising the step of discarding the third message if the expected message authentication code does not match the received message authentication code.\",\n \"15.A method according to claim 1 wherein the first and second devices are elements of a mobile communication system.\",\n \"16.A method of providing message authentication in a communication system comprising transmitting a plurality of messages between a first device and a second device, wherein a message includes a message authentication code determined using at least said message and another message includes a further message authentication code determined using at least said other message, whereby the number of required message authentication code computations and transmissions may be reduced to two independently of the number of steps of the authentication procedure.\",\n \"17.A communication system for providing message authentication between two communicating devices, each communicating device having respective transmitting and receiving means, and each communicating device having means for generating a message authentication code, comprising: in the first device, transmitting a first message from the second device; in the second device, transmitting a second message to the first device, the second message including a message authentication code determined using said first and second messages; in the first device transmitting a third message to the second device, the third messages including a message authentication code determined using the third message.\",\n \"18.A communication system according to claim 17, further including, in the first device: responsive to receipt of the second message, determining an expected message authentication code using said first and second messages; and comparing the expected message authentication code to the received message authentication code.\",\n \"19.A communication system according to claim 17 wherein the second message is discarded if the expected message authentication code does not match the received message authentication code.\",\n \"20.A communication system according to claim 17 further comprising, in the second device: responsive to receipt of the third message, determining an expected message authentication code using said third message; and comparing the expected message authentication code to the received message authentication code.\",\n \"21.A communication system according to claim 20 wherein the third message is discarded if the expected message authentication code does not match the received message authentication code.\",\n \"22.A communication system according to claim 17, wherein the first device generates the third message including a message authentication code determined using the third message and the second message.\",\n \"23.A communications system according to claim 22, wherein responsive to receipt of the third message, the second device determines an expected message authentication code using said third message and said second message; and compares the expected message authentication code to the received message authentication code.\",\n \"24.A communications system according to claim 23 wherein the third message if the expected message authentication code does not match the received message authentication code.\",\n \"25.A communications system according to claim 17 wherein the third message generated by the first device includes a message authentication code determined using the third message, the second message, and the first message.\",\n \"26.A communication system according to claim 25 wherein responsive to receipt of the third message, the second device determines an expected message authentication code using said third message, said second message, and said first message; and compares the expected message authentication code to the received message authentication code.\",\n \"27.A communication system according to claim 26 wherein the second device discards the third message if the expected message authentication code does not match the received message authentication code.\",\n \"28.A communication system according to claim 17 wherein the first and second device are elements of a mobile communication system.\",\n \"29.A communication system according to claim 28, wherein the first and second devices are one of a mobile terminal and a network element.\",\n \"30.A communication system according to claim 28, wherein the mobile communication system comprises a GERAN system.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>All telecommunication is subject to the problem of how to make sure that the information received has been sent by an authorized sender and not by somebody who is trying to masquerade as the sender.\",\n \"The problem is particularly evident in cellular telecommunication systems, where the air interface presents a potential platform for eavesdropping and replacing the contents of a transmission by using higher transmission levels, even from a distance.\",\n \"A basic solution to this problem is the authentication of the communicating parties.\",\n \"An authentication process aims to discover and check the identity of both the communicating parties, so that each party receives information about the identity of the other party and can rely on the identification to a sufficient degree.\",\n \"Authentication is typically performed in a specific procedure at the beginning of a connection.\",\n \"However, this does not adequately protect subsequent messages from unauthorized manipulation, insertion, and deletion.\",\n \"Thus, there is a need for the separate authentication of each transmitted message.\",\n \"The latter task can be carried out by appending a message authentication code (MAC-I) with a particular value to the message at the transmitting end and checking the MAC-I value at the receiving end.\",\n \"A MAC-I is typically a relatively short string of bits based in some specified way on the message it protects and on a secret key known both by the sender and by the recipient of the message.\",\n \"The secret key is generated and agreed on typically in connection with the authentication procedure at the beginning of the connection.\",\n \"In some cases the algorithm that is used to calculate the MAC-I based on the secret key and on the message may also be secret.\",\n \"The process of authentication of single messages is often called integrity protection.\",\n \"To protect the integrity of signaling, the transmitting party computes a MAC-I value based on the message to be sent and the secret key using the specified algorithm, and sends the message with the MAC-I value.\",\n \"The receiving party recomputes a MAC-I value based on the message and the same secret key according to the same specified algorithm, and compares the received MAC-I and the calculated MAC-I.\",\n \"If the two MAC-I values match, the recipient can trust that the message is intact and has been sent by the authorized party.\",\n \"Known integrity protection schemes are not completely reliable.\",\n \"A third party can attempt to manipulate a message transmitted between a first and a second party.\",\n \"There are two main alternative methods for forging a MAC-I value for a modified or a new message: obtaining the secret key; or by trying directly without the secret key.\",\n \"The secret key can be obtained by a third party in two ways: i) by computing all possible keys until a key is found matching the data of observed message MAC-I pairs, or by otherwise breaking the algorithm for producing MAC-I values; or ii) by directly capturing a stored or transmitted secret key.\",\n \"The original communicating parties can prevent a third party from obtaining the secret key by using an algorithm which is cryptographically strong and which uses a secret key which is long enough to prevent the exhaustive search of all keys, and by using other security means for the transmission and storage of secret keys.\",\n \"A third party may try to disrupt the sending of messages between the two parties without a secret key by guessing the correct MAC-I value, or by replaying some earlier message transmitted between the two parties for which message the correct MAC-I is known from the original transmission.\",\n \"Guessing of the correct MAC-I value can be made difficult by using long MAC-I values.\",\n \"The MAC-I value should be long enough to reduce the probability of correct guessing to a sufficiently low level compared to the benefit gained by one successful forgery.\",\n \"For example, using a 32 bit MAC-I value reduces the probability of a correct guess to 1/4294967296, which is small enough for most applications.\",\n \"Obtaining a correct MAC-I value using the replay attack, i.e.\",\n \"by re-playing an earlier message, can be prevented by introducing a varying parameter to the calculation of the MAC-I values.\",\n \"For example, a time stamp value, a sequence number, or a random number can be used as further input to the MAC-I algorithm, in addition to the secret integrity key and the message.\",\n \"In the following, the prior art methods are described in more detail.\",\n \"When using sequence numbers, each party has to keep track of which sequence numbers have already been used and are not acceptable any more.\",\n \"The easiest way to implement this is to store the highest sequence number used in MAC-I calculations so far.\",\n \"This approach has the drawback that between connections each party must maintain state information which is at least to some level synchronized.\",\n \"That is, they need to store the highest sequence number used so far.\",\n \"This requires the use of a large database in the network.\",\n \"A further approach is to include a random number in each message, which the other side must use in MAC-I calculation the next time a message is sent for which MAC-I authentication is required.\",\n \"This approach has the same drawback as the previous one, i.e.\",\n \"between connections each party must maintain state information, which requires the use of a large database in the network.\",\n \"By way of example, FIG.\",\n \"1 illustrates the computation of a message authentication code in the UTRAN (UMTS Terrestrial Radio Access Network), which is a wideband multiple access radio network currently being specified in the 3GPP (Third Generation Partnership Project).\",\n \"The length of the MAC-I used in UTRAN is 32 bits.\",\n \"Block 100 represents the UMTS integrity algorithm for generating the message authentication code.\",\n \"The UMTS integrity algorithm used in block 100 is a one-way cryptographic function for calculating the 32 bit Message Authentication Code (MAC-I) based on the five input parameters shown in FIG.\",\n \"1 .\",\n \"A one-way function makes it impossible to derive the unknown input parameters from a MAC-I, even if all but one input parameter are known.\",\n \"The input parameters for calculating the MAC-I 20 are: the actual signaling message 10 (after encoding) to be sent, a secret integrity key 12 , a number COUNT-I value 14 for the message to be integrity protected, a value indicating the direction of transmission 16 , (i.e.\",\n \"whether the message is sent in uplink or downlink direction), and a random number 18 (FRESH) generated by the network.\",\n \"The COUNT-I value consists of a hyper frame number HFN and the message sequence number SN.\",\n \"The computing block 100 calculates the message authentication code by applying the afore-mentioned parameters to the integrity algorithm, which is called the f9 algorithm in 3GPP Release'99 specifications.\",\n \"FIG.\",\n \"2 illustrates a typical message to be sent over a radio interface.\",\n \"The message is a layer N protocol data unit (PDU) 200 , which is transferred as a payload 30 in a layer N- 1 PDU 201 .\",\n \"In the present example, layer N represents the Radio Resource Control (RRC) protocol in the radio interface and layer N- 1 represents the Radio Link Control (RLC) layer.\",\n \"The layer N- 1 PDU normally has a fixed size, which depends on the physical layer (the lowest layer, not visible in FIG.\",\n \"2 ) the channel type used and on other parameters, e.g.\",\n \"modulation, channel coding, interleaving.\",\n \"If layer N PDUs are not exactly the size of the payload 30 offered by layer N- 1 as is normally the case, layer N- 1 can utilize functions like segmentation, concatenation, and padding to make layer N- 1 PDUs a fixed size.\",\n \"In the example discussed herein, a layer N PDU consisting of the actual signaling data 22 and the Integrity Check Info is discussed.\",\n \"The Integrity Check Info consists of the 32 bit MAC-I 26 and the message sequence number SN 24 , which is needed at the peer end for the recalculation of MAC-I.\",\n \"The total length of the message 200 is then a combination of the signaling bits and the Integrity Check Info bits.\",\n \"At the receiving end the message is received including the signaling data part and the Integrity Check Info (which comprises the message sequence number SN and the 32-bit MAC-I).\",\n \"The signaling data together with the Integrity Check Info (ie the secret integrity the COUNT-I value, the direction of transmission, and the random number (FRESH)), are processed in a computing block, for example a function like the UTRAN f9 function, with a fixed function.\",\n \"A thus generated received computed message authentication code XMAC-I is then compared with the message authentication code MAC-I received in the transmitted message.\",\n \"If the two codes match (ie XMAC-I matches MAC-I), the recipient can trust that the message is intact, and the recipient then accepts the message.\",\n \"Otherwise, the message is discarded.\",\n \"The frame dependent COUNT-I number is actually the sum of a locally generated and incremented frame number HFN (Hyper Frame Number), which is added to the message sequence number, for example RRC_SN, and included in the message.\",\n \"The HFN is incremented each time SN reaches its maximum value (SN is normally very short, e.g.\",\n \"4 bits).\",\n \"As mentioned hereinabove the transmitted.block (layer N- 1 PDU) normally has a fixed length.\",\n \"However, it may be that the signaling data bits together with the Integrity Check Info require more space than that provided in one layer N- 1 PDU payload.\",\n \"One known way to deal with this problem is to segment the signaling message.\",\n \"In segmentation a signaling message, which is too long to fit in a single layer N- 1 block, is passed on to a lower layer, where it is split up into two blocks (two layer N- 1 PDUs), each with an appropriate layer N- 1 header.\",\n \"Two blocks is just an example here, naturally a larger message may require even more than two blocks.\",\n \"If the second layer N- 1 PDU is not totally filled with the layer N data, padding bits are inserted.\",\n \"At the receiving end before transferring to a higher layer, the two layer N- 1 payloads are reassembled into one layer N PDU.\",\n \"To a person skilled in the art, it is immediately obvious that the use of padding bits is a potential waste of resources.\",\n \"TDMA systems, for example, have a limited radio block size, whereby a message including the full message authentication code does not necessarily fit into one radio block.\",\n \"This leads to the difficulty that the message has either to be sent without the MAC-I or in one or more additional segments.\",\n \"In addition, there are certain time critical messages, for example, handover messages, which must be sent in one radio block only.\",\n \"Generally, segmentation is not desirable, because it wastes radio resources and slows down the signaling procedure unnecessarily.\",\n \"One way to solve the above problem is to make the length of the message authentication code shorter than 32 bits.\",\n \"It has been proposed that such a message should include a field that defines the length of the message authentication code, a two-bit identifier, for example.\",\n \"This identifier allows certain discrete values: 8, 16, 24 and 32.This solution still has some problems.\",\n \"First, the identifier always takes two extra bits from the length of the message.\",\n \"Second, the discrete values are not flexible, and in some cases this can lead to the same problem as above, i.e.\",\n \"segmentation is needed for certain messages.\",\n \"A particularly advantageous technique for addressing the above stated problems is disclosed in Finnish patent application number FI20002453.This discloses a technique that allows the transmission of a message in a single lower layer data block even when the length of the message including the integrity check info exceeds the length of the lower layer data block.\",\n \"In all current known solutions, each message is authenticated separately.\",\n \"Each message contains a sequence number of the protocol it belongs to.\",\n \"Hence a binding between different steps in the existing solutions is achieved through the sequence number.\",\n \"The drawback to existing solutions is that at each step a 32-bit MAC is transmitted which takes a significant part of the signaling bandwidth.\",\n \"It is therefore an object to the present invention to provide an improved technique for taking the integrity of messages.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>In accordance with the present invention there is provided a method of providing message authentication in a communication system comprising the steps of: transmitting a first message from a first device to a second device; transmitting a second message from the second device to the first device, the second message including a message authentication code determined using said first and second messages; transmitting a third message from the first device to the second device, the third messages including a message authentication code determined using the third message.\",\n \"In order to minimize the risk of ‘replay attacks’, the sequence numbers should preferably be maintained in the protocol.\",\n \"This is particularly important if the messages are short or otherwise small in number.\",\n \"The method may further comprise the step of storing the first message in the first device.\",\n \"The method may further comprise the steps of: responsive to receipt of the second message, determining an expected message authentication code using said first and second messages; and comparing the expected message authentication code to the received message authentication code.\",\n \"The method may further comprise the step of discarding the second message if the expected message authentication code does not match the received message authentication code.\",\n \"The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message; and comparing the expected message authentication code to the received message authentication code.\",\n \"The method may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.\",\n \"The third message may include a message authentication code determined using the third message and the second message.\",\n \"The method may further comprise the step of storing the second message in the second device.\",\n \"The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message and said second message; and comparing the expected message authentication code to the received message authentication code.\",\n \"The method according to claim 9 may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.\",\n \"The third message may include a message authentication code determined using the third message, the second message, and the first message.\",\n \"The method may further comprise the step of storing said first message in the second device.\",\n \"The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message, said second message, and said first message; and comparing the expected message authentication code to the received message authentication code.\",\n \"The method may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.\",\n \"The invention may thus advantageously reduce the bandwidth used for authentication.\",\n \"In a multi-step protocol, the invention provides for the messages exchanged at different steps to be grouped together for message authentication code computations.\",\n \"A message is saved by the sending party for subsequent verification at a later step.\",\n \"The number of required message authentication code computations and transmissions may be reduced to two independently of the number of steps in the authentication procedure.\",\n \"In three step signaling procedures, there is thus no need for a message authentication code to be computed in the first step.\",\n \"The invention is further advantageously applicable to procedures containing more than three messages.\",\n \"All communications that occur between two parties in various steps may be authenticated by the communicating parties in the last two steps of the protocol.\",\n \"The first and second devices are preferably elements of a mobile communication system.\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention generally relates to a method for checking the integrity of messages in a communication system, particularly but not exclusively between a mobile station and a cellular network.\",\n \"BACKGROUND OF THE INVENTION All telecommunication is subject to the problem of how to make sure that the information received has been sent by an authorized sender and not by somebody who is trying to masquerade as the sender.\",\n \"The problem is particularly evident in cellular telecommunication systems, where the air interface presents a potential platform for eavesdropping and replacing the contents of a transmission by using higher transmission levels, even from a distance.\",\n \"A basic solution to this problem is the authentication of the communicating parties.\",\n \"An authentication process aims to discover and check the identity of both the communicating parties, so that each party receives information about the identity of the other party and can rely on the identification to a sufficient degree.\",\n \"Authentication is typically performed in a specific procedure at the beginning of a connection.\",\n \"However, this does not adequately protect subsequent messages from unauthorized manipulation, insertion, and deletion.\",\n \"Thus, there is a need for the separate authentication of each transmitted message.\",\n \"The latter task can be carried out by appending a message authentication code (MAC-I) with a particular value to the message at the transmitting end and checking the MAC-I value at the receiving end.\",\n \"A MAC-I is typically a relatively short string of bits based in some specified way on the message it protects and on a secret key known both by the sender and by the recipient of the message.\",\n \"The secret key is generated and agreed on typically in connection with the authentication procedure at the beginning of the connection.\",\n \"In some cases the algorithm that is used to calculate the MAC-I based on the secret key and on the message may also be secret.\",\n \"The process of authentication of single messages is often called integrity protection.\",\n \"To protect the integrity of signaling, the transmitting party computes a MAC-I value based on the message to be sent and the secret key using the specified algorithm, and sends the message with the MAC-I value.\",\n \"The receiving party recomputes a MAC-I value based on the message and the same secret key according to the same specified algorithm, and compares the received MAC-I and the calculated MAC-I.\",\n \"If the two MAC-I values match, the recipient can trust that the message is intact and has been sent by the authorized party.\",\n \"Known integrity protection schemes are not completely reliable.\",\n \"A third party can attempt to manipulate a message transmitted between a first and a second party.\",\n \"There are two main alternative methods for forging a MAC-I value for a modified or a new message: obtaining the secret key; or by trying directly without the secret key.\",\n \"The secret key can be obtained by a third party in two ways: i) by computing all possible keys until a key is found matching the data of observed message MAC-I pairs, or by otherwise breaking the algorithm for producing MAC-I values; or ii) by directly capturing a stored or transmitted secret key.\",\n \"The original communicating parties can prevent a third party from obtaining the secret key by using an algorithm which is cryptographically strong and which uses a secret key which is long enough to prevent the exhaustive search of all keys, and by using other security means for the transmission and storage of secret keys.\",\n \"A third party may try to disrupt the sending of messages between the two parties without a secret key by guessing the correct MAC-I value, or by replaying some earlier message transmitted between the two parties for which message the correct MAC-I is known from the original transmission.\",\n \"Guessing of the correct MAC-I value can be made difficult by using long MAC-I values.\",\n \"The MAC-I value should be long enough to reduce the probability of correct guessing to a sufficiently low level compared to the benefit gained by one successful forgery.\",\n \"For example, using a 32 bit MAC-I value reduces the probability of a correct guess to 1/4294967296, which is small enough for most applications.\",\n \"Obtaining a correct MAC-I value using the replay attack, i.e.\",\n \"by re-playing an earlier message, can be prevented by introducing a varying parameter to the calculation of the MAC-I values.\",\n \"For example, a time stamp value, a sequence number, or a random number can be used as further input to the MAC-I algorithm, in addition to the secret integrity key and the message.\",\n \"In the following, the prior art methods are described in more detail.\",\n \"When using sequence numbers, each party has to keep track of which sequence numbers have already been used and are not acceptable any more.\",\n \"The easiest way to implement this is to store the highest sequence number used in MAC-I calculations so far.\",\n \"This approach has the drawback that between connections each party must maintain state information which is at least to some level synchronized.\",\n \"That is, they need to store the highest sequence number used so far.\",\n \"This requires the use of a large database in the network.\",\n \"A further approach is to include a random number in each message, which the other side must use in MAC-I calculation the next time a message is sent for which MAC-I authentication is required.\",\n \"This approach has the same drawback as the previous one, i.e.\",\n \"between connections each party must maintain state information, which requires the use of a large database in the network.\",\n \"By way of example, FIG.\",\n \"1 illustrates the computation of a message authentication code in the UTRAN (UMTS Terrestrial Radio Access Network), which is a wideband multiple access radio network currently being specified in the 3GPP (Third Generation Partnership Project).\",\n \"The length of the MAC-I used in UTRAN is 32 bits.\",\n \"Block 100 represents the UMTS integrity algorithm for generating the message authentication code.\",\n \"The UMTS integrity algorithm used in block 100 is a one-way cryptographic function for calculating the 32 bit Message Authentication Code (MAC-I) based on the five input parameters shown in FIG.\",\n \"1.A one-way function makes it impossible to derive the unknown input parameters from a MAC-I, even if all but one input parameter are known.\",\n \"The input parameters for calculating the MAC-I 20 are: the actual signaling message 10 (after encoding) to be sent, a secret integrity key 12, a number COUNT-I value 14 for the message to be integrity protected, a value indicating the direction of transmission 16, (i.e.\",\n \"whether the message is sent in uplink or downlink direction), and a random number 18 (FRESH) generated by the network.\",\n \"The COUNT-I value consists of a hyper frame number HFN and the message sequence number SN.\",\n \"The computing block 100 calculates the message authentication code by applying the afore-mentioned parameters to the integrity algorithm, which is called the f9 algorithm in 3GPP Release'99 specifications.\",\n \"FIG.\",\n \"2 illustrates a typical message to be sent over a radio interface.\",\n \"The message is a layer N protocol data unit (PDU) 200, which is transferred as a payload 30 in a layer N-1 PDU 201.In the present example, layer N represents the Radio Resource Control (RRC) protocol in the radio interface and layer N-1 represents the Radio Link Control (RLC) layer.\",\n \"The layer N-1 PDU normally has a fixed size, which depends on the physical layer (the lowest layer, not visible in FIG.\",\n \"2) the channel type used and on other parameters, e.g.\",\n \"modulation, channel coding, interleaving.\",\n \"If layer N PDUs are not exactly the size of the payload 30 offered by layer N-1 as is normally the case, layer N-1 can utilize functions like segmentation, concatenation, and padding to make layer N-1 PDUs a fixed size.\",\n \"In the example discussed herein, a layer N PDU consisting of the actual signaling data 22 and the Integrity Check Info is discussed.\",\n \"The Integrity Check Info consists of the 32 bit MAC-I 26 and the message sequence number SN 24, which is needed at the peer end for the recalculation of MAC-I.\",\n \"The total length of the message 200 is then a combination of the signaling bits and the Integrity Check Info bits.\",\n \"At the receiving end the message is received including the signaling data part and the Integrity Check Info (which comprises the message sequence number SN and the 32-bit MAC-I).\",\n \"The signaling data together with the Integrity Check Info (ie the secret integrity the COUNT-I value, the direction of transmission, and the random number (FRESH)), are processed in a computing block, for example a function like the UTRAN f9 function, with a fixed function.\",\n \"A thus generated received computed message authentication code XMAC-I is then compared with the message authentication code MAC-I received in the transmitted message.\",\n \"If the two codes match (ie XMAC-I matches MAC-I), the recipient can trust that the message is intact, and the recipient then accepts the message.\",\n \"Otherwise, the message is discarded.\",\n \"The frame dependent COUNT-I number is actually the sum of a locally generated and incremented frame number HFN (Hyper Frame Number), which is added to the message sequence number, for example RRC_SN, and included in the message.\",\n \"The HFN is incremented each time SN reaches its maximum value (SN is normally very short, e.g.\",\n \"4 bits).\",\n \"As mentioned hereinabove the transmitted.block (layer N-1 PDU) normally has a fixed length.\",\n \"However, it may be that the signaling data bits together with the Integrity Check Info require more space than that provided in one layer N-1 PDU payload.\",\n \"One known way to deal with this problem is to segment the signaling message.\",\n \"In segmentation a signaling message, which is too long to fit in a single layer N-1 block, is passed on to a lower layer, where it is split up into two blocks (two layer N-1 PDUs), each with an appropriate layer N-1 header.\",\n \"Two blocks is just an example here, naturally a larger message may require even more than two blocks.\",\n \"If the second layer N-1 PDU is not totally filled with the layer N data, padding bits are inserted.\",\n \"At the receiving end before transferring to a higher layer, the two layer N-1 payloads are reassembled into one layer N PDU.\",\n \"To a person skilled in the art, it is immediately obvious that the use of padding bits is a potential waste of resources.\",\n \"TDMA systems, for example, have a limited radio block size, whereby a message including the full message authentication code does not necessarily fit into one radio block.\",\n \"This leads to the difficulty that the message has either to be sent without the MAC-I or in one or more additional segments.\",\n \"In addition, there are certain time critical messages, for example, handover messages, which must be sent in one radio block only.\",\n \"Generally, segmentation is not desirable, because it wastes radio resources and slows down the signaling procedure unnecessarily.\",\n \"One way to solve the above problem is to make the length of the message authentication code shorter than 32 bits.\",\n \"It has been proposed that such a message should include a field that defines the length of the message authentication code, a two-bit identifier, for example.\",\n \"This identifier allows certain discrete values: 8, 16, 24 and 32.This solution still has some problems.\",\n \"First, the identifier always takes two extra bits from the length of the message.\",\n \"Second, the discrete values are not flexible, and in some cases this can lead to the same problem as above, i.e.\",\n \"segmentation is needed for certain messages.\",\n \"A particularly advantageous technique for addressing the above stated problems is disclosed in Finnish patent application number FI20002453.This discloses a technique that allows the transmission of a message in a single lower layer data block even when the length of the message including the integrity check info exceeds the length of the lower layer data block.\",\n \"In all current known solutions, each message is authenticated separately.\",\n \"Each message contains a sequence number of the protocol it belongs to.\",\n \"Hence a binding between different steps in the existing solutions is achieved through the sequence number.\",\n \"The drawback to existing solutions is that at each step a 32-bit MAC is transmitted which takes a significant part of the signaling bandwidth.\",\n \"It is therefore an object to the present invention to provide an improved technique for taking the integrity of messages.\",\n \"SUMMARY OF THE INVENTION In accordance with the present invention there is provided a method of providing message authentication in a communication system comprising the steps of: transmitting a first message from a first device to a second device; transmitting a second message from the second device to the first device, the second message including a message authentication code determined using said first and second messages; transmitting a third message from the first device to the second device, the third messages including a message authentication code determined using the third message.\",\n \"In order to minimize the risk of ‘replay attacks’, the sequence numbers should preferably be maintained in the protocol.\",\n \"This is particularly important if the messages are short or otherwise small in number.\",\n \"The method may further comprise the step of storing the first message in the first device.\",\n \"The method may further comprise the steps of: responsive to receipt of the second message, determining an expected message authentication code using said first and second messages; and comparing the expected message authentication code to the received message authentication code.\",\n \"The method may further comprise the step of discarding the second message if the expected message authentication code does not match the received message authentication code.\",\n \"The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message; and comparing the expected message authentication code to the received message authentication code.\",\n \"The method may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.\",\n \"The third message may include a message authentication code determined using the third message and the second message.\",\n \"The method may further comprise the step of storing the second message in the second device.\",\n \"The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message and said second message; and comparing the expected message authentication code to the received message authentication code.\",\n \"The method according to claim 9 may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.\",\n \"The third message may include a message authentication code determined using the third message, the second message, and the first message.\",\n \"The method may further comprise the step of storing said first message in the second device.\",\n \"The method may further comprise the steps of: responsive to receipt of the third message, determining an expected message authentication code using said third message, said second message, and said first message; and comparing the expected message authentication code to the received message authentication code.\",\n \"The method may further comprise the step of discarding the third message if the expected message authentication code does not match the received message authentication code.\",\n \"The invention may thus advantageously reduce the bandwidth used for authentication.\",\n \"In a multi-step protocol, the invention provides for the messages exchanged at different steps to be grouped together for message authentication code computations.\",\n \"A message is saved by the sending party for subsequent verification at a later step.\",\n \"The number of required message authentication code computations and transmissions may be reduced to two independently of the number of steps in the authentication procedure.\",\n \"In three step signaling procedures, there is thus no need for a message authentication code to be computed in the first step.\",\n \"The invention is further advantageously applicable to procedures containing more than three messages.\",\n \"All communications that occur between two parties in various steps may be authenticated by the communicating parties in the last two steps of the protocol.\",\n \"The first and second devices are preferably elements of a mobile communication system.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS The invention will be described more closely with reference to the accompanying drawings, in which FIG.\",\n \"1 depicts the computation of a message authentication code; FIG.\",\n \"2 shows the contents of a message; FIG.\",\n \"3 illustrates a first embodiment of the present invention; FIG.\",\n \"4 illustrates a second embodiment of the present invention; FIG.\",\n \"5 depicts the creation of a message according to a preferred implementation; FIG.\",\n \"6 is a flow chart showing the creation of a message in the GERAN system, and FIG.\",\n \"7 is a flow chart of actions at the receiving end.\",\n \"DESCRIPTION OF PREFERRED EMBODIMENTS The invention is described herein by way of reference to particular non-limiting examples, and with particular reference to a GERAN system.\",\n \"The GERAN is specified by the Third Generation Partnership Project (3GPP).\",\n \"GERAN is an evolution of the GSM-system (Global System for Mobile Communication), the TDMA/136-system (Time Division Multiple Access System), and the EDGE-system.\",\n \"GERAN has no integrity protection of its own.\",\n \"Implementation of the same integrity algorithms used in UTRAN is suggested for a radio system using the GPRS/EDGE-radio connection network GERAN.\",\n \"This leads to certain significant problems, especially the problem of message segmentation.\",\n \"The present invention can be advantageously used for implementing an integrity algorithm in the GERAN system, and the present invention is described herein with reference to example implementations in such a system.\",\n \"However, the invention is not limited in its application to such a system.\",\n \"Radio interface protocols are needed to set up, reconfigure, and release Radio Bearer services.\",\n \"The protocol layers above the physical layer are called the data link layer (layer 2) and the network layer (layer 3).\",\n \"The control plane layer 2 contains two sub-layers: Medium Access Control (MAC) protocol and Radio Link Control (RLC) protocol.\",\n \"Layer 3 consists of one protocol, called Radio Recourse Control (RRC), which belongs to the control plane.\",\n \"The channels offered by the physical layer to the MAC layer are called logical channels.\",\n \"It shall be appreciated that the term ‘logical channel’ can be used for other purposes in other systems.\",\n \"For example, in the UTRAN the term logical channel referes to a channel offered by the MAC layer to higher layers.\",\n \"All higher layer signaling (mobility management, call control, session management, etc.)\",\n \"is encapsulated into RRC messages for transmission over the radio interface.\",\n \"The following provides a description of integrity protection for a message to be sent over a radio link.\",\n \"Referring to FIG.\",\n \"3, a first embodiment of the present invention is described.\",\n \"For the purpose of this illustrative example, it is assumed that messages are being exchanged between the mobile station 40 and a network 42.It is assumed that the mobile station 40 initiates the exchanged messages.\",\n \"The mobile station 40 constructs a message for transmission in the normal way in accordance with standard procedures.\",\n \"Prior to transmitting the message, as indicated by step 50, the mobile station 40 stores the initial message, labeled message_1, in its memory.\",\n \"As indicated by arrow 44, message_1 is then transmitted from the mobile station 40 to network 42.In accordance with the present invention, the first message is sent without a message authentication code.\",\n \"The network 42 receives the first message from the mobile station, and prepares a second message, message_2, for transmission back to the mobile station 40.In addition, in a step 52 the network 42 calculates a first message authentication code, MAC_I_1, using both the first message as received by the mobile station and the second message which it is to transmit to the mobile station.\",\n \"Thus the message authentication code to be transmitted with the second message is based on the combination of both the first and second messages.\",\n \"As indicated by arrow 46, the network 42 then transmits a second message, message_2, with the message authentication code MAC_I_1.Using the received second message, message_2, and the stored first message, message_1, the mobile station 40 in a step 54 calculates the expected message authentication code XMAC_I_1.If XMAC_I_1 is identical to MAC_I_1, then the mobile station 40 continues with message transmission.\",\n \"Otherwise the received message message_2 is discarded.\",\n \"In this embodiment of the invention, the mobile station 40 prepares to transmit a third message message_3 to the network 42.In the step 56 the mobile station 40 prepares a second message authentication code MAC_I_2 using the content of the third message message_3.As represented by arrow 58, the third message message_3 is transmitted to the network 42 together with the second message authentication code MAC_I_2.The network 42 then calculates the expected message authentication code XMAC_I_2 using the third message message_3 in step 58, and in the normal way compares this to the transmitted authentication code MAC_I_2.Whilst in FIG.\",\n \"3 the three messages are identified as message_1, message_2 and message_3, in actual signaling procedures they may be named, for example, XXX_request, XXX_command, XXX_complete (or “confirm”).\",\n \"Thus in accordance with the first embodiment of the invention, the message authentication code is left out of the first message.\",\n \"The message authentication code for the second message is calculated over the first and second messages, although the first message is not returned to the sender.\",\n \"The third message is integrity protected “normally”, with a message authentication code calculated over the third message itself.\",\n \"This technique ensures that the procedure cannot be used illegally by an intruder, even if the first message is not integrity protected.\",\n \"Referring to FIG.\",\n \"4, the second embodiment of the present invention is now described.\",\n \"Where appropriate, the same reference numerals as used in FIG.\",\n \"3 are used to refer to identical steps or procedures.\",\n \"The embodiment of the invention described with reference to FIG.\",\n \"4 is suitable for signaling procedures normally having either two or three messages.\",\n \"As in the embodiment described hereinabove with reference to FIG.\",\n \"3, the mobile station 40 prepares a first message message_1 for transmission to the network 42, and prior to transmitting it, as represented by arrow 44, stores it in a memory as represented by step 50.In step 52, the network 42 calculates a first message authentication code MAC_I_1 using both the first message and the second message.\",\n \"In an additional step 53, the network 42 saves the second message message_2 into its memory before transmission.\",\n \"As in the embodiment of FIG.\",\n \"3, the network 42 transmits the second message message_2 together with the first message authentication code MAC_I_1 as indicated by arrow 46 to the mobile station 40.In step 54, as in the embodiment of FIG.\",\n \"3, the mobile station 40 calculates the expected message authentication code XMAC_I_1 using both the received second message and the stored first message.\",\n \"If the expected message authentication code XMAC_I_1 is identical to the received message authentication code MAC_I_1, then the procedure is continued with.\",\n \"In this embodiment of the present invention, the mobile station 40 then calculates in step 55 a second message authentication code MAC_I_2 using both the third message to be sent, message_3, and the received second message, message_2.The mobile station 40 then transmits, as indicated by arrow 48, the third message, message_3, together with the message authentication code MAC_I_2.The network 42 then calculates the expected second message authentication code XMAC_I_2 using both the stored second message message_2 and the received third message message_3 in step 59.Thus, in common with the first embodiment described hereinabove with reference to FIG.\",\n \"3, the second embodiment as described with reference to FIG.\",\n \"4 similarly does not include any message authentication code in the first transmitted message.\",\n \"The difference of the second embodiment compared to the first embodiment is that the third message contains a message authentication code calculated over both the second and third messages, whereas in the first embodiment the message authentication code is calculated over only the third message.\",\n \"The technique of FIG.\",\n \"4 may be more advantageous in situations where the third message is added only for this purpose, ie it does not contain any actual information but is merely an acknowledgement message and is thus very short.\",\n \"From a security viewpoint, the calculation of the integrity check sum over a longer message is beneficial.\",\n \"The addition of a third message may be more preferable than segmentation of the first message, because the segmentation solution necessitates additional acknowledgement (on data link layer level).\",\n \"In a third embodiment of the present invention, the message authentication code transmitted with the third message is calculated over all three messages, ie message_1, message_2 and message_3.The advantage of this embodiment is that each check sum protects the maximum amount of the data communicated in the procedure.\",\n \"Thus, this variation excludes the possibility of an attack where a “man in the middle” replaces the first message with another one and modifies the message authentication code in the second message accordingly.\",\n \"Such an attack could only be successful if the attacker is able to modify the check sum correctly.\",\n \"There are two possible ways to do this: 1) the replay of an earlier message authentication code; or 2) pure guessing.\",\n \"The first way is only possible if the counter number repeats, which means the same integrity key has been in use for too long.\",\n \"The possibility of the second way being successful is a very low probability.\",\n \"However, if the message authentication code of the second message is not a full 32-bits long, the probability of the guessing attack becomes higher.\",\n \"If the “man in the middle” is to have any chance of making a successful attack when the third embodiment of the invention is in use, then it is necessary for either 1) or 2) to be succeeded twice, which is much more difficult.\",\n \"The present invention can particularly advantageously be used in combination with the technique described in Finnish patent application number FI120002453.Such a technique is described hereinbelow.\",\n \"It should be noted that the techniques described hereinbelow may only be utilized in steps where a message authentication code is being generated, and hence would not be used in accordance with the invention in relation to the generation and transmission of the first message.\",\n \"It should also be noted that other techniques may also be used.\",\n \"FIG.\",\n \"5 illustrates a situation where a signaling message 500 is to be sent in a secure manner over a lower layer radio link in one fixed length radio block, which can be a TDMA block, for example, without segmentation.\",\n \"The maximum block size allowed by the lower layer data block 501 is indicated by dotted lines in the figure.\",\n \"The signaling message without the Integrity Check Info (ICI) is in the illustrated situation shorter than the said maximum block size.\",\n \"This leads to a situation where the data to be sent either has to be segmented or sent without the message authentication code.\",\n \"Neither of these alternatives is acceptable.\",\n \"In order for the data to be sent in a sufficiently secure manner over a radio link, the computed message authentication code should be appended to it.\",\n \"However, it must be shortened in a predefined way (described in detail below).\",\n \"This truncated message authentication code diminishes the reliability of the integrity protection to some degree but still provides sufficient protection for the message.\",\n \"It should be noted, that the sequential number SN needed to form the COUNT-I parameter cannot be truncated.\",\n \"The message authentication code may be computed in the usual way in the device concerned and the MAC-I added with the message sequence number to the encoded message to form the actual PDU.\",\n \"Then the length of the message (without the Integrity Check Info) and/or the length of the PDU can be examined as follows.\",\n \"i) If the length of the message is longer than the length of the lower layer data block, the PDU is segmented into two or more data blocks as in prior art.\",\n \"ii) If the length of the PDU is shorter than the length of the lower layer data block, the PDU is placed into said lower layer data block and the rest of the block is filled with padding bits (normally by the lower layer itself).\",\n \"iii) If the length of the PDU is longer than the length of the lower layer data block but the extra bits are less than the size of the MAC-I, then the computed message authentication code is truncated so that the truncated PDU fits into one layer N-1 data block.\",\n \"However, truncation of the message authentication code diminishes the security of the message exchange.\",\n \"Therefore, the number of bits the MAC-I is allowed to be truncated by is limited to a certain maximum value, i.e.\",\n \"the truncated message authentication code has a certain minimum value.\",\n \"Thereafter, the truncated PDU is sent via a radio interface to the receiving end.\",\n \"At the receiving end the integrity is examined of the PDU received.\",\n \"First, the part including the signaling bits and the part including the Integrity Check Info are separated.\",\n \"Then a message authentication code is recomputed based on exactly the same algorithm and using the same parameters as were used at the transmitting end.\",\n \"The message authentication code of the received message is then compared with the recomputed authentication code.\",\n \"FIG.\",\n \"6 shows as a flowchart a more detailed example of one implementation of the method according to the invention from the point of view of the transmitting end.\",\n \"At stage 600 a time critical RRC message is to be sent through a radio interface, for example from the network to a mobile.\",\n \"Most signaling messages sent between a mobile station MS and the network, for example, must be integrity protected.\",\n \"Examples of such messages are RRC, MM, CC, GMM, and SM messages.\",\n \"Integrity protection is applied at the RRC layer, both in the mobile station and in the network.\",\n \"Integrity protection is usually performed for all RRC (Radio Recourse Control) messages, with some exceptions.\",\n \"These exceptions can be: 1.messages that are assigned to more than one recipient, —messages that have been sent before integrity keys were created for the connection, 2.frequently repeated messages, including information which does not need integrity protection.\",\n \"The message is encoded according to the specified message transfer syntax at stage 601.The encoded message (bit string) is called here E. A 32-bit message authentication code MAC-I, which is to be added to the encoded message, is calculated at stage 602.The message authentication code not only depend on the encoded message but also on several other parameters.\",\n \"The following input parameters are needed for calculation of the integrity algorithm: the encoded message, the 4-bit sequence number SN, the 28-bit hyper-frame number HFN, the 32-bit random number FRESH, the 1-bit direction identifier DIR, and the most important parameter—the128-bit integrity key IK.\",\n \"The short sequence number SN and the long sequence number HFN together compose the serial integrity sequence number COUNT-I.\",\n \"When the message authentication code is computed using the UMTS integrity algorithm and the above parameters, it is guaranteed that no one other than the actual sender can add the correct MAC-I code to the signaling message.\",\n \"COUNT-I, for example, prevents the same message from being sent repeatedly.\",\n \"However, if the same signaling message for some reason or other is to be sent repeatedly, the MAC-I code differs from the MAC-I code that was in the previously sent signaling message.\",\n \"The aim of that is to protect the message as strongly as possible against eavesdroppers and other fraudulent users.\",\n \"Due to the fact that a TDMA radio block has a fixed length, the length of the message has to be checked to avoid segmentation of the message.\",\n \"The RRC layer makes a decision as to whether the segmentation of the message concerned is to be allowed or not.\",\n \"At stage 603 the total length of the signaling message to be sent without the message authentication code is calculated using the following formula: X=max_size−sizeof(E)−sizeof(RRC_SN) In the above formula max_size is the maximum size (in bits) of a RRC message that can be sent in one radio block (i.e.\",\n \"there is no need for segmentation).\",\n \"Sizeof(E) is the size (in bits) of the encoded message and sizeof(RRC_SN) is the size of the RRC sequence number, a 4-bit working assumption.\",\n \"X defines the length (in bits) of the rest of the fixed length message, which is still left after the minimum number of bits are reserved for the message authentication code, the untruncated MAC-I size may be different.\",\n \"Next, at stage 604, a comparison is made to ascertain whether the calculated X is between values 0 and min_MACI_len, where the latter value is the minimum allowed length for the message authentication code.\",\n \"This minimum length is a predefined value, which can be either the same for all messages or even a message type specific value.\",\n \"It is clear that the smaller the value, the weaker the protection.\",\n \"So it is obvious that a minimum length must be determined so that the message can be sent with adequate security.\",\n \"If the answer after said comparison is YES, this means that the message authentication code does not fit with the signaling message to be sent in one radio block.\",\n \"In other words, the space left in one radio block is too short even for a shortened message authentication code after the signaling message is put into the block.\",\n \"If this is the case, the system protocol defines 605 as the next action to be carried out.\",\n \"If the answer after comparison is NO, the next step is to compare whether X is between values min_MACI_len and 32, stage 606.If X is between those values, i.e.\",\n \"if the answer after the comparison in stage 604 is YES, then X bits of the message authentication code with the RRC_SN are added to the encoded message, stage 607.The sequence number RRC_SN is needed for integrity protection, that is, for calculation of the message authentication code at the receiving end.\",\n \"Note that the MAC-I size is also 32 bits in the UTRAN system.\",\n \"In some other systems, the ‘normal’ MAC-I size may be something different.\",\n \"The length of the message authentication code is shortened in a predefined way.\",\n \"Thus, the size of the message authentication code transmitted over the radio path depends dynamically on the size of each encoded message, not on the type of the message.\",\n \"The decisions are made at the RRC layer as to the minimum message authentication code size and as to when the size of the said code may be shortened.\",\n \"In some cases the RRC layer can make the decision that the message authentication code is not to be shortened even though this would have been possible.\",\n \"Such cases might occur when strong protection is demanded for a message.\",\n \"The next step 611 is to send the message including the integrity protection info (E+MAC-I+RRC_SN) to the lower layers for transmission over the radio interface to the mobile station.\",\n \"If the answer in the above comparison 604 is NO, a final comparison is made as to whether the value of X is greater than 31, stage 608.If the answer is YES, this means that neither shortening nor segmentation is needed.\",\n \"Now the whole message authentication code and the RRC_SN are appended to the encoded message E, stage 609.The next step is stage 611.If the answer to the comparison at stage 608 is NO, i.e.\",\n \"if X is smaller than 0, which means that the size of the encoded message sizeof(E) is greater than the maximum size of the RRC message max_size, then two different alternatives A and B, are possible at stage 610: 1.add the entire MAC-I (+RRC_SN) to the message; 2.set sizeof(E)=(sizeof(E)−max_size) and rerun the previous steps.\",\n \"Which of the two above alternatives is selected depends on the protocol according to the system.\",\n \"Alternative A means that the whole message authentication code with the sequence number RRC_SN is added to the block, since the message has to be segmented anyway.\",\n \"Thus with this alternative, it is not important whether adding Integrity Protection Info causes additional segmentation or not.\",\n \"In alternative B, the attempt is made to avoid the additional segmentation caused by the addition of Integrity Check Info.\",\n \"Thus the sizeof(E) is set one full data block shorter than what was given in stage 601, and the truncation algorithm for the MAC-I is rerun starting from step 603.FIG.\",\n \"7 shows as a flowchart an example of one implementation of the method according to the invention from the point of view of the receiving end.\",\n \"At stage 700 the receiving end gets a Service Data Unit (SDU) comprising the signaling data M from the lower layers.\",\n \"It is assumed here that this message is the same as in the previous example in FIG.\",\n \"6.The next step is that the part including signaling data bits and the part including the Integrity Check Info MAC-I (the message authentication code with the sequence number the RRC_SN) are separated and a message authentication code with RRC sequence number RRC_SN is decoded in stage 701.The actual message (signaling data) can still be stored as an encoded bit string at this point.\",\n \"In prior art systems the message received is discarded immediately if the received and the recomputed message authentication codes do not match.\",\n \"But according to the described embodiment of the invention, the receiving end first examines the length of the message authentication code and, depending on the result, it then decides, if and how the message is to be processed further.\",\n \"For example, if the receiving end finds that the message authentication code received is shorter than it should be, it may assume that the code has been truncated.\",\n \"Instead of n bits the truncated code comprises m bits.\",\n \"If the truncation exceeds the predetermined maximum amount known by the receiving end, the message is discarded.\",\n \"If truncation does not exceed the predetermined maximum amount as known by the receiving end, the bits of the truncated message authentication code are compared bit-by-bit to the bits of the recomputed authentication code of full length.\",\n \"When the m bits of the truncated message authentication code match the corresponding bits of the recomputed message authentication code, the integrity check of the message received is passed.\",\n \"The length of the message authentication code of the message received is examined at stages 702 and 706.In addition, the length of the entire message (including the signaling data and the integrity check info) received is also examined at stage 704.At stage 702 a check is made as to whether the length of the MAC-I is ‘normal’, in this example 32 bits.\",\n \"If YES (the answer to stage 702 is NO), the flow proceeds to stage 703, where the message is processed in the normal way.\",\n \"The message authentication code is checked, the message is decoded, etc.\",\n \"using the same algorithm and parameters as were used at the transmitting end.\",\n \"Provided that stage 702 yields the YES alternative, meaning that the MAC-I has been truncated, a check is made as to whether the length of the message is a multiple of the max_size (sizeof(M) mod max_size—0), justifying the MAC-I truncation.\",\n \"A NO alternative yields a protocol error, for which reason the message received is discarded 705.A YES answer after stage 704 leads to stage 706, when a comparison is made as to whether the length of the message authentication code is greater than or equal to the minimum allowed MAC-I length (min_MACI_len).\",\n \"A NO alternative yields a protocol error, for which reason the received message is discarded 707.If the length is greater than or equal to the minimum value, the transmitting end may have shortened the MAC-I code in the correct way.\",\n \"With the integrity key and all the other needed parameters, the expected message authentication code XMAC-I is calculated 708, using the same algorithm as for the transmitting end.\",\n \"The calculated XMAC-I has to be truncated in order to compare its size with the size of the transmitted MAC-I, stage 709.If the truncated XMAC-I does not correspond to the transmitted truncated MAC-I 710, an integrity error is found and the received message is discarded 711.If the result of the comparison is positive, the message is decoded 712.The final check 713 is made after decoding the actual signaling data 712.The final check is to find out whether there are some padding bits in the received message.\",\n \"Since the MAC-I has been truncated due to message size, no padding bits are allowed (since the padding bits should have been used for the MAC-I).\",\n \"The Integrity check is OK whenever no RRC padding bits are found 713, i.e.\",\n \"it is then ensured that the message has been sent from the authorized party 715.Otherwise, a protocol error is found and the message is discarded, stage 714.The above defined preferred technique for generation of the message authentication code may be used in the embodiments of the present invention at those steps where a message authentication code is generated.\",\n \"An implementation and embodiment of the present invention has been explained above with some examples.\",\n \"However, it is understood that the invention is not restricted to the details of the above embodiment and that numerous changes and modifications can be made by those skilled in the art without departing from the characteristic features of the invention.\",\n \"The embodiment described is to be considered illustrative but not restrictive.\",\n \"Therefore, the invention should be limited only by the attached claims.\",\n \"Thus, alternative implementations defined by the claims, as well as equivalent implementations, are included in the scope of the invention.\",\n \"For example, instead of at the RRC layer the decision concerning the MAC-I size can be made at some other layer, e.g.\",\n \"the RLC layer.\",\n \"In that case, the RLC must know whether segmentation of the message(s) in the transmission buffer is to be allowed or not.\",\n \"The protocol layers from top to bottom may be, for example, RRC, LLC (Logical Link Control), LAPDm (Link Access Protocol on the Dm channel), PDCP (Packet Data Convergence Protocol), RLC, MAC (Medium Access Control Protocol), and PHY (Physical Layer) In addition, the minimum value, which is set at min_MACI_len might also depend on the signaling message used.\",\n \"The grouping of signaling messages into different min_MACI_len categories can be carried out either simply according to the message type.\",\n \"Grouping can be based on other factors as well, such as on whether the message is so that for critical signaling messages the value is greater than for non-critical signaling messages.\",\n \"For some non-critical messages the min_MACI_len could be set as low as 8 bits, for example.\",\n \"Utilised in combination with the techniques described with reference to FIGS.\",\n \"5 to 7 the present invention may allow, in certain situations, a longer MAC-I, thereby further improving integrity.\",\n \"The present invention is independent of the length of the message authentication code.\",\n \"Such code may be longer or shorter than 32 bits.\",\n \"The length may also differ between messages in one procedure.\",\n \"It should also be noted that although this application is made only from the signaling standpoint, integrity protection can also be applied in some systems to the user plane data.\",\n \"The same principles and methods described in this application are applicable also for user plane data packets, although the actual protocol layers performing the integrity protection, and the message authentication code truncation would then be different.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10476988"}}},{"rowIdx":992,"cells":{"text":{"kind":"list like","value":[["Shared water reservoir beverage system","An apparatus for making and dispensing a plurality of beverages, the dispensing apparatus comprising; a plurality of beverage dispensing modules (32), each beverage module having a container (68) and a brewing assembly (30) for making a respective beverage and delivering the respective beverage to the container, and a housing (34) associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid for supply to the beverage making assemblies to make respective beverages."],["1.An apparatus for brewing and dispensing a plurality of beverages, the dispensing apparatus comprising: (a) a plurality of beverage dispensing modules, each beverage dispensing module having a container and a brewing assembly for brewing a respective beverage and delivering the respective beverage to the container; and (b) a housing associated with and positioned externally of the plurality of beverage dispensing modules and defining a chamber containing a heated liquid for supply to the brewing assemblies to brew the respective beverages.","2.The apparatus of claim 1 wherein each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.","3.The apparatus of claim 1 further comprising a plurality of conduits, each conduit communicating with the chamber for supplying heated liquid from the chamber to the brewing assembly of a respective beverage dispensing module.","4.The apparatus of claim 1 wherein each of the beverage dispensing modules has a front and a back, the backs facing the housing.","5.The apparatus of claim 4 wherein the fronts of the beverage dispensing modules face the same direction generally outwardly away from the housing.","6.The apparatus of claim 1 wherein the housing is positioned behind the beverage dispensing modules.","7.The apparatus of claim 1 wherein the beverage dispensing modules have fronts and further comprising a plurality of second beverage dispensing modules having fronts, the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules facing opposite directions.","8.The apparatus of claim 7 wherein the housing is positioned between the beverage dispensers and the second beverage dispensers.","9.The apparatus of claim 1 further including a heating element positioned at least one of proximate to and in the chamber for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.","10.The apparatus of claim 1 further including a controller for selectively activating the brewing of the respective beverages and a plurality of conduits for supplying liquid from the chamber to the brewing assemblies of the plurality of modules.","11.The apparatus of claim 10 further including a plurality of activation switches associated with the beverage dispensing modules, each activation switch adapted to send a switch signal to the controller to activate brewing of the respective beverage at the corresponding beverage dispensing module.","12.The apparatus of claim 11 further including a liquid sensor for sensing the level of the liquid in the housing and for sending a liquid level signal to the controller to activate brewing of the respective beverage.","13.The apparatus of claim 11 further including a temperature sensor for sensing the temperature of the liquid in the housing and for sending a temperature signal to the controller to activate brewing of the respective beverage.","14.The apparatus of claim 11 further including a heating element associated with the housing for receiving a heating signal from the controller to activate heating of the liquid within the chamber.","15.The apparatus of claim 11 further including an inlet valve for providing liquid to the housing in response to a valve signal from the controller.","16.The apparatus of claim 10 further including a plurality of activation switches associated with the plurality of beverage modules, a liquid level sensor for sensing the level of the liquid in the housing, a temperature sensor for sensing the temperature of the liquid in the housing, and a heating element for heating the liquid in the housing, the controller adapted to activate brewing of the respective beverage in response to activation of a respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.","17.The apparatus of claim 1 further including a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.","18.The apparatus of claim 1 further including a liquid flavor dispensing assembly for selectively dispensing a plurality of different flavorings to the brewing assemblies of at least some of the beverage dispensing modules so that at least some of the beverages have different flavors.","19.An apparatus for brewing and dispensing a plurality of beverages, the dispensing apparatus comprising: (a) a plurality of beverage dispensing modules, each beverage dispensing module having a container, a brewing assembly for brewing a respective beverage and delivering the respective beverage to the container, and a dispensing valve for dispensing the respective beverage from the beverage dispensing module; (b) a housing associated with the plurality of beverage dispensing machines and defining a chamber containing a heated liquid for supply to the brewing assemblies to brew the respective beverages; and (c) a plurality of conduits communicating with the chamber and with a corresponding making assembly for supplying heated liquid from the chamber to the brewing assemblies of the plurality of beverage dispensing modules.","20.The apparatus of claim 19 wherein the beverage dispensing modules face the same direction generally outwardly away from the housing and the housing is positioned behind the beverage dispensing modules.","21.The apparatus of claim 19 further including a controller for selectively activating the brewing of the respective beverages.","22.The apparatus of claim 21 further including a plurality of activation switches associated with the plurality of beverage modules, a liquid level sensor for sensing the level of the liquid in the housing, a temperature sensor for sensing the temperature of the liquid in the housing, and a heating element for heating the liquid in the housing, the controller adapted to activate brewing of the respective beverage by a respective beverage module in response to activation of a respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.","23.An apparatus for brewing and dispensing a plurality of beverages, the dispensing apparatus comprising: (a) a plurality of beverage dispensing modules, each beverage dispensing module having a container and a brewing assembly for brewing a respective beverage and delivering the respective beverage to the container; (b) a housing associated with the plurality of beverage dispensing machines and defining a chamber containing a heated liquid for supply to the brewing assemblies to brew the respective beverages; (c) a plurality of conduits for supplying liquid from the chamber to the brewing assemblies of the plurality of beverage dispensing modules; and (d) a controller for selectively activating the brewing of the respective beverages.","24.The apparatus of claim 23 wherein each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage."],["<SOH> BACKGROUND <EOH>This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.","Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.","In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.","Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.","As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.","Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.","Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.","Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.","These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.","To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.","In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process."],["<SOH> SUMMARY <EOH>The present disclosure relates to apparatus for making and dispensing beverages.","The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.","Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.","The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.","In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.","In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.","The apparatus may also include a plurality of conduits for supplying liquid to the assembly.","The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.","Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.","The housing may be positioned behind the beverage dispensing modules.","In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.","With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.","The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.","A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.","The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.","The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.","In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.","In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.","Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived."],["BACKGROUND This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.","Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.","In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.","Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.","As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.","Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.","Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.","Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.","These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.","To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.","In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process.","SUMMARY The present disclosure relates to apparatus for making and dispensing beverages.","The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.","Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.","The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.","In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.","In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.","The apparatus may also include a plurality of conduits for supplying liquid to the assembly.","The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.","Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.","The housing may be positioned behind the beverage dispensing modules.","In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.","With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.","The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.","A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.","The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.","The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.","In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.","In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.","Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived.","BRIEF DESCRIPTION OF THE DRAWINGS The detailed description particularly refers to the accompanying figures in which: FIG.","1 is a partial fragmentary side elevational view of a modular beverage dispensing apparatus; FIG.","2 is a top plan view of the modular beverage dispensing apparatus of FIG.","1; FIG.","3 is a side elevational view of the modular beverage dispensing apparatus of FIGS.","1 and 2; FIG.","4 is a schematic illustration of the modular beverage dispensing apparatus; FIG.","5 is a schematic illustration of one module of the modular beverage dispensing apparatus; FIG.","6 is a diagrammatic illustration of a dispensing and disposal valve configuration; FIG.","7 is a partial fragmentary top plan view taken along 7-7 in FIG.","6; FIG.","8 is a diagrammatic illustration of a liquid flavor distributing system; FIGS.","9 and 10 are diagrammatic illustrations of the arrangement of liquid flavor distributing systems; FIG.","11 is a cross sectional view, schematic in nature, of a portion of a powdered beverage, soluble beverage dispensing system including connections to dispense liquid flavor to the soluble beverage production stream; FIGS.","12 and 13 are diagrammatic illustrations of the liquid flavor distributing system coupled to the soluble beverage dispensing system of FIG.","11; FIG.","14 is a perspective view of a soluble beverage dispenser having a slide-out assembly; FIG.","15 is an enlarged perspective view of the soluble beverage dispenser of FIG.","14 showing a user removing a container from the soluble beverage dispensing device using the slide-out assembly; FIGS.","16 and 17 are enlarged views of portions of the slide-out assembly of the soluble beverage dispenser of FIG.","14; FIG.","18 is a perspective view of a cable and spring leverage system which assists movement of the slide-out assembly relative to the beverage dispensing system; FIGS.","19 and 20 are schematic illustrations of a communication link between a base station and a beverage dispensing device; and FIGS.","21-28 are schematic illustrations of the circuitry associated with the communication system of FIGS.","19 and 20 AND INCLUDE FIG.","22A, FIGS.","22-22D, FIGS.","23-23D, FIG.","24, FIGS.","24A-24B, FIGS.","25A-25B, FIGS.","26A-26, FIG.","27, FIGS.","27A-27C, FIG.","28 and FIGS.","28A-28E.","DESCRIPTION OF THE DRAWINGS While the present disclosure may be susceptible to embodiment in different forms, there is shown in the drawings, and herein will be described in detail, embodiments with the understanding that the present description is to be considered an exemplification of the principles of the disclosure and is not intended to limit the disclosure to the details of construction and the arrangements of components set forth in the following description or illustrated in the drawings.","A beverage dispensing apparatus 30 illustrated generally in FIGS.","1-4 as comprising a plurality of beverage dispensing modules 32 and a housing 34 that defines a chamber 36 containing water or other liquid suitable for use in brewing.","The housing 34 desirably acts as a central reservoir for a constant supply of heated water to be used by the beverage dispensing modules 32 to brew several different beverages.","The beverage dispensing apparatus 30 may have any suitable construction and configuration.","The beverage apparatus 30 may include any suitable number of beverage dispensing modules 32 oriented in any suitable manner, depending on various circumstances, such as, for example, customer demand, customer usage, available store space, and desired aesthetics.","Similarly, the housing 34 may have any suitable construction and configuration that desirably is consistent with customer demand, customer usage, available store space and desired aesthetics.","A plurality of conduits in the form of water lines 38 provide flow communication between the housing 34 and the beverage dispensing modules 32 to supply water for brewing.","In a preferred embodiment, the housing 34 is positioned behind the beverage dispensing modules 32 and centrally located relative to the beverage dispensing modules 32 to minimize the distance between the housing and each of the beverage dispensing modules, thereby increasing operational efficiency and minimizing space.","Terms including beverage and beverage making as used herein are intended to be broadly defined as including but not limited to the brewing of coffee, tea and any other brewed beverage, including a final beverage or food product as well as producing an intermediate product to be combined with a final beverage or food product.","This broad interpretation is also intended to include, but is not limited to any process of infusing, steeping, reconstituting, diluting, dissolving, saturating or passing a liquid through or otherwise mixing or combining a beverage substance with a liquid such as water without a limitation to the temperature of such liquid unless specified.","This broad interpretation is also intended to include, but is not limited to beverage substances such as ground coffee, tea, liquid beverage concentrate, powdered beverage concentrate, freeze dried coffee or other beverage concentrates, to obtain a desired beverage or other food.","In the illustrated embodiment, for example, the beverage dispensing apparatus 30 comprises ten beverage dispensing modules 32 oriented in a generally U-shaped manner that substantially surrounds the housing 34.As illustrated, four of the beverage dispensing modules 32 face the same direction, four other beverage dispensing modules 32 face an opposite direction, and two of the beverage dispensing modules 32 are positioned in a space-efficient manner consistent with a desired orientation.","A simplified form of the beverage apparatus 30 is shown in FIG.","4 which uses simplified block diagram illustrations to show multiple beverage dispensing modules 32 which are configured to provide a beverage brewing system.","The beverage apparatus 30 shown in FIG.","4 is very similar to a soluble beverage apparatus 30a also shown in FIG.","4.The beverage apparatus 30, as will be described in greater detail herein below, relates to a beverage brewing or producing system whereas the soluble beverage apparatus 30a, as will be described in greater detail herein below, relates to a soluble or powdered beverage dispensing system.","Both beverage apparatuses 30, 30a employ a common heated water reservoir contained in housing 34 to which the individual modules 32, 32a are connected.","As shown in FIGS.","2-4, the housing 34 desirably is centrally located to serve multiple beverage modules 32, 32a.","The benefit of a common housing 34 functioning as a common reservoir to supply water, instead of individual tanks for each module 32, 32a, is that it substantially reduces the cost and greatly increases the efficiency of the system.","Also it reduces the redundancy of multiple heated water reservoirs, reduces the wattage required by the system and increases flexibility in the number of modules 32, 32a which might be included.","The particular configuration with the housing 34 positioned in close proximity to or between several modules 32, 32a is beneficial in that it also minimizes the length of water lines 38 connecting the housing 34 to the modules 32, 32a.","Additionally, a larger body of water in the housing 34 tends to help maintain the temperature and reduce temperature loss over time as well as maintaining a larger volume of brew water at a desired temperature.","While the housing 34 retained within the beverage apparatus 30 is insulated to reduce heat loss and reduce heat load on the ambient environment, heat which may escape from the housing 34 will generally pass to the modules 32, thereby increasing the efficiency of maintaining the temperature of the beverages retained in the modules 32.Insulation desirably is provided to reduce heat loss to the ambient environment.","This will also help reduce the exertion on the ambient air-conditioning system thereby further increasing efficiency of the present beverage apparatus 30, 30a.","With reference to FIGS.","1-4, each module 32, 32a produces a separate brewed beverage.","The centralization of a housing 34 helps reduce the individual cost associated with the module 32,32a and improves the ease of manufacturing of the modules.","Also, there is increased commonality of parts associated with this type of system.","The modular configuration of the beverage apparatus 30 using the modules 32, 32a increases the potential variability of the physical size and brewer configuration of the system.","This is beneficial to accommodate the needs of a particular location.","In other words, only a desired or required number of modules 32, 32a will be needed at any given time.","If a location has an increase in demand for brewed or soluble beverages additional modules 32, 32a may be added as necessary for the particular change in demand.","Additionally, with regard to repair and maintenance, the beverage apparatus 30, 30a can be designed such that individual modules 32, 32a can be removed for repair and maintenance as necessary with an identical module being inserted in the missing modules.","In a preferred embodiment, (FIG.","5) the housing 34 includes a heating element 52, and one or more system sensors such as a level sensor 86 and a temperature sensor 84.Such components tend to be generally readily available and easily replaced and as such further enhance the reliability of the overall beverage apparatus 30, 30a.","With reference to FIG.","3, the line 38 connecting each housing 34 to a brewing portion 40 of the module 32 is shown.","Water 42 retained in the housing 34 can be readily dispensed.","A water inlet line 44 is provided to introduce unheated water 50 to the housing 34.The inlet line 44 runs from an upper portion 46 of the housing 34 and directs the inward water downwardly towards a bottom portion 48 of the housing 34.Introducing the unheated water 50 towards the bottom of the reservoir introduces the unheated water 50 in close relation to a heated element 52 thereby accelerating the heating of the water.","Additionally, the introduction of unheated water 50 in the bottom portion 48 tends to force heated water in the upper portion 46 upwardly.","As such, the water 42 in the upper portion 46 tends to be the hottest water in the housing 34.Introducing unheated water 50 from the upper portion has the benefit of providing an entrance into the reservoir through a reservoir wall 56 with little or no pressure on the entry therethrough.","This may result in an increase in the reliability of the entry therethrough.","Additionally, a reinforcing or strengthening structure 58 is provided extending between opposed walls 60, 62 of the housing 34.This helps prevent bowing or stress on the walls 60, 62 which might otherwise result from the force of the water on the walls.","Multiple reinforcing structures 58 may be provided throughout the housing 34 as necessary to provide support for the size and dimension of the housing required for a particular system.","FIG.","5, for example, shows a diagrammatic illustration of an embodiment of the apparatus 30 shown in FIGS.","1-4.The beverage dispensing modules 32 can have any suitable construction desirably adapted to produce beverages, store beverages for ready consumption, and facilitate ready dispensing of the beverages.","Each module 32 includes a brewing assembly 30 that may have any suitable construction that can be used to produce beverage such as brewing coffee or other beverages.","The illustrated brewing assembly 30, for example, includes a brewing substance retainer or assembly 64 including a funnel 66.Such a retainer 64 may be well known in the art and may include a filter (not shown) retained in the funnel 66 for holding a charge of beverage substance.","The funnel 66 may be positioned above a reservoir or container portion 68 of the dispensing module 32 which receives brewed beverage from the funnel.","A dispensing assembly 70 in the form of a controllable valve may be associated with the container 68 for dispensing beverage from the module 32.With reference to FIG.","5 a simplified review of the beverage brewing process is described.","A brew actuator switch 72 desirably is associated with each of the modules 32.To initiate a brew cycle for one of the modules 32 a respective brew activation switch 72 is operated.","Operation of the switch 72 sends a signal over line 74 to controller 76.Controller 76 operates an inlet valve 78 over line 80 to introduce water to the housing 34.Also, heating element 52 is coupled to the controller 76 over line 82.The temperature sensor 84 and level sensor 86 are coupled to the controller over lines 88, 90 respectively.","If the controller 76 detects proper level and temperature it will initiate a brew cycle thereby displacing water from the housing 34 through a sprayhead assembly 92.The sprayhead assembly 92 communicates with the substance retainer 64 and dispenses water into the funnel 66 for producing a beverage therein.","The illustrated module 32 includes a level sensor 94 associated with the container 68 which is coupled to the control over line 96.The level sensor can detect a change in level in beverage retained in the container 68 which may be used to activate a brew cycle.","Also, the level detector 94 may sense the level in the beverage container 68 thereby indicating if the container is full or above the desired level to prevent initiation of a brew cycle.","This will prevent over flow of the beverage server 68.The beverage dispensing modules 32 can be constructed to dispense beverage in any suitable manner.","In a preferred embodiment, for example, a dispenser or faucet switch 98 may be associated with the dispensing assembly 70 and coupled with a dispensing handle or control 100.Activation of the control 100 will activate the switch 98 over line 102 connected to the controller 76 thereby indicating that the controller 76 should operate a solenoid valve 104 (FIGS.","6 and 7), via line 106 to dispense beverage therefrom.","As will be described in greater detail below with regard to FIGS.","6 and 7, a disposal or dump valve 108 is also coupled with the container 68 to facilitate controllable disposal, via line 110 coupled to the controller 76.With reference to FIGS.","6 and 7, the beverage container 68 may further include a manifold 114.The illustrated manifold 114 includes a primary passage 116 which communicates with an interior cavity or reservoir 118 of the container 68.Beverage flows from the cavity 118 through the primary passage 116.The solenoid valves 104, 108 communicate with the passage 116.A dispensing passage 120 and a disposal passage 122 communicate with the primary passage 116.Solenoid valve 104 and 108 desirably are normally closed and require specific activation in order to open the valves.","During a dispense cycle the switch or control 100, for example a controllable faucet as shown in FIG.","5, is activated thereby activating solenoid valve 104.When the dispense solenoid valve 104 is opened beverage 124 will flow from the cavity through the solenoid valve 104.Under this condition disposal valve 108 is closed.","When beverage must be disposed of, the disposal solenoid valve 108 is opened while maintaining the dispense solenoid valve 104 in a closed position.","This will allow the disposal of beverage through a separate dispensing line.","Disposal of beverage may occur as a result of a time lapsed period whereby after a specified period of time beverage should be disposed of.","For example, if a beverage only maintains a desired flavor profile or other characteristics for a period of time, for example 2 hours, at the end of two hours after the beverage brewing cycle is initiated, the beverage may be disposed of through the disposal valve 108.Such disposal may also be used at the end of a service cycle such that at the end of the day all beverages may be disposed of at a predetermined time.","The disposal valve may also be used in a manner to flush and cleanse the beverage brewing system.","In this regard, at a predetermined period of time, the controller 76 may operate the disposal valve 108 to dispose of any beverage retained in the container 68.The disposal valve 108 and dispense valve 104 may be closed.","The controller 76 may then dispense a quantity of water into the cavity 118 thereby flushing and sanitizing the reservoir.","After holding the heated water for a predetermined period of time the water may be disposed through the disposal valve 108.Multiple rinsing or flushing cycles may be controlled by the controller 76.In this regard, both the cavity 118, the container 68 and the associated passages 116, 122, 120 and solenoid valve 104, 108 maybe cleansed and sanitize periodically and automatically.","It will be appreciated that these automatic cycles will likely greatly increase the cleanliness, taste reliability and sanitation of such beverage dispensing systems as such procedures require considerable amount of time on the part of manual operators.","The use of controllable solenoid valves 104, 108 also prevents dispensing of beverage during a brew cycle.","For example, once a brew cycle is initiated the controller 76 can lock out the solenoid valves 104, 108.This will allow for the brewing cycle to complete and brewed beverage to mix within the container 68 to insure consistent flavor.","The beverage dispensing modules 32 can be constructed in any other suitable manner to dispense brewed beverage in any suitable manner in accordance with various embodiments disclosed.","If desired, for example, the beverage containers 68 of the beverage dispensing modules 32 can instead be in the form of carafes (not shown) or have any other suitable construction.","In a preferred embodiment, during a brewing cycle, heated water 134 is dispensed from the housing 34 through the water dispensing system.","Heated water 134 is used to infuse a beverage brewing substance 136 retained in the brewing assembly 64.An opening 138 is provided in the bottom of the funnel 66 to allow a brewed beverage 140 to flow therefrom.","As shown, for example, in previous figures, a brewed beverage container 68 is provided to collect the beverage 140.The beverage container 68 may be of any suitable form.","With reference to FIG.","8, a liquid flavor dispensing system 130 may be employed with the modular beverage dispensing system disclosed.","The liquid flavor dispensing system 130 can have any suitable configuration and can be used to supply flavoring 132 to all or less than all of the beverage dispensing modules 32.The liquid dispensing system 130 may also be used to supply a different flavoring 132 to each of the different beverage dispensing modules 32.Additionally, the flavorings 132 can be supplied to the beverage dispensing modules by a single liquid flavor dispensing system 130 or several liquid flavor dispensing systems.","Further, the flavoring 132 may in the form of the typical flavorings associated with brewed coffee or instead be in any other suitable form such as, for example, honey, peach, lemon, herbal flavors, or any other suitable flavoring which might be desired by a beverage consumer for any number of different beverages.","In a preferred embodiment, a small quantity of flavoring 132 is introduced into the water which is dispensed to the beverage brewing substance 136.The beverage brewing substance may be in the form of decaff or regular ground coffee as well as tea substances or other substances which might be used in an infusion beverage brewing process.","The objective in using the liquid flavoring material 132 is to introduce a small quantity of concentrated material or substance 132 during the brewing process so that the flavor becomes fully mixed with and saturated in the heated water 134.Introduction of the flavor 132 early in the brewing process will help assure that the flavor fully mixes with the water, is fully flushed through the beverage brewing substance to maximize the value of the flavor and is used to produce an aroma which might entice purchasers of the beverage as part of the beverage purchasing experience.","The flavor could also be dispensed into the beverage brewing substance and subsequently mixed with the water and the resultant brew during the brewing process.","A benefit of adding flavors at the time of brewing and/or mixing soluble beverages, over using pre-flavor beverage substances, is the increased effectiveness of the flavorings.","The increased effectiveness of the flavorings reduces the amount of flavoring required to produce a desired taste profile.","Under these circumstances, the flavorings are mixed with the beverage substance under optimal conditions to maximize the flavoring benefits of the flavorings.","For example, the time prior to mixing the flavoring with the brewing substance is nominal thereby virtually eliminating any degradation of the flavoring which might occur if the flavoring was exposed to air.","A liquid flavor dispensing system 130 is illustrated diagrammatically in FIG.","8.With this embodiment, the liquid flavor dispensing system dispenses a different liquid flavoring 132 in a controlled manner to several of the brewing assemblies of the beverage dispensing modules 32.The flavor can be dispensed to a water or liquid distribution system which controllably dispenses water to a beverage making substance, the beverage malting substance or both.","With reference to FIGS.","8-10, the illustrated liquid flavor dispensing system 130 desirably includes a plurality of packages or containers 144a-144h, each of which retains a respective liquid flavor 132 therein.","The containers 144 desirably are in controlled communication with respective beverage dispensing modules 32a-32d in any suitable manner.","For example, a section of tubing or other passage 146 may connect each container 144 to a respective check valve 148.The check valve 148 is part of a respective solenoid or other form of pump device 150 coupled to the controller 76 via a respective line 152.A respective small diameter tube 154 extends from the solenoid pump 150 to a respective second check valve or flavor injection port 156 at a respective terminal end 158 of the tube 154.A respective sensor 160 can be coupled to the controller via a respective line 162 to monitor and detect a low level of the respective flavoring 132 in the respective container 144.As will be described in greater detail below with regard to sensing and monitoring, the status of the flavorings 132 can be reported by the controller 176.The check valves 148 and 156 at each end of each of the tubes 154 provide a sealed line to which the container 144 can be attached.","Also, either alternatively or in addition to the use of the sensor 160, the flavor usage can be inferred based on information collected on a characteristic of the pump such as operation or usage for example, pump actuation.","For example, a sensor may be associated with the pump and run time of the pump 150 could be monitored and a calculation made to infer the amount of flavorings remaining in a corresponding flavorings container 144.The system would provide a signal to the operator, such as “checlc-status” when the level of flavorings approaches, attains or drops below a predetermined level, for example 20% remaining.","Such advance warning would give the operator the opportunity to change the flavorings container 144 before it runs out.","The advance warning also helps assure that a reasonable margin of error in the inferred calculation is considered and accommodated for.","In use, when a brewing cycle is initiated, the solenoid pump 150 can be activated simultaneous with the initiation of the brewing cycle of one of the beverage dispensing modules or shortly thereafter.","A quantity, for example 9 ml for use in brewing ½ gallon of coffee, of liquid flavoring 132 is pumped from the container 144 through the tubing 154.The check valve or flavor injection port 156 is positioned in close proximity to a sprayhead 170 of the dispensing assembly 92.Dispensing of the flavor 132 into the water will allow the flavor to mix with the heated water for brewing substance during the saturation and extraction process.","Generally, it is desirable to use the flavoring at the start of the cycle which will result in completely integrating the flavor during the brewing cycle.","The process may be repeated with other beverage dispensing modules 32, 32a and, if desired, with other flavors.","The flavor injection ports 156 may, for example, include a check valve of a type as produced by Smart Products.","This is a positive pressure inverted check valve which requires positive pressure through the tube 154 to open the valve.","Positioning of the injection port 156 in close proximity to the sprayhead 170 results in the benefit of clearing and rinsing the surface as a result of the water vapor and heat during a brewing cycle.","Periodic manual cleaning of the sprayhead 170 and injection port 156 helps maintain a clean system.","Desirably, because the injection port 156 is an inverted pressure check valve, physical contact with the tip of the injection port 156 through which the liquid 132 is dispensed does not result in dispensing of liquid 132.Rather, physical contact with the end of the injection port 156 causes the valve to close as opposed to open.","The system also prevents the entry of atmosphere into the container 144 or line 154 thereby preventing contamination or degrading of the flavor or quality of the flavorings.","This system provides a generally closed system which prevents the flavorings contact with the atmosphere and prevents contamination providing a generally aseptic environment.","With reference to FIG.","11, the liquid flavor dispensing system 130 as described with regard to FIGS.","8-10 may be used with a soluble beverage system 176.In FIG.","11, the liquid dispensing system dispenses liquid flavor into a hot water path 174 of a soluble or powdered beverage dispensing system 176.The devices used to dispense the liquid flavor are the same as those describe above with regard to FIG.","8.As such, only the terminal ends 158a, 158b and 158c of multiple lines 154a, 154b, and 154c are shown.","Similarly, the multiple injection ports 156a, 156b, and 156c are shown.","The soluble beverage dispensing system 176 retains a quantity of powdered beverage mix (not shown) in a beverage dispensing container 178.The dispensing of soluble beverage material in powder form is generally known in the art.","The container 178 includes a dispensing auger (not shown) which controllably dispenses powder by operation of a dispensing drive or motor 180 controlled by the controller 76a.","Powdered beverage material is dispensed through the passage 182 into a mixing chamber 184.The hot water line 174 communicates with the mixing chamber 184 to combine with the powder.","The combined powder and hot water flow into an agitating chamber 186 there below.","A whipper motor 188 and agitating blade 190 further blend the powder and water combination for dispensing a beverage 192 through a nozzle 194.The flavor dispensing system 130 (see, FIG.","8) controllably dispenses liquid flavor 132 from respective containers 144 using corresponding pumps via the lines 154a, 154b, and 154c.","Either individual flavors or combinations of flavors may be dispensed into the hot water line.","Such dispensing may be programmed in the controller 76a.","Positioning of the injection ports 156a, 156b, 156c in the heated water path helps to assure that the liquid flavors are rinsed from the injection ports during and at the conclusion of the heated water dispensing cycle during a beverage dispensing cycle.","It should be noted that generally these powdered beverage dispensing systems have a momentary hot water overrun at the end of the cycle during which powdered beverages dispense from the container 178.The slight overrun of heated water helps to assure that the mixing chamber 184, whipping chamber 186, and nozzle 194 are rinsed clean at the end of the cycle.","The injection ports 156a, 156b and 156c have been positioned in the hot water line to provide the rinsing characteristics and benefits of the overrun of hot water.","Furthermore, it is commonly known to have at least one daily rinse cycle in which heated water is dispensed through the assembly 176 to maintain cleanliness and sanitation of the system.","During a brew cycle as shown in FIG.","8, liquid flavor is generally preferably dispensed at the initiation of the brew cycle.","As noted, this helps to ensure complete mixing and optimal extraction of the flavor from the beverage brewing substance 136.A predetermined volume of liquid flavor 132 can be dispensed into the brewing assembly 30 at the initiation of the brew cycle.","In contrast, the soluble system 176 as shown in FIG.","11 may have a variable quantity of beverage 192 dispensed therefrom.","It is anticipated that multiple beverage sizes may be dispensed from the soluble system 176.As a result, the liquid flavoring 132 must be dispensed at a rate consistent with the concentration flavor profile or recipe, and powder also dispensed by the system.","For example, if a consumer wishes to obtain a 12 ounce beverage the proper amount of liquid flavoring must be dispensed for a 12 ounce beverage.","However, the consumer may be allowed to control whether they wish to have a full 12 ounces of beverage or if they decide to short the beverage resulting in, perhaps, only 10 ounces being dispensed.","As such, the pump associated with the powdered beverage system 176 must be calibrated to dispense small quantities of liquid flavoring 132 on a per unit time basis to assure a desirable and consistent mixing of flavor with the powder.","This will help assure that the desired flavor is achieved regardless of the quantity of beverage dispensed.","The solenoid pump 150 used with the liquid flavor dispensing system 130 is a precision controlled metering pump.","It is anticipated that other forms of the metering pump may be used with this system.","However, in the present embodiment, the metering pump is controllably operated to dispense very small quantities of the liquid flavor 132.The frequency of the plunger rate, the stroke of the plunger as well as the duration of plunger operation the solenoid pump 150 can be controlled by the controller 76, 76a.","As a result, very small quantities of highly concentrated liquid flavor can be dispensed into the beverages 140, 192 during the corresponding beverage preparation process.","As such, precise quantities can be added to provide flexibility and tailoring of the resultant beverages.","The system 130 can be calibrated using volumetric guidelines associated with various beverage recipes and flavor materials to match particular flavor preferences, for example for various demographics.","As such, a franchise operation can custom blend and configure the flavor of beverages dispensed by the beverage dispensing system to meet and satisfy regional preferences.","Additionally, if preferences change over time the system can be modified and calibrated to match those preferences.","This adds further flexibility to the beverage system disclosed.","With reference to FIGS.","14-18, a powdered or soluble beverage dispensing system 199 is shown.","While a soluable beverage system 199 has been shown and described as a powder dispensing system, it is envisioned that such a soluable system also includes a liquid concentrate system which dispenses a quality of liquid coffee concentrate for mixing with water or other dilution or reconsituting substance.","Reference to a soluable beverage system is intended to be broadly interpreted.","Such a beverage dispensing system has been generally and diagrammatically shown in FIG.","11.With further reference to FIG.","14, the container 178 is shown positioned inside a housing 200.The passage 182 coupled to the container 178 is shown positioned for dispensing into the mixing chamber 184 and communicating with the whipping chamber 186 and nozzle 194.A portion of the housing 200 in the form of a door 202 conceals the powdered beverage system 176 in the housing 200.Also, the liquid flavor dispensing system 130 as described hereinabove may be used with the powdered beverage or soluble beverage dispensing system 176 illustrated in FIGS.","14-18.One of the potential difficulties involved with a soluble beverage dispensing system is that the container 178 must be removed periodically for refilling with powdered or soluble beverage substance as well as regular maintenance and cleaning.","The container 178 may not be very heavy when it is empty because it is generally formed of a plastic material.","However, once filled, it may contain 5 to 10 pounds of powder material.","With the container 178 being positioned in an elevated location and containing a quantity of material, it may be difficult to lift the container into a desired position in the housing 200.The container 178 is positioned in an upper position in the housing 200 in order to benefit from gravity assistance when dispensing powdered beverage therefrom to the mixing chamber 84.Additionally, it is important to align the dispensing passage 182 with the housing 184 to help assure that when powdered beverage material is dispensed from the container 178 it flows into the proper path for mixing with water.","As such, it would be desirable to provide an apparatus and system for facilitating improved ease of removal and replacement of the powdered beverage container 178 relative to the housing 200.As shown in FIGS.","14-18, a container positioning assembly or container positioner 220 in the form of a slide-out support or shelf is provided.","The container positioning assembly 220 includes a pair of handles 222 which can be grasped (see FIG.","15) by a user to extract the container 178 attached to the assembly 220 for removing it from the housing 200.As shown in FIG.","15, the assembly 220 allows the container 178 to be pulled straight out of the housing 200 thereby allowing disengagement of the passage 182 relative to the mixing chamber 184.Movement of the assembly 220 also replaces the container 178 in a desired position to align the dispensing passage 182 with the mixing chamber 184.As can be seen in FIGS.","15-17, the container 178 has been removed from the housing 200 in order to refill the container 178 but does not have to be removed from the shelf 250 of the assembly 220.Support rails 226 are positioned on each side of container 178 and are engaged with side walls 228 of the housing 200 (FIG.","15).","Each of the guide rails 226 includes a pair of tracking slots 230, 232 which are engaged with corresponding upper 234 and lower 236 guide rollers.","A support roller 240 is positioned along an exterior edge 242 of the guide 226 to further help facilitate smooth and controlled movement of the assembly 220.Support rollers 240 and guide rollers 234, 236 are mounted on the sidewalls of the housing 200.The guides 226 are connected by a cross extending shelf 250 on which the container 178 is positioned.","The combination of the guide rails 226 and rollers help the assembly 220 slide outwardly and inwardly relative to the housing 200.FIG.","16 shows only the portion of the guide rails 226 extending past the edge of container 178.The guide rollers 240, 234, 236 associated with the guides 226 prevent tipping of the shelf 250 and associated container 178.When the assembly 220 is stowed in the housing 200 the shelf 250 rests on a corresponding surface 252 of the housing 200.As can be seen in FIGS.","15-17, the guides 226 include a generally horizontal portion 254 and a vertically oriented portion 256.The shelf 250 and container 178 are associated with the generally horizontal portion 254.The generally vertical portion 256 helps to allow forward movement as well as downward movement of the assembly 220 relative to the housing 200.The guide including the portion 254, 256 help facilitate movement of the container into and out of the housing 200 and forward and downward movement.","In the embodiment as shown, the assembly facilitates positioning of the container 178 approximately 6-8 inches downwardly from its stored position in the housing 200.This lowering of the container 178 is beneficial in that it allows for easy filling of the container.","Also, by fixing the container to the shelf 250 and the assembly 200 it prevents spilling or otherwise tipping of the container 178 during filling or other processes.","This can result in savings in terms of lost beverage material as well as cleanup costs and associated complications.","In the stored position, detents 260, 262 are provided on the guides 230, 232, respectively.","The detents 260, 262 allow the rollers 234, 236 respectively, to be engaged therewith to prevent casual disengagement of the assembly 220 from the housing 200.This helps to lock or retain the assembly 220 in a desired stored or stowed position in the housing 200.As the assembly 220 is moved outwardly from the housing 200, the track guides 234, 236 track along the corresponding channels 230, 232 generally horizontally along the horizontal portion 254.As such, the container 278 attached to the shelf 250 sides forwardly and outwardly from the housing.","At a position where the generally horizontal portion 254 transitions into the partially vertical portion 256 (elbow 270) the guides allow the assembly to slide downwardly and forwardly.","In the extended most position the guides 234, 236 engage corresponding extended detents 272, 274, respectively.","These detent positions 272, 274 help retain the assembly 220 in the outward or extended most position.","The assembly 220 also includes an assist system 280 which includes cables 282 and biasing members to springs 284 forming a cable-biasing assembly help facilitate removal of and replacement of the container 178 relative to the housing 200.With reference to FIG.","18, the assist system 280 includes a spring 284 attached at one 286 to a back portion of the housing and at a second end to a first pulley 288.The cable 282 is attached at a first end 290 to a corresponding portion of the housing and extends through the first pulley 288 and up and around a second pulley 292.The second end 294 of the cable 282 attaches to a corresponding portion of the guides 226 at attachment point 296.Attachment of the cable 282 to the guide 226 is shown in FIG.","16.The assist assembly 288 helps further facilitate ease of removal and replacement of the container 178 relative to the housing 200.For example, when the container 178 is removed from the housing on the assembly 220 the spring 284 stores energy and is stretched as the cables 282 are extended.","The cables 282 are extended as a result of the forward and downward movement of the guides 226.Extension of the springs 284 expends the stored energy and imparts a return force in the springs which helps reduce the force required to return the container 178 to the housing 200.This is particularly useful when an empty container is removed from the housing, filled with powdered material and then returned.","Returning the filled container 178 normally would require additional effort on the part of the operator and as such the return forces in the assist system 280 help make the return easier.","Turning now to FIGS.","19 and 20, the beverage system also includes a communications system or means 300 for communicating 302 between a corresponding beverage brewing, beverage making apparatus or beverage dispensing modules 32, 32a and a receiver or base station 304.Each of the modules 32, 32a includes a controller 76, 76a which is centralized for use with multiple modules or dedicated to an individual module.","The controller collects information about the module or modules.","For purposes of this present discussion, we will refer to the controller as being dedicated to multiple modules and receiving information from each individual module associated therewith.","The many modules associated with a single control also provides the added benefit of the modular assembly as described hereinabove.","It should also be noted that the details of the circuitry used to achieve the communication system are illustrated in the detailed schematics provided in FIGS.","21-28 and all corresponding sub-portions thereof.","For example, FIG.","21 is a general schematic of the overall system.","FIGS.","22-28 provide details about the general system shown in FIG.","21.FIG.","21 has been noted with cross reference to the other figures which provide details about the portions of the circuitry shown generally in FIG.","21.The controllers 76, 76a include the link 302 which connects the controller to the corresponding receiver or base station 304.In the present embodiment as diagrammatically illustrated in FIGS.","19 and 20, the controllers 76, 76a communicate with the receiver 304 by way of an RF link 302.Generally, the controllers are remote from the receiver and use the Link 302 and the associated means for linking to communicate the collected information about the beverage making apparatus.","Antennae 306, 308 are provided on the controllers 76, 76a and the corresponding receivers 304 respectively.","It should be noted that the communication link 302 may be in any one of a variety of forms such as hardwired physical point-to-point link, optical lines, light wave, ultrasonic, infrared or any other form of communication link between one or more devices.","The receiver 304 receives information from the controller or various components of the modules 32, 32a to identify information relating to the modules and operation of the modules.","The receiver 304 may be in the form of a monitoring unit positioned in an appropriate location relative to an operator or attendant who has responsibility for or otherwise maintains the modules 32, 32a.","The receiver 304 includes a means for displaying display 305 and can provide visual auditor or other information about numerous conditions.","The display 304 allows a form of the collected information about the beverage making apparatus to be displayed at the receiver.","The visual display may include, but is not limited to lights, text, symbolic images (i.e., dispenser “full” showing a colored or shaded dispenser and a dispenser “empty” showing an unshaded dispenser), and mechanical devices that tare operated or other visual displays.","An auditory display may include, but is not limited to speech information, alarms, tones or other signals that van be heard.","Tactile displays are also contemplated in the form of a vibrating surface, vibrating device worn by the operator or other means for letting the operator or other means for letting the operator know about he collected information.","For example, the following is a list of the messages or conditions which can be displayed: Receiver fault; Please wait attempting communication; Communication failure, coffee module #; Communication fault, soluble module #; Flavor [type] low, coffee module #; Flavor [type] low, soluble module #; Hopper low, soluble module #; Hopper empty, soluble module #; Coffee empty, coffee module #; Coffee low, coffee module #; Freshness expired, coffee module #; Dumping in ______ minutes, coffee unit #; Server removed, coffee module #; Refill too long; Temperature probe open; Temperature probe short; Heating too long; Over flow safety; Coffee stations all OK; and Soluble stations all OK For example, in an convenience store setting, multiple modules 32, 32a may be generally remotely positioned in one portion of the store, for example, a rearward location of the store, if desired.","The operator of the beverage system may also be the same person responsible for stocking of the equipment, and register activities.","As such, one person is required to operate and maintain many components of the convenience store facility.","The receiver 304 may be positioned at the cash register so that while the attendant is operating the cash register he may also be notified of matters that require his attention at the modules 32, 32a.","For example, if a module 32a indicates that a container 68 is empty the controller 76 will communicate with the receiver 304 to indicate this condition.","The receiver may also be configured with audible alarms and visual displays to provide additional information.","For example, a display may identify which module and the specific condition associated with the module which has provided a signal to the receiver 304.The operator can then make a decision how to use that information relative to his responsibilities at the cash register.","If an emergency requires immediate attention he can secure the cash register and address the emergency while attending to the module 32.Alternatively, if it is a slow period in the store and a module is indicating that a container 68 is empty he can put off attending to this condition until he completes one or more transactions in queue.","The signal communication between the controller 76 and the receiver 304 may be a single path signal or in the form of a multiple path verification signal.","For example, as shown in FIGS.","19 and 20 a first signal 310 is transmitted from the controller 76 to the receiver 304.This initial communication may report the status of a condition at the module 32.The receiver 304 can then return a copy of the information 312 to the controller 76.If the controller confirms the information received (312) it can then send a return confirmation 314 that the information is correct.","This is a useful communication link in the present situation due to the presence of electrical noise and other interference within other beverage system applications.","For example, each of the components associated with the modules 32, 32a may provide some degree of electrical or other noise.","Additionally, other systems within the beverage system setting may be producing RF or other signals which could create interference.","As such, the present embodiment providing the verification communication links 310-314 helps facilitate and assure accurate and timely communication.","In the event of excessive interference such as from patrons of the store using telephone or RF communication devices, the controller 76 can continue to periodically send a message 310 to the receiver 304 until the verification signal 312 is received.","While preferred embodiments are disclosed, illustrated and described, it is envisioned that those skilled in the art may devise various modifications and equivalents without departing from the spirit and scope of the disclosure as recited in the following claims."]],"string":"[\n [\n \"Shared water reservoir beverage system\",\n \"An apparatus for making and dispensing a plurality of beverages, the dispensing apparatus comprising; a plurality of beverage dispensing modules (32), each beverage module having a container (68) and a brewing assembly (30) for making a respective beverage and delivering the respective beverage to the container, and a housing (34) associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid for supply to the beverage making assemblies to make respective beverages.\"\n ],\n [\n \"1.An apparatus for brewing and dispensing a plurality of beverages, the dispensing apparatus comprising: (a) a plurality of beverage dispensing modules, each beverage dispensing module having a container and a brewing assembly for brewing a respective beverage and delivering the respective beverage to the container; and (b) a housing associated with and positioned externally of the plurality of beverage dispensing modules and defining a chamber containing a heated liquid for supply to the brewing assemblies to brew the respective beverages.\",\n \"2.The apparatus of claim 1 wherein each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.\",\n \"3.The apparatus of claim 1 further comprising a plurality of conduits, each conduit communicating with the chamber for supplying heated liquid from the chamber to the brewing assembly of a respective beverage dispensing module.\",\n \"4.The apparatus of claim 1 wherein each of the beverage dispensing modules has a front and a back, the backs facing the housing.\",\n \"5.The apparatus of claim 4 wherein the fronts of the beverage dispensing modules face the same direction generally outwardly away from the housing.\",\n \"6.The apparatus of claim 1 wherein the housing is positioned behind the beverage dispensing modules.\",\n \"7.The apparatus of claim 1 wherein the beverage dispensing modules have fronts and further comprising a plurality of second beverage dispensing modules having fronts, the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules facing opposite directions.\",\n \"8.The apparatus of claim 7 wherein the housing is positioned between the beverage dispensers and the second beverage dispensers.\",\n \"9.The apparatus of claim 1 further including a heating element positioned at least one of proximate to and in the chamber for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.\",\n \"10.The apparatus of claim 1 further including a controller for selectively activating the brewing of the respective beverages and a plurality of conduits for supplying liquid from the chamber to the brewing assemblies of the plurality of modules.\",\n \"11.The apparatus of claim 10 further including a plurality of activation switches associated with the beverage dispensing modules, each activation switch adapted to send a switch signal to the controller to activate brewing of the respective beverage at the corresponding beverage dispensing module.\",\n \"12.The apparatus of claim 11 further including a liquid sensor for sensing the level of the liquid in the housing and for sending a liquid level signal to the controller to activate brewing of the respective beverage.\",\n \"13.The apparatus of claim 11 further including a temperature sensor for sensing the temperature of the liquid in the housing and for sending a temperature signal to the controller to activate brewing of the respective beverage.\",\n \"14.The apparatus of claim 11 further including a heating element associated with the housing for receiving a heating signal from the controller to activate heating of the liquid within the chamber.\",\n \"15.The apparatus of claim 11 further including an inlet valve for providing liquid to the housing in response to a valve signal from the controller.\",\n \"16.The apparatus of claim 10 further including a plurality of activation switches associated with the plurality of beverage modules, a liquid level sensor for sensing the level of the liquid in the housing, a temperature sensor for sensing the temperature of the liquid in the housing, and a heating element for heating the liquid in the housing, the controller adapted to activate brewing of the respective beverage in response to activation of a respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.\",\n \"17.The apparatus of claim 1 further including a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.\",\n \"18.The apparatus of claim 1 further including a liquid flavor dispensing assembly for selectively dispensing a plurality of different flavorings to the brewing assemblies of at least some of the beverage dispensing modules so that at least some of the beverages have different flavors.\",\n \"19.An apparatus for brewing and dispensing a plurality of beverages, the dispensing apparatus comprising: (a) a plurality of beverage dispensing modules, each beverage dispensing module having a container, a brewing assembly for brewing a respective beverage and delivering the respective beverage to the container, and a dispensing valve for dispensing the respective beverage from the beverage dispensing module; (b) a housing associated with the plurality of beverage dispensing machines and defining a chamber containing a heated liquid for supply to the brewing assemblies to brew the respective beverages; and (c) a plurality of conduits communicating with the chamber and with a corresponding making assembly for supplying heated liquid from the chamber to the brewing assemblies of the plurality of beverage dispensing modules.\",\n \"20.The apparatus of claim 19 wherein the beverage dispensing modules face the same direction generally outwardly away from the housing and the housing is positioned behind the beverage dispensing modules.\",\n \"21.The apparatus of claim 19 further including a controller for selectively activating the brewing of the respective beverages.\",\n \"22.The apparatus of claim 21 further including a plurality of activation switches associated with the plurality of beverage modules, a liquid level sensor for sensing the level of the liquid in the housing, a temperature sensor for sensing the temperature of the liquid in the housing, and a heating element for heating the liquid in the housing, the controller adapted to activate brewing of the respective beverage by a respective beverage module in response to activation of a respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.\",\n \"23.An apparatus for brewing and dispensing a plurality of beverages, the dispensing apparatus comprising: (a) a plurality of beverage dispensing modules, each beverage dispensing module having a container and a brewing assembly for brewing a respective beverage and delivering the respective beverage to the container; (b) a housing associated with the plurality of beverage dispensing machines and defining a chamber containing a heated liquid for supply to the brewing assemblies to brew the respective beverages; (c) a plurality of conduits for supplying liquid from the chamber to the brewing assemblies of the plurality of beverage dispensing modules; and (d) a controller for selectively activating the brewing of the respective beverages.\",\n \"24.The apparatus of claim 23 wherein each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage.\"\n ],\n [\n \"<SOH> BACKGROUND <EOH>This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.\",\n \"Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.\",\n \"In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.\",\n \"Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.\",\n \"As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.\",\n \"Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.\",\n \"Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.\",\n \"Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.\",\n \"These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.\",\n \"To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.\",\n \"In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process.\"\n ],\n [\n \"<SOH> SUMMARY <EOH>The present disclosure relates to apparatus for making and dispensing beverages.\",\n \"The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.\",\n \"Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.\",\n \"The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.\",\n \"In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.\",\n \"In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.\",\n \"The apparatus may also include a plurality of conduits for supplying liquid to the assembly.\",\n \"The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.\",\n \"Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.\",\n \"The housing may be positioned behind the beverage dispensing modules.\",\n \"In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.\",\n \"With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.\",\n \"The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.\",\n \"A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.\",\n \"The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.\",\n \"The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.\",\n \"In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.\",\n \"In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.\",\n \"Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived.\"\n ],\n [\n \"BACKGROUND This disclosure relates to beverage systems and, more particularly, to a beverage apparatus for preparing, dispensing, monitoring, controlling and, if desired, flavoring beverages.\",\n \"Over the past 10 years, the coffee and beverage industry has experienced an evolution in connection with retail sales of fresh coffee.\",\n \"In particular, there is now a significant customer demand for individual servings of fresh coffee at, among other places, coffee shops, convenience stores, and fast food restaurants.\",\n \"Associated with the increase in demand for retail purchases of individual servings of fresh coffee are increases in demand for flavored coffee and for different types of flavored coffee as well as a variety of other beverages, such as teas, cocoas, etc.\",\n \"As a result of these significant changes in the coffee and beverage industry, the service market has changed drastically over the past 10 years.\",\n \"Coffee shops in which consumers can walk in and purchase a fresh cup of coffee have substantially increased in number.\",\n \"Convenience food stores, fast food restaurants and other retailers now devote substantial floor space to serving customer demand.\",\n \"Moreover, these changes place increasing demands on employees of retail outlets to monitor the coffee brewing and beverage preparation equipment to ensure that there is a constant supply of fresh coffee available to consumers and in many cases to also ensure that there is a sufficient number of different flavors of fresh coffee available.\",\n \"These changes in the coffee service industry have also created an increased need for efficiency in serving the consuming public.\",\n \"To be competitive in this expanding marketplace, efficiency in the brewing, storage, and dispensing of coffee is increasingly important.\",\n \"In particular, there is now a premium on being able to serve in an efficient manner the large demand for not only fresh coffee but also different flavors of coffee, in light of the substantial retail space needed for the coffee brewing and dispensing equipment and the continuous responsibilities in administering the brewing and dispensing process.\",\n \"SUMMARY The present disclosure relates to apparatus for making and dispensing beverages.\",\n \"The apparatus comprises a plurality of beverage dispensing modules, and a housing associated with the plurality of beverage dispensing modules and defining a chamber containing a liquid.\",\n \"Each beverage dispensing module has a container and an assembly for producing a respective beverage and delivering the respective beverage to the container.\",\n \"The liquid contained in the chamber is supplied to the assemblies to produce the respective beverages.\",\n \"In a preferred embodiment, each beverage dispensing module includes a dispensing valve for dispensing the respective brewed beverage from the beverage dispensing module.\",\n \"In a preferred embodiment, the apparatus may also include a controller for selectively activating the production of the respective beverages.\",\n \"The apparatus may also include a plurality of conduits for supplying liquid to the assembly.\",\n \"The dispensing modules and housing may have any suitable construction and desirably are oriented in a manner that is space efficient and facilitates easy administration and usage of the apparatus.\",\n \"Desirably, the beverage dispensing modules have a front and a back, the backs facing the housing, and the fronts may face the same direction.\",\n \"The housing may be positioned behind the beverage dispensing modules.\",\n \"In accordance with one embodiment, for example, the apparatus also includes a plurality of second beverage dispensing modules and the fronts of the beverage dispensing modules and the fronts of the second beverage dispensing modules face opposite directions.\",\n \"With this embodiment, desirably the housing is positioned behind the beverage dispensers and behind the second beverage dispensers.\",\n \"The apparatus may further include a heating element for heating the liquid within the housing, a level sensor for sensing the level of the liquid within the housing, and a temperature sensor for sensing the temperature of the liquid within the housing.\",\n \"A plurality of activation switches desirably are associated with the beverage dispensing modules, each activation switch is adapted to send a switch signal to the controller to activate production of the respective beverage.\",\n \"The controller preferably is adapted to activate production of the respective beverage in response to activation of the respective activation switch, a liquid level signal from the liquid level sensor, and a temperature signal from the temperature sensor.\",\n \"The apparatus also may include an inlet valve for providing liquid to the housing in response to a valve signal from the controller.\",\n \"In addition to being able to produce and serve a plurality of respective beverages, the apparatus desirably also includes a liquid flavor dispensing assembly for dispensing flavoring to the brewing assembly of at least one of the beverage dispensing modules to flavor the respective brewed beverage.\",\n \"In a preferred embodiment, for example, the liquid flavor dispensing assembly selectively dispenses a plurality of different flavorings to the assemblies of at least some of the beverage dispensing modules to provide beverages having different flavors.\",\n \"Additional features will become apparent to those skilled in the art upon consideration of the following detailed description of drawings exemplifying the best mode as presently perceived.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS The detailed description particularly refers to the accompanying figures in which: FIG.\",\n \"1 is a partial fragmentary side elevational view of a modular beverage dispensing apparatus; FIG.\",\n \"2 is a top plan view of the modular beverage dispensing apparatus of FIG.\",\n \"1; FIG.\",\n \"3 is a side elevational view of the modular beverage dispensing apparatus of FIGS.\",\n \"1 and 2; FIG.\",\n \"4 is a schematic illustration of the modular beverage dispensing apparatus; FIG.\",\n \"5 is a schematic illustration of one module of the modular beverage dispensing apparatus; FIG.\",\n \"6 is a diagrammatic illustration of a dispensing and disposal valve configuration; FIG.\",\n \"7 is a partial fragmentary top plan view taken along 7-7 in FIG.\",\n \"6; FIG.\",\n \"8 is a diagrammatic illustration of a liquid flavor distributing system; FIGS.\",\n \"9 and 10 are diagrammatic illustrations of the arrangement of liquid flavor distributing systems; FIG.\",\n \"11 is a cross sectional view, schematic in nature, of a portion of a powdered beverage, soluble beverage dispensing system including connections to dispense liquid flavor to the soluble beverage production stream; FIGS.\",\n \"12 and 13 are diagrammatic illustrations of the liquid flavor distributing system coupled to the soluble beverage dispensing system of FIG.\",\n \"11; FIG.\",\n \"14 is a perspective view of a soluble beverage dispenser having a slide-out assembly; FIG.\",\n \"15 is an enlarged perspective view of the soluble beverage dispenser of FIG.\",\n \"14 showing a user removing a container from the soluble beverage dispensing device using the slide-out assembly; FIGS.\",\n \"16 and 17 are enlarged views of portions of the slide-out assembly of the soluble beverage dispenser of FIG.\",\n \"14; FIG.\",\n \"18 is a perspective view of a cable and spring leverage system which assists movement of the slide-out assembly relative to the beverage dispensing system; FIGS.\",\n \"19 and 20 are schematic illustrations of a communication link between a base station and a beverage dispensing device; and FIGS.\",\n \"21-28 are schematic illustrations of the circuitry associated with the communication system of FIGS.\",\n \"19 and 20 AND INCLUDE FIG.\",\n \"22A, FIGS.\",\n \"22-22D, FIGS.\",\n \"23-23D, FIG.\",\n \"24, FIGS.\",\n \"24A-24B, FIGS.\",\n \"25A-25B, FIGS.\",\n \"26A-26, FIG.\",\n \"27, FIGS.\",\n \"27A-27C, FIG.\",\n \"28 and FIGS.\",\n \"28A-28E.\",\n \"DESCRIPTION OF THE DRAWINGS While the present disclosure may be susceptible to embodiment in different forms, there is shown in the drawings, and herein will be described in detail, embodiments with the understanding that the present description is to be considered an exemplification of the principles of the disclosure and is not intended to limit the disclosure to the details of construction and the arrangements of components set forth in the following description or illustrated in the drawings.\",\n \"A beverage dispensing apparatus 30 illustrated generally in FIGS.\",\n \"1-4 as comprising a plurality of beverage dispensing modules 32 and a housing 34 that defines a chamber 36 containing water or other liquid suitable for use in brewing.\",\n \"The housing 34 desirably acts as a central reservoir for a constant supply of heated water to be used by the beverage dispensing modules 32 to brew several different beverages.\",\n \"The beverage dispensing apparatus 30 may have any suitable construction and configuration.\",\n \"The beverage apparatus 30 may include any suitable number of beverage dispensing modules 32 oriented in any suitable manner, depending on various circumstances, such as, for example, customer demand, customer usage, available store space, and desired aesthetics.\",\n \"Similarly, the housing 34 may have any suitable construction and configuration that desirably is consistent with customer demand, customer usage, available store space and desired aesthetics.\",\n \"A plurality of conduits in the form of water lines 38 provide flow communication between the housing 34 and the beverage dispensing modules 32 to supply water for brewing.\",\n \"In a preferred embodiment, the housing 34 is positioned behind the beverage dispensing modules 32 and centrally located relative to the beverage dispensing modules 32 to minimize the distance between the housing and each of the beverage dispensing modules, thereby increasing operational efficiency and minimizing space.\",\n \"Terms including beverage and beverage making as used herein are intended to be broadly defined as including but not limited to the brewing of coffee, tea and any other brewed beverage, including a final beverage or food product as well as producing an intermediate product to be combined with a final beverage or food product.\",\n \"This broad interpretation is also intended to include, but is not limited to any process of infusing, steeping, reconstituting, diluting, dissolving, saturating or passing a liquid through or otherwise mixing or combining a beverage substance with a liquid such as water without a limitation to the temperature of such liquid unless specified.\",\n \"This broad interpretation is also intended to include, but is not limited to beverage substances such as ground coffee, tea, liquid beverage concentrate, powdered beverage concentrate, freeze dried coffee or other beverage concentrates, to obtain a desired beverage or other food.\",\n \"In the illustrated embodiment, for example, the beverage dispensing apparatus 30 comprises ten beverage dispensing modules 32 oriented in a generally U-shaped manner that substantially surrounds the housing 34.As illustrated, four of the beverage dispensing modules 32 face the same direction, four other beverage dispensing modules 32 face an opposite direction, and two of the beverage dispensing modules 32 are positioned in a space-efficient manner consistent with a desired orientation.\",\n \"A simplified form of the beverage apparatus 30 is shown in FIG.\",\n \"4 which uses simplified block diagram illustrations to show multiple beverage dispensing modules 32 which are configured to provide a beverage brewing system.\",\n \"The beverage apparatus 30 shown in FIG.\",\n \"4 is very similar to a soluble beverage apparatus 30a also shown in FIG.\",\n \"4.The beverage apparatus 30, as will be described in greater detail herein below, relates to a beverage brewing or producing system whereas the soluble beverage apparatus 30a, as will be described in greater detail herein below, relates to a soluble or powdered beverage dispensing system.\",\n \"Both beverage apparatuses 30, 30a employ a common heated water reservoir contained in housing 34 to which the individual modules 32, 32a are connected.\",\n \"As shown in FIGS.\",\n \"2-4, the housing 34 desirably is centrally located to serve multiple beverage modules 32, 32a.\",\n \"The benefit of a common housing 34 functioning as a common reservoir to supply water, instead of individual tanks for each module 32, 32a, is that it substantially reduces the cost and greatly increases the efficiency of the system.\",\n \"Also it reduces the redundancy of multiple heated water reservoirs, reduces the wattage required by the system and increases flexibility in the number of modules 32, 32a which might be included.\",\n \"The particular configuration with the housing 34 positioned in close proximity to or between several modules 32, 32a is beneficial in that it also minimizes the length of water lines 38 connecting the housing 34 to the modules 32, 32a.\",\n \"Additionally, a larger body of water in the housing 34 tends to help maintain the temperature and reduce temperature loss over time as well as maintaining a larger volume of brew water at a desired temperature.\",\n \"While the housing 34 retained within the beverage apparatus 30 is insulated to reduce heat loss and reduce heat load on the ambient environment, heat which may escape from the housing 34 will generally pass to the modules 32, thereby increasing the efficiency of maintaining the temperature of the beverages retained in the modules 32.Insulation desirably is provided to reduce heat loss to the ambient environment.\",\n \"This will also help reduce the exertion on the ambient air-conditioning system thereby further increasing efficiency of the present beverage apparatus 30, 30a.\",\n \"With reference to FIGS.\",\n \"1-4, each module 32, 32a produces a separate brewed beverage.\",\n \"The centralization of a housing 34 helps reduce the individual cost associated with the module 32,32a and improves the ease of manufacturing of the modules.\",\n \"Also, there is increased commonality of parts associated with this type of system.\",\n \"The modular configuration of the beverage apparatus 30 using the modules 32, 32a increases the potential variability of the physical size and brewer configuration of the system.\",\n \"This is beneficial to accommodate the needs of a particular location.\",\n \"In other words, only a desired or required number of modules 32, 32a will be needed at any given time.