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Biomedical subjects

F Denis

Publications and source records attributed to F Denis.

At least 73 records · Page 4Linked to original sources
PubMed1998

[Vaccination against hepatitis B in France in 1996].

A national campain aimed at promoting immunization against hepatitis B was launched in 1994. Two years later, a survey was designed to estimate the situation of the hepatitis B vaccination in France. Around ninety per cent of physicians were hepatitis B vaccinated. The rate of vaccinated population among public hospitals personnel were more than 90% and lower in private hospitals (79%). A quarter of the general population was vaccinated and the higher rate (69%) was observed in the 13-20 years young people. Progressively the contribution of the physicians to the vaccination was increasing (89% in 1996). After a selective vaccination of risk groups and the instauration of the prevention of mother/infant transmission, the French programme was extended to cover adolescents and children. The success of the hepatitis B vaccination campain was obtained by the promotion message toward population and physicians.

Adolescent↗
PubMed1997

Specific activation of the cysteine protease CPP32 during the negative selection of T cells in the thymus.

Cysteine proteases of the CED-3 and ICE family have been recently proposed as the ultimate executioners in several mammalian cell death pathways. Among them, the cysteine protease CPP32 has been shown to participate in programmed cell death (PCD), or apoptosis, affecting lymphoid cells in vitro. In the thymus, negative selection is a mechanism through which developing thymocytes expressing a TcR with high affinity for self peptide-MHC complexes are eliminated by PCD. In order to investigate the role of CPP32 in thymic apoptosis, isolated thymocytes were submitted to cell surface CD3 crosslinking by immobilized anti-CD3 mAb or to dexamethasone treatment. Although apoptosis occurred in the absence or after crosslinking with anti-CD3 mAb, specific activation of CPP32, as assessed by the extent of proteolytic cleavage of the p32 zymogen, was only detected in thymocytes cultured in the presence of the immobilized antibody or dexamethasone. This activation was a very early event during apoptosis as it occurred before the exposure of phosphatidyl serine to the upper side of the cell membrane. This was observed both in anti-CD3- and dexamethasone-induced apoptosis. Moreover, using mice transgenic for pigeon cytochrome C (PCC)-specific TcR, we were able to show that, after injection of PCC, the activation of CPP32 and cleavage of its substrate occurred in thymocytes obtained from mice expressing a permissive MHC haplotype for PCC presentation (H-2k). Moreover, PCC induced apoptosis was blocked by the caspase inhibitor zVAD. While spontaneous apoptosis was not accompanied by detectable levels of CPP32 processing, it was characterized by the proteolysis of poly(ADP-ribose) polymerase (PARP) and was blocked by the cysteine protease inhibitor, zVAD-CH2F. Taken together, these results support the concept that CPP32 is among the earliest effectors of the pathway leading to negative selection of autoreactive thymocytes. Our results also suggest the involvement of a distinct CPP32-like cysteine protease in spontaneous apoptosis of thymocytes.

Animals↗
PubMed1997

The large subunit of replication factor C is a substrate for caspase-3 in vitro and is cleaved by a caspase-3-like protease during Fas-mediated apoptosis.

Caspase-3 is an ICE-like protease activated during apoptosis induced by different stimuli. Poly(ADP-ribose) polymerase (PARP), the first characterized substrate of caspase-3, shares a region of homology with the large subunit of Replication Factor C (RF-C), a five-subunit complex that is part of the processive eukaryotic DNA polymerase holoenzymes. Caspase-3 cleaves PARP at a DEVD-G motif present in the 140 kDa subunit of RF-C (RFC140) and evolutionarily conserved. We show that cleavage of RFC140 during Fas-mediated apoptosis in Jurkat cells and lymphocytes results in generation of multiple fragments. Cleavage is inhibited by the caspase-3-like protease inhibitor Ac-DEVD-CHO but not the caspase-1/ICE-type protease inhibitor Ac-YVAD-CHO. In addition, recombinant caspase-3 cleaves RFC140 in vitro at least at three different sites in the C-terminal half of the protein. Using amino-terminal microsequencing of radioactive fragments, we identified three sites: DEVD723G, DLVD922S and IETD1117A. We did not detect cleavage of small subunits of RF-C of 36, 37, 38 and 40 kDa by recombinant caspase-3 or by apoptotic Jurkat cell lysates. Cleavage of RFC140 during apoptosis inactivates its function in DNA replication and generates truncated forms that further inhibit DNA replication. These results identify RFC140 as a critical target for caspase-3-like proteases and suggest that caspases could mediate cell cycle arrest.

Apoptosis↗
PubMed1997

Adult spinal deformity and respiratory failure. Surgical results in 35 patients.

