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Biomedical subjects

A Lefèvre

Publications and source records attributed to A Lefèvre.

At least 19 recordsLinked to original sources
PubMed2003

Role of the interaction matrix in mean-field spin glass models.

Mean-field models of two-spin Ising spin glasses with interaction matrices taken from ensembles that are invariant under O(N) transformations are studied. A general study shows that the nature of the spin glass transition can be deduced from the eigenvalue spectrum of the interaction matrix. A simple replica approach is derived to carry out the average over the O(N) disorder. The analytic results are confirmed by the extensive Monte Carlo simulations for large system sizes and by the exact enumeration for small system sizes.

Journal Article↗
PubMed2002

Multiple parameter cytotoxicity index on dental alloys and pure metals.

Palladium (Pd) is a metal frequently used for dental alloys. In order to elucidate controversial options about Pd concerning its biological performances, our study consists in the evaluation of commercial and experimental PFM and C&B precious and semi-precious dental alloys. This investigation was also designated to the establishment of a cytotoxicity index (CI) such as it was described for hemocompatibility testing. The following materials were tested: 36 commercial alloys (Au-, Pd- and Ag-base), 14 experimental alloys (Pd-base established by an experience plan) and pure metals (Ag, Au, Cu, Ni, Cr, In, Sn, Pt, Ti, Zn). The cells culture experiments were carried out with epithelial L132 cells and NIH 3T3 fibroblasts. In vitro cell viability tests show that Pt, Sn, In, Ti, Au and Pd have no cytotoxic effect; Cr, Cu and Ag are toxic, Ni, Zn, and Co are highly toxic. An identical ranking was found with the inflammatory and proliferation tests. Toxic and highly toxic metals induced slight or strong prosthetic dental restoration morphological alterations after 3-days cultures and mostly cell death after 6-days cultures. These effects are dependent on the leakage of the element into the culture medium as revealed by ICP. The addition of Au gives benefit to Pd-Ag alloys, but does not produce any major effect on Pd-Cu alloys. This qualitative ranking can quantitatively be confirmed by cytocompatibility testing after application of a CI.

Animals↗
PubMed2002

In vitro studies of human and rat osteoclast activity on hydroxyapatite, beta-tricalcium phosphate, calcium carbonate.

Investigations on the ceramic degradation caused by osteoclasts are designed to assess osteoclast-ceramic interactions and to determine which ceramics are more suitable for use as bone substitute. This study investigated the resorptive activity of osteoclasts on ceramics presenting different solubility rates. Osteoclasts isolated from new-born rat and from human giant cell tumour were cultured on different bioceramics: hydroxyapatite (HA), beta-tricalcium phosphate (TCP) and calcium carbonate (calcite). Cytoskeletal was revealed by actin labelling and ceramic surfaces were observed by scanning electron microscopy (SEM). On all materials, the distribution of actin in typical ring was revealed. SEM examinations showed a clear difference in the shape and the depth of resorption lacunae on different ceramics. On pure HA, a superficial attack, clearly visible but very little extended. Numerous resorption lacunae, deep and well-delimited were observed on pure beta-TCP, but attacks less punctually were detected too. On pure calcite, an attack with form of spikes, very widespread but superficial was revealed. Degradation measurements revealed a significant increase of P release from the phosphocalcic ceramics and of Ca from all ceramics in the presence of osteoclasts. The both cell models found these characteristics, the rat osteoclasts were also an excellent model to study the ceramic resorption.

Adsorption↗
PubMed2001

Steady state behavior of mechanically perturbed spin glasses and ferromagnets.

A zero temperature dynamics of Ising spin glasses and ferromagnets on random graphs of finite connectivity is considered. Like granular media, these systems have an extensive entropy of metastable states. We consider the problem of what energy a randomly prepared spin system falls to before becoming stuck in a metastable state. We then introduce a tapping mechanism, analogous to that found in real experiments on granular media. This tapping, corresponding to flipping each spin with probability p simultaneously, leads to a stationary regime with a steady state energy E(p). We explicitly solve this problem for the one-dimensional ferromagnet and the +/-J spin glass, and carry out extensive numerical simulations for spin systems of higher connectivity. In addition our simulations on the ferromagnetic systems reveal a first order transition, whereas the usual thermodynamic transition on these graphs is second order.