\",\n \"If a location has an increase in demand for brewed or soluble beverages additional modules 32, 32a may be added as necessary for the particular change in demand.\",\n \"Additionally, with regard to repair and maintenance, the beverage apparatus 30, 30a can be designed such that individual modules 32, 32a can be removed for repair and maintenance as necessary with an identical module being inserted in the missing modules.\",\n \"In a preferred embodiment, (FIG.\",\n \"5) the housing 34 includes a heating element 52, and one or more system sensors such as a level sensor 86 and a temperature sensor 84.Such components tend to be generally readily available and easily replaced and as such further enhance the reliability of the overall beverage apparatus 30, 30a.\",\n \"With reference to FIG.\",\n \"3, the line 38 connecting each housing 34 to a brewing portion 40 of the module 32 is shown.\",\n \"Water 42 retained in the housing 34 can be readily dispensed.\",\n \"A water inlet line 44 is provided to introduce unheated water 50 to the housing 34.The inlet line 44 runs from an upper portion 46 of the housing 34 and directs the inward water downwardly towards a bottom portion 48 of the housing 34.Introducing the unheated water 50 towards the bottom of the reservoir introduces the unheated water 50 in close relation to a heated element 52 thereby accelerating the heating of the water.\",\n \"Additionally, the introduction of unheated water 50 in the bottom portion 48 tends to force heated water in the upper portion 46 upwardly.\",\n \"As such, the water 42 in the upper portion 46 tends to be the hottest water in the housing 34.Introducing unheated water 50 from the upper portion has the benefit of providing an entrance into the reservoir through a reservoir wall 56 with little or no pressure on the entry therethrough.\",\n \"This may result in an increase in the reliability of the entry therethrough.\",\n \"Additionally, a reinforcing or strengthening structure 58 is provided extending between opposed walls 60, 62 of the housing 34.This helps prevent bowing or stress on the walls 60, 62 which might otherwise result from the force of the water on the walls.\",\n \"Multiple reinforcing structures 58 may be provided throughout the housing 34 as necessary to provide support for the size and dimension of the housing required for a particular system.\",\n \"FIG.\",\n \"5, for example, shows a diagrammatic illustration of an embodiment of the apparatus 30 shown in FIGS.\",\n \"1-4.The beverage dispensing modules 32 can have any suitable construction desirably adapted to produce beverages, store beverages for ready consumption, and facilitate ready dispensing of the beverages.\",\n \"Each module 32 includes a brewing assembly 30 that may have any suitable construction that can be used to produce beverage such as brewing coffee or other beverages.\",\n \"The illustrated brewing assembly 30, for example, includes a brewing substance retainer or assembly 64 including a funnel 66.Such a retainer 64 may be well known in the art and may include a filter (not shown) retained in the funnel 66 for holding a charge of beverage substance.\",\n \"The funnel 66 may be positioned above a reservoir or container portion 68 of the dispensing module 32 which receives brewed beverage from the funnel.\",\n \"A dispensing assembly 70 in the form of a controllable valve may be associated with the container 68 for dispensing beverage from the module 32.With reference to FIG.\",\n \"5 a simplified review of the beverage brewing process is described.\",\n \"A brew actuator switch 72 desirably is associated with each of the modules 32.To initiate a brew cycle for one of the modules 32 a respective brew activation switch 72 is operated.\",\n \"Operation of the switch 72 sends a signal over line 74 to controller 76.Controller 76 operates an inlet valve 78 over line 80 to introduce water to the housing 34.Also, heating element 52 is coupled to the controller 76 over line 82.The temperature sensor 84 and level sensor 86 are coupled to the controller over lines 88, 90 respectively.\",\n \"If the controller 76 detects proper level and temperature it will initiate a brew cycle thereby displacing water from the housing 34 through a sprayhead assembly 92.The sprayhead assembly 92 communicates with the substance retainer 64 and dispenses water into the funnel 66 for producing a beverage therein.\",\n \"The illustrated module 32 includes a level sensor 94 associated with the container 68 which is coupled to the control over line 96.The level sensor can detect a change in level in beverage retained in the container 68 which may be used to activate a brew cycle.\",\n \"Also, the level detector 94 may sense the level in the beverage container 68 thereby indicating if the container is full or above the desired level to prevent initiation of a brew cycle.\",\n \"This will prevent over flow of the beverage server 68.The beverage dispensing modules 32 can be constructed to dispense beverage in any suitable manner.\",\n \"In a preferred embodiment, for example, a dispenser or faucet switch 98 may be associated with the dispensing assembly 70 and coupled with a dispensing handle or control 100.Activation of the control 100 will activate the switch 98 over line 102 connected to the controller 76 thereby indicating that the controller 76 should operate a solenoid valve 104 (FIGS.\",\n \"6 and 7), via line 106 to dispense beverage therefrom.\",\n \"As will be described in greater detail below with regard to FIGS.\",\n \"6 and 7, a disposal or dump valve 108 is also coupled with the container 68 to facilitate controllable disposal, via line 110 coupled to the controller 76.With reference to FIGS.\",\n \"6 and 7, the beverage container 68 may further include a manifold 114.The illustrated manifold 114 includes a primary passage 116 which communicates with an interior cavity or reservoir 118 of the container 68.Beverage flows from the cavity 118 through the primary passage 116.The solenoid valves 104, 108 communicate with the passage 116.A dispensing passage 120 and a disposal passage 122 communicate with the primary passage 116.Solenoid valve 104 and 108 desirably are normally closed and require specific activation in order to open the valves.\",\n \"During a dispense cycle the switch or control 100, for example a controllable faucet as shown in FIG.\",\n \"5, is activated thereby activating solenoid valve 104.When the dispense solenoid valve 104 is opened beverage 124 will flow from the cavity through the solenoid valve 104.Under this condition disposal valve 108 is closed.\",\n \"When beverage must be disposed of, the disposal solenoid valve 108 is opened while maintaining the dispense solenoid valve 104 in a closed position.\",\n \"This will allow the disposal of beverage through a separate dispensing line.\",\n \"Disposal of beverage may occur as a result of a time lapsed period whereby after a specified period of time beverage should be disposed of.\",\n \"For example, if a beverage only maintains a desired flavor profile or other characteristics for a period of time, for example 2 hours, at the end of two hours after the beverage brewing cycle is initiated, the beverage may be disposed of through the disposal valve 108.Such disposal may also be used at the end of a service cycle such that at the end of the day all beverages may be disposed of at a predetermined time.\",\n \"The disposal valve may also be used in a manner to flush and cleanse the beverage brewing system.\",\n \"In this regard, at a predetermined period of time, the controller 76 may operate the disposal valve 108 to dispose of any beverage retained in the container 68.The disposal valve 108 and dispense valve 104 may be closed.\",\n \"The controller 76 may then dispense a quantity of water into the cavity 118 thereby flushing and sanitizing the reservoir.\",\n \"After holding the heated water for a predetermined period of time the water may be disposed through the disposal valve 108.Multiple rinsing or flushing cycles may be controlled by the controller 76.In this regard, both the cavity 118, the container 68 and the associated passages 116, 122, 120 and solenoid valve 104, 108 maybe cleansed and sanitize periodically and automatically.\",\n \"It will be appreciated that these automatic cycles will likely greatly increase the cleanliness, taste reliability and sanitation of such beverage dispensing systems as such procedures require considerable amount of time on the part of manual operators.\",\n \"The use of controllable solenoid valves 104, 108 also prevents dispensing of beverage during a brew cycle.\",\n \"For example, once a brew cycle is initiated the controller 76 can lock out the solenoid valves 104, 108.This will allow for the brewing cycle to complete and brewed beverage to mix within the container 68 to insure consistent flavor.\",\n \"The beverage dispensing modules 32 can be constructed in any other suitable manner to dispense brewed beverage in any suitable manner in accordance with various embodiments disclosed.\",\n \"If desired, for example, the beverage containers 68 of the beverage dispensing modules 32 can instead be in the form of carafes (not shown) or have any other suitable construction.\",\n \"In a preferred embodiment, during a brewing cycle, heated water 134 is dispensed from the housing 34 through the water dispensing system.\",\n \"Heated water 134 is used to infuse a beverage brewing substance 136 retained in the brewing assembly 64.An opening 138 is provided in the bottom of the funnel 66 to allow a brewed beverage 140 to flow therefrom.\",\n \"As shown, for example, in previous figures, a brewed beverage container 68 is provided to collect the beverage 140.The beverage container 68 may be of any suitable form.\",\n \"With reference to FIG.\",\n \"8, a liquid flavor dispensing system 130 may be employed with the modular beverage dispensing system disclosed.\",\n \"The liquid flavor dispensing system 130 can have any suitable configuration and can be used to supply flavoring 132 to all or less than all of the beverage dispensing modules 32.The liquid dispensing system 130 may also be used to supply a different flavoring 132 to each of the different beverage dispensing modules 32.Additionally, the flavorings 132 can be supplied to the beverage dispensing modules by a single liquid flavor dispensing system 130 or several liquid flavor dispensing systems.\",\n \"Further, the flavoring 132 may in the form of the typical flavorings associated with brewed coffee or instead be in any other suitable form such as, for example, honey, peach, lemon, herbal flavors, or any other suitable flavoring which might be desired by a beverage consumer for any number of different beverages.\",\n \"In a preferred embodiment, a small quantity of flavoring 132 is introduced into the water which is dispensed to the beverage brewing substance 136.The beverage brewing substance may be in the form of decaff or regular ground coffee as well as tea substances or other substances which might be used in an infusion beverage brewing process.\",\n \"The objective in using the liquid flavoring material 132 is to introduce a small quantity of concentrated material or substance 132 during the brewing process so that the flavor becomes fully mixed with and saturated in the heated water 134.Introduction of the flavor 132 early in the brewing process will help assure that the flavor fully mixes with the water, is fully flushed through the beverage brewing substance to maximize the value of the flavor and is used to produce an aroma which might entice purchasers of the beverage as part of the beverage purchasing experience.\",\n \"The flavor could also be dispensed into the beverage brewing substance and subsequently mixed with the water and the resultant brew during the brewing process.\",\n \"A benefit of adding flavors at the time of brewing and/or mixing soluble beverages, over using pre-flavor beverage substances, is the increased effectiveness of the flavorings.\",\n \"The increased effectiveness of the flavorings reduces the amount of flavoring required to produce a desired taste profile.\",\n \"Under these circumstances, the flavorings are mixed with the beverage substance under optimal conditions to maximize the flavoring benefits of the flavorings.\",\n \"For example, the time prior to mixing the flavoring with the brewing substance is nominal thereby virtually eliminating any degradation of the flavoring which might occur if the flavoring was exposed to air.\",\n \"A liquid flavor dispensing system 130 is illustrated diagrammatically in FIG.\",\n \"8.With this embodiment, the liquid flavor dispensing system dispenses a different liquid flavoring 132 in a controlled manner to several of the brewing assemblies of the beverage dispensing modules 32.The flavor can be dispensed to a water or liquid distribution system which controllably dispenses water to a beverage making substance, the beverage malting substance or both.\",\n \"With reference to FIGS.\",\n \"8-10, the illustrated liquid flavor dispensing system 130 desirably includes a plurality of packages or containers 144a-144h, each of which retains a respective liquid flavor 132 therein.\",\n \"The containers 144 desirably are in controlled communication with respective beverage dispensing modules 32a-32d in any suitable manner.\",\n \"For example, a section of tubing or other passage 146 may connect each container 144 to a respective check valve 148.The check valve 148 is part of a respective solenoid or other form of pump device 150 coupled to the controller 76 via a respective line 152.A respective small diameter tube 154 extends from the solenoid pump 150 to a respective second check valve or flavor injection port 156 at a respective terminal end 158 of the tube 154.A respective sensor 160 can be coupled to the controller via a respective line 162 to monitor and detect a low level of the respective flavoring 132 in the respective container 144.As will be described in greater detail below with regard to sensing and monitoring, the status of the flavorings 132 can be reported by the controller 176.The check valves 148 and 156 at each end of each of the tubes 154 provide a sealed line to which the container 144 can be attached.\",\n \"Also, either alternatively or in addition to the use of the sensor 160, the flavor usage can be inferred based on information collected on a characteristic of the pump such as operation or usage for example, pump actuation.\",\n \"For example, a sensor may be associated with the pump and run time of the pump 150 could be monitored and a calculation made to infer the amount of flavorings remaining in a corresponding flavorings container 144.The system would provide a signal to the operator, such as “checlc-status” when the level of flavorings approaches, attains or drops below a predetermined level, for example 20% remaining.\",\n \"Such advance warning would give the operator the opportunity to change the flavorings container 144 before it runs out.\",\n \"The advance warning also helps assure that a reasonable margin of error in the inferred calculation is considered and accommodated for.\",\n \"In use, when a brewing cycle is initiated, the solenoid pump 150 can be activated simultaneous with the initiation of the brewing cycle of one of the beverage dispensing modules or shortly thereafter.\",\n \"A quantity, for example 9 ml for use in brewing ½ gallon of coffee, of liquid flavoring 132 is pumped from the container 144 through the tubing 154.The check valve or flavor injection port 156 is positioned in close proximity to a sprayhead 170 of the dispensing assembly 92.Dispensing of the flavor 132 into the water will allow the flavor to mix with the heated water for brewing substance during the saturation and extraction process.\",\n \"Generally, it is desirable to use the flavoring at the start of the cycle which will result in completely integrating the flavor during the brewing cycle.\",\n \"The process may be repeated with other beverage dispensing modules 32, 32a and, if desired, with other flavors.\",\n \"The flavor injection ports 156 may, for example, include a check valve of a type as produced by Smart Products.\",\n \"This is a positive pressure inverted check valve which requires positive pressure through the tube 154 to open the valve.\",\n \"Positioning of the injection port 156 in close proximity to the sprayhead 170 results in the benefit of clearing and rinsing the surface as a result of the water vapor and heat during a brewing cycle.\",\n \"Periodic manual cleaning of the sprayhead 170 and injection port 156 helps maintain a clean system.\",\n \"Desirably, because the injection port 156 is an inverted pressure check valve, physical contact with the tip of the injection port 156 through which the liquid 132 is dispensed does not result in dispensing of liquid 132.Rather, physical contact with the end of the injection port 156 causes the valve to close as opposed to open.\",\n \"The system also prevents the entry of atmosphere into the container 144 or line 154 thereby preventing contamination or degrading of the flavor or quality of the flavorings.\",\n \"This system provides a generally closed system which prevents the flavorings contact with the atmosphere and prevents contamination providing a generally aseptic environment.\",\n \"With reference to FIG.\",\n \"11, the liquid flavor dispensing system 130 as described with regard to FIGS.\",\n \"8-10 may be used with a soluble beverage system 176.In FIG.\",\n \"11, the liquid dispensing system dispenses liquid flavor into a hot water path 174 of a soluble or powdered beverage dispensing system 176.The devices used to dispense the liquid flavor are the same as those describe above with regard to FIG.\",\n \"8.As such, only the terminal ends 158a, 158b and 158c of multiple lines 154a, 154b, and 154c are shown.\",\n \"Similarly, the multiple injection ports 156a, 156b, and 156c are shown.\",\n \"The soluble beverage dispensing system 176 retains a quantity of powdered beverage mix (not shown) in a beverage dispensing container 178.The dispensing of soluble beverage material in powder form is generally known in the art.\",\n \"The container 178 includes a dispensing auger (not shown) which controllably dispenses powder by operation of a dispensing drive or motor 180 controlled by the controller 76a.\",\n \"Powdered beverage material is dispensed through the passage 182 into a mixing chamber 184.The hot water line 174 communicates with the mixing chamber 184 to combine with the powder.\",\n \"The combined powder and hot water flow into an agitating chamber 186 there below.\",\n \"A whipper motor 188 and agitating blade 190 further blend the powder and water combination for dispensing a beverage 192 through a nozzle 194.The flavor dispensing system 130 (see, FIG.\",\n \"8) controllably dispenses liquid flavor 132 from respective containers 144 using corresponding pumps via the lines 154a, 154b, and 154c.\",\n \"Either individual flavors or combinations of flavors may be dispensed into the hot water line.\",\n \"Such dispensing may be programmed in the controller 76a.\",\n \"Positioning of the injection ports 156a, 156b, 156c in the heated water path helps to assure that the liquid flavors are rinsed from the injection ports during and at the conclusion of the heated water dispensing cycle during a beverage dispensing cycle.\",\n \"It should be noted that generally these powdered beverage dispensing systems have a momentary hot water overrun at the end of the cycle during which powdered beverages dispense from the container 178.The slight overrun of heated water helps to assure that the mixing chamber 184, whipping chamber 186, and nozzle 194 are rinsed clean at the end of the cycle.\",\n \"The injection ports 156a, 156b and 156c have been positioned in the hot water line to provide the rinsing characteristics and benefits of the overrun of hot water.\",\n \"Furthermore, it is commonly known to have at least one daily rinse cycle in which heated water is dispensed through the assembly 176 to maintain cleanliness and sanitation of the system.\",\n \"During a brew cycle as shown in FIG.\",\n \"8, liquid flavor is generally preferably dispensed at the initiation of the brew cycle.\",\n \"As noted, this helps to ensure complete mixing and optimal extraction of the flavor from the beverage brewing substance 136.A predetermined volume of liquid flavor 132 can be dispensed into the brewing assembly 30 at the initiation of the brew cycle.\",\n \"In contrast, the soluble system 176 as shown in FIG.\",\n \"11 may have a variable quantity of beverage 192 dispensed therefrom.\",\n \"It is anticipated that multiple beverage sizes may be dispensed from the soluble system 176.As a result, the liquid flavoring 132 must be dispensed at a rate consistent with the concentration flavor profile or recipe, and powder also dispensed by the system.\",\n \"For example, if a consumer wishes to obtain a 12 ounce beverage the proper amount of liquid flavoring must be dispensed for a 12 ounce beverage.\",\n \"However, the consumer may be allowed to control whether they wish to have a full 12 ounces of beverage or if they decide to short the beverage resulting in, perhaps, only 10 ounces being dispensed.\",\n \"As such, the pump associated with the powdered beverage system 176 must be calibrated to dispense small quantities of liquid flavoring 132 on a per unit time basis to assure a desirable and consistent mixing of flavor with the powder.\",\n \"This will help assure that the desired flavor is achieved regardless of the quantity of beverage dispensed.\",\n \"The solenoid pump 150 used with the liquid flavor dispensing system 130 is a precision controlled metering pump.\",\n \"It is anticipated that other forms of the metering pump may be used with this system.\",\n \"However, in the present embodiment, the metering pump is controllably operated to dispense very small quantities of the liquid flavor 132.The frequency of the plunger rate, the stroke of the plunger as well as the duration of plunger operation the solenoid pump 150 can be controlled by the controller 76, 76a.\",\n \"As a result, very small quantities of highly concentrated liquid flavor can be dispensed into the beverages 140, 192 during the corresponding beverage preparation process.\",\n \"As such, precise quantities can be added to provide flexibility and tailoring of the resultant beverages.\",\n \"The system 130 can be calibrated using volumetric guidelines associated with various beverage recipes and flavor materials to match particular flavor preferences, for example for various demographics.\",\n \"As such, a franchise operation can custom blend and configure the flavor of beverages dispensed by the beverage dispensing system to meet and satisfy regional preferences.\",\n \"Additionally, if preferences change over time the system can be modified and calibrated to match those preferences.\",\n \"This adds further flexibility to the beverage system disclosed.\",\n \"With reference to FIGS.\",\n \"14-18, a powdered or soluble beverage dispensing system 199 is shown.\",\n \"While a soluable beverage system 199 has been shown and described as a powder dispensing system, it is envisioned that such a soluable system also includes a liquid concentrate system which dispenses a quality of liquid coffee concentrate for mixing with water or other dilution or reconsituting substance.\",\n \"Reference to a soluable beverage system is intended to be broadly interpreted.\",\n \"Such a beverage dispensing system has been generally and diagrammatically shown in FIG.\",\n \"11.With further reference to FIG.\",\n \"14, the container 178 is shown positioned inside a housing 200.The passage 182 coupled to the container 178 is shown positioned for dispensing into the mixing chamber 184 and communicating with the whipping chamber 186 and nozzle 194.A portion of the housing 200 in the form of a door 202 conceals the powdered beverage system 176 in the housing 200.Also, the liquid flavor dispensing system 130 as described hereinabove may be used with the powdered beverage or soluble beverage dispensing system 176 illustrated in FIGS.\",\n \"14-18.One of the potential difficulties involved with a soluble beverage dispensing system is that the container 178 must be removed periodically for refilling with powdered or soluble beverage substance as well as regular maintenance and cleaning.\",\n \"The container 178 may not be very heavy when it is empty because it is generally formed of a plastic material.\",\n \"However, once filled, it may contain 5 to 10 pounds of powder material.\",\n \"With the container 178 being positioned in an elevated location and containing a quantity of material, it may be difficult to lift the container into a desired position in the housing 200.The container 178 is positioned in an upper position in the housing 200 in order to benefit from gravity assistance when dispensing powdered beverage therefrom to the mixing chamber 84.Additionally, it is important to align the dispensing passage 182 with the housing 184 to help assure that when powdered beverage material is dispensed from the container 178 it flows into the proper path for mixing with water.\",\n \"As such, it would be desirable to provide an apparatus and system for facilitating improved ease of removal and replacement of the powdered beverage container 178 relative to the housing 200.As shown in FIGS.\",\n \"14-18, a container positioning assembly or container positioner 220 in the form of a slide-out support or shelf is provided.\",\n \"The container positioning assembly 220 includes a pair of handles 222 which can be grasped (see FIG.\",\n \"15) by a user to extract the container 178 attached to the assembly 220 for removing it from the housing 200.As shown in FIG.\",\n \"15, the assembly 220 allows the container 178 to be pulled straight out of the housing 200 thereby allowing disengagement of the passage 182 relative to the mixing chamber 184.Movement of the assembly 220 also replaces the container 178 in a desired position to align the dispensing passage 182 with the mixing chamber 184.As can be seen in FIGS.\",\n \"15-17, the container 178 has been removed from the housing 200 in order to refill the container 178 but does not have to be removed from the shelf 250 of the assembly 220.Support rails 226 are positioned on each side of container 178 and are engaged with side walls 228 of the housing 200 (FIG.\",\n \"15).\",\n \"Each of the guide rails 226 includes a pair of tracking slots 230, 232 which are engaged with corresponding upper 234 and lower 236 guide rollers.\",\n \"A support roller 240 is positioned along an exterior edge 242 of the guide 226 to further help facilitate smooth and controlled movement of the assembly 220.Support rollers 240 and guide rollers 234, 236 are mounted on the sidewalls of the housing 200.The guides 226 are connected by a cross extending shelf 250 on which the container 178 is positioned.\",\n \"The combination of the guide rails 226 and rollers help the assembly 220 slide outwardly and inwardly relative to the housing 200.FIG.\",\n \"16 shows only the portion of the guide rails 226 extending past the edge of container 178.The guide rollers 240, 234, 236 associated with the guides 226 prevent tipping of the shelf 250 and associated container 178.When the assembly 220 is stowed in the housing 200 the shelf 250 rests on a corresponding surface 252 of the housing 200.As can be seen in FIGS.\",\n \"15-17, the guides 226 include a generally horizontal portion 254 and a vertically oriented portion 256.The shelf 250 and container 178 are associated with the generally horizontal portion 254.The generally vertical portion 256 helps to allow forward movement as well as downward movement of the assembly 220 relative to the housing 200.The guide including the portion 254, 256 help facilitate movement of the container into and out of the housing 200 and forward and downward movement.\",\n \"In the embodiment as shown, the assembly facilitates positioning of the container 178 approximately 6-8 inches downwardly from its stored position in the housing 200.This lowering of the container 178 is beneficial in that it allows for easy filling of the container.\",\n \"Also, by fixing the container to the shelf 250 and the assembly 200 it prevents spilling or otherwise tipping of the container 178 during filling or other processes.\",\n \"This can result in savings in terms of lost beverage material as well as cleanup costs and associated complications.\",\n \"In the stored position, detents 260, 262 are provided on the guides 230, 232, respectively.\",\n \"The detents 260, 262 allow the rollers 234, 236 respectively, to be engaged therewith to prevent casual disengagement of the assembly 220 from the housing 200.This helps to lock or retain the assembly 220 in a desired stored or stowed position in the housing 200.As the assembly 220 is moved outwardly from the housing 200, the track guides 234, 236 track along the corresponding channels 230, 232 generally horizontally along the horizontal portion 254.As such, the container 278 attached to the shelf 250 sides forwardly and outwardly from the housing.\",\n \"At a position where the generally horizontal portion 254 transitions into the partially vertical portion 256 (elbow 270) the guides allow the assembly to slide downwardly and forwardly.\",\n \"In the extended most position the guides 234, 236 engage corresponding extended detents 272, 274, respectively.\",\n \"These detent positions 272, 274 help retain the assembly 220 in the outward or extended most position.\",\n \"The assembly 220 also includes an assist system 280 which includes cables 282 and biasing members to springs 284 forming a cable-biasing assembly help facilitate removal of and replacement of the container 178 relative to the housing 200.With reference to FIG.\",\n \"18, the assist system 280 includes a spring 284 attached at one 286 to a back portion of the housing and at a second end to a first pulley 288.The cable 282 is attached at a first end 290 to a corresponding portion of the housing and extends through the first pulley 288 and up and around a second pulley 292.The second end 294 of the cable 282 attaches to a corresponding portion of the guides 226 at attachment point 296.Attachment of the cable 282 to the guide 226 is shown in FIG.\",\n \"16.The assist assembly 288 helps further facilitate ease of removal and replacement of the container 178 relative to the housing 200.For example, when the container 178 is removed from the housing on the assembly 220 the spring 284 stores energy and is stretched as the cables 282 are extended.\",\n \"The cables 282 are extended as a result of the forward and downward movement of the guides 226.Extension of the springs 284 expends the stored energy and imparts a return force in the springs which helps reduce the force required to return the container 178 to the housing 200.This is particularly useful when an empty container is removed from the housing, filled with powdered material and then returned.\",\n \"Returning the filled container 178 normally would require additional effort on the part of the operator and as such the return forces in the assist system 280 help make the return easier.\",\n \"Turning now to FIGS.\",\n \"19 and 20, the beverage system also includes a communications system or means 300 for communicating 302 between a corresponding beverage brewing, beverage making apparatus or beverage dispensing modules 32, 32a and a receiver or base station 304.Each of the modules 32, 32a includes a controller 76, 76a which is centralized for use with multiple modules or dedicated to an individual module.\",\n \"The controller collects information about the module or modules.\",\n \"For purposes of this present discussion, we will refer to the controller as being dedicated to multiple modules and receiving information from each individual module associated therewith.\",\n \"The many modules associated with a single control also provides the added benefit of the modular assembly as described hereinabove.\",\n \"It should also be noted that the details of the circuitry used to achieve the communication system are illustrated in the detailed schematics provided in FIGS.\",\n \"21-28 and all corresponding sub-portions thereof.\",\n \"For example, FIG.\",\n \"21 is a general schematic of the overall system.\",\n \"FIGS.\",\n \"22-28 provide details about the general system shown in FIG.\",\n \"21.FIG.\",\n \"21 has been noted with cross reference to the other figures which provide details about the portions of the circuitry shown generally in FIG.\",\n \"21.The controllers 76, 76a include the link 302 which connects the controller to the corresponding receiver or base station 304.In the present embodiment as diagrammatically illustrated in FIGS.\",\n \"19 and 20, the controllers 76, 76a communicate with the receiver 304 by way of an RF link 302.Generally, the controllers are remote from the receiver and use the Link 302 and the associated means for linking to communicate the collected information about the beverage making apparatus.\",\n \"Antennae 306, 308 are provided on the controllers 76, 76a and the corresponding receivers 304 respectively.\",\n \"It should be noted that the communication link 302 may be in any one of a variety of forms such as hardwired physical point-to-point link, optical lines, light wave, ultrasonic, infrared or any other form of communication link between one or more devices.\",\n \"The receiver 304 receives information from the controller or various components of the modules 32, 32a to identify information relating to the modules and operation of the modules.\",\n \"The receiver 304 may be in the form of a monitoring unit positioned in an appropriate location relative to an operator or attendant who has responsibility for or otherwise maintains the modules 32, 32a.\",\n \"The receiver 304 includes a means for displaying display 305 and can provide visual auditor or other information about numerous conditions.\",\n \"The display 304 allows a form of the collected information about the beverage making apparatus to be displayed at the receiver.\",\n \"The visual display may include, but is not limited to lights, text, symbolic images (i.e., dispenser “full” showing a colored or shaded dispenser and a dispenser “empty” showing an unshaded dispenser), and mechanical devices that tare operated or other visual displays.\",\n \"An auditory display may include, but is not limited to speech information, alarms, tones or other signals that van be heard.\",\n \"Tactile displays are also contemplated in the form of a vibrating surface, vibrating device worn by the operator or other means for letting the operator or other means for letting the operator know about he collected information.\",\n \"For example, the following is a list of the messages or conditions which can be displayed: Receiver fault; Please wait attempting communication; Communication failure, coffee module #; Communication fault, soluble module #; Flavor [type] low, coffee module #; Flavor [type] low, soluble module #; Hopper low, soluble module #; Hopper empty, soluble module #; Coffee empty, coffee module #; Coffee low, coffee module #; Freshness expired, coffee module #; Dumping in ______ minutes, coffee unit #; Server removed, coffee module #; Refill too long; Temperature probe open; Temperature probe short; Heating too long; Over flow safety; Coffee stations all OK; and Soluble stations all OK For example, in an convenience store setting, multiple modules 32, 32a may be generally remotely positioned in one portion of the store, for example, a rearward location of the store, if desired.\",\n \"The operator of the beverage system may also be the same person responsible for stocking of the equipment, and register activities.\",\n \"As such, one person is required to operate and maintain many components of the convenience store facility.\",\n \"The receiver 304 may be positioned at the cash register so that while the attendant is operating the cash register he may also be notified of matters that require his attention at the modules 32, 32a.\",\n \"For example, if a module 32a indicates that a container 68 is empty the controller 76 will communicate with the receiver 304 to indicate this condition.\",\n \"The receiver may also be configured with audible alarms and visual displays to provide additional information.\",\n \"For example, a display may identify which module and the specific condition associated with the module which has provided a signal to the receiver 304.The operator can then make a decision how to use that information relative to his responsibilities at the cash register.\",\n \"If an emergency requires immediate attention he can secure the cash register and address the emergency while attending to the module 32.Alternatively, if it is a slow period in the store and a module is indicating that a container 68 is empty he can put off attending to this condition until he completes one or more transactions in queue.\",\n \"The signal communication between the controller 76 and the receiver 304 may be a single path signal or in the form of a multiple path verification signal.\",\n \"For example, as shown in FIGS.\",\n \"19 and 20 a first signal 310 is transmitted from the controller 76 to the receiver 304.This initial communication may report the status of a condition at the module 32.The receiver 304 can then return a copy of the information 312 to the controller 76.If the controller confirms the information received (312) it can then send a return confirmation 314 that the information is correct.\",\n \"This is a useful communication link in the present situation due to the presence of electrical noise and other interference within other beverage system applications.\",\n \"For example, each of the components associated with the modules 32, 32a may provide some degree of electrical or other noise.\",\n \"Additionally, other systems within the beverage system setting may be producing RF or other signals which could create interference.\",\n \"As such, the present embodiment providing the verification communication links 310-314 helps facilitate and assure accurate and timely communication.\",\n \"In the event of excessive interference such as from patrons of the store using telephone or RF communication devices, the controller 76 can continue to periodically send a message 310 to the receiver 304 until the verification signal 312 is received.\",\n \"While preferred embodiments are disclosed, illustrated and described, it is envisioned that those skilled in the art may devise various modifications and equivalents without departing from the spirit and scope of the disclosure as recited in the following claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10477002"}}},{"rowIdx":993,"cells":{"text":{"kind":"list like","value":[["Methods and compositions for prevention of angioproliferation","The invention provides pharmaceutical compositions comprising Porphyromonas gingivalis protease and hemagglutinin polypeptides that have anti-angiogenic activity and methods for their use."],["1.A composition for the treatment, prevention, or amelioration of an angioproliferitive condition comprising a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide has anti-angiogenic activity, and a pharmaceutically acceptable excipient.","2.The composition of claim.","1, wherein the polypeptide is selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.","3.The composition of claim 2, wherein the polypeptide has at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, prtT or tla polypeptide and wherein the polypeptide has anti-angiogenic activity.","4.The composition of claim 2, wherein the polypeptide is a biologically functional homolog, or isoform of a rgpA, rgpB, kgp, hag, prtT or tla polypeptide.","5.The composition of claim 1, wherein the polypeptide is a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or Porphyromonas gingivalis hemagglutinin polypeptide, and wherein the fragment has anti-angiogenic activity.","6.The composition of claim 5, wherein the fragment is HA2.7.The composition of claim 1, wherein the angioproliferative condition is carcinoma, sarcoma, melanoma, benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthris, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, arteriosclerosis, scleroderma, hypertrophic scars, cat scratch disease, Helicobacter pylori ulcers, capillary proliferation within atherosclerotic plaque, or a combination thereof.","8.A method for the treatment, prevention, or amelioration of an angioproliferative condition comprising administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof wherein the polypeptide has anti-angiogenic activity, to a patient in need thereof whereby the angioproliferative condition is treated, prevented or ameliorated.","9.The method of claim 8, wherein the polypeptide is selected from the group co-ng of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.","10.The method of claim 8, wherein the polypeptide has at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, or tla polypeptide and wherein the polypeptide has anti-angiogenic activity.","11.The method of claim 8, wherein the polypeptide is a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or a Porphyromonas gingivalis hemagglutinin polypeptide, ad wherein the fragment has anti-angiogenic activity.","12.The method of claim 11, wherein the polypeptide fragment is HA2.13.The method of claim 8, wherein the angioproliferative condition is carcinoma, sarcoma, melanoma, benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthritis, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, atherosclerosis, scleroderma, hypertrophic scars, cat scratch disease, Helicobacter pylori ulcers, capillary proliferation within atherosclerotic plaque, or a combination thereof.","14.The method of claim 8, wherein the method comprises contacting a vasculature supplying a biological structure affected by the angioproliferative condition with the polypeptide.","15.The method of claim 14, wherein the polypeptide is contacted with a basolateral surface of the vasculature.","16.A method for potentiating effects of a chemotherapeutically effective agent comprising administering to a patient: (a) a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptid, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide disrupts cell-cell adhesion, cell-matrix adhesion, or both; and (b) a chemotherapeutically effective agent, whereby effects of a chemotherapeutically effective agent are potentiated.","17.The method of claim 16, wherein the polypeptide is selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.","18.The method of claim 17, wherein the polypeptide has at least about 900% sequence identity with a rgpA, rgpB, kgp, hag, or tla polypeptide, and wherein the polypeptide has anti-angiogenic activity.","19.The method of claim 16, wherein the polypeptide is a fragment of Porphyromonas gingivalis arginine specific cystine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or Porphyromonas gingivalis hemagglutinin polypeptide, and wherein the fragment has proteinase activity.","20.The composition of claim 19, wherein the polypeptide fragment is HA2.21.A method for preventing the formation of new vasculature required for implantation or sustenance of a fertilized an ovum comprising: administering a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a mammal, wherein the polypeptide has anti-angiogenic activity, whereby the formation of new vasculature required for implantation or sustenance of a fertilized mammalian ovum is prevented.","22.The method of claim 21, wherein the polypeptide is selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.","23.The method of claim 22, wherein the polypeptide has at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, prtT, or ta polypeptide and wherein the polypeptide has anti-angiogenic activity.","24.The method of claim 21, wherein the polypeptide is a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or Porphyromonas gingivalis hemagglutinin polypeptide, and wherein the fragment has anti-angiogenic activity.","25.The method of claim 24, wherein the polypeptide fragment is HA2.26.A pharmaceutical composition for facilitating passage of a compound through a blood-brain barrier comprising a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, a Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, and a pharmaceutically acceptable excipient.","27.The pharmaceutical composition of claim 26 further including a compound to be passed through the blood-brain barrier.","28.A method delivering a compound through a blood-brain barrier of a patient comprising administering a pharmaceutically effective amount of a Porphymonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof and the compound, whereby the compound is passed through the blood-brain barrier.","29.The method of claim 28, wherein the polypeptide is selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.","30.The method of claim 29, wherein the polypeptide has at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, prtT or tla polypeptide and wherein the polypeptide has anti-angiogenic activity.","31.The method of claim 28, wherein the polypeptide is a foment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or a Porphyromonas gingivalis hemagglutinin polypeptide, and wherein the fragment has anti-angiogenic activity.","32.The method of claim 31, wherein the polypeptide fragment is HA2.33.A method of degrading a tumor comprising: administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a patient having a tumor, wherein the polypeptide degrades cell-cell bonds, cell-matrix bonds, or both cell-cell bonds and cell-matrix bonds of the tumor, whereby the tumor is degraded.","34.The method of claim 33 wherein the polypeptide is selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.","35.The method of claim 34 wherein the polypeptide has at least about 90% se ce identity with a rgpA, rgpB, kgp, hag, prtT or tla polypeptide and wherein the polypeptide has anti-angiogenic activity.","36.The method of claim 33 wherein the polypeptide is a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or Porphyromonas gingivalis is hemagglutinin polypeptide, and wherein the fragment has anti-angiogenic activity.","37.The method of claim 36 wherein the polypeptide fragment is HA2."],["<SOH> BACKGROUND INFORMATION <EOH>Cancer is the second leading cause of death in the United States, accounting for over one half million deaths per year.","(National Vital Statistics Report, 1998, Vol.","48, No.","11).","The total economic cost associated with cancer has been estimated to be over $100 billion dollars annually.","(Brown, M. L. et al.","In Cancer Epidemiology and Prevention, 1996).","There is currently no cure for the disease, but several lines of research appear promising.","Of these, research directed at preventing angiogenesis offers the most hope.","Angiogenesis involves a complex biochemical cascade of events that leads to new blood vessel formation in, for example, developing tumors.","For cells to survive, each must have some communication, direct or indirect with the existing vasculature in order to obtain nutrients and oxygen, and to offload metabolic waste products.","Active vascularization is normally observed following injury to a tissue, during development, or in response to ovulation in females.","However, abnormal rapid proliferation of blood vessels is also observed in areas where cancerous masses have developed.","Because the rate of cell mass growth is limited by the degree of vascularization present, cancer cells release chemical substances into the surrounding environment to induce nearby, nourishing blood vessels to grow toward the proliferating cancer cell mass.","For example, angiopoietin-1 basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF) and other substances are released from cancer cells to coax surrounding blood vessels to grow collateral vessels toward the tumor.","Once vascularized, a tumor mass may grow locally, or may metastasize and begin to spread through the bloodstream and lymphatic system to other parts of the body, causing significant damage.","For example, vascularization of the retina in diabetic retinopathy can lead to retinal detachment resulting in blindness.","Metastasis is a hallmark for malignancy, which in extreme cases may lead to rapid death of an individual.","Ironically, some tumors may also secrete angiostatic substances to inhibit tumor growth (Chen et al., Cancer Res.","1995, 55, 4230-4233; O'Reilly, Cell 1997, Jan. 24 (88):277-285).","Thus, it appears that in healthy individuals angiogenesis associated with tumor growth may be regulated by a fine balance between the release of angiogenic factors and the release of angiostatic factors.","It is believed that by blocking the process of angiogenesis, tumor growth can be suspended, which in turn would lead to cancer remission.","Most early research directed at preventing angiogenesis involved exposing various cell lines to angiostatic compounds and assessing the degree of proliferation either in vivo or in vitro.","The National Cancer Institute, for example, uses proliferation, migration and cord formation assays in HUVE cells for its anti-angiogenesis testing.","Several angiostatic agents that function to prevent the proliferation of cancer cells have been isolated and tested.","For example, administration of Angiostatin has been shown to suppress vascular endothelial cell proliferation, thereby reducing the size and lethality of tumors (Folman J., Forum Genova 1999 July-Dec. 9 (3 Suppl.","3): 5962).","Recombinant Endostatin (baculovirus) has been used to inhibit the proliferation of bovine capillary endothelial cells.","(O'Reilly, et al., Cell, 1994, Oct. 21; 79(2):185-8).","Until recently, angiostatic compounds have included only those substances capable of preventing proliferation of cells.","However, a growing body of evidence demonstrates that agents which inhibit proliferation via cellular detachment from tumor masses perform an analogous function.","Recent studies have been directed at interfering or disrupting the mechanisms involved in cell-cell or cell-matrix binding as a means to reduce or eliminate cancerous growth.","Cells will not proliferate if they are not first attached to a surface.","For example, impaired cell-matrix contact leads to anoikis (epithelial apoptosis) (Vitale et al., FEBS Lett.","1999 462 (1-2:57-60); Attwell et al., Oncogene 2000 19(33):3811-5; Rosen, J.","Cell.","Biol 2000 149(2):447-56); Rytomaa et al., Curr.","Biol.","1999 9(18):1043-6 or endothelial apoptosis (Erdreich-Epstein, Cancer Res.","2000 Feb. 1; 60(3):71221).","This anchorage dependence is mediated, in part, by cell surface molecules known as integrins.","See e.g., Erdreich-Epstein et al., Cancer Res., 2000 Feb. 1; 60(3):712-21; Lee & Juliano, Mol.","Biol.","Cell, 2000 Jun.","11(6):1973-87; Kawahara, J.","Cancer Res.","Clin.","Oncol, 1995 1212 (3):133-40; Lee, Mol.","Biol.","Cell 2000 11(6):1973-87; Ruoslahti, Kidney Int.","1997 51(5):1413-7; Brassard et al., Exp.","Cell.","Res.","1999 251 (1):33-45; Kottke et al., J. Biol.","Chem.","1999 274(22):15927-36.Anti-angiogenic approaches are the most recent and promising avenue in cancer treatment.","Agents capable of blocking vascularization of neoplastic tissue can prevent subsequent growth of transformed tissue and can lead to existing tissue remission.","Anti-angiogenic activity has been detected for several endogenous factors.","For example, combrestatin A-4 disodium phosphate (CA4DP) (Dark, Cancer Res.","1997 May 15; 57 (10):1829-34), a purified human PSA compound (WIPO PCT publication No.","WO 99/60984), Endostatin (add, et al., Biochem.","Biophys.","Res.","Commun.","1999 Sep. 24:263(2):3405), so (AIbini, Faseb J.","1999 13(6):647-55), Epidermal Growth Factors (EGF) (Kotke, et al., J. Bio.","Cam 1999 May 28;274(22):15927-36), and angiostatin (Stack, Biochem J., 1999 340 (pt1):77-84) have anti-angiogenic activity.","For many years, scientists have been in search of therapeutics that can be used to prevent periodontal diseases, including gum infections and tooth decay.","One organism that has been identified as a potential etiologic agent of such pathologies as gingivitis and periodontal disease is the pathogen Porphyromonas gingivalis .","Sequences from the pathogen have been cloned and sequenced.","Examples of such work can be found in U.S. Pat.","Nos.","5,824,791 and 5,830,710, both of which are hereby incorporated herein by reference in their entity.","Proteolytic enzymes of Porphyromonas gingivalis are the main tool for providing nutrients to these asaccharolytic bacteria and are also important virulence factors.","These enzymes have been shown to degrade basement membrane matrix proteins (Uitto, Oral Microbiology and Immunology 1988 3:97-102), (Smalley, Arch.","Oral Biol.","1988 33 (5):323-9) and purified P. gingivalis cysteine protease has been shown to disrupt the basement membrane of human carcinoma monolayer (Shah, J. Periodontol.","1992 63(1):44-51).","Targeted disruption of fibronectin-integrin interactions in human gingival fibroblasts has been demonstrated by the RI protease of P. gingivalis W50 (Scragg, et al., Infect.","Immun.","1999 67(4):1837-43).","Other studies have examined the function of P. gingivalis extract on cell-cell and cell-matrix bonds.","Cell detachment from each other and from the underlying surface correlates with the cysteine-dependent proteolytic activity of P. gingivalis (Johansson, Eur.","J.","Oral.","Sci.","1998 106(4):863-71).","B1-integrin, occluding and E-cadherin are targeted by P. gingivalis proteolytic activity in canine epithelial cells (Katz, et al., Infect.","Immun.","2000 68(3):1441-9)."],["<SOH> SUMMARY OF THE INVENTION <EOH>It is an object of the invention to provide compositions and methods for treatment, prevention or amelioration of angioproliferitive conditions.","This and other objects of the invention are provided by one or more of the embodiments described below.","One embodiment of the invention provides a composition for the t ent, prevention, or amelioration of an angioproliferitive condition.","The composition comprises a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide has anti-angiogenic activity, and a pharmaceutically acceptable excipient.","The polypeptide can be selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.","The polypeptide can have at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, prtT, or tla polypeptide and has anti-angiogenic activity.","The polypeptide can be a biologically functional homolog, or isoform of a rgpA, rgpB, kgp, hag, prtT or tla polypeptide.","The polypeptide can be a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cystine protease polypeptide, or Porphyromonas gingivalis hemagglutinin polypeptide, such as HA2.Such a fragment has anti-angiogenic activity.","An angioproliferative condition can be carcinoma, sarcoma, melanoma, benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthritis, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, arteriosclerosis, scleroderma, hypertrophic scars, cat scratch disease, Helicobacter pylori ulcers, capillary proliferation within atherosclerotic plaque, or a combination thereof.","Another embodiment of the invention provides a method for the treatment, prevention, or amelioration of an angioproliferative condition.","The method comprises administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a patient in need thereof, wherein the polypeptide has anti-angiogenic activity, whereby the angioproliferative condition is treated, prevented or ameliorated.","The method can comprise contacting a vasculature supplying a biological structure affected by the angioproliferative condition with the polypeptide.","The polypeptide can be contacted with a basolateral surface of the vasculature.","Yet another embodiment of the invention provides a method for potentiating effects of a chemotherapeutically effective agent.","The method comprises administering a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide has disrupts cell-cell adhesion, cell-matrix adhesion, or both; and a chemotherapeutically effective agent to a patient.","Effects of the chemotherapeutically effective agent arm potentiated.","Still another embodiment of the invention provides a method for preventing the formation of now vasculature required for implantation or sustenance of a fertilized mammalian ovum.","The method comprises administering a Porphyromonas gingivalis amine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a mammal, wherein the polypeptide has anti-angiogenic activity, whereby the formation of new vasculature required for implantation or sustenance of a fertilized mammalian ovum is prevented.","Even another embodiment of the invention provides a pharmaceutical composition for facilitating passage of compounds through a blood-brain barrier comprising a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, a Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, and a pharmaceutically acceptable excipient.","The pharmaceutical composition can further include a compound to be passed through the blood-brain barrier.","Another embodiment of the invention provides a method of delivering a compound through a blood-brain barrier of a patient comprising administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof and the compound, whereby the compound is passed through the blood-brain barrier.","Still another embodiment of the invention provides a method of degrading a tumor.","The method comprises administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a patient having a tumor, wherein the polypeptide degrades cell-cell bonds, cell-matrix bonds, or both cell-cell bonds and cell-matrix bonds of the tumor, whereby the tumor is degraded.","Therefore, the present invention discloses anti-angiogenic compositions of mater and methods of their use, which are capable of disrupting endothelial tissue growth and proliferation.","The methods involve local or systemic application of a P. gingivalis polypeptide to a targeted tissue.","The disclosed polypeptides can inhibit angiogenesis associated with malignant tumor proliferation by, for example, disrupting endothelial layer cell-cell and cell-matrix adhesion bonds.","Use of these compounds and methods have advantages over conventional treatments such as chemotherapy, because it targets only growing vessels, while leaving intact vessels unaffected.","The present invention provides a novel strategy that uses protein, peptide and nucleic acid sequences of Porphyromonas gingivalis to treat or prevent angioproliferative conditions."],["PRIORITY This application claims priority to U.S. Ser.","No.","09/849,115, filed on May 5, 2001, which is incorporated by reference in its entirety herein FIELD OF THE INVENTION The invention provides compositions and methods for the treatment, prevention and amelioration of angioproliferative conditions.","BACKGROUND INFORMATION Cancer is the second leading cause of death in the United States, accounting for over one half million deaths per year.","(National Vital Statistics Report, 1998, Vol.","48, No.","11).","The total economic cost associated with cancer has been estimated to be over $100 billion dollars annually.","(Brown, M. L. et al.","In Cancer Epidemiology and Prevention, 1996).","There is currently no cure for the disease, but several lines of research appear promising.","Of these, research directed at preventing angiogenesis offers the most hope.","Angiogenesis involves a complex biochemical cascade of events that leads to new blood vessel formation in, for example, developing tumors.","For cells to survive, each must have some communication, direct or indirect with the existing vasculature in order to obtain nutrients and oxygen, and to offload metabolic waste products.","Active vascularization is normally observed following injury to a tissue, during development, or in response to ovulation in females.","However, abnormal rapid proliferation of blood vessels is also observed in areas where cancerous masses have developed.","Because the rate of cell mass growth is limited by the degree of vascularization present, cancer cells release chemical substances into the surrounding environment to induce nearby, nourishing blood vessels to grow toward the proliferating cancer cell mass.","For example, angiopoietin-1 basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF) and other substances are released from cancer cells to coax surrounding blood vessels to grow collateral vessels toward the tumor.","Once vascularized, a tumor mass may grow locally, or may metastasize and begin to spread through the bloodstream and lymphatic system to other parts of the body, causing significant damage.","For example, vascularization of the retina in diabetic retinopathy can lead to retinal detachment resulting in blindness.","Metastasis is a hallmark for malignancy, which in extreme cases may lead to rapid death of an individual.","Ironically, some tumors may also secrete angiostatic substances to inhibit tumor growth (Chen et al., Cancer Res.","1995, 55, 4230-4233; O'Reilly, Cell 1997, Jan. 24 (88):277-285).","Thus, it appears that in healthy individuals angiogenesis associated with tumor growth may be regulated by a fine balance between the release of angiogenic factors and the release of angiostatic factors.","It is believed that by blocking the process of angiogenesis, tumor growth can be suspended, which in turn would lead to cancer remission.","Most early research directed at preventing angiogenesis involved exposing various cell lines to angiostatic compounds and assessing the degree of proliferation either in vivo or in vitro.","The National Cancer Institute, for example, uses proliferation, migration and cord formation assays in HUVE cells for its anti-angiogenesis testing.","Several angiostatic agents that function to prevent the proliferation of cancer cells have been isolated and tested.","For example, administration of Angiostatin has been shown to suppress vascular endothelial cell proliferation, thereby reducing the size and lethality of tumors (Folman J., Forum Genova 1999 July-Dec. 9 (3 Suppl.","3): 5962).","Recombinant Endostatin (baculovirus) has been used to inhibit the proliferation of bovine capillary endothelial cells.","(O'Reilly, et al., Cell, 1994, Oct. 21; 79(2):185-8).","Until recently, angiostatic compounds have included only those substances capable of preventing proliferation of cells.","However, a growing body of evidence demonstrates that agents which inhibit proliferation via cellular detachment from tumor masses perform an analogous function.","Recent studies have been directed at interfering or disrupting the mechanisms involved in cell-cell or cell-matrix binding as a means to reduce or eliminate cancerous growth.","Cells will not proliferate if they are not first attached to a surface.","For example, impaired cell-matrix contact leads to anoikis (epithelial apoptosis) (Vitale et al., FEBS Lett.","1999 462 (1-2:57-60); Attwell et al., Oncogene 2000 19(33):3811-5; Rosen, J.","Cell.","Biol 2000 149(2):447-56); Rytomaa et al., Curr.","Biol.","1999 9(18):1043-6 or endothelial apoptosis (Erdreich-Epstein, Cancer Res.","2000 Feb. 1; 60(3):71221).","This anchorage dependence is mediated, in part, by cell surface molecules known as integrins.","See e.g., Erdreich-Epstein et al., Cancer Res., 2000 Feb. 1; 60(3):712-21; Lee & Juliano, Mol.","Biol.","Cell, 2000 Jun.","11(6):1973-87; Kawahara, J.","Cancer Res.","Clin.","Oncol, 1995 1212 (3):133-40; Lee, Mol.","Biol.","Cell 2000 11(6):1973-87; Ruoslahti, Kidney Int.","1997 51(5):1413-7; Brassard et al., Exp.","Cell.","Res.","1999 251 (1):33-45; Kottke et al., J. Biol.","Chem.","1999 274(22):15927-36.Anti-angiogenic approaches are the most recent and promising avenue in cancer treatment.","Agents capable of blocking vascularization of neoplastic tissue can prevent subsequent growth of transformed tissue and can lead to existing tissue remission.","Anti-angiogenic activity has been detected for several endogenous factors.","For example, combrestatin A-4 disodium phosphate (CA4DP) (Dark, Cancer Res.","1997 May 15; 57 (10):1829-34), a purified human PSA compound (WIPO PCT publication No.","WO 99/60984), Endostatin (add, et al., Biochem.","Biophys.","Res.","Commun.","1999 Sep. 24:263(2):3405), so (AIbini, Faseb J.","1999 13(6):647-55), Epidermal Growth Factors (EGF) (Kotke, et al., J. Bio.","Cam 1999 May 28;274(22):15927-36), and angiostatin (Stack, Biochem J., 1999 340 (pt1):77-84) have anti-angiogenic activity.","For many years, scientists have been in search of therapeutics that can be used to prevent periodontal diseases, including gum infections and tooth decay.","One organism that has been identified as a potential etiologic agent of such pathologies as gingivitis and periodontal disease is the pathogen Porphyromonas gingivalis.","Sequences from the pathogen have been cloned and sequenced.","Examples of such work can be found in U.S. Pat.","Nos.","5,824,791 and 5,830,710, both of which are hereby incorporated herein by reference in their entity.","Proteolytic enzymes of Porphyromonas gingivalis are the main tool for providing nutrients to these asaccharolytic bacteria and are also important virulence factors.","These enzymes have been shown to degrade basement membrane matrix proteins (Uitto, Oral Microbiology and Immunology 1988 3:97-102), (Smalley, Arch.","Oral Biol.","1988 33 (5):323-9) and purified P. gingivalis cysteine protease has been shown to disrupt the basement membrane of human carcinoma monolayer (Shah, J. Periodontol.","1992 63(1):44-51).","Targeted disruption of fibronectin-integrin interactions in human gingival fibroblasts has been demonstrated by the RI protease of P. gingivalis W50 (Scragg, et al., Infect.","Immun.","1999 67(4):1837-43).","Other studies have examined the function of P. gingivalis extract on cell-cell and cell-matrix bonds.","Cell detachment from each other and from the underlying surface correlates with the cysteine-dependent proteolytic activity of P. gingivalis (Johansson, Eur.","J.","Oral.","Sci.","1998 106(4):863-71).","B1-integrin, occluding and E-cadherin are targeted by P. gingivalis proteolytic activity in canine epithelial cells (Katz, et al., Infect.","Immun.","2000 68(3):1441-9).","SUMMARY OF THE INVENTION It is an object of the invention to provide compositions and methods for treatment, prevention or amelioration of angioproliferitive conditions.","This and other objects of the invention are provided by one or more of the embodiments described below.","One embodiment of the invention provides a composition for the t ent, prevention, or amelioration of an angioproliferitive condition.","The composition comprises a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide has anti-angiogenic activity, and a pharmaceutically acceptable excipient.","The polypeptide can be selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.","The polypeptide can have at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, prtT, or tla polypeptide and has anti-angiogenic activity.","The polypeptide can be a biologically functional homolog, or isoform of a rgpA, rgpB, kgp, hag, prtT or tla polypeptide.","The polypeptide can be a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cystine protease polypeptide, or Porphyromonas gingivalis hemagglutinin polypeptide, such as HA2.Such a fragment has anti-angiogenic activity.","An angioproliferative condition can be carcinoma, sarcoma, melanoma, benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthritis, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, arteriosclerosis, scleroderma, hypertrophic scars, cat scratch disease, Helicobacter pylori ulcers, capillary proliferation within atherosclerotic plaque, or a combination thereof.","Another embodiment of the invention provides a method for the treatment, prevention, or amelioration of an angioproliferative condition.","The method comprises administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a patient in need thereof, wherein the polypeptide has anti-angiogenic activity, whereby the angioproliferative condition is treated, prevented or ameliorated.","The method can comprise contacting a vasculature supplying a biological structure affected by the angioproliferative condition with the polypeptide.","The polypeptide can be contacted with a basolateral surface of the vasculature.","Yet another embodiment of the invention provides a method for potentiating effects of a chemotherapeutically effective agent.","The method comprises administering a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide has disrupts cell-cell adhesion, cell-matrix adhesion, or both; and a chemotherapeutically effective agent to a patient.","Effects of the chemotherapeutically effective agent arm potentiated.","Still another embodiment of the invention provides a method for preventing the formation of now vasculature required for implantation or sustenance of a fertilized mammalian ovum.","The method comprises administering a Porphyromonas gingivalis amine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a mammal, wherein the polypeptide has anti-angiogenic activity, whereby the formation of new vasculature required for implantation or sustenance of a fertilized mammalian ovum is prevented.","Even another embodiment of the invention provides a pharmaceutical composition for facilitating passage of compounds through a blood-brain barrier comprising a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, a Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, and a pharmaceutically acceptable excipient.","The pharmaceutical composition can further include a compound to be passed through the blood-brain barrier.","Another embodiment of the invention provides a method of delivering a compound through a blood-brain barrier of a patient comprising administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof and the compound, whereby the compound is passed through the blood-brain barrier.","Still another embodiment of the invention provides a method of degrading a tumor.","The method comprises administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a patient having a tumor, wherein the polypeptide degrades cell-cell bonds, cell-matrix bonds, or both cell-cell bonds and cell-matrix bonds of the tumor, whereby the tumor is degraded.","Therefore, the present invention discloses anti-angiogenic compositions of mater and methods of their use, which are capable of disrupting endothelial tissue growth and proliferation.","The methods involve local or systemic application of a P. gingivalis polypeptide to a targeted tissue.","The disclosed polypeptides can inhibit angiogenesis associated with malignant tumor proliferation by, for example, disrupting endothelial layer cell-cell and cell-matrix adhesion bonds.","Use of these compounds and methods have advantages over conventional treatments such as chemotherapy, because it targets only growing vessels, while leaving intact vessels unaffected.","The present invention provides a novel strategy that uses protein, peptide and nucleic acid sequences of Porphyromonas gingivalis to treat or prevent angioproliferative conditions.","BRIEF DESCRIPTION OF THE FIGURES FIG.","1 demonstrates percent detachment of active Human Umbilical Vein Endothelial Cells (HUVEC) after 24 hours of treatment with a protein extract from P. gingivalis.","Represented are mean values from triplicate experiments.","FIG.","2 demonstrates percent detachment of active HUVEC after 48 hours of treatment with a protein extract from P. gingivalis.","FIG.","3 demonstrates percent detachment of quiescent HUVEC after 24 hours of treatment with a protein extract from P. gingivalis.","FIG.","4 demonstrates percent detachment of quiescent HUVEC after 48 hours of treatment with a protein extract from P. gingivalis.","FIG.","5 demonstrates percent detachment of active human non-small cell lung carcinoma cell line (A549) after 24 hous of treatment with a protein extract from P. gingivalis.","FIG.","6 demonstrates percent detachment of active human non-small cell lung carcinoma cell line (A549) after 48 hours of treatment with protein extract from P. gingivalis.","FIG.","7 demonstrates percent detachment of quiescent human non-small cell lung carcinoma cell line (A549) after 24 hours of treatment with a protein extract from P. gingivalis.","FIG.","8 demonstrates percent detachment of quiescent human non-small cell lung carcinoma cell line (A549) after 48 hours of treatment with a protein extract from P. gingivalis.","FIG.","9 demonstrates the reduction in human vascular endothelial cell migration after exposure to a protein extract from P. gingivalis over a 24 hour period.","FIG.","10 demonstrates detachment of human non-small cell lung carcinoma cell line (A549) after treatment with a proteinase extract of P. gingivalis alone and with an extract of Bacteroides fragilis expressing PrtP protease from P. gingivalis.","FIG.","11 is a table showing the degree of proliferation inhibition of a HUH7 cell line exposed to P. gingivalis and E. coli extract, and to P. gingivalis cells in the presence or absence of inhibitors.","FIG.","12A-E shows the results of reactivity of a HUH7 cell line with anti-occludin antibodies.","FIG.","12A depicts non-treated control HUH7 cells.","FIG.","12B depicts the HUH7 cells treated with P. gingivalis extract.","FIG.","12C depicts HUH7 cells treated with P. gingivalis extract in the presence of the inhibitor TLCK.","FIG.","12D depicts results of treatment with heat-treated P. gingivalis extract.","FIG.","12E depicts HUH7 cells treated with a control E. coli extract; the occludin network is intact.","FIG.","13A-D show the result of reactivity of a HUH7 cell tine with an anti-pan cadherin antibody.","FIG.","13A depicts non-treated control HUH7 cells.","FIG.","13B depicts HUH7 cells treated with P. gingivalis extract.","FIG.","13C depicts HUH7 cells treated with P. gingivalis extract in the presence of inhibitor TLCK.","FIG.","13D depicts HUH7 cells treated with heat inactivated P. gingivalis extract.","FIG.","14A-C shows proliferation inhibition of a HUVEC polarized cell line.","FIG.","14A depicts non-treated, control polarized human endothelial cell layer, ECV-304.FIG.","14B shows ECV-304 cells treated basolaterally (lower chamber) with a 60% fraction of P. gingivalis culture liquid proteins.","FIG.","14C depicts the results of ECV-304 cells lumenally treated with a 60% fraction of P. gingivalis culture liquid proteins.","FIG.","15 demonstrates total cell number reduction as result of treatment with P. gingivalis strain W83 extract.","FIG.","16 demonstrates that detachment of HUVE cells is reduced by inhibitors or heating of SPF.","FIG.","17 demonstrates transendothelial resistance of HUVE cells upon treatment with P. gingivalis soluble protein fraction (SPF) over a period of 6 hours.","FIG.","18 demonstrates transendothelial resistance of HUVE cells upon treatment with P. gingivalis soluble protein fraction (SPF) over a period of 8 days.","FIG.","19A-C shows contrast micrographs of HUVE cell layers that were treated with P. gingivalis SPF or mock-treated at day 8.FIG.","19A shows treated cells.","FIG.","19B shows BL-treated cells.","FIG.","19C shows mock-treated control cells.","FIG.","20A-C show contrast micrographs of HUVE cells treated with extracts of E. coli.","FIG.","20A shows HUVE cells treated with control E. coli host extract.","FIG.","20B shows control mock-treated cells.","FIG.","20C shows cells treated with extracts of E. coli host cells that express a 15 kDa internal fragment of a P. gingivalis HagA repeat.","FIG.","21 shows proliferation inhibition of HUVE cells treated with HA2 15-kDa P. gingivalis protein cloned in E. coli.","(E. coli 0.5: control E. coli protein extract at 0.5 mg/ml.","E. coli HA2 0.5: E. coli protein extract containing HA2, the 15-kDa polypeptide at 0.5 mg/ml.","FIG.","22 shows potential pathways for leakage from blood vessels in tumors.","Intercellular openings (arrows) between lining cells of a murine tumor vessel viewed by scanning EM.","FIGS.","22A and B: Multiple large intercellular openings (arrow, FIG.","22A) and three smaller transcellular holes (arrows, FIG.","22B) in branched lining cells of a tumor vessel.","The intercellular openings are much larger than the holes.","The boxed region in FIG.","22A is shown at higher magnification in FIG.","22B.","DETAILED DESCRIPTION OF THE INVENTION “Angiogenesis” or “angioproliferation” means conditions of rapid development of vascular supply to a particular organ or biological site.","The rapid development can be uncontrolled and/or pathogenic, as in the development of a tumor.","However, angiogenisis or angioproliferation can also be associated with non-pathological conditions such as angiogenesis which occurs upon implantation of a fertilized ova.","An anti-angiogenic or anti-angioproliferative composition is capable of reducing or preventing angiogenisis or angioproliferation.","The present invention is directed to arginine- or lysine-specific cysteine protease polypeptides and hemagglutinin polypeptides derived from P. gingivalis.","The polypeptides can be used to treat or prevent angioproliferative conditions, including but not limited to melanoma, sarcoma, and carcinomas including, for example, breast, colon, lung and prostate carcinomas.","Additional pathologies susceptible to treatment according to the present invention include benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthritis, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, scleroderma, hypertrophic scars, cat scratch disease, and Helicobacter pylori ulcer and capillary proliferation within atherosclerotic plaque.","Post-surgery adhesions are common complication of gynecologic and abdominal surgery.","Such complications can lead to infertility, ectopic pregnancy, chronic pain, prolonged recovery and intestinal obstruction.","Adhesions following surgery can be prevented by applying polypeptides of the invention at the end of the operation.","Furthermore, the compositions of the invention can be used to control non-pathogenic angiogenic conditions, such as using the compositions as a contraceptive to prevent implantation of fertilized ova.","Polypeptides of the Invention Compositions of the invention comprise a substantially purified Porphymonas gingivalis arginine-specific cysteine protease polypeptide, a Porphymonas gingivalis lysine-specific cysteine protease polypeptide, a Porphymonas gingivalis hemagglutinin polypeptide or combinations thereof.","These polypeptides are expressed from members of the P. gingivalis protease and hemagglutinin gene family.","See e.g., Curtis et al., J. Periodont.","Res.","34:494 (1999).","“Substantially purified” refers to polypeptides that are removed from their natural environment, isolated or separated, and are at least 60% free, preferably 75% free, and most preferably 90% free from other components with which they are naturally associated.","Polypeptides of the invention include rgpA arginine-specific cysteine proteases.","P. gingivalis rgpA proteases are a group of proteases produced by genes of a homologous loci in different of P. gingivalis.","Examples of the rgpA proteases are: prpR1 (Accession number X82680), rgp-1 (Accession number U15282), rgpA (Accession number D26470), prtR (Accession number L26341), prtH (Accession number L27483), and hagE (Accession number AF026946), which is similar to rgp (Accession number A55426).","Isoforms of rgpA proteases are also part of the invention and include, for example, RgpA(cat), mt-RgpA(cat) and HRgpA isoforms.","See, e.g., Curtis et al., J. Periodont.","Res.","34:494 (1999).","P. gingivalis rgpB arginine-specific proteases are also polypeptides of the invention Examples of rgpB proteases include prtRII (Accession number AF007124), rgp-2 (Accession number U85038), rgpB (Accession number D64081), and prR2 (Rangarajan, Mol Microbiol 1997 Mar.","23(5):955-65).","The rgpB proteases are all highly related and are derived from homologous genes which occur in different strains of P. gingivalis.","Isoforms of rgpB proteases are also part of the invention and include, for example, RgpB and mt-RgpB isoforms.","See e.g., Curtis et al., J. Periodont.","Res.","34:494 (1999).","Lysine-specific proteases of P. gingivalis include kgp proteases.","kgp proteases include, for example, prtK (Accession number U75366), kgp (Accession numbers U54691; D83258), prtP (Accession numbers U42210; AF017059), kgp(381)-hagD (Accession number U68468).","The kgp proteases are all highly related and are derived from homologous genes in different stains of P. gingivalis.","Isoforms of kgp proteases are also included in the invention.","Hemagglutinin polypeptides of the invention include hagA, hagB, hagC, hagD, hagE, tla (e.g., Accession number Y07618) and prtT (Accession number S75942; M83096).","See also, e.g., U.S. Pat.","Nos.","5,824,791 and 5,830,710 and Aduse-Opoku et al., J. Bacteriol.","179:4778-4788 (1997).","As disclosed in U.S. Pat.","No.","5,824,791, a hag gene can be a 7887 bp molecule, which encodes a gene product of 2628 amino acids.","Within the gene product, there are four repeat segments: HArep1, HArep2, HArep3 and HArep4.Bach of these segments contains a 15-kDa fragment, HA2, that possesses anti-angiogenic activity and is found in the secreted protein fraction.","HA2 is instrumental in production of necrosis, which leads to periodontal disease.","HA2 is located within a hagA sequence between amino acids 683 and 819 in the first repeat and is similarly located in the other repeats (see Han et al., Infect.","Immun.","Page 4002, FIG.","2, U.S. Pat.","Nos.","5,824,791 and 5,830,710).","P. gingivalis hemagglutinin polypeptides have cysteine protease activity against arginine containing substrates.","Nishikata and Yoshimura, Biochem Bio.","Phys.","Res.","Comm 178:336-40 (1991).","P. gingivalis hemagglutinins also function as attachment factors and the substrate binding site is responsible for attachment to erythrocytes.","Hemagglutinins also contain “peptidase C25 family” activity.","The entire hagA, hagB, hagC, hagD, hagE, prtT or tla polypeptide can be used to produce anti-angiogenic effects.","Alternatively, any of the HArep sequences or HA2 can be used Additionally, compounds with similar activity can also be developed which have equal or greater potency than polypeptides of the invention, and such compounds come within the scope of this invention.","For example, one of skill in the art can design peptidomimetics using, for example, directed protein evolution or by minimizing the size of the polypeptide so that improved or smaller versions of the polypeptide are developed.","Polypeptides of the invention include polypeptides produced from any P. gingivalis rgpA, rgpB, kgp, hag, prtT or tla gene.","It is well known in the art that the proteases and hemagglutinins of P. gingivalis are produced from a family of protease and hemagglutinin genes that are highly related.","This gene family is known to vary from strain to strain of P. gingivalis.","See e.g., Curtis et al., J. Periodont.","Res.","34:494 (1999).","The invention includes all polypeptides produced from the rgpA, rgpB, kgp, hag, prtT or tla gene family that have anti-angiogenic activity.","One of skill in the art could determine whether a protease or hemagglutinin gene or polypeptide fell within the rgpA, rgpB, kgp, hag, prtT or tla family by comparing the nucleotide or protein sequence of the gene or protein in question to the known sequences of these genes and proteins.","Curtis et al.","(J. Peridont.","Res.","34:464 (1999)) describes these genes families and discloses examples of genes that fall within the P. gingivalis protease and hemagglutinin gene family.","U.S. Pat.","Nos.","5,824,791 and 5,830,710 further describes genes that fall within the hag family.","All of these related polypeptides have cysteine protease activity and sequence homology and as such, can be classified into the protease-hemagglutinin family.","Polypeptides of the invention can either be full-length polypeptides or fragments of polypeptides.","For example, fragments of polypeptides of the invention can comprise about 10, 25, 50, 100, 200, 250, 500, 750, or 1,000 amino acids of polypeptides of the invention Percent sequence identity ha an art recognized meaning and there are a no of methods to measure identity between two polypeptide or polynucleotide sequences.","See, e.g., Lesk, Ed, Computational Molecule Biology, Oxford University Press, New York, (1988); Smith, Ed, Biocomputing: Informatics And Genome Projects, Academic Press, New York, (1993); Griffin & Griffin, Eds., Computer Analysis Of Sequence Data, Part I, Humana Press, New Jersey, (1994); von Heinje, Sequence Analysis In Molecular Biology, Academic Press, (1987); and Gribskov & Devereux, Eds., Sequence Analysis Primer, M Stockton Press, New York, (1991).","Methods for aligning polynucleotides or polypeptides are codified in computer programs, including the GCG program package (Devereux et al., Nuc.","Acids Res.","12:387 (1984)), BLASTP, BLASTN, FASTA (Atschul et al., J. Mole Biol.","215:403 (1990)), and Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wis. 53711) which uses the local homology algorithm of Smith and Waterman (Adv.","App.","Math., 2:482-489 (1981)).","For example, the computer program ALIGN which employs the FASTA algorithm can be used, with an affine gap search with a gap open penalty of −12 and a gap extension penalty of −2.When using any of the sequence alignment programs to determine whether a particular sequence is, for instance, about 95% identical to a reference sequence, the parameters are set such that the percentage of identity is calculated over the full length of the reference polynucleotide and that gaps in identity of up to 5% of the total number of nucleotides in the reference polynucleotide are allowed.","Amino acid sequences having at least about 75, preferably at least about 90, 95, 96, 98, or 99% sequence identity to amino acid sequences of P. gingivalis rgpA, rgpB, kgp, hag, prtT or tla polypeptides and have anti-angiogenic activity are also polypeptides of the invention.","Also included in the invention are polypeptides having amino acid sequences which are at least about 75, preferably at least about 90, 95, 96, 98, or 99% sequence identity toamino acid sequences of P. gingivalis rgpA, rgpB, kgp, hag, prtT or tla proteins that are produced by an organism other than P. gingivalis and have anti-angiogenic activity.","Polypeptides of the invention also are biologically functional homologs, analogs and isoforms of rgpA, rgpB, kgp, hag, prtT or tla polypeptides.","A biologically functional homolog is a polypeptide that has sequence identity to an analogous polypeptide from another species.","Homologs can be naturally occurring or can be a polypeptide that does not occur in nature.","An isoform is encoded by a distinct mRNA splice variant and can be a naturally occurring polypeptide or can be a polypeptide that does not occur in nature.","An analog is a polypeptide having alterations involving one or more amino acid insertions and deletions and/or conservative amino acid substitutions.","A homolog, analog or isoform of the invention comprises at least about 75, 90, 95, 96, 98 or 99% sequence identity to a rgpA, rgpB, kgp, hag, prtT or tla polypeptide.","A biologically functional homolog, analog or isoform has at least one biological activity of a rgpA, rgpB, kgp, hag, prtT or tla polypeptide.","That is, they are capable of preventing endothelial cell proliferation, disruption of vascular endothelium, promoting cellular detachment, inhibiting migration of endothelial cells, blocking formation of new blood vessels, or destroying existing blood vessels feeding, for example, tumors.","A biologically functional homolog or analog, isoform, or fragment can be tested for biological activity using, for example a cell proliferation/cell detachment assay (see Example 1), a migration inhibition assay (see Example 2), an occludin-stain junction assay (see Example 7), catherin-stain junction assay (see Example 8) proliferation inhibition of polarized cell assay (see Example 10), degradation of α5β1 integrin assay (see Example 11), or transendothelial resistance assay (see Example 12).","A biologically functional homolog, analog, isoform or fragment has about 85%, 90%, 95%, 98%, 99%, 100%, 105%, or 110% biological activity of a specific specie in question.","In one embodiment of the invention, a polypeptide of the invention is produced recombinantly.","A polynucleotide encoding a polypeptide of the invention can be introduced into a recombinant expression vector, which can be expressed in a suitable expression host cell system using techniques well known in the art.","A variety of bacterial, yeast, plant, mammalian, and insect expression systems are available in the art and any such expression system can be used.","Optionally, a polynucleotide encoding a polypeptide can be translated in a cell-free translation system.","A polypeptide of the invention can also be chemically synthesized.","If desired, a polypeptide can be produced as a fusion protein, which can also contain other amino acid sequences, such as amino acid linkers or signal sequences, as well as ligands useful in protein purification, such as glutathione-S-transferase, histidine tag, and staphylococcal protein A.","More than one polypeptide of the invention can be present in a fusion protein.","Biological Activity of Polypeptides of the Invention Polypeptides of the invention have anti-angiogenic biological activity.","That is, they are capable of preventing endothelial cell proliferation, disruption of vascular endothelium, promoting cellular detachment, inhibiting migration of endothelial cells, blocking formation of new blood vessels, or destroying existing blood vessels feeding, for example, tumors or combinations thereof.","Therefore, compositions of the invention are capable of inhibiting of tumor enlargement, decreasing tumor mass, and inhabiting, reducing, or preventing related biological processes associated with angiogenesis and vascular supply to a particular biological organ or location, or combinations thereof.","In addition, the polypeptides of the invention can be capable of disintegrating cell-to-matrix and cell-to-cell bonds in, for example, a tumor such that the tumor is reduced or eliminated.","A polypeptide of the invention has at least one of the above-mentioned biological activities.","Polypeptides of the invention can act on cell surface adhesion molecules (CAM's), from the basolateral side of the endothelium.","Polypeptides of the invention can inhibit angiogenesis by targeting integrin in cell-cell and cell-matrix adhesion bonds.","Of particular importance is the fact that vasculature supplying tumor tissues is aberrantly leaky.","Regular blood vessels are well formed and have a well-developed adhesion system to keep them together, while blood vessels supplying tumors are poorly formed and extremely leaky (Herlyn, Immunother.","1999 May, 22(3):185); see also, Hashizume, et al., Am.","J. Pathol.","2000 April 156(4):1363-80).","As a result, locally or systemically administered polypeptides of the invention leak out of the vasculature that supplies tumor tissue.","Access to the basolateral surface of the vasculature is thereby achieved, which results in disruption of the vascular at that location such that di-ation of the tumor occurs.","Therefore, compositions of the invention can disrupt angiogenesis without affecting integrity of normal blood vessels.","Methods of Treatment Compositions of the invention can be used to treat several conditions in mammals, including humans.","For example, angioproliferative conditions can be treated, prevented or ameliorated according the invention.","Methods of treatment comprise administering to a patient having an angioproliferative condition a pharmaceutically effective mount of a composition of the invention such that an anti-angiogenic effect is achieved Angioproliferative conditions include, for example, carcinoma, sarcoma, melanoma, benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthritis, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, atherosclerosis, scleroderma, hypertrophic scars, cat scratch disease, Helicobacter pylori ulcers, capillary proliferation within atherosclerotic plaque, or a combination thereof.","An angioproliferative condition can be selectively treated according to the invention by contacting vasculature supplying a biological structure affected by an angioproliferative condition with an anti-angiogenically effective amount of a composition of the invention.","Due to the known leakiness of vasculature supplying, for example, a tumor, local or systemic administration of a composition of this invention facilitates contact with the basolateral surface of said vasculature, including the endothelium, with no effect on normal tissues.","Compositions of the invention can also be used to potentiate the effects of a chemotherapeutically effective agent.","Such methods comprise co-administering a chemotherapeutically effective agent in the presence of a polypeptide of the invention effective to disrupt cell-cell adhesion, cell-matrix adhesion, or both.","Such co-administration can be in the form of a covalent complex, an ionic complex, a mixture simultaneous but separate administration, or administration within a relatively close temp sequence.","Appropriate chemotherapeutic agents include, but are not limited to doxorubicin, daunorubicin, doxorubicin, idarubicin, vincristine, 6-mercaptopurine, 6-thioguanine, methotrexate, cytoxan, cytarabine, L-asparaginase, busulfan, cyclophoshamide, melphalan, carmustine, lomustine, 5-fluorouracil methotrexate, fludarabine, bleomycin, docetaxel etoposide, vinorelbine, antibodies, and the like.","Polypeptides of the invention can also be used in contraception methods.","During pregnancy, the endometrial layer of the uterus becomes thickened and engorged with blood vessels upon implantation of a fertilized ovum.","Without a well-developed vasculature the fertilized ovum will not be sustained, and the endometrial layer will be sloughed-off in the form of menses, i.e., menstruation.","In one embodiment of the present invention, therapeutic compositions for use as contraceptives are provided.","In order to induce contraception, the internal vasulature of the uterus is contacted with a contraceptively effective amount of a polypeptide of the invention.","The mode of achieving bioavailability of polypeptides of the invention in this and other angioproliferative conditions can be, for example, through systemic or localized administration, such as intrauterine infusion.","Polypeptides of the invention can also be used to facilitate passage of compounds, such as pharmaceuticals, through the blood-brain barer of a mammal.","A polypeptide of the invention can be administered along with a compound to permeablize a blood-brain barrier and allow delivery of the compound thought the blood-brain barrier.","Such methods comprise co-administering a compound in the presence of a polypeptide of the invention effective to disrupt cell-cell adhesion, cell-matrix adhesion, or both.","Such co-administration can be in the form of a covalent complex, an ionic complex, a mixture, simultaneous but separate administration, or administration within a relatively close temporal sequence.","Pharmaceutical Compositions and Administration Thereof A pharmaceutically effective amount refers to an amount effective in treating an angiogenic condition in a mammalian patient, such as a human.","A pharmaceutically acceptable excipient is a non-toxic carrier, vehicle or adjuvant that can be administered to a patient, together with a polypeptide of this invention, and which does not destroy the pharmacological activity of the polypeptide.","Excipients, including carriers, vehicles and adjuvants are well known in the art See, e.g., Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Co., Easton Pa., 1985.A pharmaceutically effective amount is an amount of a polypeptide that achieves a specified functional result.","A dosage of a polypeptide of the invention can be determined using routine experimentation Pharmaceutical compositions of the invention can be administered by any number of routes including, but not limited to oral controlled release, intravenous, intramuscular, intra-arterial, intramedullary, intradermal intrathecal intraventricular, transdermal, subcutaneous, intraperitoneal, intranasal parenteral, topical, or rectal means.","Pharmaceutical compositions for oral administration can be formulated using pharmaceutically acceptable carriers well known in the art in dosages suitable for oral administration.","Such carriers enable the pharmaceutical compositions to be formulated as tablets, pills, dragees, capsules, liquids, gels, sips, slurries, suspensions, and the like, for ingestion by the patient.","Pharmaceutical formulations suitable for parenteral administration can be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks' solution, Ringer's solution, or physiologically buffered saline.","Aqueous injection suspensions can contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol or dextran.","Additionally, suspensions of the active compounds can be prepared as appropriate oily injection suspensions.","Suitable lipophilic solvents or vehicles include fatty oils such as sesame oily or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes.","Non-lipid polycationic amino polymers also can be used for delivery.","Optionally, the Z ion also can contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.","For topical or nasal administration, penetrants appropriate to the particular barrier to be permeated are used in the formulation.","Such penetrants are generally known in the art.","A polypeptide of the invention can also be delivered to a patient by administration of a polynucleotide encoding a polypeptide of the invention.","Preferably, injection of a P. gingivalis protease or hemagglutinin polynucleotide, which encodes a polypeptide of the invention, is used to treat, ameliorate, or prevent an angioproliferative condition.","In addition to the practical advantages of simplicity of construction and modification, injection of a polynucleotide results in the synthesis of a polypeptide of the invention in the host.","The polynucleotide is preferably delivered as “naked DNA” or in a vector.","A vector can be a plasmid, such as pBR322, pUC, or ColE1, or an adenovirus vector, such as an adenovirus Type 2 vector or Type 5 vector.","Optionally, other vectors can be used, including but not limited to Sindbis virus, simian virus 40, alphavirus vectors, poxvirus vectors, and cytomegalovirus and retroviral vectors, such as murine sarcoma virus, mouse mammary tumor virus, Moloney murine leukemia virus, and Rous sarcoma.","Minichromosomes such as MC and MC1, bacteriophages, phagemids, yeast artificial chromosomes, bacterial artificial chromosomes, virus particles, virus-like particles, cosmids (plasmids into which phage lambda cos sites have been inserted) and replicons (genetic elements that are capable of replication under their own control in a cell).","A polynucleotide can be, for example, injected intramuscularly to a mammal at a dose of 0.5, 0.75, 1.0, 1.5, 2.0, 2.5, 5 or 10 mg/kg.","In vivo delivery of polynucleotides (e.g., plasmid DNA) into results in the cellular uptake and expression of the polynucleotide into a desired polypeptide (Wolff, J.","A. et al., Science 247:1465-1468 (1990); Wheeler, C. J. et al, Proc.","Natl.","Acad.","Sci.","USA 93:11454-11459 (1996)).","The efficiency of in vivo polynucleotide administration can be increased using, for example, chemical agents or physical manipulations.","Such chemical agents include cellular toxins such as bupivacaine, cardiotoxin or barium chloride, polymers such as polyvinyl pyrolidone, polyvinyl alcohol polyethyleneimine, polyamidomine, and polyethylene glycol-polyethyleneimine-transferrin complexes that coat the DNA and protect it from DNases and enhance plasmid DNA-based expression or immune responses, particles that interact with the DNA and act as carriers and enhance DNA expression such as nanospheres, microspheres, dendrimers, collagen and polylactide co-glycolides, bulking agents such as sucrose, detergents such as sodium glycocholate, sodium deoxycholate, and beta-cyclodextrin, cationic or non-cationic lipids, DNA binding agents, or agents that enhance plasmid DNA transcription such as histone deacetylase inhibitor FR901228 or S-Bromo-cyclic AMP.","Physical manipulations include removal of nerves that control muscle contraction, electroporation, use of intravascular pressure, use of sutures coated with plasmid, use of sponges soaked with DNA as intramuscular depots to prolong DNA delivery, use of special needle-based injection methods, and of needleless-injectors tat propel the DNA into cells.","The pharmaceutical compositions of the present invention can be manufactured in a manner that is known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes.","The pharmaceutical composition can be provided as a salt and can be formed with many acids, including but not limited to, hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc.","Salts tend to be more soluble in aqueous or other protonic solvents than are the corresponding free base forms.","In other cases, the preferred preparation can be a lyophilized powder which can contain any or all of the following: 1-50 mM histidine, 0.1%-2% sucrose, and 2-7% mannitol at a pH range of 4.5 to 5.5, that is combined with buffer prior to use.","Further details on techniques for formulation and administration can be found in the latest edition of REMINGTON'S PHARMACEUTICAL SCIENCES (Maack Publishing Co., Easton, Pa.).","After pharmaceutical compositions have been prepared, they can be placed in an appropriate container and labeled for treatment of an indicated condition.","Such labeling would include amount, frequency, and method of administration.","Because of the specificity of polypeptides of the invention for rapidly developing vasculature, it is possible to define doses of the polypeptides that are non-toxic to the remainder of a patient, but which nonetheless provide a localized effect to achieve beneficial anti-angiogenic effects.","Dosages of a polypeptide of the invention can be about 0.01, 0.1, 1, 2, 5, or 10 mg/kg, of body weight, but one of ordinary skill in the art would recognize that for particular applications, it can be necessary to use either greater or lesser dosages.","If a polypeptide of the invention is found to generate an immune response in a patient, processes are known in the art for mimicking biological activity of proteins through development of minienzymes, DNA agents, or other small molecules, repeat administration of active site mimics including for chronic administration.","The above disclosure generally describes the present invention, and all patents, patent applications, scientific references cited in this disclosure are expressly incorporated herein.","A more complete understanding can be obtained by reference to the following specific examples which are provided for purposes of illusion only and are not intended to limit the scope of the invention.","EXAMPLE 1 Methods for Testing Human Endothelial or Carcinoma Cell Detachment Exposed to P. gingivalis Extract To determine the efficacy of P. gingivalis protease extracts in inhibiting cell proliferation, two cell lines were tested.","Proliferation inhibition was assessed by determining the detachment of tissue culture cells from their substrate.","A549 human non-small cell lung carcinoma cell line (maintained in RPMI-1640 medium (Gibco #11875-135) supplemented with L-Glu and Pen/Strep) and human endothelial cell line (HUVEC, ATCC #ECV-304), which was maintained in M199 medium, supplemented with L-Glutamine (0.05%, Gibco 25030-081), 0.1% Penicillin-Streptomycin (Gibco 15140-122) and 10% PBS (HyClone #SH30071.02, added after heat treatment at 56° C. for 30 min), were used as targets in a cell detachment assay.","Exponentially growing (Ex) and quiescent (Plateau) phases of cell culture growth were tested.","To obtain extract for activity tests, exponentially grown broth cultures of P. gingivalis strain W83 were pelleted, resuspended in 50 mM HEPES buffer (pH 7.5), and sonicated on ice for 2 min.","Following centrifugation at 14,000 rpm at 4 C, the supernatant was filtered through 0.22 μm non-protein-binding filter (Gelman) and stored at −80° C. until needed.","Protein concentration was measured using Sigma bicinchoninic acid reagent (#B-9643), BSA standard solution and spectrophotometer Shimadzu UV-1201.Six plates per sample with three plates added for non-treated control were used.","104 cells per 60-mm tissue culture plate were used to seed the sample.","Detached cells were counted by collecting the medium after treatment (24 or 48 hrs), washing the attached cells with calcium-free phosphate-buffered saline (PBS) and adding the wash to the medium.","The pellet was then resuspended in medium to obtain a countable number on the hemocytometer.","The 25-square (0.1 μl volume) count ×104 gives the cell count per ml.","Two readings were made of each of the three plates.","This number represented the detached cell count.","The attached cells were trypsinized and collected as above; the plate was washed with PBS and added to the tube.","After pelleting and resuspending, two readings were made per each of the three plates to obtain the attached cell count.","The mean value of the counts was taken, total number of cells (attached+detached) was obtained and the percentage of detached cells was calculated.","Different concentrations of P. gingivalis protein extract were used in the beginning to establish effective concentration to be used throughout.","The working concentration was chosen to be 0.4 mg total protein tact per ml cute medium.","FIG.","1 and FIG.","2 show non-confluent (“log”) culture treated with 0.4 mg P. gingivalis extract/ml for 24 hours (FIG.","1) and 48 hours (FIG.","2).","FIG.","3 and FIG.","4 show confluent culture treated for 24 hours (FIG.","3) and 48 hours (FIG.","4), respectively.","FIG.","1 shows a 70% detachment for active HUVEC culture cells following treatment with a P. gingivalis extract over a 24-hour period.","FIG.","2 shows a 90% detachment of the same cells after 48 hours.","FIG.","3 shows approximately 68% detachment of quiescent Human (HUVEC after 24 hours of treatment, whereas the same cells exhibit nearly 100% detachment after 48 hours (see FIG.","4).","FIG.","5 and FIG.","6 show non-confluent (“log”) culture treated for 24 hours (FIG.","5) and 48 hours (FIG.","6) with 0.4 mg P. gingivalis extract/ml medium.","FIG.","7 and FIG.","8 show confluent cultures treated for 24 hours (FIG.","7) and 48 hours (FIG.","8).","The control is untreated A549.FIG.","5 shows active A549 human non-small cell lung carcinoma exhibited a 95% detachment rate after 24 hours of treatment with a P. gingivalis extract.","After 48 hours, there was no change in the percent detachment of these same cells (see FIG.","6).","FIG.","7 shows 40% detachment rate after 24 hours, whereas after 48 hours the detachment rose to greater than 50%.","Detachment from the plastic surface of the petri dish is consistent with the ability of P. gingivalis extract to degrade β1 integrin.","Anti-angiogenic and tumor-reducing activity correlates with the degree of detachment of the endothelial and cancer tissue culture cells in both 24 and 48-hour treatments.","Demonstrating detachment of up to 96% of the cells (A549) or 9% (HUVEC) can thus be considered pertinent activity.","This is for total protein where the fraction of the active ingredient is small.","Therefore, its activity is very high.","Both exponentially growing (“log”) and stationary phase (“plateau”) quiescent cultures showed differences in ability to remain attached following treatment FIG.","15 total cell number reduction a result of the treatment.","The treatment is more efficient on growing tumor cells than on cells in stationary phase.","Similar reduction was obtained with endothelial (HUVEC) cells.","FIG.","16 demonstrates that detachment of HUVEC is reduced by inhibitors or heating of SPF (soluble protein faction; see Example 9).","In order to determine if the junctional protein-degrading activity is a cysteine proteinase, additional treatments were included Nα-p-tosyl-L-lysine chloromethyl ketone (TLCK), a cysteine proteinase inhibitor, was added at 10 mM.","In addition, heat-inactivation (20 min at 65° C.) of the P. gingivalis extract was performed to inactivate protein-mediated proteolysis.","All experiments were done in triplicate.","Using a Coulter counter, the number of the detached cells was determined and mean values were expressed as percent of the total (detached+attached) cell number.","The results of this experiment demonstrate that a P. gingivalis protein is the active substance and that this protein is a cysteine proteinase.","EXAMPLE 2 Method of Demonstrating Detachment Associated with PrtP Protease from P. gingivalis To demonstrate that protease PrtP isolated from P. gingivalis is responsible for the detachment observed in Example 1, three samples were applied to A549 lung carcinoma cells.","A single P. gingivalis protein, PrtP protease, was expressed in Bacteroides fragilis, a species related to P. gingivalis, but which does not express PrtP, for the purpose of further chromatographic purification.","Treatment of carcinoma cells was performed with an extract of B. fragilis containing PrtP and compared to the same treatment with the wild-type B. fragilis host.","Therefore, the difference between the treatments was limited to the presence/absence of P. gingivalis PrtP protease only.","The strains were grown in BHIS broth (per liter, 37 g Brain Heart Infusion (Difco), 1 g L-Cysteine (Sigma), 5×10−4% hemin, 0.2% NaHCO3 in an anaerobic chamber with an atmosphere of 5% CO2, 10% H2, and 85% N2).","Agar (1.5%) was added for solid medium.","P. gingivalis W83 was grown on Trypticase soy agar (BBL Microbiology stems, Cockeysville, Md.)","supplemented with sheep blood (5%), hemin (5 mg/ml), and menadione (5 mg/ml).","When broth-grown P. gingivalis was required, cultures were grown in Todd-Hewitt broth (BBL Microbiology Systems) supplemented with hemin (5 μg/ml), menadione (5 μg/ml), and glucose 2 mg/ml) anaerobically.","Normal Bacteroides fragilis was used as a control.","Detachment of cells exposed to both B. fragilis with PrtP and P. gingivalis were nearly equal, whereas those cells exposed to the control exhibited minimal detachment.","This study provides direct evidence that PrtP is active in cell proliferation inhibition.","A migration inhibition (“scratch wound”) assay is a method that is routinely used for the ration of the ability of cells to migrate, an important step in tumor neovascularization.","This assay is used by the Biological Testing Branch, Developmental Therapeutics Program at NCI to identify new anti-angiogenic compounds.","Inhibition of migration is considered one aspect of anti-angiogenic activity (Yeh et al., Mol Pharmacol.","2001 May,59(5):1333-42.).","FIG.","9 demonstrates a HUVEC migration inhibition assay: For this assay, endothelial cells were cultured on a slide.","Upon reaching confluency, a 2-mm scrape “wound” was introduced on a central portion of the slide.","The number of cells which migrated into the denuded area was enumerated after further incubation for 24 hours.","The results (the mean of 2 experiments) demonstrate that at 0.4 mg total protein/ml, the migration of the human vascular endothelial cells was reduced by 45%.","EXAMPLE 3 Migration Inhibition Assay To demonstrate that a P. gingivalis extract exerts anti-angiogenic effects, as opposed to general inhibition of cell proliferation, the following assay was performed.","P. gingivalis extracts were produced and homogenized to obtain an extract as described in Example 1.At 0.4 mg total protein/ml, human vascular endothelial cell migration in standard in vitro assay known in the at to reflect angiostatic and anti-tumor acidity, was reduced by 45%, (mean value of 2 experiments).","In addition, at 48 hours, detachment of 85% of log phase lung carcinoma cells was observed (FIG.","10).","Since total cell protein was used, where the faction of the active ingredient is small, this experiment demonstrates that the angiostatic activity of the P. gingivalis proteinase is high.","EXAMPLE 4 Identification of Epithelial Cell Ligands of Hemagglutinin A In order to determine if HagA interacts directly with host cell components, a functionally active fragment of HagA was produced in E. coli using the E. coli expression vector, pET19b (Novagen).","In this system, purification was achieved by fusing a histidine tag to a Hag fragment and by affinity purification of the fusion protein on a Ni2+ column.","Oligonucleotides were designed flanking 2 HArep sequences to include the active site of hemagglutination as disclosed in U.S. Pat.","No.","5,824,791 and to include restriction sites for ligation of the fragment into the expression vector, pET19b.","The 3 kb PCR product was cloned into pT7Blue vector (Novagen), digested with NdeI and XhoL and the coding sequence directionally subcloned into pET19b, which had been digested with the same enzymes and CIP-treated.","Using PCR with a mixed pair of primers T7 (from vector) and ST2/3′ (from insert), transformants in E. coli Novablue (Novagen) were screened for an insert in the proper orientation.","One such clone, pEKS5, was chosen for further work and was transformed into E. coli BL21 (DE3), an expression strain (Novagen).","After induction with 1 mM IPTG, cells were lysed and the lysate was applied to an activated His-Bind resin affinity chromatography column.","Elution with 1 M imidazole-containing buffer produced a single protein species with an apparent molecular mass of ˜100 kDa.","After transfer onto a nitrocellulose membrane, the protein was probed with anti-HagA antibody, 61BG1.3, and its authenticity was confirmed.","The purified recombinant HagA peptide was tested for binding to cell components of two human cell lines using the Far Western Immunoblot.","For this assay, KB oral epithelial cells and human umbilical cord endothelial cells (HUVEC) were grown and lysed in hypotonic buffer containing a cocktail of mammalian proteinase inhibitors.","The cell lysates were loaded on SDS-PAGE gels, transferred to nitrocellulose membranes, blocked with dry fat-free milk in TBS, and overlaid with 0.5 μg/ml of purified recombinant HagA.","After three hours of incubation at ambient temperature followed by washing, the membranes were treated first with anti-HagA Mab and secondly with anti-mouse AP conjugate.","The HagA peptide was found to bind intensely to two proteins, ˜60 kDa and 65 kDa in size, present in both epithelial and endothelial cells.","The HagA peptide also bound to two heavy protein species, >200 kDa, present in endothelial cells.","These results demonstrate that HagA binds to and int with one or more proteins present in host cells and suggest the in vivo existence of a protein complex between HagA and endothelial as well as epithelial proteins.","EXAMPLE 5 Proliferation Inhibition of HUH7 Cells by P. gingivalis Extracts, and by Live P. gingivalis Cells in the Presence/Absence of Inhibitors Freshly collected whole P. gingivalis cells were used for 20-hour treatment at a density of 2×1010 bacteria per ml DMEM (antibiotic-free).","FIG.","11 represents the proliferation inhibition of HUH7 cells by P. gingivalis and E. coli extracts, and by live P. gingivalis cells in the presence/absence of inhibitors (five stars: all cells remain attached, no proliferation inhibition).","L-Cysteine was always present at concentration of 5 mM to stabilize the anti-angiogenic activity.","The proliferation inhibition property of P. gingivalis extract and whole cells is clearly demonstrated on human hepatoma cell line (HUH7).","EXAMPLE 6 Immunofluorescent Microscopy of HUH7 Human Hepatoma Cells Treated with P. gingivalis Extract Tissue culture cells grown in T-75 flasks at 37 C in DMEM (Pen/Strep) in a CO2 incubator were subjected to 8 ml of trypsin-EDTA and incubated at 37 C for 10-15 minutes for detachment.","Trypsinized cells were transferred (with 2×10 ml DMEM) to 50 ml culture tube and centrifuged at 1K rpm for 10 seconds.","Supernatant was removed and cells were washed at 8 ml Ca-free PBS.","Washed cells were centrifuged at K rpm for 10 seconds and excess wash was removed.","Cells were resuspended in 20 ml of DMEM (Pen/Strep) media and transferred to new T-75 flask for incubation at 37 C in 5% CO. HUH7 cells were incubated with P. gingivalis extract (0.8 mg protein per ml of medium) for 20 h at which time the cells were washed three times with phosphate buffered saline (PBS) and then fixed in 4% paraformaldehyde in PBS for 30 minutes at room temperature.","This was followed by washing twice in PBS and quenching in NH4Cl (50 mM)/0.3% Tween 20/PBS for 10 minutes at room temperature.","After quenching, the HUH7 were washed two times in PBS.","The primary antibodies were rabbit anti-human occluding (Zymed Laboratories #71-1500) and rabbit anti-human pan cadherin (Sigma Chemical Co., St Louis, Mo.","#C3678).","They were diluted 1/50 in PBS/5% normal goat serum/0.3% Tween™ 20 and applied to the cells for 2 h at room temperature.","The HUH7 were then washed four times in PBS for 5 minutes each time.","The secondary antibody (rhodamine-conjugated goat anti-rabbit (Sigma)) was applied for 1 h at room temperature.","The HUH7 cells were then washed twice with PBS before mounting with Fluoromount-G (Southern Biotechnology Associates, Inc., Birmingham Ala.) onto glass microscope slides and sealing with nail polish.","Images were viewed using an Olympus IX70 deconvolution microscope and Delatvision software (Applied Precision, Inc., Wepahah, Wash.).","EXAMPLE 7 Reactivity with Anti-Occludin Antibody Occludin-stained junctions degraded upon treatment with P. gingivalis extract.","TLCK presence or heat-inactivation of P. gingivalis extract abolishes the activity.","E. coli extract control treatment exhibited no activity and the occludin network was intact.","FIG.","12A shows non-treated HUH7 cells.","FIG.","12B shows HUH7 cells after treatment with P. gingivalis extract wherein the occludin network was degraded; FIG.","12C shows HUH7 cells after treatment with P. gingivalis extract in the presence of inhibitor TLCK, wherein the occludin network was intact; FIG.","12D shows HUH7 cells following treatment with heat-treated P. gingivalis extract, wherein the occludin network was degraded; and FIG.","12E shows HUH7 cells treated with E. coli extract and demonstrates that E. coli does not effect the occludin network.","The data from immunoflourescent staining or junctional molecules from HUH7 cells confirms the capacity of P. gingivalis extract to disrupt the intracellular network by degrading the border consisting of cell adhesion molecules (CAMs).","EXAMPLE 8 Reactive with Anti-Pan Cadherin Antibody Cadherin-stained junctions were degraded upon treatment with P. gingivalis extract.","TLCK presence or heat-inactivation of P. gingivalis abolish the activity.","FIG.","13A shows control, non-treated HUH7 hepatoma cells, wherein the junctions were intact FIG.","13B shows HUH7 cells following treatment with P. gingivalis extract, wherein the junctions were degraded; FIG.","13C shows HUH7 cells after treatment with P. gingivalis extract in the presence of inhibitor TLCK, wherein the junctions were intact; and FIG.","13D shows HUH7 cells after treatment with heat-inactivated P. gingivalis extract, wherein the junction were intact.","The proliferation as stated above, and the data from immunoflourescent staining of junctional molecules from HUH7 cells confirms the capacity of P. gingivalis extract to disrupt the intracellular network by degrading the border consisting of cell adhesion molecules.","Thus both examples 7 and 8 demodulate that the unique activity of the present disclosed extract can be utilized for tumor disintegration.","EXAMPLE 9 Fractioning of Bacterial Culture Liquor Proteins In order to partially purify the CAM-degrading activity, fractional precipitation of secreted P. gingivalis proteins from spent culture liquor was achieved using ammonium sulfate.","Broth culture grown in an anaerobic chamber was centrifuged for 20 minutes at a speed of 8000 rpm.","Next it was filtered with a 0.2 μm filter (Nalgene) to remove any remaining cells.","To saturate to 60%, 36.1 g of (NH4)2SO4 were dissolved in every 100 ml of culture liquor.","The solution was left stirring overnight at 4 C and collected the next day by centrifuging for 20 minutes at 8000 rpm.","Precipitated proteins were collected from six liters of P. gingivalis W83 spent culture medium.","The protein pellet was resuspended in a 20 ml solution of 50 mM Tris HCl (pH 7.5).","The solution was dialyzed (Pierce SnakeSkin tubing, 7 kDa MWCO) against 50 mM Tris.HCl overnight at 4 C The dialysis was repeated with fresh buffer.","Dialyzed solution was filtered with an Acrodisc® syringe filter (0.2 μm) and then concentrated using a Centriprep 10 (Amicon) for a total of an hour and a half After concentration, the solution was aliquoted, and the protein concentration was determined using the BCA assay (Sigma B-9643).","The solution was then stored at −80 C EXAMPLE 10 Proliferation Inhibition of HUVEC Polarized Cell Line.","HUVEC #Ecv-304 Treatment with P. gingivalis Fractions Polarized endothelial cells cultured on porous membrane inserts (Transwell, Corning Costar Corp., Cambridge, Mass.)","were used as an in vitro model for studying anti-angiogenic activities and to test for differential activity from both sides of the endothelium.","Six hundred id (for 24-well plate) or 2.6 ml (for 6-well plate) of DMEM medium (Penn/Strep) were added to the lower chamber of tissue culture plates.","Vascular endothelial cells were seeded into Corning Costar Transwell inserts in volumes of 0.1 ml medium (24-well plate) or 1.5 ml medium (6-well plate) in the upper chamber.","Cultures were grown to confluence in a CO2 incubator at 37.0 C before being treat The proteins were added to the upper or lower chambers at a final concentration of 0.8 mg/ml.","The cultures were incubated for 4 days in a CO2 incubator.","Similar rets were obtained using whole P. gingivalis cells (data not shown).","FIG.","14A shows control, untreated polarized human endothelial cells ECV-304; FIG.","14B shows polarized ECV-304 cells treated basolaterally with 60% fraction of P. gingivalis culture liquid proteins; and FIG.","14C shows polarized ECV-304 cells following treatment lumenally with 60% fraction of P. gingivalis culture liquid proteins.","In each experiment the polarized endothelial cell layer was treated from either the apical or basolateral side with identical concentrations of protein preparations.","As seen in the optical micrographs in FIGS.","14A, B and C at a point where complete destruction was observed from basolateral application of ammonium sulfate-precipitated proteins, no damage was observed in the cultures with lumenal (apical) application of same preparations.","These data strongly support the conclusions that anti-angiogenic activity is partially purified from P. gingivalis secreted proteins as 60-% fraction of ammonium sulfate-precipitated culture liquor proteins; and the targeting of this activity toward the basolateral, extravascular side of the vasculature is specifically beneficial for degradation of the endothelial vascular cell layer in abnormally leaky tumor vessels.","In addition to anti-angiogenic activity, immunofluorescent and proliferation inhibition studies with human cancer cell lines (hepatoma and lung carcinoma) demonstrate the utility of this P. gingivalis-associated activity for disintegration of extravascular tumor tissues, i.e., direct tumor-disintegration activity exists.","Using the same abnormal openings to access both the basolateral side of the tumor vasculature and the surrounding tumor tissue brings double benefit to the proposed treatment.","In light of foregoing evidence, it is apparent that P. gingivalis polypeptides of the invention can be utilized as a vascular endothelial cell migration inhibitor and as an anti-angiogenic pharmaceutical agent Furthermore, while at present there does not appear to be any known therapeutic protocol based on selective degradation of cell-cell and cell-matrix adhesion molecules in tumors and a large number of other diseases, the present invention provides a new method of disease treatment of such pathologies.","It is further predictable, based on the disclosure provided herein, that other Porphyromonas gingivalis arginine or lysine specific cysteine protease polypeptides, hemagglutinin polypeptides and fragments thereof known or yet to be discovered that exhibit similar anti-angiogenic activity, can be used according to the methods of this invention.","Furthermore, combinations of such molecules can also be used according to the methods of this invention EXAMPLE 11 Effect of P. gingivalis on Integrin Receptors.","α5β1 integrin is upregulated in tumor endothelium (Stupack and Cheresh, Sci STKE.","2002 Feb. 12;2002(119):PE7).","Being an apoptosis regulator, the integrin is a target for anticancer drugs.","B1 integrin is a target of P. gingivalis proteolytic activity in canine epithelial cells (Katz, et al., Infect Immun.","2000 Mar.","68(3):1441-9).","To test if β1 integrin is targeted in a human cell line, immunoanalysis of detached HUVEC was performed after treatment with the P. gingivalis extract to determine the extent of degradation.","Western blots of SDS-PAGE gels were probed using anti-human β1 integrin MAb (BD Transduction Labs, cat.","# MMS-496R).","The results demonstrated that treating with the P. gingivalis extract (except if heat-inactivated) resulted in complete degradation of the 130-kDa integrin.","EXAMPLE 12 Effect of P. gingivalis Proteins on Transendothelial Resistance (TER) of Polarized HUVE Cells Junctional complexes between adjacent polarized endothelial cells constitute a permeability barrier between lumenal and basolateral compartments.","Disruption of the junctional complexes leads to increased flow of solutes, including small ions that can be monitored electrophysiologically by reading the changes in the transendothelial resistance (TER).","HUVE cells were grown onto Transwell inserts with 0.4 μm pore size.","The integrity of the HUVEC layer was decreased by treatment with P. gingivalis SPF compared to mock-treated control See FIG.","17 (the data are mean values from triplicate experiments).","Notably, an identical concentration of SPF applied to the BL (basolateral) compartment decreased TER faster than when applied to the L (lumenal) compartment thus confirming and extending the observations made with canine epithelial cells.","(Katz et al., Infect Immun.","2000 March 68(3):1441-9.)","FIG.","18 shows TER data and micrographs for an extended period of time (8 days).","Similar HUVEC treatment as above was performed in triplicate.","FIG.","19 shows the TER graph FIG.","19 shows contrast micrographs of the cell layers on day 8.FIG.","19A; L-treated; FIG.","19B, BL-treated and FIG.","19C, mock-treated control.","The difference in the TER level reflects the status of the endothelial layer as documented on the micrographs, totally non-existing (BL treatment) and virtually intact (L treatment; control).","Serine proteinase (trypsin) treatment (0.05% in medium) does not provide such specificity.","Using electrophysiology and microscopy on the same cell cultures, these data again demonstrate the destructive action of P. gingivalis secreted proteins on polarized human endothelial cell model and their preference for the BL side of the cell layer.","EXAMPLE 13 Endothelial Degradation Following Treatment with 15-kDa P. gingivalis Protein Cloned in E. coli.","A 15-kDa internal fragment of a HagA repeat (described in Paramaesvaran, et al., 1998, J.","Dental Res., 77:664) was expressed in E. coli.","In this experiment, a confluent endothelial cell layer was treated with a soluble sonic extract from E. coli expressing 15-kDa subunit of the adhesive domain (of gingipains or HagA), 0.5 mg total protein/mL FIG.","20 shows micrographs of endothelial cell layer.","FIG.","20A shows HUVEC treated with control E. coli host extract, 0.5 mg protein/mL FIG.","20C shows control mock-treated cells.","FIG.","20B shows eminent with E. coli expressing 15-kDa protein.","FIG.","21 shows proliferation inhibition of HUVEC treated with 15-kDa P. gingivalis protein cloned in E. coli.","(E. coli 0.5, control E. coli protein extract at 0.5 mg/ml.","E. coli HA2 0.5, E. coli protein extract containing HA2, the 15-kDa polypeptide at 0.5 me).","The results of this experiment demonstrate sharp reduction of cell numbers upon treatment with recombinant 15-kDa P. gingivalis protein.","EXAMPLE 14 P. gingivalis cysteine proteinases and hemagglutinins can mediate detachment of cells from a substrate and inhibit proliferation in human endothelial and carcinoma cells.","Also, junctional molecule degradation and vascular network disintegration using bacterial extracts and whole cells has been demonstrated.","Further, partially purified secreted junctional molecules-targeting proteinase have specificity toward the basolateral side of polarized human endothelial monolayer.","In addition to the endothelial cell experiments, experiments with human cancer cell lines demonstrate the utility of this P. gingivalis-associated activity for treatment of extravascular tumor tissues, i.e.","direct tumor disintegration in addition to anti-angiogenic activity.","This activity can be specifically beneficial for degradation of the endothelial vascular cell layer from the basolateral side in the abnormally leaky tumor vessels.","See FIG.","22.Using same abnormal openings to access both the basolateral side of the tumor vasculature and the extravascular tumor tissue would bring double benefit to the proposed treatment.","FIG.","22 shows potential pathways for leakage from blood vessels in tumors.","Intercellular openings (arrows) between lining cells of a murine tumor vessel viewed by scanning electron microscopy.","Figures A and B show multiple large intercellular openings (arrow, 22A) and three smaller transcellular holes (arrows, 22B) in branched lining cells of a tumor vessel.","The intercellular openings are much larger than the holes.","Region in box in 22A is shown at higher magnification in 22B.","(Hashizume, et al., Am J Pathol.","2000 Apr.","156(4):1363-80.)"]],"string":"[\n [\n \"Methods and compositions for prevention of angioproliferation\",\n \"The invention provides pharmaceutical compositions comprising Porphyromonas gingivalis protease and hemagglutinin polypeptides that have anti-angiogenic activity and methods for their use.\"\n ],\n [\n \"1.A composition for the treatment, prevention, or amelioration of an angioproliferitive condition comprising a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide has anti-angiogenic activity, and a pharmaceutically acceptable excipient.\",\n \"2.The composition of claim.\",\n \"1, wherein the polypeptide is selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.\",\n \"3.The composition of claim 2, wherein the polypeptide has at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, prtT or tla polypeptide and wherein the polypeptide has anti-angiogenic activity.\",\n \"4.The composition of claim 2, wherein the polypeptide is a biologically functional homolog, or isoform of a rgpA, rgpB, kgp, hag, prtT or tla polypeptide.\",\n \"5.The composition of claim 1, wherein the polypeptide is a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or Porphyromonas gingivalis hemagglutinin polypeptide, and wherein the fragment has anti-angiogenic activity.\",\n \"6.The composition of claim 5, wherein the fragment is HA2.7.The composition of claim 1, wherein the angioproliferative condition is carcinoma, sarcoma, melanoma, benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthris, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, arteriosclerosis, scleroderma, hypertrophic scars, cat scratch disease, Helicobacter pylori ulcers, capillary proliferation within atherosclerotic plaque, or a combination thereof.\",\n \"8.A method for the treatment, prevention, or amelioration of an angioproliferative condition comprising administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof wherein the polypeptide has anti-angiogenic activity, to a patient in need thereof whereby the angioproliferative condition is treated, prevented or ameliorated.\",\n \"9.The method of claim 8, wherein the polypeptide is selected from the group co-ng of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.\",\n \"10.The method of claim 8, wherein the polypeptide has at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, or tla polypeptide and wherein the polypeptide has anti-angiogenic activity.\",\n \"11.The method of claim 8, wherein the polypeptide is a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or a Porphyromonas gingivalis hemagglutinin polypeptide, ad wherein the fragment has anti-angiogenic activity.\",\n \"12.The method of claim 11, wherein the polypeptide fragment is HA2.13.The method of claim 8, wherein the angioproliferative condition is carcinoma, sarcoma, melanoma, benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthritis, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, atherosclerosis, scleroderma, hypertrophic scars, cat scratch disease, Helicobacter pylori ulcers, capillary proliferation within atherosclerotic plaque, or a combination thereof.\",\n \"14.The method of claim 8, wherein the method comprises contacting a vasculature supplying a biological structure affected by the angioproliferative condition with the polypeptide.\",\n \"15.The method of claim 14, wherein the polypeptide is contacted with a basolateral surface of the vasculature.\",\n \"16.A method for potentiating effects of a chemotherapeutically effective agent comprising administering to a patient: (a) a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptid, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide disrupts cell-cell adhesion, cell-matrix adhesion, or both; and (b) a chemotherapeutically effective agent, whereby effects of a chemotherapeutically effective agent are potentiated.\",\n \"17.The method of claim 16, wherein the polypeptide is selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.\",\n \"18.The method of claim 17, wherein the polypeptide has at least about 900% sequence identity with a rgpA, rgpB, kgp, hag, or tla polypeptide, and wherein the polypeptide has anti-angiogenic activity.\",\n \"19.The method of claim 16, wherein the polypeptide is a fragment of Porphyromonas gingivalis arginine specific cystine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or Porphyromonas gingivalis hemagglutinin polypeptide, and wherein the fragment has proteinase activity.\",\n \"20.The composition of claim 19, wherein the polypeptide fragment is HA2.21.A method for preventing the formation of new vasculature required for implantation or sustenance of a fertilized an ovum comprising: administering a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a mammal, wherein the polypeptide has anti-angiogenic activity, whereby the formation of new vasculature required for implantation or sustenance of a fertilized mammalian ovum is prevented.\",\n \"22.The method of claim 21, wherein the polypeptide is selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.\",\n \"23.The method of claim 22, wherein the polypeptide has at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, prtT, or ta polypeptide and wherein the polypeptide has anti-angiogenic activity.\",\n \"24.The method of claim 21, wherein the polypeptide is a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or Porphyromonas gingivalis hemagglutinin polypeptide, and wherein the fragment has anti-angiogenic activity.\",\n \"25.The method of claim 24, wherein the polypeptide fragment is HA2.26.A pharmaceutical composition for facilitating passage of a compound through a blood-brain barrier comprising a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, a Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, and a pharmaceutically acceptable excipient.\",\n \"27.The pharmaceutical composition of claim 26 further including a compound to be passed through the blood-brain barrier.\",\n \"28.A method delivering a compound through a blood-brain barrier of a patient comprising administering a pharmaceutically effective amount of a Porphymonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof and the compound, whereby the compound is passed through the blood-brain barrier.\",\n \"29.The method of claim 28, wherein the polypeptide is selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.\",\n \"30.The method of claim 29, wherein the polypeptide has at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, prtT or tla polypeptide and wherein the polypeptide has anti-angiogenic activity.\",\n \"31.The method of claim 28, wherein the polypeptide is a foment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or a Porphyromonas gingivalis hemagglutinin polypeptide, and wherein the fragment has anti-angiogenic activity.\",\n \"32.The method of claim 31, wherein the polypeptide fragment is HA2.33.A method of degrading a tumor comprising: administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a patient having a tumor, wherein the polypeptide degrades cell-cell bonds, cell-matrix bonds, or both cell-cell bonds and cell-matrix bonds of the tumor, whereby the tumor is degraded.\",\n \"34.The method of claim 33 wherein the polypeptide is selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.\",\n \"35.The method of claim 34 wherein the polypeptide has at least about 90% se ce identity with a rgpA, rgpB, kgp, hag, prtT or tla polypeptide and wherein the polypeptide has anti-angiogenic activity.\",\n \"36.The method of claim 33 wherein the polypeptide is a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, or Porphyromonas gingivalis is hemagglutinin polypeptide, and wherein the fragment has anti-angiogenic activity.\",\n \"37.The method of claim 36 wherein the polypeptide fragment is HA2.\"\n ],\n [\n \"<SOH> BACKGROUND INFORMATION <EOH>Cancer is the second leading cause of death in the United States, accounting for over one half million deaths per year.\",\n \"(National Vital Statistics Report, 1998, Vol.\",\n \"48, No.\",\n \"11).\",\n \"The total economic cost associated with cancer has been estimated to be over $100 billion dollars annually.\",\n \"(Brown, M. L. et al.\",\n \"In Cancer Epidemiology and Prevention, 1996).\",\n \"There is currently no cure for the disease, but several lines of research appear promising.\",\n \"Of these, research directed at preventing angiogenesis offers the most hope.\",\n \"Angiogenesis involves a complex biochemical cascade of events that leads to new blood vessel formation in, for example, developing tumors.\",\n \"For cells to survive, each must have some communication, direct or indirect with the existing vasculature in order to obtain nutrients and oxygen, and to offload metabolic waste products.\",\n \"Active vascularization is normally observed following injury to a tissue, during development, or in response to ovulation in females.\",\n \"However, abnormal rapid proliferation of blood vessels is also observed in areas where cancerous masses have developed.\",\n \"Because the rate of cell mass growth is limited by the degree of vascularization present, cancer cells release chemical substances into the surrounding environment to induce nearby, nourishing blood vessels to grow toward the proliferating cancer cell mass.\",\n \"For example, angiopoietin-1 basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF) and other substances are released from cancer cells to coax surrounding blood vessels to grow collateral vessels toward the tumor.\",\n \"Once vascularized, a tumor mass may grow locally, or may metastasize and begin to spread through the bloodstream and lymphatic system to other parts of the body, causing significant damage.\",\n \"For example, vascularization of the retina in diabetic retinopathy can lead to retinal detachment resulting in blindness.\",\n \"Metastasis is a hallmark for malignancy, which in extreme cases may lead to rapid death of an individual.\",\n \"Ironically, some tumors may also secrete angiostatic substances to inhibit tumor growth (Chen et al., Cancer Res.\",\n \"1995, 55, 4230-4233; O'Reilly, Cell 1997, Jan. 24 (88):277-285).\",\n \"Thus, it appears that in healthy individuals angiogenesis associated with tumor growth may be regulated by a fine balance between the release of angiogenic factors and the release of angiostatic factors.\",\n \"It is believed that by blocking the process of angiogenesis, tumor growth can be suspended, which in turn would lead to cancer remission.\",\n \"Most early research directed at preventing angiogenesis involved exposing various cell lines to angiostatic compounds and assessing the degree of proliferation either in vivo or in vitro.\",\n \"The National Cancer Institute, for example, uses proliferation, migration and cord formation assays in HUVE cells for its anti-angiogenesis testing.\",\n \"Several angiostatic agents that function to prevent the proliferation of cancer cells have been isolated and tested.\",\n \"For example, administration of Angiostatin has been shown to suppress vascular endothelial cell proliferation, thereby reducing the size and lethality of tumors (Folman J., Forum Genova 1999 July-Dec. 9 (3 Suppl.\",\n \"3): 5962).\",\n \"Recombinant Endostatin (baculovirus) has been used to inhibit the proliferation of bovine capillary endothelial cells.\",\n \"(O'Reilly, et al., Cell, 1994, Oct. 21; 79(2):185-8).\",\n \"Until recently, angiostatic compounds have included only those substances capable of preventing proliferation of cells.\",\n \"However, a growing body of evidence demonstrates that agents which inhibit proliferation via cellular detachment from tumor masses perform an analogous function.\",\n \"Recent studies have been directed at interfering or disrupting the mechanisms involved in cell-cell or cell-matrix binding as a means to reduce or eliminate cancerous growth.\",\n \"Cells will not proliferate if they are not first attached to a surface.\",\n \"For example, impaired cell-matrix contact leads to anoikis (epithelial apoptosis) (Vitale et al., FEBS Lett.\",\n \"1999 462 (1-2:57-60); Attwell et al., Oncogene 2000 19(33):3811-5; Rosen, J.\",\n \"Cell.\",\n \"Biol 2000 149(2):447-56); Rytomaa et al., Curr.\",\n \"Biol.\",\n \"1999 9(18):1043-6 or endothelial apoptosis (Erdreich-Epstein, Cancer Res.\",\n \"2000 Feb. 1; 60(3):71221).\",\n \"This anchorage dependence is mediated, in part, by cell surface molecules known as integrins.\",\n \"See e.g., Erdreich-Epstein et al., Cancer Res., 2000 Feb. 1; 60(3):712-21; Lee & Juliano, Mol.\",\n \"Biol.\",\n \"Cell, 2000 Jun.\",\n \"11(6):1973-87; Kawahara, J.\",\n \"Cancer Res.\",\n \"Clin.\",\n \"Oncol, 1995 1212 (3):133-40; Lee, Mol.\",\n \"Biol.\",\n \"Cell 2000 11(6):1973-87; Ruoslahti, Kidney Int.\",\n \"1997 51(5):1413-7; Brassard et al., Exp.\",\n \"Cell.\",\n \"Res.\",\n \"1999 251 (1):33-45; Kottke et al., J. Biol.\",\n \"Chem.\",\n \"1999 274(22):15927-36.Anti-angiogenic approaches are the most recent and promising avenue in cancer treatment.\",\n \"Agents capable of blocking vascularization of neoplastic tissue can prevent subsequent growth of transformed tissue and can lead to existing tissue remission.\",\n \"Anti-angiogenic activity has been detected for several endogenous factors.\",\n \"For example, combrestatin A-4 disodium phosphate (CA4DP) (Dark, Cancer Res.\",\n \"1997 May 15; 57 (10):1829-34), a purified human PSA compound (WIPO PCT publication No.\",\n \"WO 99/60984), Endostatin (add, et al., Biochem.\",\n \"Biophys.\",\n \"Res.\",\n \"Commun.\",\n \"1999 Sep. 24:263(2):3405), so (AIbini, Faseb J.\",\n \"1999 13(6):647-55), Epidermal Growth Factors (EGF) (Kotke, et al., J. Bio.\",\n \"Cam 1999 May 28;274(22):15927-36), and angiostatin (Stack, Biochem J., 1999 340 (pt1):77-84) have anti-angiogenic activity.\",\n \"For many years, scientists have been in search of therapeutics that can be used to prevent periodontal diseases, including gum infections and tooth decay.\",\n \"One organism that has been identified as a potential etiologic agent of such pathologies as gingivitis and periodontal disease is the pathogen Porphyromonas gingivalis .\",\n \"Sequences from the pathogen have been cloned and sequenced.\",\n \"Examples of such work can be found in U.S. Pat.\",\n \"Nos.\",\n \"5,824,791 and 5,830,710, both of which are hereby incorporated herein by reference in their entity.\",\n \"Proteolytic enzymes of Porphyromonas gingivalis are the main tool for providing nutrients to these asaccharolytic bacteria and are also important virulence factors.\",\n \"These enzymes have been shown to degrade basement membrane matrix proteins (Uitto, Oral Microbiology and Immunology 1988 3:97-102), (Smalley, Arch.\",\n \"Oral Biol.\",\n \"1988 33 (5):323-9) and purified P. gingivalis cysteine protease has been shown to disrupt the basement membrane of human carcinoma monolayer (Shah, J. Periodontol.\",\n \"1992 63(1):44-51).\",\n \"Targeted disruption of fibronectin-integrin interactions in human gingival fibroblasts has been demonstrated by the RI protease of P. gingivalis W50 (Scragg, et al., Infect.\",\n \"Immun.\",\n \"1999 67(4):1837-43).\",\n \"Other studies have examined the function of P. gingivalis extract on cell-cell and cell-matrix bonds.\",\n \"Cell detachment from each other and from the underlying surface correlates with the cysteine-dependent proteolytic activity of P. gingivalis (Johansson, Eur.\",\n \"J.\",\n \"Oral.\",\n \"Sci.\",\n \"1998 106(4):863-71).\",\n \"B1-integrin, occluding and E-cadherin are targeted by P. gingivalis proteolytic activity in canine epithelial cells (Katz, et al., Infect.\",\n \"Immun.\",\n \"2000 68(3):1441-9).\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>It is an object of the invention to provide compositions and methods for treatment, prevention or amelioration of angioproliferitive conditions.\",\n \"This and other objects of the invention are provided by one or more of the embodiments described below.\",\n \"One embodiment of the invention provides a composition for the t ent, prevention, or amelioration of an angioproliferitive condition.\",\n \"The composition comprises a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide has anti-angiogenic activity, and a pharmaceutically acceptable excipient.\",\n \"The polypeptide can be selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.\",\n \"The polypeptide can have at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, prtT, or tla polypeptide and has anti-angiogenic activity.\",\n \"The polypeptide can be a biologically functional homolog, or isoform of a rgpA, rgpB, kgp, hag, prtT or tla polypeptide.\",\n \"The polypeptide can be a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cystine protease polypeptide, or Porphyromonas gingivalis hemagglutinin polypeptide, such as HA2.Such a fragment has anti-angiogenic activity.\",\n \"An angioproliferative condition can be carcinoma, sarcoma, melanoma, benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthritis, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, arteriosclerosis, scleroderma, hypertrophic scars, cat scratch disease, Helicobacter pylori ulcers, capillary proliferation within atherosclerotic plaque, or a combination thereof.\",\n \"Another embodiment of the invention provides a method for the treatment, prevention, or amelioration of an angioproliferative condition.