We postulate that MBP -induced release of IL-8 by neutrophils may contribute to the pathophysiology of acute asthma and other inflammatory lung diseases .

We postulate that <protein>MBP</protein> -induced release of <protein>IL-8</protein> by <cell_type>neutrophils</cell_type> may contribute to the pathophysiology of acute asthma and other inflammatory lung diseases .


Expression of either the TCL1 oncogene , or transcripts from its homologue MTCP1/c6.1B , in leukaemic and non-leukaemic T cells from ataxia telangiectasia patients .

Expression of either the <dna>TCL1 oncogene</dna> , or transcripts from its homologue <dna>MTCP1/c6.1B</dna> , in leukaemic and <cell_type>non-leukaemic T cells</cell_type> from ataxia telangiectasia patients .


Patients with the recessively inherited disorder ataxia telangiectasia ( A-T ) have a high level of specific chromosome translocations which can be easily observed in peripheral T cells and show a greatly increased predisposition to leukaemia/lymphoma , mainly of T cell origin .

Patients with the recessively inherited disorder ataxia telangiectasia ( A-T ) have a high level of specific <dna>chromosome translocations</dna> which can be easily observed in <cell_type>peripheral T cells</cell_type> and show a greatly increased predisposition to leukaemia/lymphoma , mainly of <cell_type>T cell</cell_type> origin .


Some translocation cells proliferate into a large clone and may develop into T cell prolymphocytic leukaemia ( T-PLL ) .

Some <cell_type>translocation cells</cell_type> proliferate into a large clone and may develop into T cell prolymphocytic leukaemia ( T-PLL ) .


By the time of diagnosis of T-PLL , the clone contains many more genetic changes in the form of additional translocations .


T-PLL is also seen in non-A-T individuals where expression of either TCL1 ( at 14q32 ) or the c6.1B/MTCP1 A1 transcript ( at-Xq28 ) has been demonstrated in just a few instances .

T-PLL is also seen in non-A-T individuals where expression of either <protein>TCL1</protein> ( at <dna>14q32</dna> ) or the <protein>c6.1B/MTCP1 A1 transcript</protein> ( <dna>at-Xq28</dna> ) has been demonstrated in just a few instances .


We show here , that expression of TCL1 occurs in leukaemic T cells from A-T patients with chromosome 14 rearrangements .

We show here , that expression of <protein>TCL1</protein> occurs in <cell_type>leukaemic T cells</cell_type> from A-T patients with <dna>chromosome 14</dna> rearrangements .


Expression of TCL1 also occurs in the preleukaemic clone cells of A-T patients containing the primary translocation alone .

Expression of <protein>TCL1</protein> also occurs in the <cell_type>preleukaemic clone cells</cell_type> of A-T patients containing the primary translocation alone .


Some expression of TCL1 could also be detected in randomly selected A-T patients without large cytogenetic clones and without any evidence of leukaemic change .

Some expression of <protein>TCL1</protein> could also be detected in randomly selected A-T patients without large cytogenetic clones and without any evidence of leukaemic change .


We also show that expression of the B1 transcript from a second gene , MTCP1 , occurred at a relatively high level only in two T-PLL tumours from A-T patients with t ( X ; 14 ) translocations whereas the MTCP1/A1 transcript is much more widely expressed in both tumour and non tumour cells of A-T and non-A-T individuals .

We also show that expression of the <rna>B1 transcript</rna> from a second gene , <dna>MTCP1</dna> , occurred at a relatively high level only in two T-PLL tumours from A-T patients with t ( X ; 14 ) translocations whereas the <rna>MTCP1/A1 transcript</rna> is much more widely expressed in both <cell_type>tumour and non tumour cells</cell_type> of A-T and non-A-T individuals .


[ Correlation of lymphocytic infiltration of tumor tissue with the hormonal and metabolic state in patients with breast cancer ]


Lymphocyte infiltration of tumor was studied vis-a-vis hormone metabolic status , tumor tissue hormone sensitivity and tobacco smoking , in 113 breast cancer patients , aged 25-77 .


On the average , no correlation was established between degree of lymphocyte infiltration in breast tumor and age and menopause onset .


In smoking menopausal patients , lymphocyte infiltration was found to be higher than in non-smokers ( p < 0.05 ) .


There was a direct correlation between the rate of lymphocyte infiltration and the level of progesterone receptors in tumor .

There was a direct correlation between the rate of lymphocyte infiltration and the level of <protein>progesterone receptors</protein> in tumor .


Some subgroups displayed a direct correlation between infiltration and sex-binding globulin , cholesterol , luteinizing hormone in blood , and lean body mass .

Some subgroups displayed a direct correlation between infiltration and <protein>sex-binding globulin</protein> , cholesterol , luteinizing hormone in blood , and lean body mass .


It was matched by an inverse correlation between lymphocyte infiltration and blood-thyroid hormone concentration , urine catecholamines and free cortisol excretion and fat/lean body mass ratio .


Considering the abovesaid as well as the lymphocyte ability to perform the dual function of immunocytes and hormonocytes , it is suggested that the results may be used in both the study of lymphocyte infiltration and research in means of its control .

Considering the abovesaid as well as the lymphocyte ability to perform the dual function of <cell_type>immunocytes</cell_type> and <cell_type>hormonocytes</cell_type> , it is suggested that the results may be used in both the study of lymphocyte infiltration and research in means of its control .


Gene transcription through activation of G-protein-coupled chemoattractant receptors .

Gene transcription through activation of <protein>G-protein-coupled chemoattractant receptors</protein> .


Receptors for leukocyte chemoattractants , including chemokines , are traditionally considered to be responsible for the activation of special leukocyte functions such as chemotaxis , degranulation , and the release of superoxide anions .

Receptors for <protein>leukocyte chemoattractants</protein> , including <protein>chemokines</protein> , are traditionally considered to be responsible for the activation of special leukocyte functions such as chemotaxis , degranulation , and the release of superoxide anions .


Recently , these G-protein-coupled serpentine receptors have been found to transduce signals leading to gene transcription and translation in leukocytes .

Recently , these <protein>G-protein-coupled serpentine receptors</protein> have been found to transduce signals leading to gene transcription and translation in <cell_type>leukocytes</cell_type> .


Transcription factors , such as NF kappa B and AP-1 , are activated upon stimulation of the cells with several chemoattractants at physiologically relevant concentrations .

<protein>Transcription factors</protein> , such as <protein>NF kappa B</protein> and <protein>AP-1</protein> , are activated upon stimulation of the cells with several chemoattractants at physiologically relevant concentrations .


Activation of transcription factors through these receptors involves G-protein coupling and the activation of protein kinases .

Activation of transcription factors through these receptors involves <protein>G-protein</protein> coupling and the activation of <protein>protein kinases</protein> .


The underlying signaling pathways appear to be different from those utilized by TNF-alpha , a better characterized cytokine that induces the transcription of immediate-early genes .

The underlying signaling pathways appear to be different from those utilized by <protein>TNF-alpha</protein> , a better characterized cytokine that induces the transcription of <dna>immediate-early genes</dna> .


Chemoattractants stimulate the expression of several inflammatory cytokines and chemokines , which in turn may activate their respective receptors and initiate an autocrine regulatory mechanism for persistent cytokine and chemokine gene expression .

Chemoattractants stimulate the expression of several <protein>inflammatory cytokines</protein> and <protein>chemokines</protein> , which in turn may activate their respective receptors and initiate an autocrine regulatory mechanism for persistent <protein>cytokine</protein> and <protein>chemokine</protein> gene expression .


Affinity-driven peptide selection of an NFAT inhibitor more selective than cyclosporin A [ see comments ]


The flow of information from calcium-mobilizing receptors to nuclear factor of activated T cells ( NFAT ) -dependent genes is critically dependent on interaction between the phosphatase calcineurin and the transcription factor NFAT .

The flow of information from <protein>calcium-mobilizing receptors</protein> to <dna>nuclear factor of activated T cells ( NFAT ) -dependent genes</dna> is critically dependent on interaction between the <protein>phosphatase</protein> <protein>calcineurin</protein> and the <protein>transcription factor</protein> <protein>NFAT</protein> .


A high-affinity calcineurin -binding peptide was selected from combinatorial peptide libraries based on the calcineurin docking motif of NFAT .

A high-affinity <protein>calcineurin</protein> -binding peptide was selected from combinatorial peptide libraries based on the <protein>calcineurin docking motif</protein> of <protein>NFAT</protein> .


This peptide potently inhibited NFAT activation and NFAT -dependent expression of endogenous cytokine genes in T cells , without affecting the expression of other cytokines that require calcineurin but not NFAT .

This peptide potently inhibited <protein>NFAT</protein> activation and <protein>NFAT</protein> -dependent expression of <dna>endogenous cytokine genes</dna> in <cell_type>T cells</cell_type> , without affecting the expression of other <protein>cytokines</protein> that require <protein>calcineurin</protein> but not <protein>NFAT</protein> .


Substitution of the optimized peptide sequence into the natural calcineurin docking site increased the calcineurin responsiveness of NFAT .

Substitution of the optimized peptide sequence into the natural <protein>calcineurin docking site</protein> increased the <protein>calcineurin</protein> responsiveness of <protein>NFAT</protein> .


Compounds that interfere selectively with the calcineurin -NFAT interaction without affecting calcineurin phosphatase activity may be useful as therapeutic agents that are less toxic than current drugs .

Compounds that interfere selectively with the <protein>calcineurin</protein> <protein>-NFAT</protein> interaction without affecting <protein>calcineurin</protein> <protein>phosphatase</protein> activity may be useful as therapeutic agents that are less toxic than current drugs .


9-cis retinoic acid induces monocyte chemoattractant protein-1 secretion in human monocytic THP-1 cells .

9-cis retinoic acid induces <protein>monocyte chemoattractant protein-1</protein> secretion in <cell_type>human monocytic THP-1 cells</cell_type> .


Monocyte migration and activation are regulated by monocyte chemoattractant protein-1 ( MCP-1 ) .

Monocyte migration and activation are regulated by <protein>monocyte chemoattractant protein-1</protein> ( <protein>MCP-1</protein> ) .


Prior studies have shown MCP-1 expression is modulated by a variety of ligands that act through extracellular receptors .

Prior studies have shown <protein>MCP-1</protein> expression is modulated by a variety of ligands that act through <protein>extracellular receptors</protein> .


In the current study , we show 9-cis retinoic acid ( RA ) , a ligand for the nuclear hormone receptor retinoid X receptor ( RXR ) and retinoic acid receptor ( RAR ) , markedly induces the expression of MCP-1 .

In the current study , we show 9-cis retinoic acid ( RA ) , a ligand for the <protein>nuclear hormone receptor retinoid X receptor</protein> ( <protein>RXR</protein> ) and <protein>retinoic acid receptor</protein> ( <protein>RAR</protein> ) , markedly induces the expression of <protein>MCP-1</protein> .


In human THP-1 monocytic leukemia cells cultured with RA ( 0.05 to 500 nmol/L ) , MCP-1 expression was induced rapidly , significantly , and dose-dependently by as much as 165-fold .

In <cell_line>human THP-1 monocytic leukemia cells</cell_line> cultured with RA ( 0.05 to 500 nmol/L ) , <protein>MCP-1</protein> expression was induced rapidly , significantly , and dose-dependently by as much as 165-fold .


MCP-1 RNA level was also increased in RA-treated cells .

<rna>MCP-1 RNA</rna> level was also increased in RA-treated cells .


Expression of PPARgamma , a heterodimer partner of RXR , is also markedly induced by RA in THP-1 cells .

Expression of <protein>PPARgamma</protein> , a heterodimer partner of <protein>RXR</protein> , is also markedly induced by RA in <cell_type>THP-1 cells</cell_type> .