Journal Article↗
PubMed2001

Tapping spin glasses and ferromagnets on random graphs.

We consider a tapping dynamics, analogous to that in experiments on granular media, on spin glasses and ferromagnets on random thin graphs. Between taps, zero temperature single spin flip dynamics takes the system to a metastable state. Tapping corresponds to flipping simultaneously any spin with probability p. This dynamics leads to a stationary regime with a steady state energy E(p). We analytically solve this dynamics for the one-dimensional ferromagnet and +/-J spin glass. Numerical simulations for spin glasses and ferromagnets of higher connectivity are carried out; in particular, we find a novel first order transition for the ferromagnetic systems.

Journal Article↗
PubMed2001

Trans iliac-sacral-iliac bar stabilisation to treat bilateral lesions of the sacro-iliac joint or sacrum: anatomical considerations and clinical experience.

The use of trans iliac-sacral-iliac bars is an alternative to sacro-iliac screws in the treatment of bilateral lesions of the posterior pelvic ring, and the same biomechanical principles can be applied. Of 20 patients, ten men and ten women, a CT-scan of the pelvis was performed to study the individual and common safe area at the level of S1 and S2. The location and maximal diameter of the individual safe area were studied using a computer-navigation system, displaying images in sagittal, coronal and axial anatomic planes together with a 3-D reconstruction. The common safe area was studied using three points: upper- (UA) and lower anterior corner (DA) of S1 and S2, and the centre of the safe area. It would have been possible to place an iliac-sacral-iliac bar (5 mm or more) in S1 and S2 in all the men, but in the women a bar could only have been inserted in only five in S1 and eight in S2. A statistically significant difference between men and women was found at S1 (P=0.033) but not at S2 (P=0.211). No significant correlation was found between the diameter of the safe area at both levels in men and women and age, height, and weight. Furthermore, no common safe area of 5 mm or more was measured at the same levels.Four patients were treated using trans iliac-sacral-iliac bars. Three were placed under fluoroscopic control in combination with a frame, and in one patient an image-guided system was used. A postoperative CT confirmed the correct position of the bars in each patient. The complexity and individual variability of the sacrum makes complex preoperative planning of the iliac-sacral-iliac path mandatory.

Adult↗
PubMed2001

In situ measurement and statistical modelling of Escherichia coli decay in small rivers.

Numerous studies have been carried out on the environmental factors associated with the decay of faecal bacteria in open (fresh or marine) waters. The present study aimed at understanding the fate of bacteria in small streams (flow <20 m3 s(-1)) for which there is a lack of knowledge. An original in situ protocol was developed for measuring the die-off of Escherichia coli (E. coli) from wastewater treatment plants. Based upon 80 values of the decay first-order parameter (K or its inverse T90), collected from five rivers in Normandy (France), a median T90 of 10 h and a minimal T90 of 1.3 h were obtained. K was then modelled as a linear function of variables made up from flow, water temperature and suspended particulate matter (SPM). The set of significant co-variables did not include light indicators. E. coli decay is inversely related to the river flow and it becomes highly significant below 0.3 m3 s(-1). The positive effect of small flows on die-off is increased by water temperature over 15 degrees C, whereas it could be reduced by SPM. The major co-variable of the model (p < 10(-9)) is an empiric composite variable integrating the effect of flow and temperature that explains more than 40% of the variance of K. We interpreted this as an expression of predation by benthic micro-grazers which could be the main cause of E. coli die-off in small streams in temperate countries.

Escherichia coli↗
PubMed2001

Differential gene expression in pre-implantation embryos from mouse oocytes injected with round spermatids or spermatozoa.

