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Biomedical subjects

M Verdier

Publications and source records attributed to M Verdier.

At least 19 recordsLinked to original sources
PubMed2006

Activation of Bad trafficking is involved in the BCR-mediated apoptosis of immature B cells.

The activity of Bad, a pro-apoptotic protein, is regulated by reversible phosphorylation. Moreover, sequestration of Bad within subcellular compartments may be a new mechanism of apoptosis regulation. In this study, we report that Bad interacts with 14-3-3 protein in WEHI-231 immature B cells. This association is disrupted following BCR stimulation in correlation with Bad translocation to mitochondria and apoptosis. Confocal microscopy was further used to examine the co-localization of Bad with lipid rafts in WEHI-231 and murineex vivoB cells. Bad was found colocalized to lipid rafts in freshly isolated mature B lymphocytes, in contrast to immature cells. Finally, co-immunoprecipitation experiments performed on WEHI-231 B cells revealed that PP1alpha interacts with Bcl-2 and Bad, and dissociation of the complex was found correlated with appearance of apoptosis. Bcl-2 seemed to be required to assemble the complex which may regulate Bad phosphorylation status and consequently cell survival. Collectively, present data outline the role of Bad trafficking in the BCR-mediated apoptosis and suggest that differences in intracellular Bad trafficking may be involved in the differential outcome of BCR signaling.

14-3-3 Proteins↗
PubMed2006

11beta-Hydroxysteroid dehydrogenase type 1-driven cortisone reactivation regulates plasminogen activator inhibitor type 1 in adipose tissue of obese women.

BACKGROUND: Plasminogen activator inhibitor type 1 (PAI-1) is the main inhibitor of the fibrinolytic system and contributes to an increased risk of atherothrombosis in insulin-resistant obese patients. In adipose tissue, we have shown that PAI-1 is synthesized mainly in the visceral stromal compartment and is positively regulated by glucocorticoids. We have demonstrated that adipose tissue expression of 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD-1), an enzyme that catalyzes the conversion of inactive cortisone to active cortisol, is exaggerated in obese patients. OBJECTIVES: We hypothesized that increased action of 11beta-HSD-1 in adipose tissue of obese subjects may contribute to PAI-1 overproduction. PATIENTS AND METHODS: Using in situ hybridization, we studied the expression of the mRNAs coding for PAI-1 and 11beta-HSD-1 in the stromal compartment of visceral adipose tissue obtained from obese women. The regulation of PAI-1 secretion from in vitro incubated tissue explants was also investigated. RESULTS: Regression analysis showed a significant positive linear relationship between PAI-1 and 11beta-HSD-1 mRNAs expression. In vitro incubation of adipose tissue explants demonstrated that cortisone stimulated PAI-1 gene expression and secretion, and that these effects were inhibited by co-incubation with the 11beta-HSD inhibitor, glycyrrhetinic acid. CONCLUSIONS: Our data demonstrate that 11beta-HSD-1-driven cortisone reactivation regulates adipose PAI-1 synthesis and secretion. They suggest that the increased PAI-1 synthesis and secretion observed in obese patients can be also related, at least in part, to an increased local conversion of cortisone to cortisol. Therefore, local cortisol metabolism in adipose tissue may be involved in increasing the risk of cardiovascular disease in obese subjects.

11-beta-Hydroxysteroid Dehydrogenase Type 1↗
PubMed2005

Effect of microstructure on the mechanical properties of Haversian cortical bone.

The mechanical properties of cortical bone have been extensively studied at the macrostructural scale. However, knowledge of the macroscopic mechanical properties is not sufficient to predict local phenomena, such as damage or bone remodeling, both of which are dependent on local mechanical behavior. The objective of this study is to quantify the mechanical properties of cortical bone at several length scales, with emphasis on the microstructure of Haversian systems. Samples of mature bovine cortical bone, with a Haversian microstructure, were obtained from the posterior area of the mid-femoral diaphysis. A nanoindentation technique was used to measure the local Young's modulus. The distribution of the bone mineral content was obtained by backscattered electron imaging using a scanning electron microscope. A novel compression device employing microextensometry techniques was developed to quantify local strains. Digital image correlation was performed on the microstructure imaged by optical microscopy during compression tests. This study demonstrated that the local Young's modulus and strain were heterogeneous at the scale of an osteon. For both properties, the ratio between the maximum and minimum values was approximately two. The local Young's modulus and bone-mineral content were reasonably correlated (r2 = 0.75; P < 0.0001), but this was not the case for the distribution of local strains versus bone mineral content (r2 = 0.395; P < 0.0001). Hence, local strains cannot be described simply in terms of the bone mineral content, as the Haversian canal and osteonal microstructure have a major influence on these properties. In conclusion, the microstructure must be considered in evaluating the local strain and stress fields of cortical bone.