However , BRL49653 , a PPARgamma ligand , failed to induce MCP-1 secretion either alone or to modify the expression level induced by RA .

However , BRL49653 , a <protein>PPARgamma</protein> ligand , failed to induce <protein>MCP-1</protein> secretion either alone or to modify the expression level induced by RA .


In contrast , BRL49653 significantly increased MCP-1 ( biotinylated MCP-1 ) binding to THP-1 cells , whereas RA had no effect .

In contrast , BRL49653 significantly increased <protein>MCP-1</protein> ( <protein>biotinylated MCP-1</protein> ) binding to <cell_type>THP-1 cells</cell_type> , whereas RA had no effect .


Other peroxisome proliferator activated receptor ( PPAR ) ligands , 15d-PGJ ( 2 ) and troglitazone ( PPARgamma ) , Wy14 , 643 ( PPARalpha ) , and PD195599 ( PPARbeta ) inhibited the induction of MCP-1 by RA .

Other <protein>peroxisome proliferator activated receptor</protein> ( <protein>PPAR</protein> ) ligands , 15d-PGJ ( 2 ) and troglitazone ( <protein>PPARgamma</protein> ) , Wy14 , 643 ( <protein>PPARalpha</protein> ) , and PD195599 ( <protein>PPARbeta</protein> ) inhibited the induction of <protein>MCP-1</protein> by RA .


RA 's effect on MCP-1 expression in human elutriated monocytes were similar to that observed in the THP-1 cells .

RA 's effect on <protein>MCP-1</protein> expression in human elutriated monocytes were similar to that observed in the <cell_line>THP-1 cells</cell_line> .


These studies identify RA as a nuclear signal for MCP-1 induction in undifferentiated human monocytic cells .

These studies identify RA as a nuclear signal for <protein>MCP-1</protein> induction in <cell_type>undifferentiated human monocytic cells</cell_type> .


These studies also suggest monocyte MCP-1 expression induced through RA may modulate cell migration .

These studies also suggest monocyte <protein>MCP-1</protein> expression induced through RA may modulate cell migration .


Classification of IVS1-10T -- > C as a polymorphism of BRCA1 .

Classification of IVS1-10T -- > C as a polymorphism of <dna>BRCA1</dna> .


Mutations inactivating the tumor suppressor gene BRCA1 may be responsible for disease for up to 80 % of familial ovarian cancer cases .

Mutations inactivating the <dna>tumor suppressor gene</dna> <dna>BRCA1</dna> may be responsible for disease for up to 80 % of familial ovarian cancer cases .


In this syndrome , tumorigenesis classically initiates from an inherited mutation in one allele followed by somatic deletion of the normal allele .

In this syndrome , tumorigenesis classically initiates from an inherited mutation in one <dna>allele</dna> followed by somatic deletion of the <dna>normal allele</dna> .


Sequencing of BRCA1 amplified from genomic DNA of lymphocytes and microdissected ovarian tumor cells of a familial ovarian cancer patient revealed three , rare heterozygous DNA variations ( 2418delA , 233G -- > A , and IVS1-10T -- > C ) in both tumor and constitutional ( lymphocyte ) DNA .

Sequencing of <dna>BRCA1</dna> amplified from <dna>genomic DNA</dna> of <cell_type>lymphocytes</cell_type> and <cell_type>microdissected ovarian tumor cells</cell_type> of a familial ovarian cancer patient revealed three , rare heterozygous DNA variations ( 2418delA , 233G -- > A , and IVS1-10T -- > C ) in both <dna>tumor and constitutional ( lymphocyte ) DNA</dna> .


Thus , both copies of BRCA1 were retained in tumor .

Thus , both copies of <dna>BRCA1</dna> were retained in tumor .


Haplotype analysis of the patient and four siblings assigned 2418delA to one copy of BRCA1 and 233G -- > A and IVS1-10T -- > C to the other .

Haplotype analysis of the patient and four siblings assigned 2418delA to one copy of <dna>BRCA1</dna> and 233G -- > A and IVS1-10T -- > C to the other .


The DNA change , 2418delA , is considered a mutation that inactivated one BRCA1 allele because it caused a frameshift and generation of a premature stop codon , resulting in synthesis of a truncated peptide as evidenced by an in vitro protein truncation test .

The DNA change , 2418delA , is considered a mutation that inactivated one <dna>BRCA1</dna> allele because it caused a frameshift and generation of a <dna>premature stop codon</dna> , resulting in synthesis of a truncated peptide as evidenced by an in vitro protein truncation test .


The DNA variation , 233G -- > A , does not result in an amino acid change , and is considered a benign polymorphism .


IVS1-10T -- > C is a unique BRCA1 change that occurs in the last nucleotide of a consensus sequence for a branch site critical for RNA splicing .

IVS1-10T -- > C is a unique <dna>BRCA1</dna> change that occurs in the last nucleotide of a <dna>consensus sequence</dna> for a <dna>branch site</dna> critical for RNA splicing .


Therefore , we investigated whether IVS1-10T -- > C deleteriously affected BRCA1 splicing or expression , and thereby inactivated the other BRCA1 allele .

Therefore , we investigated whether IVS1-10T -- > C deleteriously affected <dna>BRCA1</dna> splicing or expression , and thereby inactivated the other <dna>BRCA1 allele</dna> .


Using the technique of reverse transcription-polymerase chain reaction ( PCR ) with RNA isolated from lymphoid cell lines of the patient and of controls , no evidence was found that IVS1-10TC abnormally disrupted mRNA splicing or caused the absence of BRCA1 mRNA .

Using the technique of reverse transcription-polymerase chain reaction ( PCR ) with RNA isolated from <cell_line>lymphoid cell lines</cell_line> of the patient and of controls , no evidence was found that IVS1-10TC abnormally disrupted <rna>mRNA</rna> splicing or caused the absence of <rna>BRCA1 mRNA</rna> .


Thus , IVS1-10T -- > C is not harmful to BRCA1 function , and is classified a benign polymorphism .

Thus , IVS1-10T -- > C is not harmful to <dna>BRCA1</dna> function , and is classified a benign polymorphism .


Retention of the normal BRCA1 allele in the tumor with the heterozygous germline BRCA1 mutation , 2418delA , indicated that mutational inactivation of both BRCA1 alleles was not required for tumorigenesis .

Retention of the normal <dna>BRCA1 allele</dna> in the tumor with the heterozygous germline <dna>BRCA1</dna> mutation , 2418delA , indicated that mutational inactivation of both <dna>BRCA1 alleles</dna> was not required for tumorigenesis .


It is possible that the normal allele may be functionally inactivated by a nonmutational mechanism .

It is possible that the <dna>normal allele</dna> may be functionally inactivated by a nonmutational mechanism .


Myb-transformed hematopoietic cells as a model for monocyte differentiation into dendritic cells and macrophages .

<cell_type>Myb-transformed hematopoietic cells</cell_type> as a model for monocyte differentiation into <cell_type>dendritic cells</cell_type> and <cell_type>macrophages</cell_type> .


Immune induction is effected through the interaction of antigen-presenting cells with specific receptors on the surface of thymus-derived lymphocytes .

Immune induction is effected through the interaction of <cell_type>antigen-presenting cells</cell_type> with specific <protein>receptors</protein> on the surface of <cell_type>thymus-derived lymphocytes</cell_type> .


Cells most able to ingest , process , and present antigen appear to be related to the mononuclear phagocyte/neutrophil series .

Cells most able to ingest , process , and present antigen appear to be related to the <cell_type>mononuclear phagocyte/neutrophil series</cell_type> .


For example dendritic cells ( DC ) can be found in colonies of GM-CSF-responsive bone marrow cells , and under experimental conditions are routinely expanded as a population in vitro from GM-CSF-responsive progenitor cells .

For example <cell_type>dendritic cells</cell_type> ( <cell_type>DC</cell_type> ) can be found in colonies of <cell_type>GM-CSF-responsive bone marrow cells</cell_type> , and under experimental conditions are routinely expanded as a population in vitro from <cell_type>GM-CSF-responsive progenitor cells</cell_type> .


To address the question of DC lineage and to determine what genes are involved in lineage commitment , we have generated a series of GM-CSF-responsive cell lines that can be induced to differentiate in a homogeneous manner in vitro .

To address the question of <cell_type>DC lineage</cell_type> and to determine what genes are involved in lineage commitment , we have generated a series of <cell_line>GM-CSF-responsive cell lines</cell_line> that can be induced to differentiate in a homogeneous manner in vitro .


The cloned cell lines are derived from 12-day fetal liver and are transformed with a truncated form of c-myb , which lacks the normal autoregulatory sequences .

The <cell_line>cloned cell lines</cell_line> are derived from 12-day fetal liver and are transformed with a truncated form of <dna>c-myb</dna> , which lacks the normal <dna>autoregulatory sequences</dna> .


As far as we know , these myb-transformed hemopoi-etic cells ( MTHC ) differ from normal only in the unregulated expression of myb , a gene whose expression is obligatory for proliferation of hemopoietic cells .

As far as we know , these <cell_line>myb-transformed hemopoi-etic cells</cell_line> ( <cell_line>MTHC</cell_line> ) differ from normal only in the unregulated expression of myb , a gene whose expression is obligatory for proliferation of hemopoietic cells .


MTHC in the presence of TNF-alpha and IL-4 will differentiate into cells that have many of the properties of macrophages .

<cell_line>MTHC</cell_line> in the presence of <protein>TNF-alpha</protein> and <protein>IL-4</protein> will differentiate into cells that have many of the properties of <cell_type>macrophages</cell_type> .


When the same MTHC lines are exposed to TNF-alpha in combination with IFN-gamma , the cells instead become DC .

When the same <cell_line>MTHC lines</cell_line> are exposed to <protein>TNF-alpha</protein> in combination with <protein>IFN-gamma</protein> , the cells instead become <cell_type>DC</cell_type> .


The differentiated DC are potent presenters of antigen in mixed lymphocyte reactions and of soluble antigen to specific T cell lines .

The differentiated <cell_type>DC</cell_type> are potent presenters of antigen in mixed lymphocyte reactions and of soluble antigen to specific <cell_line>T cell lines</cell_line> .


Thus , cells with the properties of both macrophages and DC can be derived from a single type of GM-CSF-responsive progenitor cell .

Thus , cells with the properties of both <cell_type>macrophages</cell_type> and <cell_type>DC</cell_type> can be derived from a single type of <cell_type>GM-CSF-responsive progenitor cell</cell_type> .


We have used this MTHC system to analyze differences in gene expression as the cells mature along the DC and macrophage pathways .

We have used this <cell_line>MTHC system</cell_line> to analyze differences in gene expression as the cells mature along the <cell_type>DC</cell_type> and <cell_type>macrophage pathways</cell_type> .


A distinctive pattern of differentially expressed cDNAs is evident where macrophage-specific cDNAs are homologous to genes encoding cytoskeletal and cell-surface proteins , whereas the DC -specific cDNAs are homologous to signaling , chemokine , and IFN-gamma-inducible genes .

A distinctive pattern of differentially expressed <dna>cDNAs</dna> is evident where <dna>macrophage-specific cDNAs</dna> are homologous to genes encoding <protein>cytoskeletal and cell-surface proteins</protein> , whereas the <cell_type>DC</cell_type> -specific <dna>cDNAs</dna> are homologous to <dna>signaling , chemokine , and IFN-gamma-inducible genes</dna> .


We discuss the utility of MTHC in analyzing the relationships between DC and macrophages , and suggest that DC and macrophages represent extreme phenotypes in a spectrum of antigen handling cells that are somewhat interchangeable , depending on their immediate environment .