BACKGROUND: The use of immature male germ cells to fertilize human oocytes raises several questions. Spermatozoa are normally quiescent, but many genes are transcribed post-meiotically in round spermatids. This creates a novel situation for the oocyte. We have therefore explored the effects on early embryonic development of introducing a fully transcriptionally active round spermatid into the oocyte. METHODS AND RESULTS: Following the micro-injection of spermatozoa or spermatids into mouse oocytes we have analysed the expression, at various times, of six genes in the resulting embryo. Spermatozoa and spermatids produced similar fertilization rates. Hprt was expressed in all embryos at all stages tested. Hsp70.1 was found normally during the 2-cell stage and repressed by the 4-cell stage in embryos from both spermatozoa and round spermatids. However, the amplitude of the signal was greatly reduced in 2-cell embryos from round spermatids. Smcy also showed a disturbed pattern of expression in embryos from round spermatids. Protamine 2, which is normally restricted to the spermatid stage, was expressed following fertilization with round spermatids, but was already repressed at the two pronuclei stage. Ube1Y, which is normally expressed post-meiotically and not during the post-implantatory development, was expressed up to the 2-cell stage in embryos from round spermatids only, and then repressed. Ube1X was also expressed up to the 2-cell stage, but in both embryo types. CONCLUSIONS: We therefore suspect that in embryos fertilized with round spermatids, regulatory mechanisms for inhibiting the inappropriate transcription of male post-meiotically expressed genes are activated following fertilization, permitting the zygotic genome activation to occur, though with some disturbances.

Animals↗
PubMed2001

Cloning and sequencing of SOB3, a human gene coding for a sperm protein homologous to an antimicrobial protein and potentially involved in zona pellucida binding.

We have previously characterized an 18-19 kDa cationic protein, SOB3, that was detected in the epididymis and localized within the acrosome and on the neck region of human spermatozoa. We suggested that it is involved in secondary sperm binding to the zona pellucida. The present study describes its purification to homogeneity by preparative electrophoresis and non-equilibrium pH gradient electrophoresis. Degenerate primers deduced from microsequencing were used to amplify a specific fragment from human epididymal RNA by reverse transcription-polymerase chain reaction (RT-PCR). This 164 bp fragment was extended by 5' and 3'-RACE to obtain the 548 bp full length cDNA. The open reading frame encodes a 170 amino acid protein. SOB3 is a single copy gene. It is 98% identical to prepro-FALL39 and 100% identical to CAP18, two human genes which were initially identified by screening a human bone marrow (lambda)gt11 library, and which encode an antimicrobial protein. Northern blots of human tissues revealed a 1 kb transcript in corpus and cauda epididymis only, while RT-PCR showed presence of the mRNA in the three epididymal regions and also in round spermatids. The above results suggest that SOB3 has two roles in sperm protection and fertilization, depending on its dual origin and final sperm localization.

Amino Acid Sequence↗
PubMed1999

Flow cytometry isolation and reverse transcriptase-polymerase chain reaction characterization of human round spermatids in infertile patients.

Flow cytometry coupled to cell sorting is proposed as a method to isolate round spermatids from testicular biopsies in obstructive azoospermic patients. The cells were separated on the basis of their size and density only. We obtained homogenous populations of alive round spermatids free of lymphocytes and diploid germ cells. The detection of protamine 1 gene (PRM1) and PRM2 expression in the sorted cells proves that these cells are round spermatids. On the contrary, neither the expression of CD3-delta, which is specific to lymphoid cells, nor that of MAGE1, which has been demonstrated in diploid germ cells, could be observed in the round spermatid population even after using a nested polymerase chain reaction (PCR) assay. The flow cytometry procedure failed to isolate round spermatids from ejaculates in non-obstructive azoospermic patients. In > 39 ejaculates tested by reverse transcriptase-PCR, only nine revealed the presence of some round spermatids, as demonstrated by the expression of PRM1. However, these round spermatids did not express PRM2.

Adult↗
PubMed1999

Flow cytometric method to isolate round spermatids from mouse testis.

The purpose of this study was to isolate pure populations of round spermatids from mouse testis by flow cytometry followed by cell sorting. Cell suspensions from mouse testis were enriched in germ cells by centrifugation on a discontinuous Percoll gradient, then analysed using a FACScalibur flow cytometer measuring the cell size and density. A large and well-delimited population of cells (R1) expected to contain round spermatids was observed on the dot plot diagram. Sorted R1 cells were very homogeneous in size (approximately 11 microns) and displayed the characteristic cytological aspect of round spermatids. Spermatid-specific gene expression was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of R1 cells using primers for protamine 2 gene (PRM2) and SP-10. A positive signal for SP-10 was obtained with a single cell using nested primers. The 5.5 kb transcript of c-kit, which is not expressed in spermatids, was not detected by nested RT-PCR, excluding a contamination with spermatogonia. Our results clearly established that flow cytometry followed by cell sorting allows the isolation of a highly homogeneous population of round spermatids from the testis.