Animals↗
PubMed2005

Successful use of oviduct epithelial cell coculture for in vitro production of viable red deer (Cervus elaphus) embryos.

Techniques for in vitro production (IVP) of viable embryos have been thoroughly developed in several domestic species in view to improve breeding efficiency. When applied to wild life, these techniques may also help the maintenance of biodiversity through amplification of sparse animals offspring and facilitation of genetic material exchange. During the successive steps of IVP, i.e. oocyte in vitro maturation (IVM), fertilization (IVF) and early embryo development (IVD) to the blastocyst stage, gametes and embryos are faced with unusual environment, including oxidative stress, known to be detrimental to their survival. In the present study, starting from methods developed in domestic species, we have adapted IVP to produce viable red deer embryos. In a first experiment, cumulus cells were removed from in vitro matured oocytes either before or after IVF. The presence of cumulus cells during IVF did not affect final cleavage or development rates. In a second experiment, in vitro matured oocytes were fertilized in the presence of cumulus cells and cultured in SOFaaBSA medium alone or in the presence of ovine oviduct epithelial cell (oOEC) monolayer. Whereas, oviduct cells did not improve the cleavage rate, they significantly increased the rate of embryos reaching the blastocyst stage (from 3 to 25% of total oocytes). Ten blastocysts from oOEC coculture were transferred after freezing and thawing to five recipient hinds and gave rise to three pregnancies. The three pregnant hinds gave birth to three live and normal calves.

Animals↗
PubMed2004

Imaging a dense nanodot assembly by phase retrieval from TEM images.

Phase retrieval is a classical inverse problem in many fields dealing with waves that is becoming of increasing interest in transmission electron microscopy (TEM). A non-interferometric approach is here applied to TEM images. Phase retrieval possibilities given by the transport intensity equation are compared to the ones deriving from the weak phase object approximation. In the limit of small angles, both methods lead to a similar equation between the phase and a set of defocus images. This equation can be solved by an image processing equivalent to using a specific filter in Fourier space. This processing leads to phase images with a spatial resolution here essentially limited by the defocus amount between images. A dense assembly of silicon nanodots is used as a model case to illustrate the interest of this approximate phase retrieval method which can be carried out on standard equipment. The dot heights estimated using the phase images are found to be in good agreement with ones measured by atomic force microscopy. Since image noise and large defocus values may strongly affect the solution given by the approximate method, an iterative phase retrieval method is also used as a test for working conditions.

Fourier Analysis↗
PubMed2003

Changes in bad phosphorylation are correlated with BCR-induced apoptosis of WEHI-231 immature B cells.

Signaling through the B cell antigen receptor (BCR) is a key determinant in the regulation of B cell physiology. Depending on additional factors, such as microenvironment and developmental stage, ligation of the BCR can trigger B lymphocyte activation, proliferation, or apoptosis. The regulatory mechanisms determining B cell apoptosis and survival are not completely known. Using the murine B lymphoma cell line WEHI-231 as a model system, we investigated the role of Bad phosphorylation, a pro-apoptotic member of the Bcl-2 family, in anti-IgM mediated apoptosis. For apoptotic analysis we focused in particular on the mitochondrial potential (deltapsi(m)) collapse which has been reported as a rate-limiting step in the BCR-induced cell death of immature B lymphocytes. Bad phosphorylation at serine 112, 136 and 155 was found in WEHI-231 cell control cultures and its hypophosphorylation on the three sites correlated with the appearance of apoptosis when cross-linking surface IgM. Furthermore, treatment of cells with specific PK inhibitors known to be involved in serine phosphorylation of Bad (LY294002 for PI3K and H-89 for PKA) mimiced or enhanced BCR-induced cell death. These results strongly suggest that regulation of Bad phosphorylation plays an active role in mediating anti-IgM-induced apoptosis of immature B cells.

14-3-3 Proteins↗
PubMed2001

Two new alleles of the RHCE gene in Black individuals: the RHce allele ceMO and the RHcE allele cEMI.