\",\n \"The method comprises administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a patient in need thereof, wherein the polypeptide has anti-angiogenic activity, whereby the angioproliferative condition is treated, prevented or ameliorated.\",\n \"The method can comprise contacting a vasculature supplying a biological structure affected by the angioproliferative condition with the polypeptide.\",\n \"The polypeptide can be contacted with a basolateral surface of the vasculature.\",\n \"Yet another embodiment of the invention provides a method for potentiating effects of a chemotherapeutically effective agent.\",\n \"The method comprises administering a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide has disrupts cell-cell adhesion, cell-matrix adhesion, or both; and a chemotherapeutically effective agent to a patient.\",\n \"Effects of the chemotherapeutically effective agent arm potentiated.\",\n \"Still another embodiment of the invention provides a method for preventing the formation of now vasculature required for implantation or sustenance of a fertilized mammalian ovum.\",\n \"The method comprises administering a Porphyromonas gingivalis amine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a mammal, wherein the polypeptide has anti-angiogenic activity, whereby the formation of new vasculature required for implantation or sustenance of a fertilized mammalian ovum is prevented.\",\n \"Even another embodiment of the invention provides a pharmaceutical composition for facilitating passage of compounds through a blood-brain barrier comprising a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, a Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, and a pharmaceutically acceptable excipient.\",\n \"The pharmaceutical composition can further include a compound to be passed through the blood-brain barrier.\",\n \"Another embodiment of the invention provides a method of delivering a compound through a blood-brain barrier of a patient comprising administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof and the compound, whereby the compound is passed through the blood-brain barrier.\",\n \"Still another embodiment of the invention provides a method of degrading a tumor.\",\n \"The method comprises administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a patient having a tumor, wherein the polypeptide degrades cell-cell bonds, cell-matrix bonds, or both cell-cell bonds and cell-matrix bonds of the tumor, whereby the tumor is degraded.\",\n \"Therefore, the present invention discloses anti-angiogenic compositions of mater and methods of their use, which are capable of disrupting endothelial tissue growth and proliferation.\",\n \"The methods involve local or systemic application of a P. gingivalis polypeptide to a targeted tissue.\",\n \"The disclosed polypeptides can inhibit angiogenesis associated with malignant tumor proliferation by, for example, disrupting endothelial layer cell-cell and cell-matrix adhesion bonds.\",\n \"Use of these compounds and methods have advantages over conventional treatments such as chemotherapy, because it targets only growing vessels, while leaving intact vessels unaffected.\",\n \"The present invention provides a novel strategy that uses protein, peptide and nucleic acid sequences of Porphyromonas gingivalis to treat or prevent angioproliferative conditions.\"\n ],\n [\n \"PRIORITY This application claims priority to U.S. Ser.\",\n \"No.\",\n \"09/849,115, filed on May 5, 2001, which is incorporated by reference in its entirety herein FIELD OF THE INVENTION The invention provides compositions and methods for the treatment, prevention and amelioration of angioproliferative conditions.\",\n \"BACKGROUND INFORMATION Cancer is the second leading cause of death in the United States, accounting for over one half million deaths per year.\",\n \"(National Vital Statistics Report, 1998, Vol.\",\n \"48, No.\",\n \"11).\",\n \"The total economic cost associated with cancer has been estimated to be over $100 billion dollars annually.\",\n \"(Brown, M. L. et al.\",\n \"In Cancer Epidemiology and Prevention, 1996).\",\n \"There is currently no cure for the disease, but several lines of research appear promising.\",\n \"Of these, research directed at preventing angiogenesis offers the most hope.\",\n \"Angiogenesis involves a complex biochemical cascade of events that leads to new blood vessel formation in, for example, developing tumors.\",\n \"For cells to survive, each must have some communication, direct or indirect with the existing vasculature in order to obtain nutrients and oxygen, and to offload metabolic waste products.\",\n \"Active vascularization is normally observed following injury to a tissue, during development, or in response to ovulation in females.\",\n \"However, abnormal rapid proliferation of blood vessels is also observed in areas where cancerous masses have developed.\",\n \"Because the rate of cell mass growth is limited by the degree of vascularization present, cancer cells release chemical substances into the surrounding environment to induce nearby, nourishing blood vessels to grow toward the proliferating cancer cell mass.\",\n \"For example, angiopoietin-1 basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF) and other substances are released from cancer cells to coax surrounding blood vessels to grow collateral vessels toward the tumor.\",\n \"Once vascularized, a tumor mass may grow locally, or may metastasize and begin to spread through the bloodstream and lymphatic system to other parts of the body, causing significant damage.\",\n \"For example, vascularization of the retina in diabetic retinopathy can lead to retinal detachment resulting in blindness.\",\n \"Metastasis is a hallmark for malignancy, which in extreme cases may lead to rapid death of an individual.\",\n \"Ironically, some tumors may also secrete angiostatic substances to inhibit tumor growth (Chen et al., Cancer Res.\",\n \"1995, 55, 4230-4233; O'Reilly, Cell 1997, Jan. 24 (88):277-285).\",\n \"Thus, it appears that in healthy individuals angiogenesis associated with tumor growth may be regulated by a fine balance between the release of angiogenic factors and the release of angiostatic factors.\",\n \"It is believed that by blocking the process of angiogenesis, tumor growth can be suspended, which in turn would lead to cancer remission.\",\n \"Most early research directed at preventing angiogenesis involved exposing various cell lines to angiostatic compounds and assessing the degree of proliferation either in vivo or in vitro.\",\n \"The National Cancer Institute, for example, uses proliferation, migration and cord formation assays in HUVE cells for its anti-angiogenesis testing.\",\n \"Several angiostatic agents that function to prevent the proliferation of cancer cells have been isolated and tested.\",\n \"For example, administration of Angiostatin has been shown to suppress vascular endothelial cell proliferation, thereby reducing the size and lethality of tumors (Folman J., Forum Genova 1999 July-Dec. 9 (3 Suppl.\",\n \"3): 5962).\",\n \"Recombinant Endostatin (baculovirus) has been used to inhibit the proliferation of bovine capillary endothelial cells.\",\n \"(O'Reilly, et al., Cell, 1994, Oct. 21; 79(2):185-8).\",\n \"Until recently, angiostatic compounds have included only those substances capable of preventing proliferation of cells.\",\n \"However, a growing body of evidence demonstrates that agents which inhibit proliferation via cellular detachment from tumor masses perform an analogous function.\",\n \"Recent studies have been directed at interfering or disrupting the mechanisms involved in cell-cell or cell-matrix binding as a means to reduce or eliminate cancerous growth.\",\n \"Cells will not proliferate if they are not first attached to a surface.\",\n \"For example, impaired cell-matrix contact leads to anoikis (epithelial apoptosis) (Vitale et al., FEBS Lett.\",\n \"1999 462 (1-2:57-60); Attwell et al., Oncogene 2000 19(33):3811-5; Rosen, J.\",\n \"Cell.\",\n \"Biol 2000 149(2):447-56); Rytomaa et al., Curr.\",\n \"Biol.\",\n \"1999 9(18):1043-6 or endothelial apoptosis (Erdreich-Epstein, Cancer Res.\",\n \"2000 Feb. 1; 60(3):71221).\",\n \"This anchorage dependence is mediated, in part, by cell surface molecules known as integrins.\",\n \"See e.g., Erdreich-Epstein et al., Cancer Res., 2000 Feb. 1; 60(3):712-21; Lee & Juliano, Mol.\",\n \"Biol.\",\n \"Cell, 2000 Jun.\",\n \"11(6):1973-87; Kawahara, J.\",\n \"Cancer Res.\",\n \"Clin.\",\n \"Oncol, 1995 1212 (3):133-40; Lee, Mol.\",\n \"Biol.\",\n \"Cell 2000 11(6):1973-87; Ruoslahti, Kidney Int.\",\n \"1997 51(5):1413-7; Brassard et al., Exp.\",\n \"Cell.\",\n \"Res.\",\n \"1999 251 (1):33-45; Kottke et al., J. Biol.\",\n \"Chem.\",\n \"1999 274(22):15927-36.Anti-angiogenic approaches are the most recent and promising avenue in cancer treatment.\",\n \"Agents capable of blocking vascularization of neoplastic tissue can prevent subsequent growth of transformed tissue and can lead to existing tissue remission.\",\n \"Anti-angiogenic activity has been detected for several endogenous factors.\",\n \"For example, combrestatin A-4 disodium phosphate (CA4DP) (Dark, Cancer Res.\",\n \"1997 May 15; 57 (10):1829-34), a purified human PSA compound (WIPO PCT publication No.\",\n \"WO 99/60984), Endostatin (add, et al., Biochem.\",\n \"Biophys.\",\n \"Res.\",\n \"Commun.\",\n \"1999 Sep. 24:263(2):3405), so (AIbini, Faseb J.\",\n \"1999 13(6):647-55), Epidermal Growth Factors (EGF) (Kotke, et al., J. Bio.\",\n \"Cam 1999 May 28;274(22):15927-36), and angiostatin (Stack, Biochem J., 1999 340 (pt1):77-84) have anti-angiogenic activity.\",\n \"For many years, scientists have been in search of therapeutics that can be used to prevent periodontal diseases, including gum infections and tooth decay.\",\n \"One organism that has been identified as a potential etiologic agent of such pathologies as gingivitis and periodontal disease is the pathogen Porphyromonas gingivalis.\",\n \"Sequences from the pathogen have been cloned and sequenced.\",\n \"Examples of such work can be found in U.S. Pat.\",\n \"Nos.\",\n \"5,824,791 and 5,830,710, both of which are hereby incorporated herein by reference in their entity.\",\n \"Proteolytic enzymes of Porphyromonas gingivalis are the main tool for providing nutrients to these asaccharolytic bacteria and are also important virulence factors.\",\n \"These enzymes have been shown to degrade basement membrane matrix proteins (Uitto, Oral Microbiology and Immunology 1988 3:97-102), (Smalley, Arch.\",\n \"Oral Biol.\",\n \"1988 33 (5):323-9) and purified P. gingivalis cysteine protease has been shown to disrupt the basement membrane of human carcinoma monolayer (Shah, J. Periodontol.\",\n \"1992 63(1):44-51).\",\n \"Targeted disruption of fibronectin-integrin interactions in human gingival fibroblasts has been demonstrated by the RI protease of P. gingivalis W50 (Scragg, et al., Infect.\",\n \"Immun.\",\n \"1999 67(4):1837-43).\",\n \"Other studies have examined the function of P. gingivalis extract on cell-cell and cell-matrix bonds.\",\n \"Cell detachment from each other and from the underlying surface correlates with the cysteine-dependent proteolytic activity of P. gingivalis (Johansson, Eur.\",\n \"J.\",\n \"Oral.\",\n \"Sci.\",\n \"1998 106(4):863-71).\",\n \"B1-integrin, occluding and E-cadherin are targeted by P. gingivalis proteolytic activity in canine epithelial cells (Katz, et al., Infect.\",\n \"Immun.\",\n \"2000 68(3):1441-9).\",\n \"SUMMARY OF THE INVENTION It is an object of the invention to provide compositions and methods for treatment, prevention or amelioration of angioproliferitive conditions.\",\n \"This and other objects of the invention are provided by one or more of the embodiments described below.\",\n \"One embodiment of the invention provides a composition for the t ent, prevention, or amelioration of an angioproliferitive condition.\",\n \"The composition comprises a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide has anti-angiogenic activity, and a pharmaceutically acceptable excipient.\",\n \"The polypeptide can be selected from the group consisting of rgpA, rgpB, kgp, hag, prtT and tla polypeptides.\",\n \"The polypeptide can have at least about 90% sequence identity with a rgpA, rgpB, kgp, hag, prtT, or tla polypeptide and has anti-angiogenic activity.\",\n \"The polypeptide can be a biologically functional homolog, or isoform of a rgpA, rgpB, kgp, hag, prtT or tla polypeptide.\",\n \"The polypeptide can be a fragment of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cystine protease polypeptide, or Porphyromonas gingivalis hemagglutinin polypeptide, such as HA2.Such a fragment has anti-angiogenic activity.\",\n \"An angioproliferative condition can be carcinoma, sarcoma, melanoma, benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthritis, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, arteriosclerosis, scleroderma, hypertrophic scars, cat scratch disease, Helicobacter pylori ulcers, capillary proliferation within atherosclerotic plaque, or a combination thereof.\",\n \"Another embodiment of the invention provides a method for the treatment, prevention, or amelioration of an angioproliferative condition.\",\n \"The method comprises administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a patient in need thereof, wherein the polypeptide has anti-angiogenic activity, whereby the angioproliferative condition is treated, prevented or ameliorated.\",\n \"The method can comprise contacting a vasculature supplying a biological structure affected by the angioproliferative condition with the polypeptide.\",\n \"The polypeptide can be contacted with a basolateral surface of the vasculature.\",\n \"Yet another embodiment of the invention provides a method for potentiating effects of a chemotherapeutically effective agent.\",\n \"The method comprises administering a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, wherein the polypeptide has disrupts cell-cell adhesion, cell-matrix adhesion, or both; and a chemotherapeutically effective agent to a patient.\",\n \"Effects of the chemotherapeutically effective agent arm potentiated.\",\n \"Still another embodiment of the invention provides a method for preventing the formation of now vasculature required for implantation or sustenance of a fertilized mammalian ovum.\",\n \"The method comprises administering a Porphyromonas gingivalis amine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a mammal, wherein the polypeptide has anti-angiogenic activity, whereby the formation of new vasculature required for implantation or sustenance of a fertilized mammalian ovum is prevented.\",\n \"Even another embodiment of the invention provides a pharmaceutical composition for facilitating passage of compounds through a blood-brain barrier comprising a pharmaceutically effective amount of a substantially purified Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, a Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof, and a pharmaceutically acceptable excipient.\",\n \"The pharmaceutical composition can further include a compound to be passed through the blood-brain barrier.\",\n \"Another embodiment of the invention provides a method of delivering a compound through a blood-brain barrier of a patient comprising administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof and the compound, whereby the compound is passed through the blood-brain barrier.\",\n \"Still another embodiment of the invention provides a method of degrading a tumor.\",\n \"The method comprises administering a pharmaceutically effective amount of a Porphyromonas gingivalis arginine specific cysteine protease polypeptide, Porphyromonas gingivalis lysine specific cysteine protease polypeptide, Porphyromonas gingivalis hemagglutinin polypeptide, or a combination thereof to a patient having a tumor, wherein the polypeptide degrades cell-cell bonds, cell-matrix bonds, or both cell-cell bonds and cell-matrix bonds of the tumor, whereby the tumor is degraded.\",\n \"Therefore, the present invention discloses anti-angiogenic compositions of mater and methods of their use, which are capable of disrupting endothelial tissue growth and proliferation.\",\n \"The methods involve local or systemic application of a P. gingivalis polypeptide to a targeted tissue.\",\n \"The disclosed polypeptides can inhibit angiogenesis associated with malignant tumor proliferation by, for example, disrupting endothelial layer cell-cell and cell-matrix adhesion bonds.\",\n \"Use of these compounds and methods have advantages over conventional treatments such as chemotherapy, because it targets only growing vessels, while leaving intact vessels unaffected.\",\n \"The present invention provides a novel strategy that uses protein, peptide and nucleic acid sequences of Porphyromonas gingivalis to treat or prevent angioproliferative conditions.\",\n \"BRIEF DESCRIPTION OF THE FIGURES FIG.\",\n \"1 demonstrates percent detachment of active Human Umbilical Vein Endothelial Cells (HUVEC) after 24 hours of treatment with a protein extract from P. gingivalis.\",\n \"Represented are mean values from triplicate experiments.\",\n \"FIG.\",\n \"2 demonstrates percent detachment of active HUVEC after 48 hours of treatment with a protein extract from P. gingivalis.\",\n \"FIG.\",\n \"3 demonstrates percent detachment of quiescent HUVEC after 24 hours of treatment with a protein extract from P. gingivalis.\",\n \"FIG.\",\n \"4 demonstrates percent detachment of quiescent HUVEC after 48 hours of treatment with a protein extract from P. gingivalis.\",\n \"FIG.\",\n \"5 demonstrates percent detachment of active human non-small cell lung carcinoma cell line (A549) after 24 hous of treatment with a protein extract from P. gingivalis.\",\n \"FIG.\",\n \"6 demonstrates percent detachment of active human non-small cell lung carcinoma cell line (A549) after 48 hours of treatment with protein extract from P. gingivalis.\",\n \"FIG.\",\n \"7 demonstrates percent detachment of quiescent human non-small cell lung carcinoma cell line (A549) after 24 hours of treatment with a protein extract from P. gingivalis.\",\n \"FIG.\",\n \"8 demonstrates percent detachment of quiescent human non-small cell lung carcinoma cell line (A549) after 48 hours of treatment with a protein extract from P. gingivalis.\",\n \"FIG.\",\n \"9 demonstrates the reduction in human vascular endothelial cell migration after exposure to a protein extract from P. gingivalis over a 24 hour period.\",\n \"FIG.\",\n \"10 demonstrates detachment of human non-small cell lung carcinoma cell line (A549) after treatment with a proteinase extract of P. gingivalis alone and with an extract of Bacteroides fragilis expressing PrtP protease from P. gingivalis.\",\n \"FIG.\",\n \"11 is a table showing the degree of proliferation inhibition of a HUH7 cell line exposed to P. gingivalis and E. coli extract, and to P. gingivalis cells in the presence or absence of inhibitors.\",\n \"FIG.\",\n \"12A-E shows the results of reactivity of a HUH7 cell line with anti-occludin antibodies.\",\n \"FIG.\",\n \"12A depicts non-treated control HUH7 cells.\",\n \"FIG.\",\n \"12B depicts the HUH7 cells treated with P. gingivalis extract.\",\n \"FIG.\",\n \"12C depicts HUH7 cells treated with P. gingivalis extract in the presence of the inhibitor TLCK.\",\n \"FIG.\",\n \"12D depicts results of treatment with heat-treated P. gingivalis extract.\",\n \"FIG.\",\n \"12E depicts HUH7 cells treated with a control E. coli extract; the occludin network is intact.\",\n \"FIG.\",\n \"13A-D show the result of reactivity of a HUH7 cell tine with an anti-pan cadherin antibody.\",\n \"FIG.\",\n \"13A depicts non-treated control HUH7 cells.\",\n \"FIG.\",\n \"13B depicts HUH7 cells treated with P. gingivalis extract.\",\n \"FIG.\",\n \"13C depicts HUH7 cells treated with P. gingivalis extract in the presence of inhibitor TLCK.\",\n \"FIG.\",\n \"13D depicts HUH7 cells treated with heat inactivated P. gingivalis extract.\",\n \"FIG.\",\n \"14A-C shows proliferation inhibition of a HUVEC polarized cell line.\",\n \"FIG.\",\n \"14A depicts non-treated, control polarized human endothelial cell layer, ECV-304.FIG.\",\n \"14B shows ECV-304 cells treated basolaterally (lower chamber) with a 60% fraction of P. gingivalis culture liquid proteins.\",\n \"FIG.\",\n \"14C depicts the results of ECV-304 cells lumenally treated with a 60% fraction of P. gingivalis culture liquid proteins.\",\n \"FIG.\",\n \"15 demonstrates total cell number reduction as result of treatment with P. gingivalis strain W83 extract.\",\n \"FIG.\",\n \"16 demonstrates that detachment of HUVE cells is reduced by inhibitors or heating of SPF.\",\n \"FIG.\",\n \"17 demonstrates transendothelial resistance of HUVE cells upon treatment with P. gingivalis soluble protein fraction (SPF) over a period of 6 hours.\",\n \"FIG.\",\n \"18 demonstrates transendothelial resistance of HUVE cells upon treatment with P. gingivalis soluble protein fraction (SPF) over a period of 8 days.\",\n \"FIG.\",\n \"19A-C shows contrast micrographs of HUVE cell layers that were treated with P. gingivalis SPF or mock-treated at day 8.FIG.\",\n \"19A shows treated cells.\",\n \"FIG.\",\n \"19B shows BL-treated cells.\",\n \"FIG.\",\n \"19C shows mock-treated control cells.\",\n \"FIG.\",\n \"20A-C show contrast micrographs of HUVE cells treated with extracts of E. coli.\",\n \"FIG.\",\n \"20A shows HUVE cells treated with control E. coli host extract.\",\n \"FIG.\",\n \"20B shows control mock-treated cells.\",\n \"FIG.\",\n \"20C shows cells treated with extracts of E. coli host cells that express a 15 kDa internal fragment of a P. gingivalis HagA repeat.\",\n \"FIG.\",\n \"21 shows proliferation inhibition of HUVE cells treated with HA2 15-kDa P. gingivalis protein cloned in E. coli.\",\n \"(E. coli 0.5: control E. coli protein extract at 0.5 mg/ml.\",\n \"E. coli HA2 0.5: E. coli protein extract containing HA2, the 15-kDa polypeptide at 0.5 mg/ml.\",\n \"FIG.\",\n \"22 shows potential pathways for leakage from blood vessels in tumors.\",\n \"Intercellular openings (arrows) between lining cells of a murine tumor vessel viewed by scanning EM.\",\n \"FIGS.\",\n \"22A and B: Multiple large intercellular openings (arrow, FIG.\",\n \"22A) and three smaller transcellular holes (arrows, FIG.\",\n \"22B) in branched lining cells of a tumor vessel.\",\n \"The intercellular openings are much larger than the holes.\",\n \"The boxed region in FIG.\",\n \"22A is shown at higher magnification in FIG.\",\n \"22B.\",\n \"DETAILED DESCRIPTION OF THE INVENTION “Angiogenesis” or “angioproliferation” means conditions of rapid development of vascular supply to a particular organ or biological site.\",\n \"The rapid development can be uncontrolled and/or pathogenic, as in the development of a tumor.\",\n \"However, angiogenisis or angioproliferation can also be associated with non-pathological conditions such as angiogenesis which occurs upon implantation of a fertilized ova.\",\n \"An anti-angiogenic or anti-angioproliferative composition is capable of reducing or preventing angiogenisis or angioproliferation.\",\n \"The present invention is directed to arginine- or lysine-specific cysteine protease polypeptides and hemagglutinin polypeptides derived from P. gingivalis.\",\n \"The polypeptides can be used to treat or prevent angioproliferative conditions, including but not limited to melanoma, sarcoma, and carcinomas including, for example, breast, colon, lung and prostate carcinomas.\",\n \"Additional pathologies susceptible to treatment according to the present invention include benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthritis, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, scleroderma, hypertrophic scars, cat scratch disease, and Helicobacter pylori ulcer and capillary proliferation within atherosclerotic plaque.\",\n \"Post-surgery adhesions are common complication of gynecologic and abdominal surgery.\",\n \"Such complications can lead to infertility, ectopic pregnancy, chronic pain, prolonged recovery and intestinal obstruction.\",\n \"Adhesions following surgery can be prevented by applying polypeptides of the invention at the end of the operation.\",\n \"Furthermore, the compositions of the invention can be used to control non-pathogenic angiogenic conditions, such as using the compositions as a contraceptive to prevent implantation of fertilized ova.\",\n \"Polypeptides of the Invention Compositions of the invention comprise a substantially purified Porphymonas gingivalis arginine-specific cysteine protease polypeptide, a Porphymonas gingivalis lysine-specific cysteine protease polypeptide, a Porphymonas gingivalis hemagglutinin polypeptide or combinations thereof.\",\n \"These polypeptides are expressed from members of the P. gingivalis protease and hemagglutinin gene family.\",\n \"See e.g., Curtis et al., J. Periodont.\",\n \"Res.\",\n \"34:494 (1999).\",\n \"“Substantially purified” refers to polypeptides that are removed from their natural environment, isolated or separated, and are at least 60% free, preferably 75% free, and most preferably 90% free from other components with which they are naturally associated.\",\n \"Polypeptides of the invention include rgpA arginine-specific cysteine proteases.\",\n \"P. gingivalis rgpA proteases are a group of proteases produced by genes of a homologous loci in different of P. gingivalis.\",\n \"Examples of the rgpA proteases are: prpR1 (Accession number X82680), rgp-1 (Accession number U15282), rgpA (Accession number D26470), prtR (Accession number L26341), prtH (Accession number L27483), and hagE (Accession number AF026946), which is similar to rgp (Accession number A55426).\",\n \"Isoforms of rgpA proteases are also part of the invention and include, for example, RgpA(cat), mt-RgpA(cat) and HRgpA isoforms.\",\n \"See, e.g., Curtis et al., J. Periodont.\",\n \"Res.\",\n \"34:494 (1999).\",\n \"P. gingivalis rgpB arginine-specific proteases are also polypeptides of the invention Examples of rgpB proteases include prtRII (Accession number AF007124), rgp-2 (Accession number U85038), rgpB (Accession number D64081), and prR2 (Rangarajan, Mol Microbiol 1997 Mar.\",\n \"23(5):955-65).\",\n \"The rgpB proteases are all highly related and are derived from homologous genes which occur in different strains of P. gingivalis.\",\n \"Isoforms of rgpB proteases are also part of the invention and include, for example, RgpB and mt-RgpB isoforms.\",\n \"See e.g., Curtis et al., J. Periodont.\",\n \"Res.\",\n \"34:494 (1999).\",\n \"Lysine-specific proteases of P. gingivalis include kgp proteases.\",\n \"kgp proteases include, for example, prtK (Accession number U75366), kgp (Accession numbers U54691; D83258), prtP (Accession numbers U42210; AF017059), kgp(381)-hagD (Accession number U68468).\",\n \"The kgp proteases are all highly related and are derived from homologous genes in different stains of P. gingivalis.\",\n \"Isoforms of kgp proteases are also included in the invention.\",\n \"Hemagglutinin polypeptides of the invention include hagA, hagB, hagC, hagD, hagE, tla (e.g., Accession number Y07618) and prtT (Accession number S75942; M83096).\",\n \"See also, e.g., U.S. Pat.\",\n \"Nos.\",\n \"5,824,791 and 5,830,710 and Aduse-Opoku et al., J. Bacteriol.\",\n \"179:4778-4788 (1997).\",\n \"As disclosed in U.S. Pat.\",\n \"No.\",\n \"5,824,791, a hag gene can be a 7887 bp molecule, which encodes a gene product of 2628 amino acids.\",\n \"Within the gene product, there are four repeat segments: HArep1, HArep2, HArep3 and HArep4.Bach of these segments contains a 15-kDa fragment, HA2, that possesses anti-angiogenic activity and is found in the secreted protein fraction.\",\n \"HA2 is instrumental in production of necrosis, which leads to periodontal disease.\",\n \"HA2 is located within a hagA sequence between amino acids 683 and 819 in the first repeat and is similarly located in the other repeats (see Han et al., Infect.\",\n \"Immun.\",\n \"Page 4002, FIG.\",\n \"2, U.S. Pat.\",\n \"Nos.\",\n \"5,824,791 and 5,830,710).\",\n \"P. gingivalis hemagglutinin polypeptides have cysteine protease activity against arginine containing substrates.\",\n \"Nishikata and Yoshimura, Biochem Bio.\",\n \"Phys.\",\n \"Res.\",\n \"Comm 178:336-40 (1991).\",\n \"P. gingivalis hemagglutinins also function as attachment factors and the substrate binding site is responsible for attachment to erythrocytes.\",\n \"Hemagglutinins also contain “peptidase C25 family” activity.\",\n \"The entire hagA, hagB, hagC, hagD, hagE, prtT or tla polypeptide can be used to produce anti-angiogenic effects.\",\n \"Alternatively, any of the HArep sequences or HA2 can be used Additionally, compounds with similar activity can also be developed which have equal or greater potency than polypeptides of the invention, and such compounds come within the scope of this invention.\",\n \"For example, one of skill in the art can design peptidomimetics using, for example, directed protein evolution or by minimizing the size of the polypeptide so that improved or smaller versions of the polypeptide are developed.\",\n \"Polypeptides of the invention include polypeptides produced from any P. gingivalis rgpA, rgpB, kgp, hag, prtT or tla gene.\",\n \"It is well known in the art that the proteases and hemagglutinins of P. gingivalis are produced from a family of protease and hemagglutinin genes that are highly related.\",\n \"This gene family is known to vary from strain to strain of P. gingivalis.\",\n \"See e.g., Curtis et al., J. Periodont.\",\n \"Res.\",\n \"34:494 (1999).\",\n \"The invention includes all polypeptides produced from the rgpA, rgpB, kgp, hag, prtT or tla gene family that have anti-angiogenic activity.\",\n \"One of skill in the art could determine whether a protease or hemagglutinin gene or polypeptide fell within the rgpA, rgpB, kgp, hag, prtT or tla family by comparing the nucleotide or protein sequence of the gene or protein in question to the known sequences of these genes and proteins.\",\n \"Curtis et al.\",\n \"(J. Peridont.\",\n \"Res.\",\n \"34:464 (1999)) describes these genes families and discloses examples of genes that fall within the P. gingivalis protease and hemagglutinin gene family.\",\n \"U.S. Pat.\",\n \"Nos.\",\n \"5,824,791 and 5,830,710 further describes genes that fall within the hag family.\",\n \"All of these related polypeptides have cysteine protease activity and sequence homology and as such, can be classified into the protease-hemagglutinin family.\",\n \"Polypeptides of the invention can either be full-length polypeptides or fragments of polypeptides.\",\n \"For example, fragments of polypeptides of the invention can comprise about 10, 25, 50, 100, 200, 250, 500, 750, or 1,000 amino acids of polypeptides of the invention Percent sequence identity ha an art recognized meaning and there are a no of methods to measure identity between two polypeptide or polynucleotide sequences.\",\n \"See, e.g., Lesk, Ed, Computational Molecule Biology, Oxford University Press, New York, (1988); Smith, Ed, Biocomputing: Informatics And Genome Projects, Academic Press, New York, (1993); Griffin & Griffin, Eds., Computer Analysis Of Sequence Data, Part I, Humana Press, New Jersey, (1994); von Heinje, Sequence Analysis In Molecular Biology, Academic Press, (1987); and Gribskov & Devereux, Eds., Sequence Analysis Primer, M Stockton Press, New York, (1991).\",\n \"Methods for aligning polynucleotides or polypeptides are codified in computer programs, including the GCG program package (Devereux et al., Nuc.\",\n \"Acids Res.\",\n \"12:387 (1984)), BLASTP, BLASTN, FASTA (Atschul et al., J. Mole Biol.\",\n \"215:403 (1990)), and Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wis. 53711) which uses the local homology algorithm of Smith and Waterman (Adv.\",\n \"App.\",\n \"Math., 2:482-489 (1981)).\",\n \"For example, the computer program ALIGN which employs the FASTA algorithm can be used, with an affine gap search with a gap open penalty of −12 and a gap extension penalty of −2.When using any of the sequence alignment programs to determine whether a particular sequence is, for instance, about 95% identical to a reference sequence, the parameters are set such that the percentage of identity is calculated over the full length of the reference polynucleotide and that gaps in identity of up to 5% of the total number of nucleotides in the reference polynucleotide are allowed.\",\n \"Amino acid sequences having at least about 75, preferably at least about 90, 95, 96, 98, or 99% sequence identity to amino acid sequences of P. gingivalis rgpA, rgpB, kgp, hag, prtT or tla polypeptides and have anti-angiogenic activity are also polypeptides of the invention.\",\n \"Also included in the invention are polypeptides having amino acid sequences which are at least about 75, preferably at least about 90, 95, 96, 98, or 99% sequence identity toamino acid sequences of P. gingivalis rgpA, rgpB, kgp, hag, prtT or tla proteins that are produced by an organism other than P. gingivalis and have anti-angiogenic activity.\",\n \"Polypeptides of the invention also are biologically functional homologs, analogs and isoforms of rgpA, rgpB, kgp, hag, prtT or tla polypeptides.\",\n \"A biologically functional homolog is a polypeptide that has sequence identity to an analogous polypeptide from another species.\",\n \"Homologs can be naturally occurring or can be a polypeptide that does not occur in nature.\",\n \"An isoform is encoded by a distinct mRNA splice variant and can be a naturally occurring polypeptide or can be a polypeptide that does not occur in nature.\",\n \"An analog is a polypeptide having alterations involving one or more amino acid insertions and deletions and/or conservative amino acid substitutions.\",\n \"A homolog, analog or isoform of the invention comprises at least about 75, 90, 95, 96, 98 or 99% sequence identity to a rgpA, rgpB, kgp, hag, prtT or tla polypeptide.\",\n \"A biologically functional homolog, analog or isoform has at least one biological activity of a rgpA, rgpB, kgp, hag, prtT or tla polypeptide.\",\n \"That is, they are capable of preventing endothelial cell proliferation, disruption of vascular endothelium, promoting cellular detachment, inhibiting migration of endothelial cells, blocking formation of new blood vessels, or destroying existing blood vessels feeding, for example, tumors.\",\n \"A biologically functional homolog or analog, isoform, or fragment can be tested for biological activity using, for example a cell proliferation/cell detachment assay (see Example 1), a migration inhibition assay (see Example 2), an occludin-stain junction assay (see Example 7), catherin-stain junction assay (see Example 8) proliferation inhibition of polarized cell assay (see Example 10), degradation of α5β1 integrin assay (see Example 11), or transendothelial resistance assay (see Example 12).\",\n \"A biologically functional homolog, analog, isoform or fragment has about 85%, 90%, 95%, 98%, 99%, 100%, 105%, or 110% biological activity of a specific specie in question.\",\n \"In one embodiment of the invention, a polypeptide of the invention is produced recombinantly.\",\n \"A polynucleotide encoding a polypeptide of the invention can be introduced into a recombinant expression vector, which can be expressed in a suitable expression host cell system using techniques well known in the art.\",\n \"A variety of bacterial, yeast, plant, mammalian, and insect expression systems are available in the art and any such expression system can be used.\",\n \"Optionally, a polynucleotide encoding a polypeptide can be translated in a cell-free translation system.\",\n \"A polypeptide of the invention can also be chemically synthesized.\",\n \"If desired, a polypeptide can be produced as a fusion protein, which can also contain other amino acid sequences, such as amino acid linkers or signal sequences, as well as ligands useful in protein purification, such as glutathione-S-transferase, histidine tag, and staphylococcal protein A.\",\n \"More than one polypeptide of the invention can be present in a fusion protein.\",\n \"Biological Activity of Polypeptides of the Invention Polypeptides of the invention have anti-angiogenic biological activity.\",\n \"That is, they are capable of preventing endothelial cell proliferation, disruption of vascular endothelium, promoting cellular detachment, inhibiting migration of endothelial cells, blocking formation of new blood vessels, or destroying existing blood vessels feeding, for example, tumors or combinations thereof.\",\n \"Therefore, compositions of the invention are capable of inhibiting of tumor enlargement, decreasing tumor mass, and inhabiting, reducing, or preventing related biological processes associated with angiogenesis and vascular supply to a particular biological organ or location, or combinations thereof.\",\n \"In addition, the polypeptides of the invention can be capable of disintegrating cell-to-matrix and cell-to-cell bonds in, for example, a tumor such that the tumor is reduced or eliminated.\",\n \"A polypeptide of the invention has at least one of the above-mentioned biological activities.\",\n \"Polypeptides of the invention can act on cell surface adhesion molecules (CAM's), from the basolateral side of the endothelium.\",\n \"Polypeptides of the invention can inhibit angiogenesis by targeting integrin in cell-cell and cell-matrix adhesion bonds.\",\n \"Of particular importance is the fact that vasculature supplying tumor tissues is aberrantly leaky.\",\n \"Regular blood vessels are well formed and have a well-developed adhesion system to keep them together, while blood vessels supplying tumors are poorly formed and extremely leaky (Herlyn, Immunother.\",\n \"1999 May, 22(3):185); see also, Hashizume, et al., Am.\",\n \"J. Pathol.\",\n \"2000 April 156(4):1363-80).\",\n \"As a result, locally or systemically administered polypeptides of the invention leak out of the vasculature that supplies tumor tissue.\",\n \"Access to the basolateral surface of the vasculature is thereby achieved, which results in disruption of the vascular at that location such that di-ation of the tumor occurs.\",\n \"Therefore, compositions of the invention can disrupt angiogenesis without affecting integrity of normal blood vessels.\",\n \"Methods of Treatment Compositions of the invention can be used to treat several conditions in mammals, including humans.\",\n \"For example, angioproliferative conditions can be treated, prevented or ameliorated according the invention.\",\n \"Methods of treatment comprise administering to a patient having an angioproliferative condition a pharmaceutically effective mount of a composition of the invention such that an anti-angiogenic effect is achieved Angioproliferative conditions include, for example, carcinoma, sarcoma, melanoma, benign tumor, ocular retinopathy, retrolental fibroplasias, psoriasis, angiofibromas, endometriosis, hemangioma, rheumatoid arthritis, Osler Webber Syndrome, myocardial angiogenesis, telangiectasia, hemophiliac joints, wound granulation, intestinal adhesions, post-surgery adhesions, atherosclerosis, scleroderma, hypertrophic scars, cat scratch disease, Helicobacter pylori ulcers, capillary proliferation within atherosclerotic plaque, or a combination thereof.\",\n \"An angioproliferative condition can be selectively treated according to the invention by contacting vasculature supplying a biological structure affected by an angioproliferative condition with an anti-angiogenically effective amount of a composition of the invention.\",\n \"Due to the known leakiness of vasculature supplying, for example, a tumor, local or systemic administration of a composition of this invention facilitates contact with the basolateral surface of said vasculature, including the endothelium, with no effect on normal tissues.\",\n \"Compositions of the invention can also be used to potentiate the effects of a chemotherapeutically effective agent.\",\n \"Such methods comprise co-administering a chemotherapeutically effective agent in the presence of a polypeptide of the invention effective to disrupt cell-cell adhesion, cell-matrix adhesion, or both.\",\n \"Such co-administration can be in the form of a covalent complex, an ionic complex, a mixture simultaneous but separate administration, or administration within a relatively close temp sequence.\",\n \"Appropriate chemotherapeutic agents include, but are not limited to doxorubicin, daunorubicin, doxorubicin, idarubicin, vincristine, 6-mercaptopurine, 6-thioguanine, methotrexate, cytoxan, cytarabine, L-asparaginase, busulfan, cyclophoshamide, melphalan, carmustine, lomustine, 5-fluorouracil methotrexate, fludarabine, bleomycin, docetaxel etoposide, vinorelbine, antibodies, and the like.\",\n \"Polypeptides of the invention can also be used in contraception methods.\",\n \"During pregnancy, the endometrial layer of the uterus becomes thickened and engorged with blood vessels upon implantation of a fertilized ovum.\",\n \"Without a well-developed vasculature the fertilized ovum will not be sustained, and the endometrial layer will be sloughed-off in the form of menses, i.e., menstruation.\",\n \"In one embodiment of the present invention, therapeutic compositions for use as contraceptives are provided.\",\n \"In order to induce contraception, the internal vasulature of the uterus is contacted with a contraceptively effective amount of a polypeptide of the invention.\",\n \"The mode of achieving bioavailability of polypeptides of the invention in this and other angioproliferative conditions can be, for example, through systemic or localized administration, such as intrauterine infusion.\",\n \"Polypeptides of the invention can also be used to facilitate passage of compounds, such as pharmaceuticals, through the blood-brain barer of a mammal.\",\n \"A polypeptide of the invention can be administered along with a compound to permeablize a blood-brain barrier and allow delivery of the compound thought the blood-brain barrier.\",\n \"Such methods comprise co-administering a compound in the presence of a polypeptide of the invention effective to disrupt cell-cell adhesion, cell-matrix adhesion, or both.\",\n \"Such co-administration can be in the form of a covalent complex, an ionic complex, a mixture, simultaneous but separate administration, or administration within a relatively close temporal sequence.\",\n \"Pharmaceutical Compositions and Administration Thereof A pharmaceutically effective amount refers to an amount effective in treating an angiogenic condition in a mammalian patient, such as a human.\",\n \"A pharmaceutically acceptable excipient is a non-toxic carrier, vehicle or adjuvant that can be administered to a patient, together with a polypeptide of this invention, and which does not destroy the pharmacological activity of the polypeptide.\",\n \"Excipients, including carriers, vehicles and adjuvants are well known in the art See, e.g., Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Co., Easton Pa., 1985.A pharmaceutically effective amount is an amount of a polypeptide that achieves a specified functional result.\",\n \"A dosage of a polypeptide of the invention can be determined using routine experimentation Pharmaceutical compositions of the invention can be administered by any number of routes including, but not limited to oral controlled release, intravenous, intramuscular, intra-arterial, intramedullary, intradermal intrathecal intraventricular, transdermal, subcutaneous, intraperitoneal, intranasal parenteral, topical, or rectal means.\",\n \"Pharmaceutical compositions for oral administration can be formulated using pharmaceutically acceptable carriers well known in the art in dosages suitable for oral administration.\",\n \"Such carriers enable the pharmaceutical compositions to be formulated as tablets, pills, dragees, capsules, liquids, gels, sips, slurries, suspensions, and the like, for ingestion by the patient.\",\n \"Pharmaceutical formulations suitable for parenteral administration can be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks' solution, Ringer's solution, or physiologically buffered saline.\",\n \"Aqueous injection suspensions can contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol or dextran.\",\n \"Additionally, suspensions of the active compounds can be prepared as appropriate oily injection suspensions.\",\n \"Suitable lipophilic solvents or vehicles include fatty oils such as sesame oily or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes.\",\n \"Non-lipid polycationic amino polymers also can be used for delivery.\",\n \"Optionally, the Z ion also can contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.\",\n \"For topical or nasal administration, penetrants appropriate to the particular barrier to be permeated are used in the formulation.\",\n \"Such penetrants are generally known in the art.\",\n \"A polypeptide of the invention can also be delivered to a patient by administration of a polynucleotide encoding a polypeptide of the invention.\",\n \"Preferably, injection of a P. gingivalis protease or hemagglutinin polynucleotide, which encodes a polypeptide of the invention, is used to treat, ameliorate, or prevent an angioproliferative condition.\",\n \"In addition to the practical advantages of simplicity of construction and modification, injection of a polynucleotide results in the synthesis of a polypeptide of the invention in the host.\",\n \"The polynucleotide is preferably delivered as “naked DNA” or in a vector.\",\n \"A vector can be a plasmid, such as pBR322, pUC, or ColE1, or an adenovirus vector, such as an adenovirus Type 2 vector or Type 5 vector.\",\n \"Optionally, other vectors can be used, including but not limited to Sindbis virus, simian virus 40, alphavirus vectors, poxvirus vectors, and cytomegalovirus and retroviral vectors, such as murine sarcoma virus, mouse mammary tumor virus, Moloney murine leukemia virus, and Rous sarcoma.\",\n \"Minichromosomes such as MC and MC1, bacteriophages, phagemids, yeast artificial chromosomes, bacterial artificial chromosomes, virus particles, virus-like particles, cosmids (plasmids into which phage lambda cos sites have been inserted) and replicons (genetic elements that are capable of replication under their own control in a cell).\",\n \"A polynucleotide can be, for example, injected intramuscularly to a mammal at a dose of 0.5, 0.75, 1.0, 1.5, 2.0, 2.5, 5 or 10 mg/kg.\",\n \"In vivo delivery of polynucleotides (e.g., plasmid DNA) into results in the cellular uptake and expression of the polynucleotide into a desired polypeptide (Wolff, J.\",\n \"A. et al., Science 247:1465-1468 (1990); Wheeler, C. J. et al, Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci.\",\n \"USA 93:11454-11459 (1996)).\",\n \"The efficiency of in vivo polynucleotide administration can be increased using, for example, chemical agents or physical manipulations.\",\n \"Such chemical agents include cellular toxins such as bupivacaine, cardiotoxin or barium chloride, polymers such as polyvinyl pyrolidone, polyvinyl alcohol polyethyleneimine, polyamidomine, and polyethylene glycol-polyethyleneimine-transferrin complexes that coat the DNA and protect it from DNases and enhance plasmid DNA-based expression or immune responses, particles that interact with the DNA and act as carriers and enhance DNA expression such as nanospheres, microspheres, dendrimers, collagen and polylactide co-glycolides, bulking agents such as sucrose, detergents such as sodium glycocholate, sodium deoxycholate, and beta-cyclodextrin, cationic or non-cationic lipids, DNA binding agents, or agents that enhance plasmid DNA transcription such as histone deacetylase inhibitor FR901228 or S-Bromo-cyclic AMP.\",\n \"Physical manipulations include removal of nerves that control muscle contraction, electroporation, use of intravascular pressure, use of sutures coated with plasmid, use of sponges soaked with DNA as intramuscular depots to prolong DNA delivery, use of special needle-based injection methods, and of needleless-injectors tat propel the DNA into cells.\",\n \"The pharmaceutical compositions of the present invention can be manufactured in a manner that is known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes.\",\n \"The pharmaceutical composition can be provided as a salt and can be formed with many acids, including but not limited to, hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc.\",\n \"Salts tend to be more soluble in aqueous or other protonic solvents than are the corresponding free base forms.\",\n \"In other cases, the preferred preparation can be a lyophilized powder which can contain any or all of the following: 1-50 mM histidine, 0.1%-2% sucrose, and 2-7% mannitol at a pH range of 4.5 to 5.5, that is combined with buffer prior to use.\",\n \"Further details on techniques for formulation and administration can be found in the latest edition of REMINGTON'S PHARMACEUTICAL SCIENCES (Maack Publishing Co., Easton, Pa.).\",\n \"After pharmaceutical compositions have been prepared, they can be placed in an appropriate container and labeled for treatment of an indicated condition.\",\n \"Such labeling would include amount, frequency, and method of administration.\",\n \"Because of the specificity of polypeptides of the invention for rapidly developing vasculature, it is possible to define doses of the polypeptides that are non-toxic to the remainder of a patient, but which nonetheless provide a localized effect to achieve beneficial anti-angiogenic effects.\",\n \"Dosages of a polypeptide of the invention can be about 0.01, 0.1, 1, 2, 5, or 10 mg/kg, of body weight, but one of ordinary skill in the art would recognize that for particular applications, it can be necessary to use either greater or lesser dosages.\",\n \"If a polypeptide of the invention is found to generate an immune response in a patient, processes are known in the art for mimicking biological activity of proteins through development of minienzymes, DNA agents, or other small molecules, repeat administration of active site mimics including for chronic administration.\",\n \"The above disclosure generally describes the present invention, and all patents, patent applications, scientific references cited in this disclosure are expressly incorporated herein.\",\n \"A more complete understanding can be obtained by reference to the following specific examples which are provided for purposes of illusion only and are not intended to limit the scope of the invention.\",\n \"EXAMPLE 1 Methods for Testing Human Endothelial or Carcinoma Cell Detachment Exposed to P. gingivalis Extract To determine the efficacy of P. gingivalis protease extracts in inhibiting cell proliferation, two cell lines were tested.\",\n \"Proliferation inhibition was assessed by determining the detachment of tissue culture cells from their substrate.\",\n \"A549 human non-small cell lung carcinoma cell line (maintained in RPMI-1640 medium (Gibco #11875-135) supplemented with L-Glu and Pen/Strep) and human endothelial cell line (HUVEC, ATCC #ECV-304), which was maintained in M199 medium, supplemented with L-Glutamine (0.05%, Gibco 25030-081), 0.1% Penicillin-Streptomycin (Gibco 15140-122) and 10% PBS (HyClone #SH30071.02, added after heat treatment at 56° C. for 30 min), were used as targets in a cell detachment assay.\",\n \"Exponentially growing (Ex) and quiescent (Plateau) phases of cell culture growth were tested.\",\n \"To obtain extract for activity tests, exponentially grown broth cultures of P. gingivalis strain W83 were pelleted, resuspended in 50 mM HEPES buffer (pH 7.5), and sonicated on ice for 2 min.\",\n \"Following centrifugation at 14,000 rpm at 4 C, the supernatant was filtered through 0.22 μm non-protein-binding filter (Gelman) and stored at −80° C. until needed.\",\n \"Protein concentration was measured using Sigma bicinchoninic acid reagent (#B-9643), BSA standard solution and spectrophotometer Shimadzu UV-1201.Six plates per sample with three plates added for non-treated control were used.\",\n \"104 cells per 60-mm tissue culture plate were used to seed the sample.\",\n \"Detached cells were counted by collecting the medium after treatment (24 or 48 hrs), washing the attached cells with calcium-free phosphate-buffered saline (PBS) and adding the wash to the medium.\",\n \"The pellet was then resuspended in medium to obtain a countable number on the hemocytometer.\",\n \"The 25-square (0.1 μl volume) count ×104 gives the cell count per ml.\",\n \"Two readings were made of each of the three plates.\",\n \"This number represented the detached cell count.\",\n \"The attached cells were trypsinized and collected as above; the plate was washed with PBS and added to the tube.\",\n \"After pelleting and resuspending, two readings were made per each of the three plates to obtain the attached cell count.\",\n \"The mean value of the counts was taken, total number of cells (attached+detached) was obtained and the percentage of detached cells was calculated.\",\n \"Different concentrations of P. gingivalis protein extract were used in the beginning to establish effective concentration to be used throughout.\",\n \"The working concentration was chosen to be 0.4 mg total protein tact per ml cute medium.\",\n \"FIG.\",\n \"1 and FIG.\",\n \"2 show non-confluent (“log”) culture treated with 0.4 mg P. gingivalis extract/ml for 24 hours (FIG.\",\n \"1) and 48 hours (FIG.\",\n \"2).\",\n \"FIG.\",\n \"3 and FIG.\",\n \"4 show confluent culture treated for 24 hours (FIG.\",\n \"3) and 48 hours (FIG.\",\n \"4), respectively.\",\n \"FIG.\",\n \"1 shows a 70% detachment for active HUVEC culture cells following treatment with a P. gingivalis extract over a 24-hour period.\",\n \"FIG.\",\n \"2 shows a 90% detachment of the same cells after 48 hours.\",\n \"FIG.\",\n \"3 shows approximately 68% detachment of quiescent Human (HUVEC after 24 hours of treatment, whereas the same cells exhibit nearly 100% detachment after 48 hours (see FIG.\",\n \"4).\",\n \"FIG.\",\n \"5 and FIG.\",\n \"6 show non-confluent (“log”) culture treated for 24 hours (FIG.\",\n \"5) and 48 hours (FIG.\",\n \"6) with 0.4 mg P. gingivalis extract/ml medium.\",\n \"FIG.\",\n \"7 and FIG.\",\n \"8 show confluent cultures treated for 24 hours (FIG.\",\n \"7) and 48 hours (FIG.\",\n \"8).\",\n \"The control is untreated A549.FIG.\",\n \"5 shows active A549 human non-small cell lung carcinoma exhibited a 95% detachment rate after 24 hours of treatment with a P. gingivalis extract.\",\n \"After 48 hours, there was no change in the percent detachment of these same cells (see FIG.\",\n \"6).\",\n \"FIG.\",\n \"7 shows 40% detachment rate after 24 hours, whereas after 48 hours the detachment rose to greater than 50%.\",\n \"Detachment from the plastic surface of the petri dish is consistent with the ability of P. gingivalis extract to degrade β1 integrin.\",\n \"Anti-angiogenic and tumor-reducing activity correlates with the degree of detachment of the endothelial and cancer tissue culture cells in both 24 and 48-hour treatments.\",\n \"Demonstrating detachment of up to 96% of the cells (A549) or 9% (HUVEC) can thus be considered pertinent activity.\",\n \"This is for total protein where the fraction of the active ingredient is small.\",\n \"Therefore, its activity is very high.\",\n \"Both exponentially growing (“log”) and stationary phase (“plateau”) quiescent cultures showed differences in ability to remain attached following treatment FIG.\",\n \"15 total cell number reduction a result of the treatment.\",\n \"The treatment is more efficient on growing tumor cells than on cells in stationary phase.\",\n \"Similar reduction was obtained with endothelial (HUVEC) cells.\",\n \"FIG.\",\n \"16 demonstrates that detachment of HUVEC is reduced by inhibitors or heating of SPF (soluble protein faction; see Example 9).\",\n \"In order to determine if the junctional protein-degrading activity is a cysteine proteinase, additional treatments were included Nα-p-tosyl-L-lysine chloromethyl ketone (TLCK), a cysteine proteinase inhibitor, was added at 10 mM.\",\n \"In addition, heat-inactivation (20 min at 65° C.) of the P. gingivalis extract was performed to inactivate protein-mediated proteolysis.\",\n \"All experiments were done in triplicate.\",\n \"Using a Coulter counter, the number of the detached cells was determined and mean values were expressed as percent of the total (detached+attached) cell number.\",\n \"The results of this experiment demonstrate that a P. gingivalis protein is the active substance and that this protein is a cysteine proteinase.\",\n \"EXAMPLE 2 Method of Demonstrating Detachment Associated with PrtP Protease from P. gingivalis To demonstrate that protease PrtP isolated from P. gingivalis is responsible for the detachment observed in Example 1, three samples were applied to A549 lung carcinoma cells.\",\n \"A single P. gingivalis protein, PrtP protease, was expressed in Bacteroides fragilis, a species related to P. gingivalis, but which does not express PrtP, for the purpose of further chromatographic purification.\",\n \"Treatment of carcinoma cells was performed with an extract of B. fragilis containing PrtP and compared to the same treatment with the wild-type B. fragilis host.\",\n \"Therefore, the difference between the treatments was limited to the presence/absence of P. gingivalis PrtP protease only.\",\n \"The strains were grown in BHIS broth (per liter, 37 g Brain Heart Infusion (Difco), 1 g L-Cysteine (Sigma), 5×10−4% hemin, 0.2% NaHCO3 in an anaerobic chamber with an atmosphere of 5% CO2, 10% H2, and 85% N2).\",\n \"Agar (1.5%) was added for solid medium.\",\n \"P. gingivalis W83 was grown on Trypticase soy agar (BBL Microbiology stems, Cockeysville, Md.)\",\n \"supplemented with sheep blood (5%), hemin (5 mg/ml), and menadione (5 mg/ml).\",\n \"When broth-grown P. gingivalis was required, cultures were grown in Todd-Hewitt broth (BBL Microbiology Systems) supplemented with hemin (5 μg/ml), menadione (5 μg/ml), and glucose 2 mg/ml) anaerobically.\",\n \"Normal Bacteroides fragilis was used as a control.\",\n \"Detachment of cells exposed to both B. fragilis with PrtP and P. gingivalis were nearly equal, whereas those cells exposed to the control exhibited minimal detachment.\",\n \"This study provides direct evidence that PrtP is active in cell proliferation inhibition.\",\n \"A migration inhibition (“scratch wound”) assay is a method that is routinely used for the ration of the ability of cells to migrate, an important step in tumor neovascularization.\",\n \"This assay is used by the Biological Testing Branch, Developmental Therapeutics Program at NCI to identify new anti-angiogenic compounds.\",\n \"Inhibition of migration is considered one aspect of anti-angiogenic activity (Yeh et al., Mol Pharmacol.\",\n \"2001 May,59(5):1333-42.).\",\n \"FIG.\",\n \"9 demonstrates a HUVEC migration inhibition assay: For this assay, endothelial cells were cultured on a slide.\",\n \"Upon reaching confluency, a 2-mm scrape “wound” was introduced on a central portion of the slide.\",\n \"The number of cells which migrated into the denuded area was enumerated after further incubation for 24 hours.\",\n \"The results (the mean of 2 experiments) demonstrate that at 0.4 mg total protein/ml, the migration of the human vascular endothelial cells was reduced by 45%.\",\n \"EXAMPLE 3 Migration Inhibition Assay To demonstrate that a P. gingivalis extract exerts anti-angiogenic effects, as opposed to general inhibition of cell proliferation, the following assay was performed.\",\n \"P. gingivalis extracts were produced and homogenized to obtain an extract as described in Example 1.At 0.4 mg total protein/ml, human vascular endothelial cell migration in standard in vitro assay known in the at to reflect angiostatic and anti-tumor acidity, was reduced by 45%, (mean value of 2 experiments).\",\n \"In addition, at 48 hours, detachment of 85% of log phase lung carcinoma cells was observed (FIG.\",\n \"10).\",\n \"Since total cell protein was used, where the faction of the active ingredient is small, this experiment demonstrates that the angiostatic activity of the P. gingivalis proteinase is high.\",\n \"EXAMPLE 4 Identification of Epithelial Cell Ligands of Hemagglutinin A In order to determine if HagA interacts directly with host cell components, a functionally active fragment of HagA was produced in E. coli using the E. coli expression vector, pET19b (Novagen).\",\n \"In this system, purification was achieved by fusing a histidine tag to a Hag fragment and by affinity purification of the fusion protein on a Ni2+ column.\",\n \"Oligonucleotides were designed flanking 2 HArep sequences to include the active site of hemagglutination as disclosed in U.S. Pat.\",\n \"No.\",\n \"5,824,791 and to include restriction sites for ligation of the fragment into the expression vector, pET19b.\",\n \"The 3 kb PCR product was cloned into pT7Blue vector (Novagen), digested with NdeI and XhoL and the coding sequence directionally subcloned into pET19b, which had been digested with the same enzymes and CIP-treated.\",\n \"Using PCR with a mixed pair of primers T7 (from vector) and ST2/3′ (from insert), transformants in E. coli Novablue (Novagen) were screened for an insert in the proper orientation.\",\n \"One such clone, pEKS5, was chosen for further work and was transformed into E. coli BL21 (DE3), an expression strain (Novagen).\",\n \"After induction with 1 mM IPTG, cells were lysed and the lysate was applied to an activated His-Bind resin affinity chromatography column.\",\n \"Elution with 1 M imidazole-containing buffer produced a single protein species with an apparent molecular mass of ˜100 kDa.\",\n \"After transfer onto a nitrocellulose membrane, the protein was probed with anti-HagA antibody, 61BG1.3, and its authenticity was confirmed.\",\n \"The purified recombinant HagA peptide was tested for binding to cell components of two human cell lines using the Far Western Immunoblot.\",\n \"For this assay, KB oral epithelial cells and human umbilical cord endothelial cells (HUVEC) were grown and lysed in hypotonic buffer containing a cocktail of mammalian proteinase inhibitors.\",\n \"The cell lysates were loaded on SDS-PAGE gels, transferred to nitrocellulose membranes, blocked with dry fat-free milk in TBS, and overlaid with 0.5 μg/ml of purified recombinant HagA.\",\n \"After three hours of incubation at ambient temperature followed by washing, the membranes were treated first with anti-HagA Mab and secondly with anti-mouse AP conjugate.\",\n \"The HagA peptide was found to bind intensely to two proteins, ˜60 kDa and 65 kDa in size, present in both epithelial and endothelial cells.\",\n \"The HagA peptide also bound to two heavy protein species, >200 kDa, present in endothelial cells.\",\n \"These results demonstrate that HagA binds to and int with one or more proteins present in host cells and suggest the in vivo existence of a protein complex between HagA and endothelial as well as epithelial proteins.\",\n \"EXAMPLE 5 Proliferation Inhibition of HUH7 Cells by P. gingivalis Extracts, and by Live P. gingivalis Cells in the Presence/Absence of Inhibitors Freshly collected whole P. gingivalis cells were used for 20-hour treatment at a density of 2×1010 bacteria per ml DMEM (antibiotic-free).\",\n \"FIG.\",\n \"11 represents the proliferation inhibition of HUH7 cells by P. gingivalis and E. coli extracts, and by live P. gingivalis cells in the presence/absence of inhibitors (five stars: all cells remain attached, no proliferation inhibition).\",\n \"L-Cysteine was always present at concentration of 5 mM to stabilize the anti-angiogenic activity.\",\n \"The proliferation inhibition property of P. gingivalis extract and whole cells is clearly demonstrated on human hepatoma cell line (HUH7).\",\n \"EXAMPLE 6 Immunofluorescent Microscopy of HUH7 Human Hepatoma Cells Treated with P. gingivalis Extract Tissue culture cells grown in T-75 flasks at 37 C in DMEM (Pen/Strep) in a CO2 incubator were subjected to 8 ml of trypsin-EDTA and incubated at 37 C for 10-15 minutes for detachment.\",\n \"Trypsinized cells were transferred (with 2×10 ml DMEM) to 50 ml culture tube and centrifuged at 1K rpm for 10 seconds.\",\n \"Supernatant was removed and cells were washed at 8 ml Ca-free PBS.\",\n \"Washed cells were centrifuged at K rpm for 10 seconds and excess wash was removed.\",\n \"Cells were resuspended in 20 ml of DMEM (Pen/Strep) media and transferred to new T-75 flask for incubation at 37 C in 5% CO. HUH7 cells were incubated with P. gingivalis extract (0.8 mg protein per ml of medium) for 20 h at which time the cells were washed three times with phosphate buffered saline (PBS) and then fixed in 4% paraformaldehyde in PBS for 30 minutes at room temperature.\",\n \"This was followed by washing twice in PBS and quenching in NH4Cl (50 mM)/0.3% Tween 20/PBS for 10 minutes at room temperature.\",\n \"After quenching, the HUH7 were washed two times in PBS.\",\n \"The primary antibodies were rabbit anti-human occluding (Zymed Laboratories #71-1500) and rabbit anti-human pan cadherin (Sigma Chemical Co., St Louis, Mo.\",\n \"#C3678).\",\n \"They were diluted 1/50 in PBS/5% normal goat serum/0.3% Tween™ 20 and applied to the cells for 2 h at room temperature.\",\n \"The HUH7 were then washed four times in PBS for 5 minutes each time.\",\n \"The secondary antibody (rhodamine-conjugated goat anti-rabbit (Sigma)) was applied for 1 h at room temperature.\",\n \"The HUH7 cells were then washed twice with PBS before mounting with Fluoromount-G (Southern Biotechnology Associates, Inc., Birmingham Ala.) onto glass microscope slides and sealing with nail polish.\",\n \"Images were viewed using an Olympus IX70 deconvolution microscope and Delatvision software (Applied Precision, Inc., Wepahah, Wash.).\",\n \"EXAMPLE 7 Reactivity with Anti-Occludin Antibody Occludin-stained junctions degraded upon treatment with P. gingivalis extract.\",\n \"TLCK presence or heat-inactivation of P. gingivalis extract abolishes the activity.\",\n \"E. coli extract control treatment exhibited no activity and the occludin network was intact.\",\n \"FIG.\",\n \"12A shows non-treated HUH7 cells.\",\n \"FIG.\",\n \"12B shows HUH7 cells after treatment with P. gingivalis extract wherein the occludin network was degraded; FIG.\",\n \"12C shows HUH7 cells after treatment with P. gingivalis extract in the presence of inhibitor TLCK, wherein the occludin network was intact; FIG.\",\n \"12D shows HUH7 cells following treatment with heat-treated P. gingivalis extract, wherein the occludin network was degraded; and FIG.\",\n \"12E shows HUH7 cells treated with E. coli extract and demonstrates that E. coli does not effect the occludin network.\",\n \"The data from immunoflourescent staining or junctional molecules from HUH7 cells confirms the capacity of P. gingivalis extract to disrupt the intracellular network by degrading the border consisting of cell adhesion molecules (CAMs).\",\n \"EXAMPLE 8 Reactive with Anti-Pan Cadherin Antibody Cadherin-stained junctions were degraded upon treatment with P. gingivalis extract.\",\n \"TLCK presence or heat-inactivation of P. gingivalis abolish the activity.\",\n \"FIG.\",\n \"13A shows control, non-treated HUH7 hepatoma cells, wherein the junctions were intact FIG.\",\n \"13B shows HUH7 cells following treatment with P. gingivalis extract, wherein the junctions were degraded; FIG.\",\n \"13C shows HUH7 cells after treatment with P. gingivalis extract in the presence of inhibitor TLCK, wherein the junctions were intact; and FIG.\",\n \"13D shows HUH7 cells after treatment with heat-inactivated P. gingivalis extract, wherein the junction were intact.\",\n \"The proliferation as stated above, and the data from immunoflourescent staining of junctional molecules from HUH7 cells confirms the capacity of P. gingivalis extract to disrupt the intracellular network by degrading the border consisting of cell adhesion molecules.\",\n \"Thus both examples 7 and 8 demodulate that the unique activity of the present disclosed extract can be utilized for tumor disintegration.\",\n \"EXAMPLE 9 Fractioning of Bacterial Culture Liquor Proteins In order to partially purify the CAM-degrading activity, fractional precipitation of secreted P. gingivalis proteins from spent culture liquor was achieved using ammonium sulfate.\",\n \"Broth culture grown in an anaerobic chamber was centrifuged for 20 minutes at a speed of 8000 rpm.\",\n \"Next it was filtered with a 0.2 μm filter (Nalgene) to remove any remaining cells.\",\n \"To saturate to 60%, 36.1 g of (NH4)2SO4 were dissolved in every 100 ml of culture liquor.\",\n \"The solution was left stirring overnight at 4 C and collected the next day by centrifuging for 20 minutes at 8000 rpm.\",\n \"Precipitated proteins were collected from six liters of P. gingivalis W83 spent culture medium.\",\n \"The protein pellet was resuspended in a 20 ml solution of 50 mM Tris HCl (pH 7.5).\",\n \"The solution was dialyzed (Pierce SnakeSkin tubing, 7 kDa MWCO) against 50 mM Tris.HCl overnight at 4 C The dialysis was repeated with fresh buffer.\",\n \"Dialyzed solution was filtered with an Acrodisc® syringe filter (0.2 μm) and then concentrated using a Centriprep 10 (Amicon) for a total of an hour and a half After concentration, the solution was aliquoted, and the protein concentration was determined using the BCA assay (Sigma B-9643).\",\n \"The solution was then stored at −80 C EXAMPLE 10 Proliferation Inhibition of HUVEC Polarized Cell Line.\",\n \"HUVEC #Ecv-304 Treatment with P. gingivalis Fractions Polarized endothelial cells cultured on porous membrane inserts (Transwell, Corning Costar Corp., Cambridge, Mass.)\",\n \"were used as an in vitro model for studying anti-angiogenic activities and to test for differential activity from both sides of the endothelium.\",\n \"Six hundred id (for 24-well plate) or 2.6 ml (for 6-well plate) of DMEM medium (Penn/Strep) were added to the lower chamber of tissue culture plates.\",\n \"Vascular endothelial cells were seeded into Corning Costar Transwell inserts in volumes of 0.1 ml medium (24-well plate) or 1.5 ml medium (6-well plate) in the upper chamber.\",\n \"Cultures were grown to confluence in a CO2 incubator at 37.0 C before being treat The proteins were added to the upper or lower chambers at a final concentration of 0.8 mg/ml.\",\n \"The cultures were incubated for 4 days in a CO2 incubator.\",\n \"Similar rets were obtained using whole P. gingivalis cells (data not shown).\",\n \"FIG.\",\n \"14A shows control, untreated polarized human endothelial cells ECV-304; FIG.\",\n \"14B shows polarized ECV-304 cells treated basolaterally with 60% fraction of P. gingivalis culture liquid proteins; and FIG.\",\n \"14C shows polarized ECV-304 cells following treatment lumenally with 60% fraction of P. gingivalis culture liquid proteins.\",\n \"In each experiment the polarized endothelial cell layer was treated from either the apical or basolateral side with identical concentrations of protein preparations.\",\n \"As seen in the optical micrographs in FIGS.\",\n \"14A, B and C at a point where complete destruction was observed from basolateral application of ammonium sulfate-precipitated proteins, no damage was observed in the cultures with lumenal (apical) application of same preparations.\",\n \"These data strongly support the conclusions that anti-angiogenic activity is partially purified from P. gingivalis secreted proteins as 60-% fraction of ammonium sulfate-precipitated culture liquor proteins; and the targeting of this activity toward the basolateral, extravascular side of the vasculature is specifically beneficial for degradation of the endothelial vascular cell layer in abnormally leaky tumor vessels.\",\n \"In addition to anti-angiogenic activity, immunofluorescent and proliferation inhibition studies with human cancer cell lines (hepatoma and lung carcinoma) demonstrate the utility of this P. gingivalis-associated activity for disintegration of extravascular tumor tissues, i.e., direct tumor-disintegration activity exists.\",\n \"Using the same abnormal openings to access both the basolateral side of the tumor vasculature and the surrounding tumor tissue brings double benefit to the proposed treatment.\",\n \"In light of foregoing evidence, it is apparent that P. gingivalis polypeptides of the invention can be utilized as a vascular endothelial cell migration inhibitor and as an anti-angiogenic pharmaceutical agent Furthermore, while at present there does not appear to be any known therapeutic protocol based on selective degradation of cell-cell and cell-matrix adhesion molecules in tumors and a large number of other diseases, the present invention provides a new method of disease treatment of such pathologies.\",\n \"It is further predictable, based on the disclosure provided herein, that other Porphyromonas gingivalis arginine or lysine specific cysteine protease polypeptides, hemagglutinin polypeptides and fragments thereof known or yet to be discovered that exhibit similar anti-angiogenic activity, can be used according to the methods of this invention.\",\n \"Furthermore, combinations of such molecules can also be used according to the methods of this invention EXAMPLE 11 Effect of P. gingivalis on Integrin Receptors.\",\n \"α5β1 integrin is upregulated in tumor endothelium (Stupack and Cheresh, Sci STKE.\",\n \"2002 Feb. 12;2002(119):PE7).\",\n \"Being an apoptosis regulator, the integrin is a target for anticancer drugs.\",\n \"B1 integrin is a target of P. gingivalis proteolytic activity in canine epithelial cells (Katz, et al., Infect Immun.\",\n \"2000 Mar.\",\n \"68(3):1441-9).\",\n \"To test if β1 integrin is targeted in a human cell line, immunoanalysis of detached HUVEC was performed after treatment with the P. gingivalis extract to determine the extent of degradation.\",\n \"Western blots of SDS-PAGE gels were probed using anti-human β1 integrin MAb (BD Transduction Labs, cat.\",\n \"# MMS-496R).\",\n \"The results demonstrated that treating with the P. gingivalis extract (except if heat-inactivated) resulted in complete degradation of the 130-kDa integrin.\",\n \"EXAMPLE 12 Effect of P. gingivalis Proteins on Transendothelial Resistance (TER) of Polarized HUVE Cells Junctional complexes between adjacent polarized endothelial cells constitute a permeability barrier between lumenal and basolateral compartments.\",\n \"Disruption of the junctional complexes leads to increased flow of solutes, including small ions that can be monitored electrophysiologically by reading the changes in the transendothelial resistance (TER).\",\n \"HUVE cells were grown onto Transwell inserts with 0.4 μm pore size.\",\n \"The integrity of the HUVEC layer was decreased by treatment with P. gingivalis SPF compared to mock-treated control See FIG.\",\n \"17 (the data are mean values from triplicate experiments).\",\n \"Notably, an identical concentration of SPF applied to the BL (basolateral) compartment decreased TER faster than when applied to the L (lumenal) compartment thus confirming and extending the observations made with canine epithelial cells.\",\n \"(Katz et al., Infect Immun.\",\n \"2000 March 68(3):1441-9.)\",\n \"FIG.\",\n \"18 shows TER data and micrographs for an extended period of time (8 days).\",\n \"Similar HUVEC treatment as above was performed in triplicate.\",\n \"FIG.\",\n \"19 shows the TER graph FIG.\",\n \"19 shows contrast micrographs of the cell layers on day 8.FIG.\",\n \"19A; L-treated; FIG.\",\n \"19B, BL-treated and FIG.\",\n \"19C, mock-treated control.\",\n \"The difference in the TER level reflects the status of the endothelial layer as documented on the micrographs, totally non-existing (BL treatment) and virtually intact (L treatment; control).\",\n \"Serine proteinase (trypsin) treatment (0.05% in medium) does not provide such specificity.\",\n \"Using electrophysiology and microscopy on the same cell cultures, these data again demonstrate the destructive action of P. gingivalis secreted proteins on polarized human endothelial cell model and their preference for the BL side of the cell layer.\",\n \"EXAMPLE 13 Endothelial Degradation Following Treatment with 15-kDa P. gingivalis Protein Cloned in E. coli.\",\n \"A 15-kDa internal fragment of a HagA repeat (described in Paramaesvaran, et al., 1998, J.\",\n \"Dental Res., 77:664) was expressed in E. coli.\",\n \"In this experiment, a confluent endothelial cell layer was treated with a soluble sonic extract from E. coli expressing 15-kDa subunit of the adhesive domain (of gingipains or HagA), 0.5 mg total protein/mL FIG.\",\n \"20 shows micrographs of endothelial cell layer.\",\n \"FIG.\",\n \"20A shows HUVEC treated with control E. coli host extract, 0.5 mg protein/mL FIG.\",\n \"20C shows control mock-treated cells.\",\n \"FIG.\",\n \"20B shows eminent with E. coli expressing 15-kDa protein.\",\n \"FIG.\",\n \"21 shows proliferation inhibition of HUVEC treated with 15-kDa P. gingivalis protein cloned in E. coli.\",\n \"(E. coli 0.5, control E. coli protein extract at 0.5 mg/ml.\",\n \"E. coli HA2 0.5, E. coli protein extract containing HA2, the 15-kDa polypeptide at 0.5 me).\",\n \"The results of this experiment demonstrate sharp reduction of cell numbers upon treatment with recombinant 15-kDa P. gingivalis protein.\",\n \"EXAMPLE 14 P. gingivalis cysteine proteinases and hemagglutinins can mediate detachment of cells from a substrate and inhibit proliferation in human endothelial and carcinoma cells.\",\n \"Also, junctional molecule degradation and vascular network disintegration using bacterial extracts and whole cells has been demonstrated.\",\n \"Further, partially purified secreted junctional molecules-targeting proteinase have specificity toward the basolateral side of polarized human endothelial monolayer.\",\n \"In addition to the endothelial cell experiments, experiments with human cancer cell lines demonstrate the utility of this P. gingivalis-associated activity for treatment of extravascular tumor tissues, i.e.\",\n \"direct tumor disintegration in addition to anti-angiogenic activity.\",\n \"This activity can be specifically beneficial for degradation of the endothelial vascular cell layer from the basolateral side in the abnormally leaky tumor vessels.\",\n \"See FIG.\",\n \"22.Using same abnormal openings to access both the basolateral side of the tumor vasculature and the extravascular tumor tissue would bring double benefit to the proposed treatment.\",\n \"FIG.\",\n \"22 shows potential pathways for leakage from blood vessels in tumors.\",\n \"Intercellular openings (arrows) between lining cells of a murine tumor vessel viewed by scanning electron microscopy.\",\n \"Figures A and B show multiple large intercellular openings (arrow, 22A) and three smaller transcellular holes (arrows, 22B) in branched lining cells of a tumor vessel.\",\n \"The intercellular openings are much larger than the holes.\",\n \"Region in box in 22A is shown at higher magnification in 22B.\",\n \"(Hashizume, et al., Am J Pathol.\",\n \"2000 Apr.\",\n \"156(4):1363-80.)\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10477007"}}},{"rowIdx":994,"cells":{"text":{"kind":"list like","value":[["Regulating immune response using dendritic cells","Disclosed within a method of altering immune responses using dendritic cells.","One form of the method is a method of inducing immunological tolerance in an individual, where type 2 dendritic cells are administrated to an individual, and where the dendritic cells have been incubated with one or more antigens.","Another form of the method involves altering an immune response, in which liposomes containing where liposomes containing one or more antigens are administrated to an individual, and where the liposomes are modified with surface-bound molecules that target the liposomes to type 2 dendritic cells.","Another form of the method involves reducing immune responsiveness, where liposomes containing one or more antigens are administrated to an individual and where the liposomes are modified with the surface bound molecules that target the liposomes to type 1 dendritic cells or type 2 dendritic cells.","Another form of the method is a method of enhancing immune responsiveness, where liposomes containing one or more antigens are administrated to an individual, and where the liposomes are modified with surface-bound molecules that target the liposomes to mature type 1 dendritic cells."],["1-27.","(canceled) 28.A method of altering an immune response comprising administering to the individual liposomes, wherein the liposomes comprise one or more antigens, wherein the liposomes are modified with surface-bound molecules that target the liposomes to type 2 dendritic cells or mature type 1 dendritic cells.","29.The method of claim 28, wherein the antigens are carbohydrates.","30.The method of claim 28, wherein the antigens are nucleic acids.","31.The method of claim 28, wherein the antigens are peptides.","32.The method of claim 28, wherein the antigens are lipids.","33.The method of claim 28, wherein the antigens are a combination of two or more of the following: carbohydrates, nucleic acids, peptides, or lipids.","34.The method of claim 28, wherein the antigens are MHC, QA, HIV gag, pol, and env (DNA or protein), rheumatoid factor, ICA 89, peripherin, carboxypeptidase H, glutamic acid decarboxylase (GAD), AHNAK, mylein basic protein, retinal S antigen, galactomannoprotein, neuraminidase, influenza matrix protein M1, CMV env proteins pp60, ds DNA, thyroglobulin, insulin, pancreatic islet beta cell antigens, MART-1 (melanoma protein), tyrosinase-related protein 2 (TRP2), melanoma cell lysate, MAGE-A3, allogeneic cell lysate, HLA antigens (protein or DNA), CD31 (protein or DNA), aspergillus chitin, pancreatic carcinoma cell line Panc-1 lysate, ErbB-2/neu, human epithelial cell mucin (Muc-1), Ha-ras oncogene product, p53, carcino-embryonic antigen (CEA), raf oncogene product, gp100/pmel17, GD2, GD3, GM2, TF, sTn, MAGE-1, MAGE-3, BAGE, GAGE, tyrosinase, gp75, Melan-A/Mart-1, gp100, HER2/neu, EBV-LMP 1 & 2, HPV-F4, 6, 7, prostate-specific antigen (PSA), prostate specific membrane antigen (PSMA), HPV-16, MUM, alpha-fetoprotein (AFP), CO17-IA, GA733, gp72, p53, ras oncogene product, HPV E7, Wilm's tumor antigen-1, telomerase, melanoma gangliosides, malignant B cell antigen receptor, malignant B cell immunoglobulin idiotype, variable region of an immunoglobulin, hypervariable region or complementarity determining region (CDR) of a variable region of an immunoglobulin, malignant T cell receptor (TCR), variable region of a TCR, hypervariable region of a TCR, or a combination.","35.The method of claim 28, wherein the surface-bound molecule is a monoclonal antibody.","36.The method of claim 28, wherein the surface-bound molecule is a peptide high-affinity ligand.","37.The method of claim 28, wherein the surface-bound molecule is specific for CD11c+, BDCA-1, or both, wherein mature type 1 dendritic cells are targeted.","38.The method of claim 37, wherein the liposomes enhance anti-tumor immune responses.","39.The method of claim 37, wherein the liposomes enhance anti-cancer immune responses.","40.The method of claim 28, wherein the surface-bound molecule is specific for CD123, BDCA-2, BDCA-4, or a combination, wherein type 2 dendritic cells are targeted.","41.The method of claim 40, wherein the liposomes reduce an auto-immune response.","42.The method of claim 41, wherein the auto-immune response is involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and systemic lupus erythematosis.","43.The method of claim 40, wherein the liposomes reduce an allergic immune response.","44.The method of claim 43, wherein the allergic immune response is involved in asthma, gluten allergy, and atopic dermatitis.","45.The method of claim 40, wherein the liposomes reduce graft rejection immune response.","46.The method of claim 45, wherein the individual is an allograft recipient.","47.The method of claim 45, wherein the graft rejection immune response comprises complications associated with graft rejection.","48.The method of claim 45, wherein the graft rejection immune response comprises graft rejection.","49.The method of claim 40, wherein the individual is a hematopoietic stem cell recipient.","50.The method of claim 40, wherein the liposomes reduce graft verses host immune response.","51.The method of claim 40, wherein the antigens are auto-immune antigens.","52.The method of claim 51, wherein the auto-immune antigens are involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, or systemic lupus erythematosis.","53.The method of claim 28, wherein the antigens are antigens involved in allograft rejection, wherein the liposomes reduce the allograft rejection.","54.The method of claim 28, wherein the antigens are antigens involved in graft rejection of hematopoietic stem cell transplants, wherein the liposomes reduce the graft rejection.","55.The method of claim 28, wherein the antigens are antigens involved in graft verses host disease.","56.A method of reducing an immune response comprising administering to the individual liposomes, wherein the liposomes comprise one or more antigens, wherein the liposomes are modified with surface-bound molecules that target the liposomes to type 2 dendritic cells.","57.The method of claim 28, wherein the liposomes enhance an immune response, wherein the liposomes are targeted to immature type 1 dendritic cells.","58.The method of claim 28, wherein the liposomes reduce an immune response, wherein the liposomes are targeted to type 2 dendritic cells.","59.The method of claim 58, wherein the type 2 dendritic cells are immature type 2 dendritic cells.","60.A method of enhancing an immune response comprising administering to the individual liposomes, wherein the liposomes comprise one or more antigens, wherein the liposomes are modified with surface-bound molecules that target the liposomes to mature type 1 dendritic cells.","61.A method of modifying liposomes comprising packaging one or more antigens into a lipopsome, wherein the liposome is modified with surface-bound molecules that target the liposome to type 2 dendritic cells or mature type 1 dendritic cells."],["<SOH> BACKGROUND OF THE INVENTION <EOH>Dendritic cells have specialized capacity to present peptide antigens to T-cells and regulate the initiation of the immune response (Hart, D. N. (1997) “Dendritic cells: unique leukocyte populations which control the primary immune response.” Blood 90: 3245-3287; Bancereau, J. and R. M. Steinman (1998) “Dendritic cells and the control of immunity.” Nature 392: 245-252).","There is significant heterogeneity within the dendritic cell lineage.","Dendritic cells develop from hematopoietic progenitor “stem” cells under the influence of cytokines which act during early myelo-monocytic differentiation (Peters, J. H., J. Ruppert, et al.","(1991) “Differentiation of human monocytes into CD14 negative accessory cells: do dendritic cells derive from the monocytic lineage?” Pathobiology 59: 122-126;Santiago, S. F., E. Belilos, et al.","(1992) “TNF in combination with GM-CSF enhances the differentiation of neonatal cord blood stem cells into dendritic cells and macrophages.” J Leukoc Biol 52(3): 274-81; Grouard, G., M. C. Rissoan, et al.","(1997) “The enigmatic plasmacytoid T cells develop into dendritic cells with interleukin (IL)-3 and CD40-ligand.” J Exp Med 185: 1101-11; Olweus, J., A. BitMansour, et al.","(1997) “Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.” Proc Natl Acad Sci USA 94: 12551-6; Rissoan, M. C., V. Soumelis, et al.","(1999) “Reciprocal control of T helper cell and dendritic cell differentiation.” Science 283: 1183-6; Siegal, F. P., N. Kadowaki, et al.","(1999) “The nature of the principal type 1 interferon-producing cells in human blood.” Science 284: 1835-7; Bergthiier, R., C. Martinon-Ego, et al.","(2000) “A two step-culture method starting with early growth factors permits enhanced production of functional dendritic cells from murine splenocytes.” Immunological Methods 239: 95-107; Ferlazzo, G., J. Klein, et al.","(2000) “Dendritic cells generated from CD34+ progenitor cells with flt3 ligand, c-kit ligand, GM-CSF, IL4, and TNF-alpha are functional antigen-presenting cells resembling monocyte-derived dendritic cells.” Journal of Immunotherapy 23: 48-58).","Phenotypic and functional analyses have demonstrated two general types of dendritic cells, type 1 (DC 1) and type 2 (DC2), that differ in surface marker expression as well as their functional effect on cognate T-cells.","Studies of cytokines used to mobilize peripheral blood hematopoietic progenitors have indicated increased numbers of DC I after administration of Flt3 and GM-CSF, while increased numbers of DC2 have been noted after treatment with G-CSF (Arpinati, M., C. L. Green, et al.","(2000) “Granulocyte-colony stimulating factor mobilizes T helper-2-inducing dendritic cells.” Blood 95: 2484-2490).","Interaction of DC1 with T-cells results in polarization of the T-cells towards Th1 and Tc 1 immune responses characterized by the generation of cytotoxic effectors and production of IL12, TNF and IFN-gamma.","In contrast, interaction of DC2 with T-cells leads to T-cells that facilitate humoral immune responses, secrete IL4 and IL4, and suppress cytotoxic Th1 immune responses.","DC have an important role in the anti-tumor effect of autologous and allogeneic transplantation.","Antigen primed DC in have been used with promising results in adoptive vaccination against tumors (Mayordomo, J. I., T. Zorina, et al.","(1995) “Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity.” Nature Medicine 1(12): 1297-302; Reichardt, V. L., C. Y. Okada, et al.","(1999) “Idiotype vaccination using dendritic cells after autologous peripheral blood stem cell transplantation for multiple myeloma—a feasibility study.” Blood 93(7): 2411-9).","DC1 promote Th1 immune responses in responding CD4 + T-cells characterized by enhanced INF-gamma, TNF, and IL-12 synthesis.","DC2 promote Th2 responses in CD4+ T-cells characterized by IL4 and IL10 synthesis, and inhibition of INF-gamma and TNF production in cognate T-cells (Rissoan, M. C., V. Soumelis, et al.","(1999) “Reciprocal control of T helper cell and dendritic cell differentiation.” Science 283: 1183-6).","Bone marrow contains monocytes and CD86 + , CD34 + progenitor cells which can differentiate into DC1 in the presence of TNF and GM-CSF (Sallusto, F. and A. Lanzavecchia (1994) “Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor a.” Journal of Experimental Medicine 179: 1109-1118; Ye, Z., A. P. Gee, et al.","(1996) “In vitro expansion and characterization of dendritic cells derived from human bone marrow CD34+ cells.” Bone Marrow Transplantation 18(5): 997-1008; Ryncarz, R. E. and C. Anasetti (1998) “Expression of CD86 on human marrow CD34(+) cells identifies immunocompetent committed precursors of macrophages and dendritic cells.” Blood 91(10): 3892-900), as well as CD123 bright DC progenitors which differentiate into DC2 in the presence of IL3 (Olweus, J., A. BitMansour, et al.","(1997) “Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.” Proc Natl Acad Sci U S A 94: 12551-6).","Most studies involving hematopoietic progenitor cell transplantation to date have examined the role of DC1 as a method of adoptive immunotherapy for cancer following autologous HPC transplantation (Mayordomo, J. I., T. Zorina, et al.","(1995) “Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity.” Nature Medicine 1(12): 1297-302).","One recent report, using a murine transplant model, indicated that only host CD11c+DC (DC 1) were necessary for the development of acute GvHD (Shlomchik, W. D., M. S. Couzens, et al.","(1999) “Prevention of graft versus host disease by inactivation of host antigen-presenting cells.” Science 285: 412-5).","Donor DC did not appear to efficiently present host antigens via cross-priming in a way that led to the initiation of GvHD (Shlomchik, W. D., M. S. Couzens, et al.","(1999) “Prevention of graft versus host disease by inactivation of host antigen-presenting cells.” Science 285: 412-5).","In this study, the possible effect of donor DC on inhibiting GvHD was not examined.","The potential role of increased numbers of donor DC2 in regulating GvHD after G-CSF mobilized allogeneic peripheral stem cell transplants has recently been recognized (Arpinati, M., C. L. Green, et al.","(2000).","“Granulocyte-colony stimulating factor mobilizes T helper-2-inducing dendritic cells.” Blood 95: 2484-2490)."],["<SOH> BRIEF SUMMARY OF THE INVENTION <EOH>In accordance with the purposes of this invention, as embodied and broadly described herein, this invention, in one aspect, relates to a method of inducing immunological tolerance in an individual, where the method comprises administering to the individual type 2 dendritic cells or immature type 1 dendritic cells, where the dendritic cells have been incubated with one or more antigens.","In another aspect, the invention relates to a method of reducing immune responsiveness to particular antigens, where the method comprises administering to the individual liposomes, where the liposomes contain one or more antigens, and where the liposomes are modified with surface-bound molecules that target the liposomes to either type 2 dendritic cells or mature type 1 dendritic cells.","In another aspect, the invention relates to a method of enhancing immune responsiveness, where the method comprises administering to the individual liposomes, where the liposomes contain one or more antigens, where the liposomes are modified with surface-bound molecules that target the liposomes to mature type I dendritic cells.","In these various aspects, the antigens can be, for example, carbohydrates, nucleic acids, peptides, lipids, or a combination of two or more of carbohydrates, nucleic acids, peptides, and lipids.","In particular, the antigens can be MHC, QA, HIV gag, pol, and env (DNA or protein), rheumatoid factor, ICA 89, peripherin, carboxypeptidase H, glutamic acid decarboxylase (GAD), AHNAK, mylein basic protein, retinal S antigen, galactomannoprotein, neuraminidase, influenza matrix protein M1, CMV env proteins pp60, ds DNA, thyroglobulin, insulin, pancreatic islet beta cell antigens, MART-1 (melanoma protein), tyrosinase-related protein 2 (TRP2), melanoma cell lysate, MAGE-A3, allogeneic cell lysate, HLA antigens (protein or DNA), CD31 (protein or DNA), aspergillus chitin , pancreatic carcinoma cell line Panc-1 lysate, ErbB-2/neu, human epithelial cell mucin (Muc-1), Ha-ras oncogene product, p53, carcino-embryonic antigen (CEA), raf oncogene product, gp100/pmel17, GD2, GD3, GM2, TF, sTn, MAGE-1, MAGE-3, BAGE, GAGE, tyrosinase, gp75, Melan-A/Mart-1, gp100, HER2/neu, EBV-LMP 1 & 2, HPV-F4, 6, 7, prostate-specific antigen (PSA), prostate specific membrane antigen (PSMA), HPV-16, MUM, alpha-fetoprotein (AFP), CO17-1A, GA733, gp72, p53, ras oncogene product, HPV E7, Wilm's tumor antigen-1, telomerase, melanoma gangliosides, malignant B cell antigen receptor, malignant B cell immunoglobulin idiotype, variable region of an immunoglobulin, hypervariable region or complementarity determining region (CDR) of a variable region of an immunoglobulin, malignant T cell receptor (TCR), variable region of a TCR, hypervariable region of a TCR, or a combination.","The antigens also can be auto-immune antigens, such as antigens involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.","The antigens can be antigens involved in allograft rejection, such that the induced tolerance reduces the allograft rejection.","The antigens also can be antigens involved in graft rejection of hematopoietic stem cell transplants, such that the induced tolerance reduces the graft rejection.","The antigens also can be antigens involved in graft verses host disease.","For those aspects of the disclosed method involving induction of immunological tolerance, the immunological tolerance can reduce an auto-immune response, such as an auto-immune response involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.","The immunological tolerance can reduce an allergic immune response, such as an allergic immune response is involved in asthma, gluten allergy, and atopic dermatitis.","The graft rejection immune response can comprise graft rejection and/or complications associated with graft rejection.","The immunological tolerance can reduce graft verses host immune response or graft rejection immune response.","The individual can be an allograft recipient, such as a hematopoietic stem cell recipient.","The disclosed method of administration of antigen incubated dendritic cells can have additional features and aspects.","For example, the incubation step can occur or be performed ex vivo, and type 2 dendritic cells can be isolated prior to the incubation by selection with an anti-BDCA-2 antibody.","In the case of type 2 dendritic cells, the dendritic cells can be further purified selection for CD123+ cells and/or removal of CD11c− cells.","Thus, in one aspect of the disclosed method, the type 2 dendritic cells are isolated prior to the incubation with antigen by selection with an anti-BDCA-2 antibody, selection for CD 123+ cells, and removal of CD11c− cells.","Alternatively, type 1 dendritic cells can be isolated with an anti-BDCA-1 antibody.","Type 1 dendritic cells can be further purified by removal of BDCA-2+ and/or CD123+ cells.","The disclosed method of administration of liposomes targeted to dendritic cells can have additional features and aspects.","For example, the surface-bound molecule can be a monoclonal antibody, or a peptide high-affinity ligand.","In some embodiments, the surface-bound molecules can be specific for CD11c+, BDCA-1, or both.","This targets mature type 1 dendritic cells In other embodiments, the surface-bound molecules can be specific for CD123, BDCA-2, BDCA-4, or a combination.","This targets type 2 dendritic cells.","The disclosed method can have a variety of effects and objects.","For example, the liposomes can enhance anti-tumor immune responses, can enhance anti-cancer immune responses, can reduce an auto-immune response.","In the latter case, the auto-immune response can be involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.","The liposomes can reduce an allergic immune response, such as an allergic immune response is involved in asthma, gluten allergy, and atopic dermatitis.","The liposomes can reduce graft verses host immune responses in individuals who are hematopoietic allograft recipients, such as hematopoietic stem cell transplant recipients.","The liposomes can reduce graft rejection immune responses such as those that are complications associated with organ allograft transplants.","In another aspect, the invention relates to a method of modifying liposomes, where the method comprises packaging one or more antigens into a lipopsome, where the liposome is modified with surface-bound molecules that target the liposome to either type 2 dendritic cells or immature type 1 dendritic cells.","Additional advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.","The advantages of the invention will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims.","It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed."],["CROSS-REFERENCE TO RELATED APPLICATIONS This application claims benefit of U.S.","Provisional Application No.","60/289,625, filed May 8, 2001, which application is hereby incorporated herein by reference in its entirety.","FIELD OF THE INVENTION The disclosed invention is generally in the field of altering immune responses and specifically in the field of altering immune responses via dendritic cells.","BACKGROUND OF THE INVENTION Dendritic cells have specialized capacity to present peptide antigens to T-cells and regulate the initiation of the immune response (Hart, D. N. (1997) “Dendritic cells: unique leukocyte populations which control the primary immune response.” Blood 90: 3245-3287; Bancereau, J. and R. M. Steinman (1998) “Dendritic cells and the control of immunity.” Nature 392: 245-252).","There is significant heterogeneity within the dendritic cell lineage.","Dendritic cells develop from hematopoietic progenitor “stem” cells under the influence of cytokines which act during early myelo-monocytic differentiation (Peters, J. H., J. Ruppert, et al.","(1991) “Differentiation of human monocytes into CD14 negative accessory cells: do dendritic cells derive from the monocytic lineage?” Pathobiology 59: 122-126;Santiago, S. F., E. Belilos, et al.","(1992) “TNF in combination with GM-CSF enhances the differentiation of neonatal cord blood stem cells into dendritic cells and macrophages.” J Leukoc Biol 52(3): 274-81; Grouard, G., M. C. Rissoan, et al.","(1997) “The enigmatic plasmacytoid T cells develop into dendritic cells with interleukin (IL)-3 and CD40-ligand.” J Exp Med 185: 1101-11; Olweus, J., A. BitMansour, et al.","(1997) “Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.” Proc Natl Acad Sci USA 94: 12551-6; Rissoan, M. C., V. Soumelis, et al.","(1999) “Reciprocal control of T helper cell and dendritic cell differentiation.” Science 283: 1183-6; Siegal, F. P., N. Kadowaki, et al.","(1999) “The nature of the principal type 1 interferon-producing cells in human blood.” Science 284: 1835-7; Bergthiier, R., C. Martinon-Ego, et al.","(2000) “A two step-culture method starting with early growth factors permits enhanced production of functional dendritic cells from murine splenocytes.” Immunological Methods 239: 95-107; Ferlazzo, G., J. Klein, et al.","(2000) “Dendritic cells generated from CD34+ progenitor cells with flt3 ligand, c-kit ligand, GM-CSF, IL4, and TNF-alpha are functional antigen-presenting cells resembling monocyte-derived dendritic cells.” Journal of Immunotherapy 23: 48-58).","Phenotypic and functional analyses have demonstrated two general types of dendritic cells, type 1 (DC 1) and type 2 (DC2), that differ in surface marker expression as well as their functional effect on cognate T-cells.","Studies of cytokines used to mobilize peripheral blood hematopoietic progenitors have indicated increased numbers of DC I after administration of Flt3 and GM-CSF, while increased numbers of DC2 have been noted after treatment with G-CSF (Arpinati, M., C. L. Green, et al.","(2000) “Granulocyte-colony stimulating factor mobilizes T helper-2-inducing dendritic cells.” Blood 95: 2484-2490).","Interaction of DC1 with T-cells results in polarization of the T-cells towards Th1 and Tc 1 immune responses characterized by the generation of cytotoxic effectors and production of IL12, TNF and IFN-gamma.","In contrast, interaction of DC2 with T-cells leads to T-cells that facilitate humoral immune responses, secrete IL4 and IL4, and suppress cytotoxic Th1 immune responses.","DC have an important role in the anti-tumor effect of autologous and allogeneic transplantation.","Antigen primed DC in have been used with promising results in adoptive vaccination against tumors (Mayordomo, J. I., T. Zorina, et al.","(1995) “Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity.” Nature Medicine 1(12): 1297-302; Reichardt, V. L., C. Y. Okada, et al.","(1999) “Idiotype vaccination using dendritic cells after autologous peripheral blood stem cell transplantation for multiple myeloma—a feasibility study.” Blood 93(7): 2411-9).","DC1 promote Th1 immune responses in responding CD4+ T-cells characterized by enhanced INF-gamma, TNF, and IL-12 synthesis.","DC2 promote Th2 responses in CD4+ T-cells characterized by IL4 and IL10 synthesis, and inhibition of INF-gamma and TNF production in cognate T-cells (Rissoan, M. C., V. Soumelis, et al.","(1999) “Reciprocal control of T helper cell and dendritic cell differentiation.” Science 283: 1183-6).","Bone marrow contains monocytes and CD86+, CD34+ progenitor cells which can differentiate into DC1 in the presence of TNF and GM-CSF (Sallusto, F. and A. Lanzavecchia (1994) “Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor a.” Journal of Experimental Medicine 179: 1109-1118; Ye, Z., A. P. Gee, et al.","(1996) “In vitro expansion and characterization of dendritic cells derived from human bone marrow CD34+ cells.” Bone Marrow Transplantation 18(5): 997-1008; Ryncarz, R. E. and C. Anasetti (1998) “Expression of CD86 on human marrow CD34(+) cells identifies immunocompetent committed precursors of macrophages and dendritic cells.” Blood 91(10): 3892-900), as well as CD123brightDC progenitors which differentiate into DC2 in the presence of IL3 (Olweus, J., A. BitMansour, et al.","(1997) “Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.” Proc Natl Acad Sci U S A 94: 12551-6).","Most studies involving hematopoietic progenitor cell transplantation to date have examined the role of DC1 as a method of adoptive immunotherapy for cancer following autologous HPC transplantation (Mayordomo, J. I., T. Zorina, et al.","(1995) “Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity.” Nature Medicine 1(12): 1297-302).","One recent report, using a murine transplant model, indicated that only host CD11c+DC (DC 1) were necessary for the development of acute GvHD (Shlomchik, W. D., M. S. Couzens, et al.","(1999) “Prevention of graft versus host disease by inactivation of host antigen-presenting cells.” Science 285: 412-5).","Donor DC did not appear to efficiently present host antigens via cross-priming in a way that led to the initiation of GvHD (Shlomchik, W. D., M. S. Couzens, et al.","(1999) “Prevention of graft versus host disease by inactivation of host antigen-presenting cells.” Science 285: 412-5).","In this study, the possible effect of donor DC on inhibiting GvHD was not examined.","The potential role of increased numbers of donor DC2 in regulating GvHD after G-CSF mobilized allogeneic peripheral stem cell transplants has recently been recognized (Arpinati, M., C. L. Green, et al.","(2000).","“Granulocyte-colony stimulating factor mobilizes T helper-2-inducing dendritic cells.”Blood 95: 2484-2490).","BRIEF SUMMARY OF THE INVENTION In accordance with the purposes of this invention, as embodied and broadly described herein, this invention, in one aspect, relates to a method of inducing immunological tolerance in an individual, where the method comprises administering to the individual type 2 dendritic cells or immature type 1 dendritic cells, where the dendritic cells have been incubated with one or more antigens.","In another aspect, the invention relates to a method of reducing immune responsiveness to particular antigens, where the method comprises administering to the individual liposomes, where the liposomes contain one or more antigens, and where the liposomes are modified with surface-bound molecules that target the liposomes to either type 2 dendritic cells or mature type 1 dendritic cells.","In another aspect, the invention relates to a method of enhancing immune responsiveness, where the method comprises administering to the individual liposomes, where the liposomes contain one or more antigens, where the liposomes are modified with surface-bound molecules that target the liposomes to mature type I dendritic cells.","In these various aspects, the antigens can be, for example, carbohydrates, nucleic acids, peptides, lipids, or a combination of two or more of carbohydrates, nucleic acids, peptides, and lipids.","In particular, the antigens can be MHC, QA, HIV gag, pol, and env (DNA or protein), rheumatoid factor, ICA 89, peripherin, carboxypeptidase H, glutamic acid decarboxylase (GAD), AHNAK, mylein basic protein, retinal S antigen, galactomannoprotein, neuraminidase, influenza matrix protein M1, CMV env proteins pp60, ds DNA, thyroglobulin, insulin, pancreatic islet beta cell antigens, MART-1 (melanoma protein), tyrosinase-related protein 2 (TRP2), melanoma cell lysate, MAGE-A3, allogeneic cell lysate, HLA antigens (protein or DNA), CD31 (protein or DNA), aspergillus chitin, pancreatic carcinoma cell line Panc-1 lysate, ErbB-2/neu, human epithelial cell mucin (Muc-1), Ha-ras oncogene product, p53, carcino-embryonic antigen (CEA), raf oncogene product, gp100/pmel17, GD2, GD3, GM2, TF, sTn, MAGE-1, MAGE-3, BAGE, GAGE, tyrosinase, gp75, Melan-A/Mart-1, gp100, HER2/neu, EBV-LMP 1 & 2, HPV-F4, 6, 7, prostate-specific antigen (PSA), prostate specific membrane antigen (PSMA), HPV-16, MUM, alpha-fetoprotein (AFP), CO17-1A, GA733, gp72, p53, ras oncogene product, HPV E7, Wilm's tumor antigen-1, telomerase, melanoma gangliosides, malignant B cell antigen receptor, malignant B cell immunoglobulin idiotype, variable region of an immunoglobulin, hypervariable region or complementarity determining region (CDR) of a variable region of an immunoglobulin, malignant T cell receptor (TCR), variable region of a TCR, hypervariable region of a TCR, or a combination.","The antigens also can be auto-immune antigens, such as antigens involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.","The antigens can be antigens involved in allograft rejection, such that the induced tolerance reduces the allograft rejection.","The antigens also can be antigens involved in graft rejection of hematopoietic stem cell transplants, such that the induced tolerance reduces the graft rejection.","The antigens also can be antigens involved in graft verses host disease.","For those aspects of the disclosed method involving induction of immunological tolerance, the immunological tolerance can reduce an auto-immune response, such as an auto-immune response involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.","The immunological tolerance can reduce an allergic immune response, such as an allergic immune response is involved in asthma, gluten allergy, and atopic dermatitis.","The graft rejection immune response can comprise graft rejection and/or complications associated with graft rejection.","The immunological tolerance can reduce graft verses host immune response or graft rejection immune response.","The individual can be an allograft recipient, such as a hematopoietic stem cell recipient.","The disclosed method of administration of antigen incubated dendritic cells can have additional features and aspects.","For example, the incubation step can occur or be performed ex vivo, and type 2 dendritic cells can be isolated prior to the incubation by selection with an anti-BDCA-2 antibody.","In the case of type 2 dendritic cells, the dendritic cells can be further purified selection for CD123+ cells and/or removal of CD11c− cells.","Thus, in one aspect of the disclosed method, the type 2 dendritic cells are isolated prior to the incubation with antigen by selection with an anti-BDCA-2 antibody, selection for CD 123+ cells, and removal of CD11c− cells.","Alternatively, type 1 dendritic cells can be isolated with an anti-BDCA-1 antibody.","Type 1 dendritic cells can be further purified by removal of BDCA-2+ and/or CD123+ cells.","The disclosed method of administration of liposomes targeted to dendritic cells can have additional features and aspects.","For example, the surface-bound molecule can be a monoclonal antibody, or a peptide high-affinity ligand.","In some embodiments, the surface-bound molecules can be specific for CD11c+, BDCA-1, or both.","This targets mature type 1 dendritic cells In other embodiments, the surface-bound molecules can be specific for CD123, BDCA-2, BDCA-4, or a combination.","This targets type 2 dendritic cells.","The disclosed method can have a variety of effects and objects.","For example, the liposomes can enhance anti-tumor immune responses, can enhance anti-cancer immune responses, can reduce an auto-immune response.","In the latter case, the auto-immune response can be involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.","The liposomes can reduce an allergic immune response, such as an allergic immune response is involved in asthma, gluten allergy, and atopic dermatitis.","The liposomes can reduce graft verses host immune responses in individuals who are hematopoietic allograft recipients, such as hematopoietic stem cell transplant recipients.","The liposomes can reduce graft rejection immune responses such as those that are complications associated with organ allograft transplants.","In another aspect, the invention relates to a method of modifying liposomes, where the method comprises packaging one or more antigens into a lipopsome, where the liposome is modified with surface-bound molecules that target the liposome to either type 2 dendritic cells or immature type 1 dendritic cells.","Additional advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.","The advantages of the invention will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims.","It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is a graph of different cells and treatment versus counts per minute (cpm; representing proliferation of the cells).","As described in the Example, 50,000 DC were cultured with 50,000 T-cells in the presence of allo-antigen or 1 μg/ml Con A.","FIG.","2 is a graph of responder to stimulator ratio versus counts per minute (cpm; representing cell proliferation).","As described in the Example, serial dilutions of purified DC were prepared with 50,000, 25,000, 12,500, 6,250, 3125, 1500, or 75 cells added to culture wells in combination with 50,000 T-cells.","DETAILED DESCRIPTION OF THE INVENTION The present invention may be understood more readily by reference to the following detailed description of preferred embodiments of the invention and the Example included therein and to the Figures and their previous and following description.","It is to be understood that this invention is not limited to specific synthetic methods, specific preparative and analytical techniques unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary.","It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.","Immature DC may be immuno-suppressive, and decrease antigen specific immune responses.","A recently published study in normal donors evaluated the ability of immature dendritic cells to augment T-cell activation after in vitro exposure to antigen (Dhodapkar, M. V., C. Y. Li, et al.","(1994) “Clinical spectrum of clonal proliferations of T-large granular lymphocytes: a T-cell clonopathy of undetermined significance?” Blood 84(5): 1620-7).","Immature dendritic cells were derived from peripheral blood monocytes and then exposed to the influenza peptide MP, and KLH.","After return of these DCs to the host, a marked decline in the frequency of MP specific INF-gamma producing cells was noted, and persisted for 30 days.","At the same time, the frequency of influenza specific T-cells was unchanged suggesting that the decrease in MP specific INF producing cells was related to DC exposure to the antigen, rather than a global reduction in influenza response.","Also, this reduction in MP specific INF-gamma producing cells was paralleled by an increase in the frequency of IL-10 producing MP specific cells, and a reduction in the ability of CD8+ Mp specific cells to kill target cells that had been loaded with Mp peptides.","There was no reduction in the overall frequency of MP specific T-cells after return of the immature DC's to account for this change in antigen response.","Recently, a new paradigm has been developed in allogeneic hematopoietic cell transplantation, namely, that donor DC2 inhibit chronic GvHD and GvT responses among recipients of allogeneic HPC transplants.","Based upon the naturally occurring biologic variation of the DC2 content in bone marrow grafts from 113 normal allogeneic sibling donors, it was found that recipients of larger numbers of DC2 experienced more relapse, while recipients of fewer DC2 had less relapse and more chronic GvHD post-transplant (Waller, E. K., H. Rosenthal, et al.","(2001) “Larger numbers of CD4(bright) dendritic cells in donor bone marrow are associated with increased relapse after allogeneic bone marrow transplantation.” Blood 97(10): 2948-56).","Patients received un-manipulated bone marrow from HLA-matched siblings for the treatment of lymphoma, leukemia, and multiple myeloma.","The quantity of CD3+ and CD34+ cells and DC2 content in the bone marrow grafts were measured by flow cytometry and correlated with clinical outcomes post-transplant.","When the study was initiated, a rare population of CD3−, CD4bright, CD8−, low side scatter, bone marrow cells was noted.","These cells were clearly distinct from CD4+ T-cells and CD4lo monocytes.","The CD3−, CD4bright are identical to DC2 (CD123+, CD11c−) subsequently described in peripheral blood, lymph node, thymus, and bone marrow (Grouard, G., M. C. Rissoan, et al.","(1997) “The enigmatic plasmacytoid T cells develop into dendritic cells with interleukin (IL)-3 and CD40-ligand.” J Exp Med 185: 1101-11; Olweus, J., A. BitMansour, et al.","(1997) “Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.” Proc Natl Acad Sci USA 94: 12551-6).","For these and other reasons, it was realized that dendritic cells could be used to alter immune responses using the disclosed method.","Further, it was realized that functional differences between DC1 and DC2 can be exploited using the disclosed method.","A. Definitions As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.","Thus, for example, reference to “a liposome” includes mixtures of two or more such liposomes, and the like.","Ranges may be expressed herein as from “about” one particular value, and/or to “about” another particular value.","When such a range is expressed, another embodimentincludes from the one particular value and/or to the other particular value.","Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment.","It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint.","In this specification and in the claims which follow, reference will be made to a number of terms which shall be defined to have the following meanings: “Optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.","“Dendritic cell” or “DC”, as used herein, refers to a mature antigen presenting cell,which is identified by the expression of one or more of the following markers on its cell surface: CD40, CD80, CD86, CD83 and HLA-DR, or a dendritic cell progenitor, as defined below, or both.","“Dendritic cell progenitor”, “dendritic progenitor cells”, “dendritic precursor cell”, “dendritic cell precursor”, “DCP”, or “immature dendritic cell”, used interchangeably herein, means a hematopoietic cell that differentiates intro a mature DC.","Type 1 Dendritic cell progenitors are typically identified by the expression of one or more of the following markers on its cell surface: CD11c, CD13, CD14, CD33, CD34, HLA_DR, or CD4.Type 2 Dendritic cell progenitors are typically identified by the expression of one or more of the following markers on its cell surface: CD123, CD45RA, CD34, CD36, BDCA-2, BDCA-4, HLA-DR or CD4.Dendritic cell progenitors typically lack expression of co-stimulatory molecules CD40, CD80, and CD86 as well as the activation marker CD83.Dendritic cell progenitors also lack expression of CD3, CD56, and CD20, markers associated with T-cell, NK cell, or B-cell lineages.","“Mature dendritic cell or DC”, as used herein, refers to a mature antigen presenting cell, which is identified by the expression of one or more of the following markers on its cell surface CD54, CD40, CD80, CD86, CD83 and HLA-DR. “Type 1 dendritic cells or DC1” refer to mature dendritic cells or dendritic cell precursors that express CD11c and CD1b but lack expression of BDCA-2.“Type 2 dendritic cells or DC2” refer to mature dendritic cells or dendritic cell precursors that lack expression of CD11c and lack expression of CD1b but express high levels of CD123 and BDCA-2.“Hematopoietic system reconstituting cells” means a population of cells, preferably human, that possess the capability of dividing and producing progeny that include all of the formed cellular elements of the blood.","Sources of hematopoietic system reconstituting cells can include bone marrow (both fetal and adult) and peripheral blood mononuclear cells (PBMC).","“Donor” or “donor source” means the animal, preferably human, that is the naturalsource from which cells are originally removed.","“Recipient” means the animal, typically human, into which cells or liposomes are transplanted.","“Allogeneic” means that the recipient is not the natural source from which transplant cells have been removed.","B.","Components and Compositions Disclosed are the components to be used to prepare the disclosed compositions as well as the compositions themselves to be used within the methods disclosed herein.","These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc.","of these materials are disclosed that, while specific reference of each various individual and collective combinations and permutation of these compounds may not be explicitly disclosed, each is specifically contemplated and described herein.","For example, if a particular liposome is disclosed and discussed and a number of modifications that can be made to a number of molecules including the liposome are discussed, specifically contemplated is each and every combination and permutation of liposomes and the modifications that are possible unless specifically indicated to the contrary.","Thus, if a class of liposomes A, B, and C are disclosed as well as a class of antigens D, E, and F, and an example of a combination molecule (that is, for example, an antigen-loaded liposome) A-D is disclosed, then even if each is not individually recited, each combination is individually and collectively contemplated.","This means, in this example, that combinations, A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F should be considered disclosed.","Likewise, any subset or combination of these is also disclosed.","Thus, for example, the sub-group of A-E, B-F, and C-E would be considered disclosed.","This concept applies to all aspects of this application including, but not limited to, steps in methods of making and using the disclosed compositions.","Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the disclosed methods.","C. Dendritic Cells Dendritic cells (DC) are antigen presenting cells which are important in the initiation of immune responses.","Based on in vitro culturing and characterization, DC are thought to represent a family of related cell types, perhaps from a single lineage.","Progenitors of DC (DCP) can now be identified in many tissues, such as bone marrow and blood, based on the expression of certain cell surface markers.","One type of precursor cell, the Langerhans cell, is found in the epidermis.","Work on Langerhans cells established that these immature precursors are capable of antigen uptake, but do not demonstrate a significant ability to stimulate T cells until they differentiate into mature DC.","A variety of markers have been identified that are useful in defining certain cell types of the dendritic cell family.","For example, markers expressed on the surface of mature dendritic cells can include CD1a, CD11c, DEC-205 (mouse), CD4, CD33, CD208 (DC-LAMP), CD54, CD11b.","HLA-DR, CD40, CD80, CD83, and CD86.Typically, the following markers are not found on the surface of mature dendritic cells: CD20, CD3, CD14, CD16, CD2, CD207 (Langerin), E-cadherin, Birbeck granules, CD56, and CD19.Cell surface markers present on type-I dendritic cell precursors include CD11c, CD13, CD14, CD33, CD34, and HLA-DR.","Cell surface markers present on type-2 dendritic cell precursors include CD4, CD45RA, CD34, CD123, HLA-DR, and CD36, while the following markers are not found on the surface of dendritic cell precursors: CD20, CD3, CD16, CD2, CD40, CD80, CD83 and CD86 (see for example Caux et al.","Intl.","Immunology 1994 6(8): 1177-1185; Steptoe et al.","J.","Immunology 1997 159:5483-5491; Grouard et al.","(J. Exp.","Med.","1997 185(6): 1101-1111; O'Doherty et al.","Immunology 1994 82: 487-493; Galy et al., J. Immunotherapy 1998 21(2): 132-141; Strunk et al., J. Exp.","Med.","1997 185(6): 1131-1136; Olweus et al.","Proc.","Natl.","Acad.","Sci.","USA 1997 94:12551-12556; Prignano et al., Intl.","J. Dermatol.","1998 37:116-119).","By the appropriate use of these markers, dendritic cells can be distinguished from dendritic cell progenitors.","In particular, dendritic cells are mature antigen presenting cells, which can be identified by the expression of one or more of the following markers on its cell surface: CD40, CD80, CD86, CD83 and HLA-DR. Further, unless the context indicates otherwise, a dendritic cell progenitor is a “dendritic cell.” Dendritic cell progenitors are hematopoietic cells which can be identified by the expression of one or more of the following markers on its cell surface: Type-1 dendritic cell progenitors—CD11c, CD13, CD14, CD33, CD34, and HLA-DR. Type-2 dendritic cell progenitors—CD4, CD45RA, CD34, CD123, HLA-DR, and CD36.1.Type 2 Dendritic Cells Type 2 Dendritic cells or “plasmacytoid” dendritic cells are characterized as hematopoietic cells that can present antigen in the context of both MHC class I and MHC class II.","These cells are CD11b−, CD11c−, CD123bright, BDCA-2+, BDCA-4+, HLA-DR+, lineage-(CD3−, CD14−, CD19−, and CD56−).","In addition to presenting antigen to T cells type 2 dendritic cells also secrete IL-4 which suppresses IL-12 secretion by T-cells and redirects T-cell immune responses from a TH1 response to TH2.Type 2 dendritic cells are largely responsible for presenting allergens to T cells.","2.Type 1 Dendritic Cells Type 1 Dendritic cells are characterized as hematopoietic cells that can present antigen in the context of both MHC class I and MHC class II.","These cells are CD11b+, CD11c+, CD123dim, BDCA-1+, HLA-DR+, lineage-(CD3−, CD14−, CD19−, and CD56−).","In addition to presenting antigen to T cells upon antigen encounter and co-stimulation through CD40-CD40L interactions, type 1 dendritic cells also up regulate IL-12 and secrete IFN-γ which suppresses direct T-cell immune responses towards a TH1 response.","Type 1 dendritic cells are largely responsible for presenting bacteria, viruses, and intracellular parasites to T cells.","3.Isolation of Dendritic Cells Dendritic cells to be incubated with antigens can be isolated using any suitable techniques.","Useful techniques involve selection or removal of cells based on the presence of cell surface markers.","Many cells, including dendritic cells useful for the disclosed method, carry and lack unique sets of cell surface markers.","Useful markers identifying dendritic cells are described above and elsewhere herein.","Any one or a combination of these markers can be used in the isolation of dendritic cells.","Particularly useful for the disclosed method are the markers CD11c, CD123 and BDCA-2.The presence of CD123 and BDCA-2, and the absence of CD11c identify a particularly useful dendritic cell subset: type 2 dendritic cells (DC2).","Cells can be selected for retention via markers present on the desired cells.","Cells can be selected for removal via markers not present on the desired cells.","Thus, the cells remaining after cells are removed via a particular cell marker are enriched for cells lacking that cell marker.","Both selection and removal of cells using cell markers can be accomplished using, for example, specific binding molecules targeted to the cell marker involved.","Useful specific binding molecules for cell separation are antibodies direct to and/or specific for a cell surface marker.","The targeted cells can be separated and/or sorted from cells lacking the marker by any of a variety of techniques.","In general, such techniques make use of a tag component associated with the specific binding molecule.","For example, the tag component can be a fluorescent label, a paramagnetic bead, or column matrix.","Fluorescent labels are useful for cell separation based on the well-established technique of fluorescence activated cell sorting (FACS).","Likewise, paramagnetic beads and the like and affinity columns derivatized with appropriate specific binding molecules can be used to separate cells.","Examples of fluorescent labels that can be used as tag components include fluorescein isothiocyanate (FITC), 5,6-carboxymethyl fluorescein, Texas red, nitrobenz-2-oxa-1,3-diazol-4-yl (NBD), coumarin, dansyl chloride, rhodamine, amino-methyl coumarin (AMCA), Eosin, Erythrosin, BODIPY®, Cascade Blue®, Oregon Green®, pyrene, lissamine, xanthenes, acridines, oxazines, phycoerythrin, macrocyclic chelates of lanthanide ions such as quantum dye™, fluorescent energy transfer dyes, such as thiazole orange-ethidium heterodimer, and the cyanine dyes Cy3, Cy3.5, Cy5, Cy5.5 and Cy7.Examples of other specific fluorescent labels that can be used as tag components include 3-Hydroxypyrene 5,8,10-Tri Sulfonic acid, 5-Hydroxy Tryptamine (5-HT), Acid Fuchsin, Alizarin Complexon, Alizarin Red, Allophycocyanin, Aminocoumarin, Anthroyl Stearate, Astrazon Brilliant Red 4G, Astrazon Orange R, Astrazon Red 6B, Astrazon Yellow 7 GLL, Atabrine, Auramine, Aurophosphine, Aurophosphine G, BAO 9 (Bisaminophenyloxadiazole), BCECF, Berberine Sulphate, Bisbenzamide, Blancophor FFG Solution, Blancophor SV, Bodipy F1, Brilliant Sulphoflavin FF, Calcien Blue, Calcium Green, Calcofluor RW Solution, Calcofluor White, Calcophor White ABT Solution, Calcophor White Standard Solution, Carbostyryl, Cascade Yellow, Catecholamine, Chinacrine, Coriphosphine O, Coumarin-Phalloidin, CY3.18, CY5.18, CY7, Dans (1-Dimethyl Amino Naphaline 5 Sulphonic Acid), Dansa (Diamino Naphtyl Sulphonic Acid), Dansyl NH—CH3, Diamino Phenyl Oxydiazole (DAO), Dimethylamino-5-Sulphonic acid, Dipyrrometheneboron Difluoride, Diphenyl Brilliant Flavine 7GFF, Dopamine, Erythrosin ITC, Euchrysin, FIF (Formaldehyde Induced Fluorescence), Flazo Orange, Fluo 3, Fluorescamine, Fura-2, Genacryl Brilliant Red B, Genacryl Brilliant Yellow 10GF, Genacryl Pink 3G, Genacryl Yellow 5GF, Gloxalic Acid, Granular Blue, Haematoporphyrin, Indo-1, Intrawhite Cf Liquid, Leucophor PAF, Leucophor SF, Leucophor WS, Lissamine Rhodamine B200 (RD200), Lucifer Yellow CH, Lucifer Yellow VS, Magdala Red, Marina Blue, Maxilon Brilliant Flavin 10 GFF, Maxilon Brilliant Flavin 8 GFF, MPS (Methyl Green Pyronine Stilbene), Mithramycin, NBD Amine, Nitrobenzoxadidole, Noradrenaline, Nuclear Fast Red, Nuclear Yellow, Nylosan Brilliant Flavin E8G, Oxadiazole, Pacific Blue, Pararosaniline (Feulgen), Phorwite AR Solution, Phorwite BKL, Phorwite Rev, Phorwite RPA, Phosphine 3R, Phthalocyanine, Phycoerythrin R, Polyazaindacene Pontochrome Blue Black, Porphyrin, Primuline, Procion Yellow, Pyronine, Pyronine B, Pyrozal Brilliant Flavin 7GF, Quinacrine Mustard, Rhodamine 123, Rhodamine 5 GLD, Rhodamine 6G, Rhodamine B, Rhodamine B 200, Rhodamine B Extra, Rhodamine BB, Rhodamine BG, Rhodamine WT, Serotonin, Sevron Brilliant Red 2B, Sevron Brilliant Red 4G, Sevron Brilliant Red B, Sevron Orange, Sevron Yellow L, SITS (Primuline), SITS (Stilbene Isothiosulphonic acid), Stilbene, Snarf 1, sulpho Rhodamine B Can C, Sulpho Rhodamine G Extra, Tetracycline, Thiazine Red R, Thioflavin S, Thioflavin TCN, Thioflavin 5, Thiolyte, Thiozol Orange, Tinopol CBS, True Blue, Ultralite, Uranine B, Uvitex SFC, Xylene Orange, and XRITC.","D. Antigens The disclosed methods make use of antigens to affect the immune response to those antigens.","For example, both antigens that cause desirable immune responses, such as antigens derived from pathogens (to be used to induce a protective immune response to the pathogen), and antigens that cause undesirable immune responses, such as antigens involved in auto-immune diseases, allergic reactions and graft rejection.","Any antigen of interest can be used.","Useful antigens include those involved in an immune response and/or disease or condition.","Particularly useful antigens are those that can cause an undesirable immune response.","Examples include antigens involved in auto-immune disease, allograft rejection, allergic reactions, allergic immune responses.","Useful antigens include those involved in auto-immune disease (referred to herein as auto-immune antigens), such as antigens involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.","Useful antigens also include those involved in allograft rejection, such as graft rejection of hematopoietic stem cell transplants.","Particularly useful antigens include MHC, QA, HIV gag, pol, and env (DNA or protein), rheumatoid factor, ICA 89, peripherin, carboxypeptidase H, glutamic acid decarboxylase (GAD), AHNAK, mylein basic protein, retinal S antigen, galactomannoprotein, neuraminidase, influenza matrix protein M1, CMV env proteins pp60, ds DNA, thyroglobulin, insulin, pancreatic islet beta cell antigens, MART-1 (melanoma protein), tyrosinase-related protein 2 (TRP2), melanoma cell lysate, MAGE-A3, allogeneic cell lysate, HLA antigens (protein or DNA), CD31 (protein or DNA), aspergillus chitin, pancreatic carcinoma cell line Panc-1 lysate, ErbB-2/neu, human epithelial cell mucin (Muc-1), Ha-ras oncogene product, p53, carcino-embryonic antigen (CEA), raf oncogene product, gp100/pmel17, GD2, GD3, GM2, TF, sTn, MAGE-1, MAGE-3, BAGE, GAGE, tyrosinase, gp75, Melan-A/Mart-1, gp100, HER2/neu, EBV-LMP 1 & 2, HPV-F4, 6, 7, prostate-specific antigen (PSA), prostate specific membrane antigen (PSMA), HPV-16, MUM, alpha-fetoprotein (AFP), CO17-1A, GA733, gp72, p53, ras oncogene product, HPV E7, Wilm's tumor antigen-1, telomerase, melanoma gangliosides, malignant B cell antigen receptor, malignant B cell immunoglobulin idiotype, variable region of an immunoglobulin, hypervariable region or complementarity determining region (CDR) of a variable region of an immunoglobulin, malignant T cell receptor (TCR), variable region of a TCR, and hypervariable region of a TCR.","For induction of an immune response against cancer or tumor cells, the antigens can be tumor antigens.","Tumor antigens for use in the disclosed method can be any tumor antigen now known or later identified as a tumor antigen.","The appropriate tumor antigen used depends on the tumor type being treated.","For example, the tumor antigen can be, but is not limited to human epithelial cell mucin (Muc-1; a 20 amino acid core repeat for Muc-1 glycoprotein, present on breast cancer cells and pancreatic cancer cells), the Ha-ras oncogene product, p53, carcino-embryonic antigen (CEA), the raf oncogene product, gp100/pmel17, GD2, GD3, GM2, TF, sTn, tyrosinase-related protein 2 (TRP2), melanoma cell lysate, MAGE-A3, MAGE-1, MAGE-3, BAGE, GAGE, tyrosinase, gp75, Melan-A/Mart-1, gp100, HER2/neu, ErbB-2/neu, pancreatic carcinoma cell line Panc-1 lysate, EBV-LMP 1 & 2, HPV-F4, 6, 7, prostate-specific antigen (PSA), prostate specific membrane antigen (PSMA), HPV-16, MUM, alpha-fetoprotein (AFP), CO17-1A, GA733, gp72, p53, the ras oncogene product, HPV E7, Wilm's tumor antigen-1, telomerase, and melanoma gangliosides, as well as any other tumor antigens now known or identified in the future.","Tumor antigens can be obtained following known procedures or are commercially available.","Additionally, the tumor antigen of the present invention can be an antibody which can be produced by a B cell tumor (e.g., B cell lymphoma; B cell leukemia; myeloma) or the tumor antigen can be a fragment of such an antibody, which contains an epitope of the idiotype of the antibody.","The epitope fragment can comprise as few as nine amino acids.","For example, the tumor antigen of this invention can be a malignant B cell antigen receptor, a malignant B cell immunoglobulin idiotype, a variable region of an immunoglobulin, a hypervariable region or complementarity determining region (CDR) of a variable region of an immunoglobulin, a malignant T cell receptor (TCR), a variable region of a TCR and/or a hypervariable region of a TCR.","Where the object of treatment is graft rejection, graft versus host disease, or auto-immune disease, the antigens can be prepared using cells from the source of the antigen(s) involved.","For example, cells from the source of transplant cells can be used to obtain relevant antigens.","As will be appreciated, the source cells contain antigens that are involved in graft rejection.","Cells from the individual to be treated can be used to obtain relevant antigens in the case of auto-immune diseases.","In general, whole cell preparations can be used.","The cells can irradiated to kill the cells and the resulting treated cell mixture can be used as the antigens for incubation with dendritic cells of loading into liposomes targeted to dendritic cells.","As a specific example, allo-antigen can be prepared by irradiating peripheral blood mononuclear cells from a third party donor to 25 Gy.","Use of antigens prepared in this way is described in the Example.","Useful antigens for altering immune responses in the context of transplants include MHC and QA antigens.","Useful antigens for altering immune responses to influenza include neuraminidase, and influenza matrix protein M1.Useful antigens for altering immune responses to CMV include env proteins pp60.Useful antigens for altering immune responses in lupus include ds DNA and AHNAK.","Useful antigens for altering immune responses in thyroditis include thyroglobulin.","Useful antigens for altering immune responses in diabetes include insulin and pancreatic islet beta cell antigens.","Useful antigens for altering immune responses to allografts include allogeneic cell lysate, HLA antigens (protein or DNA), and CD31 (protein or DNA).","Useful antigens for altering auto-immune responses include rheumatoid factor, ICA 89, peripherin, carboxypeptidase H, and glutamic acid decarboxylase (GAD).","Useful antigens for altering immune responses to fungal infections include aspergillus chitin and galactomannoprotein.","Useful antigens for altering immune responses to pancreatic cancer include pancreatic carcinoma cell line Panc-1 lysate.","Useful antigens for altering immune responses to prostate cancer include prostate specific antigen.","Useful antigens for altering immune responses to colon cancer include carcioembryonic antigen.","Useful antigens for altering immune responses to breast, renal, and/or lung cancer include ErbB-2/neu.","Useful antigens for altering immune responses to HIV include HIV gag, pol, and env (DNA or protein).","E. Liposomes The disclosed liposomes comprise liposomes comprising one or more antigens and one or more surface-bound molecules.","The antigens are antigens of interest, such as antigens involved in undesirable immune responses.","Liposomes comprising antigens are referred to herein as antigen-loaded liposomes.","The surface-bound molecules are molecules that target the liposomes to dendritic cells.","Liposomes containing surface-bound molecules are referred to herein as targeted liposomes.","For convenience, the term liposome is used herein to refer to the base liposome, as well as to antigen-loaded and targeted liposomes.","Generally, the context of the term indicates what is meant.","Surface-bound molecules generally comprise a component that can bind to and/or interact with targeted cells.","In the context of the disclosed method, the targeted cells are dendritic cells.","Useful components that can bind to and/or interact with targeted cells are specific binding molecules.","In the context of the disclosed method, the specific binding molecule is generally interacts with and/or is specific for a cell surface marker.","Particularly useful targets for surface-bound molecules are CD11c, BDCA-1, or both, for targeting mature type 1 dendritic cells, and CD123, BDCA-2, BDCA-4, or a combination, for targeting type 2 dendritic cells.","Liposomes are artificial structures primarily composed of phospholipid bilayers.","Cholesterol and fatty acids may also be included in the bilayer construction.","Liposomes may be loaded with compounds of interest, such as the disclosed antigens, and coated on the outer surface with surface-bound molecules, such as specific binding molecules.","Liposomes, preferably unilamellar vesicles, can be made using established procedures that result in the loading of the interior compartment with antigens.","The liposomes can also be associated with a surface-bound molecules, such as a specific binding molecule.","The association may be direct or indirect.","An example of a direct association is a liposome containing covalently bound antibodies on the surface of the phospholipid bilayer.","An example of indirect association is a liposome containing covalently bound anti-antibody antibodies that can bind targeted antibodies.","Liposome-forming compounds are generally well known as are the methods of their preparation.","For example, any number of phospholipids or lipid compounds may be used to form the vesicle walls.","Representative of such wall forming compounds are; phosphatidylcholine (hereinafter referred to as “PC”), both naturally occurring and synthetically prepared, phosphatidic acid (hereinafter referred to as “PA”), lysophosphatidylcholine, phosphatidylserine (hereinafter referred to as “PS”), phosphatidylethanolamine (hereinafter referred to as “PE”), sphingolipids, phosphatidyglycerol (hereinafter referred to as “PG”), spingomyelin, cardiolipin, glycolipids, gangliosides, cerebrosides and the like used either singularly or intermixed such as in soybean phospholipids.","In addition, other lipids such as steroids, cholesterol, aliphatic amines such as long chain aliphatic amines and carboxylic acids, long chain sulfates and phosphates, dicetyl phosphate, butylated hydroxytoluene, tocophenol, retinol, and isoprenoid compounds may be intermixed with the phospholipid components to confer certain desired and known properties on the formed vesicles.","In addition, synthetic phospholipids containing either altered aliphatic portions such as hydroxyl groups, branched carbon chains, cycloderivatives, aromatic derivatives, ethers, amides, polyunsaturated derivatives, halogenated derivatives or altered hydrophillic portions containing carbohydrate, glycol, phosphate, phosphonate, quaternary amine, sulfate, sulfonate, carboxy, amine, sulfhydryl, imidazole groups and combinations of such groups can be either substituted or intermixed with the above mentioned phospholipids and used in the disclosed method.","It will be appreciated from the above that the chemical composition of the lipid component of the vesicles prepared by the method of the invention may be varied greatly without appreciable diminution of percentage capture although the size of the vesicle may be affected by the lipid composition.","A convenient mixture we have used extensively and which is representative of lipid mixtures advantageously used in the method of the invention is composed of PS and PC, or PG and PC as identified above (advantageously at a 1:4 molar ratio in each instance).","The PC, PG, PA and PE, may be derived from purified egg yolk.","Saturated synthetic PC and PG, such as dipalmitoyl may also be used.","Other amphipathic lipids that may be used, advantageously also at 1:4 molar ratios with PC, are gangliosides, globosides, fatty acids, stearylamine, long chain alcohols, and the like.","The production and use of liposomes, including description of liposomes having useful properties and specialized components, are known.","For example, various liposomes, their components and their method of production and use are described in U.S. Pat.","No.","6,296,870, U.S. Pat.","No.","6,241,999, U.S. Pat.","No.","6,214,388, U.S. Pat.","No.","5,354,853, U.S. Pat.","No.","4,900,556, U.S. Pat.","No.","4,708,861, U.S. Pat.","No.","4,235,871, and U.S. Pat.","No.","4,224,179, all of which are incorporated herein by reference for their description of he above subject matter.","The class of liposomes described in U.S. Pat.","No.","6,214,388, which are designed to maximize internalization of liposomes by cells having cell surface markers of interest, are particularly useful in the disclosed method.","1.Modifying Liposomes Disclosed is a method of modifying liposomes comprising loading liposomes with one or more antigens.","The liposomes can also be modified by adding surface-bound molecules to the surface of the liposomes.","The surface-bound molecules can target the liposomes to cells, such as dendritic cells.","Techniques for loading liposomes and for producing liposome having surface-bound molecules are known.","For example, liposomes can be loaded with antigens by forming the liposomes in the presence of the antigens (see, for example, liposome patents described above).","Liposomes having surface-bound molecules can be produced, for example, by using lipids to which the surface-bound molecules are couple or otherwise attached.","It is useful to couple surface-bound molecules at or near a polar or charged portion of the lipids.","This should allow the surface-bound molecules to be exposed on the surface of the liposomes.","F. Specific Binding Molecules A specific binding molecule is a molecule that interacts specifically with a particular molecule or moiety.","In the context of the disclosed method, the molecule or moiety that interacts specifically with a specific binding molecule is generally a cell surface marker.","Antibodies, either member of a receptor/ligand pair, synthetic polyamides (Dervan and Burli, Sequence-specific DNA recognition by polyamides.","Curr Opin Chem Biol, 3(6):688-93 (1999); Wemmer and Dervan, Targeting the minor groove of DNA.","Curr Opin Struct Biol, 7(3):355-61 (1997)), and other molecules with specific binding affinities are examples of specific binding molecules.","Specific binding molecules are generally used to isolate useful dendritic cells and/or to target liposomes to dendritic cells as described elsewhere herein.","A specific binding molecule that interacts specifically with a particular analyte is said to be specific for that analyte.","For example, where the specific binding molecule is an antibody that associates with a particular antigen, the specific binding molecule is said to be specific for that antigen.","The antigen is the analyte.","A reporter molecule containing the specific binding molecule can also be referred to as being specific for a particular analyte.","Specific binding molecules preferably are antibodies, ligands, binding proteins, receptor proteins, haptens, aptamers, carbohydrates, synthetic polyamides, peptide nucleic acids, or oligonucleotides.","Preferred binding proteins are DNA binding proteins.","Preferred DNA binding proteins are zinc finger motifs, leucine zipper motifs, helix-turn-helix motifs.","These motifs can be combined in the same specific binding molecule.","1.Antibodies Antibodies useful as specific binding molecules in the disclosed method are generally specific for cell surface markers.","Numerous antibodies specific for various cell surface markers are known, and many are commercially available.","Such antibodies can be used in the disclosed method and/or to produce components used in the disclosed method (such as for use as specific binding molecules).","Useful antibodies can also be produced for use in the disclosed method.","Techniques for antibody production are known, some of which are described below.","Such techniques can be used to produce antibodies for use in the disclosed method.","The term “antibodies” is used herein in a broad sense and includes both polyclonal and monoclonal antibodies.","In addition to intact immunoglobulin molecules, also included in the term “antibodies” are fragments or polymers of those immunoglobulin molecules, and human or humanized versions of immunoglobulin molecules or fragments thereof, as long as they are chosen for their ability to interact with cell surface markers (or other antibody target), as described herein.","The antibodies can be tested for their desired activity using the in vitro assays described herein, or by analogous methods.","The term “monoclonal antibody” as used herein refers to an antibody obtained from a substantially homogeneous population of antibodies, that is, the individual antibodies within the population are identical except for possible naturally occurring mutations that may be present in a small subset of the antibody molecules.","The monoclonal antibodies herein specifically include “chimeric” antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, as long as they exhibit the desired antagonistic activity (See, U.S. Pat.","No.","4,816,567 and Morrison et al., Proc.","Natl.","Acad.","Sci.","USA, 81:6851-6855 (1984)).","Monoclonal