Acrosome↗
PubMed1998

SOB3, a human sperm protein involved in zona pellucida binding: physiological and biochemical analysis, purification.

LB5 antibody was selected from a monoclonal antibody (mAb) library directed against human sperm proteins. LB5 mAb detected the corresponding protein SOB3 in the neck region and the flagellum of most live ejaculated sperm while it labelled, in addition, the acrosome of about 10-20% of spermatozoa. The percentage of LB5 acrosome-stained sperm was significantly correlated with the percentages of either spontaneous or A23187-induced acrosome-reacted sperm. While SOB3 could not be detected in the testis, it appeared in spermatozoa from the corpus epididymis segment. LB5 mAb impaired neither sperm motion parameters, acrosomal reaction triggering, nor sperm binding to zona-free hamster oocytes. By contrast, LB5 Fab fragments (200 micrograms/ml) inhibited sperm binding to human zonae pellicidae by 35.7%. If sperm were induced to acrosome react with A23187 prior to LB5 treatment, the inhibitory effect shifted to 59.9%, while no significant effect was observed following A23187 incubation alone. Western blotting of human sperm and cauda epididymis extracts revealed two bands of 18 and 19 kDa. While no cross-reaction was observed with other tested organs, a similar 18-kDa band was revealed in erythocytes and one of 19 kDa in B-lymphocytes. No cross-reactivity could be evidenced in any animal sperm analyzed. SOB3 was first separated in a 17- to 20-kDa preparative electrophoresis fraction and finally purified by isoelectrofocusing according to its pl of 9.8. These results suggest that SOB3 is localized under the outer acrosomal membrane, that it participates in secondary sperm binding to the zona pellucida, and that it shares homologies with the immune system.

Acrosome↗
PubMed1998

[Clinical study and genetic 3q28 locus linkage in 2 Swiss families with Kjer dominant optic atrophy (OPA1)].

METHODS: We examined 20 patients from 2 unrelated Swiss families to describe their clinical phenotype. In addition, a linkage analysis was performed in an attempt to confirm the reported genetic homogeneity of this condition as well as to refine its genomic localization. RESULTS: Two point analysis provided a cumulative LOD-score of 3.03 with marker D3S 2305. The absence of recombination precluded further refinement of the disease interval. CONCLUSIONS: Our data confirm the genetic homogeneity and the extreme variability of expression, occasionally mimicking low tension glaucoma.

Adolescent↗
PubMed1997

Characterization and isolation of SOB2, a human sperm protein with a potential role in oocyte membrane binding.

G12 monoclonal antibody (mAb), one of a library of constructed mAb directed against human sperm proteins, was found by immunoperoxidase staining to label the post-acrosomal and neck regions of fixed human cauda epididymal and ejaculated spermatozoa. Epithelium and fluid of caput epididymis were strongly labelled while there was no staining on testis and efferent ducts. Western lot analysis revealed that G12 antibody reacted with proteins of 17.5, 18 and 19 kDa in human spermatozoa. This pattern seems to be specific for mature human spermatozoa, as it has not been observed either in other human tissues tested, or in spermatozoa from different animals. SOB2, the corresponding protein, was isolated from NP40-extracted human spermatozoa by using preparative electrophoresis, followed by isoelectrofocusing according to its isoelectric point of 6.4 G12 Fab fragments strongly inhibited binding of human spermatozoa to zona-free hamster oocytes (up to 86% inhibition at 200 micrograms/ml). Impairment of binding was dependent on the concentration of purified G12 immunoglobulin (Ig)G1, and significant even at 10 micrograms/ml. There was no inhibitory effect of G12 antibody on sperm motility parameters or triggering of the acrosome reaction and it did not inhibit binding to human zona pellucida. These results indicate that SOB2 is likely to participate in membrane oocyte binding, and my be potential candidate for the development of a contraceptive vaccine.

Animals↗
PubMed1995

FLB1, a human protein of epididymal origin that is involved in the sperm-oocyte recognition process.

