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Biomedical subjects

R Girard

Publications and source records attributed to R Girard.

At least 37 records · Page 2Linked to original sources
PubMed1999

Exogenous cyclic AMP, cholera toxin, and endotoxin induce expression of the lipopolysaccharide receptor CD14 in murine bone marrow cells: role of purinoreceptors.

Little is known about the mechanisms of lipopolysaccharide (LPS) signaling in immature cells that do not express the LPS receptor CD14 yet. Bone marrow granulocytes do not constitutively express CD14 but can be stimulated by low doses of LPS in the absence of serum and then express an inducible form of LPS receptor (iLpsR). We show that in addition to LPS, cholera toxin (CT) and various cyclic AMP (cAMP) analogs can also induce the expression of iLpsR, which was identified as CD14. Induction was independent of intracellular cAMP. The hypothesis that cAMP analogs act via a cell surface receptor was suggested by the unresponsiveness of trypsin-treated cells to these inducers and by the specific binding of [(3)H]cAMP to the cells. This binding was not inhibited by LPS or CT but was inhibited by various purine derivatives. However, the receptor involved is not a conventional purinoreceptor since both an agonist and an antagonist of such receptors were able to induce iLpsR expression. The results suggest that cAMP analogs and other purine derivatives induce iLpsR after interaction with an unconventional, trypsin-sensitive, purinoreceptor distinct from LPS and CT receptors.

1-Methyl-3-isobutylxanthine↗
PubMed1999

Interaction of NK lysin, a peptide produced by cytolytic lymphocytes, with endotoxin.

NK lysin is a 9-kDa polypeptide that was originally isolated from porcine intestinal tissue based on its antibacterial activity. It is produced by cytolytic lymphocytes and is cytolytic against a number of different types of tumor cells. Here we report the binding of NK lysin to lipopolysaccharide (LPS) and its anti-LPS activity. NK lysin binds to matrix-coated LPS from Escherichia coli, Pseudomonas aeruginosa, and different strains of Salmonella enterica. Lipid A and polymyxin B inhibited the binding, demonstrating a preferential interaction of NK lysin with the lipid part of LPS. Chromium-labeled lymphoma cells were lysed by NK lysin, and LPS dose-dependently inhibited the cytolysis at equimolar amounts. In the same manner, NK lysin inhibited certain LPS-stimulated effects on mouse bone marrow cells as well as LPS binding to mouse granulocytes. These results suggest that NK lysin may be a another natural LPS-binding protein from lymphocytes that may participate in the endogenous defense response associated with elevated concentrations of LPS.

Animals↗
PubMed1997

Facial mask noninvasive mechanical ventilation reduces the incidence of nosocomial pneumonia. A prospective epidemiological survey from a single ICU.

OBJECTIVE: To evaluate the impact of noninvasive positive pressure mechanical ventilation (NPPV) on ventilator-associated pneumonia (VAP). DESIGN: Prospective observational study. SETTING: Medical intensive care unit (ICU) of a university teaching hospital. PATIENTS: Cohort of 320 consecutive patients staying in the ICU more than 2 days and mechanically ventilated for > or = 1 day. MEASUREMENTS AND RESULTS: VAP was diagnosed when, satisfying classical clinical and radiological criteria, fiberoptic bronchoalveolar lavage and/or protected specimen brush grew > or = 10(4) and > or = 10(3) CFU/ml, respectively, of at least one microorganism. Patients were classified into four subgroups according to the way in which mechanical ventilation was delivered: NPPV then tracheal intubation (TI) (n = 38), TI then NPPV (n = 23), TI only (n = 199), and NPPV only (n = 60). Occurrence of VAP was estimated by incidence rate and density of incidence. Risk factors for VAP were assessed by logistic regression analysis. Twenty-seven patients had 28 episodes of VAP. The incidence rates for patients with VAP were 18% in NPPV-TI, 22% in TI-NPPV, 8% in TI, and 0% in NPPV (p < 0.0001). The density of incidence of VAP was 0.85 per 100 days of TI and 0.16 per 100 days of NPPV (p = 0.04). Logistic regression showed that length of ICU stay and ventilatory support were associated with VAP. CONCLUSIONS: There is a significantly lower incidence of VAP associated with NPPV compared to tracheal intubation. This is mainly explained by differences in patient severity and risk exposure.

APACHE↗
PubMed1997

Functional lipopolysaccharide receptors of low affinity are constitutively expressed on mouse bone marrow cells.