We discuss the utility of <cell_line>MTHC</cell_line> in analyzing the relationships between <cell_type>DC</cell_type> and <cell_type>macrophages</cell_type> , and suggest that <cell_type>DC</cell_type> and <cell_type>macrophages</cell_type> represent extreme phenotypes in a spectrum of <cell_type>antigen handling cells</cell_type> that are somewhat interchangeable , depending on their immediate environment .


The Legionella pneumophila rpoS gene is required for growth within Acanthamoeba castellanii .

The Legionella pneumophila <dna>rpoS gene</dna> is required for growth within Acanthamoeba castellanii .


To investigate regulatory networks in Legionella pneumophila , the gene encoding the homolog of the Escherichia coli stress and stationary-phase sigma factor RpoS was identified by complementation of an E. coli rpoS mutation .

To investigate regulatory networks in Legionella pneumophila , the gene encoding the homolog of the <protein>Escherichia coli stress and stationary-phase sigma factor</protein> <protein>RpoS</protein> was identified by complementation of an E. coli <dna>rpoS</dna> mutation .


An open reading frame that is approximately 60 % identical to the E. coli rpoS gene was identified .

An <dna>open reading frame</dna> that is approximately 60 % identical to the <dna>E. coli rpoS gene</dna> was identified .


Western blot analysis showed that the level of L. pneumophila RpoS increased in stationary phase .

Western blot analysis showed that the level of L. pneumophila <protein>RpoS</protein> increased in stationary phase .


An insertion mutation was constructed in the rpoS gene on the chromosome of L. pneumophila , and the ability of this mutant strain to survive various stress conditions was assayed and compared with results for the wild-type strain .

An insertion mutation was constructed in the <dna>rpoS gene</dna> on the chromosome of L. pneumophila , and the ability of this mutant strain to survive various stress conditions was assayed and compared with results for the wild-type strain .


Both the mutant and wild-type strains were more resistant to stress when in stationary phase than when in the logarithmic phase of growth .


This finding indicates that L. pneumophila RpoS is not required for a stationary-phase-dependent resistance to stress .

This finding indicates that L. pneumophila <protein>RpoS</protein> is not required for a stationary-phase-dependent resistance to stress .


Although the mutant strain was able to kill HL-60- and THP-1-derived macrophages , it could not replicate within a protozoan host , Acanthamoeba castellanii .

Although the mutant strain was able to kill <cell_type>HL-60- and THP-1-derived macrophages</cell_type> , it could not replicate within a protozoan host , Acanthamoeba castellanii .


These data suggest that L. pneumophila possesses a growth phase-dependent resistance to stress that is independent of RpoS control and that RpoS likely regulates genes that enable it to survive in the environment within protozoa .

These data suggest that L. pneumophila possesses a growth phase-dependent resistance to stress that is independent of <protein>RpoS</protein> control and that <protein>RpoS</protein> likely regulates genes that enable it to survive in the environment within protozoa .


Our data indicate that the role of rpoS in L. pneumophila is very different from what has previously been reported for E. coli rpoS .

Our data indicate that the role of <dna>rpoS</dna> in L. pneumophila is very different from what has previously been reported for E. coli <dna>rpoS</dna> .


Monoallelic expression of Pax5 : a paradigm for the haploinsufficiency of mammalian Pax genes ?

Monoallelic expression of <protein>Pax5</protein> : a paradigm for the haploinsufficiency of <dna>mammalian Pax genes</dna> ?


It is generally assumed that most mammalian genes are transcribed from both alleles .

It is generally assumed that most <dna>mammalian genes</dna> are transcribed from both <dna>alleles</dna> .


Hence , the diploid state of the genome offers the advantage that a loss-of-function mutation in one allele can be compensated for by the remaining wild-type allele of the same gene .

Hence , the diploid state of the genome offers the advantage that a loss-of-function mutation in one <dna>allele</dna> can be compensated for by the remaining <dna>wild-type allele</dna> of the same gene .


Indeed , the vast majority of human disease syndromes and engineered mutations in the mouse genome are recessive , indicating that recessiveness is the 'default ' state .

Indeed , the vast majority of human disease syndromes and engineered mutations in the <dna>mouse genome</dna> are recessive , indicating that recessiveness is the 'default ' state .


However , a minority of genes are semi-dominant , as heterozygous loss-of-function mutation in these genes leads to phenotypic abnormalities .


This condition , known as haploinsufficiency , has been described for five of the nine mammalian Pax genes , which are associated with mouse developmental mutants and human disease syndromes .

This condition , known as haploinsufficiency , has been described for five of the nine <dna>mammalian Pax genes</dna> , which are associated with mouse developmental mutants and human disease syndromes .


Recently we have reported that the Pax5 gene is subject to allele -specific regulation during B cell development .

Recently we have reported that the <dna>Pax5 gene</dna> is subject to <dna>allele</dna> -specific regulation during B cell development .


Pax5 is predominantly transcribed from only one of its two alleles in early B-lymphoid progenitors and mature B cells , while it transiently switches to a biallelic mode of transcription in pre-B and immature B cells .

<protein>Pax5</protein> is predominantly transcribed from only one of its two <dna>alleles</dna> in <cell_type>early B-lymphoid progenitors</cell_type> and <cell_type>mature B cells</cell_type> , while it transiently switches to a biallelic mode of transcription in <cell_type>pre-B and immature B cells</cell_type> .


As a consequence , B-lymphoid tissues are mosaic with regard to the transcribed allele , and heterozygous mutation of Pax5 therefore results in deletion of B lymphocytes expressing only the mutant allele .

As a consequence , B-lymphoid tissues are mosaic with regard to the transcribed <dna>allele</dna> , and heterozygous mutation of <protein>Pax5</protein> therefore results in deletion of <cell_type>B lymphocytes</cell_type> expressing only the <dna>mutant allele</dna> .


The allele -specific regulation of Pax5 raises the intriguing possibility that monoallelic expression may also be the mechanism causing the haploinsufficiency of other Pax genes .

The <dna>allele</dna> -specific regulation of <protein>Pax5</protein> raises the intriguing possibility that monoallelic expression may also be the mechanism causing the haploinsufficiency of other <dna>Pax genes</dna> .


In this review , we discuss different models accounting for the haploinsufficiency of mammalian Pax genes , provide further evidence in support of the allele -specific regulation of Pax5 and discuss the implication of these findings in the context of the recent literature describing the stochastic and monoallelic activation of other hematopoietic genes .

In this review , we discuss different models accounting for the haploinsufficiency of <dna>mammalian Pax genes</dna> , provide further evidence in support of the <dna>allele</dna> -specific regulation of <protein>Pax5</protein> and discuss the implication of these findings in the context of the recent literature describing the stochastic and monoallelic activation of other <dna>hematopoietic genes</dna> .


Activation of STAT5 by IL-4 relies on Janus kinase function but not on receptor tyrosine phosphorylation , and can contribute to both cell proliferation and gene regulation .

Activation of <protein>STAT5</protein> by <protein>IL-4</protein> relies on <protein>Janus kinase</protein> function but not on receptor tyrosine phosphorylation , and can contribute to both cell proliferation and gene regulation .


We have investigated mechanisms and consequences of STAT5 activation through the human IL-4 receptor ( IL-4R ) .

We have investigated mechanisms and consequences of <protein>STAT5</protein> activation through the <protein>human IL-4 receptor</protein> ( <protein>IL-4R</protein> ) .


By functionally expressing receptor mutants in the murine pro-B cell line Ba/F3 , we could show that phosphorylated tyrosine residues within the IL-4R alpha chain are dispensable for IL-4 -induced STAT5 activity .

By functionally expressing <protein>receptor mutants</protein> in the <cell_line>murine pro-B cell line</cell_line> <cell_line>Ba/F3</cell_line> , we could show that phosphorylated tyrosine residues within the <protein>IL-4R alpha chain</protein> are dispensable for <protein>IL-4</protein> -induced <protein>STAT5</protein> activity .


However , disruption of a membrane-proximal proline-rich sequence motif ( ' box1 ' ) in either subunit of the bipartite IL-4R abolished not only ligand-induced tyrosine phosphorylation of Janus kinases JAK1 and JAK3 , but also IL-4 -triggered activation of STAT5 and concomitant cell proliferation .

However , disruption of a <protein>membrane-proximal proline-rich sequence motif</protein> ( ' <protein>box1</protein> ' ) in either subunit of the <protein>bipartite IL-4R</protein> abolished not only ligand-induced tyrosine phosphorylation of <protein>Janus kinases</protein> <protein>JAK1</protein> and <protein>JAK3</protein> , but also <protein>IL-4</protein> -triggered activation of <protein>STAT5</protein> and concomitant cell proliferation .


A dominant-negative version of STAT5b , but not of STAT5a , interfered with IL-4 -induced DNA synthesis in Ba/F3 cells , suggesting an involvement of STAT5b in the control of cell proliferation through IL-4R .

A <protein>dominant-negative version</protein> of <protein>STAT5b</protein> , but not of <protein>STAT5a</protein> , interfered with <protein>IL-4</protein> -induced DNA synthesis in <cell_line>Ba/F3 cells</cell_line> , suggesting an involvement of <protein>STAT5b</protein> in the control of cell proliferation through <protein>IL-4R</protein> .


Reporter gene experiments finally showed that transcription from promoters of STAT5 target genes can be specifically induced by challenging cells with IL-4 , and that both STAT5a and STAT5b can contribute to IL-4 -triggered transcriptional control .

Reporter gene experiments finally showed that transcription from promoters of <protein>STAT5</protein> target genes can be specifically induced by challenging cells with <protein>IL-4</protein> , and that both <protein>STAT5a</protein> and <protein>STAT5b</protein> can contribute to <protein>IL-4</protein> -triggered transcriptional control .


Inhibition of NF-AT signal transduction events by a dominant-negative form of calcineurin .

Inhibition of <protein>NF-AT</protein> signal transduction events by a dominant-negative form of <protein>calcineurin</protein> .


An inhibitory , `` dominant-negative , '' form of the calcineurin catalytic ( A ) subunit was prepared , which lacks the calmodulin-binding domain , autoinhibitory domain and most of its catalytic core but possesses the regulatory ( B ) subunit binding domain .

An inhibitory , `` dominant-negative , '' form of the <protein>calcineurin catalytic ( A ) subunit</protein> was prepared , which lacks the <protein>calmodulin-binding domain</protein> , <protein>autoinhibitory domain</protein> and most of its catalytic core but possesses the <protein>regulatory ( B ) subunit</protein> binding domain .


When tested for its ability to block calcineurin -dependent signaling in Jurkat cells , expression of this `` B-subunit knock-out '' ( BKO ) construct suppressed reporter gene activity driven by NF-AT , the pivotal promoter element for interleukin ( IL ) -2 gene induction .

When tested for its ability to block <protein>calcineurin</protein> -dependent signaling in Jurkat cells , expression of this <dna>`` B-subunit knock-out '' ( BKO ) construct</dna> suppressed <dna>reporter gene</dna> activity driven by <protein>NF-AT</protein> , the pivotal <dna>promoter element</dna> for <dna>interleukin ( IL ) -2 gene</dna> induction .


Immunoprecipitation of epitope-labeled BKO demonstrated for the formation of a tight complex with endogenous B subunit in Jurkat cells , consistent with an inhibitory mechanism that involves the sequestration of the B subunit .

Immunoprecipitation of <protein>epitope-labeled BKO</protein> demonstrated for the formation of a tight complex with <protein>endogenous B subunit</protein> in <cell_line>Jurkat cells</cell_line> , consistent with an inhibitory mechanism that involves the sequestration of the <protein>B subunit</protein> .


Furthermore , the sharply reduced NF-AT activity produced by co-transfecting BKO could be `` rescued '' by overexpression of transfected B subunit , suggesting that depletion of this subunit was responsible for the inhibition .

