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Biomedical subjects

R Lévy

Publications and source records attributed to R Lévy.

At least 19 recordsLinked to original sources
PubMed2006

[How to improve assisted reproductive techniques results? Biological aspects].

In spite of recent improvements in IVF, pregnancy rates remained dramatically low. Better criteria are therefore necessary to establish the viability of a transferable top quality embryo. Observation of both pronuclei patterns at the zygote stage and early cleavage appears to offer an additional prognostic tool, correlating with chromosomal status and IVF outcome. Developmental ability and morphological evaluation of the embryo on days 2-3 remains the most used and valid method of selection, even though it is not sufficient to select embryos with the higher implantation potential. Blastocyst culture is another possible strategy for selecting the ideal embryo with theoretical, reduced risk of aneuploidies, though not all major chromosomal aberrations are excluded by prolonged in-vitro-culture. Concerning micro-manipulation of embryo such as hatching or cleaning or cytoplasm transfer, only controversial data are still available. In the field of ART, recent meta-analysis could not conclude to a positive effect concerning hatching, blastocyst culture or systematic PGD-AS (preimplantation genetic diagnosis with aneuploidy screening). Beside the technological aspect, Quality Management System is of great importance. Staff development and monitoring of staff performance in the IVF laboratory has to be considered in the next future.

Blastocyst↗
PubMed2006

Seminal plasma promotes the attraction of Langerhans cells via the secretion of CCL20 by vaginal epithelial cells: involvement in the sexual transmission of HIV.

BACKGROUND: Heterosexual human immunodeficiency virus (HIV) transmission implies the crossing of the vaginal mucosa by virions present in the semen, potentially using Langerhans cells as transporters. The recruitment of these cells in the mucosa is mediated by the chemokine macrophage inflammatory protein 3alpha (CCL20). The aim of this study was to evaluate the capacity of the semen to induce Langerhans cell recruitment via the production of CCL20 by vaginal epithelial cells. METHODS AND RESULTS: Using a vaginal epithelium model based on the SiHa cell line and human seminal plasma, we demonstrated that semen enhanced the production of CCL20. This secretion was regulated by the nuclear factor-kappaB intracellular signalling pathway. Fractionation of the seminal plasma indicated that the secretion of CCL20 was stimulated by high molecular weight compounds present in semen. Migration assays demonstrated that secreted CCL20 was able to promote the recruitment of Langerhans cell precursors (LCps), which remain permissive to X4 and R5 HIV infection. CONCLUSIONS: Our results demonstrate that epithelial cells respond to factors present in semen by secreting CCL20, leading to the enhancement of LCp recruitment. These data argue in favour of the implication of epithelial cells in the heterosexual transmission of HIV.

Cell Movement↗
PubMed2006

Protein-GAG interactions: new surface-based techniques, spectroscopies and nanotechnology probes.

New approaches, rooted in the physical sciences, have been developed to gain a more fundamental understanding of protein-GAG (glycosaminoglycan) interactions. DPI (dual polarization interferometry) is an optical technique, which measures real-time changes in the mass of molecules bound at a surface and the geometry of the bound molecules. QCM-D (quartz crystal microbalance-dissipation), an acoustic technique, measures the mass and the viscoelastic properties of adsorbates. The FTIR (Fourier-transform IR) amide bands I, II and III, resulting from the peptide bond, provide insight into protein secondary structure. Synchrotron radiation CD goes to much shorter wavelengths than laboratory CD, allowing access to chromophores that provide insights into the conformation of the GAG chain and of beta-strand structures of proteins. To tackle the diversity of GAG structure, we are developing noble metal nanoparticle probes, which can be detected at the level of single particles and so enable single molecule biochemistry and analytical chemistry. These new approaches are enabling new insights into structure-function relationships in GAGs and together they will resolve many of the outstanding problems in this field.

Animals↗
PubMed2005

Specific molecular interactions by force spectroscopy: from single bonds to collective properties.

Interactions involving multiple bonds occur throughout biology, and have distinct properties that are fundamentally different from those present in single bond systems. We have developed a new method to analyse the AFM force measurements in order to extract relevant information and to characterise the interactions involving from single to multiple bonds. Our study reveals a surprising behaviour in the presence of multiple bonds with a high rebinding probability: the mean binding forces increase with decreasing pulling velocity. Such behaviour is different from the force dependence on the loading rate for single bond rupture or existing models for multiple bonds rupture.