Six unrelated individuals of Afro-Caribbean origin, whose red cells have a marked reduction of the Rhe antigen expression, have been identified. All exhibited the same serological profile with anti-e monoclonal antibodies and lacked expression of the high frequency e-related antigen hrS. Transcripts and genomic analysis showed that these phenotypes resulted from the presence of two new RHCE alleles, ceMO and cEMI. The ceMO allele corresponded to a RHce gene carrying a G667T mutation (exon 5) and was detected at the homozygous state in sample 1 and at the heterozygous state in samples 2-6. The G667T mutation resulted in a Val223Phe substitution on the Rhce polypeptide, in close proximity to Ala226 (e-antigen polymorphism), which might account for the altered expression of e. The ceMO allele is also associated with the lack of expression of the hrS antigen. The absence of the hrS antigen expression may have implications in transfusion as hrS-negative individuals may develop clinically significant antibodies. The cEMI allele corresponded to a silent RHE allele carrying a nine nucleotide deletion within exon 3 and was detected at the heterozygous state in sample 2. This deletion resulted in a shortened polypeptide of 414 residues (instead of 417) that was absent (or severely reduced) at the red cell surface, as the E antigen was undetectable using serology and Western blot analysis with anti-E reagents. In DNA-based polymerase chain reaction genotyping for RHE determination, the cEMI allele provided a false positive result as the cells carrying this allele are serologically phenotyped as E-negative. The incidence of this allele in the Black population is unknown but, as shown already for D genotyping, one must exercise caution when genotyping is performed to detect the e/E polymorphism.

Africa↗
PubMed2001

Sequential gene expression of P-glycoprotein (P-gp), multidrug resistance-associated protein (MRP) and lung resistance protein: functional activity of P-gp and MRP present in the doxorubicin-resistant human K562 cell lines.

Previous studies have reported that P-glycoprotein (P-gp), a transmembrane efflux pump involved in multidrug resistance (MDR), was overexpressed in the doxorubicin (Dox)-resistant human erythroleukemia cell line K562. Nevertheless, several results suggested that P-gp was not the only mechanism involved in these resistant cells. Sequential co-expression of other MDR-associated proteins was sometimes reported, as MDR-associated protein (MRP) and lung resistance protein (LRP), in different MDR cell lines. Thus, mRNA expression and stability of P-gp, MRP and LRP were analyzed, while their corresponding protein levels were quantified in correlation with functional assay, in the K562 cell line and two Dox-resistant variants (K562/R). Their P-gp content was in accordance with their degree of resistance, but not as much in the level of mRNA expression, suggesting a post-transcriptional regulation. On the other hand, MRP could play a minor role in MDR because of an unchanged expression in K562/R sublines. A surprising progressive disappearance of LRP in both resistant cells suggested that the original mechanism of drug redistribution may be operative, involving a negative role for LRP.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2000

Optimization of cell permeabilization for multiparametric flow cytometric analysis with lectin staining.

BACKGROUND: This study was undertaken in mice to develop a reproducible procedure of cell permeabilization, allowing intracellular protein staining by immunofluorescence (i.e., Bcl-2) without losing surface labeling especially for lectins (i.e., B220 and peanut agglutinin [PNA]). This article reports results obtained with different permeabilization protocols. METHODS: Lymphoid cells were extracted and prepared from Peyer's patches and spleen. After surface labeling using anti-B220-Cy-chrome and PNA-biotin/streptavidin-phycoerythrin, we comparatively tested three permeabilization protocols: saponin 0.3%, methanol 70%, and the commercial kit Dako Intrastain. Final Bcl-2 staining was performed and cells were analyzed by flow cytometry. RESULTS: With 0.3% saponin as the permeabilization reagent, a significant loss of lectin labeling was observed when comparing mono PNA and triple (i.e. , B220-PNA-Bcl-2) staining (74.8% and 22.5% positive cells, respectively). Quality of PNA staining was conserved with Intrastain when comparing multiparametric versus monoparametric stainings (82. 4% of positive cells versus 78.3%, respectively). Intrastain preserved scatter characteristics (69.9% of total cells in the lymphocyte gate with Intrastain versus 13.7% with saponin 0.3% and 20.9% with methanol 70%). This protocol has been used for a preliminary multiparametric analysis in order to quantify Bcl-2 expression in PNA/B220-positive cells. CONCLUSION: This protocol may be useful to assess simultaneously lectin cell surface labeling and intracellular target staining.

Animals↗
PubMed2000

Plasminogen activator inhibitor 1, transforming growth factor-beta1, and BMI are closely associated in human adipose tissue during morbid obesity.