Furthermore , the sharply reduced <protein>NF-AT</protein> activity produced by <dna>co-transfecting BKO</dna> could be `` rescued '' by overexpression of transfected <protein>B subunit</protein> , suggesting that depletion of this subunit was responsible for the inhibition .


These data suggest the potential utility of agents that disrupt calcineurin -mediated signal transduction pathways by blocking formation of the catalytically active dimer of calcineurin A and B subunits .

These data suggest the potential utility of agents that disrupt <protein>calcineurin</protein> -mediated signal transduction pathways by blocking formation of the <protein>catalytically active dimer</protein> of <protein>calcineurin A and B subunits</protein> .


Evaluation of monoclonal anti-D reagents using D variant cells .

Evaluation of monoclonal anti-D reagents using <cell_line>D variant cells</cell_line> .


Monoclonal anti-D antibodies submitted to the Third Monoclonal International Workshop were evaluated against a number of D variant cells using standard serological techniques .

<protein>Monoclonal anti-D antibodies</protein> submitted to the Third Monoclonal International Workshop were evaluated against a number of <cell_line>D variant cells</cell_line> using standard serological techniques .


The monoclonal antibodies were able to discriminate between the cells of Categories Va , VI and DFR but not Category III cells .

The <protein>monoclonal antibodies</protein> were able to discriminate between the cells of <cell_line>Categories Va , VI and DFR but not Category III cells</cell_line> .


Cells within each category did not give any aberrant results .


The Rh : 33 cells behaved as normal Rh ( D ) positive cells .

The <cell_line>Rh : 33 cells</cell_line> behaved as <cell_type>normal Rh ( D ) positive cells</cell_type> .


Interleukin 10 induced c-fos expression in human B cells by activation of divergent protein kinases .

<protein>Interleukin 10</protein> induced <dna>c-fos</dna> expression in <cell_type>human B cells</cell_type> by activation of <protein>divergent protein kinases</protein> .


IL-10 is a potent mediator of human B cell growth and plasma cell formation .

<protein>IL-10</protein> is a potent mediator of <cell_type>human B cell</cell_type> growth and plasma cell formation .


However , signal transduction of IL-10 in B cells is poorly understood .

However , signal transduction of <protein>IL-10</protein> in <cell_type>B cells</cell_type> is poorly understood .


In this study the effect of IL-10 on the expression of the protooncogene c-fos was investigated , because Fos plays a potential role in the regulation of B cell proliferation and differentiation .

In this study the effect of <protein>IL-10</protein> on the expression of the <dna>protooncogene c-fos</dna> was investigated , because Fos plays a potential role in the regulation of B cell proliferation and differentiation .


B cells were purified from buffy coat preparations of healthy blood donors by positive selection using an anti CD20 monoclonal antibody and a MiniMACS separation unit .

<cell_type>B cells</cell_type> were purified from buffy coat preparations of healthy blood donors by positive selection using an <protein>anti CD20 monoclonal antibody</protein> and a MiniMACS separation unit .


B cells were prestimulated with SAC for 48 hrs .

<cell_type>B cells</cell_type> were prestimulated with SAC for 48 hrs .


Then , cells were incubated with medium or IL-10 ( 100 ng/ml ) for 10 to 120 min .

Then , cells were incubated with medium or <protein>IL-10</protein> ( 100 ng/ml ) for 10 to 120 min .


RNA was extracted by phenol/chloroform and c-fos expression was analyzed by PCR assisted mRNA assay .

<rna>RNA</rna> was extracted by phenol/chloroform and <dna>c-fos</dna> expression was analyzed by PCR assisted mRNA assay .


A significant 2-4 fold increase of c-fos expression was observed within 30 min of stimulation with IL-10 ( p < 0.01 ) .

A significant 2-4 fold increase of <dna>c-fos</dna> expression was observed within 30 min of stimulation with <protein>IL-10</protein> ( p < 0.01 ) .


After 2 hrs c-fos expression declined to basal levels .

After 2 hrs <dna>c-fos</dna> expression declined to basal levels .


The effect of IL-10 was dose-dependent with a maximum stimulation using 100 ng/ml of IL-10 .

The effect of <protein>IL-10</protein> was dose-dependent with a maximum stimulation using 100 ng/ml of <protein>IL-10</protein> .


The IL-10 effect on c-fos expression was not blocked by polymyxin B .

The <protein>IL-10</protein> effect on <dna>c-fos</dna> expression was not blocked by polymyxin B .


Using the tyrosine kinase inhibitor genistein ( 10 microM ) a complete inhibition of IL-10 induced c-fos expression was observed .

Using the tyrosine kinase inhibitor genistein ( 10 microM ) a complete inhibition of <protein>IL-10</protein> induced <dna>c-fos</dna> expression was observed .


In addition , H-7 ( 10 microM ) , a specific inhibitor of serine/threonine kinases , significantly blocked IL-10 mediated c-fos expression ( p < 0.05 ) .

In addition , H-7 ( 10 microM ) , a specific inhibitor of <protein>serine/threonine kinases</protein> , significantly blocked <protein>IL-10</protein> mediated <dna>c-fos</dna> expression ( p < 0.05 ) .


In conclusion , these data show that IL-10 induces c-fos expression in human B-cells by activation of tyrosine and serine/threonine kinases .

In conclusion , these data show that <protein>IL-10</protein> induces <dna>c-fos</dna> expression in <cell_type>human B-cells</cell_type> by activation of tyrosine and <protein>serine/threonine kinases</protein> .


Since this is the first report on IL-10 induced signal transduction , these data may help to identify the intracellular mechanisms by which IL-10 stimulates human B-cells .

Since this is the first report on <protein>IL-10</protein> induced signal transduction , these data may help to identify the intracellular mechanisms by which <protein>IL-10</protein> stimulates <cell_type>human B-cells</cell_type> .


Immunocytochemical analysis of MNDA in tissue sections and sorted normal bone marrow cells documents expression only in maturing normal and neoplastic myelomonocytic cells and a subset of normal and neoplastic B lymphocytes .

Immunocytochemical analysis of <protein>MNDA</protein> in tissue sections and sorted normal bone marrow cells documents expression only in maturing normal and neoplastic myelomonocytic cells and a subset of normal and neoplastic B lymphocytes .


The human myeloid cell nuclear differentiation antigen ( MNDA ) is a nuclear antigen known to be expressed in mature myelomonocytic cell lines .

The human myeloid cell nuclear differentiation antigen ( <protein>MNDA</protein> ) is a nuclear antigen known to be expressed in mature myelomonocytic cell lines .


An extensive immunocytochemical evaluation of fixed tissues confirmed MNDA expression in normal maturing granulocytes and monocytes and in acute nonlymphocytic leukemias and chronic myelogenous leukemia .

An extensive immunocytochemical evaluation of fixed tissues confirmed <protein>MNDA</protein> expression in <cell_type>normal maturing granulocytes</cell_type> and <cell_type>monocytes</cell_type> and in acute nonlymphocytic leukemias and chronic myelogenous leukemia .


MNDA was not detected in normal tissue histiocytes but was found in activated macrophages and foreign body giant cells associated with inflammation .

<protein>MNDA</protein> was not detected in normal tissue histiocytes but was found in <cell_type>activated macrophages</cell_type> and <cell_type>foreign body giant cells</cell_type> associated with inflammation .


Flow cytometric cell sorting of normal bone marrow established that MNDA is initially expressed in myeloid blast cells .

Flow cytometric cell sorting of normal bone marrow established that <protein>MNDA</protein> is initially expressed in <cell_type>myeloid blast cells</cell_type> .


Examination of lymphoid tissues showed a low level of expression in a population of normal mande B lymphocytes but not in germinal center cells or plasma cells .

Examination of lymphoid tissues showed a low level of expression in a population of normal <cell_line>mande B lymphocytes</cell_line> but not in <cell_type>germinal center cells</cell_type> or <cell_type>plasma cells</cell_type> .


A subset of B cell neoplasms expressing MNDA included hairy cell leukemia , parafollicular ( monocytoid ) B cell lymphoma , mantle cell lymphoma , and small lymphocytic lymphoma .

A subset of <cell_type>B cell neoplasms</cell_type> expressing <protein>MNDA</protein> included hairy cell leukemia , parafollicular ( monocytoid ) B cell lymphoma , mantle cell lymphoma , and small lymphocytic lymphoma .


Cell sorting of normal bone marrow showed MNDA expression in CD20 +/CD10-/CD5- B cells .

Cell sorting of normal bone marrow showed <protein>MNDA</protein> expression in <protein>CD20</protein> +/CD10-/CD5- <cell_type>B cells</cell_type> .


MNDA was not detected in other normal bone marrow or all other nonhematopoietic cells .

<protein>MNDA</protein> was not detected in other normal bone marrow or all other <cell_type>nonhematopoietic cells</cell_type> .


The hematopoietic cell-specific pattern of MNDA expression was elucidated through a comprehensive analysis of normal and neoplastic tissues , and the results provide further evidence of the coexpression of B- and myeloid cell markers in neoplastic B cells and identify a normal B cell population that might be related to the cell of origin of a subset of B cell neoplasms .

The hematopoietic cell-specific pattern of <protein>MNDA</protein> expression was elucidated through a comprehensive analysis of normal and neoplastic tissues , and the results provide further evidence of the coexpression of <protein>B- and myeloid cell markers</protein> in <cell_type>neoplastic B cells</cell_type> and identify a <cell_type>normal B cell population</cell_type> that might be related to the cell of origin of a subset of <cell_type>B cell neoplasms</cell_type> .


Peroxisome proliferator-activated receptor activators target human endothelial cells to inhibit leukocyte -endothelial cell interaction .

<protein>Peroxisome proliferator-activated receptor activators</protein> target <cell_type>human endothelial cells</cell_type> to inhibit <cell_type>leukocyte</cell_type> <cell_type>-endothelial cell</cell_type> interaction .


An early event in acute and chronic inflammation and associated diseases such as atherosclerosis and rheumatoid arthritis is the induced expression of specific adhesion molecules on the surface of endothelial cells ( ECs ) , which subsequently bind leukocytes .

An early event in acute and chronic inflammation and associated diseases such as atherosclerosis and rheumatoid arthritis is the induced expression of specific <protein>adhesion molecules</protein> on the surface of <cell_type>endothelial cells</cell_type> ( <cell_type>ECs</cell_type> ) , which subsequently bind <cell_type>leukocytes</cell_type> .


Peroxisome proliferator-activated receptors ( PPARs ) , members of the nuclear receptor superfamily of transcription factors , are activated by fatty acid metabolites , peroxisome proliferators , and thiazolidinediones and are now recognized as important mediators in the inflammatory response .

<protein>Peroxisome proliferator-activated receptors</protein> ( <protein>PPARs</protein> ) , members of the <protein>nuclear receptor superfamily</protein> of <protein>transcription factors</protein> , are activated by fatty acid metabolites , peroxisome proliferators , and thiazolidinediones and are now recognized as important mediators in the inflammatory response .


Whether PPAR activators influence the inflammatory responses of ECs is unknown .

Whether PPAR activators influence the inflammatory responses of <cell_type>ECs</cell_type> is unknown .


We show that the PPAR activators 15-deoxy-Delta ( 12 , 14 ) -prostaglandin J ( 2 ) ( 15d-PGJ ( 2 ) ) , Wyeth 14643 , ciglitazone , and troglitazone , but not BRL 49653 , partially inhibit the induced expression of vascular cell adhesion molecule-1 ( VCAM-1 ) , as measured by ELISA , and monocyte binding to human aortic endothelial cells ( HAECs ) activated by phorbol 12-myristate 13-acetate ( PMA ) or lipopolysaccharide .