Biophysical Phenomena↗
PubMed2005

[Apoptosis in oocyte].

Apoptosis has been reported in oocyte and preimplantation embryo. The developmental potential of embryo could be related to its rate of apoptosis. This apoptotic fate could be modulated by suboptimal culture conditions: however, a critical role is played by gamete quality and in particular, oocyte apoptosis. Thus, the investigation of apoptosis-related genes and mechanisms in oocyte is crucial. New technologies, such as microarrays, may contribute to the elucidation of molecular pathways involved in oocyte survival and maturation. Besides in vitro culture and ART, the study of oocyte apoptosis could benefit to research in the fields of menopause or cancer treatment.

Animals↗
PubMed2005

[Impact of sperm DNA fragmentation on ART outcome].

We have used the sperm chromatin structure assay (SCSA) test in order to determine if correlations can be found between sperm DNA fragmentation and spermogram parameters. Only necrospermia and DNA fragmentation index are strongly correlated (P<0.0001). Neither fertilization rates for ICSI and IVF, nor blastocyst formation rates are impaired by a high DFI. However when the critical DFI>30% is reached, the chances of having ongoing pregnancies after blastocyst transfer are reduced by three. Treatments with antioxidants are of limited efficacy even though we obtained 2 deliveries after DFI treatments with such treatments. New strategies in order to improve the pregnancy rates for these peculiar cases of reduced fertility are discussed.

Adult↗
PubMed2005

New tools for force spectroscopy.

Force curves using atomic force microscopy have been proposed as a new tool to probe the conformation of adsorbed polymers and interactions between molecules through the analysis of the rupture distributions. We describe an algorithm for the computer-assisted detection of ruptures in force curves. This program allows us to automatically detect the ruptures. The algorithm is based on an analysis of the standard deviation in a sliding window along the length of the force. Automatic detection of ruptures constitutes an important step forward because the time required to analyse the curves is significantly reduced, thus allowing the user to perform multiple experiments. In addition, using these tools we can address the problem of the interpretation of rupture distributions.

Adsorption↗
PubMed2004

Effects of cryopreservation on the meiotic spindle of human oocytes.

The microtubular meiotic spindle of most mammals, including humans, is very sensitive to cooling [Hum. Reprod. 16 (2001) 2374; Fertil. Steril. 54 (1990) 102; Fertil. Steril. 75 (2001) 769; Zygote 3 (1995) 357] and is rapidly depolymerised even after a slight reduction in temperature to 33 degrees C. Spindle disassembly is dependent on the extent of temperature decrease and its duration. After rewarming, the recovery is far from complete. Cryoprotectants themselves may alter the spindle structure, depending on the duration and temperature of exposure, the duration of recovery at 37 degrees C and the species [Hum. Reprod. Update 2 (1996) 193]. Damage to the meiotic spindle is considered to be the cause of aneuploid embryos, by inducing chromatid non-disjunction and chromosome scattering and by disturbing the sequence of events leading to the completion of meiosis and fertilisation. Nevertheless, a consensus arose from all the studies: appropriate exposure to cryoprotectants and appropriate rates of cooling and thawing allow the cryopreservation of mature oocytes without any significant changes in their second meiotic spindle organisation and without any increase in the rate of aneuploid embryos [Mol. Hum. Reprod. 2 (1996) 445; Hum. Reprod. 8 (1993) 1101; Hum. Reprod. 9 (1994) 684; Microsc. Res. Technol. 27 (1994) 165; Fertil. Steril. 75 (2001) 354]. These fundamental studies in humans, showing good preservation of cell structures after freeze-thaw procedures opened the way to new successful clinical trials with embryos derived from cryopreserved mature oocytes [Fertil. Steril. 68 (1997) 724]. Considering immature oocyte freezing at prophase I (germinal vesicle (GV) stage), a stage which was thought to be less sensitive to cryoinjury, pooled data from the literature showed no advantage in terms of survival rates, fertilisation rates of in vitro matured oocytes and developmental ability of the resulting embryos, especially in unstimulated cycles. Moreover, conflicting results are reported on the effects of freezing on the spindle-chromosome configuration of immature oocytes or in vitro matured oocytes, highlighting the need for large scale studies [Hum. Reprod. 10 (1995) 1816; Hum. Reprod. 13 (Suppl. 3) (1998) 161; Hum. Reprod. 17 (2002) 1885; Microsc. Res. Technol. 27 (1994) 165; Fertil. Steril. 68 (1997) 920]. One child has been born after the use of cryopreserved immature oocytes at GV stage, matured in vitro and fertilised by ICSI [Hum. Reprod. 13 (1998) 3156], demonstrating at least the feasibility of this technique. Improvements are required so as to make mature and immature oocyte cryopreservation an established and safe technique for ART.