STUDY DESIGN: Retrospective chart and complementary study review. OBJECTIVES: To describe the features of adult patients with spinal deformity and respiratory failure and to analyze the results of surgical treatment. SUMMARY OF BACKGROUND DATA: Many authors have studied the relation between spinal deformities and cardiorespiratory failure, but there exists little information about the benefits of reconstructive surgery in severely compromised patients. METHODS: The charts and complementary studies of 35 adult patients surgically treated between January 1, 1978, and December 31, 1994, were reviewed. The patients were 18 years old or older (average age, 36 years). They had spinal deformity of any etiology with respiratory insufficiency as evidenced by vital capacity of less than 60% of predictive normal, PaO2 less than 80 mm Hg, or PaCO2 more than 45 mm Hg. All had reconstructive spinal surgery in an attempt to improve their respiratory problem. RESULTS: Seven patients died within the first postoperative year, and one patient was lost to follow-up at 6 months. The other 27 patients had a mean follow-up time of 72 months. The 34 patients were divided into three groups: good, fair, and poor evolution. The patients in the good evolution group had a better preoperative general condition, had more correction of their deformities, had more improvement in their respiratory function, and had fewer complications than those in the other groups. The patients in the poor evolution group were older, had more cardiac problems, and had less correction at surgery. CONCLUSION: The results of surgery varied from extremely good to extremely bad. The seven patients who died within the first year had no benefit, but the 27 others did very well, usually gaining significant improvement of their respiratory function. Because the alternative to surgical correction is death, this study shows that, under the right circumstances, correction of spinal deformity and, therefore, correction of respiratory function can be life-saving.

Adult↗
PubMed1997

Evidence for rapid disappearance of initially expanded HIV-specific CD8+ T cell clones during primary HIV infection.

Down-regulation of the initial burst of viremia during primary HIV infection is thought to be mediated predominantly by HIV-specific cytotoxic T lymphocytes, and the appearance of this response is associated with major perturbations of the T cell receptor repertoire. Changes in the T cell receptor repertoire of virus-specific cytotoxic T lymphocytes were analyzed in patients with primary infection to understand the failure of the cellular immune response to control viral spread and replication. This analysis demonstrated that a significant number of HIV-specific T cell clones involved in the primary immune response rapidly disappeared. The disappearance was not the result of mutations in the virus epitopes recognized by these clones. Evidence is provided that phenomena such as high-dose tolerance or clonal exhaustion might be involved in the disappearance of these monoclonally expanded HIV-specific cytotoxic T cell clones. These findings should provide insights into how HIV, and possibly other viruses, elude the host immune response during primary infection.

Amino Acid Sequence↗
PubMed1997

Paracrine transfer of mouse mammary tumor virus superantigen.

Transfer of vSAG7, the endogenous superantigen encoded in the Mtv7 locus, from MHC class II to MHC class II+ cells has been suggested to occur both in vivo and in vitro. This transfer usually leads to the activation and deletion of T cells expressing responsive V beta s. However, there is no direct molecular evidence for such a transfer. We have developed an in vitro system which confirms this property of vSAGs. vSAG7 was transfected into a class II murine fibroblastic line. Coculture of these cells with class II+ cells and murine T cell hybridomas expressing the specific V beta s led to high levels of IL-2 production which was specifically inhibited by vSAG7- and MHC class II-specific mAbs. Moreover, injection of vSAG7+ class II+ cells in mice led to expansion of V beta 6+ CD4+ cells. We show that this transfer activity is paracrine but does not require cell-to-cell contact. Indeed, vSAG7 was transferred across semi-permeable membranes. Transfer can occur both from class II+ and class II+ cells, indicating that MHC class II does not sequester vSAG7. Finally, competition experiments using bacterial toxins with well defined binding sites showed that the transferred vSAG7 fragment binds to the alpha 1 domain of HLA-DR.

Animals↗
PubMed1997

Accumulation of human immunodeficiency virus-specific cytotoxic T lymphocytes away from the predominant site of virus replication during primary infection.

Down-regulation of the initial burst of viremia during primary human immunodeficiency virus (HIV) infection is thought to be mediated predominantly by HIV-specific CD8+ cytotoxic T lymphocytes (CTL). This response is associated with major perturbations in the T cell receptor (TCR) repertoire. To investigate the failure of the cellular immune response to adequately control viral spread and replication and to prevent establishment of HIV infection, changes in the TCR repertoire and in the distribution of virus-specific CTL between blood and lymph node were analyzed in three patients with primary infection. By the combined use of clonotype-specific polymerase chain reaction and analysis of the frequency of in vivo activated HIV-specific CTL, it was shown that HIV-specific CTL clones preferentially accumulated in blood as opposed to lymph node. Accumulation of HIV-specific CTL in blood occurred prior to effective down-regulation of virus replication in both blood and lymph node. These findings should provide new insights into how HIV, and possibly other viruses, elude the immune response of the host during primary infection.