We show that the PPAR activators 15-deoxy-Delta ( 12 , 14 ) -prostaglandin J ( 2 ) ( 15d-PGJ ( 2 ) ) , Wyeth 14643 , ciglitazone , and troglitazone , but not BRL 49653 , partially inhibit the induced expression of <protein>vascular cell adhesion molecule-1</protein> ( <protein>VCAM-1</protein> ) , as measured by ELISA , and monocyte binding to <cell_type>human aortic endothelial cells</cell_type> ( <cell_type>HAECs</cell_type> ) activated by phorbol 12-myristate 13-acetate ( PMA ) or lipopolysaccharide .


The `` natural '' PPAR activator 15d-PGJ ( 2 ) had the greatest potency and was the only tested molecule capable of partially inhibiting the induced expression of E-selectin and neutrophil-like HL60 cell binding to PMA-activated HAECs .

The `` natural '' PPAR activator 15d-PGJ ( 2 ) had the greatest potency and was the only tested molecule capable of partially inhibiting the induced expression of <protein>E-selectin</protein> and neutrophil-like <cell_line>HL60 cell</cell_line> binding to PMA-activated <cell_type>HAECs</cell_type> .


Intracellular adhesion molecule-1 induction by PMA was unaffected by any of the molecules tested .


Both PPAR-alpha and PPAR-gamma mRNAs were detected in HAECs by using reverse transcription-polymerase chain reaction and a ribonuclease protection assay ; however , we have yet to determine which , if any , of the PPARs are mediating this process .

Both <rna>PPAR-alpha and PPAR-gamma mRNAs</rna> were detected in <cell_type>HAECs</cell_type> by using reverse transcription-polymerase chain reaction and a <protein>ribonuclease</protein> protection assay ; however , we have yet to determine which , if any , of the <protein>PPARs</protein> are mediating this process .


These results suggest that certain PPAR activators may help limit chronic inflammation mediated by VCAM-1 and monocytes without affecting acute inflammation mediated by E-selectin and neutrophil binding .

These results suggest that certain PPAR activators may help limit chronic inflammation mediated by <protein>VCAM-1</protein> and <cell_type>monocytes</cell_type> without affecting acute inflammation mediated by <protein>E-selectin</protein> and <cell_type>neutrophil</cell_type> binding .


Dephosphorylation of ZAP-70 and inhibition of T cell activation by activated SHP1 .

Dephosphorylation of <protein>ZAP-70</protein> and inhibition of T cell activation by <protein>activated SHP1</protein> .


Studies with motheaten mice , which lack the SHP1 protein tyrosine phosphatase , indicate that this enzyme plays an important negative role in T cell antigen receptor ( TCR ) signaling .

Studies with motheaten mice , which lack the <protein>SHP1 protein</protein> <protein>tyrosine phosphatase</protein> , indicate that this enzyme plays an important negative role in <protein>T cell antigen receptor</protein> ( <protein>TCR</protein> ) signaling .


The physiological substrates for SHP1 in T lymphocytes , however , have remained unclear or controversial .

The physiological substrates for <protein>SHP1</protein> in <cell_type>T lymphocytes</cell_type> , however , have remained unclear or controversial .


To define these targets for SHP1 we have compared the effects of constitutively active and inactive mutants of SHP1 on TCR signaling .

To define these targets for <protein>SHP1</protein> we have compared the effects of constitutively active and inactive mutants of <protein>SHP1</protein> on <protein>TCR</protein> signaling .


Expression of wild-type SHP1 had a very small effect on the TCR-induced tyrosine phosphorylation of ZAP-70 and Syk , even when SHP1 was overexpressed 20 - 100-fold over endogenous SHP1 .

Expression of <protein>wild-type SHP1</protein> had a very small effect on the TCR-induced tyrosine phosphorylation of <protein>ZAP-70</protein> and <protein>Syk</protein> , even when <protein>SHP1</protein> was overexpressed 20 - 100-fold over <protein>endogenous SHP1</protein> .


Inactive SHP1-D421A and wild-type SHP2 were without effects .

<protein>Inactive SHP1-D421A</protein> and <protein>wild-type SHP2</protein> were without effects .


Constitutively active SHP1-DeltaSH2 had a more pronounced effect on ZAP-70 and Syk , even when expressed at near physiological levels .

<protein>Constitutively active SHP1-DeltaSH2</protein> had a more pronounced effect on <protein>ZAP-70</protein> and <protein>Syk</protein> , even when expressed at near physiological levels .


SHP1-DeltaSH2 also inhibited events downstream of ZAP-70 and Syk , such as activation of the mitogen-activated protein kinase Erk2 and the transcriptional activation of the interleukin-2 gene .

<protein>SHP1-DeltaSH2</protein> also inhibited events downstream of <protein>ZAP-70</protein> and <protein>Syk</protein> , such as activation of the <protein>mitogen-activated protein kinase</protein> <protein>Erk2</protein> and the transcriptional activation of the <dna>interleukin-2 gene</dna> .


In contrast , a constitutively active SHP2-DeltaSH2 had no statistically significant effect ( although it caused a slight augmentation in some individual experiments ) .

In contrast , a constitutively active <protein>SHP2-DeltaSH2</protein> had no statistically significant effect ( although it caused a slight augmentation in some individual experiments ) .


of the constructs influenced the anti-CD3-induced tyrosine phosphorylation of the TCR zeta-chain or phospholipase Cgamma1 , indicating that Src family kinase function was intact .

of the constructs influenced the anti-CD3-induced tyrosine phosphorylation of the <protein>TCR zeta-chain</protein> or <protein>phospholipase Cgamma1</protein> , indicating that <protein>Src family kinase</protein> function was intact .


Taken together , our findings support the notion that ZAP-70 and Syk can be direct substrates for SHP1 in intact cells .

Taken together , our findings support the notion that <protein>ZAP-70</protein> and <protein>Syk</protein> can be direct substrates for <protein>SHP1</protein> in intact cells .


However , the two SH2 domains of SHP1 did not facilitate its recognition of ZAP-70 and Syk as substrates in intact cells .

However , the two <protein>SH2 domains</protein> of <protein>SHP1</protein> did not facilitate its recognition of <protein>ZAP-70</protein> and <protein>Syk</protein> as substrates in intact cells .


Therefore , we suggest that SHP1 is not actively recruited to inhibit TCR signaling induced by ligation of this receptor alone .

Therefore , we suggest that <protein>SHP1</protein> is not actively recruited to inhibit <protein>TCR</protein> signaling induced by ligation of this receptor alone .


Instead , we propose that ligation of a distinct inhibitory receptor leads to the recruitment of SHP1 via its SH2 domains , activation of SHP1 and subsequently inhibition of TCR signals if the inhibitory receptor is juxtaposed to the TCR .

Instead , we propose that ligation of a distinct <protein>inhibitory receptor</protein> leads to the recruitment of <protein>SHP1</protein> via its <protein>SH2 domains</protein> , activation of <protein>SHP1</protein> and subsequently inhibition of <protein>TCR</protein> signals if the <protein>inhibitory receptor</protein> is juxtaposed to the <protein>TCR</protein> .


Immune responses to adenoviruses : viral evasion mechanisms and their implications for the clinic .


Adenoviruses encode proteins that block responses to interferons , intrinsic cellular apoptosis , killing by CD8 ( + ) cytotoxic T lymphocytes and killing by the death ligands TNF , Fas ligand and TRAIL .

Adenoviruses encode proteins that block responses to <protein>interferons</protein> , intrinsic cellular apoptosis , killing by <cell_type>CD8 ( + ) cytotoxic T lymphocytes</cell_type> and killing by the <protein>death ligands</protein> <protein>TNF</protein> , <protein>Fas ligand</protein> and <protein>TRAIL</protein> .


The viral proteins are believed to prolong acute and persistent adenovirus infections .

The <protein>viral proteins</protein> are believed to prolong acute and persistent adenovirus infections .


The proteins may prove useful in protecting adenovirus gene therapy vectors and transplanted cells from the immune system .

The proteins may prove useful in protecting <dna>adenovirus gene therapy vectors</dna> and <cell_line>transplanted cells</cell_line> from the immune system .


C/EBPbeta and GATA-1 synergistically regulate activity of the eosinophil granule major basic protein promoter : implication for C/EBPbeta activity in eosinophil gene expression .

<protein>C/EBPbeta</protein> and <protein>GATA-1</protein> synergistically regulate activity of the <dna>eosinophil granule major basic protein promoter</dna> : implication for <protein>C/EBPbeta</protein> activity in eosinophil gene expression .


Eosinophil granule major basic protein ( MBP ) is expressed exclusively in eosinophils and basophils in hematopoietic cells .

<protein>Eosinophil granule major basic protein</protein> ( <protein>MBP</protein> ) is expressed exclusively in <cell_type>eosinophils</cell_type> and <cell_type>basophils</cell_type> in <cell_type>hematopoietic cells</cell_type> .


In our previous study , we demonstrated a major positive regulatory role for GATA-1 and a negative regulatory role for GATA-2 in MBP gene transcription .

In our previous study , we demonstrated a major positive regulatory role for <protein>GATA-1</protein> and a negative regulatory role for <protein>GATA-2</protein> in <protein>MBP</protein> gene transcription .


Further analysis of the MBP promoter region identified a C/EBP ( CCAAT/enhancer-binding protein ) consensus binding site 6 bp upstream of the functional GATA-binding site in the MBP gene .

Further analysis of the <dna>MBP promoter region</dna> identified a <dna>C/EBP ( CCAAT/enhancer-binding protein ) consensus binding site</dna> 6 bp upstream of the functional <dna>GATA-binding site</dna> in the <dna>MBP gene</dna> .


In the cell line HT93A , which is capable of differentiating towards both the eosinophil and neutrophil lineages in response to retinoic acid ( RA ) , C/EBPalpha mRNA expression decreased significantly concomitant with eosinophilic and neutrophilic differentiation , whereas C/EBPbeta expression was markedly increased .

In the <cell_line>cell line HT93A</cell_line> , which is capable of differentiating towards both the <cell_type>eosinophil</cell_type> and neutrophil lineages in response to retinoic acid ( RA ) , <rna>C/EBPalpha mRNA</rna> expression decreased significantly concomitant with eosinophilic and neutrophilic differentiation , whereas <protein>C/EBPbeta</protein> expression was markedly increased .


Electrophoretic mobility shift assays ( EMSAs ) showed that recombinant C/EBPbeta protein could bind to the potential C/EBP-binding site ( bp -90 to -82 ) in the MBP promoter .

Electrophoretic mobility shift assays ( EMSAs ) showed that recombinant <protein>C/EBPbeta</protein> protein could bind to the potential <dna>C/EBP-binding site</dna> ( <dna>bp -90 to -82</dna> ) in the <protein>MBP</protein> promoter .


Furthermore , we have demonstrated that both C/EBPbeta and GATA-1 can bind simultaneously to the C/EBP- and GATA-binding sites in the MBP promoter .

Furthermore , we have demonstrated that both <protein>C/EBPbeta</protein> and <protein>GATA-1</protein> can bind simultaneously to the C/EBP- and <dna>GATA-binding sites</dna> in the <dna>MBP promoter</dna> .


To determine the functionality of both the C/EBP- and GATA- binding sites , we analyzed whether C/EBPbeta and GATA-1 can stimulate the MBP promoter in the C/EBPbeta and GATA-1 negative Jurkat T-cell line .

To determine the functionality of both the <dna>C/EBP- and GATA- binding sites</dna> , we analyzed whether <protein>C/EBPbeta</protein> and <protein>GATA-1</protein> can stimulate the <dna>MBP promoter</dna> in the <cell_line>C/EBPbeta and GATA-1 negative Jurkat T-cell line</cell_line> .


Cotransfection with C/EBPbeta and GATA-1 expression vectors produced a 5-fold increase compared with cotransfection with the C/EBPbeta or GATA-1 expression vectors individually .