Cold Temperature↗
PubMed2002

Clinical and genetic studies of Birt-Hogg-Dubé syndrome.

Birt-Hogg-Dubé syndrome (BHD) is an autosomal dominant cancer syndrome characterised by benign skin tumours, renal tumours, and spontaneous pneumothorax. The gene has been mapped to chromosome 17p11.2 and recently identified, expressing a novel protein called folliculin. We report the clinical and genetic studies of four sporadic BHD cases and four families with a total of 23 affected subjects. Haplotype analysis of these families using BHD linked markers showed they did not share the same affected alleles, excluding common ancestry. Mutation analysis of the BHD gene identified two germline mutations on exon 11 (c.1733insC and c.1733delC) in three of four families as well as two of four sporadic cases. A novel somatic mutation, c.1732delTCinsAC, was detected in a BHD related chromophobe renal carcinoma. Our results confirmed the (C)8 tract in exon 11 as a mutational hot spot in BHD and should always be considered for future genetic testing. Our observation also indicated that the second hit (of Knudson's two hit theory) in some BHD related tumours is in the form of somatic mutation rather than LOH. In a large French family in which eight affected subjects carry the c.1733delC mutation, a phenocopy who has multiple episodes of spontaneous pneumothorax was identified. A total of five mutation carriers (aged between 37 to 66) did not have any evidence of BHD features, suggesting either reduced penetrance or late age of onset of the disease. In addition, six out of eight affected subjects who have positive germline mutation have confirmed neoplastic colonic polyps, indicating that colorectal neoplasia is an associated feature of BHD in some families. Our studies have observed several interesting genetic features in BHD: (1) the poly (C) tract in exon 11 as a mutational hot spot; (2) the existence of phenocopy; (3) reduced penetrance or late age of onset of disease; (4) association with colorectal neoplasia in some families; and (5) somatic mutation instead of LOH as the second hit in BHD tumours.

Adult↗
PubMed2002

Preclinical development of a vaccine 'against smoking'.

BACKGROUND: Nicotine is the main culprit for dependence on tobacco-containing products, which in turn are a major etiologic factor for cardiovascular diseases and cancer. This publication describes a vaccine, which elicits antibodies against nicotine. The antibodies in the blood stream intercept the nicotine molecule on its way to its receptors and greatly diminish the nicotine influx to the brain shortly after smoking. METHODS: The nicotine molecule is chemically linked to cholera toxin B as a carrier protein in order to induce antibodies. The potential to elicit antibodies after subcutaneous as well as intranasal immunization is evaluated. In order to simulate realistic conditions, nicotine pumps delivering the nicotine equivalent of 5 packages of cigarettes for 4 weeks are implanted into the mice 1 week prior to vaccination. The protective effect of the vaccine is measured 5 weeks after vaccination by comparing the influx of radiolabeled nicotine in the brains of vaccinated and non-vaccinated animals 5 min after challenge with the nicotine equivalent of 2 cigarettes. RESULTS: The polyclonal antibodies induced by the vaccine show a mean avidity of 1.8 x 10(7) l/Mol. Subcutaneous immunization elicits high antibody levels of the IgG class, and significant IgA antibody levels in the saliva of vaccinated mice can be found after intranasal vaccination. The protective effect also in the animals with implanted nicotine pumps is significant: less than 10% of radiolabeled nicotine found in the brains of non-vaccinated animals can be found in the brains of vaccinated animals. CONCLUSIONS: These data provide credible evidence that a vaccine can break the vicious circle between smoking and instant gratification by intercepting the nicotine molecule. Astonishingly, there is no sign of exhaustion of specific antibodies even under extreme conditions, which makes it highly unlikely that a smoker can overcome the protective effect of the vaccine by smoking more. Finally, the high titers of specific antibodies after 1 year let us hope that booster vaccinations are probably only necessary in intervals of years.