Although lipopolysaccharide (LPS)-induced overproduction of cytokines, involved in the pathogenesis of septic shock, occupies the spotlight of endotoxin research, another LPS effect, the differentiation of various cell types including haematopoietic bone marrow cells (BMC), which is probably related to its radioprotective activity, deserves equal attention. We have previously established that nanomolar concentrations of LPS trigger in human BMC the expression of CD14 by an induction mechanism independent of CD14 or any other molecule anchored to the cell membrane by a glycosyl phosphatidylinositol glycolipid. We now show that this LPS-induced stimulation is triggered by the binding of a small number of LPS molecules (13,000 molecules/cell) to constitutive LPS receptors of low affinity (Kd = 480 nM). This interaction, which was inhibited by a synthetic LPS antagonist, appeared specific, reversible, saturable, time- and temperature-dependent, but was independent of divalent cations, and was inhibited by serum. Exposure of BMC to LPS did not induce a down-modulation of these receptors, but enhanced their sensitivity to trypsin degradation. Inhibition of LPS binding following different treatments correlated with inhibition of BMC stimulation, thus suggesting that the sparse constitutive receptors of low affinity are efficient signalling receptors for LPS.

Animals↗
PubMed1997

Lipopolysaccharide and the glycoside ring of staurosporine induce CD14 expression on bone marrow granulocytes by different mechanisms.

We established previously that lipopolysaccharide (LPS) can induce the expression of LPS-binding sites on bone marrow cells (BMC). We now report that staurosporine (STP), a glycosylated indolocarbazole alkaloid with potent inhibitory activity for various protein kinases, can induce the same effect. With both agents, the newly expressed LPS receptor was found to be CD14. The STP-induced effect was independent of its protein kinase inhibitory activity because several other protein kinase inhibitors, such as the indolocarbazole K-252a, the bisindolylmaleimide RO-31-8220, the perylenequinone calphostin C, and the isoquinolinesulfonamide H7, did not induce CD14 expression. The observation that the STP analog K-252a with an identical polyaromatic aglycon moiety was inactive yet the analog UCN-01 with an identical glycoside ring was active suggests that the induction of CD14 expression is triggered by the sugar moiety of STP. Three lines of evidence show that the mechanism of CD14 expression induced by STP differs from that induced by LPS: (i) unlike LPS, STP can stimulate BMC from LPS-unresponsive C3H/HeJ mice, (ii) LPS and STP effects are additive at a saturating dose of LPS, and (iii) the protein kinase inhibitor K-252a inhibits the LPS-induced but not STP-induced stimulation. Therefore, our findings show that both a protein kinase-dependent (LPS-induced) and a protein kinase-independent (STP-induced) mechanism can lead to the expression of the LPS receptor CD14 on BMC. We also found that the STP-induced stimulation of BMC is modulated by cyclosporin A, vinblastine, and verapamil. This observation may suggest that the inducible effect of STP could be initiated by its interaction with P-glycoprotein, a membrane pump with drug efflux function that plays a critical role in the multidrug resistance of cancer cells.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1996

The putative immunity protein of the gram-positive bacteria Leuconostoc mesenteroides is preferentially located in the cytoplasm compartment.

Immunity proteins are though to protect bacteriocin-producing bacterial strains against the bactericidal effects of their own bacteriocin. The immunity protein which protects the lactic acid bacterium Leuconostoc mesenteroides against mesentericin Y105(37) bacteriocin was detected and localized by immunofluorescence and electron microscopy, using antibodies directed against the C-terminal end of the predicted immunity protein. The antibodies recognized the immunity proteins of various strains of Leuconostoc, including Leuconostoc mesenteroides and Leuconostoc gelidum. This study demonstrated that immunity proteins produced by Leuconostoc mesenteroides accumulated in the cytoplasmic compartment of the bacteria. This is in contrast with other known immunity proteins, such as the colicin immunity proteins, which are integral membrane proteins possessing three to four transmembrane domains.

Amino Acid Sequence↗
PubMed1996

Inability of the Francisella tularensis lipopolysaccharide to mimic or to antagonize the induction of cell activation by endotoxins.