antibodies can be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature, 256:495 (1975).","In a hybridoma method, a mouse or other appropriate host animal is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent.","Alternatively, the lymphocytes may be immunized in vitro, e.g., using the HIV Env-CD4-co-receptor complexes described herein.","The monoclonal antibodies may also be made by recombinant DNA methods, such as those described in U.S. Pat.","No.","4,816,567 (Cabilly et al.).","DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (for example, by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies).","Libraries of antibodies or active antibody fragments can also be generated and screened using phage display techniques, for example, as described in U.S. Pat.","No.","5,804,440 to Burton et al.","and U.S. Pat.","No.","6,096,441 to Barbas et al.","In vitro methods are also suitable for preparing monovalent antibodies.","Digestion of antibodies to produce fragments thereof, particularly, Fab fragments, can be accomplished using routine techniques known in the art.","For instance, digestion can be performed using papain.","Examples of papain digestion are described in WO 94/29348 published Dec. 22, 1994 and U.S. Pat.","No.","4,342,566.Papain digestion of antibodies typically produces two identical antigen binding fragments, called Fab fragments, each with a single antigen binding site, and a residual Fc fragment.","Pepsin treatment yields a fragment that has two antigen combining sites and is still capable of cross-linking antigen.","The fragments, whether attached to other sequences or not, can also include insertions, deletions, substitutions, or other selected modifications of particular regions or specific amino acids residues, provided the activity of the antibody or antibody fragment is not significantly altered or impaired compared to the non-modified antibody or antibody fragment.","These modifications can provide for some additional property, such as to remove/add amino acids capable of disulfide bonding, to increase its bio-longevity, to alter its secretory characteristics, etc.","In any case, the antibody or antibody fragment must possess a bioactive property, such as specific binding to its cognate antigen.","Functional or active regions of the antibody or antibody fragment may be identified by mutagenesis of a specific region of the protein, followed by expression and testing of the expressed polypeptide.","Such methods are readily apparent to a skilled practitioner in the art and can include site-specific mutagenesis of the nucleic acid encoding the antibody or antibody fragment.","(Zoller, M. J. Curr.","Opin.","Biotechnol.","3:348-354, 1992).","As used herein, the term “antibody” or “antibodies” can also refer to a human antibody and/or a humanized antibody.","G. Tag Components Tag components are molecules that can be associated with specific bindingmolecules and used to separate or sort cells.","Useful tag components include fluorescent molecules and sortable components.","Fluorescent molecules are particularly useful for fluorescence activated cell sorting (FACS).","Sortable components include molecules and compositions that can be selectively bound or that can selectively interact with other molecules and compounds.","Examples include beads or particles, such as paramagnetic bead, that can be separated based on a property of the bead.","Thus, for example, cells associated with paramagnetic bead or particle can be selected or separated by magnetic sorting.","Specific binding molecules are a useful form of tag component.","Cells associated with such a specific binding molecule can be separated by associating the specific binding molecule with its binding partner (that is the molecule or moiety with which the specific binding molecule interacts specifically).","Thus, for example, cells with which a specific binding molecule (as tag component) is associated can be selected or separated by passing the cells over a column derivatized with the binding partner of the specific binding molecule.","H. Incubation of Dendritic Cells With Antigens Some forms of the disclosed methods involve or make use of dendritic cells that have been incubated with or exposed to one or more antigens.","In particular, the disclosed method can use type 2 dendritic cells or CD11c−, CD123bright, BDCA-2+ dendritic cells that have been incubated with one or more antigens.","Administration of such dendritic cells to an individual can induce immunological tolerance to the antigens.","Alternatively, the disclosed method can use immature type 1 dendritic cells or CD11c−, CD123dim, BDCA-2− dendritic cells that have been incubated with one or more antigens.","Administration of such type 1 dendritic cells to an individual can enhance immune responses to, for example, tumor antigens and antigens associated with infectious diseases.","Dendritic cells are generally incubated with antigens ex vivo.","In this context, ex vivo refers to treatment of the cells outside of the body from which they are derived or isolated.","If ex vivo methods are employed, the cells can be maintained outside the body according to standard protocols well known in the art.","The antigens can be incubated with the cells under any suitable conditions.","Useful conditions include conditions useful for maintenance of the cells outside the body.","As a specific example, preparations of antigens from irradiated cells can be incubated with dendritic cells.","For example, 20,000 irradiated cells can be added to isolated dendritic cells and the cells incubated for 24 and 72 hours in a 5% CO2 atmosphere at 37° C. TNF can then be added (at, for example, 10 ng/ml) to mature and activate dendritic cell progenitors.","Use of these incubation conditions is described in the Example.","I.","Administration of Dendritic Cells Dendritic cells exposed to antigen can be administered to individuals and animals using any suitable technique.","In general, The treated cells can be infused per standard methods for the cell type.","Standard methods are known for infusion of dendritic cells into a subject.","In general, the disclosed dendritic cells can be administered by infusion, such as by intravenous infusion.","A useful method of intraveneous infusion is to insert a indwelling catherter into a vein and attach to the catheter a flexible tubing, usually composed of plastic, and attach to the tubing a reservoir containing the substance to be infused.","The reservoir may be a bag or syringe containing a liquid solution.","The solution to be infused is transferred from the reservoir bag through the flexible tubing and indwelling catherter into the venous circulation of the recipient.","The force to affect the transfer of the solution into the venous circulation may be gravity or compression of the bag or syringe.","However, the dendritic cells may also be administered orally, parenterally (for example, intravenously), by intramuscular injection, by intraperitoneal injection, transdermally, sub-cutaneously, topically or the like.","As used herein, “intravenous infusion” means delivery of the cells into a vein of the individual.","Administration of the dendritic cells by inhalant can be through the nose or mouth via delivery by a spraying or droplet mechanism.","J.","Administration of Liposomes Liposomes can be administered to individuals and animals using any suitable technique.","In general, the disclosed liposomes can be administered by infusion, such as by intravenous infusion.","This is useful because the targeted dendritic cells are found in the blood.","However, the liposomes may be administered orally, parenterally (for example, intravenously), by intramuscular injection, by intraperitoneal injection, transdermally, sub-cutaneously, topically or the like.","As used herein, “intravenous infusion” means delivery of the liposomes into a vein of the individual.","Administration of the liposomes by inhalant can be through the nose or mouth via delivery by a spraying or droplet mechanism.","Delivery can also be directly to any area of the respiratory system (for example, lungs) via intubation.","The exact amount of the liposomes required will vary from subject to subject, depending on the species, age, weight and general condition of the subject, the mode of administration and the like.","Thus, it is not possible to specify an exact amount for every liposome.","However, an appropriate amount can be determined by one of ordinary skill in the art using only routine experimentation given the teachings herein.","K. Immune Responses An immune response refers to one or more effects produced by the immune system in reaction to one or more antigens.","This includes, for example, immune system reactions to combinations of antigens; combinations of one or more antigens with other, non-antigenic components; and antigens in or on cells, viruses, liposomes or other compositions.","As used herein, immune response refers to the anti-antigen function of the immune system and not to effects on and changes in the immune system that reduce the anti-antigen function of the immune system.","For example, stimulation of neutralizing antibodies in response to exposure of an immune system to an antigen is an immune response, while induction of immunological tolerance is not an immune response as that term is used herein.","It should be understood that this is the case even though immunological tolerance represents an effect produced by the immune system in reaction to one or more antigens.","Adaptive immune response can largely be differentiated into two components humoral and cell-mediated.","This division is based on whether the immune response is dominated by antibodies produced by plasma cells (a B cell lineage cell) or cytotoxic T cells.","However, rarely are immune responses strictly the domain of either one of the components.","Predominantly, the immune response is a combination of the two arms of the immune response having both T cell and B cell components.","The dominance of either component is largely the result of regulatory CD4+ T cells, and the interaction of these cells with antigen in the context of dendritic cells.","CD4 T cells can produce a myriad of functions such as cytokine release and direct cytotoxicity.","It is these functions that determines whether the cell-mediated or humoral responses will dominate.","Typically, CD4 T cells that stimulate a cell-mediate response are referred to as inflammatory CD4 T cells or TH1 cells.","A TH1 response is characterized by IFN-γ, TNF, and IL 12 production from CD8 and CD4 T cells, IgG2a antibody production, and the generation of cytotoxic T cells.","CD4 cells that induce a more humoral response are termed TH2.TH2 responses result in IL-4, IL5, and IL10 secretion and IgG1, IgA and IgE antibody production.","The determination as to whether a CD4 cell will become a TH1 or TH2 cell is largely a factor of the environment that surround the cell at the time of activation.","The environmental factors that control this are cytokines and the type of “non-professional” antigen-presenting cells or “professional” antigen-presenting cells (dendritic cells) that presents the antigen.","Secreted IL-12 and IFN-γ in the presence of an uncommitted T cell can promote a TH1 response.","Similarly, secreted IL-4 in the presence of an uncommitted T cell can promote a TH2 response.","Additionally, these cytokines have the effect of suppressing the opposite response.","The environment associated with the determination of the commitment to a TH1 or TH2 pathway also involves the cells that present antigen to CD4 T cells.","CD4 T cells recognize peptide antigen in the context of MHC class II molecules presented by antigen presenting cells.","These cells may be macrophage, mast cells, dendritic cells, or B cells.","Depending on the cell type that presents the antigen the CD4 cell may be committed to either pathway.","CD4 T cells activated by macrophage or type 1 dendritic cells are directed to induce a cell-mediated immune response.","CD4 T cells activated by type 2 dendritic cells and mast cells are directed to induce a TH2 immune response.","1.Reduction of Immune Responses Some embodiments of the disclosed methods result in reduction of an immune response.","As used herein, reduction of an immune response refers to a lessening of one or more effects produced by the immune system in reaction to an antigen.","Such reduction includes both reduction of an actual immune response or a reduction in an immune response that would otherwise have been expected in the absence of the reducing treatment.","Reduction can include, but does not require, lessening of the immune response to an undetectable level.","As used herein, suppression of an immune response refers to a lessening of one or more effects produced by the immune system in reaction to an antigen.","Such suppression includes both lessening of an actual immune response or a lessening of an immune response that would otherwise have been expected in the absence of the suppressing treatment.","Suppression can include, but does not require, lessening of the immune response to an undetectable level.","2.Enhancement of Immune Responses Some embodiments of the disclosed methods result in enhancement of an immune response.","As used herein, enhancement of an immune response refers to an increase of one or more effects produced by the immune system in reaction to an antigen.","Such enhancement includes both enhancement of an actual immune response or an enhancement in an immune response that would otherwise not have been expected in the absence of the enhancing treatment.","As used herein, generation of an immune response refers to the creation of one or more effects produced by the immune system (to a detectable level) in reaction to an antigen.","Such generation includes creation of an immune response that would otherwise not have been expected in the absence of the generating treatment.","L. Immunological Tolerance Some forms of the disclosed methods result in immunological tolerance to one or more antigens.","As used herein, immunological tolerance refers to a physiological state in which the immune system does not react destructively against antigens.","In effect, immunological tolerance is a lack of an immune response to an antigen.","Mechanistically, immunological tolerance generally is the result of either deletion of reactive cells during development, such as “central tolerance” that occurs during deletion of auto-reactive T-cells during intrathymic development, or peripheral tolerance, that may occur during encounter of immune cells with antigen at peripheral lymphoid tissues in a manner that generates an anergis or unresponsive state.","Immunological tolerance generally involves interruption of one or more steps involved in generation of an immune response.","For example, individuals generally have immunological tolerance to self-antigens.","This tolerance is a result of negative selection that takes place during T and B cell development in the thymus and bone marrow respectively.","For T cells, this process involves the clonal deletion of T cells that recognize self-antigens.","The deletion takes place in the thymus.","Here CD4+CD8+ T cells are transitioning to either CD4−CD8+ or CD4+CD8− T cells.","At this point in their development, the T cells are positively selected based on exposure and the subsequent ability to bind self-MHC.","T cells that do not respond to the stimuli undergo activation induced cell death (AICD) and are deleted from the T cell repertoire, as being unable to recognize self-MHC would prevent the recognition of antigen since a TCR will only recognize antigen in the context of MHC.","Those T cells that recognize self-MHC (that is, bind above a certain threshold) finish the differentiation to CD4−CD8+ or CD4+CD8− T cells.","The surviving T cells are then negatively selected if they become activated by self-antigen being presented on self-MHC.","This negative selection insures that the immune system won't respond to self and the responding T cells are deleted.","Interestingly, foreign antigen present in the organism at the time of negative selection will also be seen as self.","An example of this is the creation of Lymphocytic choriomeningitis virus (LCMV) carrier mice via neonatal infection.","Adult mice infected with LCMV have a profound T cell response which can lead to meningitis due to immune pathology.","Though a persistent infection can ensue, mice not dying via immune pathology will clear the infection.","However, neonatal mice infected with LCMV will not generate an immune response and will become lifelong carriers of the virus showing no ill effects of infection or immune pathology.","Similarly, mice receiving a non-lethal dose of irradiation will lose all T cells.","Exposure to a foreign antigen prior to the reconstitution of the T cell population will result in animals that are tolerant to the foreign antigen.","Though the mechanism of central tolerance through clonal deletion insures that the majority of T cells do not recognize self, there are tissue-specific antigens in the periphery that will not be presented to developing T cells in the thymus.","Therefore, it is still possible to have T cells that react to self.","However, the activation of a T cell is a two step process (1) the T cell must recognize antigen in the context of self-MHC and (2) the T cell must receive co-stimulatory signals in the form of CTLA-4-CD80/CD86, 41BB-41BBL, CD40-CD40L or CD28-B7.1/B7.2 interactions.","Without both signals, a T cell will not become activated.","In the situation of peripheral antigens, T cells specific for these self-antigens do not receive a co-stimulatory signal and therefore become anergic.","In contrast to T cells, B cells do not recognize antigen in the context of MHC and therefore do not require a positive selection step.","However, immature B cells do undergo negative selection within the bone marrow.","Here, B cells with immunoglobulin (BCR) that recognizes self-antigens are clonally deleted.","This was shown experimentally in mice transgenic for transmembrane bound hen-egg lysozyme (HEL) and transgenic for B cells specific for HEL.","In these experiments, immature B cells specific for HEL can be detected, but no HEL specific mature B cells exist as they are deleted from the repertoire.","Despite the deletion of self-specific cells in the bone marrow, there is still potential for mature B cells to exist that recognize soluble self-antigen or antigen only expressed in certain peripheral tissue.","In the latter situation, B cells that recognize antigen expressed in peripheral tissue are deleted.","However, B cells that recognize soluble antigen are not deleted.","Instead these B cells internalize their antigen receptor and lose functionality.","This loss of function is called anergy and results in a lack of response to an antigen despite the presence of specific cells.","M. Pharmaceutically Acceptable Carriers The disclosed cells and liposomes can be used therapeutically in combination with a pharmaceutically acceptable carrier.","Pharmaceutical carriers are known to those skilled in the art.","These most typically would be standard carriers for administration of drugs to humans, including solutions such as sterile water, saline, and buffered solutions at physiological pH.","Particularly useful pharmaceutical carriers are physiological solutions.","In the case of the disclosed cells, media and solutions in which the cells are store, grown, maintained or.","incubated are particularly useful as pharmaceutical carriers.","Pharmaceutical compositions may include carriers, thickeners, diluents, buffers, preservatives, surface active agents and the like in addition to the cells or liposomes.","Pharmaceutical compositions may also include one or more active ingredients such as antimicrobial agents, anti-inflammatory agents, anesthetics, and the like.","It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope or spirit of the invention.","Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein.","It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being indicated by the following claims.","EXAMPLE A.","The Effect of DC2p Depletion on the Ability of the Cells in an HPC Product to Generate an Allo-Immune Response.","The mononuclear cell fraction of the blood from a normal donor was collected by apheresis.","Naïve CD45RA+ 62L+ T-cells were purified by FACS and frozen.","The HPC product was depleted of DC2p using the BDCA2 antibody and the MACS immuno-affinity columns or purified DC1p and FACS isolated DC2p populations.","The ability of the DC2p depleted HPC cells and the purified DCp2 and DC1p populations to generate cellular immune responses to allo-antigen was determined by adding allo-antigen to the DCp cells, maturing the DCp by the addition of IL3, GM-CSF, and TNF, then adding the previously collected naïve T-cells and measuring their proliferation in response to the allo-antigen (FIGS.","1 and 2).","The proliferative response of autologous T-cells to allo-antigen presented by purified DC2p was inhibited, while T-cells showed marked proliferation in response to allo-antigen presented by purified DC1p.","Of note, T-cells had a strong proliferative response to the HPC graft that was depleted of the inhibitory DC2p population (stars, FIG.","2).","Procedural Details.","Peripheral blood mononuclear cells were obtained form a normal donor by apheresis.","Naïve CD3+, CD45RA+, CD62L+ naïve T-cells were isolated following staining with fluorescently labeled anti-CD3, anti-CD45RA, and anti 62L and high speed fluorescent activated cell sorting.","DC1 (CD11c+, CD123dim, lineage-) and DC2 (CD11c−, CD123+, lineage-) populations were enriched from the apheresis product using immuno-affinity magnetic purification using antibodies to DC1 (BDCA-1) and DC2 (BDCA-2) then purified to >95% homogeneity by high speed cell sorting after staining with monoclonal antibodies to CD123, CD11c and a cocktail of lineage markers (CD3, CD14, CD19, and CDF56).","Defined numbers of mononuclear cells (start fraction), purified DC1, purified DC2, or mononuclear cells depleted of either DC1 or DC2 by immuno-magnetic affinity column chromatography were added to tissue culture wells in a 96 well plate and cultured with 50 ng/ml GM-CSF and 50 ng/ml IL3 (start, DC2, and DC1 depleted) or 50 ng/ml GM-CSF and 50 ng/ml IL4 (DC1, DC2 depleted).","Allo-antigen was prepared by irradiating peripheral blood mononuclear cells from a third party donor to 25 Gy then adding 20,000 cells to each well containing stimulator cells.","After 24 and 72 hours of culture in a 5% CO2 atmosphere at 37° C., TNF was added at 10 ng/ml to mature and activate DC progenitors.","50,000 autologous naïve T-cells were added to each well and tritiated thymidine incorporation was measured after an additional 96 hours of culture.","FIGS.","1 and 2 show the results.","It is understood that the disclosed invention is not limited to the particular methodology, protocols, and reagents described as these may vary.","It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.","It must be noted that as used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural reference unless the context clearly dictates otherwise.","Thus, for example, reference to “a host cell” includes a plurality of such host cells, reference to “the antibody” is a reference to one or more antibodies and equivalents thereof known to those skilled in the art, and so forth.","Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of skill in the art to which the disclosed invention belongs.","Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, devices, and materials are as described.","Publications cited herein and the material for which they are cited are specifically incorporated by reference.","Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.","Throughout this application, various publications are referenced.","The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.","The references disclosed are also individually and specifically incorporated by reference herein for the material contained in them that is discussed in the sentence in which the reference is relied upon.","Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein.","Such equivalents are intended to be encompassed by the following claims."]],"string":"[\n [\n \"Regulating immune response using dendritic cells\",\n \"Disclosed within a method of altering immune responses using dendritic cells.\",\n \"One form of the method is a method of inducing immunological tolerance in an individual, where type 2 dendritic cells are administrated to an individual, and where the dendritic cells have been incubated with one or more antigens.\",\n \"Another form of the method involves altering an immune response, in which liposomes containing where liposomes containing one or more antigens are administrated to an individual, and where the liposomes are modified with surface-bound molecules that target the liposomes to type 2 dendritic cells.\",\n \"Another form of the method involves reducing immune responsiveness, where liposomes containing one or more antigens are administrated to an individual and where the liposomes are modified with the surface bound molecules that target the liposomes to type 1 dendritic cells or type 2 dendritic cells.\",\n \"Another form of the method is a method of enhancing immune responsiveness, where liposomes containing one or more antigens are administrated to an individual, and where the liposomes are modified with surface-bound molecules that target the liposomes to mature type 1 dendritic cells.\"\n ],\n [\n \"1-27.\",\n \"(canceled) 28.A method of altering an immune response comprising administering to the individual liposomes, wherein the liposomes comprise one or more antigens, wherein the liposomes are modified with surface-bound molecules that target the liposomes to type 2 dendritic cells or mature type 1 dendritic cells.\",\n \"29.The method of claim 28, wherein the antigens are carbohydrates.\",\n \"30.The method of claim 28, wherein the antigens are nucleic acids.\",\n \"31.The method of claim 28, wherein the antigens are peptides.\",\n \"32.The method of claim 28, wherein the antigens are lipids.\",\n \"33.The method of claim 28, wherein the antigens are a combination of two or more of the following: carbohydrates, nucleic acids, peptides, or lipids.\",\n \"34.The method of claim 28, wherein the antigens are MHC, QA, HIV gag, pol, and env (DNA or protein), rheumatoid factor, ICA 89, peripherin, carboxypeptidase H, glutamic acid decarboxylase (GAD), AHNAK, mylein basic protein, retinal S antigen, galactomannoprotein, neuraminidase, influenza matrix protein M1, CMV env proteins pp60, ds DNA, thyroglobulin, insulin, pancreatic islet beta cell antigens, MART-1 (melanoma protein), tyrosinase-related protein 2 (TRP2), melanoma cell lysate, MAGE-A3, allogeneic cell lysate, HLA antigens (protein or DNA), CD31 (protein or DNA), aspergillus chitin, pancreatic carcinoma cell line Panc-1 lysate, ErbB-2/neu, human epithelial cell mucin (Muc-1), Ha-ras oncogene product, p53, carcino-embryonic antigen (CEA), raf oncogene product, gp100/pmel17, GD2, GD3, GM2, TF, sTn, MAGE-1, MAGE-3, BAGE, GAGE, tyrosinase, gp75, Melan-A/Mart-1, gp100, HER2/neu, EBV-LMP 1 & 2, HPV-F4, 6, 7, prostate-specific antigen (PSA), prostate specific membrane antigen (PSMA), HPV-16, MUM, alpha-fetoprotein (AFP), CO17-IA, GA733, gp72, p53, ras oncogene product, HPV E7, Wilm's tumor antigen-1, telomerase, melanoma gangliosides, malignant B cell antigen receptor, malignant B cell immunoglobulin idiotype, variable region of an immunoglobulin, hypervariable region or complementarity determining region (CDR) of a variable region of an immunoglobulin, malignant T cell receptor (TCR), variable region of a TCR, hypervariable region of a TCR, or a combination.\",\n \"35.The method of claim 28, wherein the surface-bound molecule is a monoclonal antibody.\",\n \"36.The method of claim 28, wherein the surface-bound molecule is a peptide high-affinity ligand.\",\n \"37.The method of claim 28, wherein the surface-bound molecule is specific for CD11c+, BDCA-1, or both, wherein mature type 1 dendritic cells are targeted.\",\n \"38.The method of claim 37, wherein the liposomes enhance anti-tumor immune responses.\",\n \"39.The method of claim 37, wherein the liposomes enhance anti-cancer immune responses.\",\n \"40.The method of claim 28, wherein the surface-bound molecule is specific for CD123, BDCA-2, BDCA-4, or a combination, wherein type 2 dendritic cells are targeted.\",\n \"41.The method of claim 40, wherein the liposomes reduce an auto-immune response.\",\n \"42.The method of claim 41, wherein the auto-immune response is involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and systemic lupus erythematosis.\",\n \"43.The method of claim 40, wherein the liposomes reduce an allergic immune response.\",\n \"44.The method of claim 43, wherein the allergic immune response is involved in asthma, gluten allergy, and atopic dermatitis.\",\n \"45.The method of claim 40, wherein the liposomes reduce graft rejection immune response.\",\n \"46.The method of claim 45, wherein the individual is an allograft recipient.\",\n \"47.The method of claim 45, wherein the graft rejection immune response comprises complications associated with graft rejection.\",\n \"48.The method of claim 45, wherein the graft rejection immune response comprises graft rejection.\",\n \"49.The method of claim 40, wherein the individual is a hematopoietic stem cell recipient.\",\n \"50.The method of claim 40, wherein the liposomes reduce graft verses host immune response.\",\n \"51.The method of claim 40, wherein the antigens are auto-immune antigens.\",\n \"52.The method of claim 51, wherein the auto-immune antigens are involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, or systemic lupus erythematosis.\",\n \"53.The method of claim 28, wherein the antigens are antigens involved in allograft rejection, wherein the liposomes reduce the allograft rejection.\",\n \"54.The method of claim 28, wherein the antigens are antigens involved in graft rejection of hematopoietic stem cell transplants, wherein the liposomes reduce the graft rejection.\",\n \"55.The method of claim 28, wherein the antigens are antigens involved in graft verses host disease.\",\n \"56.A method of reducing an immune response comprising administering to the individual liposomes, wherein the liposomes comprise one or more antigens, wherein the liposomes are modified with surface-bound molecules that target the liposomes to type 2 dendritic cells.\",\n \"57.The method of claim 28, wherein the liposomes enhance an immune response, wherein the liposomes are targeted to immature type 1 dendritic cells.\",\n \"58.The method of claim 28, wherein the liposomes reduce an immune response, wherein the liposomes are targeted to type 2 dendritic cells.\",\n \"59.The method of claim 58, wherein the type 2 dendritic cells are immature type 2 dendritic cells.\",\n \"60.A method of enhancing an immune response comprising administering to the individual liposomes, wherein the liposomes comprise one or more antigens, wherein the liposomes are modified with surface-bound molecules that target the liposomes to mature type 1 dendritic cells.\",\n \"61.A method of modifying liposomes comprising packaging one or more antigens into a lipopsome, wherein the liposome is modified with surface-bound molecules that target the liposome to type 2 dendritic cells or mature type 1 dendritic cells.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Dendritic cells have specialized capacity to present peptide antigens to T-cells and regulate the initiation of the immune response (Hart, D. N. (1997) “Dendritic cells: unique leukocyte populations which control the primary immune response.” Blood 90: 3245-3287; Bancereau, J. and R. M. Steinman (1998) “Dendritic cells and the control of immunity.” Nature 392: 245-252).\",\n \"There is significant heterogeneity within the dendritic cell lineage.\",\n \"Dendritic cells develop from hematopoietic progenitor “stem” cells under the influence of cytokines which act during early myelo-monocytic differentiation (Peters, J. H., J. Ruppert, et al.\",\n \"(1991) “Differentiation of human monocytes into CD14 negative accessory cells: do dendritic cells derive from the monocytic lineage?” Pathobiology 59: 122-126;Santiago, S. F., E. Belilos, et al.\",\n \"(1992) “TNF in combination with GM-CSF enhances the differentiation of neonatal cord blood stem cells into dendritic cells and macrophages.” J Leukoc Biol 52(3): 274-81; Grouard, G., M. C. Rissoan, et al.\",\n \"(1997) “The enigmatic plasmacytoid T cells develop into dendritic cells with interleukin (IL)-3 and CD40-ligand.” J Exp Med 185: 1101-11; Olweus, J., A. BitMansour, et al.\",\n \"(1997) “Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.” Proc Natl Acad Sci USA 94: 12551-6; Rissoan, M. C., V. Soumelis, et al.\",\n \"(1999) “Reciprocal control of T helper cell and dendritic cell differentiation.” Science 283: 1183-6; Siegal, F. P., N. Kadowaki, et al.\",\n \"(1999) “The nature of the principal type 1 interferon-producing cells in human blood.” Science 284: 1835-7; Bergthiier, R., C. Martinon-Ego, et al.\",\n \"(2000) “A two step-culture method starting with early growth factors permits enhanced production of functional dendritic cells from murine splenocytes.” Immunological Methods 239: 95-107; Ferlazzo, G., J. Klein, et al.\",\n \"(2000) “Dendritic cells generated from CD34+ progenitor cells with flt3 ligand, c-kit ligand, GM-CSF, IL4, and TNF-alpha are functional antigen-presenting cells resembling monocyte-derived dendritic cells.” Journal of Immunotherapy 23: 48-58).\",\n \"Phenotypic and functional analyses have demonstrated two general types of dendritic cells, type 1 (DC 1) and type 2 (DC2), that differ in surface marker expression as well as their functional effect on cognate T-cells.\",\n \"Studies of cytokines used to mobilize peripheral blood hematopoietic progenitors have indicated increased numbers of DC I after administration of Flt3 and GM-CSF, while increased numbers of DC2 have been noted after treatment with G-CSF (Arpinati, M., C. L. Green, et al.\",\n \"(2000) “Granulocyte-colony stimulating factor mobilizes T helper-2-inducing dendritic cells.” Blood 95: 2484-2490).\",\n \"Interaction of DC1 with T-cells results in polarization of the T-cells towards Th1 and Tc 1 immune responses characterized by the generation of cytotoxic effectors and production of IL12, TNF and IFN-gamma.\",\n \"In contrast, interaction of DC2 with T-cells leads to T-cells that facilitate humoral immune responses, secrete IL4 and IL4, and suppress cytotoxic Th1 immune responses.\",\n \"DC have an important role in the anti-tumor effect of autologous and allogeneic transplantation.\",\n \"Antigen primed DC in have been used with promising results in adoptive vaccination against tumors (Mayordomo, J. I., T. Zorina, et al.\",\n \"(1995) “Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity.” Nature Medicine 1(12): 1297-302; Reichardt, V. L., C. Y. Okada, et al.\",\n \"(1999) “Idiotype vaccination using dendritic cells after autologous peripheral blood stem cell transplantation for multiple myeloma—a feasibility study.” Blood 93(7): 2411-9).\",\n \"DC1 promote Th1 immune responses in responding CD4 + T-cells characterized by enhanced INF-gamma, TNF, and IL-12 synthesis.\",\n \"DC2 promote Th2 responses in CD4+ T-cells characterized by IL4 and IL10 synthesis, and inhibition of INF-gamma and TNF production in cognate T-cells (Rissoan, M. C., V. Soumelis, et al.\",\n \"(1999) “Reciprocal control of T helper cell and dendritic cell differentiation.” Science 283: 1183-6).\",\n \"Bone marrow contains monocytes and CD86 + , CD34 + progenitor cells which can differentiate into DC1 in the presence of TNF and GM-CSF (Sallusto, F. and A. Lanzavecchia (1994) “Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor a.” Journal of Experimental Medicine 179: 1109-1118; Ye, Z., A. P. Gee, et al.\",\n \"(1996) “In vitro expansion and characterization of dendritic cells derived from human bone marrow CD34+ cells.” Bone Marrow Transplantation 18(5): 997-1008; Ryncarz, R. E. and C. Anasetti (1998) “Expression of CD86 on human marrow CD34(+) cells identifies immunocompetent committed precursors of macrophages and dendritic cells.” Blood 91(10): 3892-900), as well as CD123 bright DC progenitors which differentiate into DC2 in the presence of IL3 (Olweus, J., A. BitMansour, et al.\",\n \"(1997) “Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.” Proc Natl Acad Sci U S A 94: 12551-6).\",\n \"Most studies involving hematopoietic progenitor cell transplantation to date have examined the role of DC1 as a method of adoptive immunotherapy for cancer following autologous HPC transplantation (Mayordomo, J. I., T. Zorina, et al.\",\n \"(1995) “Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity.” Nature Medicine 1(12): 1297-302).\",\n \"One recent report, using a murine transplant model, indicated that only host CD11c+DC (DC 1) were necessary for the development of acute GvHD (Shlomchik, W. D., M. S. Couzens, et al.\",\n \"(1999) “Prevention of graft versus host disease by inactivation of host antigen-presenting cells.” Science 285: 412-5).\",\n \"Donor DC did not appear to efficiently present host antigens via cross-priming in a way that led to the initiation of GvHD (Shlomchik, W. D., M. S. Couzens, et al.\",\n \"(1999) “Prevention of graft versus host disease by inactivation of host antigen-presenting cells.” Science 285: 412-5).\",\n \"In this study, the possible effect of donor DC on inhibiting GvHD was not examined.\",\n \"The potential role of increased numbers of donor DC2 in regulating GvHD after G-CSF mobilized allogeneic peripheral stem cell transplants has recently been recognized (Arpinati, M., C. L. Green, et al.\",\n \"(2000).\",\n \"“Granulocyte-colony stimulating factor mobilizes T helper-2-inducing dendritic cells.” Blood 95: 2484-2490).\"\n ],\n [\n \"<SOH> BRIEF SUMMARY OF THE INVENTION <EOH>In accordance with the purposes of this invention, as embodied and broadly described herein, this invention, in one aspect, relates to a method of inducing immunological tolerance in an individual, where the method comprises administering to the individual type 2 dendritic cells or immature type 1 dendritic cells, where the dendritic cells have been incubated with one or more antigens.\",\n \"In another aspect, the invention relates to a method of reducing immune responsiveness to particular antigens, where the method comprises administering to the individual liposomes, where the liposomes contain one or more antigens, and where the liposomes are modified with surface-bound molecules that target the liposomes to either type 2 dendritic cells or mature type 1 dendritic cells.\",\n \"In another aspect, the invention relates to a method of enhancing immune responsiveness, where the method comprises administering to the individual liposomes, where the liposomes contain one or more antigens, where the liposomes are modified with surface-bound molecules that target the liposomes to mature type I dendritic cells.\",\n \"In these various aspects, the antigens can be, for example, carbohydrates, nucleic acids, peptides, lipids, or a combination of two or more of carbohydrates, nucleic acids, peptides, and lipids.\",\n \"In particular, the antigens can be MHC, QA, HIV gag, pol, and env (DNA or protein), rheumatoid factor, ICA 89, peripherin, carboxypeptidase H, glutamic acid decarboxylase (GAD), AHNAK, mylein basic protein, retinal S antigen, galactomannoprotein, neuraminidase, influenza matrix protein M1, CMV env proteins pp60, ds DNA, thyroglobulin, insulin, pancreatic islet beta cell antigens, MART-1 (melanoma protein), tyrosinase-related protein 2 (TRP2), melanoma cell lysate, MAGE-A3, allogeneic cell lysate, HLA antigens (protein or DNA), CD31 (protein or DNA), aspergillus chitin , pancreatic carcinoma cell line Panc-1 lysate, ErbB-2/neu, human epithelial cell mucin (Muc-1), Ha-ras oncogene product, p53, carcino-embryonic antigen (CEA), raf oncogene product, gp100/pmel17, GD2, GD3, GM2, TF, sTn, MAGE-1, MAGE-3, BAGE, GAGE, tyrosinase, gp75, Melan-A/Mart-1, gp100, HER2/neu, EBV-LMP 1 & 2, HPV-F4, 6, 7, prostate-specific antigen (PSA), prostate specific membrane antigen (PSMA), HPV-16, MUM, alpha-fetoprotein (AFP), CO17-1A, GA733, gp72, p53, ras oncogene product, HPV E7, Wilm's tumor antigen-1, telomerase, melanoma gangliosides, malignant B cell antigen receptor, malignant B cell immunoglobulin idiotype, variable region of an immunoglobulin, hypervariable region or complementarity determining region (CDR) of a variable region of an immunoglobulin, malignant T cell receptor (TCR), variable region of a TCR, hypervariable region of a TCR, or a combination.\",\n \"The antigens also can be auto-immune antigens, such as antigens involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.\",\n \"The antigens can be antigens involved in allograft rejection, such that the induced tolerance reduces the allograft rejection.\",\n \"The antigens also can be antigens involved in graft rejection of hematopoietic stem cell transplants, such that the induced tolerance reduces the graft rejection.\",\n \"The antigens also can be antigens involved in graft verses host disease.\",\n \"For those aspects of the disclosed method involving induction of immunological tolerance, the immunological tolerance can reduce an auto-immune response, such as an auto-immune response involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.\",\n \"The immunological tolerance can reduce an allergic immune response, such as an allergic immune response is involved in asthma, gluten allergy, and atopic dermatitis.\",\n \"The graft rejection immune response can comprise graft rejection and/or complications associated with graft rejection.\",\n \"The immunological tolerance can reduce graft verses host immune response or graft rejection immune response.\",\n \"The individual can be an allograft recipient, such as a hematopoietic stem cell recipient.\",\n \"The disclosed method of administration of antigen incubated dendritic cells can have additional features and aspects.\",\n \"For example, the incubation step can occur or be performed ex vivo, and type 2 dendritic cells can be isolated prior to the incubation by selection with an anti-BDCA-2 antibody.\",\n \"In the case of type 2 dendritic cells, the dendritic cells can be further purified selection for CD123+ cells and/or removal of CD11c− cells.\",\n \"Thus, in one aspect of the disclosed method, the type 2 dendritic cells are isolated prior to the incubation with antigen by selection with an anti-BDCA-2 antibody, selection for CD 123+ cells, and removal of CD11c− cells.\",\n \"Alternatively, type 1 dendritic cells can be isolated with an anti-BDCA-1 antibody.\",\n \"Type 1 dendritic cells can be further purified by removal of BDCA-2+ and/or CD123+ cells.\",\n \"The disclosed method of administration of liposomes targeted to dendritic cells can have additional features and aspects.\",\n \"For example, the surface-bound molecule can be a monoclonal antibody, or a peptide high-affinity ligand.\",\n \"In some embodiments, the surface-bound molecules can be specific for CD11c+, BDCA-1, or both.\",\n \"This targets mature type 1 dendritic cells In other embodiments, the surface-bound molecules can be specific for CD123, BDCA-2, BDCA-4, or a combination.\",\n \"This targets type 2 dendritic cells.\",\n \"The disclosed method can have a variety of effects and objects.\",\n \"For example, the liposomes can enhance anti-tumor immune responses, can enhance anti-cancer immune responses, can reduce an auto-immune response.\",\n \"In the latter case, the auto-immune response can be involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.\",\n \"The liposomes can reduce an allergic immune response, such as an allergic immune response is involved in asthma, gluten allergy, and atopic dermatitis.\",\n \"The liposomes can reduce graft verses host immune responses in individuals who are hematopoietic allograft recipients, such as hematopoietic stem cell transplant recipients.\",\n \"The liposomes can reduce graft rejection immune responses such as those that are complications associated with organ allograft transplants.\",\n \"In another aspect, the invention relates to a method of modifying liposomes, where the method comprises packaging one or more antigens into a lipopsome, where the liposome is modified with surface-bound molecules that target the liposome to either type 2 dendritic cells or immature type 1 dendritic cells.\",\n \"Additional advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.\",\n \"The advantages of the invention will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims.\",\n \"It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.\"\n ],\n [\n \"CROSS-REFERENCE TO RELATED APPLICATIONS This application claims benefit of U.S.\",\n \"Provisional Application No.\",\n \"60/289,625, filed May 8, 2001, which application is hereby incorporated herein by reference in its entirety.\",\n \"FIELD OF THE INVENTION The disclosed invention is generally in the field of altering immune responses and specifically in the field of altering immune responses via dendritic cells.\",\n \"BACKGROUND OF THE INVENTION Dendritic cells have specialized capacity to present peptide antigens to T-cells and regulate the initiation of the immune response (Hart, D. N. (1997) “Dendritic cells: unique leukocyte populations which control the primary immune response.” Blood 90: 3245-3287; Bancereau, J. and R. M. Steinman (1998) “Dendritic cells and the control of immunity.” Nature 392: 245-252).\",\n \"There is significant heterogeneity within the dendritic cell lineage.\",\n \"Dendritic cells develop from hematopoietic progenitor “stem” cells under the influence of cytokines which act during early myelo-monocytic differentiation (Peters, J. H., J. Ruppert, et al.\",\n \"(1991) “Differentiation of human monocytes into CD14 negative accessory cells: do dendritic cells derive from the monocytic lineage?” Pathobiology 59: 122-126;Santiago, S. F., E. Belilos, et al.\",\n \"(1992) “TNF in combination with GM-CSF enhances the differentiation of neonatal cord blood stem cells into dendritic cells and macrophages.” J Leukoc Biol 52(3): 274-81; Grouard, G., M. C. Rissoan, et al.\",\n \"(1997) “The enigmatic plasmacytoid T cells develop into dendritic cells with interleukin (IL)-3 and CD40-ligand.” J Exp Med 185: 1101-11; Olweus, J., A. BitMansour, et al.\",\n \"(1997) “Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.” Proc Natl Acad Sci USA 94: 12551-6; Rissoan, M. C., V. Soumelis, et al.\",\n \"(1999) “Reciprocal control of T helper cell and dendritic cell differentiation.” Science 283: 1183-6; Siegal, F. P., N. Kadowaki, et al.\",\n \"(1999) “The nature of the principal type 1 interferon-producing cells in human blood.” Science 284: 1835-7; Bergthiier, R., C. Martinon-Ego, et al.\",\n \"(2000) “A two step-culture method starting with early growth factors permits enhanced production of functional dendritic cells from murine splenocytes.” Immunological Methods 239: 95-107; Ferlazzo, G., J. Klein, et al.\",\n \"(2000) “Dendritic cells generated from CD34+ progenitor cells with flt3 ligand, c-kit ligand, GM-CSF, IL4, and TNF-alpha are functional antigen-presenting cells resembling monocyte-derived dendritic cells.” Journal of Immunotherapy 23: 48-58).\",\n \"Phenotypic and functional analyses have demonstrated two general types of dendritic cells, type 1 (DC 1) and type 2 (DC2), that differ in surface marker expression as well as their functional effect on cognate T-cells.\",\n \"Studies of cytokines used to mobilize peripheral blood hematopoietic progenitors have indicated increased numbers of DC I after administration of Flt3 and GM-CSF, while increased numbers of DC2 have been noted after treatment with G-CSF (Arpinati, M., C. L. Green, et al.\",\n \"(2000) “Granulocyte-colony stimulating factor mobilizes T helper-2-inducing dendritic cells.” Blood 95: 2484-2490).\",\n \"Interaction of DC1 with T-cells results in polarization of the T-cells towards Th1 and Tc 1 immune responses characterized by the generation of cytotoxic effectors and production of IL12, TNF and IFN-gamma.\",\n \"In contrast, interaction of DC2 with T-cells leads to T-cells that facilitate humoral immune responses, secrete IL4 and IL4, and suppress cytotoxic Th1 immune responses.\",\n \"DC have an important role in the anti-tumor effect of autologous and allogeneic transplantation.\",\n \"Antigen primed DC in have been used with promising results in adoptive vaccination against tumors (Mayordomo, J. I., T. Zorina, et al.\",\n \"(1995) “Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity.” Nature Medicine 1(12): 1297-302; Reichardt, V. L., C. Y. Okada, et al.\",\n \"(1999) “Idiotype vaccination using dendritic cells after autologous peripheral blood stem cell transplantation for multiple myeloma—a feasibility study.” Blood 93(7): 2411-9).\",\n \"DC1 promote Th1 immune responses in responding CD4+ T-cells characterized by enhanced INF-gamma, TNF, and IL-12 synthesis.\",\n \"DC2 promote Th2 responses in CD4+ T-cells characterized by IL4 and IL10 synthesis, and inhibition of INF-gamma and TNF production in cognate T-cells (Rissoan, M. C., V. Soumelis, et al.\",\n \"(1999) “Reciprocal control of T helper cell and dendritic cell differentiation.” Science 283: 1183-6).\",\n \"Bone marrow contains monocytes and CD86+, CD34+ progenitor cells which can differentiate into DC1 in the presence of TNF and GM-CSF (Sallusto, F. and A. Lanzavecchia (1994) “Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor a.” Journal of Experimental Medicine 179: 1109-1118; Ye, Z., A. P. Gee, et al.\",\n \"(1996) “In vitro expansion and characterization of dendritic cells derived from human bone marrow CD34+ cells.” Bone Marrow Transplantation 18(5): 997-1008; Ryncarz, R. E. and C. Anasetti (1998) “Expression of CD86 on human marrow CD34(+) cells identifies immunocompetent committed precursors of macrophages and dendritic cells.” Blood 91(10): 3892-900), as well as CD123brightDC progenitors which differentiate into DC2 in the presence of IL3 (Olweus, J., A. BitMansour, et al.\",\n \"(1997) “Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.” Proc Natl Acad Sci U S A 94: 12551-6).\",\n \"Most studies involving hematopoietic progenitor cell transplantation to date have examined the role of DC1 as a method of adoptive immunotherapy for cancer following autologous HPC transplantation (Mayordomo, J. I., T. Zorina, et al.\",\n \"(1995) “Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity.” Nature Medicine 1(12): 1297-302).\",\n \"One recent report, using a murine transplant model, indicated that only host CD11c+DC (DC 1) were necessary for the development of acute GvHD (Shlomchik, W. D., M. S. Couzens, et al.\",\n \"(1999) “Prevention of graft versus host disease by inactivation of host antigen-presenting cells.” Science 285: 412-5).\",\n \"Donor DC did not appear to efficiently present host antigens via cross-priming in a way that led to the initiation of GvHD (Shlomchik, W. D., M. S. Couzens, et al.\",\n \"(1999) “Prevention of graft versus host disease by inactivation of host antigen-presenting cells.” Science 285: 412-5).\",\n \"In this study, the possible effect of donor DC on inhibiting GvHD was not examined.\",\n \"The potential role of increased numbers of donor DC2 in regulating GvHD after G-CSF mobilized allogeneic peripheral stem cell transplants has recently been recognized (Arpinati, M., C. L. Green, et al.\",\n \"(2000).\",\n \"“Granulocyte-colony stimulating factor mobilizes T helper-2-inducing dendritic cells.”Blood 95: 2484-2490).\",\n \"BRIEF SUMMARY OF THE INVENTION In accordance with the purposes of this invention, as embodied and broadly described herein, this invention, in one aspect, relates to a method of inducing immunological tolerance in an individual, where the method comprises administering to the individual type 2 dendritic cells or immature type 1 dendritic cells, where the dendritic cells have been incubated with one or more antigens.\",\n \"In another aspect, the invention relates to a method of reducing immune responsiveness to particular antigens, where the method comprises administering to the individual liposomes, where the liposomes contain one or more antigens, and where the liposomes are modified with surface-bound molecules that target the liposomes to either type 2 dendritic cells or mature type 1 dendritic cells.\",\n \"In another aspect, the invention relates to a method of enhancing immune responsiveness, where the method comprises administering to the individual liposomes, where the liposomes contain one or more antigens, where the liposomes are modified with surface-bound molecules that target the liposomes to mature type I dendritic cells.\",\n \"In these various aspects, the antigens can be, for example, carbohydrates, nucleic acids, peptides, lipids, or a combination of two or more of carbohydrates, nucleic acids, peptides, and lipids.\",\n \"In particular, the antigens can be MHC, QA, HIV gag, pol, and env (DNA or protein), rheumatoid factor, ICA 89, peripherin, carboxypeptidase H, glutamic acid decarboxylase (GAD), AHNAK, mylein basic protein, retinal S antigen, galactomannoprotein, neuraminidase, influenza matrix protein M1, CMV env proteins pp60, ds DNA, thyroglobulin, insulin, pancreatic islet beta cell antigens, MART-1 (melanoma protein), tyrosinase-related protein 2 (TRP2), melanoma cell lysate, MAGE-A3, allogeneic cell lysate, HLA antigens (protein or DNA), CD31 (protein or DNA), aspergillus chitin, pancreatic carcinoma cell line Panc-1 lysate, ErbB-2/neu, human epithelial cell mucin (Muc-1), Ha-ras oncogene product, p53, carcino-embryonic antigen (CEA), raf oncogene product, gp100/pmel17, GD2, GD3, GM2, TF, sTn, MAGE-1, MAGE-3, BAGE, GAGE, tyrosinase, gp75, Melan-A/Mart-1, gp100, HER2/neu, EBV-LMP 1 & 2, HPV-F4, 6, 7, prostate-specific antigen (PSA), prostate specific membrane antigen (PSMA), HPV-16, MUM, alpha-fetoprotein (AFP), CO17-1A, GA733, gp72, p53, ras oncogene product, HPV E7, Wilm's tumor antigen-1, telomerase, melanoma gangliosides, malignant B cell antigen receptor, malignant B cell immunoglobulin idiotype, variable region of an immunoglobulin, hypervariable region or complementarity determining region (CDR) of a variable region of an immunoglobulin, malignant T cell receptor (TCR), variable region of a TCR, hypervariable region of a TCR, or a combination.\",\n \"The antigens also can be auto-immune antigens, such as antigens involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.\",\n \"The antigens can be antigens involved in allograft rejection, such that the induced tolerance reduces the allograft rejection.\",\n \"The antigens also can be antigens involved in graft rejection of hematopoietic stem cell transplants, such that the induced tolerance reduces the graft rejection.\",\n \"The antigens also can be antigens involved in graft verses host disease.\",\n \"For those aspects of the disclosed method involving induction of immunological tolerance, the immunological tolerance can reduce an auto-immune response, such as an auto-immune response involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.\",\n \"The immunological tolerance can reduce an allergic immune response, such as an allergic immune response is involved in asthma, gluten allergy, and atopic dermatitis.\",\n \"The graft rejection immune response can comprise graft rejection and/or complications associated with graft rejection.\",\n \"The immunological tolerance can reduce graft verses host immune response or graft rejection immune response.\",\n \"The individual can be an allograft recipient, such as a hematopoietic stem cell recipient.\",\n \"The disclosed method of administration of antigen incubated dendritic cells can have additional features and aspects.\",\n \"For example, the incubation step can occur or be performed ex vivo, and type 2 dendritic cells can be isolated prior to the incubation by selection with an anti-BDCA-2 antibody.\",\n \"In the case of type 2 dendritic cells, the dendritic cells can be further purified selection for CD123+ cells and/or removal of CD11c− cells.\",\n \"Thus, in one aspect of the disclosed method, the type 2 dendritic cells are isolated prior to the incubation with antigen by selection with an anti-BDCA-2 antibody, selection for CD 123+ cells, and removal of CD11c− cells.\",\n \"Alternatively, type 1 dendritic cells can be isolated with an anti-BDCA-1 antibody.\",\n \"Type 1 dendritic cells can be further purified by removal of BDCA-2+ and/or CD123+ cells.\",\n \"The disclosed method of administration of liposomes targeted to dendritic cells can have additional features and aspects.\",\n \"For example, the surface-bound molecule can be a monoclonal antibody, or a peptide high-affinity ligand.\",\n \"In some embodiments, the surface-bound molecules can be specific for CD11c+, BDCA-1, or both.\",\n \"This targets mature type 1 dendritic cells In other embodiments, the surface-bound molecules can be specific for CD123, BDCA-2, BDCA-4, or a combination.\",\n \"This targets type 2 dendritic cells.\",\n \"The disclosed method can have a variety of effects and objects.\",\n \"For example, the liposomes can enhance anti-tumor immune responses, can enhance anti-cancer immune responses, can reduce an auto-immune response.\",\n \"In the latter case, the auto-immune response can be involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.\",\n \"The liposomes can reduce an allergic immune response, such as an allergic immune response is involved in asthma, gluten allergy, and atopic dermatitis.\",\n \"The liposomes can reduce graft verses host immune responses in individuals who are hematopoietic allograft recipients, such as hematopoietic stem cell transplant recipients.\",\n \"The liposomes can reduce graft rejection immune responses such as those that are complications associated with organ allograft transplants.\",\n \"In another aspect, the invention relates to a method of modifying liposomes, where the method comprises packaging one or more antigens into a lipopsome, where the liposome is modified with surface-bound molecules that target the liposome to either type 2 dendritic cells or immature type 1 dendritic cells.\",\n \"Additional advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.\",\n \"The advantages of the invention will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims.\",\n \"It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is a graph of different cells and treatment versus counts per minute (cpm; representing proliferation of the cells).\",\n \"As described in the Example, 50,000 DC were cultured with 50,000 T-cells in the presence of allo-antigen or 1 μg/ml Con A.\",\n \"FIG.\",\n \"2 is a graph of responder to stimulator ratio versus counts per minute (cpm; representing cell proliferation).\",\n \"As described in the Example, serial dilutions of purified DC were prepared with 50,000, 25,000, 12,500, 6,250, 3125, 1500, or 75 cells added to culture wells in combination with 50,000 T-cells.\",\n \"DETAILED DESCRIPTION OF THE INVENTION The present invention may be understood more readily by reference to the following detailed description of preferred embodiments of the invention and the Example included therein and to the Figures and their previous and following description.\",\n \"It is to be understood that this invention is not limited to specific synthetic methods, specific preparative and analytical techniques unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary.\",\n \"It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.\",\n \"Immature DC may be immuno-suppressive, and decrease antigen specific immune responses.\",\n \"A recently published study in normal donors evaluated the ability of immature dendritic cells to augment T-cell activation after in vitro exposure to antigen (Dhodapkar, M. V., C. Y. Li, et al.\",\n \"(1994) “Clinical spectrum of clonal proliferations of T-large granular lymphocytes: a T-cell clonopathy of undetermined significance?” Blood 84(5): 1620-7).\",\n \"Immature dendritic cells were derived from peripheral blood monocytes and then exposed to the influenza peptide MP, and KLH.\",\n \"After return of these DCs to the host, a marked decline in the frequency of MP specific INF-gamma producing cells was noted, and persisted for 30 days.\",\n \"At the same time, the frequency of influenza specific T-cells was unchanged suggesting that the decrease in MP specific INF producing cells was related to DC exposure to the antigen, rather than a global reduction in influenza response.\",\n \"Also, this reduction in MP specific INF-gamma producing cells was paralleled by an increase in the frequency of IL-10 producing MP specific cells, and a reduction in the ability of CD8+ Mp specific cells to kill target cells that had been loaded with Mp peptides.\",\n \"There was no reduction in the overall frequency of MP specific T-cells after return of the immature DC's to account for this change in antigen response.\",\n \"Recently, a new paradigm has been developed in allogeneic hematopoietic cell transplantation, namely, that donor DC2 inhibit chronic GvHD and GvT responses among recipients of allogeneic HPC transplants.\",\n \"Based upon the naturally occurring biologic variation of the DC2 content in bone marrow grafts from 113 normal allogeneic sibling donors, it was found that recipients of larger numbers of DC2 experienced more relapse, while recipients of fewer DC2 had less relapse and more chronic GvHD post-transplant (Waller, E. K., H. Rosenthal, et al.\",\n \"(2001) “Larger numbers of CD4(bright) dendritic cells in donor bone marrow are associated with increased relapse after allogeneic bone marrow transplantation.” Blood 97(10): 2948-56).\",\n \"Patients received un-manipulated bone marrow from HLA-matched siblings for the treatment of lymphoma, leukemia, and multiple myeloma.\",\n \"The quantity of CD3+ and CD34+ cells and DC2 content in the bone marrow grafts were measured by flow cytometry and correlated with clinical outcomes post-transplant.\",\n \"When the study was initiated, a rare population of CD3−, CD4bright, CD8−, low side scatter, bone marrow cells was noted.\",\n \"These cells were clearly distinct from CD4+ T-cells and CD4lo monocytes.\",\n \"The CD3−, CD4bright are identical to DC2 (CD123+, CD11c−) subsequently described in peripheral blood, lymph node, thymus, and bone marrow (Grouard, G., M. C. Rissoan, et al.\",\n \"(1997) “The enigmatic plasmacytoid T cells develop into dendritic cells with interleukin (IL)-3 and CD40-ligand.” J Exp Med 185: 1101-11; Olweus, J., A. BitMansour, et al.\",\n \"(1997) “Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.” Proc Natl Acad Sci USA 94: 12551-6).\",\n \"For these and other reasons, it was realized that dendritic cells could be used to alter immune responses using the disclosed method.\",\n \"Further, it was realized that functional differences between DC1 and DC2 can be exploited using the disclosed method.\",\n \"A. Definitions As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.\",\n \"Thus, for example, reference to “a liposome” includes mixtures of two or more such liposomes, and the like.\",\n \"Ranges may be expressed herein as from “about” one particular value, and/or to “about” another particular value.\",\n \"When such a range is expressed, another embodimentincludes from the one particular value and/or to the other particular value.\",\n \"Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment.\",\n \"It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint.\",\n \"In this specification and in the claims which follow, reference will be made to a number of terms which shall be defined to have the following meanings: “Optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.\",\n \"“Dendritic cell” or “DC”, as used herein, refers to a mature antigen presenting cell,which is identified by the expression of one or more of the following markers on its cell surface: CD40, CD80, CD86, CD83 and HLA-DR, or a dendritic cell progenitor, as defined below, or both.\",\n \"“Dendritic cell progenitor”, “dendritic progenitor cells”, “dendritic precursor cell”, “dendritic cell precursor”, “DCP”, or “immature dendritic cell”, used interchangeably herein, means a hematopoietic cell that differentiates intro a mature DC.\",\n \"Type 1 Dendritic cell progenitors are typically identified by the expression of one or more of the following markers on its cell surface: CD11c, CD13, CD14, CD33, CD34, HLA_DR, or CD4.Type 2 Dendritic cell progenitors are typically identified by the expression of one or more of the following markers on its cell surface: CD123, CD45RA, CD34, CD36, BDCA-2, BDCA-4, HLA-DR or CD4.Dendritic cell progenitors typically lack expression of co-stimulatory molecules CD40, CD80, and CD86 as well as the activation marker CD83.Dendritic cell progenitors also lack expression of CD3, CD56, and CD20, markers associated with T-cell, NK cell, or B-cell lineages.\",\n \"“Mature dendritic cell or DC”, as used herein, refers to a mature antigen presenting cell, which is identified by the expression of one or more of the following markers on its cell surface CD54, CD40, CD80, CD86, CD83 and HLA-DR. “Type 1 dendritic cells or DC1” refer to mature dendritic cells or dendritic cell precursors that express CD11c and CD1b but lack expression of BDCA-2.“Type 2 dendritic cells or DC2” refer to mature dendritic cells or dendritic cell precursors that lack expression of CD11c and lack expression of CD1b but express high levels of CD123 and BDCA-2.“Hematopoietic system reconstituting cells” means a population of cells, preferably human, that possess the capability of dividing and producing progeny that include all of the formed cellular elements of the blood.\",\n \"Sources of hematopoietic system reconstituting cells can include bone marrow (both fetal and adult) and peripheral blood mononuclear cells (PBMC).\",\n \"“Donor” or “donor source” means the animal, preferably human, that is the naturalsource from which cells are originally removed.\",\n \"“Recipient” means the animal, typically human, into which cells or liposomes are transplanted.\",\n \"“Allogeneic” means that the recipient is not the natural source from which transplant cells have been removed.\",\n \"B.\",\n \"Components and Compositions Disclosed are the components to be used to prepare the disclosed compositions as well as the compositions themselves to be used within the methods disclosed herein.\",\n \"These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc.\",\n \"of these materials are disclosed that, while specific reference of each various individual and collective combinations and permutation of these compounds may not be explicitly disclosed, each is specifically contemplated and described herein.\",\n \"For example, if a particular liposome is disclosed and discussed and a number of modifications that can be made to a number of molecules including the liposome are discussed, specifically contemplated is each and every combination and permutation of liposomes and the modifications that are possible unless specifically indicated to the contrary.\",\n \"Thus, if a class of liposomes A, B, and C are disclosed as well as a class of antigens D, E, and F, and an example of a combination molecule (that is, for example, an antigen-loaded liposome) A-D is disclosed, then even if each is not individually recited, each combination is individually and collectively contemplated.\",\n \"This means, in this example, that combinations, A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F should be considered disclosed.\",\n \"Likewise, any subset or combination of these is also disclosed.\",\n \"Thus, for example, the sub-group of A-E, B-F, and C-E would be considered disclosed.\",\n \"This concept applies to all aspects of this application including, but not limited to, steps in methods of making and using the disclosed compositions.\",\n \"Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the disclosed methods.\",\n \"C. Dendritic Cells Dendritic cells (DC) are antigen presenting cells which are important in the initiation of immune responses.\",\n \"Based on in vitro culturing and characterization, DC are thought to represent a family of related cell types, perhaps from a single lineage.\",\n \"Progenitors of DC (DCP) can now be identified in many tissues, such as bone marrow and blood, based on the expression of certain cell surface markers.\",\n \"One type of precursor cell, the Langerhans cell, is found in the epidermis.\",\n \"Work on Langerhans cells established that these immature precursors are capable of antigen uptake, but do not demonstrate a significant ability to stimulate T cells until they differentiate into mature DC.\",\n \"A variety of markers have been identified that are useful in defining certain cell types of the dendritic cell family.\",\n \"For example, markers expressed on the surface of mature dendritic cells can include CD1a, CD11c, DEC-205 (mouse), CD4, CD33, CD208 (DC-LAMP), CD54, CD11b.\",\n \"HLA-DR, CD40, CD80, CD83, and CD86.Typically, the following markers are not found on the surface of mature dendritic cells: CD20, CD3, CD14, CD16, CD2, CD207 (Langerin), E-cadherin, Birbeck granules, CD56, and CD19.Cell surface markers present on type-I dendritic cell precursors include CD11c, CD13, CD14, CD33, CD34, and HLA-DR.\",\n \"Cell surface markers present on type-2 dendritic cell precursors include CD4, CD45RA, CD34, CD123, HLA-DR, and CD36, while the following markers are not found on the surface of dendritic cell precursors: CD20, CD3, CD16, CD2, CD40, CD80, CD83 and CD86 (see for example Caux et al.\",\n \"Intl.\",\n \"Immunology 1994 6(8): 1177-1185; Steptoe et al.\",\n \"J.\",\n \"Immunology 1997 159:5483-5491; Grouard et al.\",\n \"(J. Exp.\",\n \"Med.\",\n \"1997 185(6): 1101-1111; O'Doherty et al.\",\n \"Immunology 1994 82: 487-493; Galy et al., J. Immunotherapy 1998 21(2): 132-141; Strunk et al., J. Exp.\",\n \"Med.\",\n \"1997 185(6): 1131-1136; Olweus et al.\",\n \"Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci.\",\n \"USA 1997 94:12551-12556; Prignano et al., Intl.\",\n \"J. Dermatol.\",\n \"1998 37:116-119).\",\n \"By the appropriate use of these markers, dendritic cells can be distinguished from dendritic cell progenitors.\",\n \"In particular, dendritic cells are mature antigen presenting cells, which can be identified by the expression of one or more of the following markers on its cell surface: CD40, CD80, CD86, CD83 and HLA-DR. Further, unless the context indicates otherwise, a dendritic cell progenitor is a “dendritic cell.” Dendritic cell progenitors are hematopoietic cells which can be identified by the expression of one or more of the following markers on its cell surface: Type-1 dendritic cell progenitors—CD11c, CD13, CD14, CD33, CD34, and HLA-DR. Type-2 dendritic cell progenitors—CD4, CD45RA, CD34, CD123, HLA-DR, and CD36.1.Type 2 Dendritic Cells Type 2 Dendritic cells or “plasmacytoid” dendritic cells are characterized as hematopoietic cells that can present antigen in the context of both MHC class I and MHC class II.\",\n \"These cells are CD11b−, CD11c−, CD123bright, BDCA-2+, BDCA-4+, HLA-DR+, lineage-(CD3−, CD14−, CD19−, and CD56−).\",\n \"In addition to presenting antigen to T cells type 2 dendritic cells also secrete IL-4 which suppresses IL-12 secretion by T-cells and redirects T-cell immune responses from a TH1 response to TH2.Type 2 dendritic cells are largely responsible for presenting allergens to T cells.\",\n \"2.Type 1 Dendritic Cells Type 1 Dendritic cells are characterized as hematopoietic cells that can present antigen in the context of both MHC class I and MHC class II.\",\n \"These cells are CD11b+, CD11c+, CD123dim, BDCA-1+, HLA-DR+, lineage-(CD3−, CD14−, CD19−, and CD56−).\",\n \"In addition to presenting antigen to T cells upon antigen encounter and co-stimulation through CD40-CD40L interactions, type 1 dendritic cells also up regulate IL-12 and secrete IFN-γ which suppresses direct T-cell immune responses towards a TH1 response.\",\n \"Type 1 dendritic cells are largely responsible for presenting bacteria, viruses, and intracellular parasites to T cells.\",\n \"3.Isolation of Dendritic Cells Dendritic cells to be incubated with antigens can be isolated using any suitable techniques.\",\n \"Useful techniques involve selection or removal of cells based on the presence of cell surface markers.\",\n \"Many cells, including dendritic cells useful for the disclosed method, carry and lack unique sets of cell surface markers.\",\n \"Useful markers identifying dendritic cells are described above and elsewhere herein.\",\n \"Any one or a combination of these markers can be used in the isolation of dendritic cells.\",\n \"Particularly useful for the disclosed method are the markers CD11c, CD123 and BDCA-2.The presence of CD123 and BDCA-2, and the absence of CD11c identify a particularly useful dendritic cell subset: type 2 dendritic cells (DC2).\",\n \"Cells can be selected for retention via markers present on the desired cells.\",\n \"Cells can be selected for removal via markers not present on the desired cells.\",\n \"Thus, the cells remaining after cells are removed via a particular cell marker are enriched for cells lacking that cell marker.\",\n \"Both selection and removal of cells using cell markers can be accomplished using, for example, specific binding molecules targeted to the cell marker involved.\",\n \"Useful specific binding molecules for cell separation are antibodies direct to and/or specific for a cell surface marker.\",\n \"The targeted cells can be separated and/or sorted from cells lacking the marker by any of a variety of techniques.\",\n \"In general, such techniques make use of a tag component associated with the specific binding molecule.\",\n \"For example, the tag component can be a fluorescent label, a paramagnetic bead, or column matrix.\",\n \"Fluorescent labels are useful for cell separation based on the well-established technique of fluorescence activated cell sorting (FACS).\",\n \"Likewise, paramagnetic beads and the like and affinity columns derivatized with appropriate specific binding molecules can be used to separate cells.\",\n \"Examples of fluorescent labels that can be used as tag components include fluorescein isothiocyanate (FITC), 5,6-carboxymethyl fluorescein, Texas red, nitrobenz-2-oxa-1,3-diazol-4-yl (NBD), coumarin, dansyl chloride, rhodamine, amino-methyl coumarin (AMCA), Eosin, Erythrosin, BODIPY®, Cascade Blue®, Oregon Green®, pyrene, lissamine, xanthenes, acridines, oxazines, phycoerythrin, macrocyclic chelates of lanthanide ions such as quantum dye™, fluorescent energy transfer dyes, such as thiazole orange-ethidium heterodimer, and the cyanine dyes Cy3, Cy3.5, Cy5, Cy5.5 and Cy7.Examples of other specific fluorescent labels that can be used as tag components include 3-Hydroxypyrene 5,8,10-Tri Sulfonic acid, 5-Hydroxy Tryptamine (5-HT), Acid Fuchsin, Alizarin Complexon, Alizarin Red, Allophycocyanin, Aminocoumarin, Anthroyl Stearate, Astrazon Brilliant Red 4G, Astrazon Orange R, Astrazon Red 6B, Astrazon Yellow 7 GLL, Atabrine, Auramine, Aurophosphine, Aurophosphine G, BAO 9 (Bisaminophenyloxadiazole), BCECF, Berberine Sulphate, Bisbenzamide, Blancophor FFG Solution, Blancophor SV, Bodipy F1, Brilliant Sulphoflavin FF, Calcien Blue, Calcium Green, Calcofluor RW Solution, Calcofluor White, Calcophor White ABT Solution, Calcophor White Standard Solution, Carbostyryl, Cascade Yellow, Catecholamine, Chinacrine, Coriphosphine O, Coumarin-Phalloidin, CY3.18, CY5.18, CY7, Dans (1-Dimethyl Amino Naphaline 5 Sulphonic Acid), Dansa (Diamino Naphtyl Sulphonic Acid), Dansyl NH—CH3, Diamino Phenyl Oxydiazole (DAO), Dimethylamino-5-Sulphonic acid, Dipyrrometheneboron Difluoride, Diphenyl Brilliant Flavine 7GFF, Dopamine, Erythrosin ITC, Euchrysin, FIF (Formaldehyde Induced Fluorescence), Flazo Orange, Fluo 3, Fluorescamine, Fura-2, Genacryl Brilliant Red B, Genacryl Brilliant Yellow 10GF, Genacryl Pink 3G, Genacryl Yellow 5GF, Gloxalic Acid, Granular Blue, Haematoporphyrin, Indo-1, Intrawhite Cf Liquid, Leucophor PAF, Leucophor SF, Leucophor WS, Lissamine Rhodamine B200 (RD200), Lucifer Yellow CH, Lucifer Yellow VS, Magdala Red, Marina Blue, Maxilon Brilliant Flavin 10 GFF, Maxilon Brilliant Flavin 8 GFF, MPS (Methyl Green Pyronine Stilbene), Mithramycin, NBD Amine, Nitrobenzoxadidole, Noradrenaline, Nuclear Fast Red, Nuclear Yellow, Nylosan Brilliant Flavin E8G, Oxadiazole, Pacific Blue, Pararosaniline (Feulgen), Phorwite AR Solution, Phorwite BKL, Phorwite Rev, Phorwite RPA, Phosphine 3R, Phthalocyanine, Phycoerythrin R, Polyazaindacene Pontochrome Blue Black, Porphyrin, Primuline, Procion Yellow, Pyronine, Pyronine B, Pyrozal Brilliant Flavin 7GF, Quinacrine Mustard, Rhodamine 123, Rhodamine 5 GLD, Rhodamine 6G, Rhodamine B, Rhodamine B 200, Rhodamine B Extra, Rhodamine BB, Rhodamine BG, Rhodamine WT, Serotonin, Sevron Brilliant Red 2B, Sevron Brilliant Red 4G, Sevron Brilliant Red B, Sevron Orange, Sevron Yellow L, SITS (Primuline), SITS (Stilbene Isothiosulphonic acid), Stilbene, Snarf 1, sulpho Rhodamine B Can C, Sulpho Rhodamine G Extra, Tetracycline, Thiazine Red R, Thioflavin S, Thioflavin TCN, Thioflavin 5, Thiolyte, Thiozol Orange, Tinopol CBS, True Blue, Ultralite, Uranine B, Uvitex SFC, Xylene Orange, and XRITC.\",\n \"D. Antigens The disclosed methods make use of antigens to affect the immune response to those antigens.\",\n \"For example, both antigens that cause desirable immune responses, such as antigens derived from pathogens (to be used to induce a protective immune response to the pathogen), and antigens that cause undesirable immune responses, such as antigens involved in auto-immune diseases, allergic reactions and graft rejection.\",\n \"Any antigen of interest can be used.\",\n \"Useful antigens include those involved in an immune response and/or disease or condition.\",\n \"Particularly useful antigens are those that can cause an undesirable immune response.\",\n \"Examples include antigens involved in auto-immune disease, allograft rejection, allergic reactions, allergic immune responses.\",\n \"Useful antigens include those involved in auto-immune disease (referred to herein as auto-immune antigens), such as antigens involved in diabetes mellitus, multiple sclerosis, Chron's disease, inflammatory bowel disease, rheumatoid arthritis, thyroiditis, vitiligo, and/or systemic lupus erythematosis.\",\n \"Useful antigens also include those involved in allograft rejection, such as graft rejection of hematopoietic stem cell transplants.\",\n \"Particularly useful antigens include MHC, QA, HIV gag, pol, and env (DNA or protein), rheumatoid factor, ICA 89, peripherin, carboxypeptidase H, glutamic acid decarboxylase (GAD), AHNAK, mylein basic protein, retinal S antigen, galactomannoprotein, neuraminidase, influenza matrix protein M1, CMV env proteins pp60, ds DNA, thyroglobulin, insulin, pancreatic islet beta cell antigens, MART-1 (melanoma protein), tyrosinase-related protein 2 (TRP2), melanoma cell lysate, MAGE-A3, allogeneic cell lysate, HLA antigens (protein or DNA), CD31 (protein or DNA), aspergillus chitin, pancreatic carcinoma cell line Panc-1 lysate, ErbB-2/neu, human epithelial cell mucin (Muc-1), Ha-ras oncogene product, p53, carcino-embryonic antigen (CEA), raf oncogene product, gp100/pmel17, GD2, GD3, GM2, TF, sTn, MAGE-1, MAGE-3, BAGE, GAGE, tyrosinase, gp75, Melan-A/Mart-1, gp100, HER2/neu, EBV-LMP 1 & 2, HPV-F4, 6, 7, prostate-specific antigen (PSA), prostate specific membrane antigen (PSMA), HPV-16, MUM, alpha-fetoprotein (AFP), CO17-1A, GA733, gp72, p53, ras oncogene product, HPV E7, Wilm's tumor antigen-1, telomerase, melanoma gangliosides, malignant B cell antigen receptor, malignant B cell immunoglobulin idiotype, variable region of an immunoglobulin, hypervariable region or complementarity determining region (CDR) of a variable region of an immunoglobulin, malignant T cell receptor (TCR), variable region of a TCR, and hypervariable region of a TCR.\",\n \"For induction of an immune response against cancer or tumor cells, the antigens can be tumor antigens.\",\n \"Tumor antigens for use in the disclosed method can be any tumor antigen now known or later identified as a tumor antigen.\",\n \"The appropriate tumor antigen used depends on the tumor type being treated.\",\n \"For example, the tumor antigen can be, but is not limited to human epithelial cell mucin (Muc-1; a 20 amino acid core repeat for Muc-1 glycoprotein, present on breast cancer cells and pancreatic cancer cells), the Ha-ras oncogene product, p53, carcino-embryonic antigen (CEA), the raf oncogene product, gp100/pmel17, GD2, GD3, GM2, TF, sTn, tyrosinase-related protein 2 (TRP2), melanoma cell lysate, MAGE-A3, MAGE-1, MAGE-3, BAGE, GAGE, tyrosinase, gp75, Melan-A/Mart-1, gp100, HER2/neu, ErbB-2/neu, pancreatic carcinoma cell line Panc-1 lysate, EBV-LMP 1 & 2, HPV-F4, 6, 7, prostate-specific antigen (PSA), prostate specific membrane antigen (PSMA), HPV-16, MUM, alpha-fetoprotein (AFP), CO17-1A, GA733, gp72, p53, the ras oncogene product, HPV E7, Wilm's tumor antigen-1, telomerase, and melanoma gangliosides, as well as any other tumor antigens now known or identified in the future.\",\n \"Tumor antigens can be obtained following known procedures or are commercially available.\",\n \"Additionally, the tumor antigen of the present invention can be an antibody which can be produced by a B cell tumor (e.g., B cell lymphoma; B cell leukemia; myeloma) or the tumor antigen can be a fragment of such an antibody, which contains an epitope of the idiotype of the antibody.\",\n \"The epitope fragment can comprise as few as nine amino acids.\",\n \"For example, the tumor antigen of this invention can be a malignant B cell antigen receptor, a malignant B cell immunoglobulin idiotype, a variable region of an immunoglobulin, a hypervariable region or complementarity determining region (CDR) of a variable region of an immunoglobulin, a malignant T cell receptor (TCR), a variable region of a TCR and/or a hypervariable region of a TCR.\",\n \"Where the object of treatment is graft rejection, graft versus host disease, or auto-immune disease, the antigens can be prepared using cells from the source of the antigen(s) involved.\",\n \"For example, cells from the source of transplant cells can be used to obtain relevant antigens.\",\n \"As will be appreciated, the source cells contain antigens that are involved in graft rejection.\",\n \"Cells from the individual to be treated can be used to obtain relevant antigens in the case of auto-immune diseases.\",\n \"In general, whole cell preparations can be used.\",\n \"The cells can irradiated to kill the cells and the resulting treated cell mixture can be used as the antigens for incubation with dendritic cells of loading into liposomes targeted to dendritic cells.\",\n \"As a specific example, allo-antigen can be prepared by irradiating peripheral blood mononuclear cells from a third party donor to 25 Gy.\",\n \"Use of antigens prepared in this way is described in the Example.\",\n \"Useful antigens for altering immune responses in the context of transplants include MHC and QA antigens.\",\n \"Useful antigens for altering immune responses to influenza include neuraminidase, and influenza matrix protein M1.Useful antigens for altering immune responses to CMV include env proteins pp60.Useful antigens for altering immune responses in lupus include ds DNA and AHNAK.\",\n \"Useful antigens for altering immune responses in thyroditis include thyroglobulin.\",\n \"Useful antigens for altering immune responses in diabetes include insulin and pancreatic islet beta cell antigens.\",\n \"Useful antigens for altering immune responses to allografts include allogeneic cell lysate, HLA antigens (protein or DNA), and CD31 (protein or DNA).\",\n \"Useful antigens for altering auto-immune responses include rheumatoid factor, ICA 89, peripherin, carboxypeptidase H, and glutamic acid decarboxylase (GAD).\",\n \"Useful antigens for altering immune responses to fungal infections include aspergillus chitin and galactomannoprotein.\",\n \"Useful antigens for altering immune responses to pancreatic cancer include pancreatic carcinoma cell line Panc-1 lysate.\",\n \"Useful antigens for altering immune responses to prostate cancer include prostate specific antigen.\",\n \"Useful antigens for altering immune responses to colon cancer include carcioembryonic antigen.\",\n \"Useful antigens for altering immune responses to breast, renal, and/or lung cancer include ErbB-2/neu.\",\n \"Useful antigens for altering immune responses to HIV include HIV gag, pol, and env (DNA or protein).\",\n \"E. Liposomes The disclosed liposomes comprise liposomes comprising one or more antigens and one or more surface-bound molecules.\",\n \"The antigens are antigens of interest, such as antigens involved in undesirable immune responses.\",\n \"Liposomes comprising antigens are referred to herein as antigen-loaded liposomes.\",\n \"The surface-bound molecules are molecules that target the liposomes to dendritic cells.\",\n \"Liposomes containing surface-bound molecules are referred to herein as targeted liposomes.\",\n \"For convenience, the term liposome is used herein to refer to the base liposome, as well as to antigen-loaded and targeted liposomes.\",\n \"Generally, the context of the term indicates what is meant.\",\n \"Surface-bound molecules generally comprise a component that can bind to and/or interact with targeted cells.\",\n \"In the context of the disclosed method, the targeted cells are dendritic cells.\",\n \"Useful components that can bind to and/or interact with targeted cells are specific binding molecules.\",\n \"In the context of the disclosed method, the specific binding molecule is generally interacts with and/or is specific for a cell surface marker.\",\n \"Particularly useful targets for surface-bound molecules are CD11c, BDCA-1, or both, for targeting mature type 1 dendritic cells, and CD123, BDCA-2, BDCA-4, or a combination, for targeting type 2 dendritic cells.\",\n \"Liposomes are artificial structures primarily composed of phospholipid bilayers.\",\n \"Cholesterol and fatty acids may also be included in the bilayer construction.\",\n \"Liposomes may be loaded with compounds of interest, such as the disclosed antigens, and coated on the outer surface with surface-bound molecules, such as specific binding molecules.\",\n \"Liposomes, preferably unilamellar vesicles, can be made using established procedures that result in the loading of the interior compartment with antigens.\",\n \"The liposomes can also be associated with a surface-bound molecules, such as a specific binding molecule.\",\n \"The association may be direct or indirect.\",\n \"An example of a direct association is a liposome containing covalently bound antibodies on the surface of the phospholipid bilayer.\",\n \"An example of indirect association is a liposome containing covalently bound anti-antibody antibodies that can bind targeted antibodies.\",\n \"Liposome-forming compounds are generally well known as are the methods of their preparation.\",\n \"For example, any number of phospholipids or lipid compounds may be used to form the vesicle walls.\",\n \"Representative of such wall forming compounds are; phosphatidylcholine (hereinafter referred to as “PC”), both naturally occurring and synthetically prepared, phosphatidic acid (hereinafter referred to as “PA”), lysophosphatidylcholine, phosphatidylserine (hereinafter referred to as “PS”), phosphatidylethanolamine (hereinafter referred to as “PE”), sphingolipids, phosphatidyglycerol (hereinafter referred to as “PG”), spingomyelin, cardiolipin, glycolipids, gangliosides, cerebrosides and the like used either singularly or intermixed such as in soybean phospholipids.\",\n \"In addition, other lipids such as steroids, cholesterol, aliphatic amines such as long chain aliphatic amines and carboxylic acids, long chain sulfates and phosphates, dicetyl phosphate, butylated hydroxytoluene, tocophenol, retinol, and isoprenoid compounds may be intermixed with the phospholipid components to confer certain desired and known properties on the formed vesicles.\",\n \"In addition, synthetic phospholipids containing either altered aliphatic portions such as hydroxyl groups, branched carbon chains, cycloderivatives, aromatic derivatives, ethers, amides, polyunsaturated derivatives, halogenated derivatives or altered hydrophillic portions containing carbohydrate, glycol, phosphate, phosphonate, quaternary amine, sulfate, sulfonate, carboxy, amine, sulfhydryl, imidazole groups and combinations of such groups can be either substituted or intermixed with the above mentioned phospholipids and used in the disclosed method.\",\n \"It will be appreciated from the above that the chemical composition of the lipid component of the vesicles prepared by the method of the invention may be varied greatly without appreciable diminution of percentage capture although the size of the vesicle may be affected by the lipid composition.\",\n \"A convenient mixture we have used extensively and which is representative of lipid mixtures advantageously used in the method of the invention is composed of PS and PC, or PG and PC as identified above (advantageously at a 1:4 molar ratio in each instance).\",\n \"The PC, PG, PA and PE, may be derived from purified egg yolk.\",\n \"Saturated synthetic PC and PG, such as dipalmitoyl may also be used.\",\n \"Other amphipathic lipids that may be used, advantageously also at 1:4 molar ratios with PC, are gangliosides, globosides, fatty acids, stearylamine, long chain alcohols, and the like.\",\n \"The production and use of liposomes, including description of liposomes having useful properties and specialized components, are known.\",\n \"For example, various liposomes, their components and their method of production and use are described in U.S. Pat.\",\n \"No.\",\n \"6,296,870, U.S. Pat.\",\n \"No.\",\n \"6,241,999, U.S. Pat.\",\n \"No.\",\n \"6,214,388, U.S. Pat.\",\n \"No.\",\n \"5,354,853, U.S. Pat.\",\n \"No.\",\n \"4,900,556, U.S. Pat.\",\n \"No.\",\n \"4,708,861, U.S. Pat.\",\n \"No.\",\n \"4,235,871, and U.S. Pat.\",\n \"No.\",\n \"4,224,179, all of which are incorporated herein by reference for their description of he above subject matter.\",\n \"The class of liposomes described in U.S. Pat.\",\n \"No.\",\n \"6,214,388, which are designed to maximize internalization of liposomes by cells having cell surface markers of interest, are particularly useful in the disclosed method.\",\n \"1.Modifying Liposomes Disclosed is a method of modifying liposomes comprising loading liposomes with one or more antigens.\",\n \"The liposomes can also be modified by adding surface-bound molecules to the surface of the liposomes.\",\n \"The surface-bound molecules can target the liposomes to cells, such as dendritic cells.\",\n \"Techniques for loading liposomes and for producing liposome having surface-bound molecules are known.\",\n \"For example, liposomes can be loaded with antigens by forming the liposomes in the presence of the antigens (see, for example, liposome patents described above).\",\n \"Liposomes having surface-bound molecules can be produced, for example, by using lipids to which the surface-bound molecules are couple or otherwise attached.\",\n \"It is useful to couple surface-bound molecules at or near a polar or charged portion of the lipids.\",\n \"This should allow the surface-bound molecules to be exposed on the surface of the liposomes.\",\n \"F. Specific Binding Molecules A specific binding molecule is a molecule that interacts specifically with a particular molecule or moiety.\",\n \"In the context of the disclosed method, the molecule or moiety that interacts specifically with a specific binding molecule is generally a cell surface marker.\",\n \"Antibodies, either member of a receptor/ligand pair, synthetic polyamides (Dervan and Burli, Sequence-specific DNA recognition by polyamides.\",\n \"Curr Opin Chem Biol, 3(6):688-93 (1999); Wemmer and Dervan, Targeting the minor groove of DNA.\",\n \"Curr Opin Struct Biol, 7(3):355-61 (1997)), and other molecules with specific binding affinities are examples of specific binding molecules.\",\n \"Specific binding molecules are generally used to isolate useful dendritic cells and/or to target liposomes to dendritic cells as described elsewhere herein.\",\n \"A specific binding molecule that interacts specifically with a particular analyte is said to be specific for that analyte.\",\n \"For example, where the specific binding molecule is an antibody that associates with a particular antigen, the specific binding molecule is said to be specific for that antigen.\",\n \"The antigen is the analyte.\",\n \"A reporter molecule containing the specific binding molecule can also be referred to as being specific for a particular analyte.\",\n \"Specific binding molecules preferably are antibodies, ligands, binding proteins, receptor proteins, haptens, aptamers, carbohydrates, synthetic polyamides, peptide nucleic acids, or oligonucleotides.\",\n \"Preferred binding proteins are DNA binding proteins.\",\n \"Preferred DNA binding proteins are zinc finger motifs, leucine zipper motifs, helix-turn-helix motifs.\",\n \"These motifs can be combined in the same specific binding molecule.\",\n \"1.Antibodies Antibodies useful as specific binding molecules in the disclosed method are generally specific for cell surface markers.\",\n \"Numerous antibodies specific for various cell surface markers are known, and many are commercially available.\",\n \"Such antibodies can be used in the disclosed method and/or to produce components used in the disclosed method (such as for use as specific binding molecules).\",\n \"Useful antibodies can also be produced for use in the disclosed method.\",\n \"Techniques for antibody production are known, some of which are described below.\",\n \"Such techniques can be used to produce antibodies for use in the disclosed method.\",\n \"The term “antibodies” is used herein in a broad sense and includes both polyclonal and monoclonal antibodies.\",\n \"In addition to intact immunoglobulin molecules, also included in the term “antibodies” are fragments or polymers of those immunoglobulin molecules, and human or humanized versions of immunoglobulin molecules or fragments thereof, as long as they are chosen for their ability to interact with cell surface markers (or other antibody target), as described herein.\",\n \"The antibodies can be tested for their desired activity using the in vitro assays described herein, or by analogous methods.\",\n \"The term “monoclonal antibody” as used herein refers to an antibody obtained from a substantially homogeneous population of antibodies, that is, the individual antibodies within the population are identical except for possible naturally occurring mutations that may be present in a small subset of the antibody molecules.\",\n \"The monoclonal antibodies herein specifically include “chimeric” antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, as long as they exhibit the desired antagonistic activity (See, U.S. Pat.\",\n \"No.\",\n \"4,816,567 and Morrison et al., Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci.\",\n \"USA, 81:6851-6855 (1984)).\",\n \"Monoclonal antibodies can be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature, 256:495 (1975).\",\n \"In a hybridoma method, a mouse or other appropriate host animal is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent.\",\n \"Alternatively, the lymphocytes may be immunized in vitro, e.g., using the HIV Env-CD4-co-receptor complexes described herein.\",\n \"The monoclonal antibodies may also be made by recombinant DNA methods, such as those described in U.S. Pat.\",\n \"No.\",\n \"4,816,567 (Cabilly et al.).\",\n \"DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (for example, by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies).\",\n \"Libraries of antibodies or active antibody fragments can also be generated and screened using phage display techniques, for example, as described in U.S. Pat.\",\n \"No.\",\n \"5,804,440 to Burton et al.\",\n \"and U.S. Pat.\",\n \"No.\",\n \"6,096,441 to Barbas et al.\",\n \"In vitro methods are also suitable for preparing monovalent antibodies.\",\n \"Digestion of antibodies to produce fragments thereof, particularly, Fab fragments, can be accomplished using routine techniques known in the art.\",\n \"For instance, digestion can be performed using papain.\",\n \"Examples of papain digestion are described in WO 94/29348 published Dec. 22, 1994 and U.S. Pat.\",\n \"No.\",\n \"4,342,566.Papain digestion of antibodies typically produces two identical antigen binding fragments, called Fab fragments, each with a single antigen binding site, and a residual Fc fragment.\",\n \"Pepsin treatment yields a fragment that has two antigen combining sites and is still capable of cross-linking antigen.\",\n \"The fragments, whether attached to other sequences or not, can also include insertions, deletions, substitutions, or other selected modifications of particular regions or specific amino acids residues, provided the activity of the antibody or antibody fragment is not significantly altered or impaired compared to the non-modified antibody or antibody fragment.\",\n \"These modifications can provide for some additional property, such as to remove/add amino acids capable of disulfide bonding, to increase its bio-longevity, to alter its secretory characteristics, etc.\",\n \"In any case, the antibody or antibody fragment must possess a bioactive property, such as specific binding to its cognate antigen.\",\n \"Functional or active regions of the antibody or antibody fragment may be identified by mutagenesis of a specific region of the protein, followed by expression and testing of the expressed polypeptide.\",\n \"Such methods are readily apparent to a skilled practitioner in the art and can include site-specific mutagenesis of the nucleic acid encoding the antibody or antibody fragment.\",\n \"(Zoller, M. J. Curr.\",\n \"Opin.\",\n \"Biotechnol.\",\n \"3:348-354, 1992).\",\n \"As used herein, the term “antibody” or “antibodies” can also refer to a human antibody and/or a humanized antibody.\",\n \"G. Tag Components Tag components are molecules that can be associated with specific bindingmolecules and used to separate or sort cells.\",\n \"Useful tag components include fluorescent molecules and sortable components.\",\n \"Fluorescent molecules are particularly useful for fluorescence activated cell sorting (FACS).\",\n \"Sortable components include molecules and compositions that can be selectively bound or that can selectively interact with other molecules and compounds.\",\n \"Examples include beads or particles, such as paramagnetic bead, that can be separated based on a property of the bead.\",\n \"Thus, for example, cells associated with paramagnetic bead or particle can be selected or separated by magnetic sorting.\",\n \"Specific binding molecules are a useful form of tag component.\",\n \"Cells associated with such a specific binding molecule can be separated by associating the specific binding molecule with its binding partner (that is the molecule or moiety with which the specific binding molecule interacts specifically).\",\n \"Thus, for example, cells with which a specific binding molecule (as tag component) is associated can be selected or separated by passing the cells over a column derivatized with the binding partner of the specific binding molecule.\",\n \"H. Incubation of Dendritic Cells With Antigens Some forms of the disclosed methods involve or make use of dendritic cells that have been incubated with or exposed to one or more antigens.\",\n \"In particular, the disclosed method can use type 2 dendritic cells or CD11c−, CD123bright, BDCA-2+ dendritic cells that have been incubated with one or more antigens.\",\n \"Administration of such dendritic cells to an individual can induce immunological tolerance to the antigens.\",\n \"Alternatively, the disclosed method can use immature type 1 dendritic cells or CD11c−, CD123dim, BDCA-2− dendritic cells that have been incubated with one or more antigens.\",\n \"Administration of such type 1 dendritic cells to an individual can enhance immune responses to, for example, tumor antigens and antigens associated with infectious diseases.\",\n \"Dendritic cells are generally incubated with antigens ex vivo.\",\n \"In this context, ex vivo refers to treatment of the cells outside of the body from which they are derived or isolated.\",\n \"If ex vivo methods are employed, the cells can be maintained outside the body according to standard protocols well known in the art.\",\n \"The antigens can be incubated with the cells under any suitable conditions.\",\n \"Useful conditions include conditions useful for maintenance of the cells outside the body.\",\n \"As a specific example, preparations of antigens from irradiated cells can be incubated with dendritic cells.\",\n \"For example, 20,000 irradiated cells can be added to isolated dendritic cells and the cells incubated for 24 and 72 hours in a 5% CO2 atmosphere at 37° C. TNF can then be added (at, for example, 10 ng/ml) to mature and activate dendritic cell progenitors.\",\n \"Use of these incubation conditions is described in the Example.\",\n \"I.\",\n \"Administration of Dendritic Cells Dendritic cells exposed to antigen can be administered to individuals and animals using any suitable technique.\",\n \"In general, The treated cells can be infused per standard methods for the cell type.\",\n \"Standard methods are known for infusion of dendritic cells into a subject.\",\n \"In general, the disclosed dendritic cells can be administered by infusion, such as by intravenous infusion.\",\n \"A useful method of intraveneous infusion is to insert a indwelling catherter into a vein and attach to the catheter a flexible tubing, usually composed of plastic, and attach to the tubing a reservoir containing the substance to be infused.\",\n \"The reservoir may be a bag or syringe containing a liquid solution.\",\n \"The solution to be infused is transferred from the reservoir bag through the flexible tubing and indwelling catherter into the venous circulation of the recipient.\",\n \"The force to affect the transfer of the solution into the venous circulation may be gravity or compression of the bag or syringe.\",\n \"However, the dendritic cells may also be administered orally, parenterally (for example, intravenously), by intramuscular injection, by intraperitoneal injection, transdermally, sub-cutaneously, topically or the like.\",\n \"As used herein, “intravenous infusion” means delivery of the cells into a vein of the individual.\",\n \"Administration of the dendritic cells by inhalant can be through the nose or mouth via delivery by a spraying or droplet mechanism.\",\n \"J.\",\n \"Administration of Liposomes Liposomes can be administered to individuals and animals using any suitable technique.\",\n \"In general, the disclosed liposomes can be administered by infusion, such as by intravenous infusion.\",\n \"This is useful because the targeted dendritic cells are found in the blood.\",\n \"However, the liposomes may be administered orally, parenterally (for example, intravenously), by intramuscular injection, by intraperitoneal injection, transdermally, sub-cutaneously, topically or the like.\",\n \"As used herein, “intravenous infusion” means delivery of the liposomes into a vein of the individual.\",\n \"Administration of the liposomes by inhalant can be through the nose or mouth via delivery by a spraying or droplet mechanism.\",\n \"Delivery can also be directly to any area of the respiratory system (for example, lungs) via intubation.\",\n \"The exact amount of the liposomes required will vary from subject to subject, depending on the species, age, weight and general condition of the subject, the mode of administration and the like.\",\n \"Thus, it is not possible to specify an exact amount for every liposome.\",\n \"However, an appropriate amount can be determined by one of ordinary skill in the art using only routine experimentation given the teachings herein.\",\n \"K. Immune Responses An immune response refers to one or more effects produced by the immune system in reaction to one or more antigens.\",\n \"This includes, for example, immune system reactions to combinations of antigens; combinations of one or more antigens with other, non-antigenic components; and antigens in or on cells, viruses, liposomes or other compositions.\",\n \"As used herein, immune response refers to the anti-antigen function of the immune system and not to effects on and changes in the immune system that reduce the anti-antigen function of the immune system.\",\n \"For example, stimulation of neutralizing antibodies in response to exposure of an immune system to an antigen is an immune response, while induction of immunological tolerance is not an immune response as that term is used herein.\",\n \"It should be understood that this is the case even though immunological tolerance represents an effect produced by the immune system in reaction to one or more antigens.\",\n \"Adaptive immune response can largely be differentiated into two components humoral and cell-mediated.\",\n \"This division is based on whether the immune response is dominated by antibodies produced by plasma cells (a B cell lineage cell) or cytotoxic T cells.\",\n \"However, rarely are immune responses strictly the domain of either one of the components.\",\n \"Predominantly, the immune response is a combination of the two arms of the immune response having both T cell and B cell components.\",\n \"The dominance of either component is largely the result of regulatory CD4+ T cells, and the interaction of these cells with antigen in the context of dendritic cells.\",\n \"CD4 T cells can produce a myriad of functions such as cytokine release and direct cytotoxicity.\",\n \"It is these functions that determines whether the cell-mediated or humoral responses will dominate.\",\n \"Typically, CD4 T cells that stimulate a cell-mediate response are referred to as inflammatory CD4 T cells or TH1 cells.\",\n \"A TH1 response is characterized by IFN-γ, TNF, and IL 12 production from CD8 and CD4 T cells, IgG2a antibody production, and the generation of cytotoxic T cells.\",\n \"CD4 cells that induce a more humoral response are termed TH2.TH2 responses result in IL-4, IL5, and IL10 secretion and IgG1, IgA and IgE antibody production.\",\n \"The determination as to whether a CD4 cell will become a TH1 or TH2 cell is largely a factor of the environment that surround the cell at the time of activation.\",\n \"The environmental factors that control this are cytokines and the type of “non-professional” antigen-presenting cells or “professional” antigen-presenting cells (dendritic cells) that presents the antigen.\",\n \"Secreted IL-12 and IFN-γ in the presence of an uncommitted T cell can promote a TH1 response.\",\n \"Similarly, secreted IL-4 in the presence of an uncommitted T cell can promote a TH2 response.\",\n \"Additionally, these cytokines have the effect of suppressing the opposite response.\",\n \"The environment associated with the determination of the commitment to a TH1 or TH2 pathway also involves the cells that present antigen to CD4 T cells.\",\n \"CD4 T cells recognize peptide antigen in the context of MHC class II molecules presented by antigen presenting cells.\",\n \"These cells may be macrophage, mast cells, dendritic cells, or B cells.\",\n \"Depending on the cell type that presents the antigen the CD4 cell may be committed to either pathway.\",\n \"CD4 T cells activated by macrophage or type 1 dendritic cells are directed to induce a cell-mediated immune response.\",\n \"CD4 T cells activated by type 2 dendritic cells and mast cells are directed to induce a TH2 immune response.\",\n \"1.Reduction of Immune Responses Some embodiments of the disclosed methods result in reduction of an immune response.\",\n \"As used herein, reduction of an immune response refers to a lessening of one or more effects produced by the immune system in reaction to an antigen.\",\n \"Such reduction includes both reduction of an actual immune response or a reduction in an immune response that would otherwise have been expected in the absence of the reducing treatment.\",\n \"Reduction can include, but does not require, lessening of the immune response to an undetectable level.\",\n \"As used herein, suppression of an immune response refers to a lessening of one or more effects produced by the immune system in reaction to an antigen.\",\n \"Such suppression includes both lessening of an actual immune response or a lessening of an immune response that would otherwise have been expected in the absence of the suppressing treatment.\",\n \"Suppression can include, but does not require, lessening of the immune response to an undetectable level.\",\n \"2.Enhancement of Immune Responses Some embodiments of the disclosed methods result in enhancement of an immune response.\",\n \"As used herein, enhancement of an immune response refers to an increase of one or more effects produced by the immune system in reaction to an antigen.\",\n \"Such enhancement includes both enhancement of an actual immune response or an enhancement in an immune response that would otherwise not have been expected in the absence of the enhancing treatment.\",\n \"As used herein, generation of an immune response refers to the creation of one or more effects produced by the immune system (to a detectable level) in reaction to an antigen.\",\n \"Such generation includes creation of an immune response that would otherwise not have been expected in the absence of the generating treatment.\",\n \"L. Immunological Tolerance Some forms of the disclosed methods result in immunological tolerance to one or more antigens.\",\n \"As used herein, immunological tolerance refers to a physiological state in which the immune system does not react destructively against antigens.\",\n \"In effect, immunological tolerance is a lack of an immune response to an antigen.\",\n \"Mechanistically, immunological tolerance generally is the result of either deletion of reactive cells during development, such as “central tolerance” that occurs during deletion of auto-reactive T-cells during intrathymic development, or peripheral tolerance, that may occur during encounter of immune cells with antigen at peripheral lymphoid tissues in a manner that generates an anergis or unresponsive state.\",\n \"Immunological tolerance generally involves interruption of one or more steps involved in generation of an immune response.\",\n \"For example, individuals generally have immunological tolerance to self-antigens.\",\n \"This tolerance is a result of negative selection that takes place during T and B cell development in the thymus and bone marrow respectively.\",\n \"For T cells, this process involves the clonal deletion of T cells that recognize self-antigens.\",\n \"The deletion takes place in the thymus.\",\n \"Here CD4+CD8+ T cells are transitioning to either CD4−CD8+ or CD4+CD8− T cells.\",\n \"At this point in their development, the T cells are positively selected based on exposure and the subsequent ability to bind self-MHC.\",\n \"T cells that do not respond to the stimuli undergo activation induced cell death (AICD) and are deleted from the T cell repertoire, as being unable to recognize self-MHC would prevent the recognition of antigen since a TCR will only recognize antigen in the context of MHC.\",\n \"Those T cells that recognize self-MHC (that is, bind above a certain threshold) finish the differentiation to CD4−CD8+ or CD4+CD8− T cells.\",\n \"The surviving T cells are then negatively selected if they become activated by self-antigen being presented on self-MHC.\",\n \"This negative selection insures that the immune system won't respond to self and the responding T cells are deleted.\",\n \"Interestingly, foreign antigen present in the organism at the time of negative selection will also be seen as self.\",\n \"An example of this is the creation of Lymphocytic choriomeningitis virus (LCMV) carrier mice via neonatal infection.\",\n \"Adult mice infected with LCMV have a profound T cell response which can lead to meningitis due to immune pathology.\",\n \"Though a persistent infection can ensue, mice not dying via immune pathology will clear the infection.\",\n \"However, neonatal mice infected with LCMV will not generate an immune response and will become lifelong carriers of the virus showing no ill effects of infection or immune pathology.\",\n \"Similarly, mice receiving a non-lethal dose of irradiation will lose all T cells.\",\n \"Exposure to a foreign antigen prior to the reconstitution of the T cell population will result in animals that are tolerant to the foreign antigen.\",\n \"Though the mechanism of central tolerance through clonal deletion insures that the majority of T cells do not recognize self, there are tissue-specific antigens in the periphery that will not be presented to developing T cells in the thymus.\",\n \"Therefore, it is still possible to have T cells that react to self.\",\n \"However, the activation of a T cell is a two step process (1) the T cell must recognize antigen in the context of self-MHC and (2) the T cell must receive co-stimulatory signals in the form of CTLA-4-CD80/CD86, 41BB-41BBL, CD40-CD40L or CD28-B7.1/B7.2 interactions.\",\n \"Without both signals, a T cell will not become activated.\",\n \"In the situation of peripheral antigens, T cells specific for these self-antigens do not receive a co-stimulatory signal and therefore become anergic.\",\n \"In contrast to T cells, B cells do not recognize antigen in the context of MHC and therefore do not require a positive selection step.\",\n \"However, immature B cells do undergo negative selection within the bone marrow.\",\n \"Here, B cells with immunoglobulin (BCR) that recognizes self-antigens are clonally deleted.\",\n \"This was shown experimentally in mice transgenic for transmembrane bound hen-egg lysozyme (HEL) and transgenic for B cells specific for HEL.\",\n \"In these experiments, immature B cells specific for HEL can be detected, but no HEL specific mature B cells exist as they are deleted from the repertoire.\",\n \"Despite the deletion of self-specific cells in the bone marrow, there is still potential for mature B cells to exist that recognize soluble self-antigen or antigen only expressed in certain peripheral tissue.\",\n \"In the latter situation, B cells that recognize antigen expressed in peripheral tissue are deleted.\",\n \"However, B cells that recognize soluble antigen are not deleted.\",\n \"Instead these B cells internalize their antigen receptor and lose functionality.\",\n \"This loss of function is called anergy and results in a lack of response to an antigen despite the presence of specific cells.\",\n \"M. Pharmaceutically Acceptable Carriers The disclosed cells and liposomes can be used therapeutically in combination with a pharmaceutically acceptable carrier.\",\n \"Pharmaceutical carriers are known to those skilled in the art.\",\n \"These most typically would be standard carriers for administration of drugs to humans, including solutions such as sterile water, saline, and buffered solutions at physiological pH.\",\n \"Particularly useful pharmaceutical carriers are physiological solutions.\",\n \"In the case of the disclosed cells, media and solutions in which the cells are store, grown, maintained or.\",\n \"incubated are particularly useful as pharmaceutical carriers.\",\n \"Pharmaceutical compositions may include carriers, thickeners, diluents, buffers, preservatives, surface active agents and the like in addition to the cells or liposomes.\",\n \"Pharmaceutical compositions may also include one or more active ingredients such as antimicrobial agents, anti-inflammatory agents, anesthetics, and the like.\",\n \"It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope or spirit of the invention.\",\n \"Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein.\",\n \"It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being indicated by the following claims.\",\n \"EXAMPLE A.\",\n \"The Effect of DC2p Depletion on the Ability of the Cells in an HPC Product to Generate an Allo-Immune Response.\",\n \"The mononuclear cell fraction of the blood from a normal donor was collected by apheresis.\",\n \"Naïve CD45RA+ 62L+ T-cells were purified by FACS and frozen.\",\n \"The HPC product was depleted of DC2p using the BDCA2 antibody and the MACS immuno-affinity columns or purified DC1p and FACS isolated DC2p populations.\",\n \"The ability of the DC2p depleted HPC cells and the purified DCp2 and DC1p populations to generate cellular immune responses to allo-antigen was determined by adding allo-antigen to the DCp cells, maturing the DCp by the addition of IL3, GM-CSF, and TNF, then adding the previously collected naïve T-cells and measuring their proliferation in response to the allo-antigen (FIGS.\",\n \"1 and 2).\",\n \"The proliferative response of autologous T-cells to allo-antigen presented by purified DC2p was inhibited, while T-cells showed marked proliferation in response to allo-antigen presented by purified DC1p.\",\n \"Of note, T-cells had a strong proliferative response to the HPC graft that was depleted of the inhibitory DC2p population (stars, FIG.\",\n \"2).\",\n \"Procedural Details.\",\n \"Peripheral blood mononuclear cells were obtained form a normal donor by apheresis.\",\n \"Naïve CD3+, CD45RA+, CD62L+ naïve T-cells were isolated following staining with fluorescently labeled anti-CD3, anti-CD45RA, and anti 62L and high speed fluorescent activated cell sorting.\",\n \"DC1 (CD11c+, CD123dim, lineage-) and DC2 (CD11c−, CD123+, lineage-) populations were enriched from the apheresis product using immuno-affinity magnetic purification using antibodies to DC1 (BDCA-1) and DC2 (BDCA-2) then purified to >95% homogeneity by high speed cell sorting after staining with monoclonal antibodies to CD123, CD11c and a cocktail of lineage markers (CD3, CD14, CD19, and CDF56).\",\n \"Defined numbers of mononuclear cells (start fraction), purified DC1, purified DC2, or mononuclear cells depleted of either DC1 or DC2 by immuno-magnetic affinity column chromatography were added to tissue culture wells in a 96 well plate and cultured with 50 ng/ml GM-CSF and 50 ng/ml IL3 (start, DC2, and DC1 depleted) or 50 ng/ml GM-CSF and 50 ng/ml IL4 (DC1, DC2 depleted).\",\n \"Allo-antigen was prepared by irradiating peripheral blood mononuclear cells from a third party donor to 25 Gy then adding 20,000 cells to each well containing stimulator cells.\",\n \"After 24 and 72 hours of culture in a 5% CO2 atmosphere at 37° C., TNF was added at 10 ng/ml to mature and activate DC progenitors.\",\n \"50,000 autologous naïve T-cells were added to each well and tritiated thymidine incorporation was measured after an additional 96 hours of culture.\",\n \"FIGS.\",\n \"1 and 2 show the results.\",\n \"It is understood that the disclosed invention is not limited to the particular methodology, protocols, and reagents described as these may vary.\",\n \"It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.\",\n \"It must be noted that as used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural reference unless the context clearly dictates otherwise.\",\n \"Thus, for example, reference to “a host cell” includes a plurality of such host cells, reference to “the antibody” is a reference to one or more antibodies and equivalents thereof known to those skilled in the art, and so forth.\",\n \"Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of skill in the art to which the disclosed invention belongs.\",\n \"Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, devices, and materials are as described.\",\n \"Publications cited herein and the material for which they are cited are specifically incorporated by reference.\",\n \"Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.\",\n \"Throughout this application, various publications are referenced.\",\n \"The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.\",\n \"The references disclosed are also individually and specifically incorporated by reference herein for the material contained in them that is discussed in the sentence in which the reference is relied upon.\",\n \"Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein.\",\n \"Such equivalents are intended to be encompassed by the following claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10477012"}}},{"rowIdx":995,"cells":{"text":{"kind":"list like","value":[["Preservation of competent cells at ambient temperatures","The present invention relates to techniques for loading cells with desiccation protectants comprising non-metabolizable and non-reducing carbohydrate analogs.","Specifically, competent bacterial cells can be preserved using the loading techniques provided herein."],["1.A method of preserving competent bacterial cells for storage at ambient temperatures, comprising: incubating said cells with a nonmetabolizable and non-reducing carbohydrate analog, wherein said incubation results in accumulation of said analog in said cells; and drying said cells by foam formation.","2.The method of claim 1, wherein said nonmetabolizable and non-reducing carbohydrate analog is α-methyl-glucoside (MAG) or 2-deoxyglucose (2-DOG).","3.The method of claim 1, wherein said cells are gram negative bacteria.","4.The method of claim 1, wherein said cells are Escherichia coli (E. coli).","5.The method of claim 1, wherein said cells are electrocompetent.","6.The method of claim 1, wherein said cells are chemically competent.","7.The chemically competent cells of claim 6, wherein competence is achieved by mixing said cells with CaCl2 or RbCl.","8.The method of claim 1, further comprising rehydrating the dried cells by contacting said cells with a solution comprising at least one member of the group consisting of carbohydrates, mono-valent cations, divalent cations, organic buffers, and water.","9.The method of claim 8, wherein said carbohydrate is sucrose.","10.The method of claim 8, wherein said cations are calcium or rubidium.","11.The method of claim 1, wherein said carbohydrate analog is administered to said cells at concentrations of 0.1%-50% of total preservation solution.","12.The method of claim 1, wherein said carbohydrate analog is administered to said cells at concentrations of 5%-15% of total preservation solution.","13.The method of claim 1, wherein said logarithmic cells are mid-logarithmic cells.","14.The method of claim 1, wherein said logarithmic cells are late-logarithmic cells.","15.The method of claim 1, wherein said logarithmic cells are harvested at OD550 between 0.1-2.0.16.The method of claim 1, wherein said logarithmic cells are harvested at OD550 between 0.3-1.0.17.The method of claim 1, wherein said logarithmic cells are harvested at OD550 at 0.5.18.The method of claim 1, wherein said incubation is conducted at 0° C.-60° C. 19.The method of claim 1, wherein said incubation is conducted at 20° C.-40° C. 20.The method of claim 1, wherein said incubation is conducted at 30° C.-40° C. 21.The method of claim 1, wherein said incubation is conducted for 0-60 minutes.","22.The method of claim 1, wherein said incubation is conducted for 2.5-60 minutes.","23.The method of claim 1, wherein said incubation is conducted for 30-60 minutes.","24.The method of claim 1, wherein said carbohydrate analogs are actively transported into said logarithmic cells.","25.The method of claim 1, wherein said logarithmic cells are grown in the presence of glucose prior to preservation in order to prime said cells for active transport.","26.The method of claim 25, wherein said logarithmic cells are grown in growth solution comprising 0.001%-50% glucose.","27.The method of claim 25, wherein said logarithmic cells are grown in growth solution comprising 0.050%-5% glucose."],["<SOH> BACKGROUND OF THE INVENTION <EOH>1.Field of the Invention The present invention relates to loading techniques which can be used in conjunction with biological preservation systems.","Specifically, competent cells can be preserved using the loading techniques provided herein.","2.Description of the Related Art Routine procedures in biotechnological laboratories involve daily use of various competent cells for cloning, propagation and preparation of plasmid DNA, construction of genomic libraries, protein expression, and mutagenesis.","Different kinds of competent cells are available commercially, including bacterial, insect, yeast and mammalian cell lines.","Various strains of the gram-negative bacterium Escherichia coli ( E coli ) are one of the most extensively used competent cells.","Depending on the method of transformation that will be applied, competent cells can be prepared as chemically competent (to be transformed by a heat pulse) or as electrocompetent (to be transformed by an electrical pulse).","Chemically competent cells are readily transformable only in the early logarithmic growth stage.","However, early logarithmic cells are extremely sensitive to various stresses, including osmotic and thermal stress, making their preservation difficult.","As an example, preservation survival of <1% in dried logarithmic E. coli cells was recently reported by D. Billi et al.","(Billi, D., D. J. Wright, R. F. Helm, T. Prickett, M. Potts, and J. H. Crowe.","2000.Engineering desiccation tolerance in Escherichia coli.","Appl.","Environ.","Microbiol.","66:1680-1684).","In contrast to chemically competent cells, electrocompetent cells can be transformed in later logarithmic stage and are, therefore, more amenable to preservation.","Regardless of the nature of the competent cells and their final use, the cells must be constantly held at subzero temperatures.","To allow efficient use, competent cells are often prepared in large quantities and then preserved and stored.","Present methodology for the preservation of competent cells includes mixing with different cryoprotectants, for example glycerol, sucrose, dimethly sulfoxide (DMSO), and freezing at −80° C. Because presently available preservation methods include freezing at −80° C., competent cells are routinely subjected to damage by osmotic stress and by exposure to freezing.","Commercially available competent cells are sold individually or as components of various kits developed in order to improve efficiency and accuracy of many routine procedures conducted in research laboratories.","Most of the kit components, except competent cells, can be stored in laboratory refrigerators or at room temperature.","Therefore, the necessity to maintain competent cells at sub-zero temperatures imposes significant burden on distribution of the material and its subsequent storage and use (significant laboratory freezer space is often allocated for the storage of competent cells).","Also, the need for sub-zero temperatures prevents use of competent cells and kits containing the cells in various facilities where freezers are not available.","Therefore, the demand for a technology that will alleviate the need for handling and storing of competent cells at sub-zero temperatures is certainly present."],["<SOH> SUMMARY OF THE INVENTION <EOH>The present invention discloses loading techniques which can be used in conjunction with biological preservation systems.","Specifically, methods for preserving competent cells, such as E. coli, at ambient temperatures are disclosed.","Loading techniques described herein increase desiccation tolerance which allows preservation by foam formation.","In one embodiment of the present invention, a method for preserving competent cells for storage at ambient temperatures is disclosed.","In the methods described therein, cells are incubated with sugar solution and dried by foam formation.","The sugar solution used can be any of a-methyl-glucoside (MAG), 2-deoxyglucose (2-DOG), sucrose, raffinose, or glucose.","In a preferred embodiment, cells contemplated for preservation by the present invention include gram negative bacteria and, specifically, E. coli.","In another embodiment of the present invention, cells may be electrocompetent or chemically competent.","In a preferred embodiment, competence of chemically competent cells is achieved by mixing cells with CaCl 2 or RbCl.","In a further embodiment of the present invention, the preserved cells of the present invention may be rehydrated.","In a preferred embodiment, the rehydration solution may consist of carbohydrates, mono-valent cations, divalent cations, organic buffers, and water.","In an especially preferred embodiment, sucrose is a preferred carbohydrate.","Additionally, calcium and rubidium are preferred cations.","In another aspect of the invention, methods which enhance desiccation tolerance in logarithmic cells are disclosed.","In the methods described therein, cells are incubated with nonmetabolizable and non-reducing carbohydrate analogs.","Preferred analogs of the invention include MAG or 2-DOG.","In a further embodiment of the invention, the carbohydrate analog is administered at concentrations of 0.1%-50% of the total preservation solution.","In an especially preferred embodiment of the invention, the carbohydrate analog is administered at concentrations of 5%-15% of the total preservation solution.","The logarithmic cells of the invention are mid or late logarithmic cells.","Furthermore, in a preferred embodiment, the logarithmic cells are harvested at OD 550 between 0.1-2.0.In another embodiment, the logarithmic cells are harvested at OD 550 between 0.3-1.0.In an especially preferred embodiment, the logarithmic cells are harvested at OD 550 at 0.5.In a further embodiment of the invention, incubation is conducted at 0° C.-60° C. In another embodiment of the invention, incubation is conducted at 20° C.-40° C. In an especially preferred embodiment of the invention, incubation is conducted at 30° C.-40° C. Furthermore, in another aspect of the invention, incubation is conducted for 0-60 minutes.","In another aspect of the invention, incubation is conducted for 2.5-60 minutes.","In an especially preferred embodiment, incubation is conducted for 30-60 minutes.","In another aspect of the invention, the carbohydrate analogs of the invention are actively transported into logarithmic cells.","Additionally, logarithmic cells may be grown in the presence of glucose prior to preservation to prime cells for active transport.","The growth solution of the invention may contain 0.001%-50% glucose.","In an especially preferred embodiment of the invention, the growth solution may contain 0.05%-5% glucose.","According to the present invention, preservation yield of cells dried by foam formation may be increased by incubating cells with sugar solution prior to subjecting the cells to preservation conditions."],["BACKGROUND OF THE INVENTION 1.Field of the Invention The present invention relates to loading techniques which can be used in conjunction with biological preservation systems.","Specifically, competent cells can be preserved using the loading techniques provided herein.","2.Description of the Related Art Routine procedures in biotechnological laboratories involve daily use of various competent cells for cloning, propagation and preparation of plasmid DNA, construction of genomic libraries, protein expression, and mutagenesis.","Different kinds of competent cells are available commercially, including bacterial, insect, yeast and mammalian cell lines.","Various strains of the gram-negative bacterium Escherichia coli (E coli) are one of the most extensively used competent cells.","Depending on the method of transformation that will be applied, competent cells can be prepared as chemically competent (to be transformed by a heat pulse) or as electrocompetent (to be transformed by an electrical pulse).","Chemically competent cells are readily transformable only in the early logarithmic growth stage.","However, early logarithmic cells are extremely sensitive to various stresses, including osmotic and thermal stress, making their preservation difficult.","As an example, preservation survival of <1% in dried logarithmic E. coli cells was recently reported by D. Billi et al.","(Billi, D., D. J. Wright, R. F. Helm, T. Prickett, M. Potts, and J. H. Crowe.","2000.Engineering desiccation tolerance in Escherichia coli.","Appl.","Environ.","Microbiol.","66:1680-1684).","In contrast to chemically competent cells, electrocompetent cells can be transformed in later logarithmic stage and are, therefore, more amenable to preservation.","Regardless of the nature of the competent cells and their final use, the cells must be constantly held at subzero temperatures.","To allow efficient use, competent cells are often prepared in large quantities and then preserved and stored.","Present methodology for the preservation of competent cells includes mixing with different cryoprotectants, for example glycerol, sucrose, dimethly sulfoxide (DMSO), and freezing at −80° C. Because presently available preservation methods include freezing at −80° C., competent cells are routinely subjected to damage by osmotic stress and by exposure to freezing.","Commercially available competent cells are sold individually or as components of various kits developed in order to improve efficiency and accuracy of many routine procedures conducted in research laboratories.","Most of the kit components, except competent cells, can be stored in laboratory refrigerators or at room temperature.","Therefore, the necessity to maintain competent cells at sub-zero temperatures imposes significant burden on distribution of the material and its subsequent storage and use (significant laboratory freezer space is often allocated for the storage of competent cells).","Also, the need for sub-zero temperatures prevents use of competent cells and kits containing the cells in various facilities where freezers are not available.","Therefore, the demand for a technology that will alleviate the need for handling and storing of competent cells at sub-zero temperatures is certainly present.","SUMMARY OF THE INVENTION The present invention discloses loading techniques which can be used in conjunction with biological preservation systems.","Specifically, methods for preserving competent cells, such as E. coli, at ambient temperatures are disclosed.","Loading techniques described herein increase desiccation tolerance which allows preservation by foam formation.","In one embodiment of the present invention, a method for preserving competent cells for storage at ambient temperatures is disclosed.","In the methods described therein, cells are incubated with sugar solution and dried by foam formation.","The sugar solution used can be any of a-methyl-glucoside (MAG), 2-deoxyglucose (2-DOG), sucrose, raffinose, or glucose.","In a preferred embodiment, cells contemplated for preservation by the present invention include gram negative bacteria and, specifically, E. coli.","In another embodiment of the present invention, cells may be electrocompetent or chemically competent.","In a preferred embodiment, competence of chemically competent cells is achieved by mixing cells with CaCl2 or RbCl.","In a further embodiment of the present invention, the preserved cells of the present invention may be rehydrated.","In a preferred embodiment, the rehydration solution may consist of carbohydrates, mono-valent cations, divalent cations, organic buffers, and water.","In an especially preferred embodiment, sucrose is a preferred carbohydrate.","Additionally, calcium and rubidium are preferred cations.","In another aspect of the invention, methods which enhance desiccation tolerance in logarithmic cells are disclosed.","In the methods described therein, cells are incubated with nonmetabolizable and non-reducing carbohydrate analogs.","Preferred analogs of the invention include MAG or 2-DOG.","In a further embodiment of the invention, the carbohydrate analog is administered at concentrations of 0.1%-50% of the total preservation solution.","In an especially preferred embodiment of the invention, the carbohydrate analog is administered at concentrations of 5%-15% of the total preservation solution.","The logarithmic cells of the invention are mid or late logarithmic cells.","Furthermore, in a preferred embodiment, the logarithmic cells are harvested at OD550 between 0.1-2.0.In another embodiment, the logarithmic cells are harvested at OD550 between 0.3-1.0.In an especially preferred embodiment, the logarithmic cells are harvested at OD550 at 0.5.In a further embodiment of the invention, incubation is conducted at 0° C.-60° C. In another embodiment of the invention, incubation is conducted at 20° C.-40° C. In an especially preferred embodiment of the invention, incubation is conducted at 30° C.-40° C. Furthermore, in another aspect of the invention, incubation is conducted for 0-60 minutes.","In another aspect of the invention, incubation is conducted for 2.5-60 minutes.","In an especially preferred embodiment, incubation is conducted for 30-60 minutes.","In another aspect of the invention, the carbohydrate analogs of the invention are actively transported into logarithmic cells.","Additionally, logarithmic cells may be grown in the presence of glucose prior to preservation to prime cells for active transport.","The growth solution of the invention may contain 0.001%-50% glucose.","In an especially preferred embodiment of the invention, the growth solution may contain 0.05%-5% glucose.","According to the present invention, preservation yield of cells dried by foam formation may be increased by incubating cells with sugar solution prior to subjecting the cells to preservation conditions.