Administration, Intranasal↗
PubMed1996

Evaluation and comparison of PCR and hybridization methods for rapid detection of cytomegalovirus in clinical samples.

Rapid diagnosis of cytomegalovirus (CMV) infection may be obtained by molecular techniques, such as the polymerase chain reaction (PCR) and hybridization assays. The optimal technique to detect CMV in clinical samples was assessed. Two different PCR assays were used, targeting either the major immediate early 1 (MIE 1) or the HXLF 4 gene. The PCR products were detected by gel electrophoresis, dot blotting and an easy to use, rapid, solid phase hybridization assay, DNA enzyme immunoassay (DEIA). Standard tissue culture was also used. Cerebrospinal fluids (18), liver biopsies (9) from hepatic transplant recipients, amniotic fluids (7) from mothers with suspected peripartum infection, and samples (6) of miscellaneous origin (brain and fundus biopsy, pericardial and pleural fluid) were tested. Among the 40 samples, CMV was detected in 19 cases. Three were positive by both molecular techniques and tissue culture, 14 by molecular methods and 2 by culture. 16/19 or 9/19 CMV-positive samples were detected by PCR amplification of the HXLF 4 or MIE 1 gene, respectively and 14/16 HXLF 4-positive samples were detected using either dot-blot or DEIA, compared to 9/16 using gel electrophoresis. Thus, the most sensitive assays for the detection of CMV in clinical samples using the methods compared in the current study were PCR amplification of the HXLF 4 gene followed by dot-blot or DEIA hybridization.

Amniotic Fluid↗
PubMed1995

Human immunodeficiency virus associated Hodgkin's disease: report of 45 cases from the French Registry of HIV-Associated Tumors.

Clinical and pathological characteristics as well as outcome of 45 Hodgkin's disease (HD) cases collected by the French registry of HIV-associated tumors between January 1987 and December 1989 (all clinically staged according to the Ann Arbor system) were analyzed and compared with those of a cohort of 407 patients with clinical stages (CS) IA to IVB enrolled between September 1981 and August 1988 in a multicentric clinical trial. The route of HIV infection, initial CD4 cell count at the time of HD diagnosis and CDC class of HIV infection were studied as well as the progression to AIDS onset were recorded. HIV-HD is characterized by a predominance of advanced CS (75%), B symptoms (80%) and mixed cellularity histology (49%), as well as by early bone marrow involvement (24%); a specific feature is the rare occurrence of mediastinal involvement (13% in HIV-HD versus 71% in primary HD). With standard therapies, 79% of the patients achieved a complete remission, but hematological and infectious complications were very frequent. The proportion of intravenous drug abusers (IVDA) in HIV-HD (38%) is higher than in French HIV-infected population as a whole (20.8%). Median CD4 cell count was 306/microliters at the time of HD diagnosis, while only 5 cases (11%) were preceded by an AIDS manifestation; progression to AIDS rate was 94% at 2 years. Overall 2-year survival was 41%, with 71% for patients with an initial CD4 cell count over 300/microliters and 0% for those with CD4 cell count lower than 300/microliters (p < 0.01); opportunistic infections were the most frequent cause of death. HIV-HD seems to occur preferentially in.(ABSTRACT TRUNCATED AT 250 WORDS)

AIDS-Related Opportunistic Infections↗
PubMed1977

[Myocardial infarction and heterozygous sickle cell anemia. Apropos of 2 cases].

Thromboses are a classical complication of sickle cell disease in the severe homozygous form. In the heterozygous individual, although the risk of severe thromobotic episodes is small, it must nevertheless be recognised. The authors report two cases of myocardial infarction in patients whose coronary arteries were relatively free from atherosclerosis; they were young men, with the heterozygous form of sickle cell disease AS. The part played by inheritance, the factors favouring vascular occlusion, and the possibility of preventing such episodes are all discussed.

Adult↗