We studied the ability of the lipopolysaccharide (LPS) extracted from a vaccine strain of Francisella tularensis (LPS-Ft) to mimic LPSs from other gram-negative bacteria for activation of various murine cell types or to antagonize the effects of other LPSs. We found that activation of macrophages for the production of tumor necrosis factor alpha and NO, of pre-B lymphocytes for the expression of surface immunoglobulins, and of bone marrow cells for the expression of LPS-binding sites was either undetectable with LPS-Ft or required concentrations 100 to 1,000 times higher than for standard LPSs. Preexposure of macrophages to LPS-Ft also failed to trigger down-regulation of tumor necrosis factor alpha (desensitization) or up-regulation of NO responses to an endotoxin challenge. In contrast to other atypical LPSs, LPS-Ft was also unable to antagonize any of the endotoxin-induced cellular responses mentioned above, suggesting that this LPS does not interact with LPS receptors.

Animals↗
PubMed1995

Variation of LPS-binding capacity, epitope expression, and shedding of membrane-bound CD14 during differentiation of human monocytes.

The myeloid differentiation Ag CD14 is considered to play a critical role in the binding of LPS to monocytes. To determine if differences in LPS-binding capacities of cells could reflect a variability of CD14 molecules, we analyzed the interactions of various reagents with these molecules in human blood monocytes and in promyelocytic (HL60) and monocytic (THP-1) cell lines. The expression of CD14 epitopes was analyzed with the fluorescent anti-CD14 mAbs My4 and LeuM3. Expression of LPS-binding sites (LPS+ molecules) was detected with LPS-FITC. THP-1 cells stimulated with calcitriol (VitD3), as well as the majority of blood monocytes (50-90%) were My4+/LPS+. However, untreated THP-1 cells, and a substantial population (10-50%) of human monocytes from healthy donors, were My4+/LPS-, thus suggesting the existence of CD14 isoforms with different LPS-binding capacities. In line with this assumption, monocytes stimulated with PMA selectively shed LeuM3+ molecules, but almost no My4+ and LPS+ constituents. Analysis of monocytes after treatment with phosphatidylinositol-specific phospholipase C indicated that among CD14 molecules with LPS-binding capacity, some are susceptible and others are resistant to the enzyme, each type being mainly expressed by a different monocyte subset. Studies of uninduced and chemically induced THP-1 cells showed that wheat-germ agglutinin blocked the binding of My4 to constitutive, but not to chemically inducible CD14. The overall results suggest the existence of at least three different forms of CD14, which may reflect different stages of cell maturation.

Adult↗
PubMed1995

Dielectric properties of grapes and sugar solutions at 2.45 GHz.

Dielectric properties of grapes at 2.45 GHz were measured at moisture contents varying from 80 to 15% (wet basis) at temperatures ranging from 25-80 degrees C using the Open Ended Coaxial Transmission Line Technique. Both dielectric constant and loss factor decreased with decreasing moisture content. In low moisture samples these properties were found to be considerably influenced by higher temperatures. Dielectric properties of sugar solutions of varying concentrations (20-250% by weight in water) were also determined at different temperatures, and these values were compared to those of grapes of corresponding moisture concentration. Predictive models of the dieletric properties as functions of moisture content and temperature were generated using response surface methodology. The results are useful in estimating the volumetric heating of grapes by microwave energy, and these methods can be extended to sugar-based foods in general.

Electrochemistry↗
PubMed1995

Determinants of late allograft nephrectomy.

When loss of graft function occurs more than six months after transplantation, allograft nephrectomy is not routinely performed at the time of graft failure. It is usually performed only on those patients who subsequently develop specific complications. However, little is known about the characteristics that make patients more likely to require allograft nephrectomy. The purpose of our study was to identify risk factors for the subsequent need for allograft nephrectomy in patients with graft failure occurring more than 6 months after transplantation. Forty-one patients were studied. Inclusion criteria were: loss of graft function > or = 6 months after transplantation, resumption of dialysis and initiation of weaning from immunosuppression. Thirty patients were treated with cyclosporine + prednisone +/- azathioprine and 11 with azathioprine + prednisone. Mean follow-up time was 17.8 months, ranging from 6 months to 6.1 years. Recipient age, sex and race, original renal disease, donor, donor source (cadaveric vs living related), HLA compatibility, levels of panel reactive antibodies, occurrence of initial delayed graft function, causes of graft failure and tapering of immunosuppression were similar in patients with and without allograft nephrectomy. Using univariate analysis, allograft nephrectomy was found to be significantly more frequent in patients with a history of 2 or more episodes of acute rejection than in patients with no rejection episode: 83% vs 30% (p = 0.03). In addition, allograft nephrectomy was found to be significantly more frequent if the immunosuppressive regimen included cyclosporine (62% vs 27.3%; p = 0.04). Using multivariate analysis however, the number of previous episodes of rejection was found to be the only significant predictor for allograft nephrectomy. None of the other variables considered in the multivariate analysis, including the type of immunosuppressive therapy, was identified as a significant predictor for the need to perform allograft nephrectomy. In summary, the need for late allograft nephrectomy was correlated with the number of previous episodes of acute rejection. Patients with a history of numerous rejection episodes should thus be considered more likely to require allograft nephrectomy once immunosuppression is withdrawn. Possible interventions to reduce or prevent the need for nephrectomy include more gradual tapering of immunosuppression at the time of graft failure or indefinite low-dose immunosuppressive therapy.

