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C Leroux

Publications and source records attributed to C Leroux.

At least 37 records · Page 2Linked to original sources

Lipoprotein lipase activity and mRNA are up-regulated by refeeding in adipose tissue and cardiac muscle of sheep.

Previous studies in rodents have shown that the lipoprotein lipase (LPL) regulation is complex and often opposite in adipose tissue (AT) and muscle in response to the same nutritional treatment. However, neither LPL responses nor the molecular mechanisms involved in the nutritional regulation have been studied in both AT and muscle of ruminant species. To explore this, we measured the LPL activity and mRNA levels in perirenal AT and cardiac muscle (CM) of control, 7-d-underfed or 14-d-refed ewes. Underfeeding decreased (P < 0.01) LPL activity both in AT (-59%) and CM (-31%), and these activities were restored (P < 0.01) by refeeding (AT, +248%; CM, +34