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is a bar chart showing preservation survival of electrocompetent E. coli cells preserved by foam formation.","FIG.","2 is a bar chart showing electroporation survival of electrocompetent E. coli cells preserved by foam formation.","FIG.","3 is a bar chart showing electroporation efficiency of electrocompetent E. coli cells preserved by foam formation.","FIG.","4 is a line graph showing the effect of increasing concentration of glucose analogs during in vitro loading on preservation survival of logarithmic E. coli.","FIG.","5 is a line graph showing the effect of loading temperature on preservation survival of E. coli.","FIG.","6 is a line graph showing the effect of loading temperature on stability of the preserved E. coli.","FIG.","7 is a line graph showing the effect of loading time on preservation survival and stability of E. coli cells loaded with 10% MAG at 37° C. FIG.","8 is a line graph showing the effect of growth stage on the preservation survival of E. coli loaded with 10% MAG.","DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT The present invention discloses loading techniques which can be used in conjunction with biological preservation systems.","Specifically, the invention relates to methods of using loading techniques to increase desiccation tolerance that will allow preservation by foam formation.","In a preferred embodiment, the methods described herein may be used to preserve E. coli cells.","In order to preserve competent cells under conditions less damaging then freezing and to allow their subsequent handling at temperatures above sub-zero, we used the foam formation process.","Foam formation technology represents an advanced method for preservation by drying, where the sample temperature is always kept above freezing (U.S. Pat.","No.","5,766,520).","Therefore, the possibility for cell damage by formation of ice-crystals, which is the most common damage during preservation by freezing, is eliminated.","However, because drying itself imposes significant osmotic stress on logarithmic cells, cellular desiccation tolerance should be optimized in order to take the full advantage of the foam formation preservation process.","We demonstrated that incubation of the cells with an appropriate sugar solution prior to preservation (“loading”), can significantly increase preservation yield of cells dried by foam formation.","Herein we describe a method to preserve E coli cells at ambient temperature by the foam formation process and demonstrate that the preserved cells are transformable at efficiency comparable to that of the fresh cells.","Preservation of Electrocompetent Cells Preservation of electrocompetent cells in the various working examples included infra were conducted as follows, unless otherwise stated: (1) Preparation of fresh electrocompetent cells: Escherichia coli ATCC strain MM294 cells were grown under standard conditions in L-broth (growth medium for E. coli), at 37° C., with moderate aeration (shaking at 250 rpm).","An isolated colony from a fresh streak was inoculated into 5 ml of L-broth and incubated overnight (preculture).","The following morning, the preculture was subcultured (1% inoculum) in 100 ml of L-broth+glucose (0.05%-0.5%).","The cells were incubated until OD550=0.5 (subculture).","At this point, the cells were transferred into 250 ml of L-broth+glucose (0.05%-0.5%) and incubation continued until OD550=0.8.Two flasks with 250 ml of cells were prepared.","When the culture OD550 reached 0.8 (cell density approx.","5×108 cfu/ml), the cells were harvested by centrifugation at 2,500 rpm (at room temperature) for 10 minutes.","The cells (2×200 ml samples) were washed once with an equal volume of PBS buffer or twice with an equal volume of 20% ice-cold sucrose and centrifuged again as described above.","The cells were concentrated 100-fold in PBS buffer or in 20% sucrose to cell density of approx.","2×1010 cfu/ml.","Bacterial counts were determined by diluting cells in PBS buffer and spread plating appropriate dilutions in duplicate on L-agar.","The plates were incubated at 37° C. overnight and colonies were counted the following day.","(2) Electroporation of fresh E. coli electrocompetent cells: Two aliquots of the concentrated cells (50 μl each) were transferred into pre-chilled Eppendorf tubes (1.5 ml) and 2 μl of pUC18 plasmid DNA (approx.","300 ng/μl) were added to the tubes.","The [cells+DNA] mixtures were incubated on ice for 5 minutes and transferred into pre-chilled cuvettes for electroporation.","The cells were electroporated by using a BTX T-100 electroporator, at 625 or at 800 volts (V) in BTX cuvettes (0.1 cm gap, P/N 610 Disposable Cuvettes) with a time constant “r” equal 5 msec.","Under the pulse conditions described above, the cells were subjected to an electrical field “E” equal to 6.25 kV/cm or 8.0 kV/cm.","Immediately after the pulse, the cells were transferred into small glass tubes (10×100 mm) with 0.5 ml of “outgrowth medium” (L-broth+20% sucrose+10 mM CaCl2).","The electroporated bacteria were incubated at 37° C. with moderate shaking (150 rpm) for 60 minutes.","When the 60 minutes period elapsed, the cells were diluted in “outgrowth medium” and duplicate samples were spread plated on L-agar plates with no antibiotic (to determine electroporation survival) and on L-agar plates with 50 μg/ml or 100 μg/ml of ampicillin (selective marker on pUC18 plasmid; to determine electroporation efficiency).","The plates were incubated overnight at 37° C. and the colonies were counted the next day.","(3) Loading of fresh electrocompetent E. coli cells before preservation: When applicable, one aliquot of the cell concentrate (3.2 ml of cells in PBS buffer, approx.","2×1010 cfu/ml) was mixed with 0.8 ml of 50% methyl-glucoside (MAG; 10% final concentration) and incubated at 37° C. for 30 minutes (in vitro loading).","A second aliquot (3.2 ml of the cell concentrate) was mixed with 0.8 ml of PBS buffer and incubated on ice for 30 minutes (control, unloaded cells).","Similar to loading with MAG, the cells were loaded with 2-deoxyglucose (2-DOG) as described in Example 5 infra.","(4) Preservation of electrocompetent E. coli cells: The cells (resuspended in PBS buffer or on 20% sucrose) were mixed with an equal mass of preservation solution consisting of 45% 2:1 sucrose:raffinose or 45% 4:1 sucrose:MAG on ice.","Altematively, the cell pellets were resuspended directly in 22.5% sucrose:raffinose or 22.5% 4:1 sucrose:MAG.","The 0.5 g aliquots were distributed in 5 ml glass serum vials or 0.2 g aliquots were distributed in 1.2 ml glass serum vials.","To determine the survival in preservation solution before drying, cell-containing mixtures were diluted in PBS buffer containing 20% sucrose and appropriate dilutions were spread plated in duplicate on L-agar plates.","The samples were dried by foam formation.","The mixtures were foamed at 0° C. and kept at that temperature until foam formation occurred.","After foam was well formed, shelf temperature was increased to 20° C. and drying continued overnight (at least 12 hours).","When applicable, electrocompetent cells were frozen in the following manner.","The cell suspension (100 μl) was mixed with 10% DMSO (10 μl) and slowly frozen at −80° C. (5) Assaying the preserved E. coli cells: Dried samples were rehydrated at room temperature.","1 ml of rehydration solution (20% sucrose) was added to the 5 ml vials and 0.2 ml was added to the 1.2 ml vials.","For the determination of preservation survival, rehydrated cells were diluted and spread plated in duplicate on L-agar plates.","Plates were incubated at 37° C. overnight and colonies were counted the following day.","(6) Electroporation of dried (preserved) E. coli cells: One or two 50 μl aliquots were removed from each rehydrated vial and transferred to pre-chilled Eppendorf tubes.","The cells were mixed with pUC18 plasmid DNA (300 ng/μl) and electroporated using the conditions described previously for the electroporation of fresh electrocompetent cells.","Electroporation survival was calculated as the number of viable cells recovered after electrical pulse per total number of electroportated cells.","Electroporation efficiency was calculated as the number of transformants (cells which received pUC18 plasmid DNA) per number of cells recovered after electroporation (electroporation survival).","When applicable, the electroporation efficiency was also expressed as number of transformants per 1 μg of pUC18 DNA (Cfu/μg).","Preservation of Chemically Competent Cells Preservation of chemically competent cells in the various working examples included infra was conducted as follows, unless otherwise stated: (1) Preparation of chemically competent cells: The cells were grown as described for preparation of electrocompetent cells, except that the culture was harvested when OD550 reached 0.5.The cells were chilled on ice and centrifuged as described for electrocompetent cells.","The supernatant was removed and the cells were and re-suspend in 30 ml of ice-cold Buffer A [30 mM KOAc, 50 mM MnCl2, 100 mM RbCl, 10 mM CaCl2, 10% sucrose), pH=5.8.The cells were incubated on ice for 120 minutes.","When the 120 minutes period elapsed, the cells were centrifuged for 5 minutes at 2,500 rpm in a pre-chilled centrifuge.","The supernatant was removed and the cells resuspended in 4 ml of ice-cold Buffer B [10 mM Na MOPS (pH=7.0), 75 mM CaCl2, 10 mM RbCl, 10% sucrose].","The cells were kept on ice at all times.","(2) Preservation of chemically competent E. coli cells: The cells were transferred into ice-cold 15 ml Falcon tubes.","3.5 ml of cells were mixed with 3.5 g of ice-cold preservation solution consisting of 40% 3:1 sucrose:MAG in PBS buffer.","The 2×0.1 ml of preservation mixture were removed, serially diluted with PBS buffer and plated in duplicate on agar plates with no antibiotic (“preservation mixture controls”).","The 0.5 g aliquots of preservation mixture were distributed into pre-chilled 5 ml sterile glass vials.","The mixtures were preserved by foam formation as described for drying of electrocompetent cells.","(3) Assaying of the preserved chemically competent E. coli cells: Two vials of the dried cells were transferred on ice and re-hydrated with 0.5 ml of Buffer B.","For the determination of preservation survival, rehydrated cells were diluted in buffer B and spread plated in duplicate on L-agar plates.","Plates were incubated at 37° C. overnight and colonies were counted the following day.","The vials were stored in a refrigerator for future assays.","(4) Transformation of chemically competent E. coli cells by “heat shock”: The fresh and the preserved cells were transformed under identical conditions, except that in order to keep the same cell density during the transformation, 0.2 ml of the preserved cells and 0.1 ml of the fresh cells were used.","The cells were transformed in the following manner.","0.1 ml of the fresh cells or 0.2 ml of re-hydrated content from each vial were transferred into pre-chilled Eppendorf tubes.","The pUC18 plasmid DNA (5 μl, approx.","500 ng) was added into each Eppendorf tube with cells.","The cells and DNA were mixed gently and the mixtures were incubated on ice for 120 minutes.","When the 120 minutes period elapsed, the cells were transferred for 90 seconds at 42° C. (“heat shock”).","The cells were returned on ice for 2 minutes and then transferred into 5 ml glass tubes containing 5 volumes of L-broth pre-warmed at room temperature.","The cells were shaken gently at 37° C. for 60 minutes (150 rpm).","Following the 60 minutes incubation at 37° C., 0.1 ml aliquots of the cells were removed from the tubes, diluted with L-broth medium and spread-plated in duplicate on L-agar plates with 50 μg/ml of ampiclin (Ap50).","The plates were incubated aerobically at 37° C. overnight.","The following working examples illustrate preservation of electrocompetent and chemically competent cells by the use of loading in accordance with the methods of the present invention: EXAMPLE 1 Electrocompetent E. coli MM294 cells were prepared as described previously, except for as follows: In this experiment, the cells were washed twice in ice-cold 20% sucrose and concentrated 100-fold after the second wash. Additionally, four samples were prepared (A, B, C, and D).","The cell concentrates were diluted 1:1 with the preservation solution consisting of 45% 2:1 sucrose:raffinose (sample A) or resuspended directly in 22.5% 2:1 sucrose:raffinose (samples B and C).","Samples A and B were preserved in 0.5 ml vials (0.5 g fill per vial).","Sample C was preserved in 1.2 ml vials (0.2 g fill per vial).","Additional solution consisting of 22.5% 4:1 sucrose:MAG was prepared and evaluated for preservation of the cells (sample D).","The cell concentrate (“D”) was resuspended directly in 22.5% 4:1 sucrose:MAG and preserved in 1.2 ml vials (0.2 g fill per vial).","Dried cells of all samples were rehydrated in 20% sucrose and assayed for stability (1 ml was added to the cells in 5 ml vials, 0.2 ml to the cells in 1.2 ml vials).","The preserved cells of sample B were rehydrated with nano-pure water (0% sucrose), 10%, and 20% sucrose and electroporated.","Electroporation survival and efficiencies in samples rehydrated with solutions containing different amounts of sucrose were determined.","Finally, fresh and dried cells were electroporated at 800V.","Electroporation survival and efficiencies were determined and compared.","Preservation yield, electroporation survival and efficiencies obtained with the cells in four different samples are shown in Table 1.TABLE 1 Sample Preservation Electroporation Electroporation Description Survival (%) Survival (%) Efficiency (10−3%) Fresh Cells ND 95.8 +/− 17.2 0.50 +/− 0.06 Foam Formation Preserved (Day 0) A 36.8 +/− 2.7 36.6 +/− 3.9 8.1 +/− 0.8 B 21.5 +/− 5.6 70.2 +/− 12.4 2.2 +/− 0.5 C 18.5 +/− 3.2 43.0 +/− 8.1 2.7 +/− 0.2 D 18.7 +/− 2.9 63.9 +/− 11.4 3.7 +/− 0.2 Foam Formation Preserved (Day 11 at 4° C.) A 39.8 +/− 4.5 15.1 +/− 1.7 19.8 +/− 2.5 B 30.2 +/− 3.2 19.9 +/− 7.7 3.3 +/− 1.7 C 22.7 +/− 5.8 19.1 +/− 2.4 3.7 +/− 0.9 D 32.2 +/− 7.8 13.8 +/− 1.0 10.4 +/− 0.9 Foam Formation Preserved (Day 11 at RT°) A 36.2 +/− 6.4 19.5 +/− 1.5 21.1 +/− 5.1 B 25.9 +/− 1.5 25.5 +/− 5.9 4.7 +/− 1.1 C 23.6 +/− 8.6 10.1 +/− 5.6 8.2 +/− 1.6 D 27.6 +/− 6.5 15.0 +/− 6.6 8.2 +/− 1.9 Foam Formation Preserved (Day 20 at 4° C.) A 38.9 +/− 10.9 30.6 +/− 2.0 17.2 +/− 5.5 B N/D N/D N/D C N/D N/D N/D D N/D N/D N/D Foam Formation Preserved (Day 20 at RT°) A 30.3 +/− 2.3 33.9 +/− 2.9 22.8 +/− 8.2 B 17.8 +/− 5.3 22.3 +/− 3.5 5.6 +/− 3.1 C 27.4 +/− 10.4 18.2 +/− 3.6 5.9 +/− 0.8 D 21.7 +/− 0.5 29.9 +/− 6.7 5.1 +/− 0.8 The cell concentrate diluted 1:1 in the preservation solution (A) preserved at a higher yield compared to the concentrate (B) resuspended directly in preservation solution (36.8% versus 21.5%).","There was no significant difference in the preservation yield or stability of the preserved samples with respect to the preservation solution used (18.5% in sample C and 18.7% in sample D at Day 0).","Regardless of the concentration of the cells in a sample or a vial size, the preserved cells were stable for 20 days at RT and at 4° C. Comparison of the electroporation parameters in preserved and fresh cells revealed that electroporation survival in the preserved cells was lower than in the fresh cells.","In contrast, electroporation efficiency in the preserved cells was higher.","The preserved cells routinely electroporated at the efficiency 3 to 10 fold higher than the fresh cells.","With respect to electroporation parameters in the preserved cells, electroporation survival at Day 0 was higher when the cells were more concentrated (sample B versus sample A).","When the cells were preserved in small aliquots (0.2 g in 1.2 ml vial), electroporation survival was higher in the cells preserved in sucrose:MAG (D) compared to the cells preserved in sucrose:raffinose (C).","Differences in electroporation survival between different samples were smaller at Day 20 than at Day 0.At Day 0, electroporation efficiency was somewhat higher in the preserved cells in sample A than in sample B.","Similar to electroporation survival, the electroporation efficiency was slightly higher at Day 0 in the cells preserved in sucrose:MAG compared to the cells preserved in sucrose:raffinose.","At all times (Day 0-Day 20), transformation efficiency was significantly higher in the preserved cells in sample A compared to the cells in other samples.","The effect of the sucrose concentration in the rehydration medium on preservation survival and electroporation parameters was evaluated.","Relevant data are presented in Table 2.TABLE 2 Preserved cells of sample B rehydrated with different amounts of sucrose Preservation Electroporation Electroporation Sample Description Survival (%) Survival (%) Efficiency (10−3%) 0% sucrose 14.8 +/− 2.9 37.3 +/− 6.3 45.6 +/− 1.5 10% sucrose 32.4 +/− 8.0 35.4 +/− 4.0 10.7 +/− 1.4 20% sucrose 23.4 +/− 2.8 74.5 +/− 8.2 0.9 +/− 0.3 According to data presented in Table 2, the cells rehydrated without sucrose had lower preservation yield than the samples rehydrated with 10% or 20% sucrose (14.8% versus 23-32%).","Similar to the preservation survival, the electroporation survival was higher in the preserved cells rehydrated in the solution containing the higher concentration of sucrose (74.5% in 20% sucrose versus 37.3% with no sucrose).","Increase in the amount of sucrose in the rehydration solution had an inhibitory effect on electroporation efficiency.","The efficiency in the cells rehydrated without sucrose was 45.6% compared to 0.9% in the cells rehydrated with 20% sucrose (Table 2).","Based on the data from this example, the following conclusions were reached: 1.Electrocompetent E. coli cells could be successfully preserved by foam formation.","2.There was no difference in preservation survival when the cells were preserved in small aliquots (0.2 g in 1.2 ml vial compared to 0.5 g in 0.5 ml vial).","3.Preservation survival and stability were slightly better in diluted cells (A versus B).","4.There was no significant difference in preservation survival when the cells were preserved in sucrose:raffinose or in sucrose:MAG.","5.Electroporation survival in the preserved cells was lower than in the fresh cells.","6.Electroporation efficiency in the preserved cells was routinely higher than in the fresh cells.","Efficiency was the highest in the cells which were diluted 1:1 with preservation solution.","7.Increase in the amount of sucrose in rehydration medium enhanced preservation and electroporation survival, but was inhibitory with respect to the electroporation efficiency.","EXAMPLE 2 To enhance bacterial desiccation tolerance, the cells were loaded with 10% MAG.","The yield of the preserved electrocompetent E. coli cells, electroporation survival and electroporation efficiency of fresh and preserved cells (loaded and unloaded control) were evaluated and compared (Table 3, FIG.","1, FIG.","2, and FIG.","3).","TABLE 3 Electroporation Electroporation Preservation Yield Survival Efficiency Cells Cfu/ml % Cfu/ml % Cfu/ml 10−3% Fresh 2.0 × 1010 N/A 8.6 × 109 43.5 2.1 × 105 2.4 Preserved (Dried) Control 3.8 × 109 19.3 6.3 × 108 16.5 1.6 × 105 25.4 (Treatment A) Loaded 1.1 × 1010 53.5 3.0 × 109 28.6 1.7 × 105 5.6 (Treatment B) N/A, not applicable.","The data represents the average of three independent experiments.","The preservation yield of the loaded dried electrocompetent cells was significantly higher than the yield in unloaded controls (53.5% versus 19.3%, Table 3).","Therefore, loading with MAG ameliorated desiccation damage during foam formation resulting in increased preservation survival.","Electroporation survival of dried cells was lower than in fresh cells (Table 3).","In dried loaded cells, electroporation survival was higher than in dried control (28.6% versus 16.5%, Table 3).","Electroporation efficiency of the preserved loaded cells was somewhat higher than in the fresh cells (5.6×10−3% versus 2.4×10−3%, Table 3).","The following conclusions were made based on the data from this example: 1.Loading of the cells with 10% MAG significantly increased preservation survival.","2.Electroporation survival in the preserved cells was lower than in the fresh cells.","3.Similar to preservation survival, loading of the cells with 10% MAG significantly increased electroporation survival.","4.Electroporation efficiency (cfu/ml) in the preserved cells was comparable or slightly higher compared to the fresh cells.","EXAMPLE 3 In this example, commercial electrocompetent E. coli cells, in the form of a bacterial pellet, were preserved by the foam formation process.","The cells were concentrated 100-fold and preserved as described previously.","To increase desiccation tolerance and enhance preservation survival.","One aliquot of the cells were loaded in vitro with 10% MAG.","The preserved cells were stored at 4° C. In addition to preservation by foam formation, the cells were preserved by slow freezing (with 10% DMSO) at −80° C. Preservation yields and stability in commercial electrocompetent E. coli cells preserved by foam formation and by freezing are presented in Table 4.TABLE 4 Preservation yield** Stability (12 days) Cells Cfu/ml % Cfu/ml % Fresh 1.4 × 1010 +/− 2.8 × 109 100 +/− 19.9 N/A N/A Frozen* 1.1 × 1010 +/− 2.1 × 109 75.6 +/− 14.8 1.2 × 1010 +/− 5.4 × 108 82.7 +/− 3.9 Preserved (Control)* 6.6 × 109 +/− 4.5 × 108 47.7 +/− 3.2 4.9 × 109 +/− 1.5 × 109 35.8 +/− 10.9 Preserved (Loaded) 8.8 × 109 +/− 3.9 × 108 63.6 +/− 2.8 9.5 × 109 +/− 1.1 × 108 68.5 +/− 0.8 *The control cells were also assayed for stability after 67 days at 4° C. Some loss in viability was observed (20 +/− 1% survival, 3 × 109 +/− 2.0 × 108 cfu/ml).","**Frozen cells were incubated 72 hours at −80° C.; Foam formation-preserved cells were dried overnight.","N/A, not applicable.","The control (unloaded) commercial cells dried by foam formation were preserved at a 47.7% yield.","The cells were relatively stable at 4° C. (35.8% viability after 12 days and 20% viability after 67 days storage).","Commercial cells loaded in vitro with 10% MAG and preserved by foam formation had a yield of 63.6%.","These cells were completely stable at 4° C. (68.5% yield after 12 days).","The frozen cells were also stable after 12 days of storage at −80° C. Preserved commercial cells (vitrified and frozen) were electroporated as described previously.","Electroporation survival and electroporation efficiency in the preserved cells were determined and compared to the same parameters in the fresh cells (Table 5).","TABLE 5 *Preserved *Preserved Cells Fresh (Control) (Loaded) Electroporation Survival (Cfu/ml) 6.40 × 109 +/− 1.3 × 109 7.9 × 108 +/− 1.2 × 108 3.2 × 109 +/− 4.2 × 108 (%) 46.2 +/− 9.4 12.2 +/− 1.7 35.7 +/− 4.7 Electroporation Efficiency (Cfu/ml) 1.5 × 105 +/− 4.0 × 104 7.0 × 104 +/− 5.1 × 104 1.9 × 105 +/− 1.7 × 105 (10−3%) 2.4 +/− 0.6 8.8 +/− 6.4 6.1 +/− 5.5 (Cfu/□g DNA) 4.6 × 104 +/− 1.2 × 104 4.3 × 103 +/− 3.1 × 103 1.6 × 104 +/− 1.8 × 102 *Cell density in the preserved cells was {fraction (1/10)} of that in fresh concentrates.","These cells were transformed immediately after drying and they were not concentrated before electroporation.","Electroporation survival in the preserved cells was lower than in the fresh cells (12.2% or 35.7% versus 46.2%).","In contrast, electroporation efficiency in the preserved cells was higher than in the fresh cells (6.1% or 8.8% versus 2.4%).","Electroporation survival in preserved control cells was lower than in preserved loaded cells.","Electroporation efficiencies were comparable in unloaded and loaded preserved cells (6.1% versus 8.8%).","Based on the data from this example, the following conclusions were made: 1.Commercial electrocompetent E. coli cells were successfully preserved by the foam formation process.","2.Loading with MAG increased stability of the preserved cells at 4° C. 3.Electroporation survival was higher in fresh cells than in the preserved cells.","4.In preserved cells, electroporation survival was higher in loaded cells compared to unloaded control.","5.Electroporation efficiency was higher in the preserved cells compared to that in the fresh cells.","6.There was no significant difference in electroporation efficiency in MAG-loaded preserved cells versus unloaded preserved controls (MAG was slightly inhibitory).","EXAMPLE 4 Chemically competent E. coli MM294 cells were prepared and preserved by foam formation as described previously.","Preservation yield and stability of the preserved cells are described in Table 6.TABLE 6 Preservation Yield (Day 0) Stability (290 Days at 4° C.) Cells Cfu/ml % Cfu/ml % Fresh 1.6 × 1010 +/− 6.8 × 109 N/A N/A N/A Preserved 5.1 × 107 +/− 4.0 × 107 0.3 +/− 0.3 1.1 × 107 +/− 4.5 × 106 0.07 +/− 0.03 Chemically competent E. coli MM294 cells were preserved at low yield (0.3%).","The cells were harvested in early logarithmic stage and were not loaded with MAG or any other sugar derivative to enhance bacterial desiccation tolerance and the subsequent preservation survival.","The preserved cells were relatively stable at 4° C. After 290 days of storage, the observed loss in viability was less than 1 Log (Table 6).","Transformation efficiencies of the fresh and the preserved chemically competent cells are presented in Table 7.TABLE 7 Transformation Efficiency Day 0 290 Days at 4° C. Cells Cfu/ml 10−3% Cfu/ml 10−3% Fresh 9.7 × 105 +/− 1.9 × 105 6.1 +/− 1.1 N/A N/A Preserved 6.7 × 102 +/− 2.4 × 102 1.3 +/− 0.5 1.6 × 102 +/− 5.0 × 101 1.5 +/0.5 Transformation efficiency in the preserved cells was somewhat lower compared to the efficiency of the fresh cells (Table 7).","Preservation of the cells by foam formation did not compromise bacterial competence in any significant fashion.","Transformation efficiency of the preserved cells stored for 290 days at 4° C. was comparable to the efficiency immediately after drying.","Based on the data of this example, the following conclusions were reached: 1.Chemically competent E. coli cells could be successfully preserved by foam formation.","2.Preservation yield was low because the cells were harvested in early logarithmic phase when they likely had no internal desiccation protectants.","3.The cells remained relatively stable upon prolonged storage at 4° C. 4.The preserved cells remained competent after preservation.","5.The competence function was not compromised during storage at 4° C. EXAMPLE 5 Accumulation of MAG in vitro enhances preservation survival of logarithmic E. coli cells.","Preparation of electrocompetent cells routinely requires harvesting bacterial cultures in late logarithmic growth stage.","In contrast, the cells must be harvested in an early or medium logarithmic growth stage for preparation of chemically competent cells.","Late logarithmic cells are more tolerant to desiccation than early logarithmic cells.","We found that desiccation tolerance of logarithmic E. coli could be significantly enhanced by loading the cells with MAG.","We also found that efficiency of loading could be influenced by the following parameters: 1.Choice of carbohydrate 2.Loading temperature 3.Loading time 4.Growth stage of the cells Accumulation of non-metabolizable glucose analogs enhances preservation of logarithmic E. coli cells.","Bacterial culture was grown until OD550=0.5 was reached (mid-log growth stage).","The cell concentrates were prepared as described previously.","Control cells, which were not loaded, were incubated on ice for 30 minutes.","Cell concentrates were incubated with different amounts (0.15-1%, 5%, 10% and 15%) of glucose and glucose nonmetabolizable analogs at 37° C. for 30 minutes (loading).","These mixtures were preserved as described previously.","Mid-logarithmic cells loaded with 0.1%/-1% and 5% of MAG survived drying at 9%, 12% and at 24%, respectively.","When cells were loaded with 0.1-1%, 5%, 10% and 15% of 2-DOG, bacteria survived drying at 9-12%, 20%, 16%, and 20%, respectively (FIG.","4).","Loading with more than 5% of either MAG or 2-DOG in vitro did not improve drying survival compared to the survival obtained when 5% of either compound was used.","Control cells, which were not loaded, survived drying at <1%.","In contrast to its nonmetabolizable analogs MAG or 2-DOG, glucose had no protective effect on preservation survival of logarithmic cells.","Mid-logarithmic cells loaded in vitro with up to 15% of glucose survived drying at 1-3% (FIG.","4).","There was no significant difference in survival when different amounts of glucose were used for loading.","Because glucose was used at concentrations significantly higher than physiological, likely only a portion of the sugar was metabolized.","Remaining glucose had no immediate damaging effect on preservation survival.","However, because of its high reducing activity, glucose could be detrimental to cells due to a non-enzymatic browning, including the Mallard's reaction.","Based on the data of this example, the following conclusions were made: 1.Incubation of mid-logarithmic E. coli cells with 5% of MAG or 2-DOG prior to drying enhanced preservation survival of bacteria.","2.Incubation with more than 5% of nonmetabolizable sugar analogs had no additional effect on preservation survival.","3.Accumulation of glucose in the cells had no protective effect against desiccation stress during preservation.","EXAMPLE 6 Accumulation of MAG in the cells prior to drying enhances preservation of logarithmic E. coli cells.","Two cell concentrates (harvested at mid-log growth stage) were prepared as described previously.","To induce uptake of MAG, 0.5% glucose was added to the growth medium (“induced culture”).","Both concentrates were incubated with 10% MAG at 37° C. for 30 minutes (loading).","When loading was completed, one concentrate was diluted 100-fold in PBS buffer and incubated at 37° C. for an additional 30 minutes to induce expulsion of the pre-accumulated MAG.","Both concentrates were preserved as described previously.","Cell density in the concentrates was 4.7×108 cfu/ml.","Preservation yield in loaded concentrate, which was not subsequently diluted, was 57% (2.7×108 cfu/ml).","In contrast, in the concentrate, which was diluted to remove MAG pre-accumulated in the cells, preservation yield was only 3.8% (1.8×107 cfu/ml).","Cells loaded with MAG were stable at room temperature for extended periods of time (i.e.","stability after 12 days was 52%, 2.5×108 cfu/ml).","Cells, which were depleted from pre-accumulated MAG by expulsion, and preserved at low yield, remained at 2.4% (1.1×107 cfu/ml).","Based on the data of this example, the following conclusions were made: 1.Accumulation of non-metabolizable carbohydrate analogs, such as MAG, prior to drying enhances preservation survival of mid-log cells.","2.Removal of the accumulated sugar by expulsion results in sensitivity to desiccation and low preservation yield.","EXAMPLE 7 Mid-logarithmic culture was prepared as described previously.","0.5% glucose was added to the growth medium (“induced cells”).","The culture was concentrated 10-fold and the concentrates were incubated for 30 minutes with 10% MAG at different temperatures (0, 20, 40, and 60° C.).","The mixtures were preserved as described previously.","Cell densities (cfu/ml) in mid-logarithmic E. coli culture were 1.6×108 cfu/ml.","Cell density in the concentrate was 1.9×109 cfu/ml.","Preservation yield and stability of E. coli cells incubated with 10% MAG at different temperatures are shown in Table 8 and in FIGS.","5 and 6.TABLE 8 Preservation Yield Stability (9 Days at RT°) Stability (16 Days at RT°) Sample % % % Description Cfu/ml Survival Cfu/ml Survival Cfu/ml Survival Loaded at 1.6 × 108 +/− 8.2 +/− 1.4 9.4 × 107 +/− 5.0 +/− 0.6 9.9 × 107 +/− 5.2 +/− 1.5 0° C. 2.7 × 107 1.2 × 107 2.8 × 107 Loaded at 2.7 × 108 +/− 14.1 +/− 2.6 1.6 × 108 +/− 8.7 +/− 0.6 1.4 × 108 +/− 7.3 +/− 1.7 20° C. 4.9 × 107 6.7 × 107 3.2 × 107 Loaded at 4.4 × 108 +/− 23.2 +/− 3.5 5.3 × 108 +/− 28.1 +/− 3.5 3.9 × 108 +/− 21.0 +/− 1.9 40° C. 6.6 × 107 6.6 × 107 3.7 × 107 Loaded at 1.4 × 104 +/− 0 1.2 × 104 +/− 0 1.3 × 104 +/− 0 60° C. 1.4 × 10−3 2.3 × 103 1.9 × 103 Logarithmic cells incubated with 10% MAG at 20° C. or at 40° C. preserved at higher yields and were more stable after 16 days of storage at room temperature compared to the cells incubated with MAG at 0° C. and at 60° C. (FIGS.","5 and 6).","Cells incubated with MAG at 40° C. preserved at higher yield than cells incubated with MAG at 20° C. and remained more stable during storage.","Incubation at 60° C. resulted in severe loss in bacterial viability.","Based on the data from this example, the following conclusions were reached: 1.Efficiency of loading of MAG into logarithmic E. coli cells is highly dependent on the incubation temperature.","2.Loading of mid-log E. coli cells with MAG was the most effective when the cells were incubated at 20°-40° C., preferentially closer to 40° C. EXAMPLE 8 Effect of incubation time on accumulation of MAG in logarithmic E. coli cells.","Cell concentrates were prepared as described previously (0.5% glucose was added to the growth medium) and incubated with 10% MAG at 37° C. for 0, 2.5, 5, 15, 30, and 60 minutes.","The mixtures were preserved as described previously.","Preservation yield and stability of mid-logarithmic E. coli cells incubated with 10% MAG for 0-60 minutes are shown in Table 9 and on FIG.","7.TABLE 9 Preservation Yield Stability (7 days at RT) Stability (17 days at RT) Sample % % % Description Cfu/ml Survival Cfu/ml Survival Cfu/ml Survival Incubated 5.3 × 107 +/− 1.7 × 107 2.6 +/− 0.8 2.4 × 107 +/− 1.2 +/− 0.7 3.6 × 107 +/− 1.8 +/− 0.5 for 0 min 1.4 × 107 1.1 × 107 Incubated 1.2 × 108 +/− 2.1 × 107 5.8 +/− 1.1 9.6 × 107 +/− 4.9 +/− 0.8 5.9 × 107 +/− 2.9 +/− 0.6 for 2.5 min 1.5 × 107 1.2 × 107 Incubated 2.0 × 108 +/− 1.1 × 107 9.9 +/− 0.6 1.4 × 108 +/− 6.8 +/− 1.2 1.9 × 108 +/− 9.5 +/− 2.7 for 5 min 2.4 × 108 5.4 × 107 Incubated 3.4 × 108 +/− 5.6 × 107 16.8 +/− 2.8 2.1 × 108 +/− 10.4 +/− 0.4 2.4 × 108 +/− 11.8 +/− 2.6 for 15 min 8.3 × 106 5.2 × 107 Incubated 4.0 × 108 +/− 4.4 × 107 19.9 +/− 2.2 2.6 × 108 +/− 13.0 +/− 2.7 3.4 × 108 +/− 16.8 +/− 2.6 for 30 min 5.4 × 107 5.2 × 107 Incubated 4.4 × 108 +/− 6.1 × 107 22.2 +/− 3.0 3.3 × 108 +/− 16.3 +/− 1.9 3.9 × 108 +/− 19.3 +/− 3.9 for 60 min 3.7 × 107 7.7 × 107 Logarithmic cells incubated with MAG for 2.5-60 minutes preserved at higher yields than cells incubated with MAG for 0-2.5 minutes.","Preservation survival of mid-logarithmic E. coli cells was directly proportional to the time of incubation with MAG (FIG.","7).","After 17 days of storage at RT, the cells incubated with MAG for at least 2.5 minutes were more stable than the cells incubated with MAG for less than 2.5 minutes.","The following conclusions were reached, based on the data of this example: 1.Incubation with MAG for at least 2.5 minutes enhances preservation survival of mid-log E. coli cells.","Loading for at least 5 minutes enhanced stability of the preserved cells.","2.Preservation yield and stability of the preserved cells are directly proportional to the length of incubation with MAG.","EXAMPLE 9 Effect of growth stage on accumulation of MAG in E. coli cells.","E. coli cultures were grown in L-broth with 0.5% glucose as described previously and harvested at OD550=0.1 (early-log), 0.5 (mid-log), 1.0 (late-log) and 2.0 (stationary cells).","The cells in each culture were concentrated and split in two aliquots.","One aliquot of each concentrate was incubated with 10% MAG at 37° C. for 30 minutes (“loaded”).","The second aliquot was not loaded (controls).","Unloaded controls and loaded cells were preserved as described previously.","Cell densities at different ODs were the following: 4.2×107 cfu/ml (OD550=0.1), 1.7×108 cfu/ml (OD550=0.5), 8.0×108 cfu/ml (OD550=1.0) and 1.3×109 cfu/ml (OD550=2.0).","Preservation yield of E. coli harvested at different growth stages and loaded with 10% MAG is shown in Table 10 and on FIG.","8.TABLE 10 Preservation Yield (Day 0) Stability (Day 7 at RT) Stability (Day 14) Sample % % % Description Cfu/ml Survival Cfu/ml Survival Cfu/ml Survival OD = 0.1, 1.6 × 107 +/− 10.9 +/− 3.3 8.9 × 106 +/− 6.1 +/− 1.0 1.1 × 107 +/− 7.0 +/− 1.9 control 4.8 × 106 1.4 × 106 2.8 × 106 OD = 0.1, 1.6 × 107 +/− 11.3 +/− 0.9 1.2 × 107 +/− 8.2 +/− 0.8 1.5 × 107 +/− 9.9 +/− 2.2 loaded 1.3 × 106 1.2 × 106 3.2 × 106 OD = 0.5, 2.6 × 108 +/− 10.7 +/− 0.6 2.5 × 108 +/− 10.0 +/− 1.3 1.8 × 108 +/− 7.3 +/− 0.3 control 1.5 × 107 3.3 × 107 8.3 × 105 OD = 0.5, 5.5 × 108 +/− 22.2 +/− 2.9 5.3 × 108 +/− 21.6 +/− 4.7 4.3 × 108 +/− 17.4 +/− 1.2 loaded 7.2 × 107 1.2 × 108 2.9 × 107 OD = 1.0, 1.3 × 109 +/− 15.1 +/− 3.7 1.4 × 109 +/− 16.2 +/− 2.8 1.6 × 109 +/− 18.4 +/− 4.6 control 3.3 × 108 2.5 × 108 4.0 × 108 OD = 1.0, 2.3 × 109 +/− 26.3 +/− 4.7 3.2 × 109 +/− 36.1 +/− 7.5 2.9 × 109 +/− 33.0 +/− 5.7 loaded 4.2 × 108 6.5 × 108 4.9 × 108 OD = 2.0, 3.5 × 109 +/− 49 +/− 5.4 4.1 × 109 +/− 57.5 +/− 6.8 4.3 × 109 +/− 60.3 +/− 11.5 control 3.8 × 108 4.8 × 108 8.2 × 108 OD = 2.0, 2.8 × 109 +/− 38.8 +/− 5.8 2.1 × 109 +/− 29.9 +/− 3.6 2.5 × 109 +/− 35.3 +/− 7.5 loaded 4.1 × 108 2.6 × 108 5.3 × 108 No significant difference in preservation survival between loaded and unloaded cells were found when the cells were harvested in early logarithmic stage (at OD005=0.1) before loading with 10% MAG.","In contrast, the cells harvested in mid-log or late-log growth stage and loaded with MAG preserved at higher yield and were significantly more stable than unloaded controls.","The stationary cells, loaded or unloaded, preserved at higher yield than loaded cells.","High preservation survival in stationary cells could be attributed to the presence of intracellular protective compounds.","The importance of the growth stage for efficient loading of MAG is clearly illustrated on FIG.","8.Regardless of a mechanism of MAG accumulation, significant difference in preservation survival and stability between control and loaded cells was obtained only in early- and mid-logarithmic cells suggesting that the loading process was growth-stage dependent.","Based on the data from this example, the following conclusions were reached: 1.Efficiency of loading the cells with MAG is highly dependent upon growth stage of bacteria.","2.In vitro loading with MAG significantly increased preservation yield and stability in mid-log and late-log cells.","3.Loading with MAG resulted in a decrease in preservation survival in stationary cells.","4.Early-log cells preserved at lower yield and were less stable compared to the cells in later growth stage.","5.Stationary cells were the most stable regardless of loading.","EXAMPLE 10 Chemically competent E. coli XL10-Gold cells (1.5 L; Stratagene) in the form of a bacterial pellet were preserved using foam formation.","The cells were concentrated 50-fold by resuspending the pellets in 30 ml of transformation buffer (Stratagene) and processed in the following manner.","This example provides method steps which differ than those previously disclosed.","Therefore, the methods of this example are included herein in their entirety.","Methods (1) Cell density: To determine the cell density in the material, the cells (2×0.1 ml) were diluted in SM buffer (Stratagene) and plated in duplicate (0.1 ml) on L-agar plates.","(2) Transformation of fresh cells: 100 l of cells+0.1 ng or 0.01 ng of pUC18 plasmid DNA (Stratagene) were mixed and incubated on ice for 30 minutes.","When 30 minutes elapsed, the mixtures were transferred in a 42° C. water bath and subjected to a “heat shock” for 30 seconds.","The mixtures were then incubated on ice for 2 minutes and 1 ml of NZY medium (Stratagene) was added.","The mixtures were incubated at 37° C. for 60 minutes with shaking (200 rpm).","The cells were plated on L-agar plates with 100 g/mi ampicilin (50, 100, 200, and 400 l aliquots of undiluted mixtures) for determination of transformation efficiency.","The plates were incubated overnight at 37° C. (3) Freezing of the fresh cells: 10×0.15 ml of the original concentrate were aliquoted in Eppendorf tubes, mixed with 10% DMSO, and frozen at −80° C. Preservation yield and stability in the frozen cells was determined after storage at −80° C. for 3 and 7 days.","(4) Preservation of electrocompetent cells by foam formation: The cells were split in two aliquots and mixed either with preservation solution “A” (22.5% 2:1 sucrose:raffinose) or with solution “B”(22.5% 4:1 sucrose:MAG) 43×0.5 ml of the mixture “A”was aliquoted in 43 sterile 5 ml vials.","Similarly, 9×0.5 ml of the mixture “B” was aliquoted in 9 sterile 5 ml vials.","Vials containing the two preservation mixtures were kept on ice until preservation.","Bacterial survival in the preservation mixtures was determined by plating appropriate dilutions on L-agar plates.","The plates were incubated overnight at 37° C. (5) Drying: Preservation mixtures were dried overnight by foam formation.","Vials containing the mixture “A” were split in two groups and stored at 4° C. and at room temperature (RT).","Vials containing the mixture “B” were stored at RT only.","Stability of the preserved material was initially evaluated after 7 days of storage and will be monitored over time.","(6) Determination of preservation yield: The preserved cells were rehydrated at RT with 0.5 ml of transformation buffer (STB) (Stratagene) or transformation buffer (UTB) (Universal Preservation Technologies).","Stratagene's transformation buffer is available commercially.","UTB buffer comprises: 10 mM MOPS, 75 mM calcium chloride, 10 mM rubidium chloride and 10% sucrose.","Appropriate dilutions were plated on L-agar plates and the plates were incubated at 37° C. overnight.","(7) Transformation of the preserved cells: The preserved cells were transformed in the same manner as the fresh cells.","Results Chemically competent E. coli XL10-Gold cells (Stratagene) were preserved by the foam formation and by freezing at −80° C. The cells were preserved in two different solutions and rehydrated with two different buffers in order to determine possible effects of these parameters on the preservation yield.","Preservation yield and stability of E. coli XL10-Gold cells were determined and presented in Table 11.TABLE 11 Preservation yield (Day 0) Stability (Day 7) Cells Cfu/ml % Cfu/ml % Fresh 1.0 × 1010 +/− 100 N/A N/A 2.6 × 108 Frozen 2.5 × 109 +/− 24.1 +/− 1.4* 2.5 × 109 +/− 1.1 × 108 24.4 +/− 1.0 1.5 × 108* Foam Formation Preserved PS “A”, 9.5 × 107 +/− 0.9 +/− 0.2 4° C. RT 4° C. RT UTB 2.1 × 107 3.9 × 107 +/− 1.0 × 107 3.5 × 107 0.38 +/− 0.34 +/− 0.10 +/− 2.3 × 107 0.22 PS “B”, 3.4 × 106 +/− 0.03 +/− 0.02 4° C. RT 4° C. RT UTB 1.9 × 106 N/A 1.8 × 105 N/A <0.01 +/− 9.3 × 104 PS “A”, 1.1 × 108 +/− 1.1 +/− 0.3 N/D N/D STB 2.8 × 107 PS “B”, 2.0 × 106 +/− 0.02 +/− 0.00 N/D N/D STB 2.5 × 105 PS, preservation solution UTB, transformation buffer (Universal Preservation Technologies) STB, transformation buffer (Stratagene) *Day 1 at −80° C. N/A, not applicable N/D, not determined Preservation yield of cells dried by foam formation were approximately 0.9% (Table 11).","The yield was significantly lower than preservation yield of the frozen cells.","Preservation yield in the cells preserved in solution “A” and rehydrated with UTB was comparable to that in the cells preserved in the same solution and rehydrated with STB.","Similar findings were found for the cells preserved in solution “B”.","However, overall preservation yield was significantly higher (>30-fold) when the cells were preserved in solution “A” compared to that obtained with solution “B” regardless of the rehydration buffer.","After 7 days of storage at 4° C. and at RT, loss in viability of about ½ Log was observed in the material preserved in solution “A”.","Also, a loss of about 1 Log was observed in the material preserved in solution “B”.","There was no significant difference in stability of the material preserved in either solution at the two temperatures.","No loss in viability was observed in frozen cells after 7 days at −80° C. Transformation efficiency of the preserved material is presented in Table 12.The cells were transformed as described in the methods section of this example.","TABLE 12 Transformation Efficiency* Sample Description (Cfu/□g pUC18) Fresh 7.4 × 106 +/− 4.9 × 105 Frozen 3.1 × 106 +/− 3.7 × 105 Foam formation preserved (Day 0) PS “A”, UTB 7.6 × 106 +/− 1.2 × 106 PS “B”, UTB 8.0 × 105 +/− 1.2 × 105 PS “A”, STB 5.9 × 105 +/− 8.8 × 104 PS “B”, STB 3.8 × 104 +/− 1.7 × 104 Frozen (Day 4 at −80° C.) 1.1 × 106** Foam formation preserved (Day 7) PS “A”, UTB, stored at 4° C. 3.5 × 106 +/− 9.2 × 105 PS “A”, UTB, stored at RT 4.0 × 106 +/− 4.6 × 105 PS “B”, UTB, stored at RT 1.0 × 106 +/− 3.6 × 105 PS “A”, STB N/D PS “B”, STB N/D *Transformation efficiencies reported were obtained with 0.1 ng of pUC18.No significant difference was observed when 0.01 ng of pUC18 was used instead of 0.1 ng.","**Only one sample was plated (0.4 ml) and a standard deviation could not be determined.","N/D, not determined.","Transformation efficiency in the cells preserved by foam formation was comparable to that in the fresh and frozen cells (Table 12).","The cells preserved in solution “A” and rehydrated with transformation buffer (Universal Preservation Technologies) had the highest efficiency after drying.","Storage at 4° C. or at RT did not compromise transformation efficiency of the preserved cells.","There was no difference in the efficiency of the preserved cells stored for 7 days compared to the cells immediately after drying.","The feasibility of preservation of chemically competent E. coli XL10-Gold cells was clearly demonstrated by the results discussed above.","Differences in yield were observed for the cells preserved in the two different preservation solutions (“A” and “B”) evaluated in this experiment.","There was no difference in the recovery of the cells preserved in solution “A” or “B” with respect to which rehydration solution was used (UTB or STB buffer).","Overall, preservation survival in the cells dried by the foam formation was lower than for frozen cells (0.9-1.1% versus 24.1%).","Chemically competent cells preserved in this example were harvested in a mid-logarithmic growth stage.","Some initial loss in viability of the preserved cells was observed after 7 days of storage at either RT or 4° C. It is not unusual for cells preserved in logarithmic growth stage without desiccation protectants, like chemically competent cells used in the present example, to lose some stability after initial storage.","Other protective solutions can be used to modulate stability of the preserved material in this example.","Transformation efficiency in the preserved cells was comparable to that in the fresh and frozen cells.","In contrast to preservation survival, where preservation solution was the factor critical to the recovery of the preserved cells, both preservation and rehydration solutions were critical parameters affecting the efficiency of the preserved cells.","In addition, the cells rehydrated with buffer UTB transformed at efficiencies at least 10-fold higher than the cells rehydrated in buffer STB.","Combination of preservation solution “A” and rehydration buffer (Universal Preservation Technologies) resulted in the cells which had the maximal transformation efficiency (7.6×106 cfu/g) after drying.","Transformation efficiency in the preserved cells remained unchanged after storage for 7 days at 4° C. or at RT.","In conclusion, chemically competent E. coli XL10-Gold cells were successfully preserved by foam formation and remained fully competent after preservation and a short-term storage.","EXAMPLE 11 Electrocompetent E coli XL1Blue cells (3 L; Stratagene) in the form of a bacterial pellet were preserved by the foam formation process.","The cells were concentrated 100-fold by resuspending the pellets in 30 ml concentrating solution (Universal Preservation Technologies) and processed in the following manner.","This example provides method steps which differ than those previously disclosed.","Therefore, the methods of this example are included in their entirety.","Methods (1) Cell density: To determine the cell density in the material, the cells (2×0.1 ml) were diluted in SM buffer (Stratagene) and plated in duplicate (0.1 ml) on L-agar plates.","(2) Electroporation of fresh cells: 40 l of cells+0.1 ng of pUC18 plasmid DNA (Stratagene) were mixed, transferred into 0.1 cm BioRad cuvette (Stratagene) and electroporated at E=1.7 kV, R=200 Ohm, C=25 F =4-5 msec).","After the pulse, 1 ml of SOC medium (Stratagene) was added to the cells and the mixture was incubated at 37° C. for 60 minutes with shaking (250 rpm).","The cells were plated on L-agar plates with no antibiotic (for determination of electroporation survival) and on L-agar plates with 100 g/ml ampicilin (for determination of electroporation efficiency).","The plates were incubated overnight at 37° C. (3) Freezing of the fresh cells: 10×0.1 ml of the original concentrate was aliquoted in Eppendorf tubes, mixed with 10% DMSO, and frozen at −80° C. Preservation yield and stability in the frozen cells were determined after storage at −80° C. for 1 and 12 days.","(4) Preservation of electrocompetent cells by foam formation: 3.6 ml of the cells were removed from the 30 ml of the material, centrifuged, and the pellet was resuspended in preservation solution #1 (Universal Preservation Technologies).","The mixture was aliquoted in 16×1.2 ml vials (0.2 ml per vial).","Similarly, 24.8 ml of the remaining cells were centrifuged and the pellet was resuspended in preservation solution #2 (Universal Preservation Technologies).","The mixture was aliquoted in 16×1.2 ml vials (0.2 ml per vial) and in 43×5 ml vials (0.5 ml per vial).","Vials containing the two preservation mixtures were kept on ice until preservation.","Bacterial survival in preservation mixtures was determined by plating appropriate dilutions on L-agar plates.","The plates were incubated overnight at 37° C. (5) Drying: Preservation mixtures were dried overnight by foam formation.","Vials containing the preserved cells were stored at 4° C. and at room temperature (RT).","The 1.2 ml vials were stored at 4° C. The 5 ml vials were divided in two groups and stored at RT and at 4° C. Stability of the preserved material was initially evaluated after 11 days of storage.","(6) Determination of preservation yield: The preserved cells (1.2 ml vials, preservation solutions 1&2; 5 ml vials, preservation solution 2) were rehydrated at room temperature (RT) in solution “A” (Universal Preservation Technologies).","0.5 ml of the solution was added to rehydrate the cells in 5 ml vials, 0.2 ml was used for rehydration of the cells in 1.2 ml vials.","In addition, several 5 ml vials (material preserved in solution 2) were rehydrated with solution “B” (Universal Preservation Technologies).","Appropriate dilutions were plated on L-agar plates and the plates were incubated at 37° C. overnight.","(7) Electroporation of preserved cells: The preserved cells were electroporated in the same manner as the fresh cells.","Results Electrocompetent E. coli XL1Blue cells (Stratagene) were preserved by foam formation and by freezing at −80° C. The cells were preserved in two different solutions, in two vial sizes, and rehydrated with two different solutions in order to determine a possible effect of these parameters on preservation yield.","Preservation yield and stability of the E. coli XL1Blue cells were determined and presented in Table 13.TABLE 13 Stability (Day 12 frozen; Day 11 Foam Preservation yield (Day 0) Formation Preserved) Cells Cfu/ml % Cfu/ml % Fresh 6.5 × 109 +/− 1.8 × 109 100 N/A N/A Frozen N/D N/D 1.2 × 109 +/− 1.1 × 108 18.6 +/− 1.7 Foam Formation Preserved 1.2 ml vial, 2.6 × 108 +/− 1.7 × 107 4.0 +/− 0.3 N/D N/D PS#1, RS “A” 1.2 ml vial, 2.6 × 108 +/− 3.3 × 107 4.0 +/− 0.5 N/D N/D PS#2, RS “A” 5 ml vial, 3.5 × 108 +/− 5.3 × 107 5.3 +/− 0.8 N/D N/D PS#2, RS “A” N/D N/D 5 ml vial, 5.1 × 108 +/− 1.5 × 108 7.9 +/− 2.3 4° C. RT 4° C. RT PS#2, RS “B” 4.4 × 108 +/− 6.6 × 107 3.2 × 108 +/− 6.1 × 107 6.8 +/− 1.0 5.1 +/− 1.0 PS, preservation solution RS, rehydration solution N/A, not applicable N/D, not determined Preservation yield of foam formation dried cells was low, ranging from 4-8% (Table 13).","No significant difference was found in preservation yield when the cells were preserved in two different solutions (PS#1 and PS#2) or in different size vials (1.2 ml or 5 ml) and rehydrated in solution “A” (Universal Preservation Technologies).","Preservation yield was slightly higher when the cells preserved in 5 ml vials were rehydrated in solution “B” (Universal Preservation Technologies).","The preserved cells were stable for 11 days at 4° C. and at RT.","There was no significant difference in the stability of the preserved material at the two temperatures.","Electroporation survival and efficiency of the preserved material are presented in Table 14.The cells were electroporated by the protocol as described previously in the methods section of this example.","TABLE 14 Electroporation Electroporation Survival Efficiency Sample Description (%) (Cfu/□g pUC18) Fresh 19.3 +/2.1 8.7 × 107 +/− 5.6 × 106 Frozen (Day 1 at −80° C.) N/D 1.8 × 106 +/− 3.5 × 105 Frozen (Day 12 at −80° C.) 19.7 +/− 1.6 1.2 × 107 +/− 2.8 × 106 Foam Formation Preserved (Day 0) 1.2 ml vial, PS#1, RS “A” 18.1 +/− 5.9 2.5 × 105 +/− 1.3 × 105 1.2 ml vial, PS#2, RS “A” 24.1 +/− 7.8 9.5 × 105 +/− 5.2 × 105 5 ml vial, PS#2, RS “A” 16.0 +/6.5 2.0 × 105 +/− 1.6 × 105 5 ml vial, PS#2, RS “B” 5.6 +/0.4 7.3 × 105 +/− 3.3 × 105 Foam Formation Preserved (Day 11 at 4° C.) 5 ml vial, PS#2, RS “B” 8.9 +/− 2.0 1.0 × 106 +/− 1.7 × 105 Foam Formation Preserved (Day 11 at RT) 5 ml vial, PS#2, RS “B” 7.8 +/− 1.7 6.8 × 105 +/− 1.7 × 105 N/D, not determined Except for samples preserved in solution #2 (PS#2) in 5 ml vials and rehydrated in solution “B”, there was no significant difference in electroporation survival of the preserved E. coli XL1Blue cells at Day 0 compared to the fresh cells (Table 14).","In addition, there was no significant difference in electroporation survival with respect to a vial size in the cells preserved in solution #1.The cells preserved in solution #2 in 5 ml vials and rehydrated in solution “B” survived electroporation at a lower rate than the cells rehydrated in solution “A”.","Electroporation survival in the stored cells was determined for the sample preserved in solution #2 in 5 ml vials and rehydrated in solution “B”.","There was no difference in electroporation survival after storage at 4° C. or at RT for 11 days compared to that at Day 0 (Table 2).","Electroporation efficiency in the preserved cells was lower compared to that in the fresh or frozen cells (Table 14).","When the cells were preserved in 5 ml vials and rehydrated in solution “B”, electroporation efficiency was higher than in the cells preserved under identical conditions and rehydrated in solution “A”.","Storage at 4° C. or at RT did not compromise electroporation efficiency of the preserved cells.","There was no difference in electroporation efficiency of the preserved cells stored for 11 days compared to the cells immediately after drying.","Based on the data from this example, the following conclusions were made: 1.The feasibility of preservation of electrocompetent E. coli XL1Blue cells was confirmed in the experiment discussed in this example.","2.With respect to formulation of preservation solution, no significant difference in the preservation yield was observed when the cells were preserved in two solutions evaluated in this experiment.","3.The preserved electrocompetent cells were stable at 4° C. and at RT for 12 days.","No significant difference in stability was observed at the two temperatures.","4.Electroporation survival in the preserved cells was comparable to that in the fresh cells and was not compromised by storage at RT or at 4° C. 5.With the preservation and rehydration solutions used in this study, arcing was not a problem.","Also, the formulations used for preservation and rehydration in this example resulted in an overall increase in the transformation efficiency of the preserved cells compared to the efficiency obtained in a similar experiment performed previously (7.3×105 cfu/ g pUC18 DNA compared to 5.0×104 cfu/ g).","Electroporation efficiency in the preserved cells obtained in the experiment presented in this report (7.3×105 cfu/ g) was slightly lower than in the frozen cells (1.8×106 cfu/ g).","In a separate study, we found that the concentration of sugars used in preservation and rehydration solutions has an inhibitory effect on electroporation efficiency when the cells were electroporated at a high voltage (1.7 kV).","Therefore, to achieve the maximal efficiency of the cells, both preservation conditions (solutions to be used, drying protocols, rehydration solutions, etc.)","and the conditions for electroporation (voltage, pulse duration, etc.)","need to be optimized.","Although the invention has been described in detail for the purposes of illustration, it is understood that such detail is solely for that purpose, and variations can be made therein by those skilled in the art without departing from the spirit and scope of the invention which is defined by the following claims.","All references referred to above are hereby incorporated by reference."]],"string":"[\n [\n \"Preservation of competent cells at ambient temperatures\",\n \"The present invention relates to techniques for loading cells with desiccation protectants comprising non-metabolizable and non-reducing carbohydrate analogs.\",\n \"Specifically, competent bacterial cells can be preserved using the loading techniques provided herein.\"\n ],\n [\n \"1.A method of preserving competent bacterial cells for storage at ambient temperatures, comprising: incubating said cells with a nonmetabolizable and non-reducing carbohydrate analog, wherein said incubation results in accumulation of said analog in said cells; and drying said cells by foam formation.\",\n \"2.The method of claim 1, wherein said nonmetabolizable and non-reducing carbohydrate analog is α-methyl-glucoside (MAG) or 2-deoxyglucose (2-DOG).\",\n \"3.The method of claim 1, wherein said cells are gram negative bacteria.\",\n \"4.The method of claim 1, wherein said cells are Escherichia coli (E. coli).\",\n \"5.The method of claim 1, wherein said cells are electrocompetent.\",\n \"6.The method of claim 1, wherein said cells are chemically competent.\",\n \"7.The chemically competent cells of claim 6, wherein competence is achieved by mixing said cells with CaCl2 or RbCl.\",\n \"8.The method of claim 1, further comprising rehydrating the dried cells by contacting said cells with a solution comprising at least one member of the group consisting of carbohydrates, mono-valent cations, divalent cations, organic buffers, and water.\",\n \"9.The method of claim 8, wherein said carbohydrate is sucrose.\",\n \"10.The method of claim 8, wherein said cations are calcium or rubidium.\",\n \"11.The method of claim 1, wherein said carbohydrate analog is administered to said cells at concentrations of 0.1%-50% of total preservation solution.\",\n \"12.The method of claim 1, wherein said carbohydrate analog is administered to said cells at concentrations of 5%-15% of total preservation solution.\",\n \"13.The method of claim 1, wherein said logarithmic cells are mid-logarithmic cells.\",\n \"14.The method of claim 1, wherein said logarithmic cells are late-logarithmic cells.\",\n \"15.The method of claim 1, wherein said logarithmic cells are harvested at OD550 between 0.1-2.0.16.The method of claim 1, wherein said logarithmic cells are harvested at OD550 between 0.3-1.0.17.The method of claim 1, wherein said logarithmic cells are harvested at OD550 at 0.5.18.The method of claim 1, wherein said incubation is conducted at 0° C.-60° C. 19.The method of claim 1, wherein said incubation is conducted at 20° C.-40° C. 20.The method of claim 1, wherein said incubation is conducted at 30° C.-40° C. 21.The method of claim 1, wherein said incubation is conducted for 0-60 minutes.\",\n \"22.The method of claim 1, wherein said incubation is conducted for 2.5-60 minutes.\",\n \"23.The method of claim 1, wherein said incubation is conducted for 30-60 minutes.\",\n \"24.The method of claim 1, wherein said carbohydrate analogs are actively transported into said logarithmic cells.\",\n \"25.The method of claim 1, wherein said logarithmic cells are grown in the presence of glucose prior to preservation in order to prime said cells for active transport.\",\n \"26.The method of claim 25, wherein said logarithmic cells are grown in growth solution comprising 0.001%-50% glucose.\",\n \"27.The method of claim 25, wherein said logarithmic cells are grown in growth solution comprising 0.050%-5% glucose.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>1.Field of the Invention The present invention relates to loading techniques which can be used in conjunction with biological preservation systems.\",\n \"Specifically, competent cells can be preserved using the loading techniques provided herein.\",\n \"2.Description of the Related Art Routine procedures in biotechnological laboratories involve daily use of various competent cells for cloning, propagation and preparation of plasmid DNA, construction of genomic libraries, protein expression, and mutagenesis.\",\n \"Different kinds of competent cells are available commercially, including bacterial, insect, yeast and mammalian cell lines.\",\n \"Various strains of the gram-negative bacterium Escherichia coli ( E coli ) are one of the most extensively used competent cells.\",\n \"Depending on the method of transformation that will be applied, competent cells can be prepared as chemically competent (to be transformed by a heat pulse) or as electrocompetent (to be transformed by an electrical pulse).\",\n \"Chemically competent cells are readily transformable only in the early logarithmic growth stage.\",\n \"However, early logarithmic cells are extremely sensitive to various stresses, including osmotic and thermal stress, making their preservation difficult.\",\n \"As an example, preservation survival of <1% in dried logarithmic E. coli cells was recently reported by D. Billi et al.\",\n \"(Billi, D., D. J. Wright, R. F. Helm, T. Prickett, M. Potts, and J. H. Crowe.\",\n \"2000.Engineering desiccation tolerance in Escherichia coli.\",\n \"Appl.\",\n \"Environ.\",\n \"Microbiol.\",\n \"66:1680-1684).\",\n \"In contrast to chemically competent cells, electrocompetent cells can be transformed in later logarithmic stage and are, therefore, more amenable to preservation.\",\n \"Regardless of the nature of the competent cells and their final use, the cells must be constantly held at subzero temperatures.\",\n \"To allow efficient use, competent cells are often prepared in large quantities and then preserved and stored.\",\n \"Present methodology for the preservation of competent cells includes mixing with different cryoprotectants, for example glycerol, sucrose, dimethly sulfoxide (DMSO), and freezing at −80° C. Because presently available preservation methods include freezing at −80° C., competent cells are routinely subjected to damage by osmotic stress and by exposure to freezing.\",\n \"Commercially available competent cells are sold individually or as components of various kits developed in order to improve efficiency and accuracy of many routine procedures conducted in research laboratories.\",\n \"Most of the kit components, except competent cells, can be stored in laboratory refrigerators or at room temperature.\",\n \"Therefore, the necessity to maintain competent cells at sub-zero temperatures imposes significant burden on distribution of the material and its subsequent storage and use (significant laboratory freezer space is often allocated for the storage of competent cells).\",\n \"Also, the need for sub-zero temperatures prevents use of competent cells and kits containing the cells in various facilities where freezers are not available.\",\n \"Therefore, the demand for a technology that will alleviate the need for handling and storing of competent cells at sub-zero temperatures is certainly present.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The present invention discloses loading techniques which can be used in conjunction with biological preservation systems.\",\n \"Specifically, methods for preserving competent cells, such as E. coli, at ambient temperatures are disclosed.\",\n \"Loading techniques described herein increase desiccation tolerance which allows preservation by foam formation.\",\n \"In one embodiment of the present invention, a method for preserving competent cells for storage at ambient temperatures is disclosed.\",\n \"In the methods described therein, cells are incubated with sugar solution and dried by foam formation.\",\n \"The sugar solution used can be any of a-methyl-glucoside (MAG), 2-deoxyglucose (2-DOG), sucrose, raffinose, or glucose.\",\n \"In a preferred embodiment, cells contemplated for preservation by the present invention include gram negative bacteria and, specifically, E. coli.\",\n \"In another embodiment of the present invention, cells may be electrocompetent or chemically competent.\",\n \"In a preferred embodiment, competence of chemically competent cells is achieved by mixing cells with CaCl 2 or RbCl.\",\n \"In a further embodiment of the present invention, the preserved cells of the present invention may be rehydrated.\",\n \"In a preferred embodiment, the rehydration solution may consist of carbohydrates, mono-valent cations, divalent cations, organic buffers, and water.\",\n \"In an especially preferred embodiment, sucrose is a preferred carbohydrate.\",\n \"Additionally, calcium and rubidium are preferred cations.\",\n \"In another aspect of the invention, methods which enhance desiccation tolerance in logarithmic cells are disclosed.\",\n \"In the methods described therein, cells are incubated with nonmetabolizable and non-reducing carbohydrate analogs.\",\n \"Preferred analogs of the invention include MAG or 2-DOG.\",\n \"In a further embodiment of the invention, the carbohydrate analog is administered at concentrations of 0.1%-50% of the total preservation solution.\",\n \"In an especially preferred embodiment of the invention, the carbohydrate analog is administered at concentrations of 5%-15% of the total preservation solution.\",\n \"The logarithmic cells of the invention are mid or late logarithmic cells.\",\n \"Furthermore, in a preferred embodiment, the logarithmic cells are harvested at OD 550 between 0.1-2.0.In another embodiment, the logarithmic cells are harvested at OD 550 between 0.3-1.0.In an especially preferred embodiment, the logarithmic cells are harvested at OD 550 at 0.5.In a further embodiment of the invention, incubation is conducted at 0° C.-60° C. In another embodiment of the invention, incubation is conducted at 20° C.-40° C. In an especially preferred embodiment of the invention, incubation is conducted at 30° C.-40° C. Furthermore, in another aspect of the invention, incubation is conducted for 0-60 minutes.\",\n \"In another aspect of the invention, incubation is conducted for 2.5-60 minutes.\",\n \"In an especially preferred embodiment, incubation is conducted for 30-60 minutes.\",\n \"In another aspect of the invention, the carbohydrate analogs of the invention are actively transported into logarithmic cells.\",\n \"Additionally, logarithmic cells may be grown in the presence of glucose prior to preservation to prime cells for active transport.\",\n \"The growth solution of the invention may contain 0.001%-50% glucose.\",\n \"In an especially preferred embodiment of the invention, the growth solution may contain 0.05%-5% glucose.\",\n \"According to the present invention, preservation yield of cells dried by foam formation may be increased by incubating cells with sugar solution prior to subjecting the cells to preservation conditions.\"\n ],\n [\n \"BACKGROUND OF THE INVENTION 1.Field of the Invention The present invention relates to loading techniques which can be used in conjunction with biological preservation systems.\",\n \"Specifically, competent cells can be preserved using the loading techniques provided herein.\",\n \"2.Description of the Related Art Routine procedures in biotechnological laboratories involve daily use of various competent cells for cloning, propagation and preparation of plasmid DNA, construction of genomic libraries, protein expression, and mutagenesis.\",\n \"Different kinds of competent cells are available commercially, including bacterial, insect, yeast and mammalian cell lines.\",\n \"Various strains of the gram-negative bacterium Escherichia coli (E coli) are one of the most extensively used competent cells.\",\n \"Depending on the method of transformation that will be applied, competent cells can be prepared as chemically competent (to be transformed by a heat pulse) or as electrocompetent (to be transformed by an electrical pulse).\",\n \"Chemically competent cells are readily transformable only in the early logarithmic growth stage.\",\n \"However, early logarithmic cells are extremely sensitive to various stresses, including osmotic and thermal stress, making their preservation difficult.\",\n \"As an example, preservation survival of <1% in dried logarithmic E. coli cells was recently reported by D. Billi et al.\",\n \"(Billi, D., D. J. Wright, R. F. Helm, T. Prickett, M. Potts, and J. H. Crowe.\",\n \"2000.Engineering desiccation tolerance in Escherichia coli.\",\n \"Appl.\",\n \"Environ.\",\n \"Microbiol.\",\n \"66:1680-1684).\",\n \"In contrast to chemically competent cells, electrocompetent cells can be transformed in later logarithmic stage and are, therefore, more amenable to preservation.\",\n \"Regardless of the nature of the competent cells and their final use, the cells must be constantly held at subzero temperatures.\",\n \"To allow efficient use, competent cells are often prepared in large quantities and then preserved and stored.\",\n \"Present methodology for the preservation of competent cells includes mixing with different cryoprotectants, for example glycerol, sucrose, dimethly sulfoxide (DMSO), and freezing at −80° C. Because presently available preservation methods include freezing at −80° C., competent cells are routinely subjected to damage by osmotic stress and by exposure to freezing.\",\n \"Commercially available competent cells are sold individually or as components of various kits developed in order to improve efficiency and accuracy of many routine procedures conducted in research laboratories.\",\n \"Most of the kit components, except competent cells, can be stored in laboratory refrigerators or at room temperature.\",\n \"Therefore, the necessity to maintain competent cells at sub-zero temperatures imposes significant burden on distribution of the material and its subsequent storage and use (significant laboratory freezer space is often allocated for the storage of competent cells).\",\n \"Also, the need for sub-zero temperatures prevents use of competent cells and kits containing the cells in various facilities where freezers are not available.\",\n \"Therefore, the demand for a technology that will alleviate the need for handling and storing of competent cells at sub-zero temperatures is certainly present.\",\n \"SUMMARY OF THE INVENTION The present invention discloses loading techniques which can be used in conjunction with biological preservation systems.\",\n \"Specifically, methods for preserving competent cells, such as E. coli, at ambient temperatures are disclosed.\",\n \"Loading techniques described herein increase desiccation tolerance which allows preservation by foam formation.\",\n \"In one embodiment of the present invention, a method for preserving competent cells for storage at ambient temperatures is disclosed.\",\n \"In the methods described therein, cells are incubated with sugar solution and dried by foam formation.\",\n \"The sugar solution used can be any of a-methyl-glucoside (MAG), 2-deoxyglucose (2-DOG), sucrose, raffinose, or glucose.\",\n \"In a preferred embodiment, cells contemplated for preservation by the present invention include gram negative bacteria and, specifically, E. coli.\",\n \"In another embodiment of the present invention, cells may be electrocompetent or chemically competent.\",\n \"In a preferred embodiment, competence of chemically competent cells is achieved by mixing cells with CaCl2 or RbCl.\",\n \"In a further embodiment of the present invention, the preserved cells of the present invention may be rehydrated.\",\n \"In a preferred embodiment, the rehydration solution may consist of carbohydrates, mono-valent cations, divalent cations, organic buffers, and water.\",\n \"In an especially preferred embodiment, sucrose is a preferred carbohydrate.\",\n \"Additionally, calcium and rubidium are preferred cations.\",\n \"In another aspect of the invention, methods which enhance desiccation tolerance in logarithmic cells are disclosed.\",\n \"In the methods described therein, cells are incubated with nonmetabolizable and non-reducing carbohydrate analogs.\",\n \"Preferred analogs of the invention include MAG or 2-DOG.\",\n \"In a further embodiment of the invention, the carbohydrate analog is administered at concentrations of 0.1%-50% of the total preservation solution.\",\n \"In an especially preferred embodiment of the invention, the carbohydrate analog is administered at concentrations of 5%-15% of the total preservation solution.\",\n \"The logarithmic cells of the invention are mid or late logarithmic cells.\",\n \"Furthermore, in a preferred embodiment, the logarithmic cells are harvested at OD550 between 0.1-2.0.In another embodiment, the logarithmic cells are harvested at OD550 between 0.3-1.0.In an especially preferred embodiment, the logarithmic cells are harvested at OD550 at 0.5.In a further embodiment of the invention, incubation is conducted at 0° C.-60° C. In another embodiment of the invention, incubation is conducted at 20° C.-40° C. In an especially preferred embodiment of the invention, incubation is conducted at 30° C.-40° C. Furthermore, in another aspect of the invention, incubation is conducted for 0-60 minutes.\",\n \"In another aspect of the invention, incubation is conducted for 2.5-60 minutes.\",\n \"In an especially preferred embodiment, incubation is conducted for 30-60 minutes.\",\n \"In another aspect of the invention, the carbohydrate analogs of the invention are actively transported into logarithmic cells.\",\n \"Additionally, logarithmic cells may be grown in the presence of glucose prior to preservation to prime cells for active transport.\",\n \"The growth solution of the invention may contain 0.001%-50% glucose.\",\n \"In an especially preferred embodiment of the invention, the growth solution may contain 0.05%-5% glucose.\",\n \"According to the present invention, preservation yield of cells dried by foam formation may be increased by incubating cells with sugar solution prior to subjecting the cells to preservation conditions.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is a bar chart showing preservation survival of electrocompetent E. coli cells preserved by foam formation.\",\n \"FIG.\",\n \"2 is a bar chart showing electroporation survival of electrocompetent E. coli cells preserved by foam formation.\",\n \"FIG.\",\n \"3 is a bar chart showing electroporation efficiency of electrocompetent E. coli cells preserved by foam formation.\",\n \"FIG.\",\n \"4 is a line graph showing the effect of increasing concentration of glucose analogs during in vitro loading on preservation survival of logarithmic E. coli.\",\n \"FIG.\",\n \"5 is a line graph showing the effect of loading temperature on preservation survival of E. coli.\",\n \"FIG.\",\n \"6 is a line graph showing the effect of loading temperature on stability of the preserved E. coli.\",\n \"FIG.\",\n \"7 is a line graph showing the effect of loading time on preservation survival and stability of E. coli cells loaded with 10% MAG at 37° C. FIG.\",\n \"8 is a line graph showing the effect of growth stage on the preservation survival of E. coli loaded with 10% MAG.\",\n \"DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT The present invention discloses loading techniques which can be used in conjunction with biological preservation systems.\",\n \"Specifically, the invention relates to methods of using loading techniques to increase desiccation tolerance that will allow preservation by foam formation.\",\n \"In a preferred embodiment, the methods described herein may be used to preserve E. coli cells.\",\n \"In order to preserve competent cells under conditions less damaging then freezing and to allow their subsequent handling at temperatures above sub-zero, we used the foam formation process.\",\n \"Foam formation technology represents an advanced method for preservation by drying, where the sample temperature is always kept above freezing (U.S. Pat.\",\n \"No.\",\n \"5,766,520).\",\n \"Therefore, the possibility for cell damage by formation of ice-crystals, which is the most common damage during preservation by freezing, is eliminated.\",\n \"However, because drying itself imposes significant osmotic stress on logarithmic cells, cellular desiccation tolerance should be optimized in order to take the full advantage of the foam formation preservation process.\",\n \"We demonstrated that incubation of the cells with an appropriate sugar solution prior to preservation (“loading”), can significantly increase preservation yield of cells dried by foam formation.\",\n \"Herein we describe a method to preserve E coli cells at ambient temperature by the foam formation process and demonstrate that the preserved cells are transformable at efficiency comparable to that of the fresh cells.\",\n \"Preservation of Electrocompetent Cells Preservation of electrocompetent cells in the various working examples included infra were conducted as follows, unless otherwise stated: (1) Preparation of fresh electrocompetent cells: Escherichia coli ATCC strain MM294 cells were grown under standard conditions in L-broth (growth medium for E. coli), at 37° C., with moderate aeration (shaking at 250 rpm).\",\n \"An isolated colony from a fresh streak was inoculated into 5 ml of L-broth and incubated overnight (preculture).\",\n \"The following morning, the preculture was subcultured (1% inoculum) in 100 ml of L-broth+glucose (0.05%-0.5%).\",\n \"The cells were incubated until OD550=0.5 (subculture).\",\n \"At this point, the cells were transferred into 250 ml of L-broth+glucose (0.05%-0.5%) and incubation continued until OD550=0.8.Two flasks with 250 ml of cells were prepared.\",\n \"When the culture OD550 reached 0.8 (cell density approx.\",\n \"5×108 cfu/ml), the cells were harvested by centrifugation at 2,500 rpm (at room temperature) for 10 minutes.\",\n \"The cells (2×200 ml samples) were washed once with an equal volume of PBS buffer or twice with an equal volume of 20% ice-cold sucrose and centrifuged again as described above.\",\n \"The cells were concentrated 100-fold in PBS buffer or in 20% sucrose to cell density of approx.\",\n \"2×1010 cfu/ml.\",\n \"Bacterial counts were determined by diluting cells in PBS buffer and spread plating appropriate dilutions in duplicate on L-agar.\",\n \"The plates were incubated at 37° C. overnight and colonies were counted the following day.\",\n \"(2) Electroporation of fresh E. coli electrocompetent cells: Two aliquots of the concentrated cells (50 μl each) were transferred into pre-chilled Eppendorf tubes (1.5 ml) and 2 μl of pUC18 plasmid DNA (approx.\",\n \"300 ng/μl) were added to the tubes.\",\n \"The [cells+DNA] mixtures were incubated on ice for 5 minutes and transferred into pre-chilled cuvettes for electroporation.\",\n \"The cells were electroporated by using a BTX T-100 electroporator, at 625 or at 800 volts (V) in BTX cuvettes (0.1 cm gap, P/N 610 Disposable Cuvettes) with a time constant “r” equal 5 msec.\",\n \"Under the pulse conditions described above, the cells were subjected to an electrical field “E” equal to 6.25 kV/cm or 8.0 kV/cm.\",\n \"Immediately after the pulse, the cells were transferred into small glass tubes (10×100 mm) with 0.5 ml of “outgrowth medium” (L-broth+20% sucrose+10 mM CaCl2).\",\n \"The electroporated bacteria were incubated at 37° C. with moderate shaking (150 rpm) for 60 minutes.\",\n \"When the 60 minutes period elapsed, the cells were diluted in “outgrowth medium” and duplicate samples were spread plated on L-agar plates with no antibiotic (to determine electroporation survival) and on L-agar plates with 50 μg/ml or 100 μg/ml of ampicillin (selective marker on pUC18 plasmid; to determine electroporation efficiency).\",\n \"The plates were incubated overnight at 37° C. and the colonies were counted the next day.\",\n \"(3) Loading of fresh electrocompetent E. coli cells before preservation: When applicable, one aliquot of the cell concentrate (3.2 ml of cells in PBS buffer, approx.\",\n \"2×1010 cfu/ml) was mixed with 0.8 ml of 50% methyl-glucoside (MAG; 10% final concentration) and incubated at 37° C. for 30 minutes (in vitro loading).\",\n \"A second aliquot (3.2 ml of the cell concentrate) was mixed with 0.8 ml of PBS buffer and incubated on ice for 30 minutes (control, unloaded cells).\",\n \"Similar to loading with MAG, the cells were loaded with 2-deoxyglucose (2-DOG) as described in Example 5 infra.\",\n \"(4) Preservation of electrocompetent E. coli cells: The cells (resuspended in PBS buffer or on 20% sucrose) were mixed with an equal mass of preservation solution consisting of 45% 2:1 sucrose:raffinose or 45% 4:1 sucrose:MAG on ice.\",\n \"Altematively, the cell pellets were resuspended directly in 22.5% sucrose:raffinose or 22.5% 4:1 sucrose:MAG.\",\n \"The 0.5 g aliquots were distributed in 5 ml glass serum vials or 0.2 g aliquots were distributed in 1.2 ml glass serum vials.\",\n \"To determine the survival in preservation solution before drying, cell-containing mixtures were diluted in PBS buffer containing 20% sucrose and appropriate dilutions were spread plated in duplicate on L-agar plates.\",\n \"The samples were dried by foam formation.\",\n \"The mixtures were foamed at 0° C. and kept at that temperature until foam formation occurred.\",\n \"After foam was well formed, shelf temperature was increased to 20° C. and drying continued overnight (at least 12 hours).\",\n \"When applicable, electrocompetent cells were frozen in the following manner.\",\n \"The cell suspension (100 μl) was mixed with 10% DMSO (10 μl) and slowly frozen at −80° C. (5) Assaying the preserved E. coli cells: Dried samples were rehydrated at room temperature.\",\n \"1 ml of rehydration solution (20% sucrose) was added to the 5 ml vials and 0.2 ml was added to the 1.2 ml vials.\",\n \"For the determination of preservation survival, rehydrated cells were diluted and spread plated in duplicate on L-agar plates.\",\n \"Plates were incubated at 37° C. overnight and colonies were counted the following day.\",\n \"(6) Electroporation of dried (preserved) E. coli cells: One or two 50 μl aliquots were removed from each rehydrated vial and transferred to pre-chilled Eppendorf tubes.\",\n \"The cells were mixed with pUC18 plasmid DNA (300 ng/μl) and electroporated using the conditions described previously for the electroporation of fresh electrocompetent cells.\",\n \"Electroporation survival was calculated as the number of viable cells recovered after electrical pulse per total number of electroportated cells.\",\n \"Electroporation efficiency was calculated as the number of transformants (cells which received pUC18 plasmid DNA) per number of cells recovered after electroporation (electroporation survival).\",\n \"When applicable, the electroporation efficiency was also expressed as number of transformants per 1 μg of pUC18 DNA (Cfu/μg).\",\n \"Preservation of Chemically Competent Cells Preservation of chemically competent cells in the various working examples included infra was conducted as follows, unless otherwise stated: (1) Preparation of chemically competent cells: The cells were grown as described for preparation of electrocompetent cells, except that the culture was harvested when OD550 reached 0.5.The cells were chilled on ice and centrifuged as described for electrocompetent cells.\",\n \"The supernatant was removed and the cells were and re-suspend in 30 ml of ice-cold Buffer A [30 mM KOAc, 50 mM MnCl2, 100 mM RbCl, 10 mM CaCl2, 10% sucrose), pH=5.8.The cells were incubated on ice for 120 minutes.\",\n \"When the 120 minutes period elapsed, the cells were centrifuged for 5 minutes at 2,500 rpm in a pre-chilled centrifuge.\",\n \"The supernatant was removed and the cells resuspended in 4 ml of ice-cold Buffer B [10 mM Na MOPS (pH=7.0), 75 mM CaCl2, 10 mM RbCl, 10% sucrose].\",\n \"The cells were kept on ice at all times.\",\n \"(2) Preservation of chemically competent E. coli cells: The cells were transferred into ice-cold 15 ml Falcon tubes.\",\n \"3.5 ml of cells were mixed with 3.5 g of ice-cold preservation solution consisting of 40% 3:1 sucrose:MAG in PBS buffer.\",\n \"The 2×0.1 ml of preservation mixture were removed, serially diluted with PBS buffer and plated in duplicate on agar plates with no antibiotic (“preservation mixture controls”).\",\n \"The 0.5 g aliquots of preservation mixture were distributed into pre-chilled 5 ml sterile glass vials.\",\n \"The mixtures were preserved by foam formation as described for drying of electrocompetent cells.\",\n \"(3) Assaying of the preserved chemically competent E. coli cells: Two vials of the dried cells were transferred on ice and re-hydrated with 0.5 ml of Buffer B.\",\n \"For the determination of preservation survival, rehydrated cells were diluted in buffer B and spread plated in duplicate on L-agar plates.\",\n \"Plates were incubated at 37° C. overnight and colonies were counted the following day.\",\n \"The vials were stored in a refrigerator for future assays.\",\n \"(4) Transformation of chemically competent E. coli cells by “heat shock”: The fresh and the preserved cells were transformed under identical conditions, except that in order to keep the same cell density during the transformation, 0.2 ml of the preserved cells and 0.1 ml of the fresh cells were used.\",\n \"The cells were transformed in the following manner.\",\n \"0.1 ml of the fresh cells or 0.2 ml of re-hydrated content from each vial were transferred into pre-chilled Eppendorf tubes.\",\n \"The pUC18 plasmid DNA (5 μl, approx.\",\n \"500 ng) was added into each Eppendorf tube with cells.\",\n \"The cells and DNA were mixed gently and the mixtures were incubated on ice for 120 minutes.\",\n \"When the 120 minutes period elapsed, the cells were transferred for 90 seconds at 42° C. (“heat shock”).\",\n \"The cells were returned on ice for 2 minutes and then transferred into 5 ml glass tubes containing 5 volumes of L-broth pre-warmed at room temperature.\",\n \"The cells were shaken gently at 37° C. for 60 minutes (150 rpm).\",\n \"Following the 60 minutes incubation at 37° C., 0.1 ml aliquots of the cells were removed from the tubes, diluted with L-broth medium and spread-plated in duplicate on L-agar plates with 50 μg/ml of ampiclin (Ap50).\",\n \"The plates were incubated aerobically at 37° C. overnight.\",\n \"The following working examples illustrate preservation of electrocompetent and chemically competent cells by the use of loading in accordance with the methods of the present invention: EXAMPLE 1 Electrocompetent E. coli MM294 cells were prepared as described previously, except for as follows: In this experiment, the cells were washed twice in ice-cold 20% sucrose and concentrated 100-fold after the second wash. Additionally, four samples were prepared (A, B, C, and D).\",\n \"The cell concentrates were diluted 1:1 with the preservation solution consisting of 45% 2:1 sucrose:raffinose (sample A) or resuspended directly in 22.5% 2:1 sucrose:raffinose (samples B and C).\",\n \"Samples A and B were preserved in 0.5 ml vials (0.5 g fill per vial).\",\n \"Sample C was preserved in 1.2 ml vials (0.2 g fill per vial).\",\n \"Additional solution consisting of 22.5% 4:1 sucrose:MAG was prepared and evaluated for preservation of the cells (sample D).\",\n \"The cell concentrate (“D”) was resuspended directly in 22.5% 4:1 sucrose:MAG and preserved in 1.2 ml vials (0.2 g fill per vial).\",\n \"Dried cells of all samples were rehydrated in 20% sucrose and assayed for stability (1 ml was added to the cells in 5 ml vials, 0.2 ml to the cells in 1.2 ml vials).\",\n \"The preserved cells of sample B were rehydrated with nano-pure water (0% sucrose), 10%, and 20% sucrose and electroporated.\",\n \"Electroporation survival and efficiencies in samples rehydrated with solutions containing different amounts of sucrose were determined.\",\n \"Finally, fresh and dried cells were electroporated at 800V.\",\n \"Electroporation survival and efficiencies were determined and compared.\",\n \"Preservation yield, electroporation survival and efficiencies obtained with the cells in four different samples are shown in Table 1.TABLE 1 Sample Preservation Electroporation Electroporation Description Survival (%) Survival (%) Efficiency (10−3%) Fresh Cells ND 95.8 +/− 17.2 0.50 +/− 0.06 Foam Formation Preserved (Day 0) A 36.8 +/− 2.7 36.6 +/− 3.9 8.1 +/− 0.8 B 21.5 +/− 5.6 70.2 +/− 12.4 2.2 +/− 0.5 C 18.5 +/− 3.2 43.0 +/− 8.1 2.7 +/− 0.2 D 18.7 +/− 2.9 63.9 +/− 11.4 3.7 +/− 0.2 Foam Formation Preserved (Day 11 at 4° C.) A 39.8 +/− 4.5 15.1 +/− 1.7 19.8 +/− 2.5 B 30.2 +/− 3.2 19.9 +/− 7.7 3.3 +/− 1.7 C 22.7 +/− 5.8 19.1 +/− 2.4 3.7 +/− 0.9 D 32.2 +/− 7.8 13.8 +/− 1.0 10.4 +/− 0.9 Foam Formation Preserved (Day 11 at RT°) A 36.2 +/− 6.4 19.5 +/− 1.5 21.1 +/− 5.1 B 25.9 +/− 1.5 25.5 +/− 5.9 4.7 +/− 1.1 C 23.6 +/− 8.6 10.1 +/− 5.6 8.2 +/− 1.6 D 27.6 +/− 6.5 15.0 +/− 6.6 8.2 +/− 1.9 Foam Formation Preserved (Day 20 at 4° C.) A 38.9 +/− 10.9 30.6 +/− 2.0 17.2 +/− 5.5 B N/D N/D N/D C N/D N/D N/D D N/D N/D N/D Foam Formation Preserved (Day 20 at RT°) A 30.3 +/− 2.3 33.9 +/− 2.9 22.8 +/− 8.2 B 17.8 +/− 5.3 22.3 +/− 3.5 5.6 +/− 3.1 C 27.4 +/− 10.4 18.2 +/− 3.6 5.9 +/− 0.8 D 21.7 +/− 0.5 29.9 +/− 6.7 5.1 +/− 0.8 The cell concentrate diluted 1:1 in the preservation solution (A) preserved at a higher yield compared to the concentrate (B) resuspended directly in preservation solution (36.8% versus 21.5%).\",\n \"There was no significant difference in the preservation yield or stability of the preserved samples with respect to the preservation solution used (18.5% in sample C and 18.7% in sample D at Day 0).\",\n \"Regardless of the concentration of the cells in a sample or a vial size, the preserved cells were stable for 20 days at RT and at 4° C. Comparison of the electroporation parameters in preserved and fresh cells revealed that electroporation survival in the preserved cells was lower than in the fresh cells.\",\n \"In contrast, electroporation efficiency in the preserved cells was higher.\",\n \"The preserved cells routinely electroporated at the efficiency 3 to 10 fold higher than the fresh cells.\",\n \"With respect to electroporation parameters in the preserved cells, electroporation survival at Day 0 was higher when the cells were more concentrated (sample B versus sample A).\",\n \"When the cells were preserved in small aliquots (0.2 g in 1.2 ml vial), electroporation survival was higher in the cells preserved in sucrose:MAG (D) compared to the cells preserved in sucrose:raffinose (C).\",\n \"Differences in electroporation survival between different samples were smaller at Day 20 than at Day 0.At Day 0, electroporation efficiency was somewhat higher in the preserved cells in sample A than in sample B.\",\n \"Similar to electroporation survival, the electroporation efficiency was slightly higher at Day 0 in the cells preserved in sucrose:MAG compared to the cells preserved in sucrose:raffinose.\",\n \"At all times (Day 0-Day 20), transformation efficiency was significantly higher in the preserved cells in sample A compared to the cells in other samples.\",\n \"The effect of the sucrose concentration in the rehydration medium on preservation survival and electroporation parameters was evaluated.\",\n \"Relevant data are presented in Table 2.TABLE 2 Preserved cells of sample B rehydrated with different amounts of sucrose Preservation Electroporation Electroporation Sample Description Survival (%) Survival (%) Efficiency (10−3%) 0% sucrose 14.8 +/− 2.9 37.3 +/− 6.3 45.6 +/− 1.5 10% sucrose 32.4 +/− 8.0 35.4 +/− 4.0 10.7 +/− 1.4 20% sucrose 23.4 +/− 2.8 74.5 +/− 8.2 0.9 +/− 0.3 According to data presented in Table 2, the cells rehydrated without sucrose had lower preservation yield than the samples rehydrated with 10% or 20% sucrose (14.8% versus 23-32%).\",\n \"Similar to the preservation survival, the electroporation survival was higher in the preserved cells rehydrated in the solution containing the higher concentration of sucrose (74.5% in 20% sucrose versus 37.3% with no sucrose).\",\n \"Increase in the amount of sucrose in the rehydration solution had an inhibitory effect on electroporation efficiency.\",\n \"The efficiency in the cells rehydrated without sucrose was 45.6% compared to 0.9% in the cells rehydrated with 20% sucrose (Table 2).\",\n \"Based on the data from this example, the following conclusions were reached: 1.Electrocompetent E. coli cells could be successfully preserved by foam formation.\",\n \"2.There was no difference in preservation survival when the cells were preserved in small aliquots (0.2 g in 1.2 ml vial compared to 0.5 g in 0.5 ml vial).\",\n \"3.Preservation survival and stability were slightly better in diluted cells (A versus B).\",\n \"4.There was no significant difference in preservation survival when the cells were preserved in sucrose:raffinose or in sucrose:MAG.\",\n \"5.Electroporation survival in the preserved cells was lower than in the fresh cells.\",\n \"6.Electroporation efficiency in the preserved cells was routinely higher than in the fresh cells.\",\n \"Efficiency was the highest in the cells which were diluted 1:1 with preservation solution.\",\n \"7.Increase in the amount of sucrose in rehydration medium enhanced preservation and electroporation survival, but was inhibitory with respect to the electroporation efficiency.\",\n \"EXAMPLE 2 To enhance bacterial desiccation tolerance, the cells were loaded with 10% MAG.\",\n \"The yield of the preserved electrocompetent E. coli cells, electroporation survival and electroporation efficiency of fresh and preserved cells (loaded and unloaded control) were evaluated and compared (Table 3, FIG.\",\n \"1, FIG.\",\n \"2, and FIG.\",\n \"3).\",\n \"TABLE 3 Electroporation Electroporation Preservation Yield Survival Efficiency Cells Cfu/ml % Cfu/ml % Cfu/ml 10−3% Fresh 2.0 × 1010 N/A 8.6 × 109 43.5 2.1 × 105 2.4 Preserved (Dried) Control 3.8 × 109 19.3 6.3 × 108 16.5 1.6 × 105 25.4 (Treatment A) Loaded 1.1 × 1010 53.5 3.0 × 109 28.6 1.7 × 105 5.6 (Treatment B) N/A, not applicable.\",\n \"The data represents the average of three independent experiments.\",\n \"The preservation yield of the loaded dried electrocompetent cells was significantly higher than the yield in unloaded controls (53.5% versus 19.3%, Table 3).\",\n \"Therefore, loading with MAG ameliorated desiccation damage during foam formation resulting in increased preservation survival.\",\n \"Electroporation survival of dried cells was lower than in fresh cells (Table 3).\",\n \"In dried loaded cells, electroporation survival was higher than in dried control (28.6% versus 16.5%, Table 3).\",\n \"Electroporation efficiency of the preserved loaded cells was somewhat higher than in the fresh cells (5.6×10−3% versus 2.4×10−3%, Table 3).\",\n \"The following conclusions were made based on the data from this example: 1.Loading of the cells with 10% MAG significantly increased preservation survival.\",\n \"2.Electroporation survival in the preserved cells was lower than in the fresh cells.\",\n \"3.Similar to preservation survival, loading of the cells with 10% MAG significantly increased electroporation survival.\",\n \"4.Electroporation efficiency (cfu/ml) in the preserved cells was comparable or slightly higher compared to the fresh cells.\",\n \"EXAMPLE 3 In this example, commercial electrocompetent E. coli cells, in the form of a bacterial pellet, were preserved by the foam formation process.\",\n \"The cells were concentrated 100-fold and preserved as described previously.\",\n \"To increase desiccation tolerance and enhance preservation survival.\",\n \"One aliquot of the cells were loaded in vitro with 10% MAG.\",\n \"The preserved cells were stored at 4° C. In addition to preservation by foam formation, the cells were preserved by slow freezing (with 10% DMSO) at −80° C. Preservation yields and stability in commercial electrocompetent E. coli cells preserved by foam formation and by freezing are presented in Table 4.TABLE 4 Preservation yield** Stability (12 days) Cells Cfu/ml % Cfu/ml % Fresh 1.4 × 1010 +/− 2.8 × 109 100 +/− 19.9 N/A N/A Frozen* 1.1 × 1010 +/− 2.1 × 109 75.6 +/− 14.8 1.2 × 1010 +/− 5.4 × 108 82.7 +/− 3.9 Preserved (Control)* 6.6 × 109 +/− 4.5 × 108 47.7 +/− 3.2 4.9 × 109 +/− 1.5 × 109 35.8 +/− 10.9 Preserved (Loaded) 8.8 × 109 +/− 3.9 × 108 63.6 +/− 2.8 9.5 × 109 +/− 1.1 × 108 68.5 +/− 0.8 *The control cells were also assayed for stability after 67 days at 4° C. Some loss in viability was observed (20 +/− 1% survival, 3 × 109 +/− 2.0 × 108 cfu/ml).\",\n \"**Frozen cells were incubated 72 hours at −80° C.; Foam formation-preserved cells were dried overnight.\",\n \"N/A, not applicable.\",\n \"The control (unloaded) commercial cells dried by foam formation were preserved at a 47.7% yield.\",\n \"The cells were relatively stable at 4° C. (35.8% viability after 12 days and 20% viability after 67 days storage).\",\n \"Commercial cells loaded in vitro with 10% MAG and preserved by foam formation had a yield of 63.6%.\",\n \"These cells were completely stable at 4° C. (68.5% yield after 12 days).\",\n \"The frozen cells were also stable after 12 days of storage at −80° C. Preserved commercial cells (vitrified and frozen) were electroporated as described previously.\",\n \"Electroporation survival and electroporation efficiency in the preserved cells were determined and compared to the same parameters in the fresh cells (Table 5).\",\n \"TABLE 5 *Preserved *Preserved Cells Fresh (Control) (Loaded) Electroporation Survival (Cfu/ml) 6.40 × 109 +/− 1.3 × 109 7.9 × 108 +/− 1.2 × 108 3.2 × 109 +/− 4.2 × 108 (%) 46.2 +/− 9.4 12.2 +/− 1.7 35.7 +/− 4.7 Electroporation Efficiency (Cfu/ml) 1.5 × 105 +/− 4.0 × 104 7.0 × 104 +/− 5.1 × 104 1.9 × 105 +/− 1.7 × 105 (10−3%) 2.4 +/− 0.6 8.8 +/− 6.4 6.1 +/− 5.5 (Cfu/□g DNA) 4.6 × 104 +/− 1.2 × 104 4.3 × 103 +/− 3.1 × 103 1.6 × 104 +/− 1.8 × 102 *Cell density in the preserved cells was {fraction (1/10)} of that in fresh concentrates.\",\n \"These cells were transformed immediately after drying and they were not concentrated before electroporation.\",\n \"Electroporation survival in the preserved cells was lower than in the fresh cells (12.2% or 35.7% versus 46.2%).\",\n \"In contrast, electroporation efficiency in the preserved cells was higher than in the fresh cells (6.1% or 8.8% versus 2.4%).\",\n \"Electroporation survival in preserved control cells was lower than in preserved loaded cells.\",\n \"Electroporation efficiencies were comparable in unloaded and loaded preserved cells (6.1% versus 8.8%).\",\n \"Based on the data from this example, the following conclusions were made: 1.Commercial electrocompetent E. coli cells were successfully preserved by the foam formation process.\",\n \"2.Loading with MAG increased stability of the preserved cells at 4° C. 3.Electroporation survival was higher in fresh cells than in the preserved cells.\",\n \"4.In preserved cells, electroporation survival was higher in loaded cells compared to unloaded control.\",\n \"5.Electroporation efficiency was higher in the preserved cells compared to that in the fresh cells.\",\n \"6.There was no significant difference in electroporation efficiency in MAG-loaded preserved cells versus unloaded preserved controls (MAG was slightly inhibitory).\",\n \"EXAMPLE 4 Chemically competent E. coli MM294 cells were prepared and preserved by foam formation as described previously.\",\n \"Preservation yield and stability of the preserved cells are described in Table 6.TABLE 6 Preservation Yield (Day 0) Stability (290 Days at 4° C.) Cells Cfu/ml % Cfu/ml % Fresh 1.6 × 1010 +/− 6.8 × 109 N/A N/A N/A Preserved 5.1 × 107 +/− 4.0 × 107 0.3 +/− 0.3 1.1 × 107 +/− 4.5 × 106 0.07 +/− 0.03 Chemically competent E. coli MM294 cells were preserved at low yield (0.3%).\",\n \"The cells were harvested in early logarithmic stage and were not loaded with MAG or any other sugar derivative to enhance bacterial desiccation tolerance and the subsequent preservation survival.\",\n \"The preserved cells were relatively stable at 4° C. After 290 days of storage, the observed loss in viability was less than 1 Log (Table 6).\",\n \"Transformation efficiencies of the fresh and the preserved chemically competent cells are presented in Table 7.TABLE 7 Transformation Efficiency Day 0 290 Days at 4° C. Cells Cfu/ml 10−3% Cfu/ml 10−3% Fresh 9.7 × 105 +/− 1.9 × 105 6.1 +/− 1.1 N/A N/A Preserved 6.7 × 102 +/− 2.4 × 102 1.3 +/− 0.5 1.6 × 102 +/− 5.0 × 101 1.5 +/0.5 Transformation efficiency in the preserved cells was somewhat lower compared to the efficiency of the fresh cells (Table 7).\",\n \"Preservation of the cells by foam formation did not compromise bacterial competence in any significant fashion.\",\n \"Transformation efficiency of the preserved cells stored for 290 days at 4° C. was comparable to the efficiency immediately after drying.\",\n \"Based on the data of this example, the following conclusions were reached: 1.Chemically competent E. coli cells could be successfully preserved by foam formation.\",\n \"2.Preservation yield was low because the cells were harvested in early logarithmic phase when they likely had no internal desiccation protectants.\",\n \"3.The cells remained relatively stable upon prolonged storage at 4° C. 4.The preserved cells remained competent after preservation.\",\n \"5.The competence function was not compromised during storage at 4° C. EXAMPLE 5 Accumulation of MAG in vitro enhances preservation survival of logarithmic E. coli cells.\",\n \"Preparation of electrocompetent cells routinely requires harvesting bacterial cultures in late logarithmic growth stage.\",\n \"In contrast, the cells must be harvested in an early or medium logarithmic growth stage for preparation of chemically competent cells.\",\n \"Late logarithmic cells are more tolerant to desiccation than early logarithmic cells.\",\n \"We found that desiccation tolerance of logarithmic E. coli could be significantly enhanced by loading the cells with MAG.\",\n \"We also found that efficiency of loading could be influenced by the following parameters: 1.Choice of carbohydrate 2.Loading temperature 3.Loading time 4.Growth stage of the cells Accumulation of non-metabolizable glucose analogs enhances preservation of logarithmic E. coli cells.\",\n \"Bacterial culture was grown until OD550=0.5 was reached (mid-log growth stage).\",\n \"The cell concentrates were prepared as described previously.\",\n \"Control cells, which were not loaded, were incubated on ice for 30 minutes.\",\n \"Cell concentrates were incubated with different amounts (0.15-1%, 5%, 10% and 15%) of glucose and glucose nonmetabolizable analogs at 37° C. for 30 minutes (loading).\",\n \"These mixtures were preserved as described previously.\",\n \"Mid-logarithmic cells loaded with 0.1%/-1% and 5% of MAG survived drying at 9%, 12% and at 24%, respectively.\",\n \"When cells were loaded with 0.1-1%, 5%, 10% and 15% of 2-DOG, bacteria survived drying at 9-12%, 20%, 16%, and 20%, respectively (FIG.\",\n \"4).\",\n \"Loading with more than 5% of either MAG or 2-DOG in vitro did not improve drying survival compared to the survival obtained when 5% of either compound was used.\",\n \"Control cells, which were not loaded, survived drying at <1%.\",\n \"In contrast to its nonmetabolizable analogs MAG or 2-DOG, glucose had no protective effect on preservation survival of logarithmic cells.\",\n \"Mid-logarithmic cells loaded in vitro with up to 15% of glucose survived drying at 1-3% (FIG.\",\n \"4).\",\n \"There was no significant difference in survival when different amounts of glucose were used for loading.\",\n \"Because glucose was used at concentrations significantly higher than physiological, likely only a portion of the sugar was metabolized.\",\n \"Remaining glucose had no immediate damaging effect on preservation survival.\",\n \"However, because of its high reducing activity, glucose could be detrimental to cells due to a non-enzymatic browning, including the Mallard's reaction.\",\n \"Based on the data of this example, the following conclusions were made: 1.Incubation of mid-logarithmic E. coli cells with 5% of MAG or 2-DOG prior to drying enhanced preservation survival of bacteria.\",\n \"2.Incubation with more than 5% of nonmetabolizable sugar analogs had no additional effect on preservation survival.\",\n \"3.Accumulation of glucose in the cells had no protective effect against desiccation stress during preservation.\",\n \"EXAMPLE 6 Accumulation of MAG in the cells prior to drying enhances preservation of logarithmic E. coli cells.\",\n \"Two cell concentrates (harvested at mid-log growth stage) were prepared as described previously.\",\n \"To induce uptake of MAG, 0.5% glucose was added to the growth medium (“induced culture”).\",\n \"Both concentrates were incubated with 10% MAG at 37° C. for 30 minutes (loading).\",\n \"When loading was completed, one concentrate was diluted 100-fold in PBS buffer and incubated at 37° C. for an additional 30 minutes to induce expulsion of the pre-accumulated MAG.\",\n \"Both concentrates were preserved as described previously.\",\n \"Cell density in the concentrates was 4.7×108 cfu/ml.\",\n \"Preservation yield in loaded concentrate, which was not subsequently diluted, was 57% (2.7×108 cfu/ml).\",\n \"In contrast, in the concentrate, which was diluted to remove MAG pre-accumulated in the cells, preservation yield was only 3.8% (1.8×107 cfu/ml).\",\n \"Cells loaded with MAG were stable at room temperature for extended periods of time (i.e.\",\n \"stability after 12 days was 52%, 2.5×108 cfu/ml).\",\n \"Cells, which were depleted from pre-accumulated MAG by expulsion, and preserved at low yield, remained at 2.4% (1.1×107 cfu/ml).\",\n \"Based on the data of this example, the following conclusions were made: 1.Accumulation of non-metabolizable carbohydrate analogs, such as MAG, prior to drying enhances preservation survival of mid-log cells.\",\n \"2.Removal of the accumulated sugar by expulsion results in sensitivity to desiccation and low preservation yield.\",\n \"EXAMPLE 7 Mid-logarithmic culture was prepared as described previously.\",\n \"0.5% glucose was added to the growth medium (“induced cells”).\",\n \"The culture was concentrated 10-fold and the concentrates were incubated for 30 minutes with 10% MAG at different temperatures (0, 20, 40, and 60° C.).\",\n \"The mixtures were preserved as described previously.\",\n \"Cell densities (cfu/ml) in mid-logarithmic E. coli culture were 1.6×108 cfu/ml.\",\n \"Cell density in the concentrate was 1.9×109 cfu/ml.\",\n \"Preservation yield and stability of E. coli cells incubated with 10% MAG at different temperatures are shown in Table 8 and in FIGS.\",\n \"5 and 6.TABLE 8 Preservation Yield Stability (9 Days at RT°) Stability (16 Days at RT°) Sample % % % Description Cfu/ml Survival Cfu/ml Survival Cfu/ml Survival Loaded at 1.6 × 108 +/− 8.2 +/− 1.4 9.4 × 107 +/− 5.0 +/− 0.6 9.9 × 107 +/− 5.2 +/− 1.5 0° C. 2.7 × 107 1.2 × 107 2.8 × 107 Loaded at 2.7 × 108 +/− 14.1 +/− 2.6 1.6 × 108 +/− 8.7 +/− 0.6 1.4 × 108 +/− 7.3 +/− 1.7 20° C. 4.9 × 107 6.7 × 107 3.2 × 107 Loaded at 4.4 × 108 +/− 23.2 +/− 3.5 5.3 × 108 +/− 28.1 +/− 3.5 3.9 × 108 +/− 21.0 +/− 1.9 40° C. 6.6 × 107 6.6 × 107 3.7 × 107 Loaded at 1.4 × 104 +/− 0 1.2 × 104 +/− 0 1.3 × 104 +/− 0 60° C. 1.4 × 10−3 2.3 × 103 1.9 × 103 Logarithmic cells incubated with 10% MAG at 20° C. or at 40° C. preserved at higher yields and were more stable after 16 days of storage at room temperature compared to the cells incubated with MAG at 0° C. and at 60° C. (FIGS.\",\n \"5 and 6).\",\n \"Cells incubated with MAG at 40° C. preserved at higher yield than cells incubated with MAG at 20° C. and remained more stable during storage.\",\n \"Incubation at 60° C. resulted in severe loss in bacterial viability.\",\n \"Based on the data from this example, the following conclusions were reached: 1.Efficiency of loading of MAG into logarithmic E. coli cells is highly dependent on the incubation temperature.\",\n \"2.Loading of mid-log E. coli cells with MAG was the most effective when the cells were incubated at 20°-40° C., preferentially closer to 40° C. EXAMPLE 8 Effect of incubation time on accumulation of MAG in logarithmic E. coli cells.\",\n \"Cell concentrates were prepared as described previously (0.5% glucose was added to the growth medium) and incubated with 10% MAG at 37° C. for 0, 2.5, 5, 15, 30, and 60 minutes.\",\n \"The mixtures were preserved as described previously.\",\n \"Preservation yield and stability of mid-logarithmic E. coli cells incubated with 10% MAG for 0-60 minutes are shown in Table 9 and on FIG.\",\n \"7.TABLE 9 Preservation Yield Stability (7 days at RT) Stability (17 days at RT) Sample % % % Description Cfu/ml Survival Cfu/ml Survival Cfu/ml Survival Incubated 5.3 × 107 +/− 1.7 × 107 2.6 +/− 0.8 2.4 × 107 +/− 1.2 +/− 0.7 3.6 × 107 +/− 1.8 +/− 0.5 for 0 min 1.4 × 107 1.1 × 107 Incubated 1.2 × 108 +/− 2.1 × 107 5.8 +/− 1.1 9.6 × 107 +/− 4.9 +/− 0.8 5.9 × 107 +/− 2.9 +/− 0.6 for 2.5 min 1.5 × 107 1.2 × 107 Incubated 2.0 × 108 +/− 1.1 × 107 9.9 +/− 0.6 1.4 × 108 +/− 6.8 +/− 1.2 1.9 × 108 +/− 9.5 +/− 2.7 for 5 min 2.4 × 108 5.4 × 107 Incubated 3.4 × 108 +/− 5.6 × 107 16.8 +/− 2.8 2.1 × 108 +/− 10.4 +/− 0.4 2.4 × 108 +/− 11.8 +/− 2.6 for 15 min 8.3 × 106 5.2 × 107 Incubated 4.0 × 108 +/− 4.4 × 107 19.9 +/− 2.2 2.6 × 108 +/− 13.0 +/− 2.7 3.4 × 108 +/− 16.8 +/− 2.6 for 30 min 5.4 × 107 5.2 × 107 Incubated 4.4 × 108 +/− 6.1 × 107 22.2 +/− 3.0 3.3 × 108 +/− 16.3 +/− 1.9 3.9 × 108 +/− 19.3 +/− 3.9 for 60 min 3.7 × 107 7.7 × 107 Logarithmic cells incubated with MAG for 2.5-60 minutes preserved at higher yields than cells incubated with MAG for 0-2.5 minutes.\",\n \"Preservation survival of mid-logarithmic E. coli cells was directly proportional to the time of incubation with MAG (FIG.\",\n \"7).\",\n \"After 17 days of storage at RT, the cells incubated with MAG for at least 2.5 minutes were more stable than the cells incubated with MAG for less than 2.5 minutes.\",\n \"The following conclusions were reached, based on the data of this example: 1.Incubation with MAG for at least 2.5 minutes enhances preservation survival of mid-log E. coli cells.\",\n \"Loading for at least 5 minutes enhanced stability of the preserved cells.\",\n \"2.Preservation yield and stability of the preserved cells are directly proportional to the length of incubation with MAG.\",\n \"EXAMPLE 9 Effect of growth stage on accumulation of MAG in E. coli cells.\",\n \"E. coli cultures were grown in L-broth with 0.5% glucose as described previously and harvested at OD550=0.1 (early-log), 0.5 (mid-log), 1.0 (late-log) and 2.0 (stationary cells).\",\n \"The cells in each culture were concentrated and split in two aliquots.\",\n \"One aliquot of each concentrate was incubated with 10% MAG at 37° C. for 30 minutes (“loaded”).\",\n \"The second aliquot was not loaded (controls).\",\n \"Unloaded controls and loaded cells were preserved as described previously.\",\n \"Cell densities at different ODs were the following: 4.2×107 cfu/ml (OD550=0.1), 1.7×108 cfu/ml (OD550=0.5), 8.0×108 cfu/ml (OD550=1.0) and 1.3×109 cfu/ml (OD550=2.0).\",\n \"Preservation yield of E. coli harvested at different growth stages and loaded with 10% MAG is shown in Table 10 and on FIG.\",\n \"8.TABLE 10 Preservation Yield (Day 0) Stability (Day 7 at RT) Stability (Day 14) Sample % % % Description Cfu/ml Survival Cfu/ml Survival Cfu/ml Survival OD = 0.1, 1.6 × 107 +/− 10.9 +/− 3.3 8.9 × 106 +/− 6.1 +/− 1.0 1.1 × 107 +/− 7.0 +/− 1.9 control 4.8 × 106 1.4 × 106 2.8 × 106 OD = 0.1, 1.6 × 107 +/− 11.3 +/− 0.9 1.2 × 107 +/− 8.2 +/− 0.8 1.5 × 107 +/− 9.9 +/− 2.2 loaded 1.3 × 106 1.2 × 106 3.2 × 106 OD = 0.5, 2.6 × 108 +/− 10.7 +/− 0.6 2.5 × 108 +/− 10.0 +/− 1.3 1.8 × 108 +/− 7.3 +/− 0.3 control 1.5 × 107 3.3 × 107 8.3 × 105 OD = 0.5, 5.5 × 108 +/− 22.2 +/− 2.9 5.3 × 108 +/− 21.6 +/− 4.7 4.3 × 108 +/− 17.4 +/− 1.2 loaded 7.2 × 107 1.2 × 108 2.9 × 107 OD = 1.0, 1.3 × 109 +/− 15.1 +/− 3.7 1.4 × 109 +/− 16.2 +/− 2.8 1.6 × 109 +/− 18.4 +/− 4.6 control 3.3 × 108 2.5 × 108 4.0 × 108 OD = 1.0, 2.3 × 109 +/− 26.3 +/− 4.7 3.2 × 109 +/− 36.1 +/− 7.5 2.9 × 109 +/− 33.0 +/− 5.7 loaded 4.2 × 108 6.5 × 108 4.9 × 108 OD = 2.0, 3.5 × 109 +/− 49 +/− 5.4 4.1 × 109 +/− 57.5 +/− 6.8 4.3 × 109 +/− 60.3 +/− 11.5 control 3.8 × 108 4.8 × 108 8.2 × 108 OD = 2.0, 2.8 × 109 +/− 38.8 +/− 5.8 2.1 × 109 +/− 29.9 +/− 3.6 2.5 × 109 +/− 35.3 +/− 7.5 loaded 4.1 × 108 2.6 × 108 5.3 × 108 No significant difference in preservation survival between loaded and unloaded cells were found when the cells were harvested in early logarithmic stage (at OD005=0.1) before loading with 10% MAG.\",\n \"In contrast, the cells harvested in mid-log or late-log growth stage and loaded with MAG preserved at higher yield and were significantly more stable than unloaded controls.\",\n \"The stationary cells, loaded or unloaded, preserved at higher yield than loaded cells.\",\n \"High preservation survival in stationary cells could be attributed to the presence of intracellular protective compounds.\",\n \"The importance of the growth stage for efficient loading of MAG is clearly illustrated on FIG.\",\n \"8.Regardless of a mechanism of MAG accumulation, significant difference in preservation survival and stability between control and loaded cells was obtained only in early- and mid-logarithmic cells suggesting that the loading process was growth-stage dependent.\",\n \"Based on the data from this example, the following conclusions were reached: 1.Efficiency of loading the cells with MAG is highly dependent upon growth stage of bacteria.\",\n \"2.In vitro loading with MAG significantly increased preservation yield and stability in mid-log and late-log cells.\",\n \"3.Loading with MAG resulted in a decrease in preservation survival in stationary cells.\",\n \"4.Early-log cells preserved at lower yield and were less stable compared to the cells in later growth stage.\",\n \"5.Stationary cells were the most stable regardless of loading.\",\n \"EXAMPLE 10 Chemically competent E. coli XL10-Gold cells (1.5 L; Stratagene) in the form of a bacterial pellet were preserved using foam formation.\",\n \"The cells were concentrated 50-fold by resuspending the pellets in 30 ml of transformation buffer (Stratagene) and processed in the following manner.\",\n \"This example provides method steps which differ than those previously disclosed.\",\n \"Therefore, the methods of this example are included herein in their entirety.\",\n \"Methods (1) Cell density: To determine the cell density in the material, the cells (2×0.1 ml) were diluted in SM buffer (Stratagene) and plated in duplicate (0.1 ml) on L-agar plates.\",\n \"(2) Transformation of fresh cells: 100 l of cells+0.1 ng or 0.01 ng of pUC18 plasmid DNA (Stratagene) were mixed and incubated on ice for 30 minutes.\",\n \"When 30 minutes elapsed, the mixtures were transferred in a 42° C. water bath and subjected to a “heat shock” for 30 seconds.\",\n \"The mixtures were then incubated on ice for 2 minutes and 1 ml of NZY medium (Stratagene) was added.\",\n \"The mixtures were incubated at 37° C. for 60 minutes with shaking (200 rpm).\",\n \"The cells were plated on L-agar plates with 100 g/mi ampicilin (50, 100, 200, and 400 l aliquots of undiluted mixtures) for determination of transformation efficiency.\",\n \"The plates were incubated overnight at 37° C. (3) Freezing of the fresh cells: 10×0.15 ml of the original concentrate were aliquoted in Eppendorf tubes, mixed with 10% DMSO, and frozen at −80° C. Preservation yield and stability in the frozen cells was determined after storage at −80° C. for 3 and 7 days.\",\n \"(4) Preservation of electrocompetent cells by foam formation: The cells were split in two aliquots and mixed either with preservation solution “A” (22.5% 2:1 sucrose:raffinose) or with solution “B”(22.5% 4:1 sucrose:MAG) 43×0.5 ml of the mixture “A”was aliquoted in 43 sterile 5 ml vials.\",\n \"Similarly, 9×0.5 ml of the mixture “B” was aliquoted in 9 sterile 5 ml vials.\",\n \"Vials containing the two preservation mixtures were kept on ice until preservation.\",\n \"Bacterial survival in the preservation mixtures was determined by plating appropriate dilutions on L-agar plates.\",\n \"The plates were incubated overnight at 37° C. (5) Drying: Preservation mixtures were dried overnight by foam formation.\",\n \"Vials containing the mixture “A” were split in two groups and stored at 4° C. and at room temperature (RT).\",\n \"Vials containing the mixture “B” were stored at RT only.\",\n \"Stability of the preserved material was initially evaluated after 7 days of storage and will be monitored over time.\",\n \"(6) Determination of preservation yield: The preserved cells were rehydrated at RT with 0.5 ml of transformation buffer (STB) (Stratagene) or transformation buffer (UTB) (Universal Preservation Technologies).\",\n \"Stratagene's transformation buffer is available commercially.\",\n \"UTB buffer comprises: 10 mM MOPS, 75 mM calcium chloride, 10 mM rubidium chloride and 10% sucrose.\",\n \"Appropriate dilutions were plated on L-agar plates and the plates were incubated at 37° C. overnight.\",\n \"(7) Transformation of the preserved cells: The preserved cells were transformed in the same manner as the fresh cells.\",\n \"Results Chemically competent E. coli XL10-Gold cells (Stratagene) were preserved by the foam formation and by freezing at −80° C. The cells were preserved in two different solutions and rehydrated with two different buffers in order to determine possible effects of these parameters on the preservation yield.\",\n \"Preservation yield and stability of E. coli XL10-Gold cells were determined and presented in Table 11.TABLE 11 Preservation yield (Day 0) Stability (Day 7) Cells Cfu/ml % Cfu/ml % Fresh 1.0 × 1010 +/− 100 N/A N/A 2.6 × 108 Frozen 2.5 × 109 +/− 24.1 +/− 1.4* 2.5 × 109 +/− 1.1 × 108 24.4 +/− 1.0 1.5 × 108* Foam Formation Preserved PS “A”, 9.5 × 107 +/− 0.9 +/− 0.2 4° C. RT 4° C. RT UTB 2.1 × 107 3.9 × 107 +/− 1.0 × 107 3.5 × 107 0.38 +/− 0.34 +/− 0.10 +/− 2.3 × 107 0.22 PS “B”, 3.4 × 106 +/− 0.03 +/− 0.02 4° C. RT 4° C. RT UTB 1.9 × 106 N/A 1.8 × 105 N/A <0.01 +/− 9.3 × 104 PS “A”, 1.1 × 108 +/− 1.1 +/− 0.3 N/D N/D STB 2.8 × 107 PS “B”, 2.0 × 106 +/− 0.02 +/− 0.00 N/D N/D STB 2.5 × 105 PS, preservation solution UTB, transformation buffer (Universal Preservation Technologies) STB, transformation buffer (Stratagene) *Day 1 at −80° C. N/A, not applicable N/D, not determined Preservation yield of cells dried by foam formation were approximately 0.9% (Table 11).\",\n \"The yield was significantly lower than preservation yield of the frozen cells.\",\n \"Preservation yield in the cells preserved in solution “A” and rehydrated with UTB was comparable to that in the cells preserved in the same solution and rehydrated with STB.\",\n \"Similar findings were found for the cells preserved in solution “B”.\",\n \"However, overall preservation yield was significantly higher (>30-fold) when the cells were preserved in solution “A” compared to that obtained with solution “B” regardless of the rehydration buffer.\",\n \"After 7 days of storage at 4° C. and at RT, loss in viability of about ½ Log was observed in the material preserved in solution “A”.\",\n \"Also, a loss of about 1 Log was observed in the material preserved in solution “B”.\",\n \"There was no significant difference in stability of the material preserved in either solution at the two temperatures.\",\n \"No loss in viability was observed in frozen cells after 7 days at −80° C. Transformation efficiency of the preserved material is presented in Table 12.The cells were transformed as described in the methods section of this example.\",\n \"TABLE 12 Transformation Efficiency* Sample Description (Cfu/□g pUC18) Fresh 7.4 × 106 +/− 4.9 × 105 Frozen 3.1 × 106 +/− 3.7 × 105 Foam formation preserved (Day 0) PS “A”, UTB 7.6 × 106 +/− 1.2 × 106 PS “B”, UTB 8.0 × 105 +/− 1.2 × 105 PS “A”, STB 5.9 × 105 +/− 8.8 × 104 PS “B”, STB 3.8 × 104 +/− 1.7 × 104 Frozen (Day 4 at −80° C.) 1.1 × 106** Foam formation preserved (Day 7) PS “A”, UTB, stored at 4° C. 3.5 × 106 +/− 9.2 × 105 PS “A”, UTB, stored at RT 4.0 × 106 +/− 4.6 × 105 PS “B”, UTB, stored at RT 1.0 × 106 +/− 3.6 × 105 PS “A”, STB N/D PS “B”, STB N/D *Transformation efficiencies reported were obtained with 0.1 ng of pUC18.No significant difference was observed when 0.01 ng of pUC18 was used instead of 0.1 ng.\",\n \"**Only one sample was plated (0.4 ml) and a standard deviation could not be determined.\",\n \"N/D, not determined.\",\n \"Transformation efficiency in the cells preserved by foam formation was comparable to that in the fresh and frozen cells (Table 12).\",\n \"The cells preserved in solution “A” and rehydrated with transformation buffer (Universal Preservation Technologies) had the highest efficiency after drying.\",\n \"Storage at 4° C. or at RT did not compromise transformation efficiency of the preserved cells.\",\n \"There was no difference in the efficiency of the preserved cells stored for 7 days compared to the cells immediately after drying.\",\n \"The feasibility of preservation of chemically competent E. coli XL10-Gold cells was clearly demonstrated by the results discussed above.\",\n \"Differences in yield were observed for the cells preserved in the two different preservation solutions (“A” and “B”) evaluated in this experiment.\",\n \"There was no difference in the recovery of the cells preserved in solution “A” or “B” with respect to which rehydration solution was used (UTB or STB buffer).\",\n \"Overall, preservation survival in the cells dried by the foam formation was lower than for frozen cells (0.9-1.1% versus 24.1%).\",\n \"Chemically competent cells preserved in this example were harvested in a mid-logarithmic growth stage.\",\n \"Some initial loss in viability of the preserved cells was observed after 7 days of storage at either RT or 4° C. It is not unusual for cells preserved in logarithmic growth stage without desiccation protectants, like chemically competent cells used in the present example, to lose some stability after initial storage.\",\n \"Other protective solutions can be used to modulate stability of the preserved material in this example.\",\n \"Transformation efficiency in the preserved cells was comparable to that in the fresh and frozen cells.\",\n \"In contrast to preservation survival, where preservation solution was the factor critical to the recovery of the preserved cells, both preservation and rehydration solutions were critical parameters affecting the efficiency of the preserved cells.\",\n \"In addition, the cells rehydrated with buffer UTB transformed at efficiencies at least 10-fold higher than the cells rehydrated in buffer STB.\",\n \"Combination of preservation solution “A” and rehydration buffer (Universal Preservation Technologies) resulted in the cells which had the maximal transformation efficiency (7.6×106 cfu/g) after drying.\",\n \"Transformation efficiency in the preserved cells remained unchanged after storage for 7 days at 4° C. or at RT.\",\n \"In conclusion, chemically competent E. coli XL10-Gold cells were successfully preserved by foam formation and remained fully competent after preservation and a short-term storage.\",\n \"EXAMPLE 11 Electrocompetent E coli XL1Blue cells (3 L; Stratagene) in the form of a bacterial pellet were preserved by the foam formation process.\",\n \"The cells were concentrated 100-fold by resuspending the pellets in 30 ml concentrating solution (Universal Preservation Technologies) and processed in the following manner.\",\n \"This example provides method steps which differ than those previously disclosed.\",\n \"Therefore, the methods of this example are included in their entirety.\",\n \"Methods (1) Cell density: To determine the cell density in the material, the cells (2×0.1 ml) were diluted in SM buffer (Stratagene) and plated in duplicate (0.1 ml) on L-agar plates.\",\n \"(2) Electroporation of fresh cells: 40 l of cells+0.1 ng of pUC18 plasmid DNA (Stratagene) were mixed, transferred into 0.1 cm BioRad cuvette (Stratagene) and electroporated at E=1.7 kV, R=200 Ohm, C=25 F =4-5 msec).\",\n \"After the pulse, 1 ml of SOC medium (Stratagene) was added to the cells and the mixture was incubated at 37° C. for 60 minutes with shaking (250 rpm).\",\n \"The cells were plated on L-agar plates with no antibiotic (for determination of electroporation survival) and on L-agar plates with 100 g/ml ampicilin (for determination of electroporation efficiency).\",\n \"The plates were incubated overnight at 37° C. (3) Freezing of the fresh cells: 10×0.1 ml of the original concentrate was aliquoted in Eppendorf tubes, mixed with 10% DMSO, and frozen at −80° C. Preservation yield and stability in the frozen cells were determined after storage at −80° C. for 1 and 12 days.\",\n \"(4) Preservation of electrocompetent cells by foam formation: 3.6 ml of the cells were removed from the 30 ml of the material, centrifuged, and the pellet was resuspended in preservation solution #1 (Universal Preservation Technologies).\",\n \"The mixture was aliquoted in 16×1.2 ml vials (0.2 ml per vial).\",\n \"Similarly, 24.8 ml of the remaining cells were centrifuged and the pellet was resuspended in preservation solution #2 (Universal Preservation Technologies).\",\n \"The mixture was aliquoted in 16×1.2 ml vials (0.2 ml per vial) and in 43×5 ml vials (0.5 ml per vial).\",\n \"Vials containing the two preservation mixtures were kept on ice until preservation.\",\n \"Bacterial survival in preservation mixtures was determined by plating appropriate dilutions on L-agar plates.\",\n \"The plates were incubated overnight at 37° C. (5) Drying: Preservation mixtures were dried overnight by foam formation.\",\n \"Vials containing the preserved cells were stored at 4° C. and at room temperature (RT).\",\n \"The 1.2 ml vials were stored at 4° C. The 5 ml vials were divided in two groups and stored at RT and at 4° C. Stability of the preserved material was initially evaluated after 11 days of storage.\",\n \"(6) Determination of preservation yield: The preserved cells (1.2 ml vials, preservation solutions 1&2; 5 ml vials, preservation solution 2) were rehydrated at room temperature (RT) in solution “A” (Universal Preservation Technologies).\",\n \"0.5 ml of the solution was added to rehydrate the cells in 5 ml vials, 0.2 ml was used for rehydration of the cells in 1.2 ml vials.\",\n \"In addition, several 5 ml vials (material preserved in solution 2) were rehydrated with solution “B” (Universal Preservation Technologies).\",\n \"Appropriate dilutions were plated on L-agar plates and the plates were incubated at 37° C. overnight.\",\n \"(7) Electroporation of preserved cells: The preserved cells were electroporated in the same manner as the fresh cells.\",\n \"Results Electrocompetent E. coli XL1Blue cells (Stratagene) were preserved by foam formation and by freezing at −80° C. The cells were preserved in two different solutions, in two vial sizes, and rehydrated with two different solutions in order to determine a possible effect of these parameters on preservation yield.\",\n \"Preservation yield and stability of the E. coli XL1Blue cells were determined and presented in Table 13.TABLE 13 Stability (Day 12 frozen; Day 11 Foam Preservation yield (Day 0) Formation Preserved) Cells Cfu/ml % Cfu/ml % Fresh 6.5 × 109 +/− 1.8 × 109 100 N/A N/A Frozen N/D N/D 1.2 × 109 +/− 1.1 × 108 18.6 +/− 1.7 Foam Formation Preserved 1.2 ml vial, 2.6 × 108 +/− 1.7 × 107 4.0 +/− 0.3 N/D N/D PS#1, RS “A” 1.2 ml vial, 2.6 × 108 +/− 3.3 × 107 4.0 +/− 0.5 N/D N/D PS#2, RS “A” 5 ml vial, 3.5 × 108 +/− 5.3 × 107 5.3 +/− 0.8 N/D N/D PS#2, RS “A” N/D N/D 5 ml vial, 5.1 × 108 +/− 1.5 × 108 7.9 +/− 2.3 4° C. RT 4° C. RT PS#2, RS “B” 4.4 × 108 +/− 6.6 × 107 3.2 × 108 +/− 6.1 × 107 6.8 +/− 1.0 5.1 +/− 1.0 PS, preservation solution RS, rehydration solution N/A, not applicable N/D, not determined Preservation yield of foam formation dried cells was low, ranging from 4-8% (Table 13).\",\n \"No significant difference was found in preservation yield when the cells were preserved in two different solutions (PS#1 and PS#2) or in different size vials (1.2 ml or 5 ml) and rehydrated in solution “A” (Universal Preservation Technologies).\",\n \"Preservation yield was slightly higher when the cells preserved in 5 ml vials were rehydrated in solution “B” (Universal Preservation Technologies).\",\n \"The preserved cells were stable for 11 days at 4° C. and at RT.\",\n \"There was no significant difference in the stability of the preserved material at the two temperatures.\",\n \"Electroporation survival and efficiency of the preserved material are presented in Table 14.The cells were electroporated by the protocol as described previously in the methods section of this example.\",\n \"TABLE 14 Electroporation Electroporation Survival Efficiency Sample Description (%) (Cfu/□g pUC18) Fresh 19.3 +/2.1 8.7 × 107 +/− 5.6 × 106 Frozen (Day 1 at −80° C.) N/D 1.8 × 106 +/− 3.5 × 105 Frozen (Day 12 at −80° C.) 19.7 +/− 1.6 1.2 × 107 +/− 2.8 × 106 Foam Formation Preserved (Day 0) 1.2 ml vial, PS#1, RS “A” 18.1 +/− 5.9 2.5 × 105 +/− 1.3 × 105 1.2 ml vial, PS#2, RS “A” 24.1 +/− 7.8 9.5 × 105 +/− 5.2 × 105 5 ml vial, PS#2, RS “A” 16.0 +/6.5 2.0 × 105 +/− 1.6 × 105 5 ml vial, PS#2, RS “B” 5.6 +/0.4 7.3 × 105 +/− 3.3 × 105 Foam Formation Preserved (Day 11 at 4° C.) 5 ml vial, PS#2, RS “B” 8.9 +/− 2.0 1.0 × 106 +/− 1.7 × 105 Foam Formation Preserved (Day 11 at RT) 5 ml vial, PS#2, RS “B” 7.8 +/− 1.7 6.8 × 105 +/− 1.7 × 105 N/D, not determined Except for samples preserved in solution #2 (PS#2) in 5 ml vials and rehydrated in solution “B”, there was no significant difference in electroporation survival of the preserved E. coli XL1Blue cells at Day 0 compared to the fresh cells (Table 14).\",\n \"In addition, there was no significant difference in electroporation survival with respect to a vial size in the cells preserved in solution #1.The cells preserved in solution #2 in 5 ml vials and rehydrated in solution “B” survived electroporation at a lower rate than the cells rehydrated in solution “A”.\",\n \"Electroporation survival in the stored cells was determined for the sample preserved in solution #2 in 5 ml vials and rehydrated in solution “B”.\",\n \"There was no difference in electroporation survival after storage at 4° C. or at RT for 11 days compared to that at Day 0 (Table 2).\",\n \"Electroporation efficiency in the preserved cells was lower compared to that in the fresh or frozen cells (Table 14).\",\n \"When the cells were preserved in 5 ml vials and rehydrated in solution “B”, electroporation efficiency was higher than in the cells preserved under identical conditions and rehydrated in solution “A”.\",\n \"Storage at 4° C. or at RT did not compromise electroporation efficiency of the preserved cells.\",\n \"There was no difference in electroporation efficiency of the preserved cells stored for 11 days compared to the cells immediately after drying.\",\n \"Based on the data from this example, the following conclusions were made: 1.The feasibility of preservation of electrocompetent E. coli XL1Blue cells was confirmed in the experiment discussed in this example.\",\n \"2.With respect to formulation of preservation solution, no significant difference in the preservation yield was observed when the cells were preserved in two solutions evaluated in this experiment.\",\n \"3.The preserved electrocompetent cells were stable at 4° C. and at RT for 12 days.\",\n \"No significant difference in stability was observed at the two temperatures.\",\n \"4.Electroporation survival in the preserved cells was comparable to that in the fresh cells and was not compromised by storage at RT or at 4° C. 5.With the preservation and rehydration solutions used in this study, arcing was not a problem.\",\n \"Also, the formulations used for preservation and rehydration in this example resulted in an overall increase in the transformation efficiency of the preserved cells compared to the efficiency obtained in a similar experiment performed previously (7.3×105 cfu/ g pUC18 DNA compared to 5.0×104 cfu/ g).\",\n \"Electroporation efficiency in the preserved cells obtained in the experiment presented in this report (7.3×105 cfu/ g) was slightly lower than in the frozen cells (1.8×106 cfu/ g).\",\n \"In a separate study, we found that the concentration of sugars used in preservation and rehydration solutions has an inhibitory effect on electroporation efficiency when the cells were electroporated at a high voltage (1.7 kV).\",\n \"Therefore, to achieve the maximal efficiency of the cells, both preservation conditions (solutions to be used, drying protocols, rehydration solutions, etc.)\",\n \"and the conditions for electroporation (voltage, pulse duration, etc.)\",\n \"need to be optimized.\",\n \"Although the invention has been described in detail for the purposes of illustration, it is understood that such detail is solely for that purpose, and variations can be made therein by those skilled in the art without departing from the spirit and scope of the invention which is defined by the following claims.\",\n \"All references referred to above are hereby incorporated by reference.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10477014"}}},{"rowIdx":996,"cells":{"text":{"kind":"list like","value":[["Material for coating and product coated with the material","The invention relates to an iron based material for coating of surfaces which comprises a binding phase in the form of a corrosion dense alloy which contains at least 50% Fe, 12-30% Cr, maximally 5% Mn, 0-40% Ni, preferably 6-20% Ni and 0-7% Mo and 0-1% N and 6-20% Si, all stated as weight percent, which addition of Si lowers the liquidus temperature, that is the temperature at which the binding phase is completely melted.","The invention also aims at products of metal coated with the iron based material mentioned above."],["1-6.","(canceled) 7.An iron based material for coating surfaces, comprising a binding phase in the form of a corrosion dense alloy which contains at least 50% Fe, 12-30% Cr, maximally 5% Mn, 0-40% Ni, 0-7% Mo and 0-1% N and 6-20% Si, all stated as weight percent, which Si-addition lowers the liquidus temperature, that is the temperature at which the binding phase is completely melted, to a value below 1250° C., preferably below 1200° C. 8.An iron based material as defined by claim 7, wherein said 0-40% Ni is 6-20% Ni.","9.An iron based material according to claim 7, wherein the material comprises hard particles based on one or a mixture of two or more of oxides, nitrides, carbides and borides.","10.An iron based material as defined by claim 7, wherein one or a mixture of two or more of oxygen, carbon, nitrogen, and boron are added to the materials in such amounts that formation of one or a mixture of two or more of oxides, nitrides and carbides is formed.","11.An iron based material according to claim 7, wherein the binding phase apart from Si also contains one or a mixture of two or more of B, P, Hf, C, which elements also lower the liquidus temperature, at which the amount of B is 0-1,5%, while the amount of P is 0-15% and the amount of Hf is 0-15% and the amount of C 0-2%.","12.An iron based material according to claim 7, wherein the amount of Si in the binding phase is 7-16%.","13.An iron based material according to claim 12, wherein said 7-16% Si in the binding phase is 8-12%.","14.An iron based material according to claim 7, wherein the binding phase is produced by gas at least one of water atomisation of a melt, melt spinning of a melt, which melts have a composition as stated above, and by crushing an ingot with the composition as stated above."],["<SOH> BACKGROUND OF THE INVENTION <EOH>It is known earlier to make surfaces more wear resistant and corrosion resistant by coating them with an abrasion resistant material.","Such hard metal coatings usually consist of Ni- and Co-alloys, which form a softer binding phase and in addition contain hard particles such as oxides, carbides, borides and nitrides.","The hard particles may either be mixed with the binding phase or oxygen, carbon, nitrogen and/or boron are added in such amounts that these form oxides, carbides, nitrides and/or borides.","The abrasion resistant material is usually applied to the surface that should be treated in the form of a powder.","The powder is applied to the surface and the binding phase is melted.","Alternatively the material may already be melted when it is applied to the surface.","Coating material with a binding phase consisting of a Ni- or a Co-alloy may cause problems in that the coated surface in many instances is more noble than the parent metal which may give rise to galvanic corrosion.","These coating materials are also relatively expensive.","Today there are also coatings of stainless steel for certain applications.","The surfaces which shall be protected are coated with stainless steel, either by thermal spraying or by welding the stainless steel.","There are certain drawbacks.","If the stairdess steel is added by thermal spraying there is no tight layer.","If the stainless steel is welded, the material which is to be coated melts."],["<SOH> SUMMARY OF THE INVENTION <EOH>The present invention relates to a new material for coating surfaces, which material is proportionately inexpensive but offers the same possibility to obtain a tight layer which is well attached to the surface of the object which is coated as Ni- or Co-alloys.","The coating material is based on a corrosion dense material (stainless steel).","According to the invention the material comprises a binding phase of a corrosion dense alloy which contains at least 50% Fe, 12-30% Cr, maximally 5% Mn, 040% Ni, preferably 6-20% Ni, 0-7% Mo and 0-1% N and 6-20% Si, all stated as weight percent.","The addition of Si lowers the liquidus temperature, that is the temperature at which the binding phase is melted completely to a value below 1250° C., preferably below 1200° C. The coating material contains apart from iron and chromium, manganese and silicon in the amount given above.","The coating material may contain hard particles based on oxides, nitrides, carbides and/or mixtures of these.","Alternatively one may add oxygen, carbon, nitrogen and/or boron in such amounts that these elements form oxides, carbides, nitrides and/or borides.","The expression “iron based material” refers henceforth to the binding phase including the hard particles.","By coating a surface with an iron based material according to the invention one may obtain a corrosion dense surface layer on a surface that is not in itself corrosion resistant.","One may also obtain a wear resistant surface on an object manufactured from a less wear resistant material.","By a local coating of the iron based material one may, if so is desired, obtain a coating in certain points.","detailed-description description=\"Detailed Description\" end=\"lead\"?"],["CROSS-REFERENCE TO RELATED APPLICATIONS This application is entitled to the benefit of and incorporates by reference essential subject matter disclosed in International Application No.","PCT/SE02/00885 filed on May 6, 2002 and Swedish Patent Application No.","0101602-1 filed on May 7, 2001.FIELD OF THE INVENTION The present invention relates to an iron based material for coating of surfaces and products coated with the material.","BACKGROUND OF THE INVENTION It is known earlier to make surfaces more wear resistant and corrosion resistant by coating them with an abrasion resistant material.","Such hard metal coatings usually consist of Ni- and Co-alloys, which form a softer binding phase and in addition contain hard particles such as oxides, carbides, borides and nitrides.","The hard particles may either be mixed with the binding phase or oxygen, carbon, nitrogen and/or boron are added in such amounts that these form oxides, carbides, nitrides and/or borides.","The abrasion resistant material is usually applied to the surface that should be treated in the form of a powder.","The powder is applied to the surface and the binding phase is melted.","Alternatively the material may already be melted when it is applied to the surface.","Coating material with a binding phase consisting of a Ni- or a Co-alloy may cause problems in that the coated surface in many instances is more noble than the parent metal which may give rise to galvanic corrosion.","These coating materials are also relatively expensive.","Today there are also coatings of stainless steel for certain applications.","The surfaces which shall be protected are coated with stainless steel, either by thermal spraying or by welding the stainless steel.","There are certain drawbacks.","If the stairdess steel is added by thermal spraying there is no tight layer.","If the stainless steel is welded, the material which is to be coated melts.","SUMMARY OF THE INVENTION The present invention relates to a new material for coating surfaces, which material is proportionately inexpensive but offers the same possibility to obtain a tight layer which is well attached to the surface of the object which is coated as Ni- or Co-alloys.","The coating material is based on a corrosion dense material (stainless steel).","According to the invention the material comprises a binding phase of a corrosion dense alloy which contains at least 50% Fe, 12-30% Cr, maximally 5% Mn, 040% Ni, preferably 6-20% Ni, 0-7% Mo and 0-1% N and 6-20% Si, all stated as weight percent.","The addition of Si lowers the liquidus temperature, that is the temperature at which the binding phase is melted completely to a value below 1250° C., preferably below 1200° C. The coating material contains apart from iron and chromium, manganese and silicon in the amount given above.","The coating material may contain hard particles based on oxides, nitrides, carbides and/or mixtures of these.","Alternatively one may add oxygen, carbon, nitrogen and/or boron in such amounts that these elements form oxides, carbides, nitrides and/or borides.","The expression “iron based material” refers henceforth to the binding phase including the hard particles.","By coating a surface with an iron based material according to the invention one may obtain a corrosion dense surface layer on a surface that is not in itself corrosion resistant.","One may also obtain a wear resistant surface on an object manufactured from a less wear resistant material.","By a local coating of the iron based material one may, if so is desired, obtain a coating in certain points.","DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The iron based material contains apart from Si, B and/or P and/or Hf and/or C in the binding phase, which elements also lower the liquidus temperature.","The amount of B is 0-1,5%, while the amount of P is 0-15%, the amount of Hf is 0-15% and the amount of C is 0-2%.","The amount of Si which is found in the binding phase is 7-16%, preferably 8-12%.","The active, dissolved amount of Si ought to lie in this interval in order to obtain the desired melting point lowering effect.","The analysed amount of Si may be considerably higher, since Si may occur in the form of silicon carbides or silicon nitrides, be bound to oxygen or have been sintered in some other way.","The binding phase in the iron based coating material may if so is desired contain alloying elements such as Ti, W, Al, Cu, Nb, Ta, V and others.","Variations in composition may also be a consequence of small inevitable amounts of contaminating elements such as C, 0, N and S. The binding phase in the iron based coating materials produced by gas or water atomisation of a melt with the composition given above.","Alternatively the binding phase may be produced by melt spinning or crushing of an ingot with the composition as stated above.","The invention also includes products of metal coated with the iron based material of the kind described above.","The iron based material according to the invention may be applied to the surface, which shall be coated in manners known for other kinds of coatings by bringing the binding phase in the material to the melting point.","The iron based material is applied on the surface of the object which shall be coated, after which the object is heated in a furnace to a temperature where the binding phase of the material melts.","The iron based coating material may also be applied to the surface by thermal spraying.","The iron based materiel may also be applied to the surface in the shape of a powder, after which the binding phase is brought to melt by means of a laser.","In the table below there are shown examples of the composition of the binding phase in different coating materials, which are produced by melting in a small vacuum furnace.","The ingot thereafter solidified in the chill mould.","TABLE 1 Analysis of some experiment melts Melt Fe % Si % Mn % P % B % Cr % Mo % Ni 1 Bal 6 1 0 0 17 2.5 12 2 Bal 8 1 0 0 17 2.5 12 3 Bal 10 1 0 0 17 2.5 12 4 Bal 12 1 0 0 17 2.5 12 5 Bal 15 1 0 0 17 2.5 12 6 Bal 6 1 0 1.5 17 2.5 12 7 Bal 6 1 3 0 17 2.5 12 8 Bal 10 1.5 17 2.5 20 9 Bal 0 1 10.5 0 17 2.5 12 10 Bal 4 1 6 0 17 2.5 12 The expression Bal means that the remaining part of the material in the melt consists of Fe.","An iron based material of the kind that is proposed according to the invention is more advantageous from a cost viewpoint than the Ni- and Co-coatings that are found on the market today.","The now proposed coating material may be used for equipment within the food and pharmaceutical industry, where Ni-alloys are not allowed for coating of surfaces intended to be used in food connections.","The coating material may also find use for different kinds of surfaces which are subjected to hard wear as for example the screw/conveyor in a decanter centrifuge or other kinds of equipment which demand wear resistant surfaces or for example in grinding tools.","By coating the surface of, apparatuses which are used in a corrosive environment such as in the chemical industry or in salt water, as for example valves with the material according to the invention one may increase the corrosion resistance without having to manufacture the whole apparatus in a corrosion resistant material.","Also in other connections the coating material according to the invention may be used to obtain a more wear resistant/more corrosion resistant surface in equipment manufactured of a cheaper or more easily workable material.","The coating material according to the invention may also be used for coating of products where the composition of the binding phase closely corresponds to the composition of the material in the product."]],"string":"[\n [\n \"Material for coating and product coated with the material\",\n \"The invention relates to an iron based material for coating of surfaces which comprises a binding phase in the form of a corrosion dense alloy which contains at least 50% Fe, 12-30% Cr, maximally 5% Mn, 0-40% Ni, preferably 6-20% Ni and 0-7% Mo and 0-1% N and 6-20% Si, all stated as weight percent, which addition of Si lowers the liquidus temperature, that is the temperature at which the binding phase is completely melted.\",\n \"The invention also aims at products of metal coated with the iron based material mentioned above.\"\n ],\n [\n \"1-6.\",\n \"(canceled) 7.An iron based material for coating surfaces, comprising a binding phase in the form of a corrosion dense alloy which contains at least 50% Fe, 12-30% Cr, maximally 5% Mn, 0-40% Ni, 0-7% Mo and 0-1% N and 6-20% Si, all stated as weight percent, which Si-addition lowers the liquidus temperature, that is the temperature at which the binding phase is completely melted, to a value below 1250° C., preferably below 1200° C. 8.An iron based material as defined by claim 7, wherein said 0-40% Ni is 6-20% Ni.\",\n \"9.An iron based material according to claim 7, wherein the material comprises hard particles based on one or a mixture of two or more of oxides, nitrides, carbides and borides.\",\n \"10.An iron based material as defined by claim 7, wherein one or a mixture of two or more of oxygen, carbon, nitrogen, and boron are added to the materials in such amounts that formation of one or a mixture of two or more of oxides, nitrides and carbides is formed.\",\n \"11.An iron based material according to claim 7, wherein the binding phase apart from Si also contains one or a mixture of two or more of B, P, Hf, C, which elements also lower the liquidus temperature, at which the amount of B is 0-1,5%, while the amount of P is 0-15% and the amount of Hf is 0-15% and the amount of C 0-2%.\",\n \"12.An iron based material according to claim 7, wherein the amount of Si in the binding phase is 7-16%.\",\n \"13.An iron based material according to claim 12, wherein said 7-16% Si in the binding phase is 8-12%.\",\n \"14.An iron based material according to claim 7, wherein the binding phase is produced by gas at least one of water atomisation of a melt, melt spinning of a melt, which melts have a composition as stated above, and by crushing an ingot with the composition as stated above.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>It is known earlier to make surfaces more wear resistant and corrosion resistant by coating them with an abrasion resistant material.\",\n \"Such hard metal coatings usually consist of Ni- and Co-alloys, which form a softer binding phase and in addition contain hard particles such as oxides, carbides, borides and nitrides.\",\n \"The hard particles may either be mixed with the binding phase or oxygen, carbon, nitrogen and/or boron are added in such amounts that these form oxides, carbides, nitrides and/or borides.\",\n \"The abrasion resistant material is usually applied to the surface that should be treated in the form of a powder.\",\n \"The powder is applied to the surface and the binding phase is melted.\",\n \"Alternatively the material may already be melted when it is applied to the surface.\",\n \"Coating material with a binding phase consisting of a Ni- or a Co-alloy may cause problems in that the coated surface in many instances is more noble than the parent metal which may give rise to galvanic corrosion.\",\n \"These coating materials are also relatively expensive.\",\n \"Today there are also coatings of stainless steel for certain applications.\",\n \"The surfaces which shall be protected are coated with stainless steel, either by thermal spraying or by welding the stainless steel.\",\n \"There are certain drawbacks.\",\n \"If the stairdess steel is added by thermal spraying there is no tight layer.\",\n \"If the stainless steel is welded, the material which is to be coated melts.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The present invention relates to a new material for coating surfaces, which material is proportionately inexpensive but offers the same possibility to obtain a tight layer which is well attached to the surface of the object which is coated as Ni- or Co-alloys.\",\n \"The coating material is based on a corrosion dense material (stainless steel).\",\n \"According to the invention the material comprises a binding phase of a corrosion dense alloy which contains at least 50% Fe, 12-30% Cr, maximally 5% Mn, 040% Ni, preferably 6-20% Ni, 0-7% Mo and 0-1% N and 6-20% Si, all stated as weight percent.\",\n \"The addition of Si lowers the liquidus temperature, that is the temperature at which the binding phase is melted completely to a value below 1250° C., preferably below 1200° C. The coating material contains apart from iron and chromium, manganese and silicon in the amount given above.\",\n \"The coating material may contain hard particles based on oxides, nitrides, carbides and/or mixtures of these.\",\n \"Alternatively one may add oxygen, carbon, nitrogen and/or boron in such amounts that these elements form oxides, carbides, nitrides and/or borides.\",\n \"The expression “iron based material” refers henceforth to the binding phase including the hard particles.\",\n \"By coating a surface with an iron based material according to the invention one may obtain a corrosion dense surface layer on a surface that is not in itself corrosion resistant.\",\n \"One may also obtain a wear resistant surface on an object manufactured from a less wear resistant material.\",\n \"By a local coating of the iron based material one may, if so is desired, obtain a coating in certain points.\",\n \"detailed-description description=\\\"Detailed Description\\\" end=\\\"lead\\\"?\"\n ],\n [\n \"CROSS-REFERENCE TO RELATED APPLICATIONS This application is entitled to the benefit of and incorporates by reference essential subject matter disclosed in International Application No.\",\n \"PCT/SE02/00885 filed on May 6, 2002 and Swedish Patent Application No.\",\n \"0101602-1 filed on May 7, 2001.FIELD OF THE INVENTION The present invention relates to an iron based material for coating of surfaces and products coated with the material.\",\n \"BACKGROUND OF THE INVENTION It is known earlier to make surfaces more wear resistant and corrosion resistant by coating them with an abrasion resistant material.\",\n \"Such hard metal coatings usually consist of Ni- and Co-alloys, which form a softer binding phase and in addition contain hard particles such as oxides, carbides, borides and nitrides.\",\n \"The hard particles may either be mixed with the binding phase or oxygen, carbon, nitrogen and/or boron are added in such amounts that these form oxides, carbides, nitrides and/or borides.\",\n \"The abrasion resistant material is usually applied to the surface that should be treated in the form of a powder.\",\n \"The powder is applied to the surface and the binding phase is melted.\",\n \"Alternatively the material may already be melted when it is applied to the surface.\",\n \"Coating material with a binding phase consisting of a Ni- or a Co-alloy may cause problems in that the coated surface in many instances is more noble than the parent metal which may give rise to galvanic corrosion.\",\n \"These coating materials are also relatively expensive.\",\n \"Today there are also coatings of stainless steel for certain applications.\",\n \"The surfaces which shall be protected are coated with stainless steel, either by thermal spraying or by welding the stainless steel.\",\n \"There are certain drawbacks.\",\n \"If the stairdess steel is added by thermal spraying there is no tight layer.\",\n \"If the stainless steel is welded, the material which is to be coated melts.\",\n \"SUMMARY OF THE INVENTION The present invention relates to a new material for coating surfaces, which material is proportionately inexpensive but offers the same possibility to obtain a tight layer which is well attached to the surface of the object which is coated as Ni- or Co-alloys.\",\n \"The coating material is based on a corrosion dense material (stainless steel).\",\n \"According to the invention the material comprises a binding phase of a corrosion dense alloy which contains at least 50% Fe, 12-30% Cr, maximally 5% Mn, 040% Ni, preferably 6-20% Ni, 0-7% Mo and 0-1% N and 6-20% Si, all stated as weight percent.\",\n \"The addition of Si lowers the liquidus temperature, that is the temperature at which the binding phase is melted completely to a value below 1250° C., preferably below 1200° C. The coating material contains apart from iron and chromium, manganese and silicon in the amount given above.\",\n \"The coating material may contain hard particles based on oxides, nitrides, carbides and/or mixtures of these.\",\n \"Alternatively one may add oxygen, carbon, nitrogen and/or boron in such amounts that these elements form oxides, carbides, nitrides and/or borides.\",\n \"The expression “iron based material” refers henceforth to the binding phase including the hard particles.\",\n \"By coating a surface with an iron based material according to the invention one may obtain a corrosion dense surface layer on a surface that is not in itself corrosion resistant.\",\n \"One may also obtain a wear resistant surface on an object manufactured from a less wear resistant material.\",\n \"By a local coating of the iron based material one may, if so is desired, obtain a coating in certain points.\",\n \"DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The iron based material contains apart from Si, B and/or P and/or Hf and/or C in the binding phase, which elements also lower the liquidus temperature.\",\n \"The amount of B is 0-1,5%, while the amount of P is 0-15%, the amount of Hf is 0-15% and the amount of C is 0-2%.\",\n \"The amount of Si which is found in the binding phase is 7-16%, preferably 8-12%.\",\n \"The active, dissolved amount of Si ought to lie in this interval in order to obtain the desired melting point lowering effect.\",\n \"The analysed amount of Si may be considerably higher, since Si may occur in the form of silicon carbides or silicon nitrides, be bound to oxygen or have been sintered in some other way.\",\n \"The binding phase in the iron based coating material may if so is desired contain alloying elements such as Ti, W, Al, Cu, Nb, Ta, V and others.\",\n \"Variations in composition may also be a consequence of small inevitable amounts of contaminating elements such as C, 0, N and S. The binding phase in the iron based coating materials produced by gas or water atomisation of a melt with the composition given above.\",\n \"Alternatively the binding phase may be produced by melt spinning or crushing of an ingot with the composition as stated above.