Adult↗
PubMed1994

Phosphatidylinositol-anchored molecules and inducible lipopolysaccharide binding sites of human and mouse bone marrow cells.

We have previously established that lipopolysaccharide (LPS) induces the expression of new specific LPS-binding sites (LpsR) in mouse bone marrow cells (BMC). We now show that exposure of human BMC to LPS elicits the production of both CD14 molecules (detectable with monoclonal antibody My4) and LpsR (detectable with fluorescein isothiocyanate-LPS). Pretreatment of stimulated human BMC with My4 inhibited the binding of fluorescein isothiocyanate-LPS. The stimulation of human BMC, but not mouse BMC, required the presence of serum. Other characteristics of mouse and human BMC examined were very similar. Their inducible LpsR interacted with the lipid moieties of LPS and Leishmania donovani lipophosphoglycan and with a soluble preparation of peptidoglycan. Moreover, mouse and human LpsR were susceptible to treatment with a phosphatidylinositol-specific phospholipase C (PI-PLC), thus suggesting that both are PI-anchored CD14 molecules. Neither LpsR appeared able to interact with a synthetic LPS antagonist (compound PPDm2) structurally related to the lipid region of LPS. However, PPDm2 blocked LPS-induced expression of LpsR in both BMC. Furthermore, in both species, pretreatment of BMC with PI-PLC did not prevent the cells from expressing LpsR in response to LPS. The results support the hypothesis that the elicited LpsR of mouse and human BMC is an inducible form of CD14, whereas the putative "signaling LPS receptor" of these cells is not CD14 or any other PI-anchored molecule.

Acute-Phase Proteins↗
PubMed1993

Lipopolysaccharide and tumor necrosis factor-alpha induce lipopolysaccharide receptor expression on bone marrow cells by different mechanisms.

Two cell types, monocytes/macrophages and neutrophilic granulocytes, play a prominent role in the pathogenic effects of endotoxins during Gram-negative infections. We previously established that nanomolar concentrations of LPS induce the expression of new specific LPS-binding sites (LpsR) in bone marrow granulocytes of LPS-responsive mice. To examine this induction process further, we asked whether it, like other LPS activities, can be mediated by TNF-alpha. We report that exogenous rTNF-alpha can induce LpsR expression in bone marrow cells (BMC) from both LPS-responsive (C3H/HeOU) and LPS-hyporesponsive (C3H/HeJ) mice. In BMC, LPS elicited a down-regulation of the TNF-alpha receptors (TNF-R) without direct binding to TNF-R. On the other hand, taxol, a microtubule stabilizer that has shown LPS mimetic activity in macrophages, was unable to elicit LpsR expression or induce TNF-R down-regulation in BMC. Thus, unlike the LPS-signaling receptor of macrophages, that of BMC is apparently not functionally associated with microtubules. The LPS-induced expression of LpsR was inhibited only partially with an anti-TNF-alpha serum, and with dexamethasone, suggesting that an autocrine activity of endogenous TNF-alpha cannot alone account for the LPS effect. Comparative analyses also indicated that dexamethasone inhibited the LPS-induced increase of LpsR, but enhanced the number of TNF-induced LpsR+ BMC. Furthermore, the synthetic lipid PPDm2 (a 1,4-bisphosphorylated and N,N-diacylated derivative of 2,3-diamino-2,3-dideoxy-D-glucose) inhibited LPS-induced, but not TNF-induced, expression of LpsR. These data show that in BMC, LPS and TNF-alpha induce LpsR expression by different mechanisms.

Animals↗
PubMed1993

Endotoxin-induced expression of endotoxin binding sites on murine bone marrow cells.