Cotransfection with <dna>C/EBPbeta and GATA-1 expression vectors</dna> produced a 5-fold increase compared with cotransfection with the <protein>C/EBPbeta</protein> or <dna>GATA-1 expression vectors</dna> individually .


In addition , GST pull-down experiments demonstrated a physical interaction between human GATA-1 and C/EBPbeta .

In addition , <protein>GST</protein> pull-down experiments demonstrated a physical interaction between human <protein>GATA-1</protein> and <protein>C/EBPbeta</protein> .


Expression of FOG ( riend ATA ) , which binds to GATA-1 and acts as a cofactor for GATA-binding proteins , decreased transactivation activity of GATA-1 for the MBP promoter in a dose-dependent manner .

Expression of <protein>FOG</protein> ( <protein>riend ATA</protein> ) , which binds to <protein>GATA-1</protein> and acts as a cofactor for <protein>GATA-binding proteins</protein> , decreased transactivation activity of <protein>GATA-1</protein> for the <dna>MBP promoter</dna> in a dose-dependent manner .


Our results provide the first evidence that both GATA-1 and C/EBPbeta synergistically transactivate the promoter of an eosinophil-specific granule protein gene and that FOG may act as a negative cofactor for the eosinophil lineage , unlike its positively regulatory function for the erythroid and megakaryocyte lineages .

Our results provide the first evidence that both <protein>GATA-1</protein> and <protein>C/EBPbeta</protein> synergistically transactivate the <dna>promoter</dna> of an <dna>eosinophil-specific granule protein gene</dna> and that <protein>FOG</protein> may act as a negative cofactor for the <cell_type>eosinophil lineage</cell_type> , unlike its positively regulatory function for the <cell_type>erythroid and megakaryocyte lineages</cell_type> .


Tissue-specific regulation of the ecto-5'-nucleotidase promoter .

Tissue-specific regulation of the <dna>ecto-5'-nucleotidase promoter</dna> .


Role of the camp response element site in mediating repression by the upstream regulatory region .

Role of the <dna>camp response element site</dna> in mediating repression by the <dna>upstream regulatory region</dna> .


We have isolated the 5 ' region of the ecto-5'-nucleotidase ( low K ( m ) 5'-NT ) gene and established that a 969-base pair ( bp ) fragment confers cell-specific expression of a CAT reporter gene that correlates with the expression of endogenous ecto-5'-NT mRNA and enzymatic activity .

We have isolated the <dna>5 ' region</dna> of the <dna>ecto-5'-nucleotidase ( low K ( m ) 5'-NT ) gene</dna> and established that a <dna>969-base pair ( bp ) fragment</dna> confers cell-specific expression of a <dna>CAT reporter gene</dna> that correlates with the expression of <rna>endogenous ecto-5'-NT mRNA</rna> and enzymatic activity .


A 768-bp upstream negative regulatory region has been identified that conferred lymphocyte-specific negative regulation in a heterologous system with a 244-bp deoxycytidine kinase core promoter .

A <dna>768-bp upstream negative regulatory region</dna> has been identified that conferred lymphocyte-specific negative regulation in a heterologous system with a <dna>244-bp deoxycytidine kinase core promoter</dna> .


DNase I footprinting identified several protected areas including Sp1 , Sp1/AP-2 , and cAMP response element ( CRE ) binding sites within the 201-bp core promoter region and Sp1 , NRE-2a , TCF-1/LEF-1 , and Sp1/NF-AT binding sites in the upstream regulatory region .

<protein>DNase I</protein> footprinting identified several protected areas including <dna>Sp1 , Sp1/AP-2 , and cAMP response element ( CRE ) binding sites</dna> within the 201-bp core promoter region and <dna>Sp1 , NRE-2a , TCF-1/LEF-1 , and Sp1/NF-AT binding sites</dna> in the <dna>upstream regulatory region</dna> .


Whereas the CRE site was essential in mediating the negative activity of the upstream regulatory region in Jurkat but not in HeLa cells , mutation of the Sp1/AP-2 site decreased promoter activity in both cell lines .

Whereas the <dna>CRE site</dna> was essential in mediating the negative activity of the <dna>upstream regulatory region</dna> in <cell_line>Jurkat</cell_line> but not in <cell_line>HeLa cells</cell_line> , mutation of the <dna>Sp1/AP-2 site</dna> decreased promoter activity in both cell lines .


Electrophoretic mobility shift assay analysis of proteins binding to the CRE site identified both ATF-1 and ATF-2 in Jurkat cells .

Electrophoretic mobility shift assay analysis of proteins binding to the <dna>CRE site</dna> identified both <protein>ATF-1</protein> and <protein>ATF-2</protein> in <cell_line>Jurkat cells</cell_line> .


Finally , phorbol 12-myristate 13-acetate increased the activity of both the core and the 969-bp promoter fragments , and this increase was abrogated by mutations at the CRE site .

Finally , phorbol 12-myristate 13-acetate increased the activity of both the core and the <dna>969-bp promoter fragments</dna> , and this increase was abrogated by mutations at the <dna>CRE site</dna> .


In summary , we have identified a tissue-specific regulatory region 5 ' of the ecto-5'-NT core promoter that requires the presence of a functional CRE site within the basal promoter for its suppressive activity .

In summary , we have identified a <dna>tissue-specific regulatory region</dna> 5 ' of the <dna>ecto-5'-NT core promoter</dna> that requires the presence of a functional <dna>CRE site</dna> within the <dna>basal promoter</dna> for its suppressive activity .


Extracellular signal-regulated protein kinase ( ERK ) -dependent and ERK-independent pathways target STAT3 on serine-727 in human neutrophils stimulated by chemotactic factors and cytokines .

Extracellular signal-regulated protein kinase ( ERK ) -dependent and ERK-independent pathways target <protein>STAT3</protein> on serine-727 in human neutrophils stimulated by <protein>chemotactic factors</protein> and <protein>cytokines</protein> .


STAT3 ( signal transducer and activator of transcription 3 ) is a latent transcription factor that is activated by tyrosine phosphorylation ( Tyr-705 ) in cells stimulated with cytokines or growth factors .

<protein>STAT3</protein> ( <protein>signal transducer and activator of transcription 3</protein> ) is a latent transcription factor that is activated by tyrosine phosphorylation ( Tyr-705 ) in cells stimulated with <protein>cytokines</protein> or <protein>growth factors</protein> .


Recent studies suggest that one or more cytoplasmic serine kinases also phosphorylate STAT3 and are necessary for maximal gene activation .

Recent studies suggest that one or more <protein>cytoplasmic serine kinases</protein> also phosphorylate <protein>STAT3</protein> and are necessary for maximal gene activation .


Here we demonstrate , with a site-specific antibody , that STAT3 is phosphorylated on Ser-727 in human neutrophils stimulated with chemotactic factors ( N-formyl-methionyl-leucyl-phenylalanine and complement C5a ) , cytokines [ granulocyte/macrophage colony-stimulating factor ( GM-CSF ) and granulocyte colony-stimulating factor ( G-CSF ) ] , or a protein kinase C activator ( PMA ) .

Here we demonstrate , with a site-specific antibody , that <protein>STAT3</protein> is phosphorylated on Ser-727 in <cell_type>human neutrophils</cell_type> stimulated with <protein>chemotactic factors</protein> ( N-formyl-methionyl-leucyl-phenylalanine and complement C5a ) , <protein>cytokines</protein> [ <protein>granulocyte/macrophage colony-stimulating factor</protein> ( <protein>GM-CSF</protein> ) and <protein>granulocyte colony-stimulating factor</protein> ( <protein>G-CSF</protein> ) ] , or a <protein>protein kinase C</protein> activator ( PMA ) .


( 2-Amino-3'-methoxyphenyl ) oxanaphthalen-4-one ( PD 98059 ) , an inhibitor of extracellular signal-regulated protein kinase ( ERK ) activation , blocked the serine phosphorylation of STAT3 induced by chemotactic factors or PMA .

( 2-Amino-3'-methoxyphenyl ) oxanaphthalen-4-one ( PD 98059 ) , an inhibitor of extracellular <protein>signal-regulated protein kinase</protein> ( <protein>ERK</protein> ) activation , blocked the serine phosphorylation of <protein>STAT3</protein> induced by <protein>chemotactic factors</protein> or PMA .


The drug was less effective on cytokines : it virtually abolished the response to GM-CSF that occurred 5 min after stimulation but only partly decreased those at 15-30 min and did not appreciably alter responses to G-CSF regardless of incubation time .

The drug was less effective on <protein>cytokines</protein> : it virtually abolished the response to <protein>GM-CSF</protein> that occurred 5 min after stimulation but only partly decreased those at 15-30 min and did not appreciably alter responses to <protein>G-CSF</protein> regardless of incubation time .


1- ( 5-Isoquinolinylsulphonyl ) -2-methylpiperazine dihydrochloride ( H7 ) , an inhibitor of a putative STAT3 serine kinase , and 4- ( 4-fluorophenyl ) -2- ( 4-methylsulphinylphenyl ) -5- ( 4-pyridyl ) 1H-imidazole ( SB 203580 ) , an inhibitor of p38 mitogen-activated protein ( MAP ) kinase , did not dampen any of these serine phosphorylation responses .

1- ( 5-Isoquinolinylsulphonyl ) -2-methylpiperazine dihydrochloride ( H7 ) , an inhibitor of a putative <protein>STAT3 serine kinase</protein> , and 4- ( 4-fluorophenyl ) -2- ( 4-methylsulphinylphenyl ) -5- ( 4-pyridyl ) 1H-imidazole ( SB 203580 ) , an inhibitor of <protein>p38 mitogen-activated protein ( MAP ) kinase</protein> , did not dampen any of these serine phosphorylation responses .


We propose that neutrophils use both ERK-dependent and ERK-independent pathways to phosphorylate Ser-727 on STAT3 .

We propose that <cell_type>neutrophils</cell_type> use both ERK-dependent and ERK-independent pathways to phosphorylate Ser-727 on <protein>STAT3</protein> .


The former pathway is recruited by all ERK -activating stimuli , whereas the latter pathway uses an undefined serine kinase and is recruited selectively by cytokines .

The former pathway is recruited by all <protein>ERK</protein> -activating stimuli , whereas the latter pathway uses an undefined <protein>serine kinase</protein> and is recruited selectively by <protein>cytokines</protein> .


GATA transcription factors associate with a novel class of nuclear bodies in erythroblasts and megakaryocytes .

<protein>GATA transcription factors</protein> associate with a novel class of nuclear bodies in <cell_type>erythroblasts</cell_type> and <cell_type>megakaryocytes</cell_type> .


The nuclear distribution of GATA transcription factors in murine haemopoietic cells was examined by indirect immunofluorescence .

The nuclear distribution of <protein>GATA transcription factors</protein> in <cell_type>murine haemopoietic cells</cell_type> was examined by indirect immunofluorescence .


Specific bright foci of GATA-1 fluorescence were observed in erythroleukaemia cells and primary murine erythroblasts and megakaryocytes , in addition to diffuse nucleoplasmic localization .

Specific bright foci of <protein>GATA-1</protein> fluorescence were observed in <cell_type>erythroleukaemia cells</cell_type> and <cell_type>primary murine erythroblasts</cell_type> and <cell_type>megakaryocytes</cell_type> , in addition to diffuse nucleoplasmic localization .


These foci , which were preferentially found adjacent to nucleoli or at the nuclear periphery , did not represent sites of active transcription or binding of GATA-1 to consensus sites in the beta-globin loci .

These foci , which were preferentially found adjacent to nucleoli or at the nuclear periphery , did not represent sites of active transcription or binding of <protein>GATA-1</protein> to <dna>consensus sites</dna> in the <dna>beta-globin loci</dna> .


Immunoelectron microscopy demonstrated the presence of intensely labelled structures likely to represent the GATA-1 foci seen by immunofluorescence .