In adipose tissue from both obese mice and humans, plasminogen activator inhibitor 1 (PAI-1) expression has been reported to be upregulated to levels of increased plasma PAI-1. This elevated expression has been shown to be partly controlled by tumor necrosis factor (TNF)-alpha in mice. In humans, increased PAI-1 expression is associated with insulin resistance characterized by visceral fat accumulation. Therefore, the aim of this study was to investigate the expression pattern of PAI-1 and TNF-alpha (antigen and mRNA) in visceral human adipose fat in comparison with subcutaneous (SC) fat. Because transforming growth factor (TGF)-beta1 is a potent inducer of PAI-1 synthesis and has been shown to influence adipocyte metabolism, this work was extended to TGF-beta1 quantification. A total of 32 obese individuals (BMI 42 +/- 6.8 kg/m2) were investigated. Freshly collected visceral adipose tissue did not exhibit a higher content of PAI-1 or TGF-beta1 than did SC tissue. Although most of the TNF-alpha values were at the detection limit of the methods, TNF-alpha antigen was 3-fold higher and TNF-alpha mRNA was 1.2-fold higher in visceral fat. The levels of tissue TGF-beta1 antigen correlated well with those of PAI-1 antigen, regardless of the fat depot studied (SC tissue: n = 21, r = 0.72, P = 0.0006; visceral tissue: n = 20, r = 0.49, P < 0.03), and they were both significantly associated with BMI. Conversely, no relationship was observed between the levels of TNF-alpha and PAI-1 or TNF-alpha and BMI. Tissue PAI-1 levels were also significantly correlated with those of circulating PAI-1. These results describe, in severe obesity, a proportional increase in tissue PAI-1 and TGF-beta1 in visceral and SC tissues. This increased PAI-1 expression could be the result of tissue cytokine disturbances, such as elevated TGF-beta1 expression.

Adipose Tissue↗
PubMed1999

PAI-1 produced ex vivo by human adipose tissue is relevant to PAI-1 blood level.

Human adipose tissue has been shown to produce plasminogen activator inhibitor type 1 (PAI-1). However, the importance of adipose tissue in the regulation of the PAI-1 plasma level is not known. The aim of this study was to investigate the relation between the production of PAI-1 by adipose tissue, plasma PAI-1 level, and variables related to the insulin resistance state. The link between the production of PAI-1 inducers such as tumor necrosis factor-alpha and transforming growth factor-beta and the production of PAI-1 by adipose tissue was also evaluated. Blood samples were obtained as soon as possible to the induction of anesthesia from 30 patients undergoing elective abdominoplasty. PAI-1 antigen levels measured in conditioned media after a 19-hour incubation period of adipose tissue explants were significantly correlated with plasma PAI-1 antigen levels (r=0.54, P=0.004) and with systemic lipid parameters such as triglycerides and high density lipoprotein cholesterol (r=0. 46, P=0.014; r=-0.50, P=0.01, respectively) but not with insulinemia and body mass index. PAI-1 production by adipose tissue was correlated with those of TNF-alpha (r=0.5, P=0.01) and TGF-beta (r=0. 53, P=0.007). These results emphasize the role of adipose tissue in determining plasma levels of PAI-1, with a local contribution of TNF-alpha and TGF-beta in PAI-1 production by adipose tissue.

Adipose Tissue↗
PubMed1999

Glucocorticoids and insulin promote plasminogen activator inhibitor 1 production by human adipose tissue.

Plasminogen activator inhibitor 1 (PAI-1) is likely to play a role in vascular disease, primarily in subjects with android obesity. It has been demonstrated that PAI-1 is overexpressed in adipose tissue from obese subjects and that visceral adipose tissue produced more PAI-1 than subcutaneous fat. In the present study, the effect of insulin and glucocorticoids, which are key mediators of adipose tissue metabolism, was examined in relation to PAI-1 synthesis by human adipose tissue explants (HAT), collagenase isolated human adipocytes (IHA), cultured human stromal cells (cSC), and differentiated adipocytes from the murine clonal cell line 3T3-F442A. A significant increase in PAI-1 antigen release (1.5-fold) from HAT was detectable after 16 h of treatment with insulin concentrations of at least 10(-8) mol/l. This was associated with a PAI-1 mRNA increase. Concomitant addition of insulin (10(-8) mol/l) to forskolin (5 x 10(-5) mol/l) reversed the decrease in PAI-1 antigen caused by forskolin alone. No effect on PAI-1 antigen was observed when insulin was incubated with IHA or cSC. 3T3 F442A cells were sensitive to insulin with a four- and twofold increase in PAI-1 antigen and mRNA levels, respectively, after 16 h of stimulation with 10(-8) mol/l. Dexamethasone (DXM) significantly enhanced PAI-1 antigen and mRNA expression by HAT (1.5- and 2.5-fold increase, respectively) at concentrations of at least 10(-8) mol/l. A higher stimulation was observed with IHA (sevenfold increase) and with the differentiated 3T3 F442 cell line. Cortisol was found to be less potent than DXM. No effect was observed when glucocorticoids were incubated with cSC. Coincubation of HAT with insulin (10(-7) mol/l) and DXM (10(-7) mol/l) led to an additive effect on PAI-1 synthesis. These results support the hypothesis that PAI-1 expression in human adipose tissue is controlled by insulin and glucocorticoids and may help to explain the increase in plasma PAI-1 levels observed in patients with android obesity.