A variety of binding sites for endotoxin (LPS) have been identified on leukocytes. However, the sequence of expression of these receptors, and their interrelations, are poorly understood. In this report, we show that in LPS-responsive hosts, interaction of nanomolar concentrations of LPS with bone marrow cells induces the expression of new specific LPS-binding sites. Cells from LPS-nonresponsive (C3H/HeJ) mice do not express these receptors after LPS treatment. Experimental differences in the conditions allowing the induction and the detection of these binding sites (influence of Leishmania lipophosphoglycan, role of serum), support the hypothesis that interaction of LPS with primary receptors on bone marrow cells triggers the expression of secondary LPS receptors, and that the two types of receptors have distinct fine specificities.

Animals↗
PubMed1993

Differential effects of a monoclonal antibody recognizing 3-deoxy-D-manno-2-octulosonic acid on endotoxin-induced activation of pre-B cells and macrophages.

A monoclonal antibody (E1), which reacts with the 3-deoxy-D-manno-2-octulosonic acid region of lipopolysaccharides belonging to the rough Re chemotype (LPS-Re) was used to analyze in vivo and in vitro effects of endotoxin. E1 inhibited LPS-Re-induced activation of Z0Z/3 pre-B cells (expression of surface immunoglobulins) and mature splenic B cells (DNA synthesis) and blocked the binding of biotin-labeled LPS-Re to mouse peritoneal macrophages. However, other effects elicited by LPS-Re, such as the production of interleukin 6 and tumor necrosis factor alpha by macrophages, and the acute lethality in galactosamine-sensitized mice, were not inhibited by E1. The results suggest that the specificity of the LPS receptor which triggers the production of cytokines in macrophages is different from that of the LPS receptor which induces activation of pre-B and B lymphocytes, and is also different from that of the major LPS-binding sites of macrophages. This is consistent with the hypothesis that the large majority of LPS-binding sites of macrophages do not trigger cytokine production, and that the small number of "signaling LPS receptors" of macrophages have fine specificities which are different from those of B cells.

Animals↗
PubMed1993

Nontoxic forms of lipopolysaccharides, poorly cleared in vivo, are present in variable amounts in phenol-extracted endotoxins.

A radiolabeled lipopolysaccharide (LPS) from Salmonella choleraesuis was as toxic for galactosamine-treated mice as the unlabeled preparation. After intravenous injection, a large proportion (37%) of the labeled material remained in the circulation. This circulating form of LPS was not toxic, even when reextracted with phenol, and remained in the circulation when reinjected into mice. Furthermore, toxic and nontoxic constituents were directly isolated in vitro by hydrophobic gel chromatography of the radiolabeled LPS preparation. The nontoxic form, of lower molecular weight according to its electrophoretic migration, did not react with an anti-lipid A monoclonal antibody but reacted with polymyxin B and was as active on pre-B and B cells as the unfractionated LPS preparation. The results of this study suggest that some LPS constituents of phenol-extracted endotoxin preparations are not toxic for galactosamine-sensitized mice and are not trapped by appropriate LPS-binding sites in the liver as efficiently as are the toxic forms.

Animals↗
PubMed1993

A synthetic analog of the 3-deoxy-D-manno-2-octulosonic acid disaccharide moiety of rough-type endotoxins does not bind to mouse peritoneal macrophages and human monocytes.

Strong evidence supports the concept that lipid A is the main biologically active region of endotoxins and is recognized by specific binding sites of different cell types. However, receptors for carbohydrates are also present on mononuclear phagocytes, and it has been suggested that one of these lectin-like proteins may be specific for the 3-deoxy-D-manno-2-octolosonic acid (Kdo) residues of endotoxins. To reexamine this hypothesis, we prepared a 125I-labeled conjugate consisting of a synthetic Kdo-2,4-Kdo disaccharide covalently linked to bovine serum albumin (125I-Kdo2-BSA). The Kdo disaccharide residues of this radiolabeled conjugate were fully accessible to a monoclonal antibody which reacts specifically with this epitope. However, 125I-Kdo2-BSA did not exhibit any detectable specific binding on thioglycolate-elicited mouse peritoneal macrophages or on human monocytes. Furthermore, the specific binding of biotin-labeled lipopolysaccharide derivatives to mouse macrophages and human monocytes was not inhibited by a soluble synthetic Kdo-2,4-Kdo-polyacrylamide copolymer or by a synthetic glycolipid consisting of an alpha-Kdo residue glycosidically linked to O-6 of allyl-4-O-phosphoryl-N-3-hydroxytetradecanoyl-beta-D-glucosaminide. These results indicate that binding sites specific for Kdo are not present (or not accessible) on the surface of mouse macrophages and human monocytes.

Animals↗