CA6 antibody was selected out of a monoclonal antibody library raised against human sperm proteins primarily for its ability to recognize an epididymal antigen and to modify sperm adhesion to zona-free hamster oocytes. In the present study, CA6 was shown to decrease sperm binding to zona-free hamster and human oocytes by 40-92% and 38-48%, respectively. The corresponding protein, which was referred to as FLB1, was found to be secreted by the epididymis and to bind specifically to a human, macaque, and rodent subacrosomal sperm region. Western blotting revealed a molecular mass of 94 kDa in human epididymal extracts and of 100 kDa in human, macaque, mouse, rat, and hamster sperm, suggesting further modifications after its binding to sperm. An equivalent protein was not observed in human liver, ovary, testis, plasma, or epidermis. Two-dimensional electrophoresis showed that FLB1 is formed of two subunits with the same 47-kDa molecular mass and slightly different pI (5.8, 5.9). Microsequencing of the protein revealed a partial homology with human cytokeratins 1 and 10. These results suggest that FLB1 is an epididymis-specific cytokeratin-like protein that is involved in the sperm-oocyte recognition process.

Amino Acid Sequence↗
PubMed1994

Regulation by retinoids of luteinizing hormone/chorionic gonadotropin receptor, cholesterol side-chain cleavage cytochrome P-450, 3 beta-hydroxysteroid dehydrogenase/delta (5-4)-isomerase and 17 alpha-hydroxylase/C17-20 lyase cytochrome P-450 messenger ribonucleic acid levels in the K9 mouse Leydig cell line.

Vitamin A is a potent regulator of testicular function. We have reported that retinol (R) and retinoic acid (RA) induced a down regulation of luteinizing hormone/human chorionic gonadotropin (LH/CG) binding sites in K9 Leydig cells. In the present study we evaluated the effect of R and RA on LH/CG receptors, cholesterol side-chain cleavage cytochrome P-450 (P-450 scc), 17 alpha-hydroxylase/C17-20 lyase (P-450 17 alpha) and 3 beta-hydroxysteroid dehydrogenase (3 beta HSD) mRNA levels in K9 mouse Leydig cells. To validate K9 cells as a model for studying Leydig cell steroidogenesis at the molecular level, we first investigated the effect of hCG on mRNA levels of the steroidogenic enzymes. P-450 scc, 3 beta HSD and P-450 17 alpha were expressed constitutively. The addition of 10 ng/ml hCG enhanced mRNA levels for the three genes within 2 h. Maximal accumulation of P-450 scc, P-450 17 alpha and 3 beta HSD mRNA in treated cells represents a 2.5-, 8.5- and 4-fold increase over control values, respectively. P-450 17 alpha expression reached a maximum by 4 h and then declined rapidly to return to control value by 24 h. The pattern of LH/CG receptor mRNAs in K9 cells was very similar to that of MA10 Leydig cells and showed six transcripts of 1.1, 1.6, 1.9, 2.6, 4.2 and 7.0 kb. Treatment of cells with R or RA resulted in a time- and dose-dependent decrease in all six species.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Inhibition of luteinizing hormone-human chorionic gonadotropin binding by retinoids in a Leydig cell line.

Treatment of K9 mouse Leydig cells with 3 x 10(-6) M retinol (R) and retinoic acid (RA) resulted in 75% and 65% reduction of 125I-labeled hCG binding respectively, when assayed at 35 degrees C. This effect was dose-dependent and was first detected 12 h after initiation of treatment: it was maximal at 48 h for RA. R and RA had no significant effect on the rate of internalization and degradation of 125I-hCG as measured by disappearance of acid-releasable (i.e. surface-bound) radioactivity from the cells and by the appearance of trichloracetic acid-soluble label in the medium. When exposed to increasing concentrations of hCG for 24 h, both retinoid-treated and control cells 'down-regulated' their gonadotropin receptors with the same dose-dependent pattern. The kinetics of reappearance of the receptors was similar for retinoid-treated and control cells, but for treated cells the maximal number of receptors reinitiated at 24 h never exceeded 40% of the values observed with control cells. Scatchard plot analysis confirmed a decrease in hCG receptor number from approximately 26,000 to approximately 6400 and approximately 3500 sites per cell after R and RA treatment. Kd values for 125I-hCG binding were 2 x 10(-10) M, 7.3 x 10(-11) M and 6.9 x 10(-11) M for control, R- and RA-treated cells respectively. On the basis of our data it is likely that retinoid-induced reduction in 125I-hCG binding to K9 Leydig cells is due to decreased receptor synthesis.

Animals↗