Antigen Presentation↗
PubMed1997

An anatomical study of the motor distribution of the mandibular nerve for a masseteric-facial anastomosis to restore facial function.

On account of the complex anatomy at the base of skull, surgery here may result in post operative cranial n. deficits. Facial palsy is often feared and its effects upon the patient's psychological and emotional well-being can be catastrophic. The modest results and the side effects of the facio-hypoglossal anastomosis used for facial rehabilitation have led us to consider an anastomosis between a motor branch of the trigeminal n. and the facial n. Dissection has allowed us to demonstrate that the masseteric n. offers the characteristics and the relationships which should make such an anastomosis feasible.

Cadaver↗
PubMed1997

[Emergence of Enterobacter aerogenes strains producing derepressed cephalosporinase at CHU in Limoges: molecular epidemiology by the RAPD technique].

From the beginning of the year 1996, an increase of multiresistant Enterobacter aerogenes isolates was observed. These strains overproduced their chromosomally cephalosporinase. From February to September 1996, we studied the genotypic diversity of multiresistant E. aerogenes isolates. Fifty-seven strains were analysed with the RAPD technique. These strains were isolated from 38 patients in 25 different units. These isolates were collected from urines (37), respiratory tracts (14) and wounds (6). We have observed two distinct profiles. The predominant profile (A) was recovered from 55 strains (36 patients) and the second profile (B) from 2 strains (2 patients). The clonal relatedness of strains isolated (profile A) showed an epidemic spread in our hospital. The RAPD technique is easy to perform and gives results in less than 48 h with a good discriminatory power.

Cephalosporinase↗
PubMed1997

[Primary cytomegalovirus infection and pregnant women: epidemiological study on 1100 women at Limoges].

During 13 months, 1101 women attending for antenatal care at Limoges University Hospital were prospectively studied to determine the prevalence of Cytomegalovirus (CMV) antibodies: 47.9% of these women were CMV seropositive. Ethnic group was strongly associated with CMV status: 42.6% of metropolitan and 94.5% of immigrant women were seropositive. Seropositivity was associated with increasing parity and older age. The risk of a susceptible woman to acquire CMV infection during pregnancy is 0.7%. Among the 4 seroconversions, 3 children were infected, with developmentally and neurologically normal status in one case, moderate ear damage in an other case and a disseminated infection in the third case requiring a provoked abortion.

Adult↗
PubMed1997

[Pulmonary infection caused by Fusobacterium nucleatum].

Thoracic infection by anaerobes are uncommon diseases and often presents difficulty in diagnosis. We report a case of thoracic infection due to Fusobacterium Nucleatum. A 60-year old man presented a lesion infiltrating from the lung to the thoracic wall. Fusobacterium Nucleatum was isolated on pus collected. Treatment by penicillin, metronidazole combined with surgical drainage was highly effective.

Anti-Bacterial Agents↗
PubMed1997

[Seroprevalence of HBV, HCV and HDV hepatitis markers in 500 patients infected with the human immunodeficiency virus].

The serological status for both hepatitis B, C and D viruses was analyzed for 500 HIV seropositive patients and for 1037 of a control group. The prevalence was 31.4% for anti HCV, 13.8% for HBs Ag and 69.0% for one or more HBV markers in HIV positive patients and respectively 2.5%, 2.7% and 13.1% in control group. The markers for hepatitis D were founded among 21% of the HBs Ag carriers (patients and control group), correlated with drug i.v. use. The prevalence of anti-HCV was 71.6% in subjects who had blood-borne HIV infection and 1.5% in those with sexually acquired infection. The prevalence in control group was 10.2% and 1.7% respectively according to the same risk factors. The prevalence of HBs Ag was higher among HIV positive patients with sexual risk (17.5%) than with blood exposition (9.9%) and a variation in the same direction is observed in control group (3% v.s. 1%). The relation between markers for hepatitis B and hepatitis C was negative.

Adolescent↗
PubMed1996

Diversity of antibody binding to V3 peptides representing consensus sequences of HIV type 1 genotypes A to E: an approach for HIV type 1 serological subtyping.