\",\n \"The invention also includes products of metal coated with the iron based material of the kind described above.\",\n \"The iron based material according to the invention may be applied to the surface, which shall be coated in manners known for other kinds of coatings by bringing the binding phase in the material to the melting point.\",\n \"The iron based material is applied on the surface of the object which shall be coated, after which the object is heated in a furnace to a temperature where the binding phase of the material melts.\",\n \"The iron based coating material may also be applied to the surface by thermal spraying.\",\n \"The iron based materiel may also be applied to the surface in the shape of a powder, after which the binding phase is brought to melt by means of a laser.\",\n \"In the table below there are shown examples of the composition of the binding phase in different coating materials, which are produced by melting in a small vacuum furnace.\",\n \"The ingot thereafter solidified in the chill mould.\",\n \"TABLE 1 Analysis of some experiment melts Melt Fe % Si % Mn % P % B % Cr % Mo % Ni 1 Bal 6 1 0 0 17 2.5 12 2 Bal 8 1 0 0 17 2.5 12 3 Bal 10 1 0 0 17 2.5 12 4 Bal 12 1 0 0 17 2.5 12 5 Bal 15 1 0 0 17 2.5 12 6 Bal 6 1 0 1.5 17 2.5 12 7 Bal 6 1 3 0 17 2.5 12 8 Bal 10 1.5 17 2.5 20 9 Bal 0 1 10.5 0 17 2.5 12 10 Bal 4 1 6 0 17 2.5 12 The expression Bal means that the remaining part of the material in the melt consists of Fe.\",\n \"An iron based material of the kind that is proposed according to the invention is more advantageous from a cost viewpoint than the Ni- and Co-coatings that are found on the market today.\",\n \"The now proposed coating material may be used for equipment within the food and pharmaceutical industry, where Ni-alloys are not allowed for coating of surfaces intended to be used in food connections.\",\n \"The coating material may also find use for different kinds of surfaces which are subjected to hard wear as for example the screw/conveyor in a decanter centrifuge or other kinds of equipment which demand wear resistant surfaces or for example in grinding tools.\",\n \"By coating the surface of, apparatuses which are used in a corrosive environment such as in the chemical industry or in salt water, as for example valves with the material according to the invention one may increase the corrosion resistance without having to manufacture the whole apparatus in a corrosion resistant material.\",\n \"Also in other connections the coating material according to the invention may be used to obtain a more wear resistant/more corrosion resistant surface in equipment manufactured of a cheaper or more easily workable material.\",\n \"The coating material according to the invention may also be used for coating of products where the composition of the binding phase closely corresponds to the composition of the material in the product.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10477020"}}},{"rowIdx":997,"cells":{"text":{"kind":"list like","value":[["Waste storage device","A container adapted for waste storage is provided.","The container has a mechanically operated sealing mechanism that allows a user to seal the waste storage film by depressing an actuator."],["1.A sealing mechanism for a waste storage container, said waste storage container having a body, a lid and a storage film cartridge adapted to be positioned in said body, said cartridge having a continuous length of storage film therein, said sealing mechanism comprising: an actuator operably connected to said lid and said cartridge, wherein actuation of said actuator opens said lid and seals said storage film.","2.The sealing mechanism of claim 1, wherein a first actuation of said actuator opens said lid and a second actuation of said actuator seals said storage film.","3.The sealing mechanism of claim 2, wherein said second actuation of said actuator closes said lid.","4.The sealing mechanism of claim 1, wherein said actuator is operably connected to said cartridge by a series of linkages.","5.The sealing mechanism of claim 5, wherein said body has a bottom, and wherein said actuator is positioned substantially near the bottom of said body.","6.The sealing mechanism of claim 5 wherein said actuator protrudes from said body.","7.The sealing mechanism of claim 1, wherein said actuator seals said storage film by rotating said cartridge.","8.The sealing mechanism of claim 7, further comprising a clutch operably connected to said actuator and said cartridge, said clutch allowing said cartridge to rotate in only one direction.","9.The sealing mechanism of claim 8, further comprising a rack gear operably connected to said actuator and said clutch, and a spring connected to said rack gear, wherein actuation of said actuator causes said rack gear to move in a first direction against the bias of said spring and release of said actuator causes said rack gear to move in a second direction under the bias of said spring to cause said clutch to rotate said cartridge.","10.The sealing mechanism of claim 9, further comprising a locking mechanism operably connected to said spring, wherein said actuator locks said spring in a compressed position upon a first actuation and said actuator unlocks said spring upon a second actuator.","11.The sealing mechanism of claim 5, wherein said actuator is pivotally connected to a first linkage and said first linkage is pivotally connected to a second linkage, said first linkage being operably connected to said lid to allow a user to open and close said lid and said second linkage being operably connected to said cartridge to allow a user to seal said storage film."],["<SOH> BACKGROUND OF THE INVENTION <EOH>1.Field of the Invention The present invention relates to a waste storage device or container.","More particularly, the present invention relates to a waste storage device having a mechanically operated sealing mechanism.","2.Description of the Prior Art Waste storage devices for disposal of waste, such as baby diapers, are known.","One such device is the Playtex Diaper Genie System®.","In the Diaper Genie System®, a film cartridge having a hole concentrically located therein, is positioned in the container body.","The film cartridge has a top surface from which a storage film extends and has a twist ring secured in the concentric hole of the cartridge.","The storage film extends from the film cartridge, over and then down through the twist ring, down through the hole in the film cartridge and into the container body.","When the user manually opens the lid assembly by hand, a diaper can be inserted through the twist ring and hole in the film cartridge, and in the storage film.","The twist ring is then manually rotated by the user's hand, which causes the film cartridge and storage film to rotate, tieing off and sealing the disposed diaper in the storage film.","The storage film is continuously fed from the film cartridge.","The device requires the user to rotate the twist ring by hand to seal the storage film.","In the present invention, the sealing operation is performed mechanically."],["<SOH> SUMMARY OF THE INVENTION <EOH>It is an object of the present invention to provide a waste storage device that has a mechanically operated sealing mechanism.","It is another object of the present invention to provide such a waste storage device that allows a user to operate the sealing mechanism with his or her foot.","The above objects and advantages of the present invention are provided by, and the present invention includes a sealing mechanism for a waste storage container that has a body, a lid and a storage film cartridge positioned in the body with the cartridge having a continuous length of storage film therein.","The sealing mechanism has an actuator operably connected to the lid and the cartridge, and actuation of the actuator opens the lid and seals the storage film.","Preferably, the sealing mechanism allows a first actuation of the actuator to open the lid and a second actuation of the actuator to seal the storage film.","More preferably, the sealing mechanism further comprises a rack gear operably connected to the actuator, a clutch, and a spring connected to the rack gear.","Actuation of the actuator causes the rack gear to move in a first direction against the bias of the spring and release of the actuator causes the rack gear to move in a second direction under the bias of the spring, which causes the clutch to rotate the cartridge.","In another embodiment of the present invention, a locking mechanism is operably connected to the spring and a first actuation of the actuator locks the spring in a compressed position while a second actuation of the actuator unlocks the spring.","Other and further objects, advantages and features of the present invention will be understood by reference to the following."],["BACKGROUND OF THE INVENTION 1.Field of the Invention The present invention relates to a waste storage device or container.","More particularly, the present invention relates to a waste storage device having a mechanically operated sealing mechanism.","2.Description of the Prior Art Waste storage devices for disposal of waste, such as baby diapers, are known.","One such device is the Playtex Diaper Genie System®.","In the Diaper Genie System®, a film cartridge having a hole concentrically located therein, is positioned in the container body.","The film cartridge has a top surface from which a storage film extends and has a twist ring secured in the concentric hole of the cartridge.","The storage film extends from the film cartridge, over and then down through the twist ring, down through the hole in the film cartridge and into the container body.","When the user manually opens the lid assembly by hand, a diaper can be inserted through the twist ring and hole in the film cartridge, and in the storage film.","The twist ring is then manually rotated by the user's hand, which causes the film cartridge and storage film to rotate, tieing off and sealing the disposed diaper in the storage film.","The storage film is continuously fed from the film cartridge.","The device requires the user to rotate the twist ring by hand to seal the storage film.","In the present invention, the sealing operation is performed mechanically.","SUMMARY OF THE INVENTION It is an object of the present invention to provide a waste storage device that has a mechanically operated sealing mechanism.","It is another object of the present invention to provide such a waste storage device that allows a user to operate the sealing mechanism with his or her foot.","The above objects and advantages of the present invention are provided by, and the present invention includes a sealing mechanism for a waste storage container that has a body, a lid and a storage film cartridge positioned in the body with the cartridge having a continuous length of storage film therein.","The sealing mechanism has an actuator operably connected to the lid and the cartridge, and actuation of the actuator opens the lid and seals the storage film.","Preferably, the sealing mechanism allows a first actuation of the actuator to open the lid and a second actuation of the actuator to seal the storage film.","More preferably, the sealing mechanism further comprises a rack gear operably connected to the actuator, a clutch, and a spring connected to the rack gear.","Actuation of the actuator causes the rack gear to move in a first direction against the bias of the spring and release of the actuator causes the rack gear to move in a second direction under the bias of the spring, which causes the clutch to rotate the cartridge.","In another embodiment of the present invention, a locking mechanism is operably connected to the spring and a first actuation of the actuator locks the spring in a compressed position while a second actuation of the actuator unlocks the spring.","Other and further objects, advantages and features of the present invention will be understood by reference to the following.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is a cross-sectional plan view of a waste storage device having the sealing mechanism of the present invention; FIG.","2 is a cross-sectional plan view of a portion of the sealing mechanism of FIG.","1; FIG.","3 is a perspective view of the waste storgae device of FIG.","1; FIG.","4 is a top view of the waste storage device of FIG.","1 without the lid; FIG.","5 is a cross-sectional plan view of a waste storage device having a second embodiment of the sealing mechanism of the present invention; FIG.","6 is a cross-sectional plan view of a portion of the sealing mechanism of FIG.","3; and FIG.","7 is a cross-sectional plan view of the waste storage device of FIG.","5 with the lid closed.","DETAILED DESCRIPTION OF THE INVENTION Referring to FIG.","1, there is shown a preferred embodiment of a waste storage device having the sealing mechanism of the present invention, generally represented by reference numeral 10.Waste storage device 10 has a body 100, a lid 120, a storage film cassette 130, a film ring 140 and a sealing mechanism 200.Body 100 has a base 105, an inner volume 110, front and rear portions 102, 104, and upper and lower portions 106, 108.Body 100 is substantially cylindrical in shape to maximize inner volume 110.However, alternative shapes for body 100 can also be used including rectangular or cubical.","Lower portion 108 of body 100 has an indentation or pedal housing 115 that will be described later in greater detail.","Preferably, pedal housing 115 is positioned along front portion 102.Lid 120 is substantially cylindrical in shape and has a diameter substantially the same size as the diameter of body 100 to provide a sealing engagement of the lid with the body along upper portion 106.If an alternative shape of body 100 is used, such as rectangular or cubical, then the corresponding shape would also be used for lid 120 to provide a sealing engagement of the lid and the body along upper portion 106.Lid 120 is pivotally connected to body 100 by lid hinge 125.Preferably, lid hinge 125 is positioned along rear portion 104.Lid 120 has a lid slot 127 formed therein.","Lid slot 127 is a u-shaped channel that is operably connected to sealing mechanism 200 to allow a user to open and close lid 120, which will be discussed later in greater detail.","Body 100 further has a cassette flange 117 with a center hole 118 and a circular wall 119.Flange 117 is circumferentially formed along the inner wall of body 100 in proximity to upper portion 106.Flange 117 engages with a clutch 270 of sealing mechanism 200, which will be discussed later in greater detail.","Center hole 118 provides a support for clutch 270 and for cassette 130, which is seated on the clutch.","Circular wall 119 extends substantially vertically upward from flange 117.As used herein, vertical refers to the direction between upper and lower portions 106, 108, which is substantially parallel with the sidewalls defining body 100.Horizontal refers to the direction between front and rear portions 102, 104, which is substantially perpendicular to the sidewalls defining body 100.Circular wall 119 has a diameter larger than the diameter of cassette 130.Circular wall 119 provides support for cassette 130 to prevent it from moving in a horizontal direction yet allowing it to rotate in center hole 118.Cassette 130 preferably has a cylindrical or ring-like shape with a center opening 132.Film 135 is a continuous sheet of film having a cylindrical shape that is stored in cassette 130.Cassette 130 also has a film ring 140 that is positioned in center opening 132.Film 135 is dispensed from the top of cassette 130 over the outside of film ring 140 and then down through the film ring and down through center opening 132 into inner volume 110.Film 135 is tied off or sealed at lower end 137 of the film in inner volume 110 to provide a storage volume 138.As will be discussed later in greater detail, in the preferred embodiment, after a diaper is inserted in storage volume 138, sealing mechanism 200 rotates cassette 130 to seal film upper end 139.Referring to FIGS.","1 through 4, sealing mechanism 200 of the present invention is shown.","Sealing mechanism 200 has an actuator 210, a foot pedal linkage 225, a rack linkage 230, a rack gear 260, a clutch 270 and a biasing mechanism 280.In the preferred embodiment, actuator 210 is a foot pedal and is angled downward for facilitating operation by a user's foot.","Pedal 210 protrudes out from body 100 through pedal orifice 116 and into pedal housing 115.Pedal housing 115 has a height that allows a user to slide his or her foot into the housing and onto pedal 210.Pedal 210 is pivotally connected to base 105 of body 100 at pedal pivot 215.This pivotal connection translates the user's downward force into an upward force for opening lid 120.Pedal 210 is pivotally connected to foot pedal linkage 225 at pedal linkage pivot 220.This pivotal connection translates the rotational movement of pedal 210 into a substantially vertical movement of pedal linkage 225.Pedal linkage 225 extends substantially vertically from lower portion 108 of body 100, through flange opening 227, through upper portion 106 of body 100, to lid 120.Pedal linkage 225 is pivotally secured to lid 120 such that an upward force along the pedal linkage causes the lid to rotate about lid hinge 125.In this embodiment, pedal linkage 225 is operably connected to lid 120 by lid slot 127 and lid tongue 250.Lid slot 127 is preferably a u-shaped channel formed in lid 120.Lid tongue 250 is preferably a cylindrical tongue that engages with lid slot 127.Preferably, lid tongue 250 has a cap to secure the tongue in lid slot 127.Pedal linkage 225 is also pivotally connected to rack linkage 230 at rack pivot 235.Rack linkage 230 is pivotally connected to rack gear 260 at rack pivot 240.The pivotal connections at rack pivots 235, 240 translate the substantially vertical movement of pedal linkage 225 into a substantially horizontal movement of rack linkage 230 and rack gear 260.Rack gear 260 is operably connected to clutch 270 so that a horizontal movement of the rack gear causes the clutch to rotate.","In this embodiment, clutch 270 has gears (not shown) that engage with gears 262 of rack gear 260.Rack gear 260 is further connected to a biasing mechanism 280 positioned on the opposite end of the rack gear from rack pivot 240.In this embodiment, biasing mechanism 280 is a coil spring that biases rack gear 260 toward rear portion 104 of body 100.Clutch 270 is removably connected to cartridge 130.Clutch 270 is a one-way clutch that only permits rotation of cartridge 130 in one direction.","Thus, rack gear 260 can horizontally move in two directions and clutch 270 can rotationally move in two directions.","However, cassette 130 can only rotate in one direction.","In operation, when a user depresses pedal 210, pedal linkage 225 moves in an upward direction.","This upward movement of pedal linkage 225 causes lid 120 to rotate about lid hinge 125 toward rear portion 104 of body 100.Thus, depressing pedal 210 causes the lid to open.","Simultaneously, the upward movement of pedal linkage 225 causes rack linkage 230 to move substantially horizontally toward front portion 102 of body 100.This causes rack gear 260 to move horizontally toward front portion 102.Rack gear 260 is operably connected to clutch 270, which causes the clutch to rotate in a first direction.","Cassette 130 remains stationary because clutch 270 is a one-way clutch and prevents the cassette from rotating in this first direction.","The horizontal movement of rack gear 260 towards front portion 102 also causes spring 280 to compress.","With lid 120 in an open position and storage volume 138 accessible to the user, waste storage, such as a diaper, can be inserted through film ring 140 into the storage volume.","The user then releases pedal 210 to commence the sealing operation.","The force of compressed spring 280 against rack gear 260 causes the rack gear to horizontally move back toward rear portion 104.The horizontal movement of rack gear 260 toward rear portion 104 causes clutch 270 to rotate in a second, opposite direction.","Cassette 130 rotates in this second direction.","The rotation of cassette 130 causes film 135 to tie off or seal near film upper end 139.Thus, release of pedal 210 causes sealing mechanism 200 to seal film 135.Simultaneously, the horizontal movement of rack gear 260 toward rear portion 104 causes rack linkage 230 to move toward rear portion 104.The horizontal movement of rack linkage 230 toward rear portion 104 causes movement of pedal linkage 225 in a substantially downward direction toward base 105.The movement of pedal linkage 225 in a substantially downward direction causes lid 120 to rotate about lid hinge 125 toward front portion 102.Thus, release of pedal 210 causes sealing mechanism to close lid 120.Referring to FIGS.","5 through 7, in an alternative embodiment, sealing mechanism 200 also has a locking mechanism 300.Locking mechanism 300 is operably connected to spring 280.When a user first actuates pedal 210 causing rack gear 260 to move toward front portion 102 and compressing spring 280, locking mechanism 300 is triggered and locks spring 280 in a compressed position.","Thus, lid 120 remains in an open position and a user can release his or her foot from pedal 210.When a user actuates pedal 210 for a second time causing rack gear 260 to move further toward front portion 102 and further compressing spring 280, locking mechanism 300 is again triggered and unlocks spring 280 from a compressed position.","Rack gear 260 is moved back toward rear portion 104 under the force of spring 280 causing rotation of clutch 270 and cassette 130, as well as downward movement of pedal linkage 225.Thus, film 135 is sealed by the rotation of cassette 130 and lid 120 is closed by the downward movement of pedal linkage 225.Pedal 210 returns to its upper position for the sealing operation to be repeated by a first and second downward depression of the pedal.","The present invention having thus been described with particular reference to the preferred forms thereof, it will be obvious that various changes and modifications may be made therein without departing from the spirit and scope of the present invention as defined in the appended claims."]],"string":"[\n [\n \"Waste storage device\",\n \"A container adapted for waste storage is provided.\",\n \"The container has a mechanically operated sealing mechanism that allows a user to seal the waste storage film by depressing an actuator.\"\n ],\n [\n \"1.A sealing mechanism for a waste storage container, said waste storage container having a body, a lid and a storage film cartridge adapted to be positioned in said body, said cartridge having a continuous length of storage film therein, said sealing mechanism comprising: an actuator operably connected to said lid and said cartridge, wherein actuation of said actuator opens said lid and seals said storage film.\",\n \"2.The sealing mechanism of claim 1, wherein a first actuation of said actuator opens said lid and a second actuation of said actuator seals said storage film.\",\n \"3.The sealing mechanism of claim 2, wherein said second actuation of said actuator closes said lid.\",\n \"4.The sealing mechanism of claim 1, wherein said actuator is operably connected to said cartridge by a series of linkages.\",\n \"5.The sealing mechanism of claim 5, wherein said body has a bottom, and wherein said actuator is positioned substantially near the bottom of said body.\",\n \"6.The sealing mechanism of claim 5 wherein said actuator protrudes from said body.\",\n \"7.The sealing mechanism of claim 1, wherein said actuator seals said storage film by rotating said cartridge.\",\n \"8.The sealing mechanism of claim 7, further comprising a clutch operably connected to said actuator and said cartridge, said clutch allowing said cartridge to rotate in only one direction.\",\n \"9.The sealing mechanism of claim 8, further comprising a rack gear operably connected to said actuator and said clutch, and a spring connected to said rack gear, wherein actuation of said actuator causes said rack gear to move in a first direction against the bias of said spring and release of said actuator causes said rack gear to move in a second direction under the bias of said spring to cause said clutch to rotate said cartridge.\",\n \"10.The sealing mechanism of claim 9, further comprising a locking mechanism operably connected to said spring, wherein said actuator locks said spring in a compressed position upon a first actuation and said actuator unlocks said spring upon a second actuator.\",\n \"11.The sealing mechanism of claim 5, wherein said actuator is pivotally connected to a first linkage and said first linkage is pivotally connected to a second linkage, said first linkage being operably connected to said lid to allow a user to open and close said lid and said second linkage being operably connected to said cartridge to allow a user to seal said storage film.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>1.Field of the Invention The present invention relates to a waste storage device or container.\",\n \"More particularly, the present invention relates to a waste storage device having a mechanically operated sealing mechanism.\",\n \"2.Description of the Prior Art Waste storage devices for disposal of waste, such as baby diapers, are known.\",\n \"One such device is the Playtex Diaper Genie System®.\",\n \"In the Diaper Genie System®, a film cartridge having a hole concentrically located therein, is positioned in the container body.\",\n \"The film cartridge has a top surface from which a storage film extends and has a twist ring secured in the concentric hole of the cartridge.\",\n \"The storage film extends from the film cartridge, over and then down through the twist ring, down through the hole in the film cartridge and into the container body.\",\n \"When the user manually opens the lid assembly by hand, a diaper can be inserted through the twist ring and hole in the film cartridge, and in the storage film.\",\n \"The twist ring is then manually rotated by the user's hand, which causes the film cartridge and storage film to rotate, tieing off and sealing the disposed diaper in the storage film.\",\n \"The storage film is continuously fed from the film cartridge.\",\n \"The device requires the user to rotate the twist ring by hand to seal the storage film.\",\n \"In the present invention, the sealing operation is performed mechanically.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>It is an object of the present invention to provide a waste storage device that has a mechanically operated sealing mechanism.\",\n \"It is another object of the present invention to provide such a waste storage device that allows a user to operate the sealing mechanism with his or her foot.\",\n \"The above objects and advantages of the present invention are provided by, and the present invention includes a sealing mechanism for a waste storage container that has a body, a lid and a storage film cartridge positioned in the body with the cartridge having a continuous length of storage film therein.\",\n \"The sealing mechanism has an actuator operably connected to the lid and the cartridge, and actuation of the actuator opens the lid and seals the storage film.\",\n \"Preferably, the sealing mechanism allows a first actuation of the actuator to open the lid and a second actuation of the actuator to seal the storage film.\",\n \"More preferably, the sealing mechanism further comprises a rack gear operably connected to the actuator, a clutch, and a spring connected to the rack gear.\",\n \"Actuation of the actuator causes the rack gear to move in a first direction against the bias of the spring and release of the actuator causes the rack gear to move in a second direction under the bias of the spring, which causes the clutch to rotate the cartridge.\",\n \"In another embodiment of the present invention, a locking mechanism is operably connected to the spring and a first actuation of the actuator locks the spring in a compressed position while a second actuation of the actuator unlocks the spring.\",\n \"Other and further objects, advantages and features of the present invention will be understood by reference to the following.\"\n ],\n [\n \"BACKGROUND OF THE INVENTION 1.Field of the Invention The present invention relates to a waste storage device or container.\",\n \"More particularly, the present invention relates to a waste storage device having a mechanically operated sealing mechanism.\",\n \"2.Description of the Prior Art Waste storage devices for disposal of waste, such as baby diapers, are known.\",\n \"One such device is the Playtex Diaper Genie System®.\",\n \"In the Diaper Genie System®, a film cartridge having a hole concentrically located therein, is positioned in the container body.\",\n \"The film cartridge has a top surface from which a storage film extends and has a twist ring secured in the concentric hole of the cartridge.\",\n \"The storage film extends from the film cartridge, over and then down through the twist ring, down through the hole in the film cartridge and into the container body.\",\n \"When the user manually opens the lid assembly by hand, a diaper can be inserted through the twist ring and hole in the film cartridge, and in the storage film.\",\n \"The twist ring is then manually rotated by the user's hand, which causes the film cartridge and storage film to rotate, tieing off and sealing the disposed diaper in the storage film.\",\n \"The storage film is continuously fed from the film cartridge.\",\n \"The device requires the user to rotate the twist ring by hand to seal the storage film.\",\n \"In the present invention, the sealing operation is performed mechanically.\",\n \"SUMMARY OF THE INVENTION It is an object of the present invention to provide a waste storage device that has a mechanically operated sealing mechanism.\",\n \"It is another object of the present invention to provide such a waste storage device that allows a user to operate the sealing mechanism with his or her foot.\",\n \"The above objects and advantages of the present invention are provided by, and the present invention includes a sealing mechanism for a waste storage container that has a body, a lid and a storage film cartridge positioned in the body with the cartridge having a continuous length of storage film therein.\",\n \"The sealing mechanism has an actuator operably connected to the lid and the cartridge, and actuation of the actuator opens the lid and seals the storage film.\",\n \"Preferably, the sealing mechanism allows a first actuation of the actuator to open the lid and a second actuation of the actuator to seal the storage film.\",\n \"More preferably, the sealing mechanism further comprises a rack gear operably connected to the actuator, a clutch, and a spring connected to the rack gear.\",\n \"Actuation of the actuator causes the rack gear to move in a first direction against the bias of the spring and release of the actuator causes the rack gear to move in a second direction under the bias of the spring, which causes the clutch to rotate the cartridge.\",\n \"In another embodiment of the present invention, a locking mechanism is operably connected to the spring and a first actuation of the actuator locks the spring in a compressed position while a second actuation of the actuator unlocks the spring.\",\n \"Other and further objects, advantages and features of the present invention will be understood by reference to the following.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is a cross-sectional plan view of a waste storage device having the sealing mechanism of the present invention; FIG.\",\n \"2 is a cross-sectional plan view of a portion of the sealing mechanism of FIG.\",\n \"1; FIG.\",\n \"3 is a perspective view of the waste storgae device of FIG.\",\n \"1; FIG.\",\n \"4 is a top view of the waste storage device of FIG.\",\n \"1 without the lid; FIG.\",\n \"5 is a cross-sectional plan view of a waste storage device having a second embodiment of the sealing mechanism of the present invention; FIG.\",\n \"6 is a cross-sectional plan view of a portion of the sealing mechanism of FIG.\",\n \"3; and FIG.\",\n \"7 is a cross-sectional plan view of the waste storage device of FIG.\",\n \"5 with the lid closed.\",\n \"DETAILED DESCRIPTION OF THE INVENTION Referring to FIG.\",\n \"1, there is shown a preferred embodiment of a waste storage device having the sealing mechanism of the present invention, generally represented by reference numeral 10.Waste storage device 10 has a body 100, a lid 120, a storage film cassette 130, a film ring 140 and a sealing mechanism 200.Body 100 has a base 105, an inner volume 110, front and rear portions 102, 104, and upper and lower portions 106, 108.Body 100 is substantially cylindrical in shape to maximize inner volume 110.However, alternative shapes for body 100 can also be used including rectangular or cubical.\",\n \"Lower portion 108 of body 100 has an indentation or pedal housing 115 that will be described later in greater detail.\",\n \"Preferably, pedal housing 115 is positioned along front portion 102.Lid 120 is substantially cylindrical in shape and has a diameter substantially the same size as the diameter of body 100 to provide a sealing engagement of the lid with the body along upper portion 106.If an alternative shape of body 100 is used, such as rectangular or cubical, then the corresponding shape would also be used for lid 120 to provide a sealing engagement of the lid and the body along upper portion 106.Lid 120 is pivotally connected to body 100 by lid hinge 125.Preferably, lid hinge 125 is positioned along rear portion 104.Lid 120 has a lid slot 127 formed therein.\",\n \"Lid slot 127 is a u-shaped channel that is operably connected to sealing mechanism 200 to allow a user to open and close lid 120, which will be discussed later in greater detail.\",\n \"Body 100 further has a cassette flange 117 with a center hole 118 and a circular wall 119.Flange 117 is circumferentially formed along the inner wall of body 100 in proximity to upper portion 106.Flange 117 engages with a clutch 270 of sealing mechanism 200, which will be discussed later in greater detail.\",\n \"Center hole 118 provides a support for clutch 270 and for cassette 130, which is seated on the clutch.\",\n \"Circular wall 119 extends substantially vertically upward from flange 117.As used herein, vertical refers to the direction between upper and lower portions 106, 108, which is substantially parallel with the sidewalls defining body 100.Horizontal refers to the direction between front and rear portions 102, 104, which is substantially perpendicular to the sidewalls defining body 100.Circular wall 119 has a diameter larger than the diameter of cassette 130.Circular wall 119 provides support for cassette 130 to prevent it from moving in a horizontal direction yet allowing it to rotate in center hole 118.Cassette 130 preferably has a cylindrical or ring-like shape with a center opening 132.Film 135 is a continuous sheet of film having a cylindrical shape that is stored in cassette 130.Cassette 130 also has a film ring 140 that is positioned in center opening 132.Film 135 is dispensed from the top of cassette 130 over the outside of film ring 140 and then down through the film ring and down through center opening 132 into inner volume 110.Film 135 is tied off or sealed at lower end 137 of the film in inner volume 110 to provide a storage volume 138.As will be discussed later in greater detail, in the preferred embodiment, after a diaper is inserted in storage volume 138, sealing mechanism 200 rotates cassette 130 to seal film upper end 139.Referring to FIGS.\",\n \"1 through 4, sealing mechanism 200 of the present invention is shown.\",\n \"Sealing mechanism 200 has an actuator 210, a foot pedal linkage 225, a rack linkage 230, a rack gear 260, a clutch 270 and a biasing mechanism 280.In the preferred embodiment, actuator 210 is a foot pedal and is angled downward for facilitating operation by a user's foot.\",\n \"Pedal 210 protrudes out from body 100 through pedal orifice 116 and into pedal housing 115.Pedal housing 115 has a height that allows a user to slide his or her foot into the housing and onto pedal 210.Pedal 210 is pivotally connected to base 105 of body 100 at pedal pivot 215.This pivotal connection translates the user's downward force into an upward force for opening lid 120.Pedal 210 is pivotally connected to foot pedal linkage 225 at pedal linkage pivot 220.This pivotal connection translates the rotational movement of pedal 210 into a substantially vertical movement of pedal linkage 225.Pedal linkage 225 extends substantially vertically from lower portion 108 of body 100, through flange opening 227, through upper portion 106 of body 100, to lid 120.Pedal linkage 225 is pivotally secured to lid 120 such that an upward force along the pedal linkage causes the lid to rotate about lid hinge 125.In this embodiment, pedal linkage 225 is operably connected to lid 120 by lid slot 127 and lid tongue 250.Lid slot 127 is preferably a u-shaped channel formed in lid 120.Lid tongue 250 is preferably a cylindrical tongue that engages with lid slot 127.Preferably, lid tongue 250 has a cap to secure the tongue in lid slot 127.Pedal linkage 225 is also pivotally connected to rack linkage 230 at rack pivot 235.Rack linkage 230 is pivotally connected to rack gear 260 at rack pivot 240.The pivotal connections at rack pivots 235, 240 translate the substantially vertical movement of pedal linkage 225 into a substantially horizontal movement of rack linkage 230 and rack gear 260.Rack gear 260 is operably connected to clutch 270 so that a horizontal movement of the rack gear causes the clutch to rotate.\",\n \"In this embodiment, clutch 270 has gears (not shown) that engage with gears 262 of rack gear 260.Rack gear 260 is further connected to a biasing mechanism 280 positioned on the opposite end of the rack gear from rack pivot 240.In this embodiment, biasing mechanism 280 is a coil spring that biases rack gear 260 toward rear portion 104 of body 100.Clutch 270 is removably connected to cartridge 130.Clutch 270 is a one-way clutch that only permits rotation of cartridge 130 in one direction.\",\n \"Thus, rack gear 260 can horizontally move in two directions and clutch 270 can rotationally move in two directions.\",\n \"However, cassette 130 can only rotate in one direction.\",\n \"In operation, when a user depresses pedal 210, pedal linkage 225 moves in an upward direction.\",\n \"This upward movement of pedal linkage 225 causes lid 120 to rotate about lid hinge 125 toward rear portion 104 of body 100.Thus, depressing pedal 210 causes the lid to open.\",\n \"Simultaneously, the upward movement of pedal linkage 225 causes rack linkage 230 to move substantially horizontally toward front portion 102 of body 100.This causes rack gear 260 to move horizontally toward front portion 102.Rack gear 260 is operably connected to clutch 270, which causes the clutch to rotate in a first direction.\",\n \"Cassette 130 remains stationary because clutch 270 is a one-way clutch and prevents the cassette from rotating in this first direction.\",\n \"The horizontal movement of rack gear 260 towards front portion 102 also causes spring 280 to compress.\",\n \"With lid 120 in an open position and storage volume 138 accessible to the user, waste storage, such as a diaper, can be inserted through film ring 140 into the storage volume.\",\n \"The user then releases pedal 210 to commence the sealing operation.\",\n \"The force of compressed spring 280 against rack gear 260 causes the rack gear to horizontally move back toward rear portion 104.The horizontal movement of rack gear 260 toward rear portion 104 causes clutch 270 to rotate in a second, opposite direction.\",\n \"Cassette 130 rotates in this second direction.\",\n \"The rotation of cassette 130 causes film 135 to tie off or seal near film upper end 139.Thus, release of pedal 210 causes sealing mechanism 200 to seal film 135.Simultaneously, the horizontal movement of rack gear 260 toward rear portion 104 causes rack linkage 230 to move toward rear portion 104.The horizontal movement of rack linkage 230 toward rear portion 104 causes movement of pedal linkage 225 in a substantially downward direction toward base 105.The movement of pedal linkage 225 in a substantially downward direction causes lid 120 to rotate about lid hinge 125 toward front portion 102.Thus, release of pedal 210 causes sealing mechanism to close lid 120.Referring to FIGS.\",\n \"5 through 7, in an alternative embodiment, sealing mechanism 200 also has a locking mechanism 300.Locking mechanism 300 is operably connected to spring 280.When a user first actuates pedal 210 causing rack gear 260 to move toward front portion 102 and compressing spring 280, locking mechanism 300 is triggered and locks spring 280 in a compressed position.\",\n \"Thus, lid 120 remains in an open position and a user can release his or her foot from pedal 210.When a user actuates pedal 210 for a second time causing rack gear 260 to move further toward front portion 102 and further compressing spring 280, locking mechanism 300 is again triggered and unlocks spring 280 from a compressed position.\",\n \"Rack gear 260 is moved back toward rear portion 104 under the force of spring 280 causing rotation of clutch 270 and cassette 130, as well as downward movement of pedal linkage 225.Thus, film 135 is sealed by the rotation of cassette 130 and lid 120 is closed by the downward movement of pedal linkage 225.Pedal 210 returns to its upper position for the sealing operation to be repeated by a first and second downward depression of the pedal.\",\n \"The present invention having thus been described with particular reference to the preferred forms thereof, it will be obvious that various changes and modifications may be made therein without departing from the spirit and scope of the present invention as defined in the appended claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10477041"}}},{"rowIdx":998,"cells":{"text":{"kind":"list like","value":[["Methods and compositions for controlling pests","The present invention is directed to pest-controlling compositions comprising as active ingredients one or more β-diones, particularly β-diketones and β-triketones, and to the use of these compositions inter alia for preventing, eradicating, destroying, repelling or mitigating harmful, annoying or undesired pests including insects, arachnids, helminths, molluscs, protozoa and viruses.","The present invention is further directed to processes of preparing β-diones by de novo synthesis or from natural sources such as volatile oil-bearing plants from families including Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae."],["1.-24.","(canceled) 25.A method for controlling harmful, annoying or undesired pests, other than plants, microbes, helminths and nematodes, said method comprising exposing said pests to a pest-controlling effective amount of a composition comprising a β-dione compound represented by the general formula (I) wherein A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C1, trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R&, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7; R6 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR9R10), NR9R10 or NR9OR7; R7 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R8 is selected from H, C1C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR11, SR11 or NR9R10; R9 and R10 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR12 or SR12; R11 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R12 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; X and Y are independently selected from oxygen, sulfur, —N—R4; and Q completes a 5-8-member saturated or unsaturated carbocyclic or heterocyclic ring.","26.The method of claim 25, wherein the compound is represented by the general formula (III) wherein X, Y and Z are each independently selected from oxygen, sulfur, —N—R4 or one of C═X, C═Y or C=Z is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; C, D, E and F are each independently selected from H, C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1; and R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2 (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R1)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7.27.The method of claim 26, wherein the compound is selected from flavesone (1-isobutyroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), isoleptospermone (1-isovaleroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), leptospermone (1-valeroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), papuanone (1-pentoyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), grandiflorone (1-(2-phenylethyl)-3,3,5,5-tetramethylcyclohexan-2,4,6-trione) and jensenone (1-valeroyl-3,5-dicarbonylcyclohexan-2,4,6-trione), including analogues and derivatives thereof.","28.The method of claim 26, wherein the compound has a structural formula selected from: 29.The method of claim 26, wherein the compound has a structural formula selected from: 30.The method of claim 26, wherein the compound has a structural formula selected from: 31.The method of claim 26, wherein the compound has a structural formula selected from: 32.The method of claim 25, wherein the compound is represented by the general formula (IV) wherein X and Y are each independently selected from oxygen, sulfur —N—R4 or one of C═X or C═Y is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; C, D, E and F are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2; and R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 cloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10, heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R1 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7.33.The method of claim 32, wherein the compound is selected from tasmanone (1-isobutroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), agglomerone (1-isobutroyl-4-methoxy-5,5-dimethylcyclohex-3-en-2,6-dione), lateriticone (1-valeroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), isolateriticone (1-isovaleroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione and platyphyllol (6,6-dimethyl-2-acetyl-5-methoxycyclohex-4-ene-1,3-dione), including analogues and derivatives thereof.","34.The method of claim 32, wherein the compound has a structural formula selected from: 35.The method of claim 25, wherein the compound is represented by the general formula (V) wherein X and Y are independently selected from oxygen, sulfur or —N—R4; and A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl C2-C10 alkenyl, aryl or heteroaryl; C, D, E, F, G and H are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; and R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7.36.The method of claim 35, wherein the compound is selected from are angustione (1-acetyl-3,5,5-trimethylcyclohex-2,6-dione), dehydroangustione (1-acetyl-3,5,5-trimethylcyclohex-3-en-2,6-dione) and xanthostemone (1-isobutroyl-5,5-dimethylcyclohex-3-en-2,6-dione), including their analogues and derivatives.","37.The method of claim 35, wherein the compound has a structural formula selected from: 38.The method of claim 35, wherein the compound has a structural formula selected from: 39.The method of claim 35, wherein the compound has a structural formula selected from: 40.The method of claim 25, wherein the compound is obtainable from a volatile oil-bearing organism.","41.The method of claim 40, wherein the compound is present in an essential oil derived from said volatile oil-bearing organism.","42.The method of claim 40, wherein the volatile oil-bearing organism is selected from volatile oil-bearing plants.","43.The method of claim 40, wherein the volatile oil-bearing organism is selected from plants from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.","44.The method of claim 40, wherein the volatile oil-bearing organism is selected from genera of the Myrtaceae family.","45.The method of claim 40, wherein the volatile oil-bearing organism belongs to a genus selected from Angophora, Austromyrtus, Backhousia, Baeckea, Callistemon, Corymbia, Darwinia, Eucalyptus, Kunzea, Leptospermum, Melaleuca, Syzygium and Xanthostemon 46.The method of claim 25, wherein the pest that is controlled is selected from insects, arachnids, helminths, molluscs, protozoa and viruses.","47.The method of claim 25, wherein the pest that is controlled is selected from insects, arachnids, helminths and molluscs.","48.The method of claim 25, wherein the compound is used in the form of, a pest-controlling composition which comprises from about 0.00005% to about 90% by weight of said compound.","49.-73.","(canceled) 74.A method for controlling harmful, annoying or undesired pests, other than plants and microbes, said method comprising exposing said pests to a pest-controlling effective amount of a composition comprising a β-dione compound represented by the general formula (I) wherein A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7; R6 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR9R10), NR9R10 or NR9OR7; R7 is selected from H, C1C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R8 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR11, SR11 or NR9R10; R9 and R10 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR12 or SR12; R11 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R12 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; X and Y are independently selected from oxygen, sulfur, —N—R4; and Q completes a 5-8-member saturated or unsaturated carbocyclic or heterocyclic ring, with the proviso that when B═H1, C1-C10, alkyl or aryl, A is not CR3NR4R5 or C(R3)2OR2.75.The method of claim 74, wherein the compound is represented by the general formula (III) wherein X, Y and Z are each independently selected from oxygen, sulfur, —N—R4 or one of C═X, C═Y or C=Z is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; C, D, E and F are each independently selected from H, C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1; and R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalky, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R1)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7, with the proviso that when B═H1, C1-C10, alkyl or aryl, A is not CR3NR4R5 or C(R3)2OR2.76.The method of claim 75, wherein the compound is selected from flavesone (1-isobutyroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), isoleptospermone (1-isovaleroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), leptospermone (1-valeroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), papuanone (1-pentoyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), grandiflorone (1-(2-phenylethyl)-3,3,5,5-tetramethylcyclohexan-2,4,6-trione) and jensenone (1-valeroyl-3,5-dicarbonylcyclohexan-2,4,6-trione), including analogues and derivatives thereof.","77.The method of claim 75, wherein the compound has a structural formula selected from: 78.The method of claim 75, wherein the compound has a structural formula selected from: 79.The method of claim 75, wherein the compound has a structural formula selected from: 80.The method of claim 75, wherein the compound has a structural formula selected from: 81.The method of claim 74, wherein the compound is represented by the general formula (IV) wherein X and Y are each independently selected from oxygen, sulfur —N—R4 or one of C═X or C═Y is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; C, D, E and F are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2; and R1 is selected from H, C1-C10 alkyl C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl.","C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy.","C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O2) or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7, with the proviso that when B═H1, C1-C10, alkyl or aryl, A is not CR3NR4R5 or C(R3)2OR2.82.The method of claim 81, wherein the compound is selected from tasmanone (1-isobutroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), agglomerone (1-isobutroyl-4-methoxy-5,5-dimethylcyclohex-3-en-2,6-dione), lateriticone (1-valeroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), isolateriticone (1-isovaleroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione and platyphyllol (6,6-dimethyl-2-acetyl-5-methoxycyclohex-4-ene-1,3-dione), including analogues and derivatives thereof.","83.The method of claim 81, wherein the compound has a structural formula selected from: 84.The method of claim 74, wherein the compound is represented by the general formula (V) wherein X and Y are independently selected from oxygen, sulfur or —N—R4; and A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; C, D, E, F, G and H are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; and R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, c2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, c3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7, with the proviso that when B═H1, C1-C10, alkyl or aryl, A is not CR3NR4R5 or C(R3)2OR2.85.The method of claim 84, wherein the compound is selected from are angustione (1-acetyl-3,5,5-trimethylcyclohex-2,6-dione), dehydroangustione (1-acetyl-3,5,5-trimethylcyclohex-3-en-2,6-dione) and xanthostemone (1-isobutroyl-5,5-dimethylcyclohex-3-en-2,6-dione), including their analogues and derivatives.","86.The method of claim 35, wherein the compound has a structural formula selected from: 87.The method of claim 84, wherein the compound has a structural formula selected from: 88.The method of claim 84, wherein the compound has a structural formula selected from: 89.The method of claim 74, wherein the compound is obtainable from a volatile oil-bearing organism.","90.The method of claim 89, wherein the compound is present in an essential oil derived from said volatile oil-bearing organism.","91.The method of claim 89, wherein the volatile oil-bearing organism is selected from volatile oil-bearing plants.","92.The method of claim 89, wherein the volatile oil-bearing organism is selected from plants from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.","93.The method of claim 89, wherein the volatile oil-bearing organism is selected from genera of the Myrtaceae family.","94.The method of claim 89, wherein the volatile oil-bearing organism belongs to a genus selected from Angophora, Austromyrtus, Backhousia, Baeckea, Callistemon, Corymbia, Darwinia, Eucalyptus, Kunzea, Leptospermum, Melaleuca, Syzygium and Xanthostemon 95.The method of claim 74, wherein the pest that is controlled is selected from insects, arachnids, helminths, molluscs, protozoa and viruses.","96.The method of claim 74, wherein the pest that is controlled is selected from insects, arachnids, helminths and molluscs.","97.The method of claim 74, wherein the compound is used in the form of a pest-controlling composition which comprises from about 0.00005% to about 90% by weight of said compound.","98.The method of claim 25, wherein Q completes the 5-8-member saturated or unsaturated carbocyclic or heterocyclic ring in which one or more members comprise —C(═X)—; and wherein Q is substituted with one or more substituents selected from C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1.99.The method of claim 74, wherein Q completes the 5-8-member saturated or unsaturated carbocyclic or heterocyclic ring in which one or more members comprise —C(═X)—; and wherein Q is substituted with one or more substituents selected from C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1."],["<SOH> BACKGROUND OF THE INVENTION <EOH>Triketones have been used for many years as herbicides for the control of undesired vegetation.","Herbicidal triketones have been described, for example, in EP-A-338992, EP-A-336898, U.S. Pat.","No.","4,869,748, EP-A-186118, EP-A-186119, EP-A-186120, U.S. Pat.","No.","4,202,840, U.S. Pat.","No.","4,695,673, U.S. Pat.","No.","4,780,127, U.S. Pat.","No.","4,921,526, U.S. Pat.","No.","5,006,150, U.S. Pat.","No.","5,545,607, U.S. Pat.","No.","5,925,795, U.S. Pat.","No.","5,990,046, U.S. Pat.","No.","6,218,579, EP-A-249150, EP-A-137963, EP-A-394889, EP-A-506907 or EP-B-135191.Examples of herbicidal triketones are inter alia Sulcotrione (MIKADO®) whose chemical designation is 2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexandione, 2-(4-methylsulfonyloxy-2-nitrobenzoyl)-4,4,6,6-tetramethyl-1,3-cyclohexane dione; 3-(4-methylsulfonyloxy-2-nitrobenzoyl)-bicyclo-[3,2,1]octane-2,4-dione; 3-(4-methylsulfonyl-2-nitrobenzoyl)-bicyclo-[3,2,1]octane-2,4-dione; 4-(4-chloro-2-nitrobenzoyl)-2,6,6-trimethyl-2H-1,2-oxazine-3,5(4H, 6H)-dione; 3-(4-methylthio-2-nitrobenzoyl)-bicyclo[3,2,1]octane-2,4-dione; 4-(2-nitro-4-trifluoromethoxybenzoyl)-2,6,6-trimethyl-2H-1,2-oxazine-3,5(4H, 6H)-dione."],["<SOH> SUMMARY OF THE INVENTION <EOH>The instant invention is predicated in part on the discovery that β-diones, particularly β-diketones and β-triketones, such as those obtainable from volatile oil-bearing plants including plants from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae, exhibit significant pesticidal, especially insecticidal, arachnicidal, helminthicidal and/or molluscicidal activity.","This discovery has been reduced to practice in novel pest-controlling compositions and methods for their preparation and use, as described hereinafter."],["FIELD OF THE INVENTION THIS INVENTION relates generally to methods and compositions for controlling pests.","More particularly, the present invention relates to pest-controlling compositions comprising as active ingredients one or more β-diones, particularly β-diketones and β-triketones, and to the use of these compositions inter alia for preventing, eradicating, destroying, repelling or mitigating harmful, annoying or undesired pests including insects, arachnids, helminths, molluscs, protozoa and viruses.","The present invention further relates to processes of preparing β-diones by de novo synthesis or from natural sources such as volatile oil-bearing plants from families including Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.","Bibliographic details of various publications referred to in this specification are collected at the end of the description.","BACKGROUND OF THE INVENTION Triketones have been used for many years as herbicides for the control of undesired vegetation.","Herbicidal triketones have been described, for example, in EP-A-338992, EP-A-336898, U.S. Pat.","No.","4,869,748, EP-A-186118, EP-A-186119, EP-A-186120, U.S. Pat.","No.","4,202,840, U.S. Pat.","No.","4,695,673, U.S. Pat.","No.","4,780,127, U.S. Pat.","No.","4,921,526, U.S. Pat.","No.","5,006,150, U.S. Pat.","No.","5,545,607, U.S. Pat.","No.","5,925,795, U.S. Pat.","No.","5,990,046, U.S. Pat.","No.","6,218,579, EP-A-249150, EP-A-137963, EP-A-394889, EP-A-506907 or EP-B-135191.Examples of herbicidal triketones are inter alia Sulcotrione (MIKADO®) whose chemical designation is 2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexandione, 2-(4-methylsulfonyloxy-2-nitrobenzoyl)-4,4,6,6-tetramethyl-1,3-cyclohexane dione; 3-(4-methylsulfonyloxy-2-nitrobenzoyl)-bicyclo-[3,2,1]octane-2,4-dione; 3-(4-methylsulfonyl-2-nitrobenzoyl)-bicyclo-[3,2,1]octane-2,4-dione; 4-(4-chloro-2-nitrobenzoyl)-2,6,6-trimethyl-2H-1,2-oxazine-3,5(4H, 6H)-dione; 3-(4-methylthio-2-nitrobenzoyl)-bicyclo[3,2,1]octane-2,4-dione; 4-(2-nitro-4-trifluoromethoxybenzoyl)-2,6,6-trimethyl-2H-1,2-oxazine-3,5(4H, 6H)-dione.","SUMMARY OF THE INVENTION The instant invention is predicated in part on the discovery that β-diones, particularly β-diketones and β-triketones, such as those obtainable from volatile oil-bearing plants including plants from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae, exhibit significant pesticidal, especially insecticidal, arachnicidal, helminthicidal and/or molluscicidal activity.","This discovery has been reduced to practice in novel pest-controlling compositions and methods for their preparation and use, as described hereinafter.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 shows the structures relating to the major constituents of the published Myrtaceae essential oils.","FIG.","2 is a representation of a GC-MS trace of E. cloeziana oil.","FIG.","3 is a tabular and graphical representation showing 1H NMR data recorded on a fraction (F4) obtained from silica gel chromatography of E. cloeziana oil and the structure of the major and minor isomers of the compound deduced from these data.","FIG.","4 is a diagrammatic representation showing various tautomeric forms of an isolated β-triketone compound in solution (CDCl3).","DETAILED DESCRIPTION OF THE INVENTION One aspect of the present invention contemplates the use of a β-dione compound, particularly a β-diketone or a β-triketone compound, in the preparation of a composition for controlling harmful, annoying or undesired pests, said compound being represented by the general formula (I) wherein A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR5, (C—N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; R1 is selected from H, C1-C10 alkyl C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7; R6 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR9R10), NR9R10 or NR9OR7; R7 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R8 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR11, SR11 or NR9R10; R9 and R10 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR12 or SR12; R11 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R12 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; X and Y are independently selected from oxygen, sulfur, —N—R4; and Q completes a 5-8-member saturated or unsaturated carbocyclic or heterocyclic ring in which optionally one or more members comprise —C(═X)—; and wherein Q is, optionally substituted with one or more substituents selected from C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1.Heterocyclic systems can be optionally attached to a moiety other than those set forth above via a carbon atom or a heteroatom of R1 to R11.Preferred compounds represented by formula (1) are β-diketones and especially preferred are β-triketones.","As used herein, the term “alkyl” refers to linear or branched chains.","The term “haloalkyl” refers to an alkyl group substituted by at least one halogen.","Similarly the term “haloalkoxy” refers to an alkoxy group substituted by at least one halogen.","As used herein the term “halogen” refers to fluorine, chlorine, bromine and iodine.","As used herein the term “aryl” refers to aromatic carbocyclic ring systems such as phenyl or naphthyl, anthracenyl, especially phenyl.","Suitably, aryl is C6-C14 with mono, di, tri, tetra and penta substitution containing OR2, F, Cl, Br, I, NO2, CF3, COR1, NR4R5, SO2R2, SR2.As used herein the terms “heterocycle”, “heterocyclic”, “heterocyclic systems” and the like refer to a saturated, unsaturated, or aromatic carbocyclic group having a single ring, multiple fused rings (for example, bicyclic, tricyclic, or other similar bridged ring systems or substituents), or multiple condensed rings, and having at least one heteroatom such as nitrogen, oxygen, or sulfur within at least one of the rings.","This term also includes “heteroaryl” which refers to a heterocycle in which at least one ring is aromatic.","Any heterocyclic or heteroaryl group can be unsubstituted or optionally substituted with one or more groups, as defined above.","Further, bi- or tricyclic heteroaryl moieties may comprise at least one ring, which is either completely, or partially, saturated.","Suitable heteroaryl moieties include, but are not limited to oxazolyl, thiazaoyl, thienyl, furyl, 1-isobenzofuranyl, 2H-pyrrolyl, N-pyrrolyl, imidazolyl, pyrazolyl, isothiazolyl, isooxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyradazinyl, indolizinyl, isoindolyl, indoyl, indolyl, purinyl, phthalazinyl.","The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.","By way of example, “an element” means one element or more than one element.","A preferred carbocyclic ring formed by Q is an optionally substituted cyclohexanedione.","A preferred subgroup of compounds of formula (1) is represented by formula (II) Such compounds may exist in a number of tautomeric forms.","For example, in the case wherein X and Y are each oxygen, and B is hydrogen, then the compounds of formula II may exist as one or more of the structural formulae shown below.","It is intended that all such tautomeric structures are included within the scope of the present invention.","It should also be appreciated that some of the compounds of formula (I) are capable of existing as different geometric isomers and diastereomers.","The invention thus includes both the individual isomers and mixtures of such isomers.","Another preferred subgroup of compounds of formula (I) is represented by formula (II) wherein X, Y and Z are each independently selected from oxygen, sulfur, —N—R4 or one of C═X, C═Y or C=Z is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is as defined above; C, D, E and F are each independently selected from H, C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1; and R1, R2, R2, R4 and R5 are as defined above.","Preferred β-diones represented by formula (III) are flavesone (1-isobutyroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), isoleptospermone (1-isovaleroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), leptospermone (1-valeroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), papuanone (1-pentoyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), grandiflorone (1-(2-phenylethyl)-3,3,5,5-tetramethylcyclohexan-2,4,6-trione) and jensenone (1-valeroyl-3,5-dicarbonylcyclohexan-2,4,6-trione), including analogues and derivatives thereof.","By way of example, flavesone analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of flavesone is shown for comparative purposes: Non-limiting examples of isoleptospermone analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of isoleptospermone is shown for comparative purposes: Non-limiting examples of leptospermone analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of leptospermone is shown for comparative purposes: Non-limiting examples of jensenone analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of jensenone is shown for comparative purposes: Another preferred subgroup of compounds of formula (1) is represented by formula (IV) wherein X and Y are each independently selected from oxygen, sulfur —N—R4 or one of C═X or C—Y is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, N4, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is as defined above; C, D, E and F are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2; and R1, R2, R2, R4 and R5 are as defined above.","Preferred β-diones represented by formula (IV) are tasmanone (1-isobutroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), agglomerone (1-isobutroyl-4-methoxy-5,5-dimethylcyclohex-3-en-2,6-dione), lateriticone (1-valeroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), isolateriticone (1-isovaleroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione and platyphyllol (6,6-dimethyl-2-acetyl-5-methoxycyclohex-4-ene-1,3-dione), including analogues and derivatives thereof.","Non-limiting examples of tasmanone analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of tasmanone is shown for comparative purposes: Another preferred subgroup of compounds of formula (I) is represented by formula (V) wherein X and Y are independently selected from oxygen, sulfur or —N—R4; and A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is as defined above; C, D, E, F, G and H are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; and R1, R2, R2, R4 and R5 are as defined above.","More specifically unsaturation, epoxides and thioexpoxides may exist at positions designated by H (or G) connected to F (or E) or F (or E) connected to C (or D).","A four-membered ring forming a part of a bicyclic structure may exist at positions designated by H (or G) connected to C (or D).","Preferred β-diones represented by formula (V) are angustione (1-acetyl-3,5,5-trimethylcyclohex-2,6-dione), dehydroangustione (1-acetyl-3,5,5-trimethylcyclohex-3-en-2,6-dione) and xanthostemone (1-isobutroyl-5,5-dimethylcyclohex-3-en-2,6-dione), including their analogues and derivatives.","By way of example, angustione analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of angustione is shown for comparative purposes: Non-limiting examples of dehydroangustione analogues include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of dehydroangustione is shown for comparative purposes: Non-limiting examples of xanthostemone analogues include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of xanthostemone is shown for comparative purposes: Derivatives of the above compounds include, but are not restricted to, ethoxylate derivatives, propoxylate derivatives, hydrates, aldehyde derivatives, ester derivatives, ether derivatives, alcohol derivatives, phenol derivatives, amine derivatives, other biologically or chemically equivalent substances, and any combination of two or more of the foregoing.","Similarly effective as pesticides are salts of the above compounds, including mono-valent salts (e.g., sodium, potassium) and di-valent metal salts (e.g.","calcium, magnesium, iron or copper) and ammonium salts (e.g., isopropyl ammonium, trialkyl and tetraalkylammonium salts).","The compounds according to any one of formulae (I)-(V) can be prepared according to methods analogous to those known in the art for the preparation of β-diones.","Exemplary methods are disclosed for example in EP-A-338992, EP-A-336898, U.S. Pat.","No.","4,202,840, U.S. Pat.","No.","4,869,748, EP-A-186118, EP-A-186119, EP-A-186120, U.S. Pat.","No.","4,695,673, U.S. Pat.","No.","4,780,127, U.S. Pat.","No.","4,921,526, U.S. Pat.","No.","5,006,150, U.S. Pat.","No.","5,545,607, U.S. Pat.","No.","5,925,795, U.S. Pat.","No.","5,990,046, U.S. Pat.","No.","6,218,579, EP-A-249150, EP-A-137963, EP-A-394889 EP-A-506907 or EP-B-135191.More particularly, compounds according to formulae (III)-(V) can be synthesised using the representative procedures outlined below.","For compounds according to formula (III), 1,3,5-trihydroxybenzene 1 is reacted with CH3CN in the presence of zinc chloride and hydrochloric acid, according to A. H. Blatt (1943, Org.","Synth.","Col. II, 522-523), affording 1-acetyl-2,4,6-trihydroxybenzene 2 (phloroacetophenone) (R=Me) (Scheme 1).","R groups other than methyl are depicted above.","Reaction of 1-acetyl-2,4,6-trihydroxybenzene 2 affording 1-acetyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione 3 is a representative procedure for all compounds according to formula (III) (R. A.","Gray et al., U.S. Pat.","No.","4,202,840) (Scheme 2).","Anhydrous MeI (6eq) is slowly added, at room temperature under an atmosphere of nitrogen, to a mechanically stirred solution of 1-acetyl-2,4,6-trihydroxybenzene 2 (1 eq) and sodium ethoxide (6eq) in anhydrous methanol.","The mixture is refluxed for 4 hours.","On cooling the mixture is concentrated under vacuo, providing a residue, which is diluted with water and acidified with 2 M hydrochloric acid.","Diethylether extracts are washed with saturated sodium sulfite solution, water and then dried (Na2SO4).","Evaporation of the diethylether provides the desired product 3.For mono, di, tri and tetra B, C, D, E, F substitution patterns, reactions between one and seven mole equivalents of R—I and sodium ethoxide are used.","Lawasson's reagent is used for conversion of oxygen into sulfur groups and sodium borohydride or sodium cyanoborohydride is used to reduce ketone, thioketone and imino groups.","When additional carbonyl groups are introduced into the cyclohexane ring system the procedure of Crow is utilised (M. L. Bolte et al., 1985, Agric.","Biol.","Chem., 49, 761).","Compounds of formula (IV) can be prepared according to a first representative procedure, as follows: 3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 4 (1 mole eq), prepared according to Herzig (J. Herzig, and F. Wenzel, Monatsh, 1903, 24, 101), is dissolved in anhydrous diethylether and hexamethylphosphoramide (solvent ratio, 20:1 respectively) under an atmosphere of nitrogen.","The mixture is cooled to 0° C. and lithium hydride (1.1 mole eq) (60% in mineral oil) is added in portions.","After addition the mixture is stirred for a further 10 mins before the addition of benzoyl cyanide 5 [R—CO—CN, R is depicted above] (1.1 mole eq).","The mixture is allowed to warm to room temperature over 12 h at which time the reaction is quenched with water and partitioned.","The ether layer is dried (Na2SO4) and evaporated affording crude 1-benzoyl-3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 6 which is purified by SiO2 column chromatography (hexane/ethyl acetate, gradient) (Scheme 3).","Alternatively, compounds of formula (IV) can be prepared according to a second representative procedure, as follows: 3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 4 (1 mole eq) (commercially available) and benzoyl cyanide are dissolved in anhydrous dichloromethane and cooled to 0° C. under an atmosphere of nitrogen.","To the cooled solution is added anhydrous finely powdered zinc chloride (1.1 mole eq.)","followed by slow addition of triethylamine (1.2 mole eq).","The reaction mixture is stirred at room temperature for 5-6 h and then poured into 2 M hydrochloric acid.","The mixture is partitioned and the dichloromethane layer is washed with 5% sodium carbonate.","The aqueous carbonate phase is then acidified with hydrochloric acid and extracted with methylene chloride and dried (Na2SO4).","The solvent is removed and the residue subjected to SiO2 column chromatography (hexane/ethyl acetate) affording 1-benzoyl-3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 6 (W. J. Michaely and G. W. Kraatz, EP-B-135191).","Compounds of formula (V) can be prepared according to a first representative procedure, as follows: 4,4-dimethylcyclohexane-1,3-dione 7 (1 mole eq) (commercially available) is dissolved in anhydrous diethylether and hexamethylphosphoramide (solvent ratio, 20:1 respectively) under an atmosphere of nitrogen.","The mixture is cooled to 0° C. and lithium diisopropylamide (2.1 mole eq) is added dropwise over 40 mins.","The mixture is then stirred for a further 10 mins before the addition of methyl iodide (1 mole eq).","The mixture is stirred for 12 h and then benzoyl cyanide (2 mole eq) is added and the mixture stirred for a further 24 h. The reaction was quenched with water and the ether layer partitioned and dried (Na2SO4).","The solvent was removed and the residue subjected to SiO2 column chromatography (hexane/ethyl acetate) affording 1-benzoyl-3,3,5-trimethylcyclohexan-2,6-dione 8 (Scheme 4).","Alternatively, the compounds of formula (V) can be prepared according to a second representative procedure, as follows: 4,4-dimethyl-1,3-cyclohexanedione 7 (1 mole eq) (commercially available) and benzoyl cyanide are dissolved in anhydrous dichloromethane and cooled to 0° C. under an atmosphere of nitrogen.","To the cooled solution is added anhydrous finely powdered zinc chloride (1.1 mole eq) followed by slow addition of triethylamine (1.2 mole eq).","The reaction mixture is stirred at room temperature for 5-6 h and then poured into 2 M hydrochloric acid.","The mixture is partitioned and the dichloromethane layer washed with 5% sodium carbonate.","The aqueous carbonate phase is then acidified with hydrochloric acid and extracted with methylene chloride and dried (Na2SO4).","The solvent is removed and the residue subjected to SiO2 column chromatography (hexane/ethyl acetate) affording 1-benzoyl-3,3,5-trimethylcyclohexan-2,6-dione 8.(W.","J. Michaely and G. W. Kraatz, EP-B-135191).","Dehydroangustione and xanthostemone derivatives are simply derived from dehydrogenation of angustione derivatives, for example, by treatment of 1-benzoyl-3,3,5-trimethylcyclohexan-2,6-dione 8 with palladium on charcoal in methanol, which thereby affords 1-benzoyl-3,5,5-trimethylcyclohex-3-en-2,6-dione 9 (Scheme 5).","Metal salts (enolates) of the above compounds can be prepared by the reaction of triketone derivatives with the corresponding metal hydroxides suspended in methanol or ethanol.","Trialkylammonium salts can be prepared by the reaction of triketone derivatives (e.g., 3) with trialkylamines in a chlorinated solvent such as dichloromethane.","Tetraalkylammonium salts can be prepared by adding a halogenated tetraalkylammonium salt to a metal salt in dichloromethane, which precipitates the metal halide removed by filtration.","The pure material is obtained by evaporation of the filtrate.","The present inventors have discovered that the β-diones of the invention can be obtained from natural sources and, in particular, from volatile oil-bearing organisms.","Accordingly, in another aspect, the present invention encompasses the use of a β-dione compound, particularly a β-diketone or a β-triketone compound, obtainable from a volatile oil-bearing organism, including an analogue or derivative thereof, in the preparation of a pesticidal composition for controlling harmful, annoying or undesired pests.","The present invention contemplates the use of any volatile oil-bearing organism that produces β-diones, preferably the β-diones according to any one of formulae (I)-(V), and especially β-diketones and/or β-triketones, for the preparation of the pesticidal compositions of the invention.","Preferred volatile oil-bearing organisms are volatile oil-bearing plants including, but not restricted to, plants from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.","Preferably, the volatile oil-bearing plant is selected from genera of the Myrtaceae family including, but not limited to, Angophora, Austromyrtus, Backhousia, Baeckea, Callistemon, Corymbia, Darwinia, Eucalyptus, Kunzea, Leptospermum, Melaleuca, Syzygium and Xanthostemon.","Thus, the compositions of the present invention may contain as active ingredients substantially purified β-diones or crude β-dione-containing extracts obtained from a volatile oil-bearing organism, preferably a volatile oil-bearing plant.","Volatile oils, also known in the art as essential oils, typically comprise a volatile mixture of esters, aldehydes, alcohols, ketones and terpenes, which can be prepared from botanical materials or plant cell biomass from cell culture.","Volatile oils can be prepared by subjecting botanical materials to a distillation process, for example.","A number of different procedures can be used for distillation.","For example, plant matter (e.g., foliage, stems, roots, seeds, bark etc) of a volatile oil-bearing plant is placed in a suitable still and steam distillation is used to break down the cells of the plant to release the oil.","The steam is then condensed and the oil phase is separated from the aqueous phase to obtain the volatile oil.","It will be appreciated that other methods of volatile oil extraction (e.g., solvent extraction) are known to those of skill in the art and it will be understood, in this regard, that the present invention is not limited to the use or practice of any one particular method of extracting volatile oils.","Suitably, the compositions comprise naturally-occurring compounds derived from a volatile oil-bearing organism.","Thus, in a preferred embodiment, the pesticidal composition of the invention comprises one or more β-dione active compounds, particularly β-diketone- and/or β-triketone-active compounds, that are derived from the volatile oil of a volatile oil-bearing organism.","In this embodiment, the composition may optionally contain a naturally-occurring carrier and/or other naturally-occurring additives.","Naturally-occurring additives contemplated by the present invention include natural antioxidants, which can be used advantageously to reduce the effect of oxidation of the compounds of the invention.","An example of a suitable naturally-occurring antioxidant is α-tocopherol.","Other additives, such as naturally-occurring stabilisers, are also contemplated, which may desirably be added to improve the stability and shelf life of the composition.","Examples of suitable natural stabilisers include gum arabic, guar gum, sodium caseinate, polyvinyl alcohol, locust bean gum, xanthan gum, kelgum, and mixtures thereof.","In an alternate embodiment, the naturally-occurring compounds derived from a volatile oil may be modified or derivatised to improve, for instance, their shelf-life, stability, activity and/or bioavailability.","The compounds of the present invention are useful for controlling harmful, annoying or undesired pests.","They may be used singularly or in combination with other pest-controlling compounds of the invention.","By “controlling” is meant preventing, combating, eradicating, destroying, repelling, or mitigating pests or increasing the mortality or inhibiting the growth and/or development of pests.","The term “pest” is used herein in its broadest sense and includes within its scope insects, arachnids (e.g.","acari, spiders), helminths (e.g., nematodes), molluscs, protozoa (e.g., Plasmodium sp.","Paramecium sp.","), viruses (e.g., herpesviruses) and the like.","Suitable applications for such control include, but are not limited to, combating and/or eradicating infestations by pests in animals (including humans) and/or plants (including trees) and/or stored products, which includes the administration to the animal or site of an effective quantity of a compound of the invention.","By “effective amount” is meant the administration or application of that amount of active compound, either in a single dose or as part of a series, that is effective for controlling a significant number of pests.","Thus, for example, a “pesticidally-effective” amount is the amount of active compound that is effective for increasing the mortality or decreasing the growth of a significant number of pests.","Alternatively, a “pest-repelling” effective amount is the amount of active compound that is noxious to, and/or induces behavioural changes in, a significant number of pests.","The effective amount will vary depending upon the taxonomic group of pest exposed to the active compound, the formulation of the composition, and other relevant factors.","It is expected that the amount will fall in a relatively broad range that can be determined through routine trials.","Accordingly, the compounds of the present invention can be used as pesticides, such as but not limited to insecticides, arachnicides, anti-helminthics, molluscicides antivirals, antiprotozoals and the like, or as pest repellents including repellents of insects, arachnids, helminths, molluscs, protozoa and viruses.","In especially preferred embodiments, the compounds of the present invention are used in the control of insects, arachnids, helminths or molluscs.","In practice, the compounds can be applied as formulations containing the various adjuvants and carriers known to or used in the industry for facilitating bioavailability, stability and dispersion.","The choice of formulation and mode of application for any given compound may affect its activity, and selection will be made accordingly.","In general, a pest-controlling compound of the invention can be compounded with appropriate inert carriers and additives in an appropriate ratio by means of dissolving, separating, suspending, mixing, impregnating, adsorbing or precipitating operation to formulate into oil formulations, emulsifiable concentrates, wettable powders, flowables, granules, powders, dusts, solutions, suspensions, emulsions, controlled-release forms such as microcapsules, aerosols or fumigants.","Typically, the compounds of the present invention can be mixed with a solid carrier, liquid carrier or gas carrier, optionally together with a surfactant and other adjuvants useful for such formulations.","The compounds of the invention can be used in an amount from about 0.00005% to about 90% by weight as contained in these formulations as their active component.","As used herein, the term “about” refers to a quantity, level, value or amount that varies by as much as 30%, preferably by as much as 20%, and more preferably by as much as 10% to a reference quantity, level, value or amount.","Where the compounds are in the form of β-dione-containing extracts, the formulations will usually comprise as their principal active ingredient from about 0.0001% to about 90%, preferably from about 0.0001% to about 50%, more preferably from about 0.0005% to about 10%, even more preferably from about 0.0005% to about 5%, even more preferably from about 0.0005% to about 1% and still even more preferably from about 0.001% to about 0.5% by weight of the extract.","Alternatively, where the compounds are in the form of substantially purified preparation of β-diones, the formulations will usually comprise as their principal active ingredient from about 0.00005% to about 90%, preferably from about 0.0001% to about 50%, more preferably from about 0.0005% to about 10%, even more preferably from about 0.001% to about 5% and still even more preferably from about 0.001% to about 1% by weight of the substantially purified β-dione.","By “substantially purified” is meant a compound which has been separated from components that naturally accompany it.","Typically, a compound is substantially pure when at least 60%, more preferably at least 75%, more preferably at least 90%, and most preferably at least 99% of the total material (by volume, by wet or dry weight, or by mole percent or mole fraction) in a sample is the compound of interest.","Purity can be measured by any appropriate method, e.g., by chromatography or HPLC analysis.","Examples of solid carriers useful in preparing the formulations are clays including kaolin clay, diatomite, water-containing synthetic silicon oxide, bentonite, Fubasami clay, and acid clay; talcs; ceramics; inorganic minerals such as Celite, quartz, sulfur, active carbon, calcium carbonate and hydrated silica; and chemical fertilisers such as ammonium sulfate, ammonium phosphate, ammonium nitrate, urea and ammonium chloride, these solid carriers being finely divided or granular.","Examples of useful liquid carriers are water, alcohols such as methanol and ethanol, ketones such as acetone and methyl ethyl ketone, aromatic hydrocarbons such as benzene, toluene, xylene, ethylbenzene and methylnaphthalene, aliphatic hydrocarbons such as hexane, cyclohexane, kerosene and light oil, esters such as ethyl acetate and butyl acetate, nitriles such as acetonitrile and isobutyronitrile, ethers such as diisopropyl and dioxane, acid amides such as N,N-dimethylformamide and N,N-dimethylacetamide, halogenated hydrocarbons such as dichloromethane, trichloroethane and carbon tetrachloride, dimethyl sulfoxide, and fish oils, mineral oils, plant derived oils such as canola oil, cotton-seed oil, soybean oil and sesame oil as well as essential oils such as lavender oil, eucalyptus oil, tea tree oil, citrus oil etc.","Solid or liquid carriers can be used alone or in combination.","Examples of gas carriers, i.e., those of propellants, are butane gas, LPG (liquefied petroleum gas), dimethyl ether, fluorocarbons and carbon dioxide gas.","Examples of surfactants are alkylsulfuric acid esters, alkylsulfonic acid salts, alkylarylsulfonic acid salts, alkyl aryl ethers and polyoxyethylene adducts thereof, polyethylene glycol ethers, polyhydric alcohol esters, sugar alcohol derivatives, sorbitane monolaurate, alkylallyl sorbitane monolaurate, alkylbenzene sulfonate, alkylnaphthalene sulfonate, lignin sulfonate, and sulfuric acid ester salts of higher alcohols.","These surfactants may be used alone or in combination.","Examples of adjuvants for the formulations, such as binders and dispersants, are casein, gelatin, polysaccharides such as starch, gum arabic, cellulose derivatives and alginic acid, lignin derivatives, bentonite, sugars and water-soluble synthetic high-molecular-weight substances such as polyvinyl alcohol, polyvinyl pyrrolidone and polyacrylic acids.","Examples of stabilisers are PAP (acid isopropyl phosphate), BHT (2,6-di-tert-butyl-4-methylphenol), BHA (mixture of 2-tert-butyl-4-methoxyphenol and 3-tert-butyl-4-methoxyphenol), synergists such as piperonyl butoxide, vegetable oils, mineral oils, fish oils, surfactants and fatty acids or esters thereof.","Emulsifying agents that may be used are suitably one or more of those selected from non-ionic or anionic emulsifying agents.","Examples of non-ionic emulsifying agents include, but are not restricted to, polyoxyethylenealkylphenylether, polyoxyethylenealkylether, polyethyleneglycol fatty ester, sorbitan fatty ester, polyoxyethylene sorbitan fatty ester, polyoxyethylenesorbitol fatty ester, polyoxyethylenepolyoxypropylenealkylether.","Examples of anionic emulsifying agents include alkyl sulphates, polyoxyethylenealkylether sulphates, sulfosuccinates, taurine derivatives, sarcosine derivatives, phosphoric esters, alkylbenzenesulfonates and the like.","A mixture consisting of polyoxyethylenestyrylphenylether and calcium allylbenzenesulfonate is preferred.","These emulsifying agents may be used in an amount of 5 to 20 weight parts per 100 weight parts of the compositions of the present invention.","Formulation thus obtained can be used solus or diluted, for example, with water or other diluent.","The formulations can be used also as admixtures with other pesticides such as insecticides, arachnids, anti-helminthics, molluscicides, herbicides, plant growth regulators, synergists, soil improvers, baits and the like, or can be used simultaneously with such agents without mixing.","For example, the pest-controlling compounds of the invention can be combined with other naturally derived bioactive compounds or extracts such as neem or its components, derris, pyrethrum; microbial extracts such as avermectins or streptomycins; with synthetic insecticides, acaricides, molluscicides, anti-helminthics; anti-protozoals, antivirals or with microorganisms having insecticidal, acaricidal, molluscicidal, anti-helminthic anti-protozoal or antiviral activity e.g., bacteria such as Bacillus thuringiensis, Bacillus popillae, entomogenous fungi such as Metarhizium spp., Verticillium lecanii, nematodes such as Steinernema spp and Heterorhabditis.","Alternatively, or in addition, the pest-controlling compounds of the invention can be combined with synergists such as piperonyl butoxide, and with ultraviolet screening compounds of natural or synthetic origin.","When used as an agricultural pesticide, the compound of the invention is preferably applied usually in an amount of 0.01 to 500 g/100 m2.When an emulsifiable concentrate, wettable powder or flowables are used as diluted with water, the compound is applied usually at a concentration of 0.1 to 1000 ppm, preferably 1 to 500 ppm.","The granular or dust can be applied without dilution.","The amount or concentration of application, although exemplified above, can be suitably increased or reduced according to the type of preparation, time, place, method of application, kind of pest and extent of harm or annoyance suffered.","The invention also contemplates the use of the above described β-dione compounds in pest repellent, particularly insect repellent, compositions.","Repellent compositions contemplated by the present invention include those that are noxious to, and/or induce behavioural changes in, a pest.","The latter compositions suitably comprise an activity including, but not restricted to, an antifeedant activity, an oviposition deterrent activity and an insect growth regulatory activity.","Insect repellent compositions in various dosage forms can be prepared by blending the above-described β-dione compounds as active ingredients with a base of cosmetics or pharmaceuticals, which are usually applied to human bodies or animals.","They can be formulated in, for example, lotions, aerosols, milky lotions, creams or the like.","These compounds can be further incorporated with other insect repellents, antioxidants, UV-absorbers, humectants or other additives.","The above compounds or the above-prepared compositions of the present invention can be applied directly to human bodies or animals.","Besides, substrates, such as sheets, films, nets, timber or the like, which have preliminarily been treated with the above compounds or compositions by means of application, impregnation or blending, can also be used.","The quantity of the above compounds to be formulated in the noxious-insect repellents depends upon the dosage form, usage or other conditions.","Suitable dosages may be selected from about 0.1% to about 90% by weight.","Thus, in another aspect of the present invention there is provided a method for controlling harmful, annoying or undesired pests, said method comprising exposing said pests to a pest-controlling effective amount of a composition comprising a β-dione compound as broadly described above.","Preferred embodiments of this type include exposing said pests to a pesticidally effective amount or a pest-repelling effective amount of said composition.","The terms “comprise”, “comprises” and “comprising” and the like refer, unless the context requires otherwise, to the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements.","The compositions and methods of the present invention may be applied to pests including, but not restricted to, insects, arachnids, helminths, molluscs, protozoa and viruses.","For example, suitable insects that fall within the scope of the present invention include those: (a) from the order of the lepidopterans (Lepidoptera), for example, Adoxophyes orana, Agrotis upsilon, Agrotis segetum, Alabama argillacea, Anticarsia gemmatalis, Argyresthia conjugella, Autographa gamma, Cacoecia murinana, Capua reticulana, Choristoneuraffumiferana, Chilo partellus, Choristoneura occidentalis, Cirphis unipuncta, Cnaphalocrocis medinalis, Crocidolomia binotalis, Cydia pomonella, Dendrolimus pini, Diaphania nitidalis, Diatraea grandiosella, Earias insulana, Elasmopalpus lignosellus, Eupoecilia ambiguella, Feltia subterranea, Grapholitha funebrana, Grapholitha molesta, Heliothis armigera, Heliothis virescens, Heliothis zea, Hellula undalis, Hibernia defoliaria, Hyphantria cunea, Hyponzomeuta malinellus, Keiferia lycopersicella, Lambdina fiscellaria, Laphygma exigua, Leucoptera scitella, Lithocolletis blancardella, Lobesia botrana, Loxostege sticticalis, Lymnantria dispar, Lymantria monacha, Lyonetia clerkella, Manduca sexta, Malacosoma neustria, Mamestra brassicae, Mocis repanda, Operophthera brumata, Orgyia pseudotsugata, Ostrinia nubilalis, Pandemis heparana, Panolisflammea, Pectinophora gossypiella, Phthorimaea operculella, Phyllocnistis citrella, Pieris brassicae, Plathypena scabra, Platynota stultana, Plutella xylostella, Prays citri, Prays oleae, Prodenia sunia, Prodenia ornithogalli, Pseudoplusia includens, Rhyacionia fiustrana, Scrobipalpula absoluta, Sesamiainferens, Sparganothis pilleriana, Spodoptera fugiperda, Spodoptera littoralis, Spodoptera litura, Syllepta derogata, Synanthedon myopaeformis, Thaumatopoea pityocampa, Tortrix viridana, Trichoplusia ni, Tryporyza izcertulas, Zeiraphera canadensis, (b) furthermore Galleria mellonella and Sitotroga cerealella, Ephestia cautella, Tineola bisselliella; (c) from the order of the beetles (Coleoptera), for example, Anthonomus grandis, Anthonomus pomorum, Apion vorax, Atomaria linearis, Blastophagus piniperda, Cassida nebulosa, Cerotoma trifurcata, Ceuthorhynchus assimilis, Ceuthorhynchus napi, Chaetocnema tibialis, Conoderus vespertinus, Crioceris asparagi, Dendroctonus refipennis, Diabrotica longicornis, Diabrotica 12-punctata, Diabrotica virgifera, Epilachna varivestis, Epitrix hirtipennis, Eutinobothrus brasiliensis, Hylobius abietis, Hypera brunneipennis, Hypera postica, Ips typographus, Lema bilineata, Lema melanopus, Leptinotarsa decemlineata, Limonius californicus, Lissorhoptrus oryzophilus, Melanotus communis, Meligethes aeneus, Melolontha hippocastani, Melolontha melolontha, Oulema oryzae, Ortiorrhynchus sulcatus, Otiorrhynchus ovatus, Phaedon cochleariae, Phyllopertha horticola, Phyllophaga sp., Phyllotreta chrysocephala, Phyllotreta nemorum, Phyllotreta striolata, Popillia japonica, Psylliodes napi, Scolytus intricatus, Sitona lineatus, (d) furthermore Bruchus rufimanus, Bruchus pisorum, Bruchus lentis, Sitophilus granaria, Lasioderna serricorne, Oryzaephilus surinamensis, Rhyzopertha dominica, Sitophilus oryzae, Tribolium castaneum, Trogodernia granarium, Zabrotes subfasciatus; (e) from the order of the dipterans (Diptera), for example, Anastrepha ludens, Ceratitis capitata, Contarinia sorghicola, Dacus cucurbitae, Dacus oleae, Dasineura brassicae, Delia coarctata, Delia radicum, Hydrellia griseola, Hylemyia platura, Liriomyza sativae, Liriomyza trifolii, Mayetiola destructor, Orseolia oryzae, Oscinellafrit, Pegomya hyoscyami, Phorbia antiqua, Phorbia brassicae, Phorbia coarctata, Rhagoletis cerasi, Rhagoletis pomonella, (f) furthermore Aedes aegypti, Aedes vexans, Anopheles maculipennis, Chrysomya bezziana, Chrysomya hominivorax, Chrysomya macellaria, Cordylobia anthropophaga, Culex pipielis, Fannia canicularis, Gasterophilus intestinalis, Glossina morsitans, Haematobia irritans, Haplodiplosis equestris, Hypoderma lineata, Lucilia caprina, Lucilia cuprina, Lucilia sericata, Musca domestica, Muscina stabulans, Oestrus ovis, Tabanus bovinus, Simulium damnosum; (g) from the order of the thrips (Thysanoptera), for example, Frankliniella fusca, Frankliniella occidentalis, Frankliniella tritici, Haplothrips tritici, Heliothrips haemorrhoidalis, Scirtothrips citri, Thrips oryzae, Thripspalmi, Thrips tabaci; (h) from the order of the hymenopterans (Hymenoptera), for example, Athalia rosae, Atta cephalotes, Atta sexdens, Atta texana, Hoplocampa minuta, Hoplocampa testudinea, Iridomyrmes humilis, Iridomyrmex purpureus, Monomorium pharaonis, Solenopsis geminata, Solenopsis invicta, Solenopsis richteri, Technomyrmex albipes; (i) from the order of the heteropterans (Heteroptera), for example, Acrosternum hilare, Blissus leucopterus, Cyrtopeltis notatus, Dysdercus cingulatus, Dysdercus intermedius, Eurygaster integriceps, Euschistus impictiventris, Leptoglossus phyllopus, Lygus hesperus, Lygus lineolaris, Lygus pratensis, Nezara viridula, Piesma quadrata, Solubea insularis, Thyanta perditor; (j) from the order of the homopterans (Homoptera), for example, Acyrthosiphon onobrychis, Acyrthosiphon pisum, Adelges laricis, Aonidiella aurantii, Aphidula nasturtii, Aphis fabae, Aphis gossypii, Aphis pomi, Aulacorthum solani, Bemisia tabaci, Brachycaudus cardui, Brevicoryne brassicae, Dalbulus maidis, Dreyfusia nordmannianae, Dreyffusia piceae, Dysaphis radicola, Empoasca fabae, Eriosonma lanigerum, Laodelphax striatella, Macrosiphum avenae, Macrosiphum euphorbiae, Macrosiphon rosae, Megoura viciae, Metopolophium dirhodum, Myzus persicae, Myzus cerasi, Nephotettix cincticeps, Nilaparvata lugens, Perkinsiella saccharicida, Phorodon humuli, Psylla mali, Psylla piri, Psylla pyricola, Rhopalosiphum maidis, Schizaphis graminum, Sitobion avenae, Sogatella furcifera, Toxoptera citricida, Trialeurodes abutilonea, Trialeurodes vaporariorum, Viteus vitifolii; k) from the order of the termites (Isoptera), for example, Calotermes flavicollis, Coptotermes spp, Leucotermes flavipes, Macrotermes subhyalinus, Nasutitermes spp such as Nasutitermes walkeri, Odontotermes formosanus, Reticulitermes lucifugus, Termes natalensis; (l) from the order of the orthopterans (Orthoptera), for example, Gryllotalpa gryllotalpa, Locusta migratoria, Melanoplus bivittatus, Melanoplus femur-rubrum, Melanoplus mexicanus, Melanoplus sanguinipes, Melanoplus spretus, Nomadacris septemfasciata, Schistocerca americana, Schistocerca peregrina, Stauronotus maroccanus, Schistocerca gregaria, (m) furthermore Acheta domestica, Blatta orientalis, Blattella germanica, Periplaneta americana; (n) from the order of the phthirapterans (Phthiraptera), for example, Mallophaga, such as Damalina spp., and Anoplura such as Linognathus and Haematopinus spp.","; (O) from the order of the hemipterans (Hemiptera), for example, Aphis, Bemisia, Phorodon, Aeneolamia, Empoasca, Parkinsiella, Pyrilla, Aonidiella, Coccus, Pseudococcus, Helopeltis, Lygus, Dysdercus, Oxycarenus, Nezara, Aleyrodes, Triatoma, Psylla, Myzus, Megoura, Phylloxera, Adelges, Nilaparvata, Nephotettix or Cimwx spp.","; (p) from the order of the siphonapterans (Siphonaptera), for example, Ctenocephalides or Pulex spp.","; (q) from the order of the thysanurans (Thysanura), for example, Lepisma spp.","; (r) from the order of the dermapterans Dermaptera), for example, Forficula spp.","; and (s) from the order of the psocopterans (Psocoptera), for example, Peripsocus spp.","Arachnids contemplated by the present invention include, but are not limited to, spiders and scorpions and especially mites such as phytophagous mites (Acari), such as Aculops lycopersicae, Aculops pelekassi, Aculus schlechtendali, Brevipalpus phoenicis, Bryobia praetiosa, Eotetranychus carpini, Eutetranychus banksii, Eriophyes sheldoni, Oligoinychus pratensis, Panonychus ulmi, Panonychus citri, Phyllocoptruta oleivora, Polyphagotarsonenmus latus, Tarsonemuspallidus, Tetranychus cinnabarinus, Tetranychus kanzawai, Tetranchus pacificus, Tetranychus urticae, ticks, such as Amblyomma americanum, Amblyomma variegatum, Argas persicus, Boophilus annulatus, Boophilus decoloratus, Boophilus microplus, Dermacentor silvarum, Hyalomma truncatum, Ixodes ricinus, Ixodes rubicundus, Ornithodorus moubata, Otobius megnini, Rhipicephalus appendiculatus and Rhipicephalus evertsi, and animal-parasitic mites, such as Dermanyssus gallinae, Psoroptes ovis and Sarcoptes scabiei.","Helminths falling within the scope of the present invention may be selected from cestodes such as fish tapeworm, pork tapeworm, beef tapeworm, and dwarf tapeworm; trematodes such as from the genera Metagonimus and Heterophyes; and nematodes such as but not limited to filariid, ascarid, strongyle and trichostrongyle nematodes of the genera Acanthocheilonema, Aelurostrongylus, Ancylostoma, Angiostrongylus, Ascaris, Brugia, Bunostomum, Dictyocaulus, Dioctophyme, Dipetalonema, Dirofilaria, Dracunculus, Filaroides, Lagochilascaris, Loa, Mansonella, Muellerius, Necator, Onchocerca, Parafilaria, Parascaris, Protostrongylus, Setaria, Stephanofilaria, Strongyloides, Strongylus, Thelazia, Toxascaris, Toxocara, Trichinella, Uncinaria and Wuchereria.","Suitable molluscs include those of the Gastropoda class examples of which include snails, slugs, conchs, and whelks.","Protozoa may be selected for example from Plasmodia, Toxoplasma, Leishmania, Trypanosoma, Giardia, Entamoeba, Acanthamoeba, Nagleria, Hartmanella, Balantidium, Babesia, Cryptosporidium, Isospora, Microsporidium, Trichomonas or Pneumocystis species.","Viruses may be selected from RNA viruses or DNA viruses, which include but are not limited to Human Immunodeficiency Virus (HIV), Poliovirus, Influenza virus, Rous Sarcoma virus, Flaviviruses such as Japanese encephalitis, Influenza virus, Respiratory Syncytial Virus, Hepatitis virus, Parvovirus, Rotavirus, Coronavirus, Adenovirus and Herpesviruses such as Papillomavirus and Epstein-Barr virus.","The present invention also extends to methods for producing resistance in plants to pests including, but not limited to, insects, arachnids, helminths, molluscs, protozoa and viruses by crossing a plant expressing a β-dione compound according to the invention with pest susceptible lines.","Crossing a β-dione-producing plant into a pest susceptible background would produce a resistant plant with a high level of pest resistance.","Plants that could be made pest resistant include, but are not limited to, dicotyledonous plants, especially trees and more especially members of the Myrtaceae family.","For example E. cloeziana commonly known as Gympie Messmate is one of the many Eucalyptus species grown for hard wood production.","However the oil present in this chemotype does not contain β-diones and hence an intra species cross with the unique North Queensland tasmanone chemotype would introduce this phenotypic trait.","Such a process would be readily applicable to other Eucalyptus species of commercial interest.","Interspecific crossing within the Myrtaceae family is well established to those skilled in the art and inclusion of β-dione as an additional trait into formal breeding programs is acknowledged.","Suitable β-dione-producing plants may be selected from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.","Preferably, the volatile oil-bearing plant is selected from genera of the Myrtaceae family including, but not limited to, Angophora, Austromyrtus, Backhousia, Baeckea, Callistemon, Corymbia, Darwinia, Eucalyptus, Kunzea, Leptospermum, Melaleuca, Syzygium and Xanthostemon.","Preferred 16-dione-producing plants are Leptospermum mornsonii, Eucalyptus bensonii, Eucalyptus megacornuta, Eucalyptus pilularis, Eucalyptus cornuta, Eucalyptus baxteri, Eucalyptus macrorhyncha, Eucalyptus cloeziana, Melaleuca cajuputi, Eucalyptus jensenii, Backhousia angustifolia and Leptospennum scoparium.","A particularly preferred β-dione-producing plant is Eucalyptus cloeziana.","As used herein, the term “plant” includes reference to whole plants, plant organs (e.g., leaves, stems, roots, etc.","), seeds and plant cells and progeny of same.","Plant cell, as used herein includes, without limitation, seeds suspension cultures, embryos, meristematic regions, callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen, and microspores.","The class of plants which can be used in the methods of the invention is generally as broad as the class of higher plants amenable to transformation techniques, including both monocotyledonous and dicotyledonous plants.","Thus, the present invention contemplates conventional plant breeding methods to transfer the genetic material associated with β-dione production via crossing and backcrossing.","Such methods will comprise the steps of: (1) sexually crossing the β-dione-producing plant with a plant from a pest susceptible taxon; (2) recovering reproductive material from the progeny of the cross; and (3) growing β-dione-producing/pest-resistant plants from the reproductive material.","Where desirable or necessary, the agronomic characteristics of the susceptible taxon can be substantially preserved by expanding this method to include the further steps of repetitively: (1) backcrossing the pest-resistant progeny with pest-susceptible plants from the susceptible taxon; and (2) selecting for expression of a β-dione (or an associated marker gene) among the progeny of the backcross, until the desired percentage of the characteristics of the susceptible taxon are present in the progeny along with the gene or genes imparting β-dione activity.","By the term “taxon” herein is meant a unit of botanical classification.","It thus includes, genus, species, cultivars, varieties, variants and other minor taxonomic groups which lack a consistent nomenclature.","In order that the invention may be readily understood and put into practical effect, particular preferred embodiments will now be described by way of the following non-limiting examples.","EXAMPLES Example 1 β-Triketone-Containing Oils Obtained from Australian Myrtaceae Species Australia has an extensive number of volatile oils from species of the Myrtaceae that are rich in a diversity of structurally related constituents known as β-triketones.","These oils often show not only a high yield of oil, but also a high degree of biosynthetic selectivity that produces β-triketones in a high proportion of the oil composition.","The major constituents of the published Myrtaceae essential oils (Hellyer, 1968; Boland and Brophy, 1990, 1993; Brophy, et al., 1995; Bignall et al., 1997; Southwell and Brophy 2000) are listed in Table 1 and their structures are included in FIG.","1.TABLE 1 β-TRIKETONE PROFILES OF AUSTRALIAN ESSENTIAL OILS Species Yield β-Triketone (%) Distribn Backhousia angustifolia 2.5 Angustione (85) QLD Backhousia angustifolia 2.5 Dehyroangustione (80) QLD Eucalyptus cloeziana 3.0 Tasmanone (95) QLD Eucalyptus suberea 1.4 Tasmanone (94) WA Eucalyptus lateritica 0.9 Tasmanone (37), WA lateriticone (14) Eucalyptus camfieldii Tasmanone (40) NSW Leptospermum scoparium 0.4 Leptospermone (19), NSW, NZ Flavesone (8), Isoleptospermone (5) Eucalyptus grandis 0.6 Leptospermone (20), NSW Flavesone (13), Isoleptospermone (3) Eucalyptus agglomerata Agglomerone (40) NSW Eucalyptus mckieana Agglomerone (60) NSW Eucalyptus bensonii 2.5 Agglomerone (72) Eucalyptus insularis 1 Agglomerone (19) WA Eucalyptus jensenii 0.3 Jensenone (70) NT Eucalyptus papuana 0.7 Papuanone (40) Nth Aus Leptospermum morrisonii 1.8 Grandiflorone (58) NSW Melaleuca cajeputi Platyphyllol Nth Aus Xanthostemon Xanthostemone oppositifolius The β-triketones obtained from selected Myrtaceous volatile oils were shown to have significant insecticidal and/or acaricidal activity.","Example 2 Insecticidal Activity Initial insecticidal screening against two important arthropod species, two-spotted mite (Tetranychus urticae) and diamond back moth (Plutella xylostella) lavae highlighted three oils on the basis of efficacy, oil yield, oil profile and ease of recollection (Table 2).","Where feasible, the LD50 and LD95 values were determined.","TABLE 2 PERCENTAGE MORTALITY OF THREE EFFICACIOUS OILS Diamond Two Spotted Mite Back Moth % Mortality % Mortality Species (0.5%/1.0%) (0.5%/1.0%) Backhousia angustifolia 98/98 100/100 Backhousia angustifolia 100/100 100/100 Eucalyptus cloeziana 100/100 70/100 Insecticidal tests using the oil from fresh plant recollections and steam distillations varied occasionally and, in this regard, it is believed that improving storage conditions including temperature, light and exposure to air and inclusion of a dessicant can enhance the stability of the active fraction of such oils.","E. cloeziana oil continued to show high potency against both insect tests.","This oil exhibited an LD95 between 0.04-0.20% (depending on formulation and treatment) against two-spotted mite.","An LD95 of 0.10% was observed against 1st instar lavae of diamond back moth and this rose to 0.78% when tested against 3rd instar lavae.","In additional preliminary investigations with greenhouse thrips (Heliothrips haemorrhoidalis), a 0.1% concentration of E. cloeziana oil induced 100% mortality.","Additional work was carried out on the E. cloeziana oil to explore the contribution the various components make to the overall efficacy of this oil.","Fractions 1, 3, 4, and 5 outlined in Example 3 were screened against two-spotted mite.","Fractions 1 and 5 showed no significant insecticidal effect.","Fractions 3 and 4, comprising 98% and 99% tasmanone were active and showed little difference to the activity of the whole oil.","This suggests that not only is tasmanone the principle component in the oil, but it is also the principle bioactive constituent.","It is also reasonable to assume that as the activity of E. cloeziana oil has been demonstrated against a number of different arthropod species, namely a mite (T. urticae), a caterpillar (P. xylostella) and a thrips (H. haemorrhoidalis), its insecticidal activity is broad in nature.","Example 3 Chemistry Chemical analysis (GC-MS) of the steam-distilled oils from the collected plants in this work are summarised in Table 3.TABLE 3 β-TRIKETONE PROFILES OF SELECTED OILS Plant Source Principle Component* (%) Backhousia angustifolia-I Dehydroangustione 85% Backhousia angustifolia-II Dehydroangustinone (80%) Backhousia angustifolia-III Angustinone (65%) Backhousia angustifolia-IV Angustinone (28%) Eucalyptus cloeziana Tasmanone (84-96%) Melaleuca cajuputi subsp platyphylla Platyphyllol (64-71%) *As determined by gas chromatography The most promising oils were derived from B. angustifolia, M. cajuputi subsp platyphylla and E. cloeziana and these were then subjected to additional chemical fractionation.","The lower insecticidal activity observed in the recollected B. angustifolia (I) was in part attributed to the lower levels of β-triketone.","The level of β-triketone was elevated by the removal of the more volatile monoterpenes using vacuum distillation.","One of the most efficacious oils was from E. cloeziana, a tasmanone (84-96%) rich oil with additional terpenes and β-triketones (FIG.","2, Table 4), which displayed consistent activity at every stage of processing and formulation.","This oil was, therefore, fractionated using column chromatography on silica gel with a hexane-diethyl ether gradient and a final methanol elution.","The profiles of the fractions are summarised in Table 5.TABLE 4 CHEMICAL PROFILE OF E. cloeziana OIL Peak No Compound Composition (%) 1 a-pinene 0.5-1.9 2 B-pinene 1.5-5.7 3 Limonene 0.1-0.6 4 a-terpineol 0.7-2.0 5 Globulol 0.01-0.5 6 Agglomerone 0.01-0.6 7 Tasmanone 84-96 8 Lateriticone 0.2-0.7 9 Isolateriticone 0.3-1.2 TABLE 5 FRACTIONS CUT FROM E. cloeziana OIL Fraction No Solvent System Composition Amount F1 Hexane Hydrocarbons 73 mg F2 Hex:Et2O (9:1) 80% Tasmanone 4 mg F3 Hex:Et2O (1:1) 98% Tasmanone 3.66 g F4 Hex:Et2O (9:1) +99% Tasmanone 694 mg F5 MeOH Terpene alcohols 64 mg 1H NMR data (FIG.","3) were recorded on F4 and confirmed the structure of tasmanone.","Moreover the compound exists in solution (CDCl3) in tautomeric forms (FIG.","4) in the ratio 2:0:1 (A:B:C).","Another efficacious oils was from M. cajuputi subsp platyphylla, a platyphyllol (64-71%) rich oil (Table 6), which also displayed consistent activity at every stage of processing and formulation.","TABLE 6 TYPICAL CHEMICAL PROFILE OF M. cajuputi subsp platyphylla OIL Peak No Compound Composition (%) 1 a-pinene Tr-0.8 2 1,8-cineole Tr-0.7 3 B-caryophyllene 0.6-3.2 4 Humulene 0.6-1.3 5 Spathulenol 4.0-9.0 6 Caryophyllene oxide Tr-3.6 6 Platyphyllol 64-71 7 MW 234 - unknown 4.3 *As determined by gas chromatography Example 4 Phytotoxicity Initial investigations using leaves and leaf discs of several plant species including French bean (Phaseolus vulgaris) lemon (Citrus limon) Orange (Citrus sinensis) and Cabbage (Brassica oleracea) indicated that phytotoxicity did not occur for most oils below 0.5%.","More detailed investigations using soft intact leaves of young greenhouse-grown French bean subsequently showed E. cloeziana oil applied as a spray caused some phytotoxicity at concentrations of 0.5% and above.","Example 5 Toxicity of E. Cloeziana Oil Bacterial Reverse Mutation Assay This study investigated the potential of E. cloeziana oil to induce reverse mutations at the histidine locus in the genome of one strain of Salmonella typhimurium TA100 in the presence and absence of a metabolic activation system (mammalian microsomal enzymes, S9 mix).","The test sample was dissolved in dimethyl sulfoxide (DMSO).","In this assay, an E. cloeziana oil test sample did not induce an appropriate-fold increase (a 2-fold increase for TA100) in the mean revertants per plate in the tester strain TA100 over the mean revertants per plate of the appropriate vehicle control.","Accordingly, the test sample was considered to be non-mutagenic under the conditions of the assay.","Acute Oral Sighting The acute oral toxicity of E. cloeziana oil was investigated in ten (10) Sprague Dawley Specific Pathogen Free female rats (groups of 2) at doses of 500, 250, 125 and 50 mg/kg.","The experimental procedure was based on OECD guidelines for the testing of chemicals, No.","420.Clinical signs of toxicity occurred between one (1) and twenty-four (24) hours after dosing.","Both animals in the 500 mg/kg group exhibited subdued behaviour, partial eye closure, slow breathing, reduced motor activity, ataxia followed by death within 24 hours.","The animals in the 250 mg/kg group exhibited subdued behaviour, partial eye closure, slow breathing, social isolation, and reduced motor activity, and had returned to normal by 24 hours after dosing.","The animals in the 125 mg/kg group exhibited subdued behaviour, partial eye closure, slow breathing, piloerection and reduced motor activity, and had returned to normal by 24 hours after dosing.","The animals in the 50 mg/kg group did not show any signs of toxicity during the seven day experimental period.","There were no other clinical abnormalities in any animal throughout the seven (7) day observation period.","The stomach of one animal in the 250 mg/kg group (5F) had a single ulcer.","There were no other gross abnormalities in the major organs of any animal at autopsy.","Based on the results obtained from this study 50, 100 and 200 mg/kg can be selected for a main study as the maximum non-toxic dose, intermediate dose and high dose, respectively.","Full Study The acute oral toxicity of E. cloeziana oil was investigated in thirty (30) Sprague Dawley Specific Pathogen Fee rats (15 males and 15 females) at doses of 50, 100 and 200 mg/kg.","These doses were chosen following a dose range finding in the above study.","The experimental procedure was based on OECD guidelines for the testing of chemicals, No.","420.The acute NOAEL of E. cloeziana oil was determined to be 50 mg/kg, and the MTD was 200 mg/kg under the conditions of this study.","Acute Dermal The acute dermal toxicity of E. cloeziana oil was investigated in ten (10) Sprague Dawley Specific Pathogen Free rats (5 males and 5 females) at a dose of 2000 mg/kg.","A preliminary study (SIGHTING) indicated no signs of toxicity at this dose.","The experimental procedure was based on OECD guidelines for the testing of chemicals, No.","420.No clinical abnormalities, skin irritations or body weight losses were observed in any animal throughout the fourteen (14) day observation period.","No deaths occurred.","No abnormalities were seen in the major organs at necropsy.","The rat acute dermal LD50 of E. cloeziana oil was determined to be greater than 2000 mg/kg under the conditions of this study.","Skin Irritancy/Corrosion The potential of a test sample of E. cloeziana oil to provoke skin irritation/corrosion reactions was investigated using a primary dermal irritation/corrosion test in three (3) New Zealand White albino rabbits (OECD Guidelines for the Testing of Chemicals, No.","404).","The results obtained from this study indicated that E. cloeziana oil is a non-irritant according to the National Occupational Health and Safety Commission (NOHSC) “Approved Criteria for Classifying Hazardous Substances [NOHSC:1008(1999)]”.","Example 6 Isolation and Purification of β-Triketones from Australian Myrtaceae Species Plant collections were commissioned at the Mt.","Annan Botanic Garden, Sydney, Australia, and the Darwin Botanic Garden, Northern Territory, Australia to provide potential sources of a range of β-triketone isolation.","The plants investigated for β-triketone exploration are listed in Table 6.TABLE 6 PLANTS SOURCED FOR β-TRIKETONE EXPLORATION β-triketone # Source Plant Plant Location Accession # anticipated 1 Leptospermum morrisonii Mt.","Annan, NSW 873247 grandiflorone 2 Eucalyptus bensonii Mt.","Annan, NSW 881045 agglomerone 3 Eucalyptus megacornuta Mt.","Annan, NSW 831078 jensenone+ 4 Eucalyptus pilularis Mt.","Annan, NSW 873166 torquatone 5 Eucalyptus cornuta Mt.","Annan, NSW 852618 jensenone+ 6 Eucalyptus baxteri Mt.","Annan, NSW 860999 agglomerone/ tasmanone 7 Eucalyptus macrorhyncha Mt.","Annan, NSW 860896 conglomerone 8 Eucalyptus cloeziana Lappa, QLD PF 2513 tasmanone 9 Melaleuca cajuputi ssp Bensbach, WP, PNG KW16-19 platyphyllol platyphyla 10 Eucalyptus jensenii NT Botanic Gardens, RK114- jensenone Darwin, NT LUS 11 Backhousia angustifolia Wilgavale, Texas, PF1712 dehydroangustione QLD 12 Backhousia angustifolia Didcott Creek, QLD PF 1708 angustione 13 Leptospermum scoparium NA commercial oil NA flavesone, sample isoleptospermone, leptospermone 14 Eucalyptus conjuncta Mt Annan 854097 conglomerone +Oil contains other β-triketone constituents All plants, apart from 9 and 13, were steam distilled to yield various quantities of essential oil.","All oils were analysed by Gas Chromatography Mass Spectrometry (GCMS) to determine the presence and abundance of β-triketones.","Based on this information, particular oils were targeted for isolation and purification of β-triketones using wet chemistry and preparative HPLC techniques.","The β-triketones listed in Table 7 were isolated in quantities adequate for insecticidal screening.","A minimum of 50 mg of each compound was required.","Isoleptospermone and leptospermone were difficult to separate due to their structural similarity as were angustione and dehydroangustione.","Consequently, mixtures of these compounds, where one isomer was significantly more abundant, were provided for insecticidal screening as this will still allow for differentiation in observed activity.","TABLE 7 β-TRIKETONES ISOLATED FOR INSECTICIDAL SCREENING Amount sent Sample Purity % by for screening β-triketones Plant source GCMS (mg) grandiflorone Leptospermum morrisonii 100.00 99.5 jensenone Eucalyptus jensenii 100.00 67.9 dehydroangustione Backhousia angustifolia 95.02 165.02 PF1712 angustione Backhousia angustifolia 66.30 (33.70% 107.1 PF1708 dehydroangustione) agglomerone Eucalyptus bensonii 99.28 109.2 flavesone Leptospermum scoparium 99.32 101.8 isoleptospermone Leptospermum scoparium 33.10 (66.90% 113.4 leptospermone) leptospermone Leptospermum scoparium 95.33 77.0 tasmanone Eucalyptus cloeziana 99.92 110.1 platyphyllol Melaleuca cajuputi subsp.","99.62 255.6 platyphylla The chemical structures and identities of the β-triketones isolated were confirmed by GCMS and Nuclear Magnetic Resonance (NMR) analysis.","Example 7 Efficacy of E. cloeziana Oil on Target Organisms Target Organisms Two spotted mite (TSM) Tetranychus urticae Koch [Acarina: Tetranychidae] were collected from a mass culture maintained at the University of Western Sydney's Hawkesbury Campus in Richmond, NSW, Australia They were reared on potted French beans (Phaseolus vulgaris L [Fabales: Fabaceae] in a glasshouse maintained at 25±5° C., 65±5% RH and 14 h D:L photoperiod.","Only young females were selected for bioassay.","Adult parthenogenetic female greenhouse thrips (GHT), Heliothrips haemorrhoidalis Bouché (Thysanoptera: Thripidae) of similar age were obtained from a colony reared on orange fruits and maintained in an insectary at UWS Hawkesbury under conditions of 25±3° C., 65% RH and 16 h D:L photoperiod Young nymphs of citrus aphids Toxoptera citricida (Kirkaldy) (Hemiptera: Aphididae) were collected from lemon seedlings grown under glasshouse conditions at UWS Hawkesbury.","Workers of the termite Nasutitermes walkeri Hill (Isoptera: Termitidae) were collected from a laboratory culture at UWS Richmond which was initiated from soil, termitaria and wood on which termites were feeding were field collected at Richmond NSW, and maintained in a darkened container under conditions of 25±2° C., 35-68% RH.","Termites were fed on wood collected from near the original nest.","Moistened soil from the nest together with paper towel were placed on top of the nest and made it possible to maintain this culture for several months in the laboratory at UWS Hawkesbury.","Workers of the whitefooted house ant Technomyrmex albipes (F. Smith) (Hymenoptera: Formicidae) were field collected at UWS Hawkesbury by baiting in an empty glass jar containing sugar granules.","Pupae of housefly, Musca domestica L (Diptera: Muscidae) and different stages of American Cockroach, Periplaneta americana L (Blattodea: Blattidae) were initially supplied by C.E.R.I.T and maintained in laboratory culture at UWS Hawkesbury.","Tomato russet mites (TRM) Aculops lycopersici (Massee) (Acarina: Eriophyidae) were collected from infested tomato plants near Riverstone, NSW.","Mixed sex adults of the mosquito Culex quinquefaciatus (Diptera: Culicidae) were supplied by C.E.R.I.T, held at the in the Centre for Horticulture & Plant Science, University of Western Sydney, Richmond NSW and treated one day after arrival.","Workers of the honeybee Apis mellifera (Hymenoptera: Apidae) were collected from several field hives maintained at the apiary in the Centre for Horticulture & Plant Sciences, University of Western Sydney, Richmond NSW Adults of ash-white-flies Aleaurocanthus woglumi (Homoptera: Aleyrodidae) were field collected from an ornamental pear (Prunus sp) in the Centre for Horticulture & Plant Sciences, University of Western Sydney, Richmond NSW.","Drug store beetle Sitodrepa panicea (Coleoptera: Anobiidae) were reared on curry powder under laboratory conditions of 25±1° C. and 65±5% RH.","Mixed age groups of snails Helix apersa (Mollusca: Gastropoda) were collected from infested plants in the Centre for Horticulture & Plant Science, University of Western Sydney, Richmond NSW.","Bioassays TSM From an E. cloeziana oil extract (containing 85% tasmanone), 1.1765 g was dissolved in 5 mL ethyl alcohol and distilled water containing 200 ppm Triton X-100™ was added to prepare a 1% stock solution.","From this homogenised stock solution, further serial dilutions of 0.0125, 0.025, 0.05, 0.10, and 0.12% were prepared by mixing the required amount of stock solution in distilled water and Triton X-100 solution.","Each treatment was conducted on 60-80 TSM, which were evenly distributed on four French bean leaf discs (25 mm diam) contained in 90 mm diam.","petri dishes.","The leaf discs were placed with their underside uppermost on moist absorbent cotton wool covered with muslin netting.","Water was added to the dishes daily to prevent desiccation of the leaf discs.","Five mL aliquots (unless otherwise stated) were applied to each petri dish with a Potter precision spray tower as described by Herron et al (1995).","The average mass of solution applied to each dish was calculated to be 3.95 mg/cm2.Mortality was recorded 24 h after treatment.","Death was recognised by the absence of movement when the test organisms were mechanically stimulated.","Data were analysed using SPSS for Windows™ Version 7.Probit analysis was carried out for dose-mortality data and heterogeneity of regressions was determined by the Pearson chi-squared characteristic.","In addition, TSM was treated with E. cloeziana oil extract in combination with paraffin oil.","In particular, E. cloeziana oil extract at levels of 0.1, 0.2, 0.3, 0.4 and 0.5 g were weighed out and each was made up to a weight of 10.0 g with a formulated paraffin oil (BioPest®), which was then sonicated for 10 min.","A 1.0% v/v of each blend was prepared by mixing 1.0 mL with distilled water in a 100.0 mL volumetric flask.","TSM were transferred to the petri dish following the same standard method Five-mL aliquots were applied to each petri dish and mortality was recorded 24 h after treatment.","All blends of E. cloeziana oil extract with paraffin oil produced 100.0% mortality, compared with Biopest® alone which caused only 35.5±6.9 mortality.","The lowest concentration of E. cloeziana oil extract tested in this combination (ie.","0.01%) resulted in 100% mortality in TSM, which is significantly lower than that reported earlier in this document to produce 100% mortality with E. cloeziana oil extract alone (>0.06%).","Greenhouse Thrips The same experimental procedure used for TSM was repeated for GHT except that lemon leaf discs were used instead of French beans.","The required number of adult thrips for each treatment was transferred with a fine brush to the underside of a lemon disc of the same diameter (6 cm) as the base of a petri dish.","The lemon disc was mounted on agar with its adaxial side uppermost.","Immediately after treatment the petri dish was covered with perforated plastic wrap.","Mortality was assessed 24 h after treatment.","Tomato Russet Mite The same experimental procedure for TSM was repeated except that tomato leaf discs were used instead of French bean.","Brown Citrus Aphid Lemon leaf discs 2.5 cm diam.","were cut from tender young leaves and mounted on moistened absorbent cotton wool in 90 mm petri dish with their adaxial surface uppermost.","Each petri dish contained four leaf discs.","Uniform early instar nymphs were then transferred with a fine brush to the leaf discs (each containing 8-10 nymphs).","A Potter tower was used to apply 5 mL aliquots to each petri dish.","A control (solvent and surfactant only) was also included in the assessment.","Mortality was assessed 24 h after treatment.","Termites Twenty uniform termite workers were transferred to 90 mm petri dishes lined with the same diameter moistened filter paper (Whatman No 2).","A preliminary trial was carried out using a Potter tower to apply 5 mL aliquots of each concentration.","Using this method, all termite workers died 4 h after treatment in all concentrations, including the lowest concentration of 0.015%.","There was no mortality recorded in the blank control treatment, and all workers remained alive and active for >48 h after treatment.","Identical results were obtained from these investigations, whether the petri dishes were covered or uncovered after application of the E. cloeziana oil extract.","Subsequent investigations further assessed efficacy of E. cloeziana oil extract by releasing termite workers on fresh dried residues.","This was carried out by uniformly distributing one mL of each concentration over the entire surface area of a 90 mm diam filter paper.","When the paper was air dry, 20 termite workers were placed in each petri dish.","One hundred percent mortality was recorded in all four replicates even at the lowest concentration applied (0.015% w/v=150 ppm).","This suggests that the plant extract is a highly toxic contact poison to termites.","Ants The same experimental procedure for termites was repeated for ants, with the required number of worker ants for each treatment transferred with a fine brush to the filter paper containing a fresh dried residue of the E. cloeziana oil extract.","Immediately after release of the ants the petri dish was covered with perforated plastic wrap, which enabled any excess vapours to escape while retaining the ants.","Mortality was assessed 4 h after treatment.","In all 4 replicates 100% mortality was obtained at concentrations as low as 0.0075% w/v ai when applied at a rate of 1.0 mL to a 90 mm diam.","filter paper.","Houseflies One percent E. cloeziana oil extract was prepared using pure acetone as a diluent.","From this solution, further serial dilutions were prepared by adding the required amount in acetone.","Five mL aliquots of each concentration of E. cloeziana oil extract were dispensed into 500 mL kilner jars.","The kilner jars were immediately rotated until dryness to coat the inner surface uniformly with the E. cloeziana oil residue.","After complete dryness, 30-50 pupae were transferred to a series of clean uncovered petri dishes(45 mm diam.","), one of which was placed inside each jar.","The jar mouth was then covered with nylon netting supported by a rubber band.","All adult houseflies started to emerge from pupae after 48 h and most emerged within a 3 h period.","Flies were fed 5% sugar solution soaked in absorbent cotton wool.","Jars were maintained in an incubator at 29° C. Mortality was assessed at the end of the third day (i.e., approx.","72 h) after application of the E. cloeziana oil residues in the kilner jar and the placement of pupae inside the jars.","(This comprised 48 h for pupae to emerge and 24 h exposure to E. cloeziana oil residues which were now 48 h old).","Flies were observed to die within a few hours after emergence, whereas in the control they remained alive for >48 h after emergence.","The total number of adult houseflies that emerged in each kilner jar was counted and their mortality was recorded.","American Cockroaches Tests were conducted on 10-20 three months old nymphs (mean individual mass 0.2-0.3 g) and replicated three times.","One mL of 1.0% E. cloeziana oil extract in acetone was uniformly distributed on 90 mm diam.","Whatman No 2 filter paper.","A control treatment was also carried out using 1 mL acetone only (ie., minus E. cloeziana oil extract).","After complete dryness of the filter paper, the required number of cockroaches was transferred inside the kilner jars, and were fed dry dog food.","Kilner jar necks were covered with muslin netting supported by rubber bands.","Mortality was assessed 24 h after releasing the cockroaches, and death was recognised by the absence of movement when the test animals were mechanically stimulated.","Adult Mosquitoes A 0.3576 g of 85% ai of E. cloeziana oil extract was dissolve in pure acetone as a diluent to give approximately 0.304% concentration stock solution.","From this solution, further serial dilutions 0.152, 0.076, 0.0043 and 0.00215 were prepared by adding the required amount in acetone.","Aliquots (2.5 mL) of each concentration of E. cloeziana oil extract were dispensed into 500 mL kilner jars with total internal surface area as 286.53 cm2.The kilner jars were immediately rotated to coat the inner surface uniformly with the E. cloeziana oil extract residue, until dry.","Once completely dry, 10-25 mixed sex adult mosquitos were released into each kilner jar by allowing the mosquitoes to fly from a darkened cage into the naturally lit jars.","The jar mouth was subsequently placed on 110 mm diam.","filter paper onto which had been placed a yellow sponge soaked in 7.0% sugar solution.","Treated jars were kept under laboratory temperature and humidity conditions, (viz.","24±1° C. and 65±5% RH respectively).","Mortality was assessed 24 h after releasing the mosquitoes in the jars.","Honey Bees Ten worker honey bees, anaesthetised with carbon dioxide, were transferred to 90 mm diam.","petri dishes lined with moistened filter paper.","Five mL aliquots were applied using a Potter Spray Tower while the bees were still anaesthetised.","Yellow sponges soaked in 7% sugar solution were then placed inside petri dishes for bee sustenance, and the bees after treatment were placed in these dishes.","The lids that were perforated and covered with muslin netting were placed on the dishes, and mortality was assessed 24 hours after treatment.","Ash White Flies For each treatment, a 2.5 cm diam.","leaf disc was mounted on a moistened Whatman #2 filter paper lining the bottom of petri dishes.","Adult white flies were anaesthetised using carbon dioxide and 30-50 adults were transferred onto leaf discs.","Serial dilutions of 0.00546, 0.0220, 0.0894 & 0.1788% concentration of E. cloeziana oil extract were prepared using distilled water containing 200 ppm Triton X100.Three replicates were treated for each concentration.","Five-mL aliquots were applied to each petri dish using a Potter Spray Tower.","After treatment, petri dishes were left to dry and then covered with muslin netting.","Mortality assessment was assessed 24 h after white flies had been transferred to petri dishes.","Drug Store Beetle One mL of each concentration of E. cloeziana oil extract in pure acetone was uniformly dispensed on 90 mm diam.","Whatman No 2 filter paper.","The latter was left for 1 h to air dry before being used to line the lid of a 90 mm diam.","petri dish.","Between 10-15 mixed adults were then transferred into the petri dish, which was sealed with Parafilm™.","Mortality assessment was carried out 24 h after sealing the petri dish.","Snails Two methods were used to assess the efficacy of E. cloeziana oil extract against snails.","In the first method different age groups of adult snails were dipped directly into a solution of 0.5% concentration of E. cloeziana oil extract in distilled water containing 200 ppm Triton X10.Snails were dipped for 10 seconds and thereafter immediately filtered in a sieve.","The treated snails were divided in three 500 mL kilner jars containing French bean leaves as a food source.","The control was carried out using 0% E. cloeziana oil extract.","Mortality was assessed 24 h after treatment.","Allowing the snails to crawl on a E. cloeziana oil extract contaminated surface carried out the second method.","Two mL of 0.08% concentration of E. cloeziana oil extract in pure acetone were dispensed in 500 mL kilner jars which were rotated to uniformly cover all inner surfaces until dry.","Six different age groups of snails were put into each jar along with plant material for food and covered with muslin netting held by a rubber band.","Three replicates of each treatment (0.0 & 0.08% concentration) were carried out.","Mortality was assessed 24 h after releasing the snails inside the jars.","The above bioassay results are summarised in Table 8.TABLE 8 SUMMARY OF E. CLOEZIANA OIL EXTRACT EFFICACY AGAINST VARIOUS TARGET PESTS Method of Target Pest LD50 (95% CL) LD95 (95% CL) application Remarks TSM 0.07 0.14 Potter spray Knockdown effect (0.06-0.08) (0.12-0.16) tower; Aliquot observed 2 h after applied 2.5 mL treatment TSM <0.01 E. cloeziana oil 100% mortality at extract + paraffin 0.01% oil TRM 0.02 0.04 Potter spray tower GHT 0.10 0.125 Potter spray tower Aphid 0.08 0.30 Potter spray tower American 157.27 μg/cm2 E. cloeziana oil cockroach residues on filter nymph paper Ant <0.0075 E. cloeziana oil 100% mortality at residues on filter 0.0075% when 1 mL paper applied as residue on filter paper Termite <0.0075 E. cloeziana oil 100% mortality at residues on filter 0.0075% when 1 mL paper applied as residue on filter paper DBM 1st 0.09 0.20 5 mL Potter spray instar larva tower Housefly 69.8 μg/cm2 130.87 μg/cm2 E. cloeziana oil adult residues on kilner jar wall Adult 0.00387 0.00694 Residues on Apparently very mosquitoes (0.00328-0.00451) (0.00596-0.01051) kilner jar wall rapid knock down.","Ash white 0.01773 0.0.03728 Potter spray Assessed 4 h after fly (0.01179-0.03446) (0.02657-0.08337) tower, 2.5 mL treatment aliquot Drug store 0.03548 0.37275 Self Mortality assessed beetle contaminating by 24 h after releasing walking on filter adults.","paper Honeybees 0.12 0.40 Potter spray tower Knockdown effect Aliquot applied 5 mL observed 2 h after treatment Snails ˜0.084 E. cloeziana oil Three mL acetone residues on kilner solution for each of jar wall 3 replicates.","Snails crawled on the wall on dried residues.","Snails 100% Dipping method Three replicates mortality in 0.5% Potted Plant Investigations The efficacy of E. cloeziana oil extract against TSM was Her assessed under greenhouse conditions.","French bean plants were grown in 15 cm diam.","plastic pots in an insecticide-free glasshouse maintained at 27° C., RH 65% and natural light.","Plants were used when they reached the two true-leaf stage (ie., before trifoliate leaves appeared).","One plant was maintained in each pot (i.e., two leaves/pot).","Twenty gravid TSM females were then transferred with a fine brush to the upper surface of each leaf.","Mites were left to settle for 4 h before treatment, during which time they usually settled on the lower leaf surfaces.","E. cloeziana oil extract at a concentration of 0.07% as well as a blank control were prepared using the same procedures described above for the laboratory bioassays.","A 400 mL hand sprayer was used to apply the pesticide evenly to all aerial surfaces of the plants, to run-off.","Each treatment was replicated four times.","The mortality was assessed 24 h after treatment.","The results were recorded as mean percent mortality with standard deviation.","These results revealed that the mean percent mortality in E. cloeziana oil extract treatment and control were 92.19±6.39 and 0.25±0.46, respectively.","In summary, E. cloeziana oil extract was efficacious against all pests tested namely TSM, TRM, GHT, aphids, termites, houseflies, American cockroaches, whitefooted ants adult mosquitoes, ash whitefly, drug store beetle and snails.","It was also toxic to honeybees.","Example 8 E. cloeziana Oil Extract as a Fumigant An investigation was undertaken to determine the fumigation action of E. cloeziana oil extract against arthropods.","The test organisms used were termites, Nasutitermes walkeri.","A Whatman No 2 filter paper (90 mm diam.)","was immersed in distilled water five seconds and left to drain excess water before placing it on the bottom of a 90 mm diam.","petri dish, to provide moisture for termite workers during the experimental period.","A 1.0% E. cloeziana oil solution was prepared in pure acetone.","One mL was uniformly spread on a second filter paper, which was allowed to air dry on a sheet of aluminium foil.","It was then placed under the lid of the petri dishes containing the moist filter paper on their base.","Twenty uniform worker termites were transferred to the moist base of each test petri dish, which was subsequently covered with its lid containing the treated filter paper, and the dishes were then sealed with Parafilm™.","A similar series of control treatments was also prepared, using acetone only, for comparison.","Five replicates were used in each treatment.","The termites did not move to the dry top surface and remained on the water-moistened filter paper lining the base of the petri dishes throughout the experimental period.","Mortality was assessed 5 h after termite release.","The results revealed that E. cloeziana oil extract has highly significant fumigant effects on termites.","One hundred percent mortality was recorded in all replicates of the E. cloeziana oil treatment whereas no mortality occurred in any of the control (acetone only) replicates.","Example 9 Efficacy of Purified β-Triketones Against TSM The efficacy of purified β-triketones against TSM was investigated using the TSM bioassay described in Example 8.All β-triketones tested demonstrated a high level of activity against TSM (see Table 9).","TABLE 9 EFFICACY OF β-TRIKETONES AGAINST TSM Sample No.","wt (mg) Chemical Name LD50 and 95% CL LD95 and 95% CL 1 109.2 Agglomerone 0.15 (0.11-021) 0.33 (0.22-1.06) 2 99.5 Grandiflorone 0.04 0.13 3 165.0 Dehydroangustione 0.36 (0.33-0.41) 0.69 (0.61-0.81) 4 107.1 Angustione 0.22 (0.21-0.24) 0.35 (0.31-0.40) 5 67.9 Jensenone No direct mortality occurred in any concentration tested (0.05-0.4%) within 24 h. However, all treated TSM were unable to move normally and continued to convulse until they commenced to die 72 h after treatment.","There was no recovery.","6 110.1 Tasmanone 0.055 0.150 7 101.8 Flavesone 0.020 (0.006-0.043) 0.0876 (0.076-0.114) 8 77.0 Leptospermone 0.037 0.169 9 113.4 Isoleptospermone 0.043 (0.027-0.057) 0.071 (0.058-0.109) 10 141.1 Platyphyllol 0.070 0.23 Example 10 Efficacy of Purified β-Triketones Against GHT The efficacy of purified β-triketones against GHT was investigated using the GHT bioassay described in Example 8.All β-triketones tested demonstrated a high level of activity against GHT (see Table 9).","All β-triketones except jensenone caused 100% mortality on GHT at a concentration of 0.3% when 5 mL aliquots were applied with a Potter spray tower.","The latter β-triketone did not cause direct mortality within 24 h, but caused behavioural effects at all concentrations tested.","Convulsion and lack of movement were consistently observed and 60.0% mortality was recorded 72 h after application in the 0.4% treatment.","Examples 11 Ready-to-use miticide spray - I Ingredient Parts E. cloeziana extract 0.1 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Solvent: e.g.","ethyl alcohol, isopropyl alcohol etc 5 Tannic acid 1.0 Carrier e.g.","water 92.9 Examples 12 Concentrated natural emulsifiable concentrate spray (4.4%) - I Ingredient Parts E. cloeziana extract 4.4 Pyrethrins 7.4 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 14.7 polyoxyethylenesorbitan, organosilicate Solvent: e.g.","ethyl alcohol, isopropyl alcohol etc 73.5 Example 12 Concentrated emulsifiable concentrate spray (44%) Ingredient Parts E. cloeziana extract 22.0 Platyphyllol (natural or synthetic) 22.0 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 34.0 polyoxyethylenesorbitan, organosilicate Solvent: e.g.","ethyl alcohol, isopropyl alcohol etc 22 Example 13 Natural ready to use insecticide spray Ingredient Parts E. cloeziana extract 0.3 Lavender oil 1.0 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Solvent e.g.","ethyl alcohol, isopropyl alcohol etc 40 Carrier: Water 57.7 Example 14 Oil-based natural spray concentrate Ingredient Parts E. cloeziana extract 10.0 Petroleum oil 89.0 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Example 15 Concentrated emulsifiable concentrate spray (10%) Ingredient Parts E. cloeziana extract 10.0 Permethrin 10.0 Piperonyl butoxide 28.0 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 31.0 polyoxyethylenesorbitan, organosilicate Solvent e.g.","ethyl alcohol, isopropyl alcohol etc 21 Example 16 Molluscidal dust Ingredient Parts E. cloeziana extract 2 Anti-caking agent (e.g.","silica gel) 2 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 3 polyoxyethylenesorbitan, organosilicate Inert carrier (talc, kaolin, diatomaceous earth) 93 Example 17 Aerosol insecticidal and acaricidal spray Ingredient Parts E. cloeziana extract 1.0 Piperonyl butoxide 0.9 Propellent hydrocarbon 98.1 Example 18 Repellent Ingredient Parts E. cloeziana extract 19.5 Citronella oil 29.1 Phthalic acid dibutyl ester 29.1 N-octyl bicycloheptene dicarboxamide 22.3 Examples 19 Ready-to-use miticide spray - II Ingredient Parts Melaleuca cajeputi extract 0.2 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Solvent: e.g.","ethyl alcohol, isopropyl alcohol etc 5.0 Tannic acid 1.0 Carrier e.g.","water 92.8 Examples 20 Concentrated natural emulsifiable concentrate spray (4.4%) - II Ingredient Parts Melaleuca cajeputi extract 8.8 Pyrethrins 7.4 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 14.7 polyoxyethylenesorbitan, organosilicate Solvent: e.g.","ethyl alcohol, isopropyl alcohol etc 69.1 Examples 21 Ready-to-use miticide spray - III Ingredient Parts 99% Tasmanone 0.09 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Solvent: e.g.","ethyl alcohol, isopropyl alcohol etc 5 Tannic acid 1.0 Carrier e.g.","water 92.91 Examples 22 Concentrated natural emulsifiable concentrate spray (4.4%) - III Ingredient Parts 99% Tasmanone 4.0 Pyrethrins 7.4 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 14.7 polyoxyethylenesorbitan, organosilicate Solvent: e.g.","ethyl alcohol, isopropyl alcohol etc 73.9 Examples 23 Ready-to-use miticide spray - IV Ingredient Parts 99% Platyphyllol 0.09 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Solvent: e.g.","ethyl alcohol, isopropyl alcohol etc 5 Tannic acid 1.0 Carrier e.g.","water 92.91 Examples 24 Concentrated natural emulsifiable concentrate spray (4.4%) - IV Ingredient Parts 99% Platyphyllol 4.0 Pyrethrins 7.4 Emulsifier: e.g.","t-octylphenoxypolyethoxyethanol, 14.7 polyoxyethylenesorbitan, organosilicate Solvent: e.g.","ethyl alcohol, isopropyl alcohol etc 73.9 Example 25 Intra-Specific Crosses for Imparting Pest Resistance to a Pest Susceptible Plant Controlled and wild pollination within Eucalyptus and other important commercial Myrtaceae are thoroughly addressed in the CRC for Sustainable Production Forestry Symposium on.","Hybrid Breeding and Genetics—Controlled Pollination of Eucalypts on 12th April, 2000 and published as the proceedings of that symposium.","Genetic Pollution from Farm Forestry (Potts et al., 2001) deals more specifically with intra and inter species crosses occurring within the Myrtaceae.","Using the protocols described in the above publications, an intra-specific Eucalyptus cross breeding technique will employ the following protocol.","This begins with the selection of supreme individuals as parent stock.","Within a selected parent stock having superior pest resistant characteristics (e.g., E. cloeziana) male and female trees are identified for cross-pollination experiments.","Pollen is harvested from the male trees and either stored or directly transferred to the female trees if flowering is synchronous.","Emasculation is undertaken to preclude extraneous pollination occurring and flowers are often bagged as a further precaution.","Seed set and subsequent embryo development proceeds over the ensuing 12-24 months and F1 seeds are collected at full maturation.","Seeds are then germinated to produce seedlings that are subjected to detailed analysis to assess the transfer of traits from parent to progeny.","If the F1 progeny show a desirable mix of phenotypic traits these progeny can be used to vegetatively propagate the new variety.","If the F1 progeny show some improvement in the selected phenotypic traits, but further improvement is required selected F1 progeny can be back-crossed either within the F1 progeny or with one of the parent trees to produce an F2 progeny using the methodology outlined above.","This iterative process can be continued ad infinitum until the desired characteristics are achieved.","Example 26 Inter-Specific Crosses for Imparting Pest Resistance to a Pest Susceptible Plant Inter-species hybridisation in the wild, which is a common phenomenon within the subgenera of the Myrtaceae, have been recorded at almost 40% in the Eucalypts, 33% in Angophora 19% in Corymbia and 19% in Symphomyrtus.","Hybridisation between the major subgenera may also occur (Potts et al., 2001).","For example, Eucalyptus camaldulensis displays 14 natural hybrid crosses including E. camaldulensis XE.","robusta, E. camaldulensis X E. alba, E. camaldulensis X E. cladocalyx, E. camaldulensis X E. bigalaterita, E. camaldulensis X E. tereticornis, E. camaldulensis X E. blakelyi, E. camaldulensis X E. dwyeri, E. camaldulensis X E. rudis, E. camaldulensis X E. ovata, E. camaldulensis X E. bridgesiana, E. camaldulensis X E. viminalis, E. camaldulensis X E. largiflorens, E. camaldulensis X E. melliodora, E. camaldulensis X E. leucoxylon; 15 manipulated hybrids including E. camaldulensis X E. diversicolor, E. camaldulensis X E. grandis, E. camaldulensis X E. botryoides, E. camaldulensis X E. cladocalyx, E. camaldulensis X E. tereticornis, E. camaldulensis X E. blakelyi, E. camaldulensis X E. urophylla, E. camaldulensis X E. macarthurfi, E. camaldulensis X E. exerta, E. camaldulensis X E. maidenii, E. camaldulensis X E. viminalis, E. camaldulensis X E. globulus, F. canialdulensis X E. gunnii, E. camaldulensis X E. laevopinea, E. camaldulensis XE.","fastigata.","In another example Eucalyptus globulus displays 15 natural hybrids including E. globulus XE.","barberi, E. globulus XE.","brookeriana, E. globulus XE.","ovata, E. globulus XE.","kitsoniana, E. globulus XE.","goniocalyx, E. globulus XE.","nortonii, E. globulus X E. cypellocarpa, E. globulus X E. pseudoglobulus, E. globulus X E. bicostata, E. globulus XE.","johnstonii, E. globulus XE.","viminalis, E. globulus XE.","cordata, E. globulus XE.","rubida, E. globulus XE.","urnigera, E. globulus XE.","perriniana and 13 successful manipulated hybrids including E. globulus X E. urophylla, E. globulus X E. grandis, E. globulus XE.","robusta, E. globulus XE.","pellita, E. globulus XE.","longifolia, E. globulus X E. loxophloeba, E. globulus X E. camaldulensis, E. globulus X E. dunnii, E. globulus XE.","nitens, E. globulus XE.","maidenii, E. globulus XE.","bicostata, E. globulus X E. viminalis, E. globulus XE.","gunnii.","In another example Eucalyptus grandis displays 4 natural hybrids including E. grandis XE.","urophylla, E. grandis XE.","robusta E. grandis X E. pellita E. grandis XE.","terreticornis and 14 manipulated hybrids including E. grandis X E. urophylla E. grandis XE.","botryoides E. grandis XE.","pellita E. grandis XE.","alba E. grandis XE.","terreticornis E. grandis XE.","camaldulensis E. grandis XE.","dunnii E. nitens X E. maidenii E. grandis X E. globulus E. grandis X E. gunnii E. grandis X E. pulverulenta E. grandis X E. leucoxylon, E. grandis X E. resinifera.","In another example the Corymbia henryi/variegata/maculata/citriodora complex displays 14 natural hybrids including C. citriodora X C catenaria, C. citriodora X C. variegata, C. citriodora X C. maculata, C. maculata X C. gummifrea, C. maculata X C. intermedia, C. maculata X C. citriodora, C. maculata X C. variegata, C. variegata X C. bloxsomeri, C. variegata X C. watsoniana, C. variegata X C. citriodora, C. variegata X C. maculata, C. Henryi X C. torelliana, C henryi X C. variegata and onemanipulated hybrid C. torelliana x c. In one further example Eucalyptus cloeziana displays only 1 natural hybrid E. cloeziana X E. acmenoides.","The disclosure of every patent, patent application, and publication cited herein is hereby incorporated herein by reference in its entirety.","The citation of any reference herein should not be construed as an admission that such reference is available as “Prior Art” to the instant application.","Throughout the specification the aim has been to describe the preferred embodiments of the invention without limiting the invention to any one embodiment or specific collection of features.","Those of skill in the art will therefore appreciate that, in light of the instant disclosure, various modifications and changes can be made in the particular embodiments exemplified without departing from the scope of the present invention.","All such modifications and changes are intended to be included within the scope of the appended claims.","BIBLIOGRAPHY Bignall, C. M., Dunlop, P. J., Brophy, J. J. and Fookes, C. J. R. (1997).","Volatile Leaf Oils of some South-western and Southern Australian Species of the Genus Eucalyptus (Series I).","Part XIV.","Subgenus Monocalyptus.","Flav Frag.","Journal, 12, 177-183.Boland, D. and J. Brophy (1993).","Essential Oils of the Eucalyptus and Related Genera: Search for Chemical Trends.","Bioactive Volatile Compounds from Plants.","P, Teranishi, R. G. Buttery and H. Sugisawa.","Washington D.C., American Chemical Society.","525: 72-87.Brophy, J. J. and D. J. Boland (1990).","“Leaf Essential Oil of Two Chemotypes of Eucalyptus cloeziana F. Muell.” Journal of Essential Oil Research 2(Mar/Apr): 87-90.Brophy, J. J., R. J. Goldsack, et al.","(1995).","“Leaf Oils of the Genus Backhousia (Myrtaceae).” Journal of Essential Oil Research 7(May/Jun): 237-254.CRC for Sustainable Production Forestry.","Symposium on Hybrid Breeding and Genetics—Controlled Pollination of Eucalypts, Noosa Australia, 12th April, 2000 Hellyer, R. (1968).","“The Occurrence of O-Triketones in the Steam-Volatile Oils of some Myrtaceous Australian Plants.” Aust.","J. Chem.","21(11): 2825-2828.Herron G A, Beatie G A C, Parkes R A & Barchia I.","1995.Potter spray tower bioassay of selected citrus pests to petroleum spray oil.","Journal of Australian Entomological Society 34: 225-263.Potts, B M, Barbour, R C and Hingston, A H, 2001.Genetic Pollution from Farm Forestry.","Rural Industries Research and Development Corporation Publication (No 01/114).","Southwell, I.","A. and J. J. Brophy (2000).","“Essential oil isolates from the Australian Flora.","Part 3.” Journal of Essential Oil Research 12: 267-278."]],"string":"[\n [\n \"Methods and compositions for controlling pests\",\n \"The present invention is directed to pest-controlling compositions comprising as active ingredients one or more β-diones, particularly β-diketones and β-triketones, and to the use of these compositions inter alia for preventing, eradicating, destroying, repelling or mitigating harmful, annoying or undesired pests including insects, arachnids, helminths, molluscs, protozoa and viruses.