Immunoelectron microscopy demonstrated the presence of intensely labelled structures likely to represent the <protein>GATA-1</protein> foci seen by immunofluorescence .


The GATA-1 nuclear bodies differed from previously described nuclear structures and there was no co-localization with nuclear antigens involved in RNA processing or other ubiquitous ( Spl , c-Jun and TBP ) or haemopoietic ( NF-E2 ) transcription factors .

The <protein>GATA-1</protein> nuclear bodies differed from previously described nuclear structures and there was no co-localization with <protein>nuclear antigens</protein> involved in RNA processing or other <protein>ubiquitous ( Spl , c-Jun and TBP ) or haemopoietic ( NF-E2 ) transcription factors</protein> .


Interestingly , GATA-2 and GATA-3 proteins also localized to the same nuclear bodies in cell lines co-expressing GATA-1 and -2 or GATA-1 and -3 gene products .

Interestingly , <protein>GATA-2</protein> and <protein>GATA-3 proteins</protein> also localized to the same nuclear bodies in cell lines co-expressing <protein>GATA-1 and -2 or GATA-1 and -3 gene products</protein> .


This pattern of distribution is , thus far , unique to the GATA transcription factors and suggests a protein-protein interaction with other components of the nuclear bodies via the GATA zinc finger domain .

This pattern of distribution is , thus far , unique to the <protein>GATA transcription factors</protein> and suggests a protein-protein interaction with other components of the nuclear bodies via the <protein>GATA zinc finger domain</protein> .


Energy substrates , hormone responses and glucocorticoid binding in lymphocytes during intense physical exercise in humans following phosphocreatine administration .

Energy substrates , hormone responses and glucocorticoid binding in <cell_type>lymphocytes</cell_type> during intense physical exercise in humans following phosphocreatine administration .


Eight healthy untrained male volunteers pedalled a cycle ergometer according to two exercise protocols : the first involved step-wise increasing physical exercise to maximal ( MPE ) ; the second involved prolonged ( 35 min ) submaximal physical exercise ( PPE ) at 70 % of the individual 's maximal oxygen uptake .


Each volunteer performed these exercise twice , following either an intravenous injection of phosphocreatine ( PCr ) or a placebo of an isotonic NaCl solution .


Anaerobic threshold ( AT ) was determined from the point of departure of the ventilatory response from linearity and from the sudden increase in venous blood lactate concentrations during MPE .


After exercise following placebo administration we observed increases in concentrations of blood substrates , plasma adrenocorticotropin ( ACTH ) , growth hormone and cortisol and in the number of glucocorticoid receptors in lymphocytes without changes in the dissociation constant .

After exercise following placebo administration we observed increases in concentrations of blood substrates , <protein>plasma adrenocorticotropin</protein> ( <protein>ACTH</protein> ) , growth hormone and cortisol and in the number of <protein>glucocorticoid receptors</protein> in <cell_type>lymphocytes</cell_type> without changes in the dissociation constant .


Intravenous administration of PCr ( starting 1 day before exercise ) led to an increase in the total workload ( on average by 5.8 % ) and in AT ( on average by 6.8 % ) during MPE and to a better tolerance of exercise during PPE .


Following PCr administration we observed lower blood lactate concentrations and different patterns of some enzyme activities , less pronounced changes in plasma ACTH and cortisol concentrations and in glucocorticoid binding in lymphocytes , but no changes in plasma growth hormone concentrations compared to the placebo .

Following PCr administration we observed lower blood lactate concentrations and different patterns of some enzyme activities , less pronounced changes in <protein>plasma ACTH</protein> and cortisol concentrations and in glucocorticoid binding in <cell_type>lymphocytes</cell_type> , but no changes in plasma growth hormone concentrations compared to the placebo .


The results showed that intense physical exercise led not only to increases in blood hormone concentrations but also to an increase in the density of glucocorticoid receptors in lymphocytes .

The results showed that intense physical exercise led not only to increases in blood hormone concentrations but also to an increase in the density of <protein>glucocorticoid receptors</protein> in <cell_type>lymphocytes</cell_type> .


Intravenous PCr injection led to smaller changes in ACTH and cortisol concentrations as well as to a lower activation of glucocorticoid binding in lymphocytes .

Intravenous PCr injection led to smaller changes in <protein>ACTH</protein> and cortisol concentrations as well as to a lower activation of glucocorticoid binding in <cell_type>lymphocytes</cell_type> .


Putative steroid binding domain of the human mineralocorticoid receptor , expressed in E. coli in the presence of heat shock proteins shows typical native receptor characteristics .

<protein>Putative steroid binding domain</protein> of the <protein>human mineralocorticoid receptor</protein> , expressed in E. coli in the presence of <protein>heat shock proteins</protein> shows typical native receptor characteristics .


Domain E , considered as the putative hormone binding domain ( HBD ) of the human mineralocorticoid receptor ( hMR ) was expressed in Escherichia coli as a fusion protein with either maltose binding protein ( MBP ) or glutathione S-transferase ( GST ) .

<protein>Domain E</protein> , considered as the <protein>putative hormone binding domain</protein> ( <protein>HBD</protein> ) of the <protein>human mineralocorticoid receptor</protein> ( <protein>hMR</protein> ) was expressed in Escherichia coli as a fusion protein with either <protein>maltose binding protein</protein> ( <protein>MBP</protein> ) or <protein>glutathione S-transferase</protein> ( <protein>GST</protein> ) .


These bacterially-produced MR constructs had no steroid binding activity per se .

These <dna>bacterially-produced MR constructs</dna> had no steroid binding activity per se .


In fact , heat shock protein association ( hsp ) is required for high affinity ligand-binding of the MR .

In fact , <protein>heat shock protein</protein> association ( <protein>hsp</protein> ) is required for high affinity ligand-binding of the <protein>MR</protein> .


After incubation of purified MBP- or GST-HBD with rabbit reticulocyte lysate , known to be rich in heat shock proteins , we obtained saturable binding of [ 3H ] aldosterone .

After incubation of purified <protein>MBP- or GST-HBD</protein> with rabbit reticulocyte lysate , known to be rich in <protein>heat shock proteins</protein> , we obtained saturable binding of [ 3H ] aldosterone .


The Kd value for aldosterone was 0.3 nM and the Bmax = 32 pmol/mg .


Hormone binding specificity was assessed by competition studies with various steroid ligands .


Sucrose gradient assays performed with [ 3H ] aldosterone-MBP-HBD revealed complex sedimenting at 8.3S and 4.9S with [ 3H ] progesterone-MBP-HBD .

Sucrose gradient assays performed with <protein>[ 3H ] aldosterone-MBP-HBD</protein> revealed complex sedimenting at 8.3S and 4.9S with <protein>[ 3H ] progesterone-MBP-HBD</protein> .


Western-blot analysis of the sedimentation peak showed the concomitant presence of MBP-HBD by a monoclonal anti-MBP antibody , and hsp90 by a monoclonal anti-hsp antibody .

Western-blot analysis of the sedimentation peak showed the concomitant presence of <protein>MBP-HBD</protein> by a <protein>monoclonal anti-MBP antibody</protein> , and <protein>hsp90</protein> by a <protein>monoclonal anti-hsp antibody</protein> .


Moreover , following incubation with the anti-rabbit hsp90 monoclonal antibody the sedimenting gradient showed a 10.4S sedimenting complex .

Moreover , following incubation with the <protein>anti-rabbit hsp90 monoclonal antibody</protein> the sedimenting gradient showed a <protein>10.4S sedimenting complex</protein> .


These analyses demonstrated that the [ 3H ] aldosterone- MBP-HBD complex is at least associated with hsp90 in reticulocyte lysate and that the HBD of hMR is sufficient to bind hsp90 .

These analyses demonstrated that the [ 3H ] aldosterone- <protein>MBP-HBD complex</protein> is at least associated with <protein>hsp90</protein> in reticulocyte lysate and that the <protein>HBD</protein> of <protein>hMR</protein> is sufficient to bind <protein>hsp90</protein> .


Deletions of a relatively short amino- ( 729-766 ) or carboxy- terminal ( 940-984 ) region of the HBD fragment eliminated all steroid-binding properties .

Deletions of a relatively short <protein>amino- ( 729-766 ) or carboxy- terminal ( 940-984 ) region</protein> of the <protein>HBD fragment</protein> eliminated all steroid-binding properties .


Overall , these results indicate that the integrity of domain E is necessary and sufficient to bind steroid ligands , agonists or antagonists , with characteristics similar to the entire native MR .

Overall , these results indicate that the integrity of <protein>domain E</protein> is necessary and sufficient to bind steroid ligands , agonists or antagonists , with characteristics similar to the entire native <protein>MR</protein> .


Renal cell carcinoma-derived gangliosides suppress nuclear factor-kappaB activation in T cells .

Renal cell carcinoma-derived gangliosides suppress <protein>nuclear factor-kappaB</protein> activation in <cell_type>T cells</cell_type> .


Activation of the transcription factor nuclear factor-kappaB ( NFkappaB ) is impaired in T cells from patients with renal cell carcinomas ( RCCs ) .

Activation of the <protein>transcription factor nuclear factor-kappaB</protein> ( <protein>NFkappaB</protein> ) is impaired in <cell_type>T cells</cell_type> from patients with renal cell carcinomas ( RCCs ) .


In circulating T cells from a subset of patients with RCCs , the suppression of NFkappaB binding activity is downstream from the stimulus-induced degradation of the cytoplasmic factor IkappaBalpha .

In circulating <cell_type>T cells</cell_type> from a subset of patients with RCCs , the suppression of <protein>NFkappaB</protein> binding activity is downstream from the stimulus-induced degradation of the <protein>cytoplasmic factor</protein> <protein>IkappaBalpha</protein> .


Tumor-derived soluble products from cultured RCC explants inhibit NFkappaB activity in T cells from healthy volunteers , despite a normal level of stimulus-induced IkappaBalpha degradation in these cells .

<protein>Tumor-derived soluble products</protein> from cultured RCC explants inhibit <protein>NFkappaB</protein> activity in <cell_type>T cells</cell_type> from healthy volunteers , despite a normal level of stimulus-induced <protein>IkappaBalpha</protein> degradation in these cells .


The inhibitory agent has several features characteristic of a ganglioside , including sensitivity to neuraminidase but not protease treatment ; hydrophobicity ; and molecular weight less than 3 kDa .

The inhibitory agent has several features characteristic of a ganglioside , including sensitivity to <protein>neuraminidase</protein> but not <protein>protease</protein> treatment ; hydrophobicity ; and molecular weight less than 3 kDa .


Indeed , we detected gangliosides in supernatants from RCC explants and not from adjacent normal kidney tissue .

Indeed , we detected gangliosides in supernatants from <cell_type>RCC explants</cell_type> and not from adjacent normal kidney tissue .


Gangliosides prepared from RCC supernatants , as well as the purified bovine gangliosides G ( m1 ) and G ( d1a ) , suppressed NFkappaB binding activity in T cells and reduced expression of the cytokines IL-2 and IFN-gamma .

Gangliosides prepared from RCC supernatants , as well as the purified bovine gangliosides G ( m1 ) and G ( d1a ) , suppressed <protein>NFkappaB</protein> binding activity in <cell_type>T cells</cell_type> and reduced expression of the <protein>cytokines</protein> <protein>IL-2</protein> and <protein>IFN-gamma</protein> .


Taken together , our findings suggest that tumor-derived gangliosides may blunt antitumor immune responses in patients with RCCs .


Nuclear factor-kappaB -dependent induction of interleukin-8 gene expression by tumor necrosis factor alpha : evidence for an antioxidant sensitive activating pathway distinct from nuclear translocation .