We investigated whether V3-binding assays might be useful to analyze human immunodeficiency virus type 1 (HIV-1) variants in different geographic regions. We showed that strong cross-reactivity between subtype-specific V3 peptides is almost inevitable in standard indirect enzyme-linked immunosorbent assays (EIA), impairing precise serological subtyping. We therefore developed a subtype-specific EIA (HIV-1 SSEIA) that uses the principle of blocking by an excess of peptide in the liquid phase. Using 231 serum samples collected from HIV-1-infected individuals in 10 different geographical areas from 4 continents, we showed that this approach detected the dominant subtype reactivity in more than 97% of the cases. Internal controls (0 and 100% blocking) were used for every sample such that comparative analysis was possible, independent of both the individual humoral response and the time of collection during the course of infection. This was validated by the excellent concordance of the serological profiles of couples and the temporal stability of the serological profile in individuals. The geographical distribution of the various subtypes in the SSEIA was in agreement with the present knowledge of the distribution of the various genotypes. Although the goal of this study was not an extensive seroepidemiological survey, our results showed that the various profiles in most of the regions were relatively homogeneous, but in central Africa there was a large diversity of serological profiles. Cluster analysis identified a limited number of V3 serogroups of serotypes within the HIV-1 group M. Five serogroups, some of them divided into subgroups, were identified and characterized by a mean serological profile. Our data confirmed that subtypes A and C, although being dissimilar genetic subtypes, present conserved antigenic properties in the V3 region, and that the D subtype is probably the most divergent within the group M (B Korber et al., J Virol 1994;68:6730). Cluster analysis showed a clear correlation between position within the dendrogram and geographical origin of the samples. This is further support for the reliability and thereof the usefulness of the SSEIA. This simple methodology may help facilitate the analysis of the distribution of various HIV-1 subtypes circulating in different populations and regions.

Amino Acid Sequence↗
PubMed1996

Paraplegia resulting from vessel ligation.

STUDY DESIGN: This is a retrospective review of clinical records for evidence of paraplegia specifically resulting from segmental vessel ligation during anterior spinal surgery. OBJECTIVES: To determine the precise risk rate, and to potentially identify risk factors. SUMMARY OF BACKGROUND DATA: Although many authors have alluded to this risk, the exact risk rate and risk factors have never been identified. METHODS: All patients having an anterior approach involving T1-L3 were reviewed. The two reviewers were not involved in any of the surgeries. The 1197 cases were consecutive from 1967 to 1991. RESULTS: There were no paralyses. CONCLUSIONS: There would appear to be virtually no risk to segmental vessel ligation provided: 1) vessel ligation is unilateral, 2) done on the convexity of a scoliosis, 3) ligated at midvertebral body level, and 4) hypotensive anesthesia is avoided. Soft clamping with somatosensory-evoked potential monitoring does not appear justified.

Adult↗
PubMed1996

New vectors for combinatorial deletions in yeast chromosomes and for gap-repair cloning using 'split-marker' recombination.

New tools are needed for speedy and systematic study of the numerous genes revealed by the sequence of the yeast genome. We have developed a novel transformation strategy, based on 'split-marker' recombination, which allows generation of chromosomal deletions and direct gene cloning. For this purpose, pairs of yeast vectors have been constructed which offer a number of advantages for large-scale applications such as one-step cloning of target sequence homologs and combinatorial use. Gene deletions or gap-repair clonings are obtained by cotransformation of yeast by a pair of recombinant plasmids. Gap-repair vectors are based on the URA3 marker. Deletion vectors include the URA3, LYS2 and kanMX selection markers flanked by I-Scel sites, which allow their subsequent elimination from the transformant without the need for counter-selection. The application of the "split-marker' vectors to the analysis of a few open reading frames of chromosome XI is described.

Chromosomes, Fungal↗
PubMed1996

Evaluation of rapid culture centrifugation method for adenovirus detection in stools.

Routine laboratory testing for adenovirus (Adv) requires a procedure that is rapid and reliable, especially for samples from children and immunosuppressed patients, when diarrhea may signal the onset of severe gastrointestinal disorders. An improved culture technique for Adv isolation, using centrifugation step of 24-well plates and needing only 48 h incubation, was evaluated for 382 stool samples. This technique was compared with conventional tube cell culture and a commercial enzyme-linked immunosorbent assay (ELISA) kit. Adv was isolated in 36 samples (9.4%) by rapid culture, in 32 (8.4%) by conventional culture, and in 42 samples (11%) using genus-specific ELISA. A total of 30 isolates were found to be Adv positive in both rapid and conventional cultures, and half of the Adv-positive rapid culture isolates were identified as serotypes 40/41 using a type-specific ELISA. The improved culture method considerably reduces incubation time and also offers a slightly enhanced sensitivity to Adv serotypes. Combined with appropriate cell lines adapted to the isolation of enteric adenoviruses, it therefore constitutes a valuable laboratory test particularly useful in the diagnosis of gastroenteritis.

Adenovirus Infections, Human↗