\",\n \"The present invention is further directed to processes of preparing β-diones by de novo synthesis or from natural sources such as volatile oil-bearing plants from families including Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.\"\n ],\n [\n \"1.-24.\",\n \"(canceled) 25.A method for controlling harmful, annoying or undesired pests, other than plants, microbes, helminths and nematodes, said method comprising exposing said pests to a pest-controlling effective amount of a composition comprising a β-dione compound represented by the general formula (I) wherein A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C1, trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R&, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7; R6 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR9R10), NR9R10 or NR9OR7; R7 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R8 is selected from H, C1C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR11, SR11 or NR9R10; R9 and R10 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR12 or SR12; R11 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R12 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; X and Y are independently selected from oxygen, sulfur, —N—R4; and Q completes a 5-8-member saturated or unsaturated carbocyclic or heterocyclic ring.\",\n \"26.The method of claim 25, wherein the compound is represented by the general formula (III) wherein X, Y and Z are each independently selected from oxygen, sulfur, —N—R4 or one of C═X, C═Y or C=Z is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; C, D, E and F are each independently selected from H, C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1; and R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2 (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R1)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7.27.The method of claim 26, wherein the compound is selected from flavesone (1-isobutyroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), isoleptospermone (1-isovaleroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), leptospermone (1-valeroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), papuanone (1-pentoyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), grandiflorone (1-(2-phenylethyl)-3,3,5,5-tetramethylcyclohexan-2,4,6-trione) and jensenone (1-valeroyl-3,5-dicarbonylcyclohexan-2,4,6-trione), including analogues and derivatives thereof.\",\n \"28.The method of claim 26, wherein the compound has a structural formula selected from: 29.The method of claim 26, wherein the compound has a structural formula selected from: 30.The method of claim 26, wherein the compound has a structural formula selected from: 31.The method of claim 26, wherein the compound has a structural formula selected from: 32.The method of claim 25, wherein the compound is represented by the general formula (IV) wherein X and Y are each independently selected from oxygen, sulfur —N—R4 or one of C═X or C═Y is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; C, D, E and F are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2; and R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 cloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10, heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R1 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7.33.The method of claim 32, wherein the compound is selected from tasmanone (1-isobutroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), agglomerone (1-isobutroyl-4-methoxy-5,5-dimethylcyclohex-3-en-2,6-dione), lateriticone (1-valeroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), isolateriticone (1-isovaleroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione and platyphyllol (6,6-dimethyl-2-acetyl-5-methoxycyclohex-4-ene-1,3-dione), including analogues and derivatives thereof.\",\n \"34.The method of claim 32, wherein the compound has a structural formula selected from: 35.The method of claim 25, wherein the compound is represented by the general formula (V) wherein X and Y are independently selected from oxygen, sulfur or —N—R4; and A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl C2-C10 alkenyl, aryl or heteroaryl; C, D, E, F, G and H are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; and R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7.36.The method of claim 35, wherein the compound is selected from are angustione (1-acetyl-3,5,5-trimethylcyclohex-2,6-dione), dehydroangustione (1-acetyl-3,5,5-trimethylcyclohex-3-en-2,6-dione) and xanthostemone (1-isobutroyl-5,5-dimethylcyclohex-3-en-2,6-dione), including their analogues and derivatives.\",\n \"37.The method of claim 35, wherein the compound has a structural formula selected from: 38.The method of claim 35, wherein the compound has a structural formula selected from: 39.The method of claim 35, wherein the compound has a structural formula selected from: 40.The method of claim 25, wherein the compound is obtainable from a volatile oil-bearing organism.\",\n \"41.The method of claim 40, wherein the compound is present in an essential oil derived from said volatile oil-bearing organism.\",\n \"42.The method of claim 40, wherein the volatile oil-bearing organism is selected from volatile oil-bearing plants.\",\n \"43.The method of claim 40, wherein the volatile oil-bearing organism is selected from plants from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.\",\n \"44.The method of claim 40, wherein the volatile oil-bearing organism is selected from genera of the Myrtaceae family.\",\n \"45.The method of claim 40, wherein the volatile oil-bearing organism belongs to a genus selected from Angophora, Austromyrtus, Backhousia, Baeckea, Callistemon, Corymbia, Darwinia, Eucalyptus, Kunzea, Leptospermum, Melaleuca, Syzygium and Xanthostemon 46.The method of claim 25, wherein the pest that is controlled is selected from insects, arachnids, helminths, molluscs, protozoa and viruses.\",\n \"47.The method of claim 25, wherein the pest that is controlled is selected from insects, arachnids, helminths and molluscs.\",\n \"48.The method of claim 25, wherein the compound is used in the form of, a pest-controlling composition which comprises from about 0.00005% to about 90% by weight of said compound.\",\n \"49.-73.\",\n \"(canceled) 74.A method for controlling harmful, annoying or undesired pests, other than plants and microbes, said method comprising exposing said pests to a pest-controlling effective amount of a composition comprising a β-dione compound represented by the general formula (I) wherein A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7; R6 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR9R10), NR9R10 or NR9OR7; R7 is selected from H, C1C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R8 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR11, SR11 or NR9R10; R9 and R10 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR12 or SR12; R11 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R12 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; X and Y are independently selected from oxygen, sulfur, —N—R4; and Q completes a 5-8-member saturated or unsaturated carbocyclic or heterocyclic ring, with the proviso that when B═H1, C1-C10, alkyl or aryl, A is not CR3NR4R5 or C(R3)2OR2.75.The method of claim 74, wherein the compound is represented by the general formula (III) wherein X, Y and Z are each independently selected from oxygen, sulfur, —N—R4 or one of C═X, C═Y or C=Z is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; C, D, E and F are each independently selected from H, C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1; and R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalky, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R1)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7, with the proviso that when B═H1, C1-C10, alkyl or aryl, A is not CR3NR4R5 or C(R3)2OR2.76.The method of claim 75, wherein the compound is selected from flavesone (1-isobutyroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), isoleptospermone (1-isovaleroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), leptospermone (1-valeroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), papuanone (1-pentoyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), grandiflorone (1-(2-phenylethyl)-3,3,5,5-tetramethylcyclohexan-2,4,6-trione) and jensenone (1-valeroyl-3,5-dicarbonylcyclohexan-2,4,6-trione), including analogues and derivatives thereof.\",\n \"77.The method of claim 75, wherein the compound has a structural formula selected from: 78.The method of claim 75, wherein the compound has a structural formula selected from: 79.The method of claim 75, wherein the compound has a structural formula selected from: 80.The method of claim 75, wherein the compound has a structural formula selected from: 81.The method of claim 74, wherein the compound is represented by the general formula (IV) wherein X and Y are each independently selected from oxygen, sulfur —N—R4 or one of C═X or C═Y is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; C, D, E and F are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2; and R1 is selected from H, C1-C10 alkyl C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl.\",\n \"C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy.\",\n \"C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O2) or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7, with the proviso that when B═H1, C1-C10, alkyl or aryl, A is not CR3NR4R5 or C(R3)2OR2.82.The method of claim 81, wherein the compound is selected from tasmanone (1-isobutroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), agglomerone (1-isobutroyl-4-methoxy-5,5-dimethylcyclohex-3-en-2,6-dione), lateriticone (1-valeroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), isolateriticone (1-isovaleroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione and platyphyllol (6,6-dimethyl-2-acetyl-5-methoxycyclohex-4-ene-1,3-dione), including analogues and derivatives thereof.\",\n \"83.The method of claim 81, wherein the compound has a structural formula selected from: 84.The method of claim 74, wherein the compound is represented by the general formula (V) wherein X and Y are independently selected from oxygen, sulfur or —N—R4; and A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; C, D, E, F, G and H are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; and R1 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl, C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, c2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, c3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7, with the proviso that when B═H1, C1-C10, alkyl or aryl, A is not CR3NR4R5 or C(R3)2OR2.85.The method of claim 84, wherein the compound is selected from are angustione (1-acetyl-3,5,5-trimethylcyclohex-2,6-dione), dehydroangustione (1-acetyl-3,5,5-trimethylcyclohex-3-en-2,6-dione) and xanthostemone (1-isobutroyl-5,5-dimethylcyclohex-3-en-2,6-dione), including their analogues and derivatives.\",\n \"86.The method of claim 35, wherein the compound has a structural formula selected from: 87.The method of claim 84, wherein the compound has a structural formula selected from: 88.The method of claim 84, wherein the compound has a structural formula selected from: 89.The method of claim 74, wherein the compound is obtainable from a volatile oil-bearing organism.\",\n \"90.The method of claim 89, wherein the compound is present in an essential oil derived from said volatile oil-bearing organism.\",\n \"91.The method of claim 89, wherein the volatile oil-bearing organism is selected from volatile oil-bearing plants.\",\n \"92.The method of claim 89, wherein the volatile oil-bearing organism is selected from plants from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.\",\n \"93.The method of claim 89, wherein the volatile oil-bearing organism is selected from genera of the Myrtaceae family.\",\n \"94.The method of claim 89, wherein the volatile oil-bearing organism belongs to a genus selected from Angophora, Austromyrtus, Backhousia, Baeckea, Callistemon, Corymbia, Darwinia, Eucalyptus, Kunzea, Leptospermum, Melaleuca, Syzygium and Xanthostemon 95.The method of claim 74, wherein the pest that is controlled is selected from insects, arachnids, helminths, molluscs, protozoa and viruses.\",\n \"96.The method of claim 74, wherein the pest that is controlled is selected from insects, arachnids, helminths and molluscs.\",\n \"97.The method of claim 74, wherein the compound is used in the form of a pest-controlling composition which comprises from about 0.00005% to about 90% by weight of said compound.\",\n \"98.The method of claim 25, wherein Q completes the 5-8-member saturated or unsaturated carbocyclic or heterocyclic ring in which one or more members comprise —C(═X)—; and wherein Q is substituted with one or more substituents selected from C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1.99.The method of claim 74, wherein Q completes the 5-8-member saturated or unsaturated carbocyclic or heterocyclic ring in which one or more members comprise —C(═X)—; and wherein Q is substituted with one or more substituents selected from C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Triketones have been used for many years as herbicides for the control of undesired vegetation.\",\n \"Herbicidal triketones have been described, for example, in EP-A-338992, EP-A-336898, U.S. Pat.\",\n \"No.\",\n \"4,869,748, EP-A-186118, EP-A-186119, EP-A-186120, U.S. Pat.\",\n \"No.\",\n \"4,202,840, U.S. Pat.\",\n \"No.\",\n \"4,695,673, U.S. Pat.\",\n \"No.\",\n \"4,780,127, U.S. Pat.\",\n \"No.\",\n \"4,921,526, U.S. Pat.\",\n \"No.\",\n \"5,006,150, U.S. Pat.\",\n \"No.\",\n \"5,545,607, U.S. Pat.\",\n \"No.\",\n \"5,925,795, U.S. Pat.\",\n \"No.\",\n \"5,990,046, U.S. Pat.\",\n \"No.\",\n \"6,218,579, EP-A-249150, EP-A-137963, EP-A-394889, EP-A-506907 or EP-B-135191.Examples of herbicidal triketones are inter alia Sulcotrione (MIKADO®) whose chemical designation is 2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexandione, 2-(4-methylsulfonyloxy-2-nitrobenzoyl)-4,4,6,6-tetramethyl-1,3-cyclohexane dione; 3-(4-methylsulfonyloxy-2-nitrobenzoyl)-bicyclo-[3,2,1]octane-2,4-dione; 3-(4-methylsulfonyl-2-nitrobenzoyl)-bicyclo-[3,2,1]octane-2,4-dione; 4-(4-chloro-2-nitrobenzoyl)-2,6,6-trimethyl-2H-1,2-oxazine-3,5(4H, 6H)-dione; 3-(4-methylthio-2-nitrobenzoyl)-bicyclo[3,2,1]octane-2,4-dione; 4-(2-nitro-4-trifluoromethoxybenzoyl)-2,6,6-trimethyl-2H-1,2-oxazine-3,5(4H, 6H)-dione.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The instant invention is predicated in part on the discovery that β-diones, particularly β-diketones and β-triketones, such as those obtainable from volatile oil-bearing plants including plants from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae, exhibit significant pesticidal, especially insecticidal, arachnicidal, helminthicidal and/or molluscicidal activity.\",\n \"This discovery has been reduced to practice in novel pest-controlling compositions and methods for their preparation and use, as described hereinafter.\"\n ],\n [\n \"FIELD OF THE INVENTION THIS INVENTION relates generally to methods and compositions for controlling pests.\",\n \"More particularly, the present invention relates to pest-controlling compositions comprising as active ingredients one or more β-diones, particularly β-diketones and β-triketones, and to the use of these compositions inter alia for preventing, eradicating, destroying, repelling or mitigating harmful, annoying or undesired pests including insects, arachnids, helminths, molluscs, protozoa and viruses.\",\n \"The present invention further relates to processes of preparing β-diones by de novo synthesis or from natural sources such as volatile oil-bearing plants from families including Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.\",\n \"Bibliographic details of various publications referred to in this specification are collected at the end of the description.\",\n \"BACKGROUND OF THE INVENTION Triketones have been used for many years as herbicides for the control of undesired vegetation.\",\n \"Herbicidal triketones have been described, for example, in EP-A-338992, EP-A-336898, U.S. Pat.\",\n \"No.\",\n \"4,869,748, EP-A-186118, EP-A-186119, EP-A-186120, U.S. Pat.\",\n \"No.\",\n \"4,202,840, U.S. Pat.\",\n \"No.\",\n \"4,695,673, U.S. Pat.\",\n \"No.\",\n \"4,780,127, U.S. Pat.\",\n \"No.\",\n \"4,921,526, U.S. Pat.\",\n \"No.\",\n \"5,006,150, U.S. Pat.\",\n \"No.\",\n \"5,545,607, U.S. Pat.\",\n \"No.\",\n \"5,925,795, U.S. Pat.\",\n \"No.\",\n \"5,990,046, U.S. Pat.\",\n \"No.\",\n \"6,218,579, EP-A-249150, EP-A-137963, EP-A-394889, EP-A-506907 or EP-B-135191.Examples of herbicidal triketones are inter alia Sulcotrione (MIKADO®) whose chemical designation is 2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexandione, 2-(4-methylsulfonyloxy-2-nitrobenzoyl)-4,4,6,6-tetramethyl-1,3-cyclohexane dione; 3-(4-methylsulfonyloxy-2-nitrobenzoyl)-bicyclo-[3,2,1]octane-2,4-dione; 3-(4-methylsulfonyl-2-nitrobenzoyl)-bicyclo-[3,2,1]octane-2,4-dione; 4-(4-chloro-2-nitrobenzoyl)-2,6,6-trimethyl-2H-1,2-oxazine-3,5(4H, 6H)-dione; 3-(4-methylthio-2-nitrobenzoyl)-bicyclo[3,2,1]octane-2,4-dione; 4-(2-nitro-4-trifluoromethoxybenzoyl)-2,6,6-trimethyl-2H-1,2-oxazine-3,5(4H, 6H)-dione.\",\n \"SUMMARY OF THE INVENTION The instant invention is predicated in part on the discovery that β-diones, particularly β-diketones and β-triketones, such as those obtainable from volatile oil-bearing plants including plants from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae, exhibit significant pesticidal, especially insecticidal, arachnicidal, helminthicidal and/or molluscicidal activity.\",\n \"This discovery has been reduced to practice in novel pest-controlling compositions and methods for their preparation and use, as described hereinafter.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 shows the structures relating to the major constituents of the published Myrtaceae essential oils.\",\n \"FIG.\",\n \"2 is a representation of a GC-MS trace of E. cloeziana oil.\",\n \"FIG.\",\n \"3 is a tabular and graphical representation showing 1H NMR data recorded on a fraction (F4) obtained from silica gel chromatography of E. cloeziana oil and the structure of the major and minor isomers of the compound deduced from these data.\",\n \"FIG.\",\n \"4 is a diagrammatic representation showing various tautomeric forms of an isolated β-triketone compound in solution (CDCl3).\",\n \"DETAILED DESCRIPTION OF THE INVENTION One aspect of the present invention contemplates the use of a β-dione compound, particularly a β-diketone or a β-triketone compound, in the preparation of a composition for controlling harmful, annoying or undesired pests, said compound being represented by the general formula (I) wherein A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR5, (C—N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; R1 is selected from H, C1-C10 alkyl C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C1-C10 haloalkyl, C1-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, C1-C10 hydroxyalkyl C1-C10 thioalkyl and C1-C10 nitroalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R2 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, (CR3NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2 or NR4OR7; R3 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR4R5), NR4R5, (C═N—R4)R6, N═O, N(═O)2, NR4OR7 or SO4R7; R4 and R5 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR7 or SR7; R6 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR7, SR7, (CR8NR9R10), NR9R10 or NR9OR7; R7 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R8 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR11, SR11 or NR9R10; R9 and R10 are independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR12 or SR12; R11 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; R12 is selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl; B is H, C1-C10 alkyl, C2-C10 alkenyl, aryl or heteroaryl; X and Y are independently selected from oxygen, sulfur, —N—R4; and Q completes a 5-8-member saturated or unsaturated carbocyclic or heterocyclic ring in which optionally one or more members comprise —C(═X)—; and wherein Q is, optionally substituted with one or more substituents selected from C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1.Heterocyclic systems can be optionally attached to a moiety other than those set forth above via a carbon atom or a heteroatom of R1 to R11.Preferred compounds represented by formula (1) are β-diketones and especially preferred are β-triketones.\",\n \"As used herein, the term “alkyl” refers to linear or branched chains.\",\n \"The term “haloalkyl” refers to an alkyl group substituted by at least one halogen.\",\n \"Similarly the term “haloalkoxy” refers to an alkoxy group substituted by at least one halogen.\",\n \"As used herein the term “halogen” refers to fluorine, chlorine, bromine and iodine.\",\n \"As used herein the term “aryl” refers to aromatic carbocyclic ring systems such as phenyl or naphthyl, anthracenyl, especially phenyl.\",\n \"Suitably, aryl is C6-C14 with mono, di, tri, tetra and penta substitution containing OR2, F, Cl, Br, I, NO2, CF3, COR1, NR4R5, SO2R2, SR2.As used herein the terms “heterocycle”, “heterocyclic”, “heterocyclic systems” and the like refer to a saturated, unsaturated, or aromatic carbocyclic group having a single ring, multiple fused rings (for example, bicyclic, tricyclic, or other similar bridged ring systems or substituents), or multiple condensed rings, and having at least one heteroatom such as nitrogen, oxygen, or sulfur within at least one of the rings.\",\n \"This term also includes “heteroaryl” which refers to a heterocycle in which at least one ring is aromatic.\",\n \"Any heterocyclic or heteroaryl group can be unsubstituted or optionally substituted with one or more groups, as defined above.\",\n \"Further, bi- or tricyclic heteroaryl moieties may comprise at least one ring, which is either completely, or partially, saturated.\",\n \"Suitable heteroaryl moieties include, but are not limited to oxazolyl, thiazaoyl, thienyl, furyl, 1-isobenzofuranyl, 2H-pyrrolyl, N-pyrrolyl, imidazolyl, pyrazolyl, isothiazolyl, isooxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyradazinyl, indolizinyl, isoindolyl, indoyl, indolyl, purinyl, phthalazinyl.\",\n \"The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.\",\n \"By way of example, “an element” means one element or more than one element.\",\n \"A preferred carbocyclic ring formed by Q is an optionally substituted cyclohexanedione.\",\n \"A preferred subgroup of compounds of formula (1) is represented by formula (II) Such compounds may exist in a number of tautomeric forms.\",\n \"For example, in the case wherein X and Y are each oxygen, and B is hydrogen, then the compounds of formula II may exist as one or more of the structural formulae shown below.\",\n \"It is intended that all such tautomeric structures are included within the scope of the present invention.\",\n \"It should also be appreciated that some of the compounds of formula (I) are capable of existing as different geometric isomers and diastereomers.\",\n \"The invention thus includes both the individual isomers and mixtures of such isomers.\",\n \"Another preferred subgroup of compounds of formula (I) is represented by formula (II) wherein X, Y and Z are each independently selected from oxygen, sulfur, —N—R4 or one of C═X, C═Y or C=Z is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is as defined above; C, D, E and F are each independently selected from H, C1-C10 alkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2, C2-C10 1-arylalkyl, C2-C10 2-arylalkyl or (C═X)R1; and R1, R2, R2, R4 and R5 are as defined above.\",\n \"Preferred β-diones represented by formula (III) are flavesone (1-isobutyroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), isoleptospermone (1-isovaleroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), leptospermone (1-valeroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), papuanone (1-pentoyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), grandiflorone (1-(2-phenylethyl)-3,3,5,5-tetramethylcyclohexan-2,4,6-trione) and jensenone (1-valeroyl-3,5-dicarbonylcyclohexan-2,4,6-trione), including analogues and derivatives thereof.\",\n \"By way of example, flavesone analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of flavesone is shown for comparative purposes: Non-limiting examples of isoleptospermone analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of isoleptospermone is shown for comparative purposes: Non-limiting examples of leptospermone analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of leptospermone is shown for comparative purposes: Non-limiting examples of jensenone analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of jensenone is shown for comparative purposes: Another preferred subgroup of compounds of formula (1) is represented by formula (IV) wherein X and Y are each independently selected from oxygen, sulfur —N—R4 or one of C═X or C—Y is CH2; A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, N4, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is as defined above; C, D, E and F are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2, SO4R2; and R1, R2, R2, R4 and R5 are as defined above.\",\n \"Preferred β-diones represented by formula (IV) are tasmanone (1-isobutroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), agglomerone (1-isobutroyl-4-methoxy-5,5-dimethylcyclohex-3-en-2,6-dione), lateriticone (1-valeroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione), isolateriticone (1-isovaleroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione and platyphyllol (6,6-dimethyl-2-acetyl-5-methoxycyclohex-4-ene-1,3-dione), including analogues and derivatives thereof.\",\n \"Non-limiting examples of tasmanone analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of tasmanone is shown for comparative purposes: Another preferred subgroup of compounds of formula (I) is represented by formula (V) wherein X and Y are independently selected from oxygen, sulfur or —N—R4; and A is (C═O)R1, (C═S)R1, OR2, SR2, (CR3NR4R5), C(R3)2OR2, NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; B is as defined above; C, D, E, F, G and H are each independently selected from H, C1-C10 alkyl, C2-C10 arylalkyl, C3-C6 cycloalkyl, C2-C10 alkenyl, C2-C10 heteroarylalkyl, C2-C10 haloalkyl, C2-C10 dihaloalkyl, C2-C10 trihaloalkyl, C2-C10 haloalkoxy, OR2, SR2, (CR3NR4R5), NR4R5, (C═N—R4)R1, N═O, N(═O)2, NR4OR2 or SO4R2; and R1, R2, R2, R4 and R5 are as defined above.\",\n \"More specifically unsaturation, epoxides and thioexpoxides may exist at positions designated by H (or G) connected to F (or E) or F (or E) connected to C (or D).\",\n \"A four-membered ring forming a part of a bicyclic structure may exist at positions designated by H (or G) connected to C (or D).\",\n \"Preferred β-diones represented by formula (V) are angustione (1-acetyl-3,5,5-trimethylcyclohex-2,6-dione), dehydroangustione (1-acetyl-3,5,5-trimethylcyclohex-3-en-2,6-dione) and xanthostemone (1-isobutroyl-5,5-dimethylcyclohex-3-en-2,6-dione), including their analogues and derivatives.\",\n \"By way of example, angustione analogues contemplated by the present invention include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of angustione is shown for comparative purposes: Non-limiting examples of dehydroangustione analogues include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of dehydroangustione is shown for comparative purposes: Non-limiting examples of xanthostemone analogues include, but are not restricted to, compounds having the following structural formulae, wherein the structural formula of xanthostemone is shown for comparative purposes: Derivatives of the above compounds include, but are not restricted to, ethoxylate derivatives, propoxylate derivatives, hydrates, aldehyde derivatives, ester derivatives, ether derivatives, alcohol derivatives, phenol derivatives, amine derivatives, other biologically or chemically equivalent substances, and any combination of two or more of the foregoing.\",\n \"Similarly effective as pesticides are salts of the above compounds, including mono-valent salts (e.g., sodium, potassium) and di-valent metal salts (e.g.\",\n \"calcium, magnesium, iron or copper) and ammonium salts (e.g., isopropyl ammonium, trialkyl and tetraalkylammonium salts).\",\n \"The compounds according to any one of formulae (I)-(V) can be prepared according to methods analogous to those known in the art for the preparation of β-diones.\",\n \"Exemplary methods are disclosed for example in EP-A-338992, EP-A-336898, U.S. Pat.\",\n \"No.\",\n \"4,202,840, U.S. Pat.\",\n \"No.\",\n \"4,869,748, EP-A-186118, EP-A-186119, EP-A-186120, U.S. Pat.\",\n \"No.\",\n \"4,695,673, U.S. Pat.\",\n \"No.\",\n \"4,780,127, U.S. Pat.\",\n \"No.\",\n \"4,921,526, U.S. Pat.\",\n \"No.\",\n \"5,006,150, U.S. Pat.\",\n \"No.\",\n \"5,545,607, U.S. Pat.\",\n \"No.\",\n \"5,925,795, U.S. Pat.\",\n \"No.\",\n \"5,990,046, U.S. Pat.\",\n \"No.\",\n \"6,218,579, EP-A-249150, EP-A-137963, EP-A-394889 EP-A-506907 or EP-B-135191.More particularly, compounds according to formulae (III)-(V) can be synthesised using the representative procedures outlined below.\",\n \"For compounds according to formula (III), 1,3,5-trihydroxybenzene 1 is reacted with CH3CN in the presence of zinc chloride and hydrochloric acid, according to A. H. Blatt (1943, Org.\",\n \"Synth.\",\n \"Col. II, 522-523), affording 1-acetyl-2,4,6-trihydroxybenzene 2 (phloroacetophenone) (R=Me) (Scheme 1).\",\n \"R groups other than methyl are depicted above.\",\n \"Reaction of 1-acetyl-2,4,6-trihydroxybenzene 2 affording 1-acetyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione 3 is a representative procedure for all compounds according to formula (III) (R. A.\",\n \"Gray et al., U.S. Pat.\",\n \"No.\",\n \"4,202,840) (Scheme 2).\",\n \"Anhydrous MeI (6eq) is slowly added, at room temperature under an atmosphere of nitrogen, to a mechanically stirred solution of 1-acetyl-2,4,6-trihydroxybenzene 2 (1 eq) and sodium ethoxide (6eq) in anhydrous methanol.\",\n \"The mixture is refluxed for 4 hours.\",\n \"On cooling the mixture is concentrated under vacuo, providing a residue, which is diluted with water and acidified with 2 M hydrochloric acid.\",\n \"Diethylether extracts are washed with saturated sodium sulfite solution, water and then dried (Na2SO4).\",\n \"Evaporation of the diethylether provides the desired product 3.For mono, di, tri and tetra B, C, D, E, F substitution patterns, reactions between one and seven mole equivalents of R—I and sodium ethoxide are used.\",\n \"Lawasson's reagent is used for conversion of oxygen into sulfur groups and sodium borohydride or sodium cyanoborohydride is used to reduce ketone, thioketone and imino groups.\",\n \"When additional carbonyl groups are introduced into the cyclohexane ring system the procedure of Crow is utilised (M. L. Bolte et al., 1985, Agric.\",\n \"Biol.\",\n \"Chem., 49, 761).\",\n \"Compounds of formula (IV) can be prepared according to a first representative procedure, as follows: 3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 4 (1 mole eq), prepared according to Herzig (J. Herzig, and F. Wenzel, Monatsh, 1903, 24, 101), is dissolved in anhydrous diethylether and hexamethylphosphoramide (solvent ratio, 20:1 respectively) under an atmosphere of nitrogen.\",\n \"The mixture is cooled to 0° C. and lithium hydride (1.1 mole eq) (60% in mineral oil) is added in portions.\",\n \"After addition the mixture is stirred for a further 10 mins before the addition of benzoyl cyanide 5 [R—CO—CN, R is depicted above] (1.1 mole eq).\",\n \"The mixture is allowed to warm to room temperature over 12 h at which time the reaction is quenched with water and partitioned.\",\n \"The ether layer is dried (Na2SO4) and evaporated affording crude 1-benzoyl-3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 6 which is purified by SiO2 column chromatography (hexane/ethyl acetate, gradient) (Scheme 3).\",\n \"Alternatively, compounds of formula (IV) can be prepared according to a second representative procedure, as follows: 3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 4 (1 mole eq) (commercially available) and benzoyl cyanide are dissolved in anhydrous dichloromethane and cooled to 0° C. under an atmosphere of nitrogen.\",\n \"To the cooled solution is added anhydrous finely powdered zinc chloride (1.1 mole eq.)\",\n \"followed by slow addition of triethylamine (1.2 mole eq).\",\n \"The reaction mixture is stirred at room temperature for 5-6 h and then poured into 2 M hydrochloric acid.\",\n \"The mixture is partitioned and the dichloromethane layer is washed with 5% sodium carbonate.\",\n \"The aqueous carbonate phase is then acidified with hydrochloric acid and extracted with methylene chloride and dried (Na2SO4).\",\n \"The solvent is removed and the residue subjected to SiO2 column chromatography (hexane/ethyl acetate) affording 1-benzoyl-3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 6 (W. J. Michaely and G. W. Kraatz, EP-B-135191).\",\n \"Compounds of formula (V) can be prepared according to a first representative procedure, as follows: 4,4-dimethylcyclohexane-1,3-dione 7 (1 mole eq) (commercially available) is dissolved in anhydrous diethylether and hexamethylphosphoramide (solvent ratio, 20:1 respectively) under an atmosphere of nitrogen.\",\n \"The mixture is cooled to 0° C. and lithium diisopropylamide (2.1 mole eq) is added dropwise over 40 mins.\",\n \"The mixture is then stirred for a further 10 mins before the addition of methyl iodide (1 mole eq).\",\n \"The mixture is stirred for 12 h and then benzoyl cyanide (2 mole eq) is added and the mixture stirred for a further 24 h. The reaction was quenched with water and the ether layer partitioned and dried (Na2SO4).\",\n \"The solvent was removed and the residue subjected to SiO2 column chromatography (hexane/ethyl acetate) affording 1-benzoyl-3,3,5-trimethylcyclohexan-2,6-dione 8 (Scheme 4).\",\n \"Alternatively, the compounds of formula (V) can be prepared according to a second representative procedure, as follows: 4,4-dimethyl-1,3-cyclohexanedione 7 (1 mole eq) (commercially available) and benzoyl cyanide are dissolved in anhydrous dichloromethane and cooled to 0° C. under an atmosphere of nitrogen.\",\n \"To the cooled solution is added anhydrous finely powdered zinc chloride (1.1 mole eq) followed by slow addition of triethylamine (1.2 mole eq).\",\n \"The reaction mixture is stirred at room temperature for 5-6 h and then poured into 2 M hydrochloric acid.\",\n \"The mixture is partitioned and the dichloromethane layer washed with 5% sodium carbonate.\",\n \"The aqueous carbonate phase is then acidified with hydrochloric acid and extracted with methylene chloride and dried (Na2SO4).\",\n \"The solvent is removed and the residue subjected to SiO2 column chromatography (hexane/ethyl acetate) affording 1-benzoyl-3,3,5-trimethylcyclohexan-2,6-dione 8.(W.\",\n \"J. Michaely and G. W. Kraatz, EP-B-135191).\",\n \"Dehydroangustione and xanthostemone derivatives are simply derived from dehydrogenation of angustione derivatives, for example, by treatment of 1-benzoyl-3,3,5-trimethylcyclohexan-2,6-dione 8 with palladium on charcoal in methanol, which thereby affords 1-benzoyl-3,5,5-trimethylcyclohex-3-en-2,6-dione 9 (Scheme 5).\",\n \"Metal salts (enolates) of the above compounds can be prepared by the reaction of triketone derivatives with the corresponding metal hydroxides suspended in methanol or ethanol.\",\n \"Trialkylammonium salts can be prepared by the reaction of triketone derivatives (e.g., 3) with trialkylamines in a chlorinated solvent such as dichloromethane.\",\n \"Tetraalkylammonium salts can be prepared by adding a halogenated tetraalkylammonium salt to a metal salt in dichloromethane, which precipitates the metal halide removed by filtration.\",\n \"The pure material is obtained by evaporation of the filtrate.\",\n \"The present inventors have discovered that the β-diones of the invention can be obtained from natural sources and, in particular, from volatile oil-bearing organisms.\",\n \"Accordingly, in another aspect, the present invention encompasses the use of a β-dione compound, particularly a β-diketone or a β-triketone compound, obtainable from a volatile oil-bearing organism, including an analogue or derivative thereof, in the preparation of a pesticidal composition for controlling harmful, annoying or undesired pests.\",\n \"The present invention contemplates the use of any volatile oil-bearing organism that produces β-diones, preferably the β-diones according to any one of formulae (I)-(V), and especially β-diketones and/or β-triketones, for the preparation of the pesticidal compositions of the invention.\",\n \"Preferred volatile oil-bearing organisms are volatile oil-bearing plants including, but not restricted to, plants from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.\",\n \"Preferably, the volatile oil-bearing plant is selected from genera of the Myrtaceae family including, but not limited to, Angophora, Austromyrtus, Backhousia, Baeckea, Callistemon, Corymbia, Darwinia, Eucalyptus, Kunzea, Leptospermum, Melaleuca, Syzygium and Xanthostemon.\",\n \"Thus, the compositions of the present invention may contain as active ingredients substantially purified β-diones or crude β-dione-containing extracts obtained from a volatile oil-bearing organism, preferably a volatile oil-bearing plant.\",\n \"Volatile oils, also known in the art as essential oils, typically comprise a volatile mixture of esters, aldehydes, alcohols, ketones and terpenes, which can be prepared from botanical materials or plant cell biomass from cell culture.\",\n \"Volatile oils can be prepared by subjecting botanical materials to a distillation process, for example.\",\n \"A number of different procedures can be used for distillation.\",\n \"For example, plant matter (e.g., foliage, stems, roots, seeds, bark etc) of a volatile oil-bearing plant is placed in a suitable still and steam distillation is used to break down the cells of the plant to release the oil.\",\n \"The steam is then condensed and the oil phase is separated from the aqueous phase to obtain the volatile oil.\",\n \"It will be appreciated that other methods of volatile oil extraction (e.g., solvent extraction) are known to those of skill in the art and it will be understood, in this regard, that the present invention is not limited to the use or practice of any one particular method of extracting volatile oils.\",\n \"Suitably, the compositions comprise naturally-occurring compounds derived from a volatile oil-bearing organism.\",\n \"Thus, in a preferred embodiment, the pesticidal composition of the invention comprises one or more β-dione active compounds, particularly β-diketone- and/or β-triketone-active compounds, that are derived from the volatile oil of a volatile oil-bearing organism.\",\n \"In this embodiment, the composition may optionally contain a naturally-occurring carrier and/or other naturally-occurring additives.\",\n \"Naturally-occurring additives contemplated by the present invention include natural antioxidants, which can be used advantageously to reduce the effect of oxidation of the compounds of the invention.\",\n \"An example of a suitable naturally-occurring antioxidant is α-tocopherol.\",\n \"Other additives, such as naturally-occurring stabilisers, are also contemplated, which may desirably be added to improve the stability and shelf life of the composition.\",\n \"Examples of suitable natural stabilisers include gum arabic, guar gum, sodium caseinate, polyvinyl alcohol, locust bean gum, xanthan gum, kelgum, and mixtures thereof.\",\n \"In an alternate embodiment, the naturally-occurring compounds derived from a volatile oil may be modified or derivatised to improve, for instance, their shelf-life, stability, activity and/or bioavailability.\",\n \"The compounds of the present invention are useful for controlling harmful, annoying or undesired pests.\",\n \"They may be used singularly or in combination with other pest-controlling compounds of the invention.\",\n \"By “controlling” is meant preventing, combating, eradicating, destroying, repelling, or mitigating pests or increasing the mortality or inhibiting the growth and/or development of pests.\",\n \"The term “pest” is used herein in its broadest sense and includes within its scope insects, arachnids (e.g.\",\n \"acari, spiders), helminths (e.g., nematodes), molluscs, protozoa (e.g., Plasmodium sp.\",\n \"Paramecium sp.\",\n \"), viruses (e.g., herpesviruses) and the like.\",\n \"Suitable applications for such control include, but are not limited to, combating and/or eradicating infestations by pests in animals (including humans) and/or plants (including trees) and/or stored products, which includes the administration to the animal or site of an effective quantity of a compound of the invention.\",\n \"By “effective amount” is meant the administration or application of that amount of active compound, either in a single dose or as part of a series, that is effective for controlling a significant number of pests.\",\n \"Thus, for example, a “pesticidally-effective” amount is the amount of active compound that is effective for increasing the mortality or decreasing the growth of a significant number of pests.\",\n \"Alternatively, a “pest-repelling” effective amount is the amount of active compound that is noxious to, and/or induces behavioural changes in, a significant number of pests.\",\n \"The effective amount will vary depending upon the taxonomic group of pest exposed to the active compound, the formulation of the composition, and other relevant factors.\",\n \"It is expected that the amount will fall in a relatively broad range that can be determined through routine trials.\",\n \"Accordingly, the compounds of the present invention can be used as pesticides, such as but not limited to insecticides, arachnicides, anti-helminthics, molluscicides antivirals, antiprotozoals and the like, or as pest repellents including repellents of insects, arachnids, helminths, molluscs, protozoa and viruses.\",\n \"In especially preferred embodiments, the compounds of the present invention are used in the control of insects, arachnids, helminths or molluscs.\",\n \"In practice, the compounds can be applied as formulations containing the various adjuvants and carriers known to or used in the industry for facilitating bioavailability, stability and dispersion.\",\n \"The choice of formulation and mode of application for any given compound may affect its activity, and selection will be made accordingly.\",\n \"In general, a pest-controlling compound of the invention can be compounded with appropriate inert carriers and additives in an appropriate ratio by means of dissolving, separating, suspending, mixing, impregnating, adsorbing or precipitating operation to formulate into oil formulations, emulsifiable concentrates, wettable powders, flowables, granules, powders, dusts, solutions, suspensions, emulsions, controlled-release forms such as microcapsules, aerosols or fumigants.\",\n \"Typically, the compounds of the present invention can be mixed with a solid carrier, liquid carrier or gas carrier, optionally together with a surfactant and other adjuvants useful for such formulations.\",\n \"The compounds of the invention can be used in an amount from about 0.00005% to about 90% by weight as contained in these formulations as their active component.\",\n \"As used herein, the term “about” refers to a quantity, level, value or amount that varies by as much as 30%, preferably by as much as 20%, and more preferably by as much as 10% to a reference quantity, level, value or amount.\",\n \"Where the compounds are in the form of β-dione-containing extracts, the formulations will usually comprise as their principal active ingredient from about 0.0001% to about 90%, preferably from about 0.0001% to about 50%, more preferably from about 0.0005% to about 10%, even more preferably from about 0.0005% to about 5%, even more preferably from about 0.0005% to about 1% and still even more preferably from about 0.001% to about 0.5% by weight of the extract.\",\n \"Alternatively, where the compounds are in the form of substantially purified preparation of β-diones, the formulations will usually comprise as their principal active ingredient from about 0.00005% to about 90%, preferably from about 0.0001% to about 50%, more preferably from about 0.0005% to about 10%, even more preferably from about 0.001% to about 5% and still even more preferably from about 0.001% to about 1% by weight of the substantially purified β-dione.\",\n \"By “substantially purified” is meant a compound which has been separated from components that naturally accompany it.\",\n \"Typically, a compound is substantially pure when at least 60%, more preferably at least 75%, more preferably at least 90%, and most preferably at least 99% of the total material (by volume, by wet or dry weight, or by mole percent or mole fraction) in a sample is the compound of interest.\",\n \"Purity can be measured by any appropriate method, e.g., by chromatography or HPLC analysis.\",\n \"Examples of solid carriers useful in preparing the formulations are clays including kaolin clay, diatomite, water-containing synthetic silicon oxide, bentonite, Fubasami clay, and acid clay; talcs; ceramics; inorganic minerals such as Celite, quartz, sulfur, active carbon, calcium carbonate and hydrated silica; and chemical fertilisers such as ammonium sulfate, ammonium phosphate, ammonium nitrate, urea and ammonium chloride, these solid carriers being finely divided or granular.\",\n \"Examples of useful liquid carriers are water, alcohols such as methanol and ethanol, ketones such as acetone and methyl ethyl ketone, aromatic hydrocarbons such as benzene, toluene, xylene, ethylbenzene and methylnaphthalene, aliphatic hydrocarbons such as hexane, cyclohexane, kerosene and light oil, esters such as ethyl acetate and butyl acetate, nitriles such as acetonitrile and isobutyronitrile, ethers such as diisopropyl and dioxane, acid amides such as N,N-dimethylformamide and N,N-dimethylacetamide, halogenated hydrocarbons such as dichloromethane, trichloroethane and carbon tetrachloride, dimethyl sulfoxide, and fish oils, mineral oils, plant derived oils such as canola oil, cotton-seed oil, soybean oil and sesame oil as well as essential oils such as lavender oil, eucalyptus oil, tea tree oil, citrus oil etc.\",\n \"Solid or liquid carriers can be used alone or in combination.\",\n \"Examples of gas carriers, i.e., those of propellants, are butane gas, LPG (liquefied petroleum gas), dimethyl ether, fluorocarbons and carbon dioxide gas.\",\n \"Examples of surfactants are alkylsulfuric acid esters, alkylsulfonic acid salts, alkylarylsulfonic acid salts, alkyl aryl ethers and polyoxyethylene adducts thereof, polyethylene glycol ethers, polyhydric alcohol esters, sugar alcohol derivatives, sorbitane monolaurate, alkylallyl sorbitane monolaurate, alkylbenzene sulfonate, alkylnaphthalene sulfonate, lignin sulfonate, and sulfuric acid ester salts of higher alcohols.\",\n \"These surfactants may be used alone or in combination.\",\n \"Examples of adjuvants for the formulations, such as binders and dispersants, are casein, gelatin, polysaccharides such as starch, gum arabic, cellulose derivatives and alginic acid, lignin derivatives, bentonite, sugars and water-soluble synthetic high-molecular-weight substances such as polyvinyl alcohol, polyvinyl pyrrolidone and polyacrylic acids.\",\n \"Examples of stabilisers are PAP (acid isopropyl phosphate), BHT (2,6-di-tert-butyl-4-methylphenol), BHA (mixture of 2-tert-butyl-4-methoxyphenol and 3-tert-butyl-4-methoxyphenol), synergists such as piperonyl butoxide, vegetable oils, mineral oils, fish oils, surfactants and fatty acids or esters thereof.\",\n \"Emulsifying agents that may be used are suitably one or more of those selected from non-ionic or anionic emulsifying agents.\",\n \"Examples of non-ionic emulsifying agents include, but are not restricted to, polyoxyethylenealkylphenylether, polyoxyethylenealkylether, polyethyleneglycol fatty ester, sorbitan fatty ester, polyoxyethylene sorbitan fatty ester, polyoxyethylenesorbitol fatty ester, polyoxyethylenepolyoxypropylenealkylether.\",\n \"Examples of anionic emulsifying agents include alkyl sulphates, polyoxyethylenealkylether sulphates, sulfosuccinates, taurine derivatives, sarcosine derivatives, phosphoric esters, alkylbenzenesulfonates and the like.\",\n \"A mixture consisting of polyoxyethylenestyrylphenylether and calcium allylbenzenesulfonate is preferred.\",\n \"These emulsifying agents may be used in an amount of 5 to 20 weight parts per 100 weight parts of the compositions of the present invention.\",\n \"Formulation thus obtained can be used solus or diluted, for example, with water or other diluent.\",\n \"The formulations can be used also as admixtures with other pesticides such as insecticides, arachnids, anti-helminthics, molluscicides, herbicides, plant growth regulators, synergists, soil improvers, baits and the like, or can be used simultaneously with such agents without mixing.\",\n \"For example, the pest-controlling compounds of the invention can be combined with other naturally derived bioactive compounds or extracts such as neem or its components, derris, pyrethrum; microbial extracts such as avermectins or streptomycins; with synthetic insecticides, acaricides, molluscicides, anti-helminthics; anti-protozoals, antivirals or with microorganisms having insecticidal, acaricidal, molluscicidal, anti-helminthic anti-protozoal or antiviral activity e.g., bacteria such as Bacillus thuringiensis, Bacillus popillae, entomogenous fungi such as Metarhizium spp., Verticillium lecanii, nematodes such as Steinernema spp and Heterorhabditis.\",\n \"Alternatively, or in addition, the pest-controlling compounds of the invention can be combined with synergists such as piperonyl butoxide, and with ultraviolet screening compounds of natural or synthetic origin.\",\n \"When used as an agricultural pesticide, the compound of the invention is preferably applied usually in an amount of 0.01 to 500 g/100 m2.When an emulsifiable concentrate, wettable powder or flowables are used as diluted with water, the compound is applied usually at a concentration of 0.1 to 1000 ppm, preferably 1 to 500 ppm.\",\n \"The granular or dust can be applied without dilution.\",\n \"The amount or concentration of application, although exemplified above, can be suitably increased or reduced according to the type of preparation, time, place, method of application, kind of pest and extent of harm or annoyance suffered.\",\n \"The invention also contemplates the use of the above described β-dione compounds in pest repellent, particularly insect repellent, compositions.\",\n \"Repellent compositions contemplated by the present invention include those that are noxious to, and/or induce behavioural changes in, a pest.\",\n \"The latter compositions suitably comprise an activity including, but not restricted to, an antifeedant activity, an oviposition deterrent activity and an insect growth regulatory activity.\",\n \"Insect repellent compositions in various dosage forms can be prepared by blending the above-described β-dione compounds as active ingredients with a base of cosmetics or pharmaceuticals, which are usually applied to human bodies or animals.\",\n \"They can be formulated in, for example, lotions, aerosols, milky lotions, creams or the like.\",\n \"These compounds can be further incorporated with other insect repellents, antioxidants, UV-absorbers, humectants or other additives.\",\n \"The above compounds or the above-prepared compositions of the present invention can be applied directly to human bodies or animals.\",\n \"Besides, substrates, such as sheets, films, nets, timber or the like, which have preliminarily been treated with the above compounds or compositions by means of application, impregnation or blending, can also be used.\",\n \"The quantity of the above compounds to be formulated in the noxious-insect repellents depends upon the dosage form, usage or other conditions.\",\n \"Suitable dosages may be selected from about 0.1% to about 90% by weight.\",\n \"Thus, in another aspect of the present invention there is provided a method for controlling harmful, annoying or undesired pests, said method comprising exposing said pests to a pest-controlling effective amount of a composition comprising a β-dione compound as broadly described above.\",\n \"Preferred embodiments of this type include exposing said pests to a pesticidally effective amount or a pest-repelling effective amount of said composition.\",\n \"The terms “comprise”, “comprises” and “comprising” and the like refer, unless the context requires otherwise, to the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements.\",\n \"The compositions and methods of the present invention may be applied to pests including, but not restricted to, insects, arachnids, helminths, molluscs, protozoa and viruses.\",\n \"For example, suitable insects that fall within the scope of the present invention include those: (a) from the order of the lepidopterans (Lepidoptera), for example, Adoxophyes orana, Agrotis upsilon, Agrotis segetum, Alabama argillacea, Anticarsia gemmatalis, Argyresthia conjugella, Autographa gamma, Cacoecia murinana, Capua reticulana, Choristoneuraffumiferana, Chilo partellus, Choristoneura occidentalis, Cirphis unipuncta, Cnaphalocrocis medinalis, Crocidolomia binotalis, Cydia pomonella, Dendrolimus pini, Diaphania nitidalis, Diatraea grandiosella, Earias insulana, Elasmopalpus lignosellus, Eupoecilia ambiguella, Feltia subterranea, Grapholitha funebrana, Grapholitha molesta, Heliothis armigera, Heliothis virescens, Heliothis zea, Hellula undalis, Hibernia defoliaria, Hyphantria cunea, Hyponzomeuta malinellus, Keiferia lycopersicella, Lambdina fiscellaria, Laphygma exigua, Leucoptera scitella, Lithocolletis blancardella, Lobesia botrana, Loxostege sticticalis, Lymnantria dispar, Lymantria monacha, Lyonetia clerkella, Manduca sexta, Malacosoma neustria, Mamestra brassicae, Mocis repanda, Operophthera brumata, Orgyia pseudotsugata, Ostrinia nubilalis, Pandemis heparana, Panolisflammea, Pectinophora gossypiella, Phthorimaea operculella, Phyllocnistis citrella, Pieris brassicae, Plathypena scabra, Platynota stultana, Plutella xylostella, Prays citri, Prays oleae, Prodenia sunia, Prodenia ornithogalli, Pseudoplusia includens, Rhyacionia fiustrana, Scrobipalpula absoluta, Sesamiainferens, Sparganothis pilleriana, Spodoptera fugiperda, Spodoptera littoralis, Spodoptera litura, Syllepta derogata, Synanthedon myopaeformis, Thaumatopoea pityocampa, Tortrix viridana, Trichoplusia ni, Tryporyza izcertulas, Zeiraphera canadensis, (b) furthermore Galleria mellonella and Sitotroga cerealella, Ephestia cautella, Tineola bisselliella; (c) from the order of the beetles (Coleoptera), for example, Anthonomus grandis, Anthonomus pomorum, Apion vorax, Atomaria linearis, Blastophagus piniperda, Cassida nebulosa, Cerotoma trifurcata, Ceuthorhynchus assimilis, Ceuthorhynchus napi, Chaetocnema tibialis, Conoderus vespertinus, Crioceris asparagi, Dendroctonus refipennis, Diabrotica longicornis, Diabrotica 12-punctata, Diabrotica virgifera, Epilachna varivestis, Epitrix hirtipennis, Eutinobothrus brasiliensis, Hylobius abietis, Hypera brunneipennis, Hypera postica, Ips typographus, Lema bilineata, Lema melanopus, Leptinotarsa decemlineata, Limonius californicus, Lissorhoptrus oryzophilus, Melanotus communis, Meligethes aeneus, Melolontha hippocastani, Melolontha melolontha, Oulema oryzae, Ortiorrhynchus sulcatus, Otiorrhynchus ovatus, Phaedon cochleariae, Phyllopertha horticola, Phyllophaga sp., Phyllotreta chrysocephala, Phyllotreta nemorum, Phyllotreta striolata, Popillia japonica, Psylliodes napi, Scolytus intricatus, Sitona lineatus, (d) furthermore Bruchus rufimanus, Bruchus pisorum, Bruchus lentis, Sitophilus granaria, Lasioderna serricorne, Oryzaephilus surinamensis, Rhyzopertha dominica, Sitophilus oryzae, Tribolium castaneum, Trogodernia granarium, Zabrotes subfasciatus; (e) from the order of the dipterans (Diptera), for example, Anastrepha ludens, Ceratitis capitata, Contarinia sorghicola, Dacus cucurbitae, Dacus oleae, Dasineura brassicae, Delia coarctata, Delia radicum, Hydrellia griseola, Hylemyia platura, Liriomyza sativae, Liriomyza trifolii, Mayetiola destructor, Orseolia oryzae, Oscinellafrit, Pegomya hyoscyami, Phorbia antiqua, Phorbia brassicae, Phorbia coarctata, Rhagoletis cerasi, Rhagoletis pomonella, (f) furthermore Aedes aegypti, Aedes vexans, Anopheles maculipennis, Chrysomya bezziana, Chrysomya hominivorax, Chrysomya macellaria, Cordylobia anthropophaga, Culex pipielis, Fannia canicularis, Gasterophilus intestinalis, Glossina morsitans, Haematobia irritans, Haplodiplosis equestris, Hypoderma lineata, Lucilia caprina, Lucilia cuprina, Lucilia sericata, Musca domestica, Muscina stabulans, Oestrus ovis, Tabanus bovinus, Simulium damnosum; (g) from the order of the thrips (Thysanoptera), for example, Frankliniella fusca, Frankliniella occidentalis, Frankliniella tritici, Haplothrips tritici, Heliothrips haemorrhoidalis, Scirtothrips citri, Thrips oryzae, Thripspalmi, Thrips tabaci; (h) from the order of the hymenopterans (Hymenoptera), for example, Athalia rosae, Atta cephalotes, Atta sexdens, Atta texana, Hoplocampa minuta, Hoplocampa testudinea, Iridomyrmes humilis, Iridomyrmex purpureus, Monomorium pharaonis, Solenopsis geminata, Solenopsis invicta, Solenopsis richteri, Technomyrmex albipes; (i) from the order of the heteropterans (Heteroptera), for example, Acrosternum hilare, Blissus leucopterus, Cyrtopeltis notatus, Dysdercus cingulatus, Dysdercus intermedius, Eurygaster integriceps, Euschistus impictiventris, Leptoglossus phyllopus, Lygus hesperus, Lygus lineolaris, Lygus pratensis, Nezara viridula, Piesma quadrata, Solubea insularis, Thyanta perditor; (j) from the order of the homopterans (Homoptera), for example, Acyrthosiphon onobrychis, Acyrthosiphon pisum, Adelges laricis, Aonidiella aurantii, Aphidula nasturtii, Aphis fabae, Aphis gossypii, Aphis pomi, Aulacorthum solani, Bemisia tabaci, Brachycaudus cardui, Brevicoryne brassicae, Dalbulus maidis, Dreyfusia nordmannianae, Dreyffusia piceae, Dysaphis radicola, Empoasca fabae, Eriosonma lanigerum, Laodelphax striatella, Macrosiphum avenae, Macrosiphum euphorbiae, Macrosiphon rosae, Megoura viciae, Metopolophium dirhodum, Myzus persicae, Myzus cerasi, Nephotettix cincticeps, Nilaparvata lugens, Perkinsiella saccharicida, Phorodon humuli, Psylla mali, Psylla piri, Psylla pyricola, Rhopalosiphum maidis, Schizaphis graminum, Sitobion avenae, Sogatella furcifera, Toxoptera citricida, Trialeurodes abutilonea, Trialeurodes vaporariorum, Viteus vitifolii; k) from the order of the termites (Isoptera), for example, Calotermes flavicollis, Coptotermes spp, Leucotermes flavipes, Macrotermes subhyalinus, Nasutitermes spp such as Nasutitermes walkeri, Odontotermes formosanus, Reticulitermes lucifugus, Termes natalensis; (l) from the order of the orthopterans (Orthoptera), for example, Gryllotalpa gryllotalpa, Locusta migratoria, Melanoplus bivittatus, Melanoplus femur-rubrum, Melanoplus mexicanus, Melanoplus sanguinipes, Melanoplus spretus, Nomadacris septemfasciata, Schistocerca americana, Schistocerca peregrina, Stauronotus maroccanus, Schistocerca gregaria, (m) furthermore Acheta domestica, Blatta orientalis, Blattella germanica, Periplaneta americana; (n) from the order of the phthirapterans (Phthiraptera), for example, Mallophaga, such as Damalina spp., and Anoplura such as Linognathus and Haematopinus spp.\",\n \"; (O) from the order of the hemipterans (Hemiptera), for example, Aphis, Bemisia, Phorodon, Aeneolamia, Empoasca, Parkinsiella, Pyrilla, Aonidiella, Coccus, Pseudococcus, Helopeltis, Lygus, Dysdercus, Oxycarenus, Nezara, Aleyrodes, Triatoma, Psylla, Myzus, Megoura, Phylloxera, Adelges, Nilaparvata, Nephotettix or Cimwx spp.\",\n \"; (p) from the order of the siphonapterans (Siphonaptera), for example, Ctenocephalides or Pulex spp.\",\n \"; (q) from the order of the thysanurans (Thysanura), for example, Lepisma spp.\",\n \"; (r) from the order of the dermapterans Dermaptera), for example, Forficula spp.\",\n \"; and (s) from the order of the psocopterans (Psocoptera), for example, Peripsocus spp.\",\n \"Arachnids contemplated by the present invention include, but are not limited to, spiders and scorpions and especially mites such as phytophagous mites (Acari), such as Aculops lycopersicae, Aculops pelekassi, Aculus schlechtendali, Brevipalpus phoenicis, Bryobia praetiosa, Eotetranychus carpini, Eutetranychus banksii, Eriophyes sheldoni, Oligoinychus pratensis, Panonychus ulmi, Panonychus citri, Phyllocoptruta oleivora, Polyphagotarsonenmus latus, Tarsonemuspallidus, Tetranychus cinnabarinus, Tetranychus kanzawai, Tetranchus pacificus, Tetranychus urticae, ticks, such as Amblyomma americanum, Amblyomma variegatum, Argas persicus, Boophilus annulatus, Boophilus decoloratus, Boophilus microplus, Dermacentor silvarum, Hyalomma truncatum, Ixodes ricinus, Ixodes rubicundus, Ornithodorus moubata, Otobius megnini, Rhipicephalus appendiculatus and Rhipicephalus evertsi, and animal-parasitic mites, such as Dermanyssus gallinae, Psoroptes ovis and Sarcoptes scabiei.\",\n \"Helminths falling within the scope of the present invention may be selected from cestodes such as fish tapeworm, pork tapeworm, beef tapeworm, and dwarf tapeworm; trematodes such as from the genera Metagonimus and Heterophyes; and nematodes such as but not limited to filariid, ascarid, strongyle and trichostrongyle nematodes of the genera Acanthocheilonema, Aelurostrongylus, Ancylostoma, Angiostrongylus, Ascaris, Brugia, Bunostomum, Dictyocaulus, Dioctophyme, Dipetalonema, Dirofilaria, Dracunculus, Filaroides, Lagochilascaris, Loa, Mansonella, Muellerius, Necator, Onchocerca, Parafilaria, Parascaris, Protostrongylus, Setaria, Stephanofilaria, Strongyloides, Strongylus, Thelazia, Toxascaris, Toxocara, Trichinella, Uncinaria and Wuchereria.\",\n \"Suitable molluscs include those of the Gastropoda class examples of which include snails, slugs, conchs, and whelks.\",\n \"Protozoa may be selected for example from Plasmodia, Toxoplasma, Leishmania, Trypanosoma, Giardia, Entamoeba, Acanthamoeba, Nagleria, Hartmanella, Balantidium, Babesia, Cryptosporidium, Isospora, Microsporidium, Trichomonas or Pneumocystis species.\",\n \"Viruses may be selected from RNA viruses or DNA viruses, which include but are not limited to Human Immunodeficiency Virus (HIV), Poliovirus, Influenza virus, Rous Sarcoma virus, Flaviviruses such as Japanese encephalitis, Influenza virus, Respiratory Syncytial Virus, Hepatitis virus, Parvovirus, Rotavirus, Coronavirus, Adenovirus and Herpesviruses such as Papillomavirus and Epstein-Barr virus.\",\n \"The present invention also extends to methods for producing resistance in plants to pests including, but not limited to, insects, arachnids, helminths, molluscs, protozoa and viruses by crossing a plant expressing a β-dione compound according to the invention with pest susceptible lines.\",\n \"Crossing a β-dione-producing plant into a pest susceptible background would produce a resistant plant with a high level of pest resistance.\",\n \"Plants that could be made pest resistant include, but are not limited to, dicotyledonous plants, especially trees and more especially members of the Myrtaceae family.\",\n \"For example E. cloeziana commonly known as Gympie Messmate is one of the many Eucalyptus species grown for hard wood production.\",\n \"However the oil present in this chemotype does not contain β-diones and hence an intra species cross with the unique North Queensland tasmanone chemotype would introduce this phenotypic trait.\",\n \"Such a process would be readily applicable to other Eucalyptus species of commercial interest.\",\n \"Interspecific crossing within the Myrtaceae family is well established to those skilled in the art and inclusion of β-dione as an additional trait into formal breeding programs is acknowledged.\",\n \"Suitable β-dione-producing plants may be selected from the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.\",\n \"Preferably, the volatile oil-bearing plant is selected from genera of the Myrtaceae family including, but not limited to, Angophora, Austromyrtus, Backhousia, Baeckea, Callistemon, Corymbia, Darwinia, Eucalyptus, Kunzea, Leptospermum, Melaleuca, Syzygium and Xanthostemon.\",\n \"Preferred 16-dione-producing plants are Leptospermum mornsonii, Eucalyptus bensonii, Eucalyptus megacornuta, Eucalyptus pilularis, Eucalyptus cornuta, Eucalyptus baxteri, Eucalyptus macrorhyncha, Eucalyptus cloeziana, Melaleuca cajuputi, Eucalyptus jensenii, Backhousia angustifolia and Leptospennum scoparium.\",\n \"A particularly preferred β-dione-producing plant is Eucalyptus cloeziana.\",\n \"As used herein, the term “plant” includes reference to whole plants, plant organs (e.g., leaves, stems, roots, etc.\",\n \"), seeds and plant cells and progeny of same.\",\n \"Plant cell, as used herein includes, without limitation, seeds suspension cultures, embryos, meristematic regions, callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen, and microspores.\",\n \"The class of plants which can be used in the methods of the invention is generally as broad as the class of higher plants amenable to transformation techniques, including both monocotyledonous and dicotyledonous plants.\",\n \"Thus, the present invention contemplates conventional plant breeding methods to transfer the genetic material associated with β-dione production via crossing and backcrossing.\",\n \"Such methods will comprise the steps of: (1) sexually crossing the β-dione-producing plant with a plant from a pest susceptible taxon; (2) recovering reproductive material from the progeny of the cross; and (3) growing β-dione-producing/pest-resistant plants from the reproductive material.\",\n \"Where desirable or necessary, the agronomic characteristics of the susceptible taxon can be substantially preserved by expanding this method to include the further steps of repetitively: (1) backcrossing the pest-resistant progeny with pest-susceptible plants from the susceptible taxon; and (2) selecting for expression of a β-dione (or an associated marker gene) among the progeny of the backcross, until the desired percentage of the characteristics of the susceptible taxon are present in the progeny along with the gene or genes imparting β-dione activity.\",\n \"By the term “taxon” herein is meant a unit of botanical classification.\",\n \"It thus includes, genus, species, cultivars, varieties, variants and other minor taxonomic groups which lack a consistent nomenclature.\",\n \"In order that the invention may be readily understood and put into practical effect, particular preferred embodiments will now be described by way of the following non-limiting examples.\",\n \"EXAMPLES Example 1 β-Triketone-Containing Oils Obtained from Australian Myrtaceae Species Australia has an extensive number of volatile oils from species of the Myrtaceae that are rich in a diversity of structurally related constituents known as β-triketones.\",\n \"These oils often show not only a high yield of oil, but also a high degree of biosynthetic selectivity that produces β-triketones in a high proportion of the oil composition.\",\n \"The major constituents of the published Myrtaceae essential oils (Hellyer, 1968; Boland and Brophy, 1990, 1993; Brophy, et al., 1995; Bignall et al., 1997; Southwell and Brophy 2000) are listed in Table 1 and their structures are included in FIG.\",\n \"1.TABLE 1 β-TRIKETONE PROFILES OF AUSTRALIAN ESSENTIAL OILS Species Yield β-Triketone (%) Distribn Backhousia angustifolia 2.5 Angustione (85) QLD Backhousia angustifolia 2.5 Dehyroangustione (80) QLD Eucalyptus cloeziana 3.0 Tasmanone (95) QLD Eucalyptus suberea 1.4 Tasmanone (94) WA Eucalyptus lateritica 0.9 Tasmanone (37), WA lateriticone (14) Eucalyptus camfieldii Tasmanone (40) NSW Leptospermum scoparium 0.4 Leptospermone (19), NSW, NZ Flavesone (8), Isoleptospermone (5) Eucalyptus grandis 0.6 Leptospermone (20), NSW Flavesone (13), Isoleptospermone (3) Eucalyptus agglomerata Agglomerone (40) NSW Eucalyptus mckieana Agglomerone (60) NSW Eucalyptus bensonii 2.5 Agglomerone (72) Eucalyptus insularis 1 Agglomerone (19) WA Eucalyptus jensenii 0.3 Jensenone (70) NT Eucalyptus papuana 0.7 Papuanone (40) Nth Aus Leptospermum morrisonii 1.8 Grandiflorone (58) NSW Melaleuca cajeputi Platyphyllol Nth Aus Xanthostemon Xanthostemone oppositifolius The β-triketones obtained from selected Myrtaceous volatile oils were shown to have significant insecticidal and/or acaricidal activity.\",\n \"Example 2 Insecticidal Activity Initial insecticidal screening against two important arthropod species, two-spotted mite (Tetranychus urticae) and diamond back moth (Plutella xylostella) lavae highlighted three oils on the basis of efficacy, oil yield, oil profile and ease of recollection (Table 2).\",\n \"Where feasible, the LD50 and LD95 values were determined.\",\n \"TABLE 2 PERCENTAGE MORTALITY OF THREE EFFICACIOUS OILS Diamond Two Spotted Mite Back Moth % Mortality % Mortality Species (0.5%/1.0%) (0.5%/1.0%) Backhousia angustifolia 98/98 100/100 Backhousia angustifolia 100/100 100/100 Eucalyptus cloeziana 100/100 70/100 Insecticidal tests using the oil from fresh plant recollections and steam distillations varied occasionally and, in this regard, it is believed that improving storage conditions including temperature, light and exposure to air and inclusion of a dessicant can enhance the stability of the active fraction of such oils.\",\n \"E. cloeziana oil continued to show high potency against both insect tests.\",\n \"This oil exhibited an LD95 between 0.04-0.20% (depending on formulation and treatment) against two-spotted mite.\",\n \"An LD95 of 0.10% was observed against 1st instar lavae of diamond back moth and this rose to 0.78% when tested against 3rd instar lavae.\",\n \"In additional preliminary investigations with greenhouse thrips (Heliothrips haemorrhoidalis), a 0.1% concentration of E. cloeziana oil induced 100% mortality.\",\n \"Additional work was carried out on the E. cloeziana oil to explore the contribution the various components make to the overall efficacy of this oil.\",\n \"Fractions 1, 3, 4, and 5 outlined in Example 3 were screened against two-spotted mite.\",\n \"Fractions 1 and 5 showed no significant insecticidal effect.\",\n \"Fractions 3 and 4, comprising 98% and 99% tasmanone were active and showed little difference to the activity of the whole oil.\",\n \"This suggests that not only is tasmanone the principle component in the oil, but it is also the principle bioactive constituent.\",\n \"It is also reasonable to assume that as the activity of E. cloeziana oil has been demonstrated against a number of different arthropod species, namely a mite (T. urticae), a caterpillar (P. xylostella) and a thrips (H. haemorrhoidalis), its insecticidal activity is broad in nature.\",\n \"Example 3 Chemistry Chemical analysis (GC-MS) of the steam-distilled oils from the collected plants in this work are summarised in Table 3.TABLE 3 β-TRIKETONE PROFILES OF SELECTED OILS Plant Source Principle Component* (%) Backhousia angustifolia-I Dehydroangustione 85% Backhousia angustifolia-II Dehydroangustinone (80%) Backhousia angustifolia-III Angustinone (65%) Backhousia angustifolia-IV Angustinone (28%) Eucalyptus cloeziana Tasmanone (84-96%) Melaleuca cajuputi subsp platyphylla Platyphyllol (64-71%) *As determined by gas chromatography The most promising oils were derived from B. angustifolia, M. cajuputi subsp platyphylla and E. cloeziana and these were then subjected to additional chemical fractionation.\",\n \"The lower insecticidal activity observed in the recollected B. angustifolia (I) was in part attributed to the lower levels of β-triketone.\",\n \"The level of β-triketone was elevated by the removal of the more volatile monoterpenes using vacuum distillation.\",\n \"One of the most efficacious oils was from E. cloeziana, a tasmanone (84-96%) rich oil with additional terpenes and β-triketones (FIG.\",\n \"2, Table 4), which displayed consistent activity at every stage of processing and formulation.\",\n \"This oil was, therefore, fractionated using column chromatography on silica gel with a hexane-diethyl ether gradient and a final methanol elution.\",\n \"The profiles of the fractions are summarised in Table 5.TABLE 4 CHEMICAL PROFILE OF E. cloeziana OIL Peak No Compound Composition (%) 1 a-pinene 0.5-1.9 2 B-pinene 1.5-5.7 3 Limonene 0.1-0.6 4 a-terpineol 0.7-2.0 5 Globulol 0.01-0.5 6 Agglomerone 0.01-0.6 7 Tasmanone 84-96 8 Lateriticone 0.2-0.7 9 Isolateriticone 0.3-1.2 TABLE 5 FRACTIONS CUT FROM E. cloeziana OIL Fraction No Solvent System Composition Amount F1 Hexane Hydrocarbons 73 mg F2 Hex:Et2O (9:1) 80% Tasmanone 4 mg F3 Hex:Et2O (1:1) 98% Tasmanone 3.66 g F4 Hex:Et2O (9:1) +99% Tasmanone 694 mg F5 MeOH Terpene alcohols 64 mg 1H NMR data (FIG.\",\n \"3) were recorded on F4 and confirmed the structure of tasmanone.\",\n \"Moreover the compound exists in solution (CDCl3) in tautomeric forms (FIG.\",\n \"4) in the ratio 2:0:1 (A:B:C).\",\n \"Another efficacious oils was from M. cajuputi subsp platyphylla, a platyphyllol (64-71%) rich oil (Table 6), which also displayed consistent activity at every stage of processing and formulation.\",\n \"TABLE 6 TYPICAL CHEMICAL PROFILE OF M. cajuputi subsp platyphylla OIL Peak No Compound Composition (%) 1 a-pinene Tr-0.8 2 1,8-cineole Tr-0.7 3 B-caryophyllene 0.6-3.2 4 Humulene 0.6-1.3 5 Spathulenol 4.0-9.0 6 Caryophyllene oxide Tr-3.6 6 Platyphyllol 64-71 7 MW 234 - unknown 4.3 *As determined by gas chromatography Example 4 Phytotoxicity Initial investigations using leaves and leaf discs of several plant species including French bean (Phaseolus vulgaris) lemon (Citrus limon) Orange (Citrus sinensis) and Cabbage (Brassica oleracea) indicated that phytotoxicity did not occur for most oils below 0.5%.\",\n \"More detailed investigations using soft intact leaves of young greenhouse-grown French bean subsequently showed E. cloeziana oil applied as a spray caused some phytotoxicity at concentrations of 0.5% and above.\",\n \"Example 5 Toxicity of E. Cloeziana Oil Bacterial Reverse Mutation Assay This study investigated the potential of E. cloeziana oil to induce reverse mutations at the histidine locus in the genome of one strain of Salmonella typhimurium TA100 in the presence and absence of a metabolic activation system (mammalian microsomal enzymes, S9 mix).\",\n \"The test sample was dissolved in dimethyl sulfoxide (DMSO).\",\n \"In this assay, an E. cloeziana oil test sample did not induce an appropriate-fold increase (a 2-fold increase for TA100) in the mean revertants per plate in the tester strain TA100 over the mean revertants per plate of the appropriate vehicle control.\",\n \"Accordingly, the test sample was considered to be non-mutagenic under the conditions of the assay.\",\n \"Acute Oral Sighting The acute oral toxicity of E. cloeziana oil was investigated in ten (10) Sprague Dawley Specific Pathogen Free female rats (groups of 2) at doses of 500, 250, 125 and 50 mg/kg.\",\n \"The experimental procedure was based on OECD guidelines for the testing of chemicals, No.\",\n \"420.Clinical signs of toxicity occurred between one (1) and twenty-four (24) hours after dosing.\",\n \"Both animals in the 500 mg/kg group exhibited subdued behaviour, partial eye closure, slow breathing, reduced motor activity, ataxia followed by death within 24 hours.\",\n \"The animals in the 250 mg/kg group exhibited subdued behaviour, partial eye closure, slow breathing, social isolation, and reduced motor activity, and had returned to normal by 24 hours after dosing.\",\n \"The animals in the 125 mg/kg group exhibited subdued behaviour, partial eye closure, slow breathing, piloerection and reduced motor activity, and had returned to normal by 24 hours after dosing.\",\n \"The animals in the 50 mg/kg group did not show any signs of toxicity during the seven day experimental period.\",\n \"There were no other clinical abnormalities in any animal throughout the seven (7) day observation period.\",\n \"The stomach of one animal in the 250 mg/kg group (5F) had a single ulcer.\",\n \"There were no other gross abnormalities in the major organs of any animal at autopsy.\",\n \"Based on the results obtained from this study 50, 100 and 200 mg/kg can be selected for a main study as the maximum non-toxic dose, intermediate dose and high dose, respectively.\",\n \"Full Study The acute oral toxicity of E. cloeziana oil was investigated in thirty (30) Sprague Dawley Specific Pathogen Fee rats (15 males and 15 females) at doses of 50, 100 and 200 mg/kg.\",\n \"These doses were chosen following a dose range finding in the above study.\",\n \"The experimental procedure was based on OECD guidelines for the testing of chemicals, No.\",\n \"420.The acute NOAEL of E. cloeziana oil was determined to be 50 mg/kg, and the MTD was 200 mg/kg under the conditions of this study.\",\n \"Acute Dermal The acute dermal toxicity of E. cloeziana oil was investigated in ten (10) Sprague Dawley Specific Pathogen Free rats (5 males and 5 females) at a dose of 2000 mg/kg.\",\n \"A preliminary study (SIGHTING) indicated no signs of toxicity at this dose.\",\n \"The experimental procedure was based on OECD guidelines for the testing of chemicals, No.\",\n \"420.No clinical abnormalities, skin irritations or body weight losses were observed in any animal throughout the fourteen (14) day observation period.\",\n \"No deaths occurred.\",\n \"No abnormalities were seen in the major organs at necropsy.\",\n \"The rat acute dermal LD50 of E. cloeziana oil was determined to be greater than 2000 mg/kg under the conditions of this study.\",\n \"Skin Irritancy/Corrosion The potential of a test sample of E. cloeziana oil to provoke skin irritation/corrosion reactions was investigated using a primary dermal irritation/corrosion test in three (3) New Zealand White albino rabbits (OECD Guidelines for the Testing of Chemicals, No.\",\n \"404).\",\n \"The results obtained from this study indicated that E. cloeziana oil is a non-irritant according to the National Occupational Health and Safety Commission (NOHSC) “Approved Criteria for Classifying Hazardous Substances [NOHSC:1008(1999)]”.\",\n \"Example 6 Isolation and Purification of β-Triketones from Australian Myrtaceae Species Plant collections were commissioned at the Mt.\",\n \"Annan Botanic Garden, Sydney, Australia, and the Darwin Botanic Garden, Northern Territory, Australia to provide potential sources of a range of β-triketone isolation.\",\n \"The plants investigated for β-triketone exploration are listed in Table 6.TABLE 6 PLANTS SOURCED FOR β-TRIKETONE EXPLORATION β-triketone # Source Plant Plant Location Accession # anticipated 1 Leptospermum morrisonii Mt.\",\n \"Annan, NSW 873247 grandiflorone 2 Eucalyptus bensonii Mt.\",\n \"Annan, NSW 881045 agglomerone 3 Eucalyptus megacornuta Mt.\",\n \"Annan, NSW 831078 jensenone+ 4 Eucalyptus pilularis Mt.\",\n \"Annan, NSW 873166 torquatone 5 Eucalyptus cornuta Mt.\",\n \"Annan, NSW 852618 jensenone+ 6 Eucalyptus baxteri Mt.\",\n \"Annan, NSW 860999 agglomerone/ tasmanone 7 Eucalyptus macrorhyncha Mt.\",\n \"Annan, NSW 860896 conglomerone 8 Eucalyptus cloeziana Lappa, QLD PF 2513 tasmanone 9 Melaleuca cajuputi ssp Bensbach, WP, PNG KW16-19 platyphyllol platyphyla 10 Eucalyptus jensenii NT Botanic Gardens, RK114- jensenone Darwin, NT LUS 11 Backhousia angustifolia Wilgavale, Texas, PF1712 dehydroangustione QLD 12 Backhousia angustifolia Didcott Creek, QLD PF 1708 angustione 13 Leptospermum scoparium NA commercial oil NA flavesone, sample isoleptospermone, leptospermone 14 Eucalyptus conjuncta Mt Annan 854097 conglomerone +Oil contains other β-triketone constituents All plants, apart from 9 and 13, were steam distilled to yield various quantities of essential oil.\",\n \"All oils were analysed by Gas Chromatography Mass Spectrometry (GCMS) to determine the presence and abundance of β-triketones.\",\n \"Based on this information, particular oils were targeted for isolation and purification of β-triketones using wet chemistry and preparative HPLC techniques.\",\n \"The β-triketones listed in Table 7 were isolated in quantities adequate for insecticidal screening.\",\n \"A minimum of 50 mg of each compound was required.\",\n \"Isoleptospermone and leptospermone were difficult to separate due to their structural similarity as were angustione and dehydroangustione.\",\n \"Consequently, mixtures of these compounds, where one isomer was significantly more abundant, were provided for insecticidal screening as this will still allow for differentiation in observed activity.\",\n \"TABLE 7 β-TRIKETONES ISOLATED FOR INSECTICIDAL SCREENING Amount sent Sample Purity % by for screening β-triketones Plant source GCMS (mg) grandiflorone Leptospermum morrisonii 100.00 99.5 jensenone Eucalyptus jensenii 100.00 67.9 dehydroangustione Backhousia angustifolia 95.02 165.02 PF1712 angustione Backhousia angustifolia 66.30 (33.70% 107.1 PF1708 dehydroangustione) agglomerone Eucalyptus bensonii 99.28 109.2 flavesone Leptospermum scoparium 99.32 101.8 isoleptospermone Leptospermum scoparium 33.10 (66.90% 113.4 leptospermone) leptospermone Leptospermum scoparium 95.33 77.0 tasmanone Eucalyptus cloeziana 99.92 110.1 platyphyllol Melaleuca cajuputi subsp.\",\n \"99.62 255.6 platyphylla The chemical structures and identities of the β-triketones isolated were confirmed by GCMS and Nuclear Magnetic Resonance (NMR) analysis.\",\n \"Example 7 Efficacy of E. cloeziana Oil on Target Organisms Target Organisms Two spotted mite (TSM) Tetranychus urticae Koch [Acarina: Tetranychidae] were collected from a mass culture maintained at the University of Western Sydney's Hawkesbury Campus in Richmond, NSW, Australia They were reared on potted French beans (Phaseolus vulgaris L [Fabales: Fabaceae] in a glasshouse maintained at 25±5° C., 65±5% RH and 14 h D:L photoperiod.\",\n \"Only young females were selected for bioassay.\",\n \"Adult parthenogenetic female greenhouse thrips (GHT), Heliothrips haemorrhoidalis Bouché (Thysanoptera: Thripidae) of similar age were obtained from a colony reared on orange fruits and maintained in an insectary at UWS Hawkesbury under conditions of 25±3° C., 65% RH and 16 h D:L photoperiod Young nymphs of citrus aphids Toxoptera citricida (Kirkaldy) (Hemiptera: Aphididae) were collected from lemon seedlings grown under glasshouse conditions at UWS Hawkesbury.\",\n \"Workers of the termite Nasutitermes walkeri Hill (Isoptera: Termitidae) were collected from a laboratory culture at UWS Richmond which was initiated from soil, termitaria and wood on which termites were feeding were field collected at Richmond NSW, and maintained in a darkened container under conditions of 25±2° C., 35-68% RH.\",\n \"Termites were fed on wood collected from near the original nest.\",\n \"Moistened soil from the nest together with paper towel were placed on top of the nest and made it possible to maintain this culture for several months in the laboratory at UWS Hawkesbury.\",\n \"Workers of the whitefooted house ant Technomyrmex albipes (F. Smith) (Hymenoptera: Formicidae) were field collected at UWS Hawkesbury by baiting in an empty glass jar containing sugar granules.\",\n \"Pupae of housefly, Musca domestica L (Diptera: Muscidae) and different stages of American Cockroach, Periplaneta americana L (Blattodea: Blattidae) were initially supplied by C.E.R.I.T and maintained in laboratory culture at UWS Hawkesbury.\",\n \"Tomato russet mites (TRM) Aculops lycopersici (Massee) (Acarina: Eriophyidae) were collected from infested tomato plants near Riverstone, NSW.\",\n \"Mixed sex adults of the mosquito Culex quinquefaciatus (Diptera: Culicidae) were supplied by C.E.R.I.T, held at the in the Centre for Horticulture & Plant Science, University of Western Sydney, Richmond NSW and treated one day after arrival.\",\n \"Workers of the honeybee Apis mellifera (Hymenoptera: Apidae) were collected from several field hives maintained at the apiary in the Centre for Horticulture & Plant Sciences, University of Western Sydney, Richmond NSW Adults of ash-white-flies Aleaurocanthus woglumi (Homoptera: Aleyrodidae) were field collected from an ornamental pear (Prunus sp) in the Centre for Horticulture & Plant Sciences, University of Western Sydney, Richmond NSW.\",\n \"Drug store beetle Sitodrepa panicea (Coleoptera: Anobiidae) were reared on curry powder under laboratory conditions of 25±1° C. and 65±5% RH.\",\n \"Mixed age groups of snails Helix apersa (Mollusca: Gastropoda) were collected from infested plants in the Centre for Horticulture & Plant Science, University of Western Sydney, Richmond NSW.\",\n \"Bioassays TSM From an E. cloeziana oil extract (containing 85% tasmanone), 1.1765 g was dissolved in 5 mL ethyl alcohol and distilled water containing 200 ppm Triton X-100™ was added to prepare a 1% stock solution.\",\n \"From this homogenised stock solution, further serial dilutions of 0.0125, 0.025, 0.05, 0.10, and 0.12% were prepared by mixing the required amount of stock solution in distilled water and Triton X-100 solution.\",\n \"Each treatment was conducted on 60-80 TSM, which were evenly distributed on four French bean leaf discs (25 mm diam) contained in 90 mm diam.\",\n \"petri dishes.\",\n \"The leaf discs were placed with their underside uppermost on moist absorbent cotton wool covered with muslin netting.\",\n \"Water was added to the dishes daily to prevent desiccation of the leaf discs.\",\n \"Five mL aliquots (unless otherwise stated) were applied to each petri dish with a Potter precision spray tower as described by Herron et al (1995).\",\n \"The average mass of solution applied to each dish was calculated to be 3.95 mg/cm2.Mortality was recorded 24 h after treatment.\",\n \"Death was recognised by the absence of movement when the test organisms were mechanically stimulated.\",\n \"Data were analysed using SPSS for Windows™ Version 7.Probit analysis was carried out for dose-mortality data and heterogeneity of regressions was determined by the Pearson chi-squared characteristic.\",\n \"In addition, TSM was treated with E. cloeziana oil extract in combination with paraffin oil.\",\n \"In particular, E. cloeziana oil extract at levels of 0.1, 0.2, 0.3, 0.4 and 0.5 g were weighed out and each was made up to a weight of 10.0 g with a formulated paraffin oil (BioPest®), which was then sonicated for 10 min.\",\n \"A 1.0% v/v of each blend was prepared by mixing 1.0 mL with distilled water in a 100.0 mL volumetric flask.\",\n \"TSM were transferred to the petri dish following the same standard method Five-mL aliquots were applied to each petri dish and mortality was recorded 24 h after treatment.\",\n \"All blends of E. cloeziana oil extract with paraffin oil produced 100.0% mortality, compared with Biopest® alone which caused only 35.5±6.9 mortality.\",\n \"The lowest concentration of E. cloeziana oil extract tested in this combination (ie.\",\n \"0.01%) resulted in 100% mortality in TSM, which is significantly lower than that reported earlier in this document to produce 100% mortality with E. cloeziana oil extract alone (>0.06%).\",\n \"Greenhouse Thrips The same experimental procedure used for TSM was repeated for GHT except that lemon leaf discs were used instead of French beans.\",\n \"The required number of adult thrips for each treatment was transferred with a fine brush to the underside of a lemon disc of the same diameter (6 cm) as the base of a petri dish.\",\n \"The lemon disc was mounted on agar with its adaxial side uppermost.\",\n \"Immediately after treatment the petri dish was covered with perforated plastic wrap.\",\n \"Mortality was assessed 24 h after treatment.\",\n \"Tomato Russet Mite The same experimental procedure for TSM was repeated except that tomato leaf discs were used instead of French bean.\",\n \"Brown Citrus Aphid Lemon leaf discs 2.5 cm diam.\",\n \"were cut from tender young leaves and mounted on moistened absorbent cotton wool in 90 mm petri dish with their adaxial surface uppermost.\",\n \"Each petri dish contained four leaf discs.\",\n \"Uniform early instar nymphs were then transferred with a fine brush to the leaf discs (each containing 8-10 nymphs).\",\n \"A Potter tower was used to apply 5 mL aliquots to each petri dish.\",\n \"A control (solvent and surfactant only) was also included in the assessment.\",\n \"Mortality was assessed 24 h after treatment.\",\n \"Termites Twenty uniform termite workers were transferred to 90 mm petri dishes lined with the same diameter moistened filter paper (Whatman No 2).\",\n \"A preliminary trial was carried out using a Potter tower to apply 5 mL aliquots of each concentration.\",\n \"Using this method, all termite workers died 4 h after treatment in all concentrations, including the lowest concentration of 0.015%.\",\n \"There was no mortality recorded in the blank control treatment, and all workers remained alive and active for >48 h after treatment.\",\n \"Identical results were obtained from these investigations, whether the petri dishes were covered or uncovered after application of the E. cloeziana oil extract.\",\n \"Subsequent investigations further assessed efficacy of E. cloeziana oil extract by releasing termite workers on fresh dried residues.\",\n \"This was carried out by uniformly distributing one mL of each concentration over the entire surface area of a 90 mm diam filter paper.\",\n \"When the paper was air dry, 20 termite workers were placed in each petri dish.\",\n \"One hundred percent mortality was recorded in all four replicates even at the lowest concentration applied (0.015% w/v=150 ppm).\",\n \"This suggests that the plant extract is a highly toxic contact poison to termites.\",\n \"Ants The same experimental procedure for termites was repeated for ants, with the required number of worker ants for each treatment transferred with a fine brush to the filter paper containing a fresh dried residue of the E. cloeziana oil extract.\",\n \"Immediately after release of the ants the petri dish was covered with perforated plastic wrap, which enabled any excess vapours to escape while retaining the ants.\",\n \"Mortality was assessed 4 h after treatment.\",\n \"In all 4 replicates 100% mortality was obtained at concentrations as low as 0.0075% w/v ai when applied at a rate of 1.0 mL to a 90 mm diam.\",\n \"filter paper.\",\n \"Houseflies One percent E. cloeziana oil extract was prepared using pure acetone as a diluent.\",\n \"From this solution, further serial dilutions were prepared by adding the required amount in acetone.\",\n \"Five mL aliquots of each concentration of E. cloeziana oil extract were dispensed into 500 mL kilner jars.\",\n \"The kilner jars were immediately rotated until dryness to coat the inner surface uniformly with the E. cloeziana oil residue.\",\n \"After complete dryness, 30-50 pupae were transferred to a series of clean uncovered petri dishes(45 mm diam.\",\n \"), one of which was placed inside each jar.\",\n \"The jar mouth was then covered with nylon netting supported by a rubber band.\",\n \"All adult houseflies started to emerge from pupae after 48 h and most emerged within a 3 h period.\",\n \"Flies were fed 5% sugar solution soaked in absorbent cotton wool.\",\n \"Jars were maintained in an incubator at 29° C. Mortality was assessed at the end of the third day (i.e., approx.\",\n \"72 h) after application of the E. cloeziana oil residues in the kilner jar and the placement of pupae inside the jars.\",\n \"(This comprised 48 h for pupae to emerge and 24 h exposure to E. cloeziana oil residues which were now 48 h old).\",\n \"Flies were observed to die within a few hours after emergence, whereas in the control they remained alive for >48 h after emergence.\",\n \"The total number of adult houseflies that emerged in each kilner jar was counted and their mortality was recorded.\",\n \"American Cockroaches Tests were conducted on 10-20 three months old nymphs (mean individual mass 0.2-0.3 g) and replicated three times.\",\n \"One mL of 1.0% E. cloeziana oil extract in acetone was uniformly distributed on 90 mm diam.\",\n \"Whatman No 2 filter paper.\",\n \"A control treatment was also carried out using 1 mL acetone only (ie., minus E. cloeziana oil extract).\",\n \"After complete dryness of the filter paper, the required number of cockroaches was transferred inside the kilner jars, and were fed dry dog food.\",\n \"Kilner jar necks were covered with muslin netting supported by rubber bands.\",\n \"Mortality was assessed 24 h after releasing the cockroaches, and death was recognised by the absence of movement when the test animals were mechanically stimulated.\",\n \"Adult Mosquitoes A 0.3576 g of 85% ai of E. cloeziana oil extract was dissolve in pure acetone as a diluent to give approximately 0.304% concentration stock solution.\",\n \"From this solution, further serial dilutions 0.152, 0.076, 0.0043 and 0.00215 were prepared by adding the required amount in acetone.\",\n \"Aliquots (2.5 mL) of each concentration of E. cloeziana oil extract were dispensed into 500 mL kilner jars with total internal surface area as 286.53 cm2.The kilner jars were immediately rotated to coat the inner surface uniformly with the E. cloeziana oil extract residue, until dry.\",\n \"Once completely dry, 10-25 mixed sex adult mosquitos were released into each kilner jar by allowing the mosquitoes to fly from a darkened cage into the naturally lit jars.\",\n \"The jar mouth was subsequently placed on 110 mm diam.\",\n \"filter paper onto which had been placed a yellow sponge soaked in 7.0% sugar solution.\",\n \"Treated jars were kept under laboratory temperature and humidity conditions, (viz.\",\n \"24±1° C. and 65±5% RH respectively).\",\n \"Mortality was assessed 24 h after releasing the mosquitoes in the jars.\",\n \"Honey Bees Ten worker honey bees, anaesthetised with carbon dioxide, were transferred to 90 mm diam.\",\n \"petri dishes lined with moistened filter paper.\",\n \"Five mL aliquots were applied using a Potter Spray Tower while the bees were still anaesthetised.\",\n \"Yellow sponges soaked in 7% sugar solution were then placed inside petri dishes for bee sustenance, and the bees after treatment were placed in these dishes.\",\n \"The lids that were perforated and covered with muslin netting were placed on the dishes, and mortality was assessed 24 hours after treatment.\",\n \"Ash White Flies For each treatment, a 2.5 cm diam.\",\n \"leaf disc was mounted on a moistened Whatman #2 filter paper lining the bottom of petri dishes.\",\n \"Adult white flies were anaesthetised using carbon dioxide and 30-50 adults were transferred onto leaf discs.\",\n \"Serial dilutions of 0.00546, 0.0220, 0.0894 & 0.1788% concentration of E. cloeziana oil extract were prepared using distilled water containing 200 ppm Triton X100.Three replicates were treated for each concentration.\",\n \"Five-mL aliquots were applied to each petri dish using a Potter Spray Tower.\",\n \"After treatment, petri dishes were left to dry and then covered with muslin netting.\",\n \"Mortality assessment was assessed 24 h after white flies had been transferred to petri dishes.\",\n \"Drug Store Beetle One mL of each concentration of E. cloeziana oil extract in pure acetone was uniformly dispensed on 90 mm diam.\",\n \"Whatman No 2 filter paper.\",\n \"The latter was left for 1 h to air dry before being used to line the lid of a 90 mm diam.\",\n \"petri dish.\",\n \"Between 10-15 mixed adults were then transferred into the petri dish, which was sealed with Parafilm™.\",\n \"Mortality assessment was carried out 24 h after sealing the petri dish.\",\n \"Snails Two methods were used to assess the efficacy of E. cloeziana oil extract against snails.\",\n \"In the first method different age groups of adult snails were dipped directly into a solution of 0.5% concentration of E. cloeziana oil extract in distilled water containing 200 ppm Triton X10.Snails were dipped for 10 seconds and thereafter immediately filtered in a sieve.\",\n \"The treated snails were divided in three 500 mL kilner jars containing French bean leaves as a food source.\",\n \"The control was carried out using 0% E. cloeziana oil extract.\",\n \"Mortality was assessed 24 h after treatment.\",\n \"Allowing the snails to crawl on a E. cloeziana oil extract contaminated surface carried out the second method.\",\n \"Two mL of 0.08% concentration of E. cloeziana oil extract in pure acetone were dispensed in 500 mL kilner jars which were rotated to uniformly cover all inner surfaces until dry.\",\n \"Six different age groups of snails were put into each jar along with plant material for food and covered with muslin netting held by a rubber band.\",\n \"Three replicates of each treatment (0.0 & 0.08% concentration) were carried out.\",\n \"Mortality was assessed 24 h after releasing the snails inside the jars.\",\n \"The above bioassay results are summarised in Table 8.TABLE 8 SUMMARY OF E. CLOEZIANA OIL EXTRACT EFFICACY AGAINST VARIOUS TARGET PESTS Method of Target Pest LD50 (95% CL) LD95 (95% CL) application Remarks TSM 0.07 0.14 Potter spray Knockdown effect (0.06-0.08) (0.12-0.16) tower; Aliquot observed 2 h after applied 2.5 mL treatment TSM <0.01 E. cloeziana oil 100% mortality at extract + paraffin 0.01% oil TRM 0.02 0.04 Potter spray tower GHT 0.10 0.125 Potter spray tower Aphid 0.08 0.30 Potter spray tower American 157.27 μg/cm2 E. cloeziana oil cockroach residues on filter nymph paper Ant <0.0075 E. cloeziana oil 100% mortality at residues on filter 0.0075% when 1 mL paper applied as residue on filter paper Termite <0.0075 E. cloeziana oil 100% mortality at residues on filter 0.0075% when 1 mL paper applied as residue on filter paper DBM 1st 0.09 0.20 5 mL Potter spray instar larva tower Housefly 69.8 μg/cm2 130.87 μg/cm2 E. cloeziana oil adult residues on kilner jar wall Adult 0.00387 0.00694 Residues on Apparently very mosquitoes (0.00328-0.00451) (0.00596-0.01051) kilner jar wall rapid knock down.\",\n \"Ash white 0.01773 0.0.03728 Potter spray Assessed 4 h after fly (0.01179-0.03446) (0.02657-0.08337) tower, 2.5 mL treatment aliquot Drug store 0.03548 0.37275 Self Mortality assessed beetle contaminating by 24 h after releasing walking on filter adults.\",\n \"paper Honeybees 0.12 0.40 Potter spray tower Knockdown effect Aliquot applied 5 mL observed 2 h after treatment Snails ˜0.084 E. cloeziana oil Three mL acetone residues on kilner solution for each of jar wall 3 replicates.\",\n \"Snails crawled on the wall on dried residues.\",\n \"Snails 100% Dipping method Three replicates mortality in 0.5% Potted Plant Investigations The efficacy of E. cloeziana oil extract against TSM was Her assessed under greenhouse conditions.\",\n \"French bean plants were grown in 15 cm diam.\",\n \"plastic pots in an insecticide-free glasshouse maintained at 27° C., RH 65% and natural light.\",\n \"Plants were used when they reached the two true-leaf stage (ie., before trifoliate leaves appeared).\",\n \"One plant was maintained in each pot (i.e., two leaves/pot).\",\n \"Twenty gravid TSM females were then transferred with a fine brush to the upper surface of each leaf.\",\n \"Mites were left to settle for 4 h before treatment, during which time they usually settled on the lower leaf surfaces.\",\n \"E. cloeziana oil extract at a concentration of 0.07% as well as a blank control were prepared using the same procedures described above for the laboratory bioassays.\",\n \"A 400 mL hand sprayer was used to apply the pesticide evenly to all aerial surfaces of the plants, to run-off.\",\n \"Each treatment was replicated four times.\",\n \"The mortality was assessed 24 h after treatment.\",\n \"The results were recorded as mean percent mortality with standard deviation.\",\n \"These results revealed that the mean percent mortality in E. cloeziana oil extract treatment and control were 92.19±6.39 and 0.25±0.46, respectively.\",\n \"In summary, E. cloeziana oil extract was efficacious against all pests tested namely TSM, TRM, GHT, aphids, termites, houseflies, American cockroaches, whitefooted ants adult mosquitoes, ash whitefly, drug store beetle and snails.\",\n \"It was also toxic to honeybees.\",\n \"Example 8 E. cloeziana Oil Extract as a Fumigant An investigation was undertaken to determine the fumigation action of E. cloeziana oil extract against arthropods.\",\n \"The test organisms used were termites, Nasutitermes walkeri.\",\n \"A Whatman No 2 filter paper (90 mm diam.)\",\n \"was immersed in distilled water five seconds and left to drain excess water before placing it on the bottom of a 90 mm diam.\",\n \"petri dish, to provide moisture for termite workers during the experimental period.\",\n \"A 1.0% E. cloeziana oil solution was prepared in pure acetone.\",\n \"One mL was uniformly spread on a second filter paper, which was allowed to air dry on a sheet of aluminium foil.\",\n \"It was then placed under the lid of the petri dishes containing the moist filter paper on their base.\",\n \"Twenty uniform worker termites were transferred to the moist base of each test petri dish, which was subsequently covered with its lid containing the treated filter paper, and the dishes were then sealed with Parafilm™.\",\n \"A similar series of control treatments was also prepared, using acetone only, for comparison.\",\n \"Five replicates were used in each treatment.\",\n \"The termites did not move to the dry top surface and remained on the water-moistened filter paper lining the base of the petri dishes throughout the experimental period.\",\n \"Mortality was assessed 5 h after termite release.\",\n \"The results revealed that E. cloeziana oil extract has highly significant fumigant effects on termites.\",\n \"One hundred percent mortality was recorded in all replicates of the E. cloeziana oil treatment whereas no mortality occurred in any of the control (acetone only) replicates.\",\n \"Example 9 Efficacy of Purified β-Triketones Against TSM The efficacy of purified β-triketones against TSM was investigated using the TSM bioassay described in Example 8.All β-triketones tested demonstrated a high level of activity against TSM (see Table 9).\",\n \"TABLE 9 EFFICACY OF β-TRIKETONES AGAINST TSM Sample No.\",\n \"wt (mg) Chemical Name LD50 and 95% CL LD95 and 95% CL 1 109.2 Agglomerone 0.15 (0.11-021) 0.33 (0.22-1.06) 2 99.5 Grandiflorone 0.04 0.13 3 165.0 Dehydroangustione 0.36 (0.33-0.41) 0.69 (0.61-0.81) 4 107.1 Angustione 0.22 (0.21-0.24) 0.35 (0.31-0.40) 5 67.9 Jensenone No direct mortality occurred in any concentration tested (0.05-0.4%) within 24 h. However, all treated TSM were unable to move normally and continued to convulse until they commenced to die 72 h after treatment.\",\n \"There was no recovery.\",\n \"6 110.1 Tasmanone 0.055 0.150 7 101.8 Flavesone 0.020 (0.006-0.043) 0.0876 (0.076-0.114) 8 77.0 Leptospermone 0.037 0.169 9 113.4 Isoleptospermone 0.043 (0.027-0.057) 0.071 (0.058-0.109) 10 141.1 Platyphyllol 0.070 0.23 Example 10 Efficacy of Purified β-Triketones Against GHT The efficacy of purified β-triketones against GHT was investigated using the GHT bioassay described in Example 8.All β-triketones tested demonstrated a high level of activity against GHT (see Table 9).\",\n \"All β-triketones except jensenone caused 100% mortality on GHT at a concentration of 0.3% when 5 mL aliquots were applied with a Potter spray tower.\",\n \"The latter β-triketone did not cause direct mortality within 24 h, but caused behavioural effects at all concentrations tested.\",\n \"Convulsion and lack of movement were consistently observed and 60.0% mortality was recorded 72 h after application in the 0.4% treatment.\",\n \"Examples 11 Ready-to-use miticide spray - I Ingredient Parts E. cloeziana extract 0.1 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Solvent: e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 5 Tannic acid 1.0 Carrier e.g.\",\n \"water 92.9 Examples 12 Concentrated natural emulsifiable concentrate spray (4.4%) - I Ingredient Parts E. cloeziana extract 4.4 Pyrethrins 7.4 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 14.7 polyoxyethylenesorbitan, organosilicate Solvent: e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 73.5 Example 12 Concentrated emulsifiable concentrate spray (44%) Ingredient Parts E. cloeziana extract 22.0 Platyphyllol (natural or synthetic) 22.0 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 34.0 polyoxyethylenesorbitan, organosilicate Solvent: e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 22 Example 13 Natural ready to use insecticide spray Ingredient Parts E. cloeziana extract 0.3 Lavender oil 1.0 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Solvent e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 40 Carrier: Water 57.7 Example 14 Oil-based natural spray concentrate Ingredient Parts E. cloeziana extract 10.0 Petroleum oil 89.0 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Example 15 Concentrated emulsifiable concentrate spray (10%) Ingredient Parts E. cloeziana extract 10.0 Permethrin 10.0 Piperonyl butoxide 28.0 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 31.0 polyoxyethylenesorbitan, organosilicate Solvent e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 21 Example 16 Molluscidal dust Ingredient Parts E. cloeziana extract 2 Anti-caking agent (e.g.\",\n \"silica gel) 2 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 3 polyoxyethylenesorbitan, organosilicate Inert carrier (talc, kaolin, diatomaceous earth) 93 Example 17 Aerosol insecticidal and acaricidal spray Ingredient Parts E. cloeziana extract 1.0 Piperonyl butoxide 0.9 Propellent hydrocarbon 98.1 Example 18 Repellent Ingredient Parts E. cloeziana extract 19.5 Citronella oil 29.1 Phthalic acid dibutyl ester 29.1 N-octyl bicycloheptene dicarboxamide 22.3 Examples 19 Ready-to-use miticide spray - II Ingredient Parts Melaleuca cajeputi extract 0.2 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Solvent: e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 5.0 Tannic acid 1.0 Carrier e.g.\",\n \"water 92.8 Examples 20 Concentrated natural emulsifiable concentrate spray (4.4%) - II Ingredient Parts Melaleuca cajeputi extract 8.8 Pyrethrins 7.4 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 14.7 polyoxyethylenesorbitan, organosilicate Solvent: e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 69.1 Examples 21 Ready-to-use miticide spray - III Ingredient Parts 99% Tasmanone 0.09 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Solvent: e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 5 Tannic acid 1.0 Carrier e.g.\",\n \"water 92.91 Examples 22 Concentrated natural emulsifiable concentrate spray (4.4%) - III Ingredient Parts 99% Tasmanone 4.0 Pyrethrins 7.4 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 14.7 polyoxyethylenesorbitan, organosilicate Solvent: e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 73.9 Examples 23 Ready-to-use miticide spray - IV Ingredient Parts 99% Platyphyllol 0.09 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan, organosilicate Solvent: e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 5 Tannic acid 1.0 Carrier e.g.\",\n \"water 92.91 Examples 24 Concentrated natural emulsifiable concentrate spray (4.4%) - IV Ingredient Parts 99% Platyphyllol 4.0 Pyrethrins 7.4 Emulsifier: e.g.\",\n \"t-octylphenoxypolyethoxyethanol, 14.7 polyoxyethylenesorbitan, organosilicate Solvent: e.g.\",\n \"ethyl alcohol, isopropyl alcohol etc 73.9 Example 25 Intra-Specific Crosses for Imparting Pest Resistance to a Pest Susceptible Plant Controlled and wild pollination within Eucalyptus and other important commercial Myrtaceae are thoroughly addressed in the CRC for Sustainable Production Forestry Symposium on.\",\n \"Hybrid Breeding and Genetics—Controlled Pollination of Eucalypts on 12th April, 2000 and published as the proceedings of that symposium.\",\n \"Genetic Pollution from Farm Forestry (Potts et al., 2001) deals more specifically with intra and inter species crosses occurring within the Myrtaceae.\",\n \"Using the protocols described in the above publications, an intra-specific Eucalyptus cross breeding technique will employ the following protocol.\",\n \"This begins with the selection of supreme individuals as parent stock.\",\n \"Within a selected parent stock having superior pest resistant characteristics (e.g., E. cloeziana) male and female trees are identified for cross-pollination experiments.\",\n \"Pollen is harvested from the male trees and either stored or directly transferred to the female trees if flowering is synchronous.\",\n \"Emasculation is undertaken to preclude extraneous pollination occurring and flowers are often bagged as a further precaution.\",\n \"Seed set and subsequent embryo development proceeds over the ensuing 12-24 months and F1 seeds are collected at full maturation.\",\n \"Seeds are then germinated to produce seedlings that are subjected to detailed analysis to assess the transfer of traits from parent to progeny.\",\n \"If the F1 progeny show a desirable mix of phenotypic traits these progeny can be used to vegetatively propagate the new variety.\",\n \"If the F1 progeny show some improvement in the selected phenotypic traits, but further improvement is required selected F1 progeny can be back-crossed either within the F1 progeny or with one of the parent trees to produce an F2 progeny using the methodology outlined above.\",\n \"This iterative process can be continued ad infinitum until the desired characteristics are achieved.\",\n \"Example 26 Inter-Specific Crosses for Imparting Pest Resistance to a Pest Susceptible Plant Inter-species hybridisation in the wild, which is a common phenomenon within the subgenera of the Myrtaceae, have been recorded at almost 40% in the Eucalypts, 33% in Angophora 19% in Corymbia and 19% in Symphomyrtus.\",\n \"Hybridisation between the major subgenera may also occur (Potts et al., 2001).\",\n \"For example, Eucalyptus camaldulensis displays 14 natural hybrid crosses including E. camaldulensis XE.\",\n \"robusta, E. camaldulensis X E. alba, E. camaldulensis X E. cladocalyx, E. camaldulensis X E. bigalaterita, E. camaldulensis X E. tereticornis, E. camaldulensis X E. blakelyi, E. camaldulensis X E. dwyeri, E. camaldulensis X E. rudis, E. camaldulensis X E. ovata, E. camaldulensis X E. bridgesiana, E. camaldulensis X E. viminalis, E. camaldulensis X E. largiflorens, E. camaldulensis X E. melliodora, E. camaldulensis X E. leucoxylon; 15 manipulated hybrids including E. camaldulensis X E. diversicolor, E. camaldulensis X E. grandis, E. camaldulensis X E. botryoides, E. camaldulensis X E. cladocalyx, E. camaldulensis X E. tereticornis, E. camaldulensis X E. blakelyi, E. camaldulensis X E. urophylla, E. camaldulensis X E. macarthurfi, E. camaldulensis X E. exerta, E. camaldulensis X E. maidenii, E. camaldulensis X E. viminalis, E. camaldulensis X E. globulus, F. canialdulensis X E. gunnii, E. camaldulensis X E. laevopinea, E. camaldulensis XE.\",\n \"fastigata.\",\n \"In another example Eucalyptus globulus displays 15 natural hybrids including E. globulus XE.\",\n \"barberi, E. globulus XE.\",\n \"brookeriana, E. globulus XE.\",\n \"ovata, E. globulus XE.\",\n \"kitsoniana, E. globulus XE.\",\n \"goniocalyx, E. globulus XE.\",\n \"nortonii, E. globulus X E. cypellocarpa, E. globulus X E. pseudoglobulus, E. globulus X E. bicostata, E. globulus XE.\",\n \"johnstonii, E. globulus XE.\",\n \"viminalis, E. globulus XE.\",\n \"cordata, E. globulus XE.\",\n \"rubida, E. globulus XE.\",\n \"urnigera, E. globulus XE.\",\n \"perriniana and 13 successful manipulated hybrids including E. globulus X E. urophylla, E. globulus X E. grandis, E. globulus XE.\",\n \"robusta, E. globulus XE.\",\n \"pellita, E. globulus XE.\",\n \"longifolia, E. globulus X E. loxophloeba, E. globulus X E. camaldulensis, E. globulus X E. dunnii, E. globulus XE.\",\n \"nitens, E. globulus XE.\",\n \"maidenii, E. globulus XE.\",\n \"bicostata, E. globulus X E. viminalis, E. globulus XE.\",\n \"gunnii.\",\n \"In another example Eucalyptus grandis displays 4 natural hybrids including E. grandis XE.\",\n \"urophylla, E. grandis XE.\",\n \"robusta E. grandis X E. pellita E. grandis XE.\",\n \"terreticornis and 14 manipulated hybrids including E. grandis X E. urophylla E. grandis XE.\",\n \"botryoides E. grandis XE.\",\n \"pellita E. grandis XE.\",\n \"alba E. grandis XE.\",\n \"terreticornis E. grandis XE.\",\n \"camaldulensis E. grandis XE.\",\n \"dunnii E. nitens X E. maidenii E. grandis X E. globulus E. grandis X E. gunnii E. grandis X E. pulverulenta E. grandis X E. leucoxylon, E. grandis X E. resinifera.\",\n \"In another example the Corymbia henryi/variegata/maculata/citriodora complex displays 14 natural hybrids including C. citriodora X C catenaria, C. citriodora X C. variegata, C. citriodora X C. maculata, C. maculata X C. gummifrea, C. maculata X C. intermedia, C. maculata X C. citriodora, C. maculata X C. variegata, C. variegata X C. bloxsomeri, C. variegata X C. watsoniana, C. variegata X C. citriodora, C. variegata X C. maculata, C. Henryi X C. torelliana, C henryi X C. variegata and onemanipulated hybrid C. torelliana x c. In one further example Eucalyptus cloeziana displays only 1 natural hybrid E. cloeziana X E. acmenoides.\",\n \"The disclosure of every patent, patent application, and publication cited herein is hereby incorporated herein by reference in its entirety.\",\n \"The citation of any reference herein should not be construed as an admission that such reference is available as “Prior Art” to the instant application.\",\n \"Throughout the specification the aim has been to describe the preferred embodiments of the invention without limiting the invention to any one embodiment or specific collection of features.\",\n \"Those of skill in the art will therefore appreciate that, in light of the instant disclosure, various modifications and changes can be made in the particular embodiments exemplified without departing from the scope of the present invention.\",\n \"All such modifications and changes are intended to be included within the scope of the appended claims.\",\n \"BIBLIOGRAPHY Bignall, C. M., Dunlop, P. J., Brophy, J. J. and Fookes, C. J. R. (1997).\",\n \"Volatile Leaf Oils of some South-western and Southern Australian Species of the Genus Eucalyptus (Series I).\",\n \"Part XIV.\",\n \"Subgenus Monocalyptus.\",\n \"Flav Frag.\",\n \"Journal, 12, 177-183.Boland, D. and J. Brophy (1993).\",\n \"Essential Oils of the Eucalyptus and Related Genera: Search for Chemical Trends.\",\n \"Bioactive Volatile Compounds from Plants.\",\n \"P, Teranishi, R. G. Buttery and H. Sugisawa.\",\n \"Washington D.C., American Chemical Society.\",\n \"525: 72-87.Brophy, J. J. and D. J. Boland (1990).\",\n \"“Leaf Essential Oil of Two Chemotypes of Eucalyptus cloeziana F. Muell.” Journal of Essential Oil Research 2(Mar/Apr): 87-90.Brophy, J. J., R. J. Goldsack, et al.\",\n \"(1995).\",\n \"“Leaf Oils of the Genus Backhousia (Myrtaceae).” Journal of Essential Oil Research 7(May/Jun): 237-254.CRC for Sustainable Production Forestry.\",\n \"Symposium on Hybrid Breeding and Genetics—Controlled Pollination of Eucalypts, Noosa Australia, 12th April, 2000 Hellyer, R. (1968).\",\n \"“The Occurrence of O-Triketones in the Steam-Volatile Oils of some Myrtaceous Australian Plants.” Aust.\",\n \"J. Chem.\",\n \"21(11): 2825-2828.Herron G A, Beatie G A C, Parkes R A & Barchia I.\",\n \"1995.Potter spray tower bioassay of selected citrus pests to petroleum spray oil.\",\n \"Journal of Australian Entomological Society 34: 225-263.Potts, B M, Barbour, R C and Hingston, A H, 2001.Genetic Pollution from Farm Forestry.\",\n \"Rural Industries Research and Development Corporation Publication (No 01/114).\",\n \"Southwell, I.\",\n \"A. and J. J. Brophy (2000).\",\n \"“Essential oil isolates from the Australian Flora.\",\n \"Part 3.” Journal of Essential Oil Research 12: 267-278.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10477057"}}},{"rowIdx":999,"cells":{"text":{"kind":"list like","value":[["Production of pure molybdenum oxide from low grade molybdenite concentrates","High purity ammonium dimolybdate or molybdenum oxide is produced by the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.","The process entails nearly complete oxidation of the sulfide minerals while optimizing the process chemistry and autoclave conditions to solubilize as little of the molybdenum values as possible.","The autoclave discharge 12 is then subjected to a leaching step, either an alkaline leach 50, 400 or ammonium leach 250 process, before or after a liquid/solid separation step 20, 220, 410.","The solution is then subjected to (a) filtration 60, 410, solvent extraction 70, 440, crystallization 90, 450, and calcination 120, 480 or (b) filtration 260, 280, crystallization 290, and calcination 320 to produce a product suitable for chemical-grade molybdenum oxide 125, 325, 485."],["1-54.","(canceled) 55.A method of producing high purity ammonium dimolybdate from low grade molybdenite concentrates comprising: a) forming an aqueous slurry of said low grade molybdenite concentrates; b) oxidizing said slurry in an atmosphere containing free oxygen at a pressure of at least about 50 p.s.i.","and at a temperature of at least about 200° C. and thereafter producing a first discharge with greater than about 99% of the molybdenum in said low grade molybdenite concentrates oxidized and greater than about 80% of molybdenum values insoluble; c) filtering said first discharge to produce a first liquid filtrate containing soluble molybdenum values and a first solid filter cake containing the insoluble molybdenum values; d) leaching said first solid filter cake with an alkaline solution to produce a second discharge wherein greater than about 98% of the insoluble molybdenum values are solubilized; e) filtering said second discharge to produce a second liquid filtrate and a second solid filter cake; f) recovering the molybdenum values from said second liquid filtrate by solvent extraction with an organic solvent to produce a first liquor by (1) contacting said second liquid filtrate with said organic solvent to form a two-phase mixture and simultaneously reducing the pH level in said two-phase mixture such that the molybdenum values in said second liquid filtrate are extracted into said organic solvent; and (2) stripping said organic solvent with aqueous ammonia to recover the molybdenum values; g) crystallizing said first liquor containing the extracted molybdenum values to produce crystals and a second liquor; and h) recovering said high purity ammonium dimolybdate from said crystals.","56.The method of claim 55, wherein the organic solvent contains a secondary amine.","57.The method of claim 56, wherein the organic solvent contains ditridecyl amine.","58.The method of claim 55, wherein the reduced pH level in said two-phase mixture ranges from about 4.0 to about 4.5.59.The method of claim 55, wherein the reducing is accomplished by adding hydrated sulfuric acid to said two-phase mixture.","60.The method of claim 55, wherein ammonium hydroxide is used during the stripping to recover the molybdenum values.","61.The method of claim 55, wherein the pH level during the stripping is maintained at about 9.0.62.A method of producing high purity ammonium dimolybdate from low grade molybdenite concentrates comprising: a) forming an aqueous slurry of said low grade molybdenite concentrates; b) oxidizing said slurry in an atmosphere containing free oxygen at an pressure of at least about 50 p.s.i.","and at a temperature of at least about 200° C. and thereafter producing a first discharge with greater than about 99% of the molybdenum in said low grade molybdenite concentrates oxidized and greater than about 80% of molybdenum values insoluble; c) separating and filtering said first discharge to produce a first liquid filtrate containing soluble molybdenum values and a first solid filter cake containing the insoluble molybdenum values; d) leaching said first solid filter cake with an ammoniacal solution to produce a second discharge wherein greater than about 98% of the insoluble molybdenum values are solubilized; e) filtering said second discharge to produce a second liquid filtrate and a second solid filter cake; f) aging said second liquid filtrate; g) crystallizing said second liquid filtrate to produce crystals and a first liquor; and h) recovering said high purity ammonium dimolybdate from said crystals; wherein the method further comprises subjecting said first liquid filtrate produced in step (c) to a solvent extraction process to recover molybdenum values, said solvent extraction process comprising contacting said first liquid filtrate with an organic solvent to form a two-phase mixture and simultaneously reducing the pH level in said two-phase mixture such that the molybdenum values in said first liquid filtrate are extracted into said organic solvent, and stripping said organic solvent to produce a second liquor containing the recovered molybdenum values.","63.The method of claim 62, wherein the organic solvent contains a secondary amine.","64.The method of claim 63, wherein the organic solvent contains ditridecyl amine.","65.The method of claim 62, wherein the reduced pH level in said two-phase mixture ranges from about 4.0 to about 4.5.66.The method of claim 62, wherein the reducing is accomplished by adding hydrated sulfuric acid to said two-phase mixture.","67.The method of claim 62, wherein sulfuric acid is used during the stripping to recover the molybdenum values.","68.The method of claim 62, wherein the pH level during the stripping is maintained at about less than 3.0.69.The method of claim 62, further comprising recycling a portion of said second liquor back to the aqueous slurry in step (a).","70.The method of claim 62, further comprising subjecting said second liquor to a cementation process to recover copper values, said cementation process comprising adding iron to said second liquor and mixing to produce a first solution, filtering said first solution to produce a third liquid filtrate and a third solid filter cake, and then recovering said copper values from said third solid filter cake.","71.A method of producing high purity ammonium dimolybdate from low grade molybdenite concentrates comprising: a) forming an aqueous slurry of said low grade molybdenite concentrates; b) oxidizing said slurry in an atmosphere containing free oxygen at an pressure of at least about 50 p.s.i.","and at a temperature of at least about 200° C. and thereafter producing a first discharge with greater than about 99% of the molybdenum in said low grade molybdenite concentrates oxidized and greater than about 80% of molybdenum values insoluble; c) separating and filtering said first discharge to produce a first liquid filtrate containing soluble molybdenum values and a first solid filter cake containing the insoluble molybdenum values; d) leaching said first solid filter cake with an ammoniacal solution to produce a second discharge wherein greater than about 98% of the insoluble molybdenum values are solubilized; e) filtering said second discharge to produce a second liquid filtrate and a second solid filter cake; f) aging said second liquid filtrate and adding a reagent selected from the group consisting of iron molybdate, ammonium sulfide, sulfide compounds, and ferric sulfate to said second liquid filtrate; g) crystallizing said second liquid filtrate to produce crystals and a first liquor; and h) recovering said high purity ammonium dimolybdate from said crystals.","72.A method of producing high purity ammonium dimolybdate from low grade molybdenite concentrates comprising: a) forming an aqueous slurry of said low grade molybdenite concentrates; b) oxidizing said slurry in an atmosphere containing free oxygen at an pressure of at least about 50 p.s.i.","and at a temperature of at least about 200° C. and thereafter producing a first discharge with greater than about 99% of the molybdenum in said low grade molybdenite concentrates oxidized and greater than about 80% of molybdenum values insoluble; c) leaching said first discharge with an alkaline solution to produce a second discharge wherein greater than about 98% of the insoluble molybdenum values are solubilized; d) separating and filtering said second discharge to produce a liquid filtrate containing soluble molybdenum values and a solid filter cake containing the insoluble molybdenum values; e) recovering the molybdenum values from said liquid filtrate by solvent extraction with an organic solvent to produce a first liquor by (1) contacting said liquid filtrate with said organic solvent to form a two-phase mixture and simultaneously reducing the pH level in said two-phase mixture such that the molybdenum values in said liquid filtrate are extracted into said organic solvent; and (2) stripping said organic solvent to recover the molybdenum values; f) crystallizing said first liquor containing the extracted molybdenum values to produce crystals and a second liquor; and g) recovering said high purity ammonium dimolybdate from said crystals.","73.The method of claim 72, wherein the organic solvent contains a secondary amine.","74.The method of claim 73, wherein the organic solvent contains ditridecyl amine.","75.The method of claim 72, wherein the reduced pH level in said two-phase mixture ranges from about 4.0 to about 4.5.76.The method of claim 72, wherein the reducing is accomplished by adding hydrated sulfuric acid to said two-phase mixture.","77.The method of claim 72, wherein ammonium hydroxide is used during the stripping to recover the molybdenum values.","78.The method of claim 72, wherein the pH level during the stripping is maintained at about 9.0."],["<SOH> BACKGROUND OF THE INVENTION <EOH>Extraction of molybdenum from molybdenite-containing materials by an aqueous process has been the subject of study for over 50 years.","In 1952, E. S. Usataya 1 reported on the oxidation of molybdenite in water solutions.","He found that in neutral, acidic, or weakly alkaline solutions the decomposition products precipitate on the molybdenite surface and protect the molybdenite from further oxidation.","He found that strong bases and strong oxidizing agents impede the formation of the protective layers.","Increasing temperature accelerated the oxidation rate in alkaline solutions, but up to 60° C. had no effect in acidic solutions.","1 Usataya, E. S., “Oxidation of molybdenite in water solutions,” Zapiski Vsesoyuz Mineral Obshschestva, v 81, 298-303 (1952).","A Japanese patent 2 was issued for oxygen pressure oxidation (POX) of molybdenite in 1962.The example in this patent leached a 55.5% Mo, 36.4% S, and 4.4% Cu concentrate at 9% solids at 200° C. and 200 atmospheres oxygen.","The molybdic acid precipitate that formed during leaching was dissolved using ammonia for recovery of an ammonium molybdate.","2 Sada, Koji, “Extraction of molybdenum,” Japanese patent 15.207('62), assigned to Awamura Mining Co., Ltd.","In another process disclosure 3 , alkali hydroxide and alternatively ammonium hydroxide was added continuously to the aqueous solution to neutralize the acid as it formed and maintain the pH at 7-12.Other authors 4 postulated the formation of a molybdenum-iron heteropoly complex that decomposes as the acid concentration increases.","It also may be a ferrous complex that decomposes as the soluble iron is oxidized to ferric.","3 Hallada, Calvin J., et al., “Conversion of molybdenum disulfide to molybdenum oxide,” German patent 2,045,308 (1971).","4 Mel'nikov, B. S. and Shapiro, K. A., “Water-autoclave decomposition of molybdenite raw material,” Protsessy Poluch.","Rafinirovaniya Tugoplavkikh Met.","(1975) 113-120, 253-260.Early applications of molybdenum solvent extraction utilized tertiary amine to extract the molybdenum solubilized by sodium hydroxide leaching of roasted molybdenite calcines.","Secondary amines and quaternary ammonium compounds extract anionic molybdenum using a similar chemistry.","Amines also were used for extraction of molybdenum from molybdenite roaster scrubber solutions.","Molybdenum is an impurity in many uranium ores.","When uranium ores are acid leached, some molybdenum reports to the acid leach solution.","The tertiary amines readily available during the 1950's and early 1960's tended to have an amine-molybdenum complex with poor solubility in aliphatic diluents (kerosene).","Several Russian researchers worked with acid leaching of oxide ores.","In many cases the acidity was sufficient for most of the molybdenum to be in a cationic form.","Therefore, the cation exchanger (di, 2, ethylhexyl phosphoric acid (DEHPA)) received much study regarding the recovery of molybdenum from complex acid solutions.","Karpacheva et al.","5 determined that in acid solutions the molybdenum was not present as the simple molybdenyl cation but, the molybdenum was present as polymeric cations.","The co-extraction of iron is a major problem when using DEHPA.","The authors noted that in a nitric acid system, the acid concentration needs to equal or exceed 3 molar to prevent significant iron extraction.","Other authors 6 reported on the benefit of modifiers in reducing the iron coextraction, e.g.","tributyl phosphate, dibutyl butyl phosphonate.","5 Karpacheva, S. M. et al., “Extraction of molybdenum and iron (III) by di-2-ethylhexyl hydrogen phosphate,” Russian Journal of Inorganic Chemistry, V 12, 7, p 1014-1016 (1967).","6 Chiola, Vincent, “Separation of molybdenum values from tungsten values by solvent extraction,” U.S. Pat.","No.","3,607,008 (1971).","Palant et al.","7 made a detailed study of the extraction of molybdenum by DEHPA.","The solutions studies were prepared by dissolving MoO 3 in sulfuric acid, hydrochloric acid, or nitric acid solutions.","Palant, A.","A. et. al.","“Extraction of molybdenum (VI) with bis(2-ethylhexyl) hydrogen phosphate from an acidic medium,” Inst.","Metall.","im.","Baikova, Moskow, USSR, Report deposited (1979) pp.","1-19.Amine exchange has also received much study during the past 40 to 50 years.","The difficulties presented by the poor solubility of the amine-molybdenum complex were addressed by using aromatic diluents.","MacInnis et al.","8 used tri-n-capryl amine (Alamine 336) with the aromatic diluent #28 9 .","The authors discuss amine extraction of a complex sulfate-bearing anion.","At pH values of 3 and higher, they determined that the ion exchange type mechanism shown in Equation I below predominates.","8 MacInnis, M. B., Kim, T. K., and Laferty, J. M., “The use of solvent extraction for the production and recovery of high-purity ammonium paramolybdate from normal alkali molybdate solution,” First Intl Conf on Chemistry and Uses for Molybdenum, p. 56-58 (1973).","9 Aromatic diluent #28 is a solvent from 1960 available from Missouri Solvents & Chemicals.","The solvent had a boiling range of 165 to 193° C., a flash point of 122° F., a Kauri butanol value of 73, and was 74% aromatics.","in-line-formulae description=\"In-line Formulae\" end=\"lead\"?","2MoS 2 +6H 2 O+9O 2 →2H 2 MoO 4 ↓+4H 2 SO 4 (I) in-line-formulae description=\"In-line Formulae\" end=\"tail\"?","At pH values below 3, they postulate that the following occurs along with Equation I. in-line-formulae description=\"In-line Formulae\" end=\"lead\"?","n (R 3 NH.HSO 4 )+(Mo x O y H z ) n .","(SO 4 ) m →(R 3 NH) n .","(Mo x O y H z ) n .","(HSO4) n .","(SO 4 ) m (II) in-line-formulae description=\"In-line Formulae\" end=\"tail\"?","Equation II infers that (Mo x O y H z ) n .","(SO 4 ) m is not ionized and Maclnnis postulates some cation transfer.","This postulation is based on the fact that 35 S tagged sulfur was found to transfer both from the organic to the aqueous phase and from the aqueous to the organic phase.","Litz found in 1970, that tris, tridecyl amine could be used successfully for molybdenum solvent extraction with an aliphatic diluent.","There still was potential for formation of insoluble molybdenum-amine complexes, but the molybdenum-tris tridecyl amine complex's solubility in the diluent was much higher than with other tertiary amines.","Tris tridecyl amine in an aliphatic diluent was used in a number of pilot circuits for molybdenum solvent extraction from roaster scrubber solutions and to recover byproduct from uranium leach solutions, but it may never have been used in a commercial circuit.","The transfer of sulfate from strongly acid solutions is a problem with using amines.","Also, the amines are relatively nonselective and will transfer silicon, phosphorus, and arsenic probably as heteropoly compounds.","The coextraction of silicon and subsequent solids precipitation during stripping has been a major problem during other studies.","The silicon problem was addressed by filtration of the first stripping stage mixture prior to advancing to the settler.","Sulfate transfer was high because the solvent could not be fully loaded with molybdenum, i.e., to avoid diluent-insoluble molybdenum-amine complexes, the sulfate transfer was large.","Efficient recovery of chemical-grade ammonium dimolybdate (ADM) requires high purity feed solutions containing 200 to 230 g Mo per liter.","Impurities in the solution must be removed to avoid inclusion in the ADM. Impurities, that form hydroxides or sulfides, can be removed by additions or pH-control.","Other impurities will build up and unless the mother liquor is bled from the crystallization will report to the ADM.","Typical molybdenum solvent extraction systems acidulate the feed solution, if necessary, prior to contact with the extractant in the mixer.","Generally this means that the extractant is converted to the bisulfate form by acid in the feed solution and then the desired anion exchanges with the bisulfate.","When molybdenum is acidulated there is potential for localized high acid concentrations that can form sulfate-bearing molybdenum species.","It is an object of the present invention to provide an integrated process for producing high purity ammonium dimolybdate or molybdenum oxide through a process that includes the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.","It is a further object of the present invention to provide an improved molybdenum pressure oxidation process which produces a high purity product at reduced capital and operating costs.","It is a further object of the present invention to provide an improved solvent extraction method which rejects sulfate and metallic impurities by extracting the molybdenum in an ionic form that contains no sulfate."],["<SOH> SUMMARY OF THE INVENTION <EOH>The objects set forth above as well as further and other objects and advantages of the present invention are achieved by the present invention now described in summary fashion and with further examples below in preferred embodiments of the practice of the invention.","The present invention provides a process of producing a high purity ammonium dimolybdate or molybdenum oxide through the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.","The process entails oxidizing the molybdenite concentrates or intermediates in an autoclave operating at greater than 50 p.s.i.","oxygen overpressure, preferably between 80-120 p.s.i., at a temperature greater than 200° C., preferably between 210-220° C. to effect almost complete oxidation of the concentrate while optimizing the process chemistry and autoclave conditions to solubilize as little of the molybdenum values as possible.","A method of maximizing the insoluble molybdenum values is disclosed in U.S Patent Application entitled “Autoclave Control Mechanisms for Pressure Oxidation of Molybdenite” which is incorporated by reference herein (and a copy of which is provided at Appendix A hereto).","The resulting autoclave discharge has greater than 99% of the molybdenum concentrates oxidized and greater than 80% of the molybdenum values insoluble.","The autoclave discharge is then subjected to an alkaline leaching of the POX residue using sodium carbonate and sodium hydroxide.","More than 99% of the molybdenum dissolves.","The molybdenum in this alkaline solution is recovered readily using a secondary amine solvent, di,tridecyl amine (DTDA).","The molybdenum is loaded into the organic phase at 4.0 to 4.5 pH.","The molybdenum-loaded organic is stripped with ammonium hydroxide to produce solutions suitable for recovery of chemical-grade ADM and ultimately chemical-grade molybdenum oxide.","Alternatively, the autoclave discharge may be subjected to an ammoniacal leaching of the POX residue.","More than 99% of the molybdenum dissolves.","Most of the cosolubilized impurities are precipitated from the ammoniacal leach solution.","The solution is evaporated to crystallize chemical grade ADM. An additional route is provided when the leach solution contains more sulfate than is desirable for crystallization of chemical-grade ADM, producing a product suitable for technical grade ADM and ultimately technical grade molybdenum oxide.","Alternatively, the POX residue may be subjected to an alkaline leach with sodium carbonate and sodium hydroxide before a liquid-solid separation step such that all of the molybdenum is soluble and the copper and iron transfer to the solids.","More than 98% of the molybdenum is transferred to the filtrate from this neutralization.","The molybdenum is readily recoverable using the DTDA solvent extraction process.","The ammoniacal strip solutions from the DTDA extraction are suitable for recovery of chemical-grade ADM and ultimately chemical-grade molybdenum oxide.","Cementation by scrap iron can be used to readily recover the copper either from the raffinate produced from molybdenum solvent extraction of the POX leach solution or directly from the POX leach solution.","Gold and silver values transfer to the final leached solid residue produced by each embodiment and are recyclable to a copper smelter.","Most of the rhenium, arsenic, and phosphorus are dissolved regardless of the leaching conditions.","Other objects, features and advantages of the invention will be apparent from the following description of preferred embodiments thereof, including illustrative non-limiting examples of the practice of the process."],["FIELD OF THE INVENTION The present invention relates to the process of producing ammonium dimolybdate for conversion to pure molybdenum oxide or other pure chemicals from molybdenite concentrates and more particularly to a process of producing chemical grade ammonium dimolybdate for conversion to chemical grade molybdenum oxide through a process that includes the pressure oxidation of low-grade molybdenite concentrates.","BACKGROUND OF THE INVENTION Extraction of molybdenum from molybdenite-containing materials by an aqueous process has been the subject of study for over 50 years.","In 1952, E. S. Usataya1 reported on the oxidation of molybdenite in water solutions.","He found that in neutral, acidic, or weakly alkaline solutions the decomposition products precipitate on the molybdenite surface and protect the molybdenite from further oxidation.","He found that strong bases and strong oxidizing agents impede the formation of the protective layers.","Increasing temperature accelerated the oxidation rate in alkaline solutions, but up to 60° C. had no effect in acidic solutions.","1 Usataya, E. S., “Oxidation of molybdenite in water solutions,” Zapiski Vsesoyuz Mineral Obshschestva, v 81, 298-303 (1952).","A Japanese patent2 was issued for oxygen pressure oxidation (POX) of molybdenite in 1962.The example in this patent leached a 55.5% Mo, 36.4% S, and 4.4% Cu concentrate at 9% solids at 200° C. and 200 atmospheres oxygen.","The molybdic acid precipitate that formed during leaching was dissolved using ammonia for recovery of an ammonium molybdate.","2 Sada, Koji, “Extraction of molybdenum,” Japanese patent 15.207('62), assigned to Awamura Mining Co., Ltd.","In another process disclosure3, alkali hydroxide and alternatively ammonium hydroxide was added continuously to the aqueous solution to neutralize the acid as it formed and maintain the pH at 7-12.Other authors4 postulated the formation of a molybdenum-iron heteropoly complex that decomposes as the acid concentration increases.","It also may be a ferrous complex that decomposes as the soluble iron is oxidized to ferric.","3 Hallada, Calvin J., et al., “Conversion of molybdenum disulfide to molybdenum oxide,” German patent 2,045,308 (1971).","4 Mel'nikov, B. S. and Shapiro, K. A., “Water-autoclave decomposition of molybdenite raw material,” Protsessy Poluch.","Rafinirovaniya Tugoplavkikh Met.","(1975) 113-120, 253-260.Early applications of molybdenum solvent extraction utilized tertiary amine to extract the molybdenum solubilized by sodium hydroxide leaching of roasted molybdenite calcines.","Secondary amines and quaternary ammonium compounds extract anionic molybdenum using a similar chemistry.","Amines also were used for extraction of molybdenum from molybdenite roaster scrubber solutions.","Molybdenum is an impurity in many uranium ores.","When uranium ores are acid leached, some molybdenum reports to the acid leach solution.","The tertiary amines readily available during the 1950's and early 1960's tended to have an amine-molybdenum complex with poor solubility in aliphatic diluents (kerosene).","Several Russian researchers worked with acid leaching of oxide ores.","In many cases the acidity was sufficient for most of the molybdenum to be in a cationic form.","Therefore, the cation exchanger (di, 2, ethylhexyl phosphoric acid (DEHPA)) received much study regarding the recovery of molybdenum from complex acid solutions.","Karpacheva et al.5 determined that in acid solutions the molybdenum was not present as the simple molybdenyl cation but, the molybdenum was present as polymeric cations.","The co-extraction of iron is a major problem when using DEHPA.","The authors noted that in a nitric acid system, the acid concentration needs to equal or exceed 3 molar to prevent significant iron extraction.","Other authors6 reported on the benefit of modifiers in reducing the iron coextraction, e.g.","tributyl phosphate, dibutyl butyl phosphonate.","5 Karpacheva, S. M. et al., “Extraction of molybdenum and iron (III) by di-2-ethylhexyl hydrogen phosphate,” Russian Journal of Inorganic Chemistry, V 12, 7, p 1014-1016 (1967).","6 Chiola, Vincent, “Separation of molybdenum values from tungsten values by solvent extraction,” U.S. Pat.","No.","3,607,008 (1971).","Palant et al.7 made a detailed study of the extraction of molybdenum by DEHPA.","The solutions studies were prepared by dissolving MoO3 in sulfuric acid, hydrochloric acid, or nitric acid solutions.","Palant, A.","A. et. al.","“Extraction of molybdenum (VI) with bis(2-ethylhexyl) hydrogen phosphate from an acidic medium,” Inst.","Metall.","im.","Baikova, Moskow, USSR, Report deposited (1979) pp.","1-19.Amine exchange has also received much study during the past 40 to 50 years.","The difficulties presented by the poor solubility of the amine-molybdenum complex were addressed by using aromatic diluents.","MacInnis et al.8 used tri-n-capryl amine (Alamine 336) with the aromatic diluent #289.The authors discuss amine extraction of a complex sulfate-bearing anion.","At pH values of 3 and higher, they determined that the ion exchange type mechanism shown in Equation I below predominates.","8 MacInnis, M. B., Kim, T. K., and Laferty, J. M., “The use of solvent extraction for the production and recovery of high-purity ammonium paramolybdate from normal alkali molybdate solution,” First Intl Conf on Chemistry and Uses for Molybdenum, p. 56-58 (1973).","9 Aromatic diluent #28 is a solvent from 1960 available from Missouri Solvents & Chemicals.","The solvent had a boiling range of 165 to 193° C., a flash point of 122° F., a Kauri butanol value of 73, and was 74% aromatics.","2MoS2+6H2O+9O2→2H2MoO4↓+4H2SO4 (I) At pH values below 3, they postulate that the following occurs along with Equation I. n(R3NH.HSO4)+(MoxOyHz)n.