<protein>Nuclear factor-kappaB</protein> -dependent induction of <protein>interleukin-8</protein> gene expression by <protein>tumor necrosis factor alpha</protein> : evidence for an antioxidant sensitive activating pathway distinct from nuclear translocation .


Tumor necrosis factor alpha ( TNFalpha ) is a pluripotent activator of inflammation by inducing a proinflammatory cytokine cascade .

<protein>Tumor necrosis factor alpha</protein> ( <protein>TNFalpha</protein> ) is a pluripotent activator of inflammation by inducing a <protein>proinflammatory cytokine</protein> cascade .


This phenomenon is mediated , in part , through inducible expression of the CXC chemokine , interleukin-8 ( IL-8 ) .

This phenomenon is mediated , in part , through inducible expression of the <protein>CXC chemokine</protein> , <protein>interleukin-8</protein> ( IL-8 ) .


In this study , we investigate the role of TNFalpha -inducible reactive oxygen species ( ROS ) in IL-8 expression by `` monocyte-like '' U937 histiocytic lymphoma cells .

In this study , we investigate the role of <protein>TNFalpha</protein> -inducible reactive oxygen species ( ROS ) in <protein>IL-8</protein> expression by `` monocyte-like '' <cell_line>U937 histiocytic lymphoma cells</cell_line> .


TNFalpha is a rapid activator of IL-8 gene expression by U937 , producing a 50-fold induction of mRNA within 1 hour of treatment .

<protein>TNFalpha</protein> is a rapid activator of <dna>IL-8 gene</dna> expression by <cell_line>U937</cell_line> , producing a 50-fold induction of <rna>mRNA</rna> within 1 hour of treatment .


In gene transfection assays , the effect of TNFalpha requires the presence of an inducible nuclear factor-kappaB ( NF-kappaB ) ( Rel A ) binding site in the IL-8 promoter .

In gene transfection assays , the effect of <protein>TNFalpha</protein> requires the presence of an <dna>inducible nuclear factor-kappaB ( NF-kappaB ) ( Rel A ) binding site</dna> in the <dna>IL-8 promoter</dna> .


TNFalpha treatment induces a rapid translocation of the 65 kD transcriptional activator NF-kappaB subunit , Rel A , whose binding in the nucleus occurs before changes in intracellular ROS .

<protein>TNFalpha</protein> treatment induces a rapid translocation of the <protein>65 kD transcriptional activator</protein> <protein>NF-kappaB subunit</protein> , <protein>Rel A</protein> , whose binding in the nucleus occurs before changes in intracellular ROS .


Pretreatment ( or up to 15 minutes posttreatment ) relative to TNFalpha with the antioxidant dimethyl sulfoxide ( DMSO ) ( 2 % [ vol/vol ] ) blocks 80 % of NF-kappaB -dependent transcription .

Pretreatment ( or up to 15 minutes posttreatment ) relative to <protein>TNFalpha</protein> with the antioxidant dimethyl sulfoxide ( DMSO ) ( 2 % [ vol/vol ] ) blocks 80 % of <protein>NF-kappaB</protein> -dependent transcription .


Surprisingly , however , DMSO has no effect on inducible Rel A binding .

Surprisingly , however , DMSO has no effect on inducible <protein>Rel A</protein> binding .


Similar selective effects on NF-kappaB transcription are seen with the unrelated antioxidants , N-acetylcysteine ( NAC ) and vitamin C .

Similar selective effects on <protein>NF-kappaB</protein> transcription are seen with the unrelated antioxidants , N-acetylcysteine ( NAC ) and vitamin C .


These data indicate that TNFalpha induces a delayed ROS-dependent signalling pathway that is required for NF-kappaB transcriptional activation and is separable from that required for its nuclear translocation .

These data indicate that <protein>TNFalpha</protein> induces a delayed ROS-dependent signalling pathway that is required for <protein>NF-kappaB</protein> transcriptional activation and is separable from that required for its nuclear translocation .


Further definition of this pathway will yield new insights into inflammation initiated by TNFalpha signalling .

Further definition of this pathway will yield new insights into inflammation initiated by <protein>TNFalpha</protein> signalling .


Gender and vascular reactivity .


Estrogen receptors are found on vascular endothelial and smooth muscle cells ; their expression is influenced by exposure to the hormone .

<protein>Estrogen receptors</protein> are found on <cell_type>vascular endothelial and smooth muscle cells</cell_type> ; their expression is influenced by exposure to the hormone .


Estrogen receptors influence non-genomic events , which are rapid in onset and genomic events , which are longer acting responses .

<protein>Estrogen receptors</protein> influence non-genomic events , which are rapid in onset and genomic events , which are longer acting responses .


Estrogens affect vascular tone indirectly by modulating release of endothelium-derived vasoactive factors and directly by modulating intracellular calcium in vascular smooth muscle cells .

Estrogens affect vascular tone indirectly by modulating release of <protein>endothelium-derived vasoactive factors</protein> and directly by modulating intracellular calcium in <cell_type>vascular smooth muscle cells</cell_type> .


Estrogens indirectly affect thrombotic events and inflammation by altering platelet aggregation and leukocyte adherence and migration , respectively .

Estrogens indirectly affect thrombotic events and inflammation by altering <cell_type>platelet</cell_type> aggregation and <cell_type>leukocyte</cell_type> adherence and migration , respectively .


Estrogens also influence production of mitogens which , when released at sites of vascular injury , affect vascular remodeling .


Although estrogens initiate vascular responses , genomic sex may influence and/or limit expression of estrogen receptors and therefore actions of sex steroid hormones throughout the vasculature .


Induction of a functional vitamin D receptor in all-trans-retinoic acid-induced monocytic differentiation of M2-type leukemic blast cells .

Induction of a functional <protein>vitamin D receptor</protein> in all-trans-retinoic acid-induced monocytic differentiation of <cell_line>M2-type leukemic blast cells</cell_line> .


Different types of acute myeloid leukemia blast cells were induced to differentiate in vitro with all-trans-retinoic acid ( ATRA ) and vitamin D3 ( VD ) .

Different types of <cell_type>acute myeloid leukemia blast cells</cell_type> were induced to differentiate in vitro with all-trans-retinoic acid ( ATRA ) and vitamin D3 ( VD ) .


M0/M1 leukemic cells are not sensitive to differentiating agents , whereas M3 leukemic cells are induced to undergo granulocytic differentiation after ATRA treatment but are not sensitive to VD .

<cell_line>M0/M1 leukemic cells</cell_line> are not sensitive to differentiating agents , whereas <cell_line>M3 leukemic cells</cell_line> are induced to undergo granulocytic differentiation after ATRA treatment but are not sensitive to VD .


M2 leukemic blast cells behave differently because they undergo monocytic differentiation with both the differentiation inducers .

<cell_line>M2 leukemic blast cells</cell_line> behave differently because they undergo monocytic differentiation with both the differentiation inducers .


To gain some insight into the maturation of M2-type leukemic cells , we studied the molecular mechanisms underlying monocytic differentiation induced by ATRA and VD in spontaneous M2 blast cells as well as in Kasumi-1 cells ( an acute myeloid leukemia M2-type cell line ) .

To gain some insight into the maturation of <cell_type>M2-type leukemic cells</cell_type> , we studied the molecular mechanisms underlying monocytic differentiation induced by ATRA and VD in spontaneous <cell_line>M2 blast cells</cell_line> as well as in <cell_line>Kasumi-1 cells</cell_line> ( an <cell_line>acute myeloid leukemia M2-type cell line</cell_line> ) .


Our results indicate that ATRA as well as VD efficiently increases the nuclear abundance of VD receptor ( VDR ) and promotes monocytic differentiation .

Our results indicate that ATRA as well as VD efficiently increases the nuclear abundance of <protein>VD receptor</protein> ( <protein>VDR</protein> ) and promotes monocytic differentiation .


VDR is functionally active in ATRA-treated Kasumi-1 cells because it efficiently heterodimerizes with retinoid X receptor , binds to a DR3-type vitamin D-responsive element , and activates the transcription of a vitamin D-responsive element-regulated reporter gene .

<protein>VDR</protein> is functionally active in ATRA-treated <cell_line>Kasumi-1 cells</cell_line> because it efficiently heterodimerizes with <protein>retinoid X receptor</protein> , binds to a <dna>DR3-type vitamin D-responsive element</dna> , and activates the transcription of a <dna>vitamin D-responsive element-regulated reporter gene</dna> .


Consistent with these findings , VD-responsive genes are induced by ATRA treatment of Kasumi-1 cells , suggesting that the genetic program underlying monocytic differentiation is activated .

Consistent with these findings , <dna>VD-responsive genes</dna> are induced by ATRA treatment of <cell_line>Kasumi-1 cells</cell_line> , suggesting that the genetic program underlying monocytic differentiation is activated .


The molecular mechanism by which ATRA increases the nuclear abundance of a functional VDR is still unknown , but our data clearly indicate that the M2 leukemic cell context is only permissive of monocytic differentiation .

The molecular mechanism by which ATRA increases the nuclear abundance of a functional <protein>VDR</protein> is still unknown , but our data clearly indicate that the <cell_line>M2 leukemic cell</cell_line> context is only permissive of monocytic differentiation .


Protein kinase C and calcineurin synergize to activate IkappaB kinase and NF-kappaB in T lymphocytes .

<protein>Protein kinase C</protein> and <protein>calcineurin</protein> synergize to activate <protein>IkappaB kinase</protein> and <protein>NF-kappaB</protein> in <cell_type>T lymphocytes</cell_type> .


The nuclear factor of kappaB ( NF-kappaB ) is a ubiquitous transcription factor that is key in the regulation of the immune response and inflammation .

The nuclear factor of kappaB ( <protein>NF-kappaB</protein> ) is a <protein>ubiquitous transcription factor</protein> that is key in the regulation of the immune response and inflammation .


T cell receptor ( TCR ) cross-linking is in part required for activation of NF-kappaB , which is dependent on the phosphorylation and degradation of IkappaBalpha .

<protein>T cell receptor</protein> ( <protein>TCR</protein> ) cross-linking is in part required for activation of <protein>NF-kappaB</protein> , which is dependent on the phosphorylation and degradation of <protein>IkappaBalpha</protein> .


By using Jurkat and primary human T lymphocytes , we demonstrate that the simultaneous activation of two second messengers of the TCR -initiated signal transduction , protein kinase C ( PKC ) and calcineurin , results in the synergistic activation of the IkappaBalpha kinase ( IKK ) complex but not of another putative IkappaBalpha kinase , p90 ( rsk ) .

By using <cell_line>Jurkat</cell_line> and <cell_type>primary human T lymphocytes</cell_type> , we demonstrate that the simultaneous activation of two second messengers of the <protein>TCR</protein> -initiated signal transduction , <protein>protein kinase C</protein> ( <protein>PKC</protein> ) and <protein>calcineurin</protein> , results in the synergistic activation of the <protein>IkappaBalpha kinase ( IKK ) complex</protein> but not of another <protein>putative IkappaBalpha kinase</protein> , <protein>p90 ( rsk )</protein> .


We also demonstrate that the IKK complex , but not p90 ( rsk ) , is responsible for the in vivo phosphorylation of IkappaBalpha mediated by the co-activation of PKC and calcineurin .

We also demonstrate that the <protein>IKK complex</protein> , but not <protein>p90 ( rsk )</protein> , is responsible for the in vivo phosphorylation of <protein>IkappaBalpha</protein> mediated by the co-activation of <protein>PKC</protein> and <protein>calcineurin</protein> .


Each second messenger is necessary , as inhibition of either one reverses the activation of the IKK complex and IkappaBalpha phosphorylation in vivo .

Each second messenger is necessary , as inhibition of either one reverses the activation of the <protein>IKK complex</protein> and <protein>IkappaBalpha</protein> phosphorylation in vivo .