3T3 Cells↗
PubMed1998

Intrafamilial clustering and 4-year follow-up of asymptomatic human T-cell leukaemia virus type I (HTLV-I) infection in Benin (West Africa).

BACKGROUND: Few data exist concerning familial human T-cell leukaemia virus type I (HTLV-I) carrier states and transmission in African countries. Two previous surveys performed in Benin in 1989 and 1990 using a three-level cluster sampling method allowed us to identify HTLV-I positive subjects. The evolution of HTLV-I within the families of these subjects is described over a 4-year period, 1991-1995. METHODS: Since 1991, 37 HTLV-I seropositive subjects, six subjects with indeterminate Western-Blot pattern, and their relatives have been followed up once a year clinically and biologically. RESULTS: Twenty-three mothers in the study group gave birth to 27 children between 1991 and 1995. Among the 13 infants born to the 12 seropositive mothers, two seroconverted before their second birthday. One adult woman whose husband was seropositive developed seropositivity 4 years after marriage. In March 1992, a family case-control study (proband study) was conducted. A seroprevalence of 27.5% was found among 138 relatives of 32 infected subjects and 1.4% among 142 relatives of 32 control subjects. CONCLUSIONS: There is clearly an intrafamilial clustering of HTLV-I in Benin. The annual incidence density of HTLV-I in this cohort is estimated at 6 per thousand.

Adolescent↗
PubMed1996

Seroepidemiological study of retroviruses (HTLV-I/II, HIV-1, HIV-2) in the Department of Atacora, northern Benin.

A seroepidemiological survey to determine the prevalence of retrovirus infection (HTLV-I/II, HIV-1, HIV-2) by representative sampling of the general population in the Department of Atacora in north-western Benin is reported. The seroprevalence rate of HTLV-I in this sample was at 1.86% (95% CI 1.20-2.52%). This is in agreement with prevalence rates reported from neighbouring countries of the sub-region. No sera were found positive for HTLV-II. Seropositivity to HIV-1 was 0.3%; HIV-2 seropositivity was not encountered.

Adolescent↗
PubMed1995

Stedim 6 and Clearflex, two new multilayer materials for infusion containers. Comparative study of their compatibility with five drugs versus glass flasks and polyvinyl chloride bags.

Stedim 6 and Clearflex, two new polyethylene-lined materials for infusion bags, were studied for their compatibility with disodium clodronate, chlorpromazine and maprotiline hydrochlorides, diazepam, and clorazepate dipotassium salt, comparatively with borosilicate glass flasks and polyvinyl chloride bags. Diazepam, the only drug to exhibit a marked sorption in PVC bags (the loss reached 25% of the initial concentration after a contact duration of 72 h), showed lower sorption in Stedim 6 bags (loss about 11% under the same conditions) and none in Clearflex bags. No significant difference was observed between the infusion solutions used as vehicles of the drugs (5% dextrose and 0.9% sodium chloride isotonic solutions). The results are discussed in terms of lipophilicity of the drugs and crystallinity of the polymers.

Adsorption↗
PubMed1995

[Intra-family transmission of human T-cell lymphotropic virus-1 (HTLV-1) in a cohort of HTLV-1-positive patients (Bénin)].

We are reporting the results of a familial study carried out in Benin in March 1994 within a cohort of HTLV-1 positive subjects. This study aims at appraising the different modes of intra-familial transmission of this retrovirus. The study has included 212 persons: 33 seropositive subjects (identified during two previous seroprevalence surveys and followed up since 1991), and 179 members of their families. Blood specimens have been taken from each of these subjects. Sera have been screened for HTLV-1 antibodies by ELISA test and positive results confirmed by Western blot test. Out of 18 children born during the follow up period, 2 cases of seroconversion have been observed. One case of seroconversion has also been noted among the 17 couples in which either of the spouses is seropositive. Among the 136 children of the cohort: 17.8% of them are HTLV-1 positive when both parents are seropositive 26.1% if the mother only is seropositive and 0% if the father only is seropositive. The incidence rate in this cohort is estimated to be 0.43%. These data relative to the intra-familial transmission of HTLV-1 in Benin appear to be consistent with those previously reported in Japan.

Benin↗