(SO4)m→(R3NH)n.(MoxOyHz)n.(HSO4)n.(SO4)m (II) Equation II infers that (MoxOyHz)n.(SO4)m is not ionized and Maclnnis postulates some cation transfer.","This postulation is based on the fact that 35S tagged sulfur was found to transfer both from the organic to the aqueous phase and from the aqueous to the organic phase.","Litz found in 1970, that tris, tridecyl amine could be used successfully for molybdenum solvent extraction with an aliphatic diluent.","There still was potential for formation of insoluble molybdenum-amine complexes, but the molybdenum-tris tridecyl amine complex's solubility in the diluent was much higher than with other tertiary amines.","Tris tridecyl amine in an aliphatic diluent was used in a number of pilot circuits for molybdenum solvent extraction from roaster scrubber solutions and to recover byproduct from uranium leach solutions, but it may never have been used in a commercial circuit.","The transfer of sulfate from strongly acid solutions is a problem with using amines.","Also, the amines are relatively nonselective and will transfer silicon, phosphorus, and arsenic probably as heteropoly compounds.","The coextraction of silicon and subsequent solids precipitation during stripping has been a major problem during other studies.","The silicon problem was addressed by filtration of the first stripping stage mixture prior to advancing to the settler.","Sulfate transfer was high because the solvent could not be fully loaded with molybdenum, i.e., to avoid diluent-insoluble molybdenum-amine complexes, the sulfate transfer was large.","Efficient recovery of chemical-grade ammonium dimolybdate (ADM) requires high purity feed solutions containing 200 to 230 g Mo per liter.","Impurities in the solution must be removed to avoid inclusion in the ADM. Impurities, that form hydroxides or sulfides, can be removed by additions or pH-control.","Other impurities will build up and unless the mother liquor is bled from the crystallization will report to the ADM.","Typical molybdenum solvent extraction systems acidulate the feed solution, if necessary, prior to contact with the extractant in the mixer.","Generally this means that the extractant is converted to the bisulfate form by acid in the feed solution and then the desired anion exchanges with the bisulfate.","When molybdenum is acidulated there is potential for localized high acid concentrations that can form sulfate-bearing molybdenum species.","It is an object of the present invention to provide an integrated process for producing high purity ammonium dimolybdate or molybdenum oxide through a process that includes the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.","It is a further object of the present invention to provide an improved molybdenum pressure oxidation process which produces a high purity product at reduced capital and operating costs.","It is a further object of the present invention to provide an improved solvent extraction method which rejects sulfate and metallic impurities by extracting the molybdenum in an ionic form that contains no sulfate.","SUMMARY OF THE INVENTION The objects set forth above as well as further and other objects and advantages of the present invention are achieved by the present invention now described in summary fashion and with further examples below in preferred embodiments of the practice of the invention.","The present invention provides a process of producing a high purity ammonium dimolybdate or molybdenum oxide through the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.","The process entails oxidizing the molybdenite concentrates or intermediates in an autoclave operating at greater than 50 p.s.i.","oxygen overpressure, preferably between 80-120 p.s.i., at a temperature greater than 200° C., preferably between 210-220° C. to effect almost complete oxidation of the concentrate while optimizing the process chemistry and autoclave conditions to solubilize as little of the molybdenum values as possible.","A method of maximizing the insoluble molybdenum values is disclosed in U.S Patent Application entitled “Autoclave Control Mechanisms for Pressure Oxidation of Molybdenite” which is incorporated by reference herein (and a copy of which is provided at Appendix A hereto).","The resulting autoclave discharge has greater than 99% of the molybdenum concentrates oxidized and greater than 80% of the molybdenum values insoluble.","The autoclave discharge is then subjected to an alkaline leaching of the POX residue using sodium carbonate and sodium hydroxide.","More than 99% of the molybdenum dissolves.","The molybdenum in this alkaline solution is recovered readily using a secondary amine solvent, di,tridecyl amine (DTDA).","The molybdenum is loaded into the organic phase at 4.0 to 4.5 pH.","The molybdenum-loaded organic is stripped with ammonium hydroxide to produce solutions suitable for recovery of chemical-grade ADM and ultimately chemical-grade molybdenum oxide.","Alternatively, the autoclave discharge may be subjected to an ammoniacal leaching of the POX residue.","More than 99% of the molybdenum dissolves.","Most of the cosolubilized impurities are precipitated from the ammoniacal leach solution.","The solution is evaporated to crystallize chemical grade ADM. An additional route is provided when the leach solution contains more sulfate than is desirable for crystallization of chemical-grade ADM, producing a product suitable for technical grade ADM and ultimately technical grade molybdenum oxide.","Alternatively, the POX residue may be subjected to an alkaline leach with sodium carbonate and sodium hydroxide before a liquid-solid separation step such that all of the molybdenum is soluble and the copper and iron transfer to the solids.","More than 98% of the molybdenum is transferred to the filtrate from this neutralization.","The molybdenum is readily recoverable using the DTDA solvent extraction process.","The ammoniacal strip solutions from the DTDA extraction are suitable for recovery of chemical-grade ADM and ultimately chemical-grade molybdenum oxide.","Cementation by scrap iron can be used to readily recover the copper either from the raffinate produced from molybdenum solvent extraction of the POX leach solution or directly from the POX leach solution.","Gold and silver values transfer to the final leached solid residue produced by each embodiment and are recyclable to a copper smelter.","Most of the rhenium, arsenic, and phosphorus are dissolved regardless of the leaching conditions.","Other objects, features and advantages of the invention will be apparent from the following description of preferred embodiments thereof, including illustrative non-limiting examples of the practice of the process.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is a flow diagram of one embodiment of the process of the present invention in which the POX residue is subjected to an alkaline leach; FIG.","2 is a flow diagram of another embodiment of the process of the present invention in which the POX residue is subjected to an ammoniacal leach; and FIG.","3 is a flow diagram of another embodiment of the process of the present invention in which the POX residue is subjected to an alkaline leach prior to liquid-solid separation.","DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS A process of producing a pure ammonium dimolybdate or molybdenum oxide through the pressure oxidation of low-grade molybdenite concentrates is provided in a first embodiment.","Referring to FIG.","1, the process begins with the pressure oxidation 10 of low grade molybdenite concentrate or intermediate concentrates at greater than 50 p.s.i.","oxygen overpressure, preferably between 80-120 p.s.i., at a temperature greater than 200° C., preferably between 210-220° C. while optimizing the process chemistry and autoclave conditions to solubilize as little of the molybdenum values as possible.","This can be accomplished by a high degree of oxidation of sulfide sulfur in the autoclave and acceptably high H2SO4 concentrations.","Treated water and recycle wash water from the autoclave filter is added to dilute the feed to the autoclave in order to control the temperature in the autoclave.","Recycling a portion of the autoclave discharge slurry 12 back to the autoclave increases the acidity and ferric level in the autoclave.","The recycled autoclave discharge aqueous accelerates the leaching rate, thus reducing the reactor size, and the additional acid limits the soluble molybdenum to about 15% of the total.","Recycling some of the autoclave slurry 12 back to the autoclave also provides seeding material to increase the size of the MoO3 particles to improve filterability.","The resulting autoclave discharge has greater than 99% of the molybdenum concentrates oxidized and greater than 80% of the molybdenum values insoluble.","The autoclave discharge slurry is then filtered 20 and the filtrate 22 is treated by scrap iron 24 to precipitate the copper.","The cementation process 30 begins by adding the iron 24 to the liquid 22 at room temperature.","The scrap iron reacts with the filtrate according to the following equations.","Fe+2Fe3+→3Fe2+ (III) Fe+Cu2+→Cu+Fe2+ (V) The recovered copper precipitate will be filtered, washed, and sent to a smelter 32.The chemistry of tail stream 34 from iron cementation 30 is then adjusted and molybdenum is precipitated using metallic iron powder 36 in a stirred tank reactor 40.This product 42 is recycled back to the autoclave feed.","If the cementation process 30 is not used, then the filtrate 22 advances directly to the molybdenum precipitation process 40.The insoluble molybdenum values 26 in the POX residue are present as hydrated and anhydrous molybdenum oxides and are readily soluble in alkaline solutions.","The autoclave discharge filter cake 26 is then repulped in water and a soda ash solution 28 is added to dissolve the molybdenum 50.Sodium carbonate alone dissolves most of the molybdenum at final pH values below 7.0 but a significant amount of iron was also found to dissolve.","Sodium hydroxide is then added to increase the pH level to about 9.0 to 10.0, eliminating the soluble iron and producing a leach slurry having low arsenic, phosphorus, and selenium content.","The amount of soluble silicon is variable.","The discharge 52 from the soda ash/caustic leach 50 is then filtered and washed 60.The filter cake 62 is sent to the smelter for recovery of precious metals.","The filtrate 64 is sent to solvent extraction 70.The sodium molybdate solution 64 produced by the alkaline leach 50 is subjected to an amine solvent extraction 70 of the molybdenum.","The molybdenum extraction mixers can be operated to produce a two-phase mixture having either the aqueous- or organic-phase continuous.","The organic phase contains 10% DTDA.","Controlling the acidity in the mixers at 4.0 to 4.5 pH by the direct addition of hydrated sulfuric acid (65% H2SO4 or so) minimizes the transfer of arsenic, phosphorus, selenium, silicon, and sulfate.","The molybdenum-loaded organic then is washed with a weak acid solution and/or water, and then is stripped with an ammonium hydroxide solution.","Concentrated ammonium hydroxide or anhydrous, gaseous ammonia is added to control the pH in stripping at about 9.0.After stripping, the organic is water-washed to remove entrained strip solution and the aqueous is advanced as make-up water to stripping.","The ideal solvent molybdenum extraction rejects sulfate and metallic impurities.","The most efficient method of rejecting sulfate is to extract the molybdenum in an ionic form that contains no sulfate.","Typical solvent extraction systems acidulate the feed solution, if necessary, prior to contact with the extractant in the mixer.","Generally this means that the extractant is converted to the bisulfate form and then the desired anion exchanges with the bisulfate.","In addition, when molybdenum-bearing solutions are acidulated there is potential for localized high acid concentrations that can form sulfate-bearing molybdenum specie.","The molybdenum ion present in neutral to alkaline solutions is the molybdate ion.","On mild acidulation to 4.0 to 4.5 pH, the molybdenum will form large polymolybdate ions that are readily extractable.","If the acidulation of the feed solution is done simultaneously in the mixer to produce a two-phase mixture, the extractant quickly exchanges the polymolybdate species before the molybdenum is further acidulated to form a sulfate-bearing specie.","Typically the metallic impurities will not form large polyions at the target pH values and, if they do, the polymolybdates will displace them along with any bisulfate ions from the amine.","Therefore, small amounts of sulfate and silicon are the only impurities that follow the molybdenum.","Raffinate 72 from the solvent extraction is sent to the disposal tank for neutralization and disposal with the other effluents from the plant.","The pregnant liquor 74 from the solvent extraction 70 advances to the aging and purification tanks 80 to allow silica to coagulate.","Iron molybdate and/or ammonium sulfide 82 may be added to the aging tanks to reduce the amount of trace impurities.","The aged and purified solution 84 is then sent to a crystallizer 90 where solid ammonium dimolybdate (ADM) is produced.","The crystals are recovered in a centrifuge 100.The vapor 92 from the crystallizer 90 is condensed to recover the ammonia 110 for use in the solvent extraction process 70.The filtrate 102 from the centrifuge is recycled back to the crystallizer 90.As an option, a bleed stream 94 from the crystallizer circuit can be taken to recover rhenium values in a rhenium extraction process 130 and/or the bleed stream 94 can be neutralized with soda ash and steam-stripped to recover and recycle the ammonia.","The ammonia-free solution then is returned to the alkaline leach 50.In the rhenium extraction process 130, the rhenium is extracted from the crystallizer mother liquor bleed in ion exchange columns.","Stripping is done with ammonia.","The strip solution is sent to a crystallizer to recover the rhenium as solid ammonium perrhenate.","The crystals 104 from the crystallizer centrifuge 100 are transported to a dryer/calciner 120.The dried/calcined material may then be sent to packaging.","The resulting material is suitable for chemical grade ADM or chemical grade molybdenum oxide 125.Table 1 below shows the requirements for chemical grade molybdenum oxide in comparison to technical grade molybdenum oxide.","TABLE 1 Technical Grade Chemical Grade MoO3 Spec MoO3 Spec Al, max.","ppm — 10 Ca, ppm — 5 Co, ppm — 10 Cr, ppm — 5 Cu, ppm 4500 5 Fe, ppm — 10 K, ppm — 80 Mg, ppm — 5 Mn, ppm — 10 Ni, ppm — 5 P, ppm 200 10 Pb, ppm 300 10 S, ppm 300 — Si, ppm 20,000-40,000 10 Sn, ppm — 10 Ti, ppm — 5 V, ppm — 10 Zn, ppm — 10 Zr, ppm — 10 Referring to FIG.","2, in another embodiment of the present invention, the autoclave discharge 12 is centrifuged and washed and then repulped, filtered and washed again 220.The purpose is to obtain a molybdenum-containing filter cake 222 which is low in soluble impurities such as silicon and sulfate.","The centrate 224, which contains almost all of the copper and much of the iron from the feed, plus the balance of the molybdenum, is sent to the optional solvent extraction process 230 for molybdenum recovery.","The solvent extraction process is essentially the same as described in the first embodiment except the stripping is done with a concentrated sulfuric acid solution rather than an ammonium hydroxide solution.","During stripping, the pH is maintained at about less than 3.0.Recovered molybdenum values are recycled back to the autoclave feed.","Raffinate 232 from the solvent extraction 230 advances to copper recovery 240, followed by molybdenum precipitation by scrap iron.","If the solvent extraction process 230 is not used, then the solution advances directly to copper recovery 240.The filter cake 222 is contacted with ammonium hydroxide solution and anhydrous ammonia 250 rather than the soda ash/caustic leach solution 50 of the first embodiment.","The solid molybdic oxide dissolves to form soluble ammonium molybdate.","The residue after dissolution is the insoluble portion.","The dissolver slurry 252 advances to a continuous filter 260.Some filtrate 262 may be recycled to reduce the solids concentration in the dissolver 250.Both iron molybdate and ammonium sulfide 254 may be added to either the dissolver 250 or the aging tanks 280.These two reagents precipitate trace impurities.","The filtrate 262 from the dissolver filter 260 goes to a control tank with a recycle stream back to the dissolver to maintain proper specific gravity and pH.","The filtrate from the control tank then goes to the aging and purification tanks 280 to allow silica to coagulate.","The aged and purified solution 282 is then sent to a crystallizer 290 where solid ammonium dimolybdate (ADM) is produced.","The crystals are recovered in a centrifuge 310.The vapor 292 from the crystallizer 290 is condensed to recover the ammonia 300 for use in the dissolver 250.The majority of the mother-liquor filtrate 312 from the centrifuge 310 is recycled to the crystallizer 290.The crystals from the crystallizer centrifuge 310 are transported to a dryer/calciner 320.The dried/calcined material results in chemical grade ADM or chemical grade molybdenum oxide 325.A part of the mother-liquor filtrate is used as a continuous bleed stream 294 from the crystallizer circuit to control impurities in the mother liquor in the crystallizer 290.This bleed stream 294 is sent back to the dissolver 250 to precipitate the build-up of impurities.","If there is a build-up of impurities which can not sufficiently be precipitated in the dissolver/aging circuit, a part of the bleed stream 296 is sent to a second crystallizer 330 to produce an impure ADM.","The crystals from the second crystallizer 330 are recovered in a centrifuge 340.The vapor 332 from the second crystallizer 330 is condensed to recover the ammonia 300 for use in the dissolver 250.The crystals from the second crystallizer centrifuge 340 are transported to a dryer/calciner 350.The calcined material results in technical grade ADM or technical grade molybdenum oxide 355, which can be sold or recycled back 252 to the dissolver 250 or the autoclave 10.Referring to FIG.","3, in another embodiment of the present invention, the autoclave discharge 12 is subjected to the sodium carbonate/sodium hydroxide leaching process 400 of the first embodiment prior to liquid-solid separation.","The leaching discharge 402 is then thickened, centrifuged and washed, repulped, filtered on a continuous pressure filter and rewashed 410.The solids 412 containing the copper and gold are sent to the smelter 422 for recovery.","The filtrate 414 containing the molybdenum and rhenium is sent to a rhenium extraction process 430.The rhenium is extracted from the alkaline filtrate in ion exchange columns.","Stripping is done with ammonia.","The strip solution is sent to a crystallizer to recover the rhenium as solid ammonium perrhenate.","The molybdenum-bearing solution 432 from rhenium extraction 430 is pumped to the solvent extraction process 440 previously described in the first embodiment.","The raffinate 442 from the solvent extraction 440 is sent to the plant effluent.","The pregnant liquor 444 is sent to the crystallization process 450 previously described in the first embodiment producing chemical grade ADM or chemical grade molybdenum oxide 485.EXAMPLES The invention is now further disclosed with reference to the following non-limiting Examples.","Example 1 Cementation Process After pressure oxidation of the molybdenite concentrate in the autoclave and the solid-liquid separation of the slurry, the resulting acidic liquid is treated by cementation.","The major constituents of the liquid are approximately: Mo, g/l 10-16 Cu, g/l 8-11 Fe, g/l 8-11 H2SO4, g/l 100 The cementation process is conducted at room temperature and begins by adding to the liquid about 1.0 gram Fe per gram Fe present (as Fe3+) in the liquid and about 2.0 gram Fe per gram Cu present in the liquid.","The slurry is mixed for 10-15 minutes and then filtered.","The solids contain the recovered Cu-values.","The filtrate is then treated for the recovery of Mo.","The pH of the filtrate is increased to about 1.05-1.2 by the addition of Na2CO3 or NaOH and the temperature of the filtrate is increased to about 40-65° C. An additional 1.5-2.1 gram Fe per gram Mo present in the liquid is then added.","The slurry is mixed for 15-30 minutes and filtered.","The filtrate should have a clear yellow/greenish color.","A recycle stream, which feeds solids or slurry back to the mix-tank is needed to promote the filterability of the Mo-cake.","The solids contain the recovered Mo-values.","The filtrate can be neutralized with lime and discarded.","The mass distribution in % of the elements is shown in Table 2.TABLE 2 Starting Ele- acidic Cu-step, Cu-step, Mo-step, Mo-step, ment liquid Solids Liquid Solids Liquid Notes Mo 100 1-1.5 98.5-99 98.4 0.07-0.24 Cu 100 98.5-99.99 0-1.5 <0.01 0.01 Si 100 15 85 As 100 85 15 Se 100 100 0 (* Re 100 50 50 (* SO4 100 2 6-20 (* P 35 65 (** Mg 30 70 (** Al 10 90 (** Notes: (* = The liquid phase is not analyzed, but based on the synthetic solutions it is assumed that the distribution of the elements are as is indicated.","(** = The distribution only indicates the relative distribution between the two solid phases.","Example 2 Alkaline Leach A number of tests were performed to attempt optimization of the leaching conditions while minimizing the formation of bicarbonates.","The alkaline leaching data are summarized in Table 3.Leaching at pH values below 7.0 using sodium carbonate demonstrated almost complete solubilization of the molybdenum but significant co-solubilization of iron, probably as a carbonate complex, was also present.","Leaching with sodium hydroxide did not solubilize much iron.","Therefore, tests to optimize leaching cost and efficiency evaluated an initial leach with sodium carbonate to 6 pH, followed by sodium hydroxide to 9 pH or only using sodium hydroxide.","Molybdenum extraction in these tests (see Table 3, Leach No.","CL-1 to CL-6) exceeded 98%.","The leach solutions contained 43 to 79 g/l Mo and silicon was the only impurity of significance.","The reagent requirement averaged about 1.1 lb Na2CO3 and 0.7 lb NaOH per lb of molybdenum dissolved.","Example 3 Alkaline Leach Solvent Extraction A number of tests were performed to determine the optimum conditions for solvent extracting molybdenum from the alkaline leach solutions.","The molybdenum solvent extraction studies used an organic containing 10% di, tridecyl amine, 5% decyl alcohol, and 85% Escaid 110.The results of these studies are summarized in Tables 4A-4C.","The initial tests evaluated the effect of temperature in the pH range of 2.0-2.7.Temperature had no significant effect on the molybdenum extraction.","The extraction was very efficient with raffinates containing from 1 to 40 mg molybdenum/liter from feed solutions containing 63 to 70 g/l (greater than 99.9% transfer).","The distribution coefficients in the first contact, TABLE 3 ALKALINE LEACHING OF POX LEACH RESIDUE MOLYBDENUM EXTRACTION AND LEACH SOLUTION IMPURITIES Dissolution Leach Temp Na2CO3 NaOH Soluble Na2CO3 NaOH Filtrate Filtrate Impurities, mg/l on a 200 g/l Mo basis No.",": ° C. to ?","pH to ?","pH Mo g/g Mo g/g Mo Mo, g/l Cu Fe As K P Re Mg Si Se RL-30 50 13 95.6% 0.95 65.3 18 13 37 172 RL-31 50 13 91.9% 0.84 93.6 4 235 RL-32 50 6.8 99.6% 1.31 33.4 24 72 RL-33 50 5.2 99.9% 0.68 31.6 1500 25 468 RL-34 50 3.5 94.5% 0.38 32.2 1500 25 200 RL-35 50 4.3 99.5% 0.39 48.4 1300 8 314 RL-36 50 6.5 98.5% 0.59 76 1040 10 550 RL-37 50 8.05 99.6% 1.00 81.9 <3 7 10 100 RL-38 50 9.54 99.8% 0.99 83.1 <3 5 14 140 RL-39 50 10 95.6% 1.00 74.8 <3 5 5 45 11 27 350 CL-1 50 5.6 9.2 98.0% 1.04 0.72 67.1 12 1500 CL-2 50 5.6 9.4 98.6% 1.04 0.76 58.6 14 2400 CL-3 50 10 98.1% 1.18 74.2 11 890 CL-4 50 9.6 98.4% 1.03 78.8 10 470 CL-5 50 9.0 9.3 98.9% 1.52 0.18 42.9 10 325 CL-6 50 6.9 9.5 99.0% 1.16 0.33 71.1 39 6 124 28 28 TABLE 4A SOLVENT EXTRACTION SUMMARY ALKALINE LEACH SOLUTIONS Impurities Test Temp Mo Content, g/l H2SO4 In strip, mg/l per 200 g/l Mo No.","pH ° C. Organic Aqueous g/g Mo Si As P Re SO4 1210 35 Feed 82 110 8 4.8 Loaded Stage 41.5 55 <2 6 1.5 First Contact 27.8 28 0.8 <2 6 0.7 2.5 Raffinate Stage 0.22 <0.001 <2 4 0.05 First Strip Stage, 8.5 pH 3 81 260 25 6.4 Second Strip Stage, 9.5 0.6 7 <55 230 10 1220 Feed 34 2.5 Loaded Stage 45.6 30 4 2.5 First Contact 22.5 0.02 0.41 <2 2.5 Raffinate Stage 0.01 <0.001 <2 First Strip Stage, 8.5 pH 4.1 86 9 107 Second Strip Stage, 9.0 0.3 11 <35 1230 25 Feed 45 76 2 2.5 Loaded Stage 45 47 <2 4 2.5 First Contact 29.6 0.08 <2 <2 2.5 Raffinate Stage 0.01 0.02 <2 4 First Strip Stage, 8.5 pH 9.7 65 215 6 Second Strip Stage, 9.0 1.5 23 70 <15 1410 35 Feed 83 275 6 1700 4.2 Loaded Stage 53.5 61 4.4 First Contact 16.6 0.13 First Strip Stage, 9.0 pH 1.0 183 164 u u 39 Second Strip Stage, 9.0 0.1 6 870 200 <30 370 Re 1420 35 Alkaline Feed 61 10 14 2 10 44 POX Leach Feed 11 80 60 54 26 16 4.4 Alkaline Contact 42.4 25 <2 8 <1 44 20 First POX Contact 7.3 0.33 50 22 28 3 <2 9.1 Strip of Alkaline Contact 4.3 104 31 510 310 200 173 Alkaline Raff Contact 22.7 0.39 <2 6 <1 <1 <1 9.0 Strip of Alkaline Raff Co 0 46 <9 26 9 <9 35600 TABLE 4B SOLVENT EXTRACTION SUMMARY ALKALINE LEACH SOLUTIONS Solvent: 10% DTDA 5% Decyl alcohol 85% Escaid 110 Impurites Test Temp Mo Content, g/l H2SO4 In Strip, mg/l per 200 g/l Mo No.","pH ° C. Organic Aqueous g/g Mo Si As P Re 1030 50 Feed 70 56 6 na 12 2.1 Loaded Stage 46.0 1.29 <2 6 0.4 4 1.4 Raffinate Stage 0.06 <0.001 <2 6 0.05 <2 First Strip Stage, 6.5 pH 21 39 <10 <10 5 41 Second Strip Stage, 9.0 p 8 23 313 35 42 <17 1040 50 Feed 70 56 6 na 12 2.6 Loaded Stage 41.6 7.62 <2 6 0.3 4 2.6 Raffinate Stage 0.12 0.016 <2 <2 0.2 <2 First Strip Stage, 6.5 pH 14 43 <9 <9 7 37 Second Strip Stage, 9.0 p 0.5 20 380 <20 25 <20 1070 35 Feed 65 56 6 2.6 Loaded Stage 43.3 0.32 <2 2.8 Raffinate Stage 0.05 0.11 <2 First Strip Stage, 8.5 pH 9.8 63 146 6 Second Strip Stage, 9.5 p 5.6 12 <30 <30 1080 20 Feed 63 56 6 2.7 Loaded Stage 40.9 0.85 2 2.7 Raffinate Stage 0.01 0.003 2 First Strip Stage, 8.5 pH 9.9 72 89 6 Second Strip Stage, 9.5 p 0.1 1.2 <300 <300 1090 35 Feed 70 56 6 12 2.1 Loaded Stage 46.5 0.35 <2 6 2 2.0 Raffinate Stage 0.009 0.001 <2 4 2 First Strip Stage, 8.5 pH 11.8 73 142 5 Second Strip Stage, 9.5 p 0.37 34 200 <5 1100 20 Feed 65 56 6 12 1.95 Loaded Stage 41 3.7 <2 6 2 2.00 Raffinate Stage 0.03 0.04 <2 4 <2 First Strip Stage, 8.5 pH 25.6 32 310 12 Second Strip Stage, 9.5 p 14.4 33 25 <10 TABLE 4C SOLVENT EXTRACTION SUMMARY ALKALINE LEACH SOLUTIONS Impurities Test Temp Mo Content, g/l H2SO4 In strip, mg/l per 200 g/l Mo No.","pH ° C. Organic Aqueous g/g Mo Si AS P Re SO4 1110 35 Feed 65 56 6 2.1 Loaded Stage, 120 seco 35.3 13 4 First Strip Stage, 8.5 pH 4.1 35 183 <10 Second Strip Stage, 9.5 2.9 1.3 <300 300 2.1 Loaded Stage, 60 secon 22.5 31 20 4 First Strip Stage, 8.5 pH 3.4 45 187 9 Second Strip Stage, 9.5 2.9 1.5 <250 <250 2.1 Loaded Stage, 30 secon 23.4 30 12 4 First Strip Stage, 8.5 pH 3.7 46 174 9 Second Strip Stage, 9.5 3.3 1.2 <300 <300 2.1 Loaded Stage, 15 secon 16.8 40 20 4 First Strip Stage, 8.5 pH 0 65 197 6 1150 50 Feed 85 110 8 12 3.0 Loaded Stage 41.5 22 <2 6 1 3.1 Raffinate Stage 0.04 0.07 <2 6 First Strip Stage, 8.5 pH 1.3 76 <5 11 16 42 470 Second Strip Stage, 9.5 0.3 2.8 <140 <140 <70 <140 1500 1160 50 Feed 82 110 8 12 3.52 Loaded Stage 39.3 23 <2 6 1 3.66 Raffinate Stage 0.01 0.04 <2 6 First Strip Stage, 8.5 pH 2.1 76 <5 11 13 63 320 Second Strip Stage, 9.5 0.6 4 <100 <100 <50 <100 1700 1170 35 Feed 82 110 8 12 2.5 Loaded Stage 40.7 20 <2 6 8 First Strip Stage, 8.5 pH 1.6 81 197 15 3.7 Second Strip Stage, 9.5 0.3 3.7 750 110 16 freshly stripped organic and feed, were as high as 133 (g/l Mo organic phase÷g/l Mo aqueous phase).","There was some transfer of silicon and arsenic.","The first strip stages contained up to 310 mg/l silicon and 12 mg/l arsenic when normalized to 200 g/l molybdenum.","The effect of contact time on the molybdenum and impurity transfer was also evaluated.","As the contact time was reduced from 120 to 15 seconds, the molybdenum transfer was reduced from 80% to 40%.","The ratios of silicon and arsenic to molybdenum in subsequent strip solutions did not change, indicating that varying contact time would not vary the amount of impurity transferred.","The transfer of sulfate to the strip solutions was also monitored.","The pH of the extraction contact was increased.","Increasing the pH of the contact reduced the amount of sulfate transferred.","The silicon transfer also was reduced in the higher pH contacts.","The amount of sulfate transferred to the strip ranged from 320 to 470 mg/l when normalized to 200 g/l molybdenum.","The effect of temperature (25, 35, and 50° C.) at pH levels of 2.5 to 3.5 was also evaluated.","Within these ranges, temperature and pH appeared to have no effect on the molybdenum transfer.","Phase separation was slightly faster at 50° C. The transfer of silicon was lower at 50° C. Sulfate transfer at 50° C. and 3.0 to 3.5 pH was low.","Strip solutions contained 320 to 470 mg/l SO4 on a 200 g/l molybdenum basis.","A batch contact test was done at the pH found most efficient for molybdenum transfer, 4.2-4.4.The first contact distribution coefficient was 133.The first contact aqueous phase contained 0.13 g/l Mo, indicating 99.8% extraction in the first contact.","The transfer of silicon was reduced significantly to 164 mg/l in the strip solution when normalized to 200 g/l Mo.","Sulfate transfer also was reduced to 39 mg/l on a 200 g/l Mo basis.","Countercurrent Solvent Extraction Two countercurrent extractions were performed and the data from these tests are summarized in Table 5.In both tests a single extraction contact was made.","The target pH in the extraction stage was 4.0 to 4.3.The actual pH values of the eight contacts ranged TABLE 5 COUNTERCURRENT SOLVENT EXTRACTION ALKALINE LEACH SOLUTIONS Test No: 1390 Organic: NaOH Leach Aqueous: 10% DTDA 55 g/l Mo 5% Decyl alcohol g/l Fe 85% Escaid 110 g/l H2SO4 mg/l Si Strip Prod Impurities on Loaded Stage 200 g/l Mo basis Contact Mo, g/l Mo, Si, As, P, pH Aqueous Organic g/l mg/l mg/l mg/l Cycle 1 2.3 0.037 217 180 6 Cycle 2 4.0 0.060 224 195 9 Cycle 3 4.3 0.49 221 360 17 Cycle 4 3.8 0.023 226 195 37 Test No: 1400 Organic: NaOH Leach Aqueous: 10% DEHPA 73 g/l Mo 5% Decyl alcohol 275 mg/l Si 85% Escaid 110 3 mg/l P 6 mg/l As 1700 mg/l SO4 Strip Prod Impurities on a Loaded Stage 200 g/l Mo basis Contact Mo, g/l Mo, Si, As, P, SO4, pH Aqueous Organic g/l mg/l mg/l mg/l mg/l Cycle 3.7 14.6 222 240 5 3 65 1 Cycle 4.1 12.9 228 720 5 1 23 2 Cycle 4.2 13.8 226 620 7 3 95 3 Cycle 4.1 14.5 22.7 220 645 5 2 79 4 from 2.3 to 4.3, with only one contact being at a pH lower than 3.7.In this pH range it is easy to add excess acid once the needs for species change were met.","Controlling the pH would not be a problem in a continuous circuit.","The loaded solvent was water-washed and stripped countercurrently with three stages of ammonium hydroxide.","Each test was operated for four cycles.","With countercurrent stripping it was possible to produce product strip solutions containing up to 228 g/l Mo.","The transfer of sulfate was low, 23 to 95 mg/l on a 200 g/l Mo basis.","The strip solutions were well below the target values of phosphorus and arsenic (1-3 and 5-7 mg/l respectively), but contained 240 to 720 mg/l silicon.","Example 5 Sulfate Removal from Pressure Oxidation Residue Two washing/re-pulping tests were performed to study removal of sulfate from the autoclave discharge.","The results of these tests are shown in Table 6.TABLE 6 Test 1 Test 2 Soluble SO4 per 200 g/l Mo when leached Initial filter cake 250 g/l 250 g/l After 2 displacement washes 12.3 g/l 5.3 g/l Cake after one repulp <4.8 g/l 0.73 g/l After 2 displacement washes <1.4 g/l 0.11 g/l Example 6 Ammoniacal Leaching A series of tests were performed to determine the optimum conditions for ammoniacal leaching of the autoclave residue.","The ammoniacal leaching data are summarized in Table 7.The first set of tests evaluated leaching POX residues by adding the wet cake to reagent ammonium hydroxide (nominally 500 g/l NH4OH).","The initial TABLE 7 DATA SUMMARY LEACHING OF AUTOCLAVE RESIDUE WITH AMMONIA Test 40 41 42 43 44 45 46 47 Temp 55 55 55 55 55 55 50 50 Pulp Density <43 <38 <79 <79 <79 <79 <82 <79 Stages 1 1 2 2 2 2 2 2 Hours 2 2 2 2 2 2 2 2 Final pH 9.4 9.0 9.0 8.5 9.4 8.8 9.0 9.1 Added iron molybdate no no no no yes yes yes yes Aged overnight no yes yes yes yes yes yes yes Feed, % Mo comp.","comp.","28.7 27.7 29.3 27.4 25.5 26.4 Residue, % Mo 0.46 0.64 0.16 0.09 0.10 0.25 0.29 0.68 Ammonia soluble, % 99.0 98.4 99.7 99.8 99.8 99.5 99.3 98.3 Filtrate, g/l Mo 31.8 73.7 178 142 158 182 184 181 Impurities per 200 g/l Mo Si 226 190 103 85 56 48 74 82 As <13 33 9 8.5 10 11 11 13 P na 5 6 7 7 8 Cu 19 16 4.5 8.5 11 12 14 14 Fe 132 na 5.6 7 4 3 1 1 K <1 <1 1 1 Mg 2 3 3 2 SO4 380 <330 1300 4240 ammonium hydroxide additions were 1.24 and 1.7 times stoichiometric to dissolve the molybdenum.","The mixtures then were heated to 55° C. for two hours and, if necessary, ammonium hydroxide was added to keep the pH above 9.0.The total ammonium hydroxide additions were 1.7 and 3.5 times stoichiometric to dissolve the molybdenum.","After the leaching period, the leach slurries were transferred to plastic bottles, sealed, and aged overnight at 50° C. The molybdenum dissolutions were both excellent at 99.7 and 99.8%.","The leach filtrates contained 178 and 142 g/l Mo.","The soluble impurities on a 200 g/l Mo basis, except for silicon were low at 85-103 mg/l Si, 8.5-9 mg/l As, 2-3 mg/l Mg, <1 mg/l K, 4.5-8.5 mg/l Cu, and 5.6-7 mg/l Fe.","The next set evaluated adding iron molybdate and sodium sulfide to the leach slurry prior to the aging period.","The initial ammonium hydroxide additions were 1.08 and 1.17 times stoichiometric to dissolve the molybdenum.","The mixtures then were heated to 55° C. for two hours and, if necessary, ammonium hydroxide was added to keep the pH above 9.0.The total ammonium hydroxide additions were 2.0 and 1.8 times stoichiometric to dissolve the molybdenum.","At 90 minutes iron molybdate was added to the leach slurry and at 105 minutes sodium sulfide was added to the leach slurry.","At 120 minutes, the leach slurries were transferred to plastic bottles, sealed, and aged overnight at 50° C. The molybdenum dissolutions were both excellent at 99.8 and 99.5%.","The leach filtrates contained 158 and 182 g/l Mo.","The soluble impurities on a 200 g/l Mo basis were low at 48-56 mg/l Si, 10-11 mg/l As, 2-3 mg/l Mg, 1 mg/l K, 6-7 mg/l P, 11-12 mg/l Cu, and 3-4 mg/l Fe.","The sulfate levels were 380 and less than 330 mg SO4/l.","The last set evaluated using anhydrous ammonia to adjust the pH after the initial pulping in ammonium hydroxide solution.","The initial ammonium hydroxide additions were 1.65 and 1.72 times stoichiometric to dissolve the molybdenum.","The mixtures then were heated to 55° C. for two hours and, if necessary, anhydrous ammonia was added to keep the pH above 9.0.At 90 minutes a small amount of iron molybdate cake was added to the slurry.","After the leaching period, the leach slurries were transferred to plastic bottles, sealed, and aged overnight at 55° C. The molybdenum dissolutions were both excellent at 99.3 and 98.3%.","The leach filtrate contained 184 and 181 g/l Mo.","The soluble impurities on a 200 g/l Mo basis were 74-82 mg/l Si, 11-13 mg/l As, 7-8 mg/l P, 14 mg/l Cu, and 1 mg/l Fe.","The sulfate levels in the leach solution were high, 1300 and 4240 mg/l SO4, even though the POX residue had been washed, repulped, refiltered and rewashed.","Example 7 Purification of Ammonia Leach Solutions Tests were performed to evaluate purification of leach solutions with high molybdenum concentration, 142-223 g/l Mo.","Data are summarized in Table 8.Additives evaluated included ferric sulfate, magnesium sulfate, iron molybdate, and aluminum molybdate.","Ferric sulfate and iron molybdate were effective in reducing the silicon by 50 to 70%.","Magnesium sulfate had little effect, although the final pH values were below the hydrolysis point of magnesium hydroxide.","Aluminum molybdate showed little effect.","The addition of iron molybdate successfully reduced the silicon to below the target concentration with each of the solutions.","The iron molybdate for the above tests was prepared by mixing solutions of ferric sulfate or ferric chloride with a sodium molybdate solution while controlling the pH at 1.6 to 1.9.Example 8 Total Soda Leach (TSL) Process A series of tests were performed to determine the optimum conditions for neutralizing the autoclave leach slurry prior to solid-liquid separation.","The data are summarized in Table 9.This process eliminates one liquid-solid separation step when compared with the alkaline leaching process.","Sodium carbonate was added to a fixed pH, 6.0 to 8.0, and then in some cases sodium hydroxide was added to pH 9.0.The data in Table 9 show that the final pH needs to be at least 9.0 for the molybdenum solubility to exceed 90%.","The quantities of soluble impurities in the neutralized solution were very low as shown in Table 10.Tests on leach slurries No.","164 and 165 focused on determining the lowest cost combination of sodium carbonate and sodium hydroxide that could be used for the neutralization.","Because of the formation of bicarbonate, using sodium carbonate is not as cost effective at higher pH values as sodium hydroxide.","The leaching-neutralization tests showed that more than 98% of the molybdenum would be soluble after partial TABLE 8 PURIFICATION OF AMMONIUM HYDROXIDE LEACH Test P-41 P-43 P-45 P-42 P-44 P-46 P-47 P-48 P-49 Temp 50 50 50 50 50 50 50 50 50 50 pH 9.0 7.4 7.5 7.3 9.0 7.4 7.4 7.4 8.0 8.5-9.0 9.5 8.5-9.0 8.9 Addition, g/l iron molybdate 28 g 40 g 20 g 10 g wet wet wet wet Ferric sulfate 10 14 Magnesium sulfate hyd 12 16 12 Aluminum molybdate Feed Filtrate Filtrate Filtrate Feed Filtrate Filtrate Filtrate Filtrate Feed Filtrate Feed Filtrate Molybdenum, g/l 142 132 100 155 178 170 134 168 157 184 180 181 180 Impurities per 200 g/l Mo Si 85 42 28 88 103 38 48 114 84 74 40 82 27 As 8 9 12 8 9 7 9 7 8 11 7 13 4 P 6 5 12 6 6 5 10 6 5 7 8 Cu 8 8 12 9 4 >4 >9 7 8 14 9 14 9 Fe 7 9 8 8 6 22 12 7 5 1 1 K 1 2 2 1 1 1 1 1 1 Mg 3 3 32 1400 2 2 15 1400 1300 SO4 1304 4243 TABLE 9 SUMMARY OF TSLP LEACH AND NEUTRALIZATION Leach Conc.","Filtrate, g/l Soluble Residue, S % Temp Na2CO3 NaOH Filtrate Residue, % Soluble No.",": Feed Mo Cu H2SO4 Mo, % AC A Res ° C. to ?","pH to ?","pH Mo, g/l Mo Cu Mo, % 157 1B 7.1 8.5 65 98.2 25 7.0 12.6 5.6 5.1 68 8.0 14.9 4.1 5.8 77 8.0 9.0 24.6 3.2 4.7 84 158 1B 14.7 6.7 44 99.7 25 7.0 8.4 8.6 4.3 45 8.0 14.8 4.8 4.7 74 8.0 9.0 23.8 2.3 4.6 87 160 1B 20.5 9.0 53 89.3 1.86 1.66 25 6.0 9.0 16.8 4.9 4.1 74 161 1B 5.1 7.1 73 99.3 0.81 0.01 25 6.0 9.0 17.0 2.2 4.9 92 162 4A 9.1 6.5 99.8 0.81 0.10 25 6.0 9.0 18.2 5.2 11.1 91 163 4A 6.1 8.8 99.9 0.10 0.12 25 6.0 9.0 19.0 3.8 11.3 93 164 2B 7.0 3.8 84 99.9 0.25 0.01 35 9.0 33.1 2.6 94.4 35 10.0 35.7 2.0 95.6 55 9.0 30.2 3.7 92.0 55 10.0 33.2 0.6 98.8 75 9.0 36.3 2.3 95.1 75 10.0 35.0 2.0 95.4 55 6.0 9.0 20.8 0.8 98.5 55 6.0 10.0 21.0 0.3 99.5 75 6.0 9.0 27.9 1.3 97.1 75 6.0 10.0 29.8 0.3 99.5 165 2B 7.6 7.2 93 99.0 0.37 0.01 55 6.0 10.0 24.6 1.3 5.7 96.4 65 6.0 10.0 34.7 1.5 5.9 96.3 TABLE 10 IMPURITIES IN TSLP ALKALINE FILTRATE Neutralization Leach Conc.","Temp Na2CO3 NaOH Residue Soluble Filtrate Filtrate Impurities, mg/l No.",": Feed ° C. to ?","pH to ?","pH Solids Mo Mo, g/l Cu Fe As Al P Se Mg Si 157 1B 25 7.0 62% 68% 12.6 6 1 4 2 <1 8.0 63% 77% 14.9 8 1 6 1 <1 8.0 9.0 70% 84% 24.6 110 2 8 2 2 158 1B 25 7.0 52% 45% 8.4 2 1 4 2 <1 8.0 56% 74% 14.8 5 1 4 2 <1 8.0 9.0 58% 87% 23.8 47 2 6 2 1 160 1B 25 6.0 9.0 52% 74% 16.8 2 1 6 2 28 161 1B 25 6.0 9.0 60% 92% 17.0 1 1 4 2 76 162 4A 25 6.0 9.0 70% 91% 18.2 6 3 12 0.4 2 28 163 4A 25 6.0 9.0 78% 93% 19.0 8.0 4.0 10.0 0.3 4.4 20 164 2B 35 9.0 57% 94.4% 33.1 35 10.0 55% 95.6% 35.7 55 9.0 60% 92.0% 30.2 55 10.0 58% 98.8% 33.2 75 9.0 56% 95.1% 36.3 75 10.0 54% 95.4% 35.0 55 6.0 9.0 62% 98.5% 20.8 55 6.0 10.0 58% 99.5% 21.0 75 6.0 9.0 65% 97.1% 27.9 75 6.0 10.0 59% 99.5% 29.8 165 2B 55 6.0 10.0 66% 96.4% 24.6 65 6.0 10.0 67% 96.3% 34.7 neutralization with sodium carbonate to 6.0 pH, followed by neutralization with sodium hydroxide to 10.0 pH.","The cosolubilization of impurities was low, with silicon being the highest at 200 mg/l on a 200 g/l Mo basis.","Solvent extraction of molybdenum from the neutralization solution was ideal.","When using a 10% DTDA organic phase and controlling the pH at 4.0 to 4.5 in the extraction mixers, the transfer of arsenic, phosphorus, and sulfate is minimized.","Silicon transfer was moderate with the subsequent ammonium hydroxide strip solution containing about 21 to 71 mg/l silicon on a 200 g/l Mo basis (50 mg/l silicon was the target maximum).","The data indicate two extraction and two strip stages will recover all of the soluble molybdenum into a 200 g/l molybdenum strip solution."]],"string":"[\n [\n \"Production of pure molybdenum oxide from low grade molybdenite concentrates\",\n \"High purity ammonium dimolybdate or molybdenum oxide is produced by the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.\",\n \"The process entails nearly complete oxidation of the sulfide minerals while optimizing the process chemistry and autoclave conditions to solubilize as little of the molybdenum values as possible.\",\n \"The autoclave discharge 12 is then subjected to a leaching step, either an alkaline leach 50, 400 or ammonium leach 250 process, before or after a liquid/solid separation step 20, 220, 410.\",\n \"The solution is then subjected to (a) filtration 60, 410, solvent extraction 70, 440, crystallization 90, 450, and calcination 120, 480 or (b) filtration 260, 280, crystallization 290, and calcination 320 to produce a product suitable for chemical-grade molybdenum oxide 125, 325, 485.\"\n ],\n [\n \"1-54.\",\n \"(canceled) 55.A method of producing high purity ammonium dimolybdate from low grade molybdenite concentrates comprising: a) forming an aqueous slurry of said low grade molybdenite concentrates; b) oxidizing said slurry in an atmosphere containing free oxygen at a pressure of at least about 50 p.s.i.\",\n \"and at a temperature of at least about 200° C. and thereafter producing a first discharge with greater than about 99% of the molybdenum in said low grade molybdenite concentrates oxidized and greater than about 80% of molybdenum values insoluble; c) filtering said first discharge to produce a first liquid filtrate containing soluble molybdenum values and a first solid filter cake containing the insoluble molybdenum values; d) leaching said first solid filter cake with an alkaline solution to produce a second discharge wherein greater than about 98% of the insoluble molybdenum values are solubilized; e) filtering said second discharge to produce a second liquid filtrate and a second solid filter cake; f) recovering the molybdenum values from said second liquid filtrate by solvent extraction with an organic solvent to produce a first liquor by (1) contacting said second liquid filtrate with said organic solvent to form a two-phase mixture and simultaneously reducing the pH level in said two-phase mixture such that the molybdenum values in said second liquid filtrate are extracted into said organic solvent; and (2) stripping said organic solvent with aqueous ammonia to recover the molybdenum values; g) crystallizing said first liquor containing the extracted molybdenum values to produce crystals and a second liquor; and h) recovering said high purity ammonium dimolybdate from said crystals.\",\n \"56.The method of claim 55, wherein the organic solvent contains a secondary amine.\",\n \"57.The method of claim 56, wherein the organic solvent contains ditridecyl amine.\",\n \"58.The method of claim 55, wherein the reduced pH level in said two-phase mixture ranges from about 4.0 to about 4.5.59.The method of claim 55, wherein the reducing is accomplished by adding hydrated sulfuric acid to said two-phase mixture.\",\n \"60.The method of claim 55, wherein ammonium hydroxide is used during the stripping to recover the molybdenum values.\",\n \"61.The method of claim 55, wherein the pH level during the stripping is maintained at about 9.0.62.A method of producing high purity ammonium dimolybdate from low grade molybdenite concentrates comprising: a) forming an aqueous slurry of said low grade molybdenite concentrates; b) oxidizing said slurry in an atmosphere containing free oxygen at an pressure of at least about 50 p.s.i.\",\n \"and at a temperature of at least about 200° C. and thereafter producing a first discharge with greater than about 99% of the molybdenum in said low grade molybdenite concentrates oxidized and greater than about 80% of molybdenum values insoluble; c) separating and filtering said first discharge to produce a first liquid filtrate containing soluble molybdenum values and a first solid filter cake containing the insoluble molybdenum values; d) leaching said first solid filter cake with an ammoniacal solution to produce a second discharge wherein greater than about 98% of the insoluble molybdenum values are solubilized; e) filtering said second discharge to produce a second liquid filtrate and a second solid filter cake; f) aging said second liquid filtrate; g) crystallizing said second liquid filtrate to produce crystals and a first liquor; and h) recovering said high purity ammonium dimolybdate from said crystals; wherein the method further comprises subjecting said first liquid filtrate produced in step (c) to a solvent extraction process to recover molybdenum values, said solvent extraction process comprising contacting said first liquid filtrate with an organic solvent to form a two-phase mixture and simultaneously reducing the pH level in said two-phase mixture such that the molybdenum values in said first liquid filtrate are extracted into said organic solvent, and stripping said organic solvent to produce a second liquor containing the recovered molybdenum values.\",\n \"63.The method of claim 62, wherein the organic solvent contains a secondary amine.\",\n \"64.The method of claim 63, wherein the organic solvent contains ditridecyl amine.\",\n \"65.The method of claim 62, wherein the reduced pH level in said two-phase mixture ranges from about 4.0 to about 4.5.66.The method of claim 62, wherein the reducing is accomplished by adding hydrated sulfuric acid to said two-phase mixture.\",\n \"67.The method of claim 62, wherein sulfuric acid is used during the stripping to recover the molybdenum values.\",\n \"68.The method of claim 62, wherein the pH level during the stripping is maintained at about less than 3.0.69.The method of claim 62, further comprising recycling a portion of said second liquor back to the aqueous slurry in step (a).\",\n \"70.The method of claim 62, further comprising subjecting said second liquor to a cementation process to recover copper values, said cementation process comprising adding iron to said second liquor and mixing to produce a first solution, filtering said first solution to produce a third liquid filtrate and a third solid filter cake, and then recovering said copper values from said third solid filter cake.\",\n \"71.A method of producing high purity ammonium dimolybdate from low grade molybdenite concentrates comprising: a) forming an aqueous slurry of said low grade molybdenite concentrates; b) oxidizing said slurry in an atmosphere containing free oxygen at an pressure of at least about 50 p.s.i.\",\n \"and at a temperature of at least about 200° C. and thereafter producing a first discharge with greater than about 99% of the molybdenum in said low grade molybdenite concentrates oxidized and greater than about 80% of molybdenum values insoluble; c) separating and filtering said first discharge to produce a first liquid filtrate containing soluble molybdenum values and a first solid filter cake containing the insoluble molybdenum values; d) leaching said first solid filter cake with an ammoniacal solution to produce a second discharge wherein greater than about 98% of the insoluble molybdenum values are solubilized; e) filtering said second discharge to produce a second liquid filtrate and a second solid filter cake; f) aging said second liquid filtrate and adding a reagent selected from the group consisting of iron molybdate, ammonium sulfide, sulfide compounds, and ferric sulfate to said second liquid filtrate; g) crystallizing said second liquid filtrate to produce crystals and a first liquor; and h) recovering said high purity ammonium dimolybdate from said crystals.\",\n \"72.A method of producing high purity ammonium dimolybdate from low grade molybdenite concentrates comprising: a) forming an aqueous slurry of said low grade molybdenite concentrates; b) oxidizing said slurry in an atmosphere containing free oxygen at an pressure of at least about 50 p.s.i.\",\n \"and at a temperature of at least about 200° C. and thereafter producing a first discharge with greater than about 99% of the molybdenum in said low grade molybdenite concentrates oxidized and greater than about 80% of molybdenum values insoluble; c) leaching said first discharge with an alkaline solution to produce a second discharge wherein greater than about 98% of the insoluble molybdenum values are solubilized; d) separating and filtering said second discharge to produce a liquid filtrate containing soluble molybdenum values and a solid filter cake containing the insoluble molybdenum values; e) recovering the molybdenum values from said liquid filtrate by solvent extraction with an organic solvent to produce a first liquor by (1) contacting said liquid filtrate with said organic solvent to form a two-phase mixture and simultaneously reducing the pH level in said two-phase mixture such that the molybdenum values in said liquid filtrate are extracted into said organic solvent; and (2) stripping said organic solvent to recover the molybdenum values; f) crystallizing said first liquor containing the extracted molybdenum values to produce crystals and a second liquor; and g) recovering said high purity ammonium dimolybdate from said crystals.\",\n \"73.The method of claim 72, wherein the organic solvent contains a secondary amine.\",\n \"74.The method of claim 73, wherein the organic solvent contains ditridecyl amine.\",\n \"75.The method of claim 72, wherein the reduced pH level in said two-phase mixture ranges from about 4.0 to about 4.5.76.The method of claim 72, wherein the reducing is accomplished by adding hydrated sulfuric acid to said two-phase mixture.\",\n \"77.The method of claim 72, wherein ammonium hydroxide is used during the stripping to recover the molybdenum values.\",\n \"78.The method of claim 72, wherein the pH level during the stripping is maintained at about 9.0.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Extraction of molybdenum from molybdenite-containing materials by an aqueous process has been the subject of study for over 50 years.\",\n \"In 1952, E. S. Usataya 1 reported on the oxidation of molybdenite in water solutions.\",\n \"He found that in neutral, acidic, or weakly alkaline solutions the decomposition products precipitate on the molybdenite surface and protect the molybdenite from further oxidation.\",\n \"He found that strong bases and strong oxidizing agents impede the formation of the protective layers.\",\n \"Increasing temperature accelerated the oxidation rate in alkaline solutions, but up to 60° C. had no effect in acidic solutions.\",\n \"1 Usataya, E. S., “Oxidation of molybdenite in water solutions,” Zapiski Vsesoyuz Mineral Obshschestva, v 81, 298-303 (1952).\",\n \"A Japanese patent 2 was issued for oxygen pressure oxidation (POX) of molybdenite in 1962.The example in this patent leached a 55.5% Mo, 36.4% S, and 4.4% Cu concentrate at 9% solids at 200° C. and 200 atmospheres oxygen.\",\n \"The molybdic acid precipitate that formed during leaching was dissolved using ammonia for recovery of an ammonium molybdate.\",\n \"2 Sada, Koji, “Extraction of molybdenum,” Japanese patent 15.207('62), assigned to Awamura Mining Co., Ltd.\",\n \"In another process disclosure 3 , alkali hydroxide and alternatively ammonium hydroxide was added continuously to the aqueous solution to neutralize the acid as it formed and maintain the pH at 7-12.Other authors 4 postulated the formation of a molybdenum-iron heteropoly complex that decomposes as the acid concentration increases.\",\n \"It also may be a ferrous complex that decomposes as the soluble iron is oxidized to ferric.\",\n \"3 Hallada, Calvin J., et al., “Conversion of molybdenum disulfide to molybdenum oxide,” German patent 2,045,308 (1971).\",\n \"4 Mel'nikov, B. S. and Shapiro, K. A., “Water-autoclave decomposition of molybdenite raw material,” Protsessy Poluch.\",\n \"Rafinirovaniya Tugoplavkikh Met.\",\n \"(1975) 113-120, 253-260.Early applications of molybdenum solvent extraction utilized tertiary amine to extract the molybdenum solubilized by sodium hydroxide leaching of roasted molybdenite calcines.\",\n \"Secondary amines and quaternary ammonium compounds extract anionic molybdenum using a similar chemistry.\",\n \"Amines also were used for extraction of molybdenum from molybdenite roaster scrubber solutions.\",\n \"Molybdenum is an impurity in many uranium ores.\",\n \"When uranium ores are acid leached, some molybdenum reports to the acid leach solution.\",\n \"The tertiary amines readily available during the 1950's and early 1960's tended to have an amine-molybdenum complex with poor solubility in aliphatic diluents (kerosene).\",\n \"Several Russian researchers worked with acid leaching of oxide ores.\",\n \"In many cases the acidity was sufficient for most of the molybdenum to be in a cationic form.\",\n \"Therefore, the cation exchanger (di, 2, ethylhexyl phosphoric acid (DEHPA)) received much study regarding the recovery of molybdenum from complex acid solutions.\",\n \"Karpacheva et al.\",\n \"5 determined that in acid solutions the molybdenum was not present as the simple molybdenyl cation but, the molybdenum was present as polymeric cations.\",\n \"The co-extraction of iron is a major problem when using DEHPA.\",\n \"The authors noted that in a nitric acid system, the acid concentration needs to equal or exceed 3 molar to prevent significant iron extraction.\",\n \"Other authors 6 reported on the benefit of modifiers in reducing the iron coextraction, e.g.\",\n \"tributyl phosphate, dibutyl butyl phosphonate.\",\n \"5 Karpacheva, S. M. et al., “Extraction of molybdenum and iron (III) by di-2-ethylhexyl hydrogen phosphate,” Russian Journal of Inorganic Chemistry, V 12, 7, p 1014-1016 (1967).\",\n \"6 Chiola, Vincent, “Separation of molybdenum values from tungsten values by solvent extraction,” U.S. Pat.\",\n \"No.\",\n \"3,607,008 (1971).\",\n \"Palant et al.\",\n \"7 made a detailed study of the extraction of molybdenum by DEHPA.\",\n \"The solutions studies were prepared by dissolving MoO 3 in sulfuric acid, hydrochloric acid, or nitric acid solutions.\",\n \"Palant, A.\",\n \"A. et. al.\",\n \"“Extraction of molybdenum (VI) with bis(2-ethylhexyl) hydrogen phosphate from an acidic medium,” Inst.\",\n \"Metall.\",\n \"im.\",\n \"Baikova, Moskow, USSR, Report deposited (1979) pp.\",\n \"1-19.Amine exchange has also received much study during the past 40 to 50 years.\",\n \"The difficulties presented by the poor solubility of the amine-molybdenum complex were addressed by using aromatic diluents.\",\n \"MacInnis et al.\",\n \"8 used tri-n-capryl amine (Alamine 336) with the aromatic diluent #28 9 .\",\n \"The authors discuss amine extraction of a complex sulfate-bearing anion.\",\n \"At pH values of 3 and higher, they determined that the ion exchange type mechanism shown in Equation I below predominates.\",\n \"8 MacInnis, M. B., Kim, T. K., and Laferty, J. M., “The use of solvent extraction for the production and recovery of high-purity ammonium paramolybdate from normal alkali molybdate solution,” First Intl Conf on Chemistry and Uses for Molybdenum, p. 56-58 (1973).\",\n \"9 Aromatic diluent #28 is a solvent from 1960 available from Missouri Solvents & Chemicals.\",\n \"The solvent had a boiling range of 165 to 193° C., a flash point of 122° F., a Kauri butanol value of 73, and was 74% aromatics.\",\n \"in-line-formulae description=\\\"In-line Formulae\\\" end=\\\"lead\\\"?\",\n \"2MoS 2 +6H 2 O+9O 2 →2H 2 MoO 4 ↓+4H 2 SO 4 (I) in-line-formulae description=\\\"In-line Formulae\\\" end=\\\"tail\\\"?\",\n \"At pH values below 3, they postulate that the following occurs along with Equation I. in-line-formulae description=\\\"In-line Formulae\\\" end=\\\"lead\\\"?\",\n \"n (R 3 NH.HSO 4 )+(Mo x O y H z ) n .\",\n \"(SO 4 ) m →(R 3 NH) n .\",\n \"(Mo x O y H z ) n .\",\n \"(HSO4) n .\",\n \"(SO 4 ) m (II) in-line-formulae description=\\\"In-line Formulae\\\" end=\\\"tail\\\"?\",\n \"Equation II infers that (Mo x O y H z ) n .\",\n \"(SO 4 ) m is not ionized and Maclnnis postulates some cation transfer.\",\n \"This postulation is based on the fact that 35 S tagged sulfur was found to transfer both from the organic to the aqueous phase and from the aqueous to the organic phase.\",\n \"Litz found in 1970, that tris, tridecyl amine could be used successfully for molybdenum solvent extraction with an aliphatic diluent.\",\n \"There still was potential for formation of insoluble molybdenum-amine complexes, but the molybdenum-tris tridecyl amine complex's solubility in the diluent was much higher than with other tertiary amines.\",\n \"Tris tridecyl amine in an aliphatic diluent was used in a number of pilot circuits for molybdenum solvent extraction from roaster scrubber solutions and to recover byproduct from uranium leach solutions, but it may never have been used in a commercial circuit.\",\n \"The transfer of sulfate from strongly acid solutions is a problem with using amines.\",\n \"Also, the amines are relatively nonselective and will transfer silicon, phosphorus, and arsenic probably as heteropoly compounds.\",\n \"The coextraction of silicon and subsequent solids precipitation during stripping has been a major problem during other studies.\",\n \"The silicon problem was addressed by filtration of the first stripping stage mixture prior to advancing to the settler.\",\n \"Sulfate transfer was high because the solvent could not be fully loaded with molybdenum, i.e., to avoid diluent-insoluble molybdenum-amine complexes, the sulfate transfer was large.\",\n \"Efficient recovery of chemical-grade ammonium dimolybdate (ADM) requires high purity feed solutions containing 200 to 230 g Mo per liter.\",\n \"Impurities in the solution must be removed to avoid inclusion in the ADM. Impurities, that form hydroxides or sulfides, can be removed by additions or pH-control.\",\n \"Other impurities will build up and unless the mother liquor is bled from the crystallization will report to the ADM.\",\n \"Typical molybdenum solvent extraction systems acidulate the feed solution, if necessary, prior to contact with the extractant in the mixer.\",\n \"Generally this means that the extractant is converted to the bisulfate form by acid in the feed solution and then the desired anion exchanges with the bisulfate.\",\n \"When molybdenum is acidulated there is potential for localized high acid concentrations that can form sulfate-bearing molybdenum species.\",\n \"It is an object of the present invention to provide an integrated process for producing high purity ammonium dimolybdate or molybdenum oxide through a process that includes the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.\",\n \"It is a further object of the present invention to provide an improved molybdenum pressure oxidation process which produces a high purity product at reduced capital and operating costs.\",\n \"It is a further object of the present invention to provide an improved solvent extraction method which rejects sulfate and metallic impurities by extracting the molybdenum in an ionic form that contains no sulfate.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The objects set forth above as well as further and other objects and advantages of the present invention are achieved by the present invention now described in summary fashion and with further examples below in preferred embodiments of the practice of the invention.\",\n \"The present invention provides a process of producing a high purity ammonium dimolybdate or molybdenum oxide through the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.\",\n \"The process entails oxidizing the molybdenite concentrates or intermediates in an autoclave operating at greater than 50 p.s.i.\",\n \"oxygen overpressure, preferably between 80-120 p.s.i., at a temperature greater than 200° C., preferably between 210-220° C. to effect almost complete oxidation of the concentrate while optimizing the process chemistry and autoclave conditions to solubilize as little of the molybdenum values as possible.\",\n \"A method of maximizing the insoluble molybdenum values is disclosed in U.S Patent Application entitled “Autoclave Control Mechanisms for Pressure Oxidation of Molybdenite” which is incorporated by reference herein (and a copy of which is provided at Appendix A hereto).\",\n \"The resulting autoclave discharge has greater than 99% of the molybdenum concentrates oxidized and greater than 80% of the molybdenum values insoluble.\",\n \"The autoclave discharge is then subjected to an alkaline leaching of the POX residue using sodium carbonate and sodium hydroxide.\",\n \"More than 99% of the molybdenum dissolves.\",\n \"The molybdenum in this alkaline solution is recovered readily using a secondary amine solvent, di,tridecyl amine (DTDA).\",\n \"The molybdenum is loaded into the organic phase at 4.0 to 4.5 pH.\",\n \"The molybdenum-loaded organic is stripped with ammonium hydroxide to produce solutions suitable for recovery of chemical-grade ADM and ultimately chemical-grade molybdenum oxide.\",\n \"Alternatively, the autoclave discharge may be subjected to an ammoniacal leaching of the POX residue.\",\n \"More than 99% of the molybdenum dissolves.\",\n \"Most of the cosolubilized impurities are precipitated from the ammoniacal leach solution.\",\n \"The solution is evaporated to crystallize chemical grade ADM. An additional route is provided when the leach solution contains more sulfate than is desirable for crystallization of chemical-grade ADM, producing a product suitable for technical grade ADM and ultimately technical grade molybdenum oxide.\",\n \"Alternatively, the POX residue may be subjected to an alkaline leach with sodium carbonate and sodium hydroxide before a liquid-solid separation step such that all of the molybdenum is soluble and the copper and iron transfer to the solids.\",\n \"More than 98% of the molybdenum is transferred to the filtrate from this neutralization.\",\n \"The molybdenum is readily recoverable using the DTDA solvent extraction process.\",\n \"The ammoniacal strip solutions from the DTDA extraction are suitable for recovery of chemical-grade ADM and ultimately chemical-grade molybdenum oxide.\",\n \"Cementation by scrap iron can be used to readily recover the copper either from the raffinate produced from molybdenum solvent extraction of the POX leach solution or directly from the POX leach solution.\",\n \"Gold and silver values transfer to the final leached solid residue produced by each embodiment and are recyclable to a copper smelter.\",\n \"Most of the rhenium, arsenic, and phosphorus are dissolved regardless of the leaching conditions.\",\n \"Other objects, features and advantages of the invention will be apparent from the following description of preferred embodiments thereof, including illustrative non-limiting examples of the practice of the process.\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention relates to the process of producing ammonium dimolybdate for conversion to pure molybdenum oxide or other pure chemicals from molybdenite concentrates and more particularly to a process of producing chemical grade ammonium dimolybdate for conversion to chemical grade molybdenum oxide through a process that includes the pressure oxidation of low-grade molybdenite concentrates.\",\n \"BACKGROUND OF THE INVENTION Extraction of molybdenum from molybdenite-containing materials by an aqueous process has been the subject of study for over 50 years.\",\n \"In 1952, E. S. Usataya1 reported on the oxidation of molybdenite in water solutions.\",\n \"He found that in neutral, acidic, or weakly alkaline solutions the decomposition products precipitate on the molybdenite surface and protect the molybdenite from further oxidation.\",\n \"He found that strong bases and strong oxidizing agents impede the formation of the protective layers.\",\n \"Increasing temperature accelerated the oxidation rate in alkaline solutions, but up to 60° C. had no effect in acidic solutions.\",\n \"1 Usataya, E. S., “Oxidation of molybdenite in water solutions,” Zapiski Vsesoyuz Mineral Obshschestva, v 81, 298-303 (1952).\",\n \"A Japanese patent2 was issued for oxygen pressure oxidation (POX) of molybdenite in 1962.The example in this patent leached a 55.5% Mo, 36.4% S, and 4.4% Cu concentrate at 9% solids at 200° C. and 200 atmospheres oxygen.\",\n \"The molybdic acid precipitate that formed during leaching was dissolved using ammonia for recovery of an ammonium molybdate.\",\n \"2 Sada, Koji, “Extraction of molybdenum,” Japanese patent 15.207('62), assigned to Awamura Mining Co., Ltd.\",\n \"In another process disclosure3, alkali hydroxide and alternatively ammonium hydroxide was added continuously to the aqueous solution to neutralize the acid as it formed and maintain the pH at 7-12.Other authors4 postulated the formation of a molybdenum-iron heteropoly complex that decomposes as the acid concentration increases.\",\n \"It also may be a ferrous complex that decomposes as the soluble iron is oxidized to ferric.\",\n \"3 Hallada, Calvin J., et al., “Conversion of molybdenum disulfide to molybdenum oxide,” German patent 2,045,308 (1971).\",\n \"4 Mel'nikov, B. S. and Shapiro, K. A., “Water-autoclave decomposition of molybdenite raw material,” Protsessy Poluch.\",\n \"Rafinirovaniya Tugoplavkikh Met.\",\n \"(1975) 113-120, 253-260.Early applications of molybdenum solvent extraction utilized tertiary amine to extract the molybdenum solubilized by sodium hydroxide leaching of roasted molybdenite calcines.\",\n \"Secondary amines and quaternary ammonium compounds extract anionic molybdenum using a similar chemistry.\",\n \"Amines also were used for extraction of molybdenum from molybdenite roaster scrubber solutions.\",\n \"Molybdenum is an impurity in many uranium ores.\",\n \"When uranium ores are acid leached, some molybdenum reports to the acid leach solution.\",\n \"The tertiary amines readily available during the 1950's and early 1960's tended to have an amine-molybdenum complex with poor solubility in aliphatic diluents (kerosene).\",\n \"Several Russian researchers worked with acid leaching of oxide ores.\",\n \"In many cases the acidity was sufficient for most of the molybdenum to be in a cationic form.\",\n \"Therefore, the cation exchanger (di, 2, ethylhexyl phosphoric acid (DEHPA)) received much study regarding the recovery of molybdenum from complex acid solutions.\",\n \"Karpacheva et al.5 determined that in acid solutions the molybdenum was not present as the simple molybdenyl cation but, the molybdenum was present as polymeric cations.\",\n \"The co-extraction of iron is a major problem when using DEHPA.\",\n \"The authors noted that in a nitric acid system, the acid concentration needs to equal or exceed 3 molar to prevent significant iron extraction.\",\n \"Other authors6 reported on the benefit of modifiers in reducing the iron coextraction, e.g.\",\n \"tributyl phosphate, dibutyl butyl phosphonate.\",\n \"5 Karpacheva, S. M. et al., “Extraction of molybdenum and iron (III) by di-2-ethylhexyl hydrogen phosphate,” Russian Journal of Inorganic Chemistry, V 12, 7, p 1014-1016 (1967).\",\n \"6 Chiola, Vincent, “Separation of molybdenum values from tungsten values by solvent extraction,” U.S. Pat.\",\n \"No.\",\n \"3,607,008 (1971).\",\n \"Palant et al.7 made a detailed study of the extraction of molybdenum by DEHPA.\",\n \"The solutions studies were prepared by dissolving MoO3 in sulfuric acid, hydrochloric acid, or nitric acid solutions.\",\n \"Palant, A.\",\n \"A. et. al.\",\n \"“Extraction of molybdenum (VI) with bis(2-ethylhexyl) hydrogen phosphate from an acidic medium,” Inst.\",\n \"Metall.\",\n \"im.\",\n \"Baikova, Moskow, USSR, Report deposited (1979) pp.\",\n \"1-19.Amine exchange has also received much study during the past 40 to 50 years.\",\n \"The difficulties presented by the poor solubility of the amine-molybdenum complex were addressed by using aromatic diluents.\",\n \"MacInnis et al.8 used tri-n-capryl amine (Alamine 336) with the aromatic diluent #289.The authors discuss amine extraction of a complex sulfate-bearing anion.\",\n \"At pH values of 3 and higher, they determined that the ion exchange type mechanism shown in Equation I below predominates.\",\n \"8 MacInnis, M. B., Kim, T. K., and Laferty, J. M., “The use of solvent extraction for the production and recovery of high-purity ammonium paramolybdate from normal alkali molybdate solution,” First Intl Conf on Chemistry and Uses for Molybdenum, p. 56-58 (1973).\",\n \"9 Aromatic diluent #28 is a solvent from 1960 available from Missouri Solvents & Chemicals.\",\n \"The solvent had a boiling range of 165 to 193° C., a flash point of 122° F., a Kauri butanol value of 73, and was 74% aromatics.\",\n \"2MoS2+6H2O+9O2→2H2MoO4↓+4H2SO4 (I) At pH values below 3, they postulate that the following occurs along with Equation I. n(R3NH.HSO4)+(MoxOyHz)n.(SO4)m→(R3NH)n.(MoxOyHz)n.(HSO4)n.(SO4)m (II) Equation II infers that (MoxOyHz)n.(SO4)m is not ionized and Maclnnis postulates some cation transfer.\",\n \"This postulation is based on the fact that 35S tagged sulfur was found to transfer both from the organic to the aqueous phase and from the aqueous to the organic phase.\",\n \"Litz found in 1970, that tris, tridecyl amine could be used successfully for molybdenum solvent extraction with an aliphatic diluent.\",\n \"There still was potential for formation of insoluble molybdenum-amine complexes, but the molybdenum-tris tridecyl amine complex's solubility in the diluent was much higher than with other tertiary amines.\",\n \"Tris tridecyl amine in an aliphatic diluent was used in a number of pilot circuits for molybdenum solvent extraction from roaster scrubber solutions and to recover byproduct from uranium leach solutions, but it may never have been used in a commercial circuit.\",\n \"The transfer of sulfate from strongly acid solutions is a problem with using amines.\",\n \"Also, the amines are relatively nonselective and will transfer silicon, phosphorus, and arsenic probably as heteropoly compounds.\",\n \"The coextraction of silicon and subsequent solids precipitation during stripping has been a major problem during other studies.\",\n \"The silicon problem was addressed by filtration of the first stripping stage mixture prior to advancing to the settler.\",\n \"Sulfate transfer was high because the solvent could not be fully loaded with molybdenum, i.e., to avoid diluent-insoluble molybdenum-amine complexes, the sulfate transfer was large.\",\n \"Efficient recovery of chemical-grade ammonium dimolybdate (ADM) requires high purity feed solutions containing 200 to 230 g Mo per liter.\",\n \"Impurities in the solution must be removed to avoid inclusion in the ADM. Impurities, that form hydroxides or sulfides, can be removed by additions or pH-control.\",\n \"Other impurities will build up and unless the mother liquor is bled from the crystallization will report to the ADM.\",\n \"Typical molybdenum solvent extraction systems acidulate the feed solution, if necessary, prior to contact with the extractant in the mixer.\",\n \"Generally this means that the extractant is converted to the bisulfate form by acid in the feed solution and then the desired anion exchanges with the bisulfate.\",\n \"When molybdenum is acidulated there is potential for localized high acid concentrations that can form sulfate-bearing molybdenum species.\",\n \"It is an object of the present invention to provide an integrated process for producing high purity ammonium dimolybdate or molybdenum oxide through a process that includes the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.\",\n \"It is a further object of the present invention to provide an improved molybdenum pressure oxidation process which produces a high purity product at reduced capital and operating costs.\",\n \"It is a further object of the present invention to provide an improved solvent extraction method which rejects sulfate and metallic impurities by extracting the molybdenum in an ionic form that contains no sulfate.\",\n \"SUMMARY OF THE INVENTION The objects set forth above as well as further and other objects and advantages of the present invention are achieved by the present invention now described in summary fashion and with further examples below in preferred embodiments of the practice of the invention.\",\n \"The present invention provides a process of producing a high purity ammonium dimolybdate or molybdenum oxide through the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.\",\n \"The process entails oxidizing the molybdenite concentrates or intermediates in an autoclave operating at greater than 50 p.s.i.\",\n \"oxygen overpressure, preferably between 80-120 p.s.i., at a temperature greater than 200° C., preferably between 210-220° C. to effect almost complete oxidation of the concentrate while optimizing the process chemistry and autoclave conditions to solubilize as little of the molybdenum values as possible.\",\n \"A method of maximizing the insoluble molybdenum values is disclosed in U.S Patent Application entitled “Autoclave Control Mechanisms for Pressure Oxidation of Molybdenite” which is incorporated by reference herein (and a copy of which is provided at Appendix A hereto).\",\n \"The resulting autoclave discharge has greater than 99% of the molybdenum concentrates oxidized and greater than 80% of the molybdenum values insoluble.\",\n \"The autoclave discharge is then subjected to an alkaline leaching of the POX residue using sodium carbonate and sodium hydroxide.\",\n \"More than 99% of the molybdenum dissolves.\",\n \"The molybdenum in this alkaline solution is recovered readily using a secondary amine solvent, di,tridecyl amine (DTDA).\",\n \"The molybdenum is loaded into the organic phase at 4.0 to 4.5 pH.\",\n \"The molybdenum-loaded organic is stripped with ammonium hydroxide to produce solutions suitable for recovery of chemical-grade ADM and ultimately chemical-grade molybdenum oxide.\",\n \"Alternatively, the autoclave discharge may be subjected to an ammoniacal leaching of the POX residue.\",\n \"More than 99% of the molybdenum dissolves.\",\n \"Most of the cosolubilized impurities are precipitated from the ammoniacal leach solution.\",\n \"The solution is evaporated to crystallize chemical grade ADM. An additional route is provided when the leach solution contains more sulfate than is desirable for crystallization of chemical-grade ADM, producing a product suitable for technical grade ADM and ultimately technical grade molybdenum oxide.\",\n \"Alternatively, the POX residue may be subjected to an alkaline leach with sodium carbonate and sodium hydroxide before a liquid-solid separation step such that all of the molybdenum is soluble and the copper and iron transfer to the solids.\",\n \"More than 98% of the molybdenum is transferred to the filtrate from this neutralization.\",\n \"The molybdenum is readily recoverable using the DTDA solvent extraction process.\",\n \"The ammoniacal strip solutions from the DTDA extraction are suitable for recovery of chemical-grade ADM and ultimately chemical-grade molybdenum oxide.\",\n \"Cementation by scrap iron can be used to readily recover the copper either from the raffinate produced from molybdenum solvent extraction of the POX leach solution or directly from the POX leach solution.\",\n \"Gold and silver values transfer to the final leached solid residue produced by each embodiment and are recyclable to a copper smelter.\",\n \"Most of the rhenium, arsenic, and phosphorus are dissolved regardless of the leaching conditions.\",\n \"Other objects, features and advantages of the invention will be apparent from the following description of preferred embodiments thereof, including illustrative non-limiting examples of the practice of the process.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is a flow diagram of one embodiment of the process of the present invention in which the POX residue is subjected to an alkaline leach; FIG.\",\n \"2 is a flow diagram of another embodiment of the process of the present invention in which the POX residue is subjected to an ammoniacal leach; and FIG.\",\n \"3 is a flow diagram of another embodiment of the process of the present invention in which the POX residue is subjected to an alkaline leach prior to liquid-solid separation.\",\n \"DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS A process of producing a pure ammonium dimolybdate or molybdenum oxide through the pressure oxidation of low-grade molybdenite concentrates is provided in a first embodiment.\",\n \"Referring to FIG.\",\n \"1, the process begins with the pressure oxidation 10 of low grade molybdenite concentrate or intermediate concentrates at greater than 50 p.s.i.\",\n \"oxygen overpressure, preferably between 80-120 p.s.i., at a temperature greater than 200° C., preferably between 210-220° C. while optimizing the process chemistry and autoclave conditions to solubilize as little of the molybdenum values as possible.\",\n \"This can be accomplished by a high degree of oxidation of sulfide sulfur in the autoclave and acceptably high H2SO4 concentrations.\",\n \"Treated water and recycle wash water from the autoclave filter is added to dilute the feed to the autoclave in order to control the temperature in the autoclave.\",\n \"Recycling a portion of the autoclave discharge slurry 12 back to the autoclave increases the acidity and ferric level in the autoclave.\",\n \"The recycled autoclave discharge aqueous accelerates the leaching rate, thus reducing the reactor size, and the additional acid limits the soluble molybdenum to about 15% of the total.\",\n \"Recycling some of the autoclave slurry 12 back to the autoclave also provides seeding material to increase the size of the MoO3 particles to improve filterability.\",\n \"The resulting autoclave discharge has greater than 99% of the molybdenum concentrates oxidized and greater than 80% of the molybdenum values insoluble.\",\n \"The autoclave discharge slurry is then filtered 20 and the filtrate 22 is treated by scrap iron 24 to precipitate the copper.\",\n \"The cementation process 30 begins by adding the iron 24 to the liquid 22 at room temperature.\",\n \"The scrap iron reacts with the filtrate according to the following equations.\",\n \"Fe+2Fe3+→3Fe2+ (III) Fe+Cu2+→Cu+Fe2+ (V) The recovered copper precipitate will be filtered, washed, and sent to a smelter 32.The chemistry of tail stream 34 from iron cementation 30 is then adjusted and molybdenum is precipitated using metallic iron powder 36 in a stirred tank reactor 40.This product 42 is recycled back to the autoclave feed.\",\n \"If the cementation process 30 is not used, then the filtrate 22 advances directly to the molybdenum precipitation process 40.The insoluble molybdenum values 26 in the POX residue are present as hydrated and anhydrous molybdenum oxides and are readily soluble in alkaline solutions.\",\n \"The autoclave discharge filter cake 26 is then repulped in water and a soda ash solution 28 is added to dissolve the molybdenum 50.Sodium carbonate alone dissolves most of the molybdenum at final pH values below 7.0 but a significant amount of iron was also found to dissolve.\",\n \"Sodium hydroxide is then added to increase the pH level to about 9.0 to 10.0, eliminating the soluble iron and producing a leach slurry having low arsenic, phosphorus, and selenium content.\",\n \"The amount of soluble silicon is variable.\",\n \"The discharge 52 from the soda ash/caustic leach 50 is then filtered and washed 60.The filter cake 62 is sent to the smelter for recovery of precious metals.\",\n \"The filtrate 64 is sent to solvent extraction 70.The sodium molybdate solution 64 produced by the alkaline leach 50 is subjected to an amine solvent extraction 70 of the molybdenum.\",\n \"The molybdenum extraction mixers can be operated to produce a two-phase mixture having either the aqueous- or organic-phase continuous.\",\n \"The organic phase contains 10% DTDA.\",\n \"Controlling the acidity in the mixers at 4.0 to 4.5 pH by the direct addition of hydrated sulfuric acid (65% H2SO4 or so) minimizes the transfer of arsenic, phosphorus, selenium, silicon, and sulfate.\",\n \"The molybdenum-loaded organic then is washed with a weak acid solution and/or water, and then is stripped with an ammonium hydroxide solution.\",\n \"Concentrated ammonium hydroxide or anhydrous, gaseous ammonia is added to control the pH in stripping at about 9.0.After stripping, the organic is water-washed to remove entrained strip solution and the aqueous is advanced as make-up water to stripping.\",\n \"The ideal solvent molybdenum extraction rejects sulfate and metallic impurities.\",\n \"The most efficient method of rejecting sulfate is to extract the molybdenum in an ionic form that contains no sulfate.\",\n \"Typical solvent extraction systems acidulate the feed solution, if necessary, prior to contact with the extractant in the mixer.\",\n \"Generally this means that the extractant is converted to the bisulfate form and then the desired anion exchanges with the bisulfate.\",\n \"In addition, when molybdenum-bearing solutions are acidulated there is potential for localized high acid concentrations that can form sulfate-bearing molybdenum specie.\",\n \"The molybdenum ion present in neutral to alkaline solutions is the molybdate ion.\",\n \"On mild acidulation to 4.0 to 4.5 pH, the molybdenum will form large polymolybdate ions that are readily extractable.\",\n \"If the acidulation of the feed solution is done simultaneously in the mixer to produce a two-phase mixture, the extractant quickly exchanges the polymolybdate species before the molybdenum is further acidulated to form a sulfate-bearing specie.\",\n \"Typically the metallic impurities will not form large polyions at the target pH values and, if they do, the polymolybdates will displace them along with any bisulfate ions from the amine.\",\n \"Therefore, small amounts of sulfate and silicon are the only impurities that follow the molybdenum.\",\n \"Raffinate 72 from the solvent extraction is sent to the disposal tank for neutralization and disposal with the other effluents from the plant.\",\n \"The pregnant liquor 74 from the solvent extraction 70 advances to the aging and purification tanks 80 to allow silica to coagulate.\",\n \"Iron molybdate and/or ammonium sulfide 82 may be added to the aging tanks to reduce the amount of trace impurities.\",\n \"The aged and purified solution 84 is then sent to a crystallizer 90 where solid ammonium dimolybdate (ADM) is produced.\",\n \"The crystals are recovered in a centrifuge 100.The vapor 92 from the crystallizer 90 is condensed to recover the ammonia 110 for use in the solvent extraction process 70.The filtrate 102 from the centrifuge is recycled back to the crystallizer 90.As an option, a bleed stream 94 from the crystallizer circuit can be taken to recover rhenium values in a rhenium extraction process 130 and/or the bleed stream 94 can be neutralized with soda ash and steam-stripped to recover and recycle the ammonia.\",\n \"The ammonia-free solution then is returned to the alkaline leach 50.In the rhenium extraction process 130, the rhenium is extracted from the crystallizer mother liquor bleed in ion exchange columns.\",\n \"Stripping is done with ammonia.\",\n \"The strip solution is sent to a crystallizer to recover the rhenium as solid ammonium perrhenate.\",\n \"The crystals 104 from the crystallizer centrifuge 100 are transported to a dryer/calciner 120.The dried/calcined material may then be sent to packaging.\",\n \"The resulting material is suitable for chemical grade ADM or chemical grade molybdenum oxide 125.Table 1 below shows the requirements for chemical grade molybdenum oxide in comparison to technical grade molybdenum oxide.\",\n \"TABLE 1 Technical Grade Chemical Grade MoO3 Spec MoO3 Spec Al, max.\",\n \"ppm — 10 Ca, ppm — 5 Co, ppm — 10 Cr, ppm — 5 Cu, ppm 4500 5 Fe, ppm — 10 K, ppm — 80 Mg, ppm — 5 Mn, ppm — 10 Ni, ppm — 5 P, ppm 200 10 Pb, ppm 300 10 S, ppm 300 — Si, ppm 20,000-40,000 10 Sn, ppm — 10 Ti, ppm — 5 V, ppm — 10 Zn, ppm — 10 Zr, ppm — 10 Referring to FIG.\",\n \"2, in another embodiment of the present invention, the autoclave discharge 12 is centrifuged and washed and then repulped, filtered and washed again 220.The purpose is to obtain a molybdenum-containing filter cake 222 which is low in soluble impurities such as silicon and sulfate.\",\n \"The centrate 224, which contains almost all of the copper and much of the iron from the feed, plus the balance of the molybdenum, is sent to the optional solvent extraction process 230 for molybdenum recovery.\",\n \"The solvent extraction process is essentially the same as described in the first embodiment except the stripping is done with a concentrated sulfuric acid solution rather than an ammonium hydroxide solution.\",\n \"During stripping, the pH is maintained at about less than 3.0.Recovered molybdenum values are recycled back to the autoclave feed.\",\n \"Raffinate 232 from the solvent extraction 230 advances to copper recovery 240, followed by molybdenum precipitation by scrap iron.\",\n \"If the solvent extraction process 230 is not used, then the solution advances directly to copper recovery 240.The filter cake 222 is contacted with ammonium hydroxide solution and anhydrous ammonia 250 rather than the soda ash/caustic leach solution 50 of the first embodiment.\",\n \"The solid molybdic oxide dissolves to form soluble ammonium molybdate.\",\n \"The residue after dissolution is the insoluble portion.\",\n \"The dissolver slurry 252 advances to a continuous filter 260.Some filtrate 262 may be recycled to reduce the solids concentration in the dissolver 250.Both iron molybdate and ammonium sulfide 254 may be added to either the dissolver 250 or the aging tanks 280.These two reagents precipitate trace impurities.\",\n \"The filtrate 262 from the dissolver filter 260 goes to a control tank with a recycle stream back to the dissolver to maintain proper specific gravity and pH.\",\n \"The filtrate from the control tank then goes to the aging and purification tanks 280 to allow silica to coagulate.\",\n \"The aged and purified solution 282 is then sent to a crystallizer 290 where solid ammonium dimolybdate (ADM) is produced.\",\n \"The crystals are recovered in a centrifuge 310.The vapor 292 from the crystallizer 290 is condensed to recover the ammonia 300 for use in the dissolver 250.The majority of the mother-liquor filtrate 312 from the centrifuge 310 is recycled to the crystallizer 290.The crystals from the crystallizer centrifuge 310 are transported to a dryer/calciner 320.The dried/calcined material results in chemical grade ADM or chemical grade molybdenum oxide 325.A part of the mother-liquor filtrate is used as a continuous bleed stream 294 from the crystallizer circuit to control impurities in the mother liquor in the crystallizer 290.This bleed stream 294 is sent back to the dissolver 250 to precipitate the build-up of impurities.\",\n \"If there is a build-up of impurities which can not sufficiently be precipitated in the dissolver/aging circuit, a part of the bleed stream 296 is sent to a second crystallizer 330 to produce an impure ADM.\",\n \"The crystals from the second crystallizer 330 are recovered in a centrifuge 340.The vapor 332 from the second crystallizer 330 is condensed to recover the ammonia 300 for use in the dissolver 250.The crystals from the second crystallizer centrifuge 340 are transported to a dryer/calciner 350.The calcined material results in technical grade ADM or technical grade molybdenum oxide 355, which can be sold or recycled back 252 to the dissolver 250 or the autoclave 10.Referring to FIG.\",\n \"3, in another embodiment of the present invention, the autoclave discharge 12 is subjected to the sodium carbonate/sodium hydroxide leaching process 400 of the first embodiment prior to liquid-solid separation.\",\n \"The leaching discharge 402 is then thickened, centrifuged and washed, repulped, filtered on a continuous pressure filter and rewashed 410.The solids 412 containing the copper and gold are sent to the smelter 422 for recovery.\",\n \"The filtrate 414 containing the molybdenum and rhenium is sent to a rhenium extraction process 430.The rhenium is extracted from the alkaline filtrate in ion exchange columns.\",\n \"Stripping is done with ammonia.\",\n \"The strip solution is sent to a crystallizer to recover the rhenium as solid ammonium perrhenate.\",\n \"The molybdenum-bearing solution 432 from rhenium extraction 430 is pumped to the solvent extraction process 440 previously described in the first embodiment.\",\n \"The raffinate 442 from the solvent extraction 440 is sent to the plant effluent.\",\n \"The pregnant liquor 444 is sent to the crystallization process 450 previously described in the first embodiment producing chemical grade ADM or chemical grade molybdenum oxide 485.EXAMPLES The invention is now further disclosed with reference to the following non-limiting Examples.\",\n \"Example 1 Cementation Process After pressure oxidation of the molybdenite concentrate in the autoclave and the solid-liquid separation of the slurry, the resulting acidic liquid is treated by cementation.\",\n \"The major constituents of the liquid are approximately: Mo, g/l 10-16 Cu, g/l 8-11 Fe, g/l 8-11 H2SO4, g/l 100 The cementation process is conducted at room temperature and begins by adding to the liquid about 1.0 gram Fe per gram Fe present (as Fe3+) in the liquid and about 2.0 gram Fe per gram Cu present in the liquid.\",\n \"The slurry is mixed for 10-15 minutes and then filtered.\",\n \"The solids contain the recovered Cu-values.\",\n \"The filtrate is then treated for the recovery of Mo.\",\n \"The pH of the filtrate is increased to about 1.05-1.2 by the addition of Na2CO3 or NaOH and the temperature of the filtrate is increased to about 40-65° C. An additional 1.5-2.1 gram Fe per gram Mo present in the liquid is then added.\",\n \"The slurry is mixed for 15-30 minutes and filtered.\",\n \"The filtrate should have a clear yellow/greenish color.\",\n \"A recycle stream, which feeds solids or slurry back to the mix-tank is needed to promote the filterability of the Mo-cake.\",\n \"The solids contain the recovered Mo-values.\",\n \"The filtrate can be neutralized with lime and discarded.\",\n \"The mass distribution in % of the elements is shown in Table 2.TABLE 2 Starting Ele- acidic Cu-step, Cu-step, Mo-step, Mo-step, ment liquid Solids Liquid Solids Liquid Notes Mo 100 1-1.5 98.5-99 98.4 0.07-0.24 Cu 100 98.5-99.99 0-1.5 <0.01 0.01 Si 100 15 85 As 100 85 15 Se 100 100 0 (* Re 100 50 50 (* SO4 100 2 6-20 (* P 35 65 (** Mg 30 70 (** Al 10 90 (** Notes: (* = The liquid phase is not analyzed, but based on the synthetic solutions it is assumed that the distribution of the elements are as is indicated.\",\n \"(** = The distribution only indicates the relative distribution between the two solid phases.\",\n \"Example 2 Alkaline Leach A number of tests were performed to attempt optimization of the leaching conditions while minimizing the formation of bicarbonates.\",\n \"The alkaline leaching data are summarized in Table 3.Leaching at pH values below 7.0 using sodium carbonate demonstrated almost complete solubilization of the molybdenum but significant co-solubilization of iron, probably as a carbonate complex, was also present.\",\n \"Leaching with sodium hydroxide did not solubilize much iron.\",\n \"Therefore, tests to optimize leaching cost and efficiency evaluated an initial leach with sodium carbonate to 6 pH, followed by sodium hydroxide to 9 pH or only using sodium hydroxide.\",\n \"Molybdenum extraction in these tests (see Table 3, Leach No.\",\n \"CL-1 to CL-6) exceeded 98%.\",\n \"The leach solutions contained 43 to 79 g/l Mo and silicon was the only impurity of significance.\",\n \"The reagent requirement averaged about 1.1 lb Na2CO3 and 0.7 lb NaOH per lb of molybdenum dissolved.\",\n \"Example 3 Alkaline Leach Solvent Extraction A number of tests were performed to determine the optimum conditions for solvent extracting molybdenum from the alkaline leach solutions.\",\n \"The molybdenum solvent extraction studies used an organic containing 10% di, tridecyl amine, 5% decyl alcohol, and 85% Escaid 110.The results of these studies are summarized in Tables 4A-4C.\",\n \"The initial tests evaluated the effect of temperature in the pH range of 2.0-2.7.Temperature had no significant effect on the molybdenum extraction.\",\n \"The extraction was very efficient with raffinates containing from 1 to 40 mg molybdenum/liter from feed solutions containing 63 to 70 g/l (greater than 99.9% transfer).\",\n \"The distribution coefficients in the first contact, TABLE 3 ALKALINE LEACHING OF POX LEACH RESIDUE MOLYBDENUM EXTRACTION AND LEACH SOLUTION IMPURITIES Dissolution Leach Temp Na2CO3 NaOH Soluble Na2CO3 NaOH Filtrate Filtrate Impurities, mg/l on a 200 g/l Mo basis No.\",\n \": ° C. to ?\",\n \"pH to ?\",\n \"pH Mo g/g Mo g/g Mo Mo, g/l Cu Fe As K P Re Mg Si Se RL-30 50 13 95.6% 0.95 65.3 18 13 37 172 RL-31 50 13 91.9% 0.84 93.6 4 235 RL-32 50 6.8 99.6% 1.31 33.4 24 72 RL-33 50 5.2 99.9% 0.68 31.6 1500 25 468 RL-34 50 3.5 94.5% 0.38 32.2 1500 25 200 RL-35 50 4.3 99.5% 0.39 48.4 1300 8 314 RL-36 50 6.5 98.5% 0.59 76 1040 10 550 RL-37 50 8.05 99.6% 1.00 81.9 <3 7 10 100 RL-38 50 9.54 99.8% 0.99 83.1 <3 5 14 140 RL-39 50 10 95.6% 1.00 74.8 <3 5 5 45 11 27 350 CL-1 50 5.6 9.2 98.0% 1.04 0.72 67.1 12 1500 CL-2 50 5.6 9.4 98.6% 1.04 0.76 58.6 14 2400 CL-3 50 10 98.1% 1.18 74.2 11 890 CL-4 50 9.6 98.4% 1.03 78.8 10 470 CL-5 50 9.0 9.3 98.9% 1.52 0.18 42.9 10 325 CL-6 50 6.9 9.5 99.0% 1.16 0.33 71.1 39 6 124 28 28 TABLE 4A SOLVENT EXTRACTION SUMMARY ALKALINE LEACH SOLUTIONS Impurities Test Temp Mo Content, g/l H2SO4 In strip, mg/l per 200 g/l Mo No.\",\n \"pH ° C. Organic Aqueous g/g Mo Si As P Re SO4 1210 35 Feed 82 110 8 4.8 Loaded Stage 41.5 55 <2 6 1.5 First Contact 27.8 28 0.8 <2 6 0.7 2.5 Raffinate Stage 0.22 <0.001 <2 4 0.05 First Strip Stage, 8.5 pH 3 81 260 25 6.4 Second Strip Stage, 9.5 0.6 7 <55 230 10 1220 Feed 34 2.5 Loaded Stage 45.6 30 4 2.5 First Contact 22.5 0.02 0.41 <2 2.5 Raffinate Stage 0.01 <0.001 <2 First Strip Stage, 8.5 pH 4.1 86 9 107 Second Strip Stage, 9.0 0.3 11 <35 1230 25 Feed 45 76 2 2.5 Loaded Stage 45 47 <2 4 2.5 First Contact 29.6 0.08 <2 <2 2.5 Raffinate Stage 0.01 0.02 <2 4 First Strip Stage, 8.5 pH 9.7 65 215 6 Second Strip Stage, 9.0 1.5 23 70 <15 1410 35 Feed 83 275 6 1700 4.2 Loaded Stage 53.5 61 4.4 First Contact 16.6 0.13 First Strip Stage, 9.0 pH 1.0 183 164 u u 39 Second Strip Stage, 9.0 0.1 6 870 200 <30 370 Re 1420 35 Alkaline Feed 61 10 14 2 10 44 POX Leach Feed 11 80 60 54 26 16 4.4 Alkaline Contact 42.4 25 <2 8 <1 44 20 First POX Contact 7.3 0.33 50 22 28 3 <2 9.1 Strip of Alkaline Contact 4.3 104 31 510 310 200 173 Alkaline Raff Contact 22.7 0.39 <2 6 <1 <1 <1 9.0 Strip of Alkaline Raff Co 0 46 <9 26 9 <9 35600 TABLE 4B SOLVENT EXTRACTION SUMMARY ALKALINE LEACH SOLUTIONS Solvent: 10% DTDA 5% Decyl alcohol 85% Escaid 110 Impurites Test Temp Mo Content, g/l H2SO4 In Strip, mg/l per 200 g/l Mo No.\",\n \"pH ° C. Organic Aqueous g/g Mo Si As P Re 1030 50 Feed 70 56 6 na 12 2.1 Loaded Stage 46.0 1.29 <2 6 0.4 4 1.4 Raffinate Stage 0.06 <0.001 <2 6 0.05 <2 First Strip Stage, 6.5 pH 21 39 <10 <10 5 41 Second Strip Stage, 9.0 p 8 23 313 35 42 <17 1040 50 Feed 70 56 6 na 12 2.6 Loaded Stage 41.6 7.62 <2 6 0.3 4 2.6 Raffinate Stage 0.12 0.016 <2 <2 0.2 <2 First Strip Stage, 6.5 pH 14 43 <9 <9 7 37 Second Strip Stage, 9.0 p 0.5 20 380 <20 25 <20 1070 35 Feed 65 56 6 2.6 Loaded Stage 43.3 0.32 <2 2.8 Raffinate Stage 0.05 0.11 <2 First Strip Stage, 8.5 pH 9.8 63 146 6 Second Strip Stage, 9.5 p 5.6 12 <30 <30 1080 20 Feed 63 56 6 2.7 Loaded Stage 40.9 0.85 2 2.7 Raffinate Stage 0.01 0.003 2 First Strip Stage, 8.5 pH 9.9 72 89 6 Second Strip Stage, 9.5 p 0.1 1.2 <300 <300 1090 35 Feed 70 56 6 12 2.1 Loaded Stage 46.5 0.35 <2 6 2 2.0 Raffinate Stage 0.009 0.001 <2 4 2 First Strip Stage, 8.5 pH 11.8 73 142 5 Second Strip Stage, 9.5 p 0.37 34 200 <5 1100 20 Feed 65 56 6 12 1.95 Loaded Stage 41 3.7 <2 6 2 2.00 Raffinate Stage 0.03 0.04 <2 4 <2 First Strip Stage, 8.5 pH 25.6 32 310 12 Second Strip Stage, 9.5 p 14.4 33 25 <10 TABLE 4C SOLVENT EXTRACTION SUMMARY ALKALINE LEACH SOLUTIONS Impurities Test Temp Mo Content, g/l H2SO4 In strip, mg/l per 200 g/l Mo No.\",\n \"pH ° C. Organic Aqueous g/g Mo Si AS P Re SO4 1110 35 Feed 65 56 6 2.1 Loaded Stage, 120 seco 35.3 13 4 First Strip Stage, 8.5 pH 4.1 35 183 <10 Second Strip Stage, 9.5 2.9 1.3 <300 300 2.1 Loaded Stage, 60 secon 22.5 31 20 4 First Strip Stage, 8.5 pH 3.4 45 187 9 Second Strip Stage, 9.5 2.9 1.5 <250 <250 2.1 Loaded Stage, 30 secon 23.4 30 12 4 First Strip Stage, 8.5 pH 3.7 46 174 9 Second Strip Stage, 9.5 3.3 1.2 <300 <300 2.1 Loaded Stage, 15 secon 16.8 40 20 4 First Strip Stage, 8.5 pH 0 65 197 6 1150 50 Feed 85 110 8 12 3.0 Loaded Stage 41.5 22 <2 6 1 3.1 Raffinate Stage 0.04 0.07 <2 6 First Strip Stage, 8.5 pH 1.3 76 <5 11 16 42 470 Second Strip Stage, 9.5 0.3 2.8 <140 <140 <70 <140 1500 1160 50 Feed 82 110 8 12 3.52 Loaded Stage 39.3 23 <2 6 1 3.66 Raffinate Stage 0.01 0.04 <2 6 First Strip Stage, 8.5 pH 2.1 76 <5 11 13 63 320 Second Strip Stage, 9.5 0.6 4 <100 <100 <50 <100 1700 1170 35 Feed 82 110 8 12 2.5 Loaded Stage 40.7 20 <2 6 8 First Strip Stage, 8.5 pH 1.6 81 197 15 3.7 Second Strip Stage, 9.5 0.3 3.7 750 110 16 freshly stripped organic and feed, were as high as 133 (g/l Mo organic phase÷g/l Mo aqueous phase).\",\n \"There was some transfer of silicon and arsenic.\",\n \"The first strip stages contained up to 310 mg/l silicon and 12 mg/l arsenic when normalized to 200 g/l molybdenum.\",\n \"The effect of contact time on the molybdenum and impurity transfer was also evaluated.\",\n \"As the contact time was reduced from 120 to 15 seconds, the molybdenum transfer was reduced from 80% to 40%.\",\n \"The ratios of silicon and arsenic to molybdenum in subsequent strip solutions did not change, indicating that varying contact time would not vary the amount of impurity transferred.\",\n \"The transfer of sulfate to the strip solutions was also monitored.\",\n \"The pH of the extraction contact was increased.\",\n \"Increasing the pH of the contact reduced the amount of sulfate transferred.\",\n \"The silicon transfer also was reduced in the higher pH contacts.\",\n \"The amount of sulfate transferred to the strip ranged from 320 to 470 mg/l when normalized to 200 g/l molybdenum.\",\n \"The effect of temperature (25, 35, and 50° C.) at pH levels of 2.5 to 3.5 was also evaluated.\",\n \"Within these ranges, temperature and pH appeared to have no effect on the molybdenum transfer.\",\n \"Phase separation was slightly faster at 50° C. The transfer of silicon was lower at 50° C. Sulfate transfer at 50° C. and 3.0 to 3.5 pH was low.\",\n \"Strip solutions contained 320 to 470 mg/l SO4 on a 200 g/l molybdenum basis.\",\n \"A batch contact test was done at the pH found most efficient for molybdenum transfer, 4.2-4.4.The first contact distribution coefficient was 133.The first contact aqueous phase contained 0.13 g/l Mo, indicating 99.8% extraction in the first contact.\",\n \"The transfer of silicon was reduced significantly to 164 mg/l in the strip solution when normalized to 200 g/l Mo.\",\n \"Sulfate transfer also was reduced to 39 mg/l on a 200 g/l Mo basis.\",\n \"Countercurrent Solvent Extraction Two countercurrent extractions were performed and the data from these tests are summarized in Table 5.In both tests a single extraction contact was made.\",\n \"The target pH in the extraction stage was 4.0 to 4.3.The actual pH values of the eight contacts ranged TABLE 5 COUNTERCURRENT SOLVENT EXTRACTION ALKALINE LEACH SOLUTIONS Test No: 1390 Organic: NaOH Leach Aqueous: 10% DTDA 55 g/l Mo 5% Decyl alcohol g/l Fe 85% Escaid 110 g/l H2SO4 mg/l Si Strip Prod Impurities on Loaded Stage 200 g/l Mo basis Contact Mo, g/l Mo, Si, As, P, pH Aqueous Organic g/l mg/l mg/l mg/l Cycle 1 2.3 0.037 217 180 6 Cycle 2 4.0 0.060 224 195 9 Cycle 3 4.3 0.49 221 360 17 Cycle 4 3.8 0.023 226 195 37 Test No: 1400 Organic: NaOH Leach Aqueous: 10% DEHPA 73 g/l Mo 5% Decyl alcohol 275 mg/l Si 85% Escaid 110 3 mg/l P 6 mg/l As 1700 mg/l SO4 Strip Prod Impurities on a Loaded Stage 200 g/l Mo basis Contact Mo, g/l Mo, Si, As, P, SO4, pH Aqueous Organic g/l mg/l mg/l mg/l mg/l Cycle 3.7 14.6 222 240 5 3 65 1 Cycle 4.1 12.9 228 720 5 1 23 2 Cycle 4.2 13.8 226 620 7 3 95 3 Cycle 4.1 14.5 22.7 220 645 5 2 79 4 from 2.3 to 4.3, with only one contact being at a pH lower than 3.7.In this pH range it is easy to add excess acid once the needs for species change were met.\",\n \"Controlling the pH would not be a problem in a continuous circuit.\",\n \"The loaded solvent was water-washed and stripped countercurrently with three stages of ammonium hydroxide.\",\n \"Each test was operated for four cycles.\",\n \"With countercurrent stripping it was possible to produce product strip solutions containing up to 228 g/l Mo.\",\n \"The transfer of sulfate was low, 23 to 95 mg/l on a 200 g/l Mo basis.\",\n \"The strip solutions were well below the target values of phosphorus and arsenic (1-3 and 5-7 mg/l respectively), but contained 240 to 720 mg/l silicon.\",\n \"Example 5 Sulfate Removal from Pressure Oxidation Residue Two washing/re-pulping tests were performed to study removal of sulfate from the autoclave discharge.\",\n \"The results of these tests are shown in Table 6.TABLE 6 Test 1 Test 2 Soluble SO4 per 200 g/l Mo when leached Initial filter cake 250 g/l 250 g/l After 2 displacement washes 12.3 g/l 5.3 g/l Cake after one repulp <4.8 g/l 0.73 g/l After 2 displacement washes <1.4 g/l 0.11 g/l Example 6 Ammoniacal Leaching A series of tests were performed to determine the optimum conditions for ammoniacal leaching of the autoclave residue.\",\n \"The ammoniacal leaching data are summarized in Table 7.The first set of tests evaluated leaching POX residues by adding the wet cake to reagent ammonium hydroxide (nominally 500 g/l NH4OH).\",\n \"The initial TABLE 7 DATA SUMMARY LEACHING OF AUTOCLAVE RESIDUE WITH AMMONIA Test 40 41 42 43 44 45 46 47 Temp 55 55 55 55 55 55 50 50 Pulp Density <43 <38 <79 <79 <79 <79 <82 <79 Stages 1 1 2 2 2 2 2 2 Hours 2 2 2 2 2 2 2 2 Final pH 9.4 9.0 9.0 8.5 9.4 8.8 9.0 9.1 Added iron molybdate no no no no yes yes yes yes Aged overnight no yes yes yes yes yes yes yes Feed, % Mo comp.\",\n \"comp.\",\n \"28.7 27.7 29.3 27.4 25.5 26.4 Residue, % Mo 0.46 0.64 0.16 0.09 0.10 0.25 0.29 0.68 Ammonia soluble, % 99.0 98.4 99.7 99.8 99.8 99.5 99.3 98.3 Filtrate, g/l Mo 31.8 73.7 178 142 158 182 184 181 Impurities per 200 g/l Mo Si 226 190 103 85 56 48 74 82 As <13 33 9 8.5 10 11 11 13 P na 5 6 7 7 8 Cu 19 16 4.5 8.5 11 12 14 14 Fe 132 na 5.6 7 4 3 1 1 K <1 <1 1 1 Mg 2 3 3 2 SO4 380 <330 1300 4240 ammonium hydroxide additions were 1.24 and 1.7 times stoichiometric to dissolve the molybdenum.\",\n \"The mixtures then were heated to 55° C. for two hours and, if necessary, ammonium hydroxide was added to keep the pH above 9.0.The total ammonium hydroxide additions were 1.7 and 3.5 times stoichiometric to dissolve the molybdenum.\",\n \"After the leaching period, the leach slurries were transferred to plastic bottles, sealed, and aged overnight at 50° C. The molybdenum dissolutions were both excellent at 99.7 and 99.8%.\",\n \"The leach filtrates contained 178 and 142 g/l Mo.\",\n \"The soluble impurities on a 200 g/l Mo basis, except for silicon were low at 85-103 mg/l Si, 8.5-9 mg/l As, 2-3 mg/l Mg, <1 mg/l K, 4.5-8.5 mg/l Cu, and 5.6-7 mg/l Fe.\",\n \"The next set evaluated adding iron molybdate and sodium sulfide to the leach slurry prior to the aging period.\",\n \"The initial ammonium hydroxide additions were 1.08 and 1.17 times stoichiometric to dissolve the molybdenum.\",\n \"The mixtures then were heated to 55° C. for two hours and, if necessary, ammonium hydroxide was added to keep the pH above 9.0.The total ammonium hydroxide additions were 2.0 and 1.8 times stoichiometric to dissolve the molybdenum.\",\n \"At 90 minutes iron molybdate was added to the leach slurry and at 105 minutes sodium sulfide was added to the leach slurry.\",\n \"At 120 minutes, the leach slurries were transferred to plastic bottles, sealed, and aged overnight at 50° C. The molybdenum dissolutions were both excellent at 99.8 and 99.5%.\",\n \"The leach filtrates contained 158 and 182 g/l Mo.\",\n \"The soluble impurities on a 200 g/l Mo basis were low at 48-56 mg/l Si, 10-11 mg/l As, 2-3 mg/l Mg, 1 mg/l K, 6-7 mg/l P, 11-12 mg/l Cu, and 3-4 mg/l Fe.\",\n \"The sulfate levels were 380 and less than 330 mg SO4/l.\",\n \"The last set evaluated using anhydrous ammonia to adjust the pH after the initial pulping in ammonium hydroxide solution.\",\n \"The initial ammonium hydroxide additions were 1.65 and 1.72 times stoichiometric to dissolve the molybdenum.\",\n \"The mixtures then were heated to 55° C. for two hours and, if necessary, anhydrous ammonia was added to keep the pH above 9.0.At 90 minutes a small amount of iron molybdate cake was added to the slurry.\",\n \"After the leaching period, the leach slurries were transferred to plastic bottles, sealed, and aged overnight at 55° C. The molybdenum dissolutions were both excellent at 99.3 and 98.3%.\",\n \"The leach filtrate contained 184 and 181 g/l Mo.\",\n \"The soluble impurities on a 200 g/l Mo basis were 74-82 mg/l Si, 11-13 mg/l As, 7-8 mg/l P, 14 mg/l Cu, and 1 mg/l Fe.\",\n \"The sulfate levels in the leach solution were high, 1300 and 4240 mg/l SO4, even though the POX residue had been washed, repulped, refiltered and rewashed.\",\n \"Example 7 Purification of Ammonia Leach Solutions Tests were performed to evaluate purification of leach solutions with high molybdenum concentration, 142-223 g/l Mo.\",\n \"Data are summarized in Table 8.Additives evaluated included ferric sulfate, magnesium sulfate, iron molybdate, and aluminum molybdate.\",\n \"Ferric sulfate and iron molybdate were effective in reducing the silicon by 50 to 70%.\",\n \"Magnesium sulfate had little effect, although the final pH values were below the hydrolysis point of magnesium hydroxide.\",\n \"Aluminum molybdate showed little effect.\",\n \"The addition of iron molybdate successfully reduced the silicon to below the target concentration with each of the solutions.\",\n \"The iron molybdate for the above tests was prepared by mixing solutions of ferric sulfate or ferric chloride with a sodium molybdate solution while controlling the pH at 1.6 to 1.9.Example 8 Total Soda Leach (TSL) Process A series of tests were performed to determine the optimum conditions for neutralizing the autoclave leach slurry prior to solid-liquid separation.\",\n \"The data are summarized in Table 9.This process eliminates one liquid-solid separation step when compared with the alkaline leaching process.\",\n \"Sodium carbonate was added to a fixed pH, 6.0 to 8.0, and then in some cases sodium hydroxide was added to pH 9.0.The data in Table 9 show that the final pH needs to be at least 9.0 for the molybdenum solubility to exceed 90%.\",\n \"The quantities of soluble impurities in the neutralized solution were very low as shown in Table 10.Tests on leach slurries No.\",\n \"164 and 165 focused on determining the lowest cost combination of sodium carbonate and sodium hydroxide that could be used for the neutralization.\",\n \"Because of the formation of bicarbonate, using sodium carbonate is not as cost effective at higher pH values as sodium hydroxide.\",\n \"The leaching-neutralization tests showed that more than 98% of the molybdenum would be soluble after partial TABLE 8 PURIFICATION OF AMMONIUM HYDROXIDE LEACH Test P-41 P-43 P-45 P-42 P-44 P-46 P-47 P-48 P-49 Temp 50 50 50 50 50 50 50 50 50 50 pH 9.0 7.4 7.5 7.3 9.0 7.4 7.4 7.4 8.0 8.5-9.0 9.5 8.5-9.0 8.9 Addition, g/l iron molybdate 28 g 40 g 20 g 10 g wet wet wet wet Ferric sulfate 10 14 Magnesium sulfate hyd 12 16 12 Aluminum molybdate Feed Filtrate Filtrate Filtrate Feed Filtrate Filtrate Filtrate Filtrate Feed Filtrate Feed Filtrate Molybdenum, g/l 142 132 100 155 178 170 134 168 157 184 180 181 180 Impurities per 200 g/l Mo Si 85 42 28 88 103 38 48 114 84 74 40 82 27 As 8 9 12 8 9 7 9 7 8 11 7 13 4 P 6 5 12 6 6 5 10 6 5 7 8 Cu 8 8 12 9 4 >4 >9 7 8 14 9 14 9 Fe 7 9 8 8 6 22 12 7 5 1 1 K 1 2 2 1 1 1 1 1 1 Mg 3 3 32 1400 2 2 15 1400 1300 SO4 1304 4243 TABLE 9 SUMMARY OF TSLP LEACH AND NEUTRALIZATION Leach Conc.\",\n \"Filtrate, g/l Soluble Residue, S % Temp Na2CO3 NaOH Filtrate Residue, % Soluble No.\",\n \": Feed Mo Cu H2SO4 Mo, % AC A Res ° C. to ?\",\n \"pH to ?\",\n \"pH Mo, g/l Mo Cu Mo, % 157 1B 7.1 8.5 65 98.2 25 7.0 12.6 5.6 5.1 68 8.0 14.9 4.1 5.8 77 8.0 9.0 24.6 3.2 4.7 84 158 1B 14.7 6.7 44 99.7 25 7.0 8.4 8.6 4.3 45 8.0 14.8 4.8 4.7 74 8.0 9.0 23.8 2.3 4.6 87 160 1B 20.5 9.0 53 89.3 1.86 1.66 25 6.0 9.0 16.8 4.9 4.1 74 161 1B 5.1 7.1 73 99.3 0.81 0.01 25 6.0 9.0 17.0 2.2 4.9 92 162 4A 9.1 6.5 99.8 0.81 0.10 25 6.0 9.0 18.2 5.2 11.1 91 163 4A 6.1 8.8 99.9 0.10 0.12 25 6.0 9.0 19.0 3.8 11.3 93 164 2B 7.0 3.8 84 99.9 0.25 0.01 35 9.0 33.1 2.6 94.4 35 10.0 35.7 2.0 95.6 55 9.0 30.2 3.7 92.0 55 10.0 33.2 0.6 98.8 75 9.0 36.3 2.3 95.1 75 10.0 35.0 2.0 95.4 55 6.0 9.0 20.8 0.8 98.5 55 6.0 10.0 21.0 0.3 99.5 75 6.0 9.0 27.9 1.3 97.1 75 6.0 10.0 29.8 0.3 99.5 165 2B 7.6 7.2 93 99.0 0.37 0.01 55 6.0 10.0 24.6 1.3 5.7 96.4 65 6.0 10.0 34.7 1.5 5.9 96.3 TABLE 10 IMPURITIES IN TSLP ALKALINE FILTRATE Neutralization Leach Conc.\",\n \"Temp Na2CO3 NaOH Residue Soluble Filtrate Filtrate Impurities, mg/l No.\",\n \": Feed ° C. to ?\",\n \"pH to ?\",\n \"pH Solids Mo Mo, g/l Cu Fe As Al P Se Mg Si 157 1B 25 7.0 62% 68% 12.6 6 1 4 2 <1 8.0 63% 77% 14.9 8 1 6 1 <1 8.0 9.0 70% 84% 24.6 110 2 8 2 2 158 1B 25 7.0 52% 45% 8.4 2 1 4 2 <1 8.0 56% 74% 14.8 5 1 4 2 <1 8.0 9.0 58% 87% 23.8 47 2 6 2 1 160 1B 25 6.0 9.0 52% 74% 16.8 2 1 6 2 28 161 1B 25 6.0 9.0 60% 92% 17.0 1 1 4 2 76 162 4A 25 6.0 9.0 70% 91% 18.2 6 3 12 0.4 2 28 163 4A 25 6.0 9.0 78% 93% 19.0 8.0 4.0 10.0 0.3 4.4 20 164 2B 35 9.0 57% 94.4% 33.1 35 10.0 55% 95.6% 35.7 55 9.0 60% 92.0% 30.2 55 10.0 58% 98.8% 33.2 75 9.0 56% 95.1% 36.3 75 10.0 54% 95.4% 35.0 55 6.0 9.0 62% 98.5% 20.8 55 6.0 10.0 58% 99.5% 21.0 75 6.0 9.0 65% 97.1% 27.9 75 6.0 10.0 59% 99.5% 29.8 165 2B 55 6.0 10.0 66% 96.4% 24.6 65 6.0 10.0 67% 96.3% 34.7 neutralization with sodium carbonate to 6.0 pH, followed by neutralization with sodium hydroxide to 10.0 pH.\",\n \"The cosolubilization of impurities was low, with silicon being the highest at 200 mg/l on a 200 g/l Mo basis.\",\n \"Solvent extraction of molybdenum from the neutralization solution was ideal.\",\n \"When using a 10% DTDA organic phase and controlling the pH at 4.0 to 4.5 in the extraction mixers, the transfer of arsenic, phosphorus, and sulfate is minimized.\",\n \"Silicon transfer was moderate with the subsequent ammonium hydroxide strip solution containing about 21 to 71 mg/l silicon on a 200 g/l Mo basis (50 mg/l silicon was the target maximum).\",\n \"The data indicate two extraction and two strip stages will recover all of the soluble molybdenum into a 200 g/l molybdenum strip solution.\"\n 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"Structure determination of materials using electron microscopy",
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"Magnetic sensor and method for analysing a fluid",
"An apparatus and method for analysing a fluid including means for generating a non-uniform magnetic field in a space and means for measuring the strength of the magnetic field arranged to enable a change in the field strength to be measured when a sampl... | Patent_10475829 |
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"Nucleic acid coding for the cgl1 polypeptide and diagnostic and therapeutic application of said nucleic acid and of the cgl1 polypeptide",
"The invention provides a nucleic acid coding for the CGL1 polypeptide with the SEQ ID No 1 amino acid sequence or also fragments or variants of the CGL1 polypeptide.... | Patent_10475856 |
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"Trinuclear copper-based compound and ligand for nucleic acid scission and anticancer treatment",
"The present invention is related to a novel method for splitting nucleic acids at specific points on a complementary nucleic acid segment using a trinuclear copper-based compound of formula (I).",
"Addit... | Patent_10475860 |
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"Folate mimetics and folate-receptor binding conjugates thereof",
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"The folate mimetic is conjugated to a diagnostic or therapeutic agent to enable selective delivery of the agent to the targeted cell popula... | Patent_10475876 |
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"3-structure of a whole integrin $g(a) v$g(b) extracellular region and uses therefor",
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"Method and apparatus for altering conduction properties in the heart and in adjacent vessels",
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"An ablative catheter is positioned relative to an inter-atria el... | Patent_10475915 |
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"System and method for the measurement of the layer thickness of a multi-layer pipe",
"System for measuring layer thicknesses of a multi-layer pipe by measuring with a detector array (2) the attenuation of an X-ray transmitted though the pipe.",
"According to the invention the detector array (2) compr... | Patent_10475944 |
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"Acetabular prosthesis assembly",
"An acetabular prosthesis assembly (1) comprising an acetabular cup (3) with a generally convex outer surface for engaging acetabular bone and a generally concave inner surface, an insert (10) capable of insertion in the cup (3) for receiving a femoral head component, whe... | Patent_10475948 |
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"Illuminated sign",
"A sign for conveying information, having a plastic body, illuminating means such as light emitting diodes (LEDs) mounted within the body and a reflective coating applied on a surface of the body carrying information to reflect light emitted from the illumination means back into the bo... | Patent_10475953 |
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"Method for improving the operational reliabilty of dosing devices",
"The dispensing of sample liquids is performed, particularly in case of high-throughput screening, by means of automatic dosing devices.",
"Between individual dosing processes, the standstill of the dosing device may lead to an outga... | Patent_10475958 |
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"Methods and compositions for utilizing changes of hybridization signals during approach to equilibrium",
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"Method and device for producing an aqueous acrylamide solutions using a biocatalysts",
"The invention relates to a method and a device for producing an aqueous acrylamide solution by the hydration of acrylnitrile in an aqueous solution in the presence of a biocatalyst."
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"1.Method for produci... | Patent_10475965 |
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"Buffer solution for electroporation and a method comprising the use of the same",
"The invention relates to a buffer solution for suspending animal or human cells and for dissolving biologically active molecules in order to introduce said biologically active molecules into the cells using an electric cur... | Patent_10475967 |
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"Apparatus for performing assess-press-pull massaging",
"The invention concerns an apparatus for massaging the human body, for use in particular in performing an assess/press/pull type massage, comprising a working head (1) open in its lower part and containing means for forming a skin fold.",
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"Hydrogen generation apparatus and method for using same",
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"Method and a device for transferring a leader in a plant for drying of a web-formed material",
"For the purpose of transferring a leader of a web-formed material (1) from a first treatment step to a subsequent second treatment step, the web-formed material (1) is divided by forming a strip (1a) in one ed... | Patent_10476026 |
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"Ballet pointe shoe",
"The invention relates to a custom-fitting, asymmetric ballet shoe with a shell (9) and liner (13) that enables the dancer to stand en pointe and perform the extreme movements required by ballet choreography with minimal discomfort, pain, and injury to the foot caused by the various ... | Patent_10476034 |
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"Apparatus and method for multiple rich media formats video broadcasting",
"Apparatus and method for concurrently converting multiple video and audio formats and sources into different formats.",
"The main elements of the system are: a.",
"A Media Format Matrix (MFM) module, having multiple input ... | Patent_10476081 |
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"Compositions containing plant seed oil",
"The invention relates to palatable compositions comprising a plant seed oil selected from a Ribes fruit, evening primrose or borage plant, a hydrocolloid base, a sweetening agent and an aqueous phase wherein the hydrocolloid base consists of starch and at least o... | Patent_10476103 |
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"Ceramic multilayer substrate manufacturing method and unfired composite multilayer body",
"To produce a green composite laminate 11, a green multilayer collective substrate 13 containing low-temperature sinterable glass ceramic powder as a main ingredient is disposed between first and second shrinkage-re... | Patent_10476155 |
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"Hearing device improvements using modulation techniques",
"There is provided hearing device (140) improvements using modulation techniques adapted to the characteristics of auditory and vestibular hearing.",
"One embodiment provides for extending hearing to the infrasonic range and applying them to a... | Patent_10476158 |
[
[
"Method for distributed multicast routing in connection-oriented networks and network for applying this method",
"A method for multicast routing for a telecommunications network comprising a plurality of communication nodes interconnected by multiple paths, for the establishment of a multicasting connecti... | Patent_10476165 |
[
[
"Signal processing circuit",
"A signal processing circuit comprises a difference stage for receiving an input signal to be processed and a feedback signal taken from an output signal of the circuit.",
"The difference stage generates a difference signal corresponding to the difference between the input... | Patent_10476200 |
[
[
"Process for the production of hydrogen peroxide",
"The invention relates to a process for the production of hydrogen peroxide by the anthraquinone process, comprising a hydrogenation stage, an oxidation stage and an extraction stage.",
"According to the invention, catalytic hydrogenation of anthraqui... | Patent_10476202 |
[
[
"Method and device for evaluating a parameter of a moving object",
"A method for measuring a parameter of a movement of an object, in which a series of successive first time-spaced linear images and a second spaced linear image are taken, (a) a first development of first linear images and (b) a second dev... | Patent_10476207 |
[
[
"Control of weed with a fungal pathogen",
"The present invention provides an isolated biocontrol agent, which is a strain of Pyricularia setariae.",
"The present invention also relates to a biocontrol composition comprising at least one fungal biocontrol agent, which is a strain of Pyricularia setaria... | Patent_10476219 |
[
[
"Fischer tropsch process",
"A process for the conversion of synthesis gas to product comprising liquid hydrocarbons wherein said process comprises contacting synthesis gas at an elevated temperature and pressure with a mixed particulate catalyst comprising a mixture of a particulate Fischer-Tropsch cataly... | Patent_10476231 |
[
[
"Complexes for transferring substances of interest into a cell",
"The present invention concerns new polar compounds, complexes and compositions comprising them, wherein said compound comprises: (i) a polar headgroup spacer, (ii) at least one hydrophobic moiety, and (iii) at least one hydrophilic polymer,... | Patent_10476247 |
[
[
"Compositions for use in treating ige-associated disorders",
"The present invention provides methods of treating IgE-associated disorders and products for use therein.",
"The methods comprise administering to a subject an amount of a first composition comprising an immunogenie antigen and an amount of... | Patent_10476263 |
[
[
"Enzyme and snp marker for disease",
"The present invention relates to an isolated nucleic acid sequence which encodes a novel protein, where the protein is a new member in a family of enzymes with peptidase activity.",
"Also provided is the use of the nucleic acid sequence and/or protein in medicine ... | Patent_10476264 |
[
[
"Application of fluid bed technology in brewing",
"The aim of the invention is firstly to dry the wort necessary for beer production and subsequently to re-dissolve the above for the further processing, in particular, the fermentation.",
"According to the invention, the application of fluid bed techno... | Patent_10476315 |
[
[
"Optical beam delivery configuration",
"An optical beam delivery configuration includes zoom lens means (30) and beam scanning means (44) defining an optical path (20) for a light beam.",
"The beam scanning means is disposed after the zoom lens means in the direction of beam delivery.",
"The zoom ... | Patent_10476323 |
[
[
"Right angle measuring device",
"The invention concerns a device comprising a flexible and non-elastic yarn divided by at least three retaining means fixed to predetermined locations and at least three fixing means adapted to support the retaining means and whereof the diameter of each fixing means is sub... | Patent_10476325 |
[
[
"Novel vaccine",
"The present invention relates to intradermal delivery of influenza vaccines, specific influenza formulations and methods for preparing and using them."
],
[
"1.An intradermal delivery device for the intradermal delivery of a flu vaccine, the device comprising: i a container compr... | Patent_10476331 |
[
[
"Chain wheel",
"A chain wheel for a chain drive, in particular for a control drive of an internal combustion engine, comprising a basic body and a gear rim arranged thereon, the basic body and/or said gear rim being produced at least in certain areas thereof from at least one duroplastic material."
],
... | Patent_10476344 |
[
[
"Method for manufacturing stent-grafts",
"A sewing machine is provided which is capable of sewing reinforcing wire to tubular grafts in order to form stent grafts.",
"A bobbin (which may be seated in a shuttle) carries a bottom thread through the bore of the tubular graft and forms a stitch in combina... | Patent_10476349 |
[
[
"Breast cancer-associated genes and uses thereof",
"The present invention relates to seven isolated breast cancer-associated (BCA) polynucleotides, polypeptides, and variants thereof.",
"The invention also relates to BCA antagonists.",
"The invention also encompasses pharmaceutical compositions co... | Patent_10476362 |
[
[
"Arrangement for mounting two coaxial shaftlines",
"In an assembly with two concentric shafts, one of which projects beyond the other, a seal support sleeve placed between them has a conical shape and bears on a spacer of the external shaft, so that an assembly nut of the forward end is accessible to be u... | Patent_10476372 |
[
[
"Novel therapeutic indication of azithromycin for treatment of non-infective inflamatory diseases",
"The invention relates to the use of 9-deoxo-9-dihydro-9a-methyl-9a-homoerythromycin.",
"A (generic name: azithromycin) for the therapy of neutrophil-dominated non-infective inflammatory diseases, pharm... | Patent_10476377 |
[
[
"Schwann cell specific enhancers and methods of use thereof",
"The present invention provides isolated nucleic acid molecules which exhibit transcriptional enhancement in Schwann cells.",
"The present invention also provides vectors comprising the subject isolated nucleic acid molecules.",
"Method... | Patent_10476389 |
[
[
"Multi-band cellular service over catv network",
"A CATV network is modified with a secondary transmission (CBP) by adding filters to separate modified mobile-communications frequencies.",
"A CATV terminal can be routed to cellular network by way network coupling devices (NCD)."
],
[
"1.A meth... | Patent_10476412 |
[
[
"Thread cutting tap and a method of its manufacture",
"A thread cutting tap having a threaded portion for cutting out a thread in a wall of a workpiece opening into which the threaded portion is to be introduced.",
"The threaded portion is provided with at least two cutting edges extending in a circum... | Patent_10476417 |
[
[
"Network based system design of custom products with live agent support",
"A network based computer implemented system for designing products is disclosed.",
"The system comprises a network server, such as an Internet web server (30) operable to manage interaction of the system with external computers... | Patent_10476423 |
[
[
"Breast cancer detection",
"A training aid to be used in training women or men to carry out examination of their breasts to give early warning of the development of possibly cancerous lumps.",
"The aid is in the form of a life-size replica torso (10), with two breasts made so that they are as alike as... | Patent_10476433 |
[
[
"Direct hydrocarbon fuel cells",
"A solid oxide fuel cell assembly (SOFC) having an array of SOFC's (12) connected in series by thin-film interconnects (22) are deposited onto a porous, insulating support (14), the layer (12) can be deposited in the order of electrode, electrolyte, interconnect and electr... | Patent_10476434 |
[
[
"Native chemical ligation with three or more components",
"The invention provides a method of assembling oligopeptide intermediates in a native chemical ligation reaction that eliminates self-ligation of bi-functional intermediates.",
"An important aspect of the invention is a bi-functional intermedia... | Patent_10476447 |
[
[
"Internet browsing system",
"An apparatus for Internet browsing and other Internet-interactive activities, comprises: a camera focusing on a printed hypertext document for generating video signals representing an image of the printed hypertext document in electronic form: A processor linked to the camera ... | Patent_10476449 |
[
[
"Novel alkaline protease variants and detergents and cleaning agents containing said novel alkaline protease variants",
"Described herein are novel alkaline protease variants derived from subtilisin.",
"These variants have, with respect to the amino acid sequence of Bacillus lentus subtilisin, variati... | Patent_10476463 |
[
[
"Fragrance composition to be distributed by an aerosol generator",
"The present invention relates to fragrance compositions to be distributed by an aerosol generator.",
"Said fragrance composition comprises 30% by weight or more of a component of the odour class A, wherein said components are characte... | Patent_10476479 |
[
[
"UTILIZATION OF RUTINS AND AESCINS IN THE TREATMENT OF OCULAR CIRCULATORY DISTURBANCES",
"The invention relates to the utilization of rutins and aescins in the treatment of ocular circulatory disturbances.",
"The invention also relates to agents with a corresponding active ingredient combination and a... | Patent_10476484 |
[
[
"Dosage form containing a confectionery composition",
"A dosage form comprises an active ingredient and a confectionery composition.",
"In one embodiment the relative standard deviation of the weight of the dosage form is less than 1%, and the dosage form has at least one face.",
"In another embod... | Patent_10476503 |
[
[
"Ball and a method of manufacturing a ball",
"A spherical ball, for projection through the air to deviate from an expected trajectory, has a first hemisphere (12) and a second hemisphere (14).",
"The surfaces (52, 54) of the first hemisphere (12) and the second hemisphere (14) are generally smooth, ex... | Patent_10476512 |
[
[
"Production of nanocrystal beams",
"Apparatus for producing a beam of clustered atoms comprises a nozzle (17) ejecting atomic clusters from a source (10, 11) in a gas stream.",
"The nozzle (17) comprises a passage having an inwardly tapered inlet portion (17a) and an outwardly tapered outlet portion (... | Patent_10476520 |
[
[
"Composite dosage forms",
"A composite dosage form comprises at least one active ingredient, a first portion comprising a first molded material, and a second portion comprising a second material which is compositionally different from the first material.",
"The first and second portions are joined at ... | Patent_10476529 |
[
[
"Device for homogeneous distribution of a fluid in a chamber and uses thereof",
"The invention concerns a device wherein the diffusion chamber (6) comprises, between its upstream end and its downstream end, in the direction of the fluid flow, at least an insert (12) formed by an annular ring having an ext... | Patent_10476535 |
[
[
"Valve arrangement for surgical instruments and valve cage for accomodating the valve arrangement",
"The invention relates to a valve arrangement for surgical instruments, particularly trocars, such as those used for minimally invasive operations, e.g.",
"operations involving laparoscopic techniques."... | Patent_10476546 |
[
[
"Water soluble cellulose etherified derivatives styptic materials",
"The present invention relates to hemostatic materials made of water-soluble cellulose ether derivatives, such as methylcellulose, ethylcellulose, hydroxyethylcellulose, and pharmaceutically acceptable salts of carboxymethylcellulose, esp... | Patent_10476559 |
[
[
"Radiolucent retractor and related components",
"A radiolucent halo-type retractor (20) having a radiolucent curvilinear fixed member (32) connected to a radiolucent curvilinear movable member (34).",
"Each of the fixed and movable members has a radiolucent male coupling (44, 46) projecting outward fr... | Patent_10476596 |
[
[
"Method and system for fast wake-up of oscillators",
"A method and system for fast wakeup of a high-Q oscillator (300) that includes a resonating element (304), preferably a crystal resonator (304), and an amplifier (310).",
"The method comprises connecting the resonating element (304) to a fast wakeu... | Patent_10476618 |
[
[
"Attractants for moths",
"A method of selecting the components of a blend attractive to moth pests, comprising the steps of: (1) measuring the attractiveness of a plurality of single candidate compounds; and (2) selecting for blending those candidate compounds which show statistically significant attracti... | Patent_10476621 |
[
[
"Phospor layers for electroluminescent panels and methods of their manufacture",
"A phosphor layer for an electroluminescent panel comprises a phosphor powder held in a binder.",
"The binder comprises a mixture of two constituents, one of which is a drying oil or a semi-drying oil or a derivative and ... | Patent_10476645 |
[
[
"System and method for minimizing package delivery time",
"An inventive system for delivering packages includes a computer system for determining an optimum route for delivering the package, an electronic tag associated with the package including a first transceiver, and a signaling device.",
"The inv... | Patent_10476685 |
[
[
"Coating composition",
"A composition which is effective in forming on a substrate a flexible coating that has lubricative properties and which is in the form of an aqueous dispersion having dispersed therein: (a) solid particles of a waxy lubricant; and (b) solid resin particles which are capable of coal... | Patent_10476686 |
[
[
"Method and device for determining movement in at least two successive digital images, computer readable storage medium and computer program",
"An apparatus and method for determining the motion in at least two chronologically successive digital images, the digital images contain pixels which are in each ... | Patent_10476690 |
[
[
"Loudspeakers",
"A method of making an acoustic member for a loudspeaker having an operative frequency range and acoustic output which depends on the values of parameters of geometry, bending stiffness, areal mass distribution, damping, tension modulus, compression modulus and shear modulus of the member,... | Patent_10476695 |
[
[
"Gas absorbing material",
"Disclosed is a gas absorbent material having an excellent gas absorptive capacity, especially a gas absorbent material for use as an interior material for buildings, to absorb harmful gas in the room.",
"The gas absorbent material according to the present invention comprises... | Patent_10476730 |
[
[
"Multilayer polymeric films",
"The use of multilayer film structure comprising at least two unfilled layers of polymeric material substantially devoid of opacifying agent and at least two filled layers of polymeric material wherein said filled layers comprise at least 5% by weight of opacifying agent for ... | Patent_10476733 |
[
[
"Methods for assembling protein microarrays",
"The invention provides methods for assembling protein arrays.",
"In one aspect, the methods comprise use of charged or polar array surfaces in the construction of the array.",
"The methods also comprise use of viscous solutions for the deposition of t... | Patent_10476739 |
[
[
"Dynamic generation of voice application information from a web server",
"A server (410) communicates with a client (435) in a client-server architecture to carry out a dialogue with a user.",
"The client comprises a browser (440) that supports a particular mark-up language, such as voiceXML.",
"T... | Patent_10476746 |
[
[
"N-oxide anthranylamide derivatives and their use as medicaments",
"Substituted N-oxidanthranilamide derivatives, their production and use as pharmaceutical agents for treating diseases that are triggered by persistent angiogenesis are described.",
"The compounds according to the invention can be used... | Patent_10476755 |
[
[
"Control device for supercharged engine",
"The invention is intended to provide good emission-cleaning performance by use of a three-way catalyst alone, without the need for a lean NOx catalyst, while ensuring a fuel economy improvement effect of lean burn operation, and to improve fuel economy up to high... | Patent_10476764 |
[
[
"Machines for washing mops and integrated devices having a pulsator and a water removal barrel thereof",
"The invention provides a machine for washing a mop, which includes a housing, a driving mechanism, a controlling mechanism, a pulsator, a water tank, a water removal barrel, a draining mechanism, and ... | Patent_10476781 |
[
[
"Method",
"The present invention relates to a method for determining a mass changing event in a very small amount of a material of interest (eg a chemical or biological material of interest).",
"The mass changing event may be for example specific binding or nucleotide complementation and may be differ... | Patent_10476804 |
[
[
"Accessory for supporting a shading device",
"An accessories (2) for hands-free support of a shading device (1) with respect to a ground surface (3) by a ground stake (5) or objects such as a picnic table (4), railing or the like by a clamp (14).",
"The accessory (2) comprises complementary components... | Patent_10476807 |
[
[
"Method and system for controlling the transmission of media streams",
"A method and system for controlling the transmission of media streams (31, 32, 33, 34, 35) over a connection (6) that employs a plurality of protocol layers (201, 202, 203) is described, where the number of media streams (31, 32, 33, ... | Patent_10476823 |
[
[
"Immune globulin formulations for the treatment and prevention of an orthopoxvirus infection",
"The invention provides an immune globulin having a high titre of antibody to Orthopoxvirus, pharmaceutical compositions comprising the immune globulin and methods for making same.",
"In one embodiment the i... | Patent_10476853 |
[
[
"Peptide-immobilized substrate and method for measuring target protein",
"Disclosed is a peptide-immobilized substrate for measuring a target protein, with which the peptide can have a structure required for being recognized by the target protein, with which the accurate loading amount of the peptide can ... | Patent_10476861 |
[
[
"Method and apparatus for parameter estimation, modulation classification and interference characterization in satellite communication systems",
"A digital signal processing (DSP)-based approach to parameter estimation modulation identification and interference charaterization in connection with a satelli... | Patent_10476869 |
[
[
"Method and apparatus for measuring adjacent satellite interference",
"A system for determining at least one of uplink ASI and downlink ASI in a satellite communication system having at least a first satellite and a second satellite, each having uplink and downlink footprints and being in communication wi... | Patent_10476870 |
[
[
"Hinge assembly",
"A hinge assembly for a foldable working platform, said hinge assembly comprising a hinge body (22), first and second hinge arms (24,26) connected to the hinge body (22), the first hinge arm being rotatable relative to the hinge body about a first axis of rotation (28), the second hinge ... | Patent_10476907 |
[
[
"Trailing beam suspension with alignment adjustment",
"A trailing beam suspension includes a mechanical adjustment mechanism for aligning the axle and wheels of a vehicle.",
"The trailing beam is mounted to the vehicle frame by a pivot bolt passing through a hanger bracket and a conventional bushed co... | Patent_10476914 |
[
[
"FILTER TOW",
"Abstract of the Disclosure The invention relates to a filter tow made from crimped endless cellulose acetate filaments, whereby the filaments form at least two connected fibre bundles with the same filament titre of at least 600 filaments.",
"The filament titres differ from bundle to bu... | Patent_10476920 |
[
[
"Liquid flavor beverage system",
"A beverage apparatus (30) having a plurality of dispensing modules (32) and a housing (34) that defines a chamber (36) containing water or liquide suitable for use in brewing."
],
[
"1.A liquid flavor dispensing system for use with a beverage making apparatus, the... | Patent_10476928 |
[
[
"Windscreen wiper device",
"A windscreen wiper device comprising an elastic, elongated carrier element, as well as an elongated wiper blade of a flexible material, which can be placed in abutment with a windscreen to be wiped, which wiper blade includes opposing longitudinal grooves on its longitudinal si... | Patent_10476929 |
[
[
"Tunnel first safety device",
"In order to increase the safety of tunnels when a fire occurs, the tunnel is divided into sections (A) which are then separated from each other by curtains (1).",
"Each section (A) is provided with at least one fire sensor (2) which closes said curtains (1) in said secti... | Patent_10476930 |
[
[
"Capacitive dynamometer",
"The invention relates to a capacitive dynamometer comprising an elastic body for receiving a mechanical force to be measured, the body having a measuring cavity, which is deformed, when the mechanical force to be measured is applied to the elastic body, the deformations of the i... | Patent_10476932 |
[
[
"Container positioner",
"A container positioner assembly for use in positioning a volume of material in a container relative to a dispensing apparatus.",
"The container positioner assembly includes structures for engagement with the apparatus allowing relative movement of a container attached thereto.... | Patent_10476949 |
[
[
"Multiple sequencible and ligatible structures for genomic analysis",
"High throughput methods and kits for single nucleotide polymorphism (SNP) genotyping are provided.",
"The methods involve utilizing nested PCR amplification reactions which produce sequencible and ligatible structures.",
"An ou... | Patent_10476970 |
[
[
"Company board data processing system and method",
"A company board data processing system and method provides a user interface platform to supply information related to members of boards of companies to enable comparison of board members or boards at board member level.",
"Parameters related to board... | Patent_10476975 |
[
[
"Polypeptide serving as angiogenic marker and dna thereof",
"A marker for neovascularization, vascular disease, inflammatory disease, entoptic neovascular disease, reproductive system disease, central nervous system disease and cancer, the method of detection of the marker and a diagnosis kit of the disea... | Patent_10476985 |
[
[
"Message authentication",
"There is disclosed a technique of providing message authentication in a communication system comprising the steps of: transmitting a first message from a first device to a second device; transmitting a second message from the second device to the first device, the second message... | Patent_10476988 |
[
[
"Shared water reservoir beverage system",
"An apparatus for making and dispensing a plurality of beverages, the dispensing apparatus comprising; a plurality of beverage dispensing modules (32), each beverage module having a container (68) and a brewing assembly (30) for making a respective beverage and de... | Patent_10477002 |
[
[
"Methods and compositions for prevention of angioproliferation",
"The invention provides pharmaceutical compositions comprising Porphyromonas gingivalis protease and hemagglutinin polypeptides that have anti-angiogenic activity and methods for their use."
],
[
"1.A composition for the treatment, p... | Patent_10477007 |
[
[
"Regulating immune response using dendritic cells",
"Disclosed within a method of altering immune responses using dendritic cells.",
"One form of the method is a method of inducing immunological tolerance in an individual, where type 2 dendritic cells are administrated to an individual, and where the ... | Patent_10477012 |
[
[
"Preservation of competent cells at ambient temperatures",
"The present invention relates to techniques for loading cells with desiccation protectants comprising non-metabolizable and non-reducing carbohydrate analogs.",
"Specifically, competent bacterial cells can be preserved using the loading techn... | Patent_10477014 |
[
[
"Material for coating and product coated with the material",
"The invention relates to an iron based material for coating of surfaces which comprises a binding phase in the form of a corrosion dense alloy which contains at least 50% Fe, 12-30% Cr, maximally 5% Mn, 0-40% Ni, preferably 6-20% Ni and 0-7% Mo... | Patent_10477020 |
[
[
"Waste storage device",
"A container adapted for waste storage is provided.",
"The container has a mechanically operated sealing mechanism that allows a user to seal the waste storage film by depressing an actuator."
],
[
"1.A sealing mechanism for a waste storage container, said waste storage... | Patent_10477041 |
[
[
"Methods and compositions for controlling pests",
"The present invention is directed to pest-controlling compositions comprising as active ingredients one or more β-diones, particularly β-diketones and β-triketones, and to the use of these compositions inter alia for preventing, eradicating, destroying, r... | Patent_10477057 |
[
[
"Production of pure molybdenum oxide from low grade molybdenite concentrates",
"High purity ammonium dimolybdate or molybdenum oxide is produced by the pressure oxidation of low grade molybdenite concentrates or molybdenum intermediates.",
"The process entails nearly complete oxidation of the sulfide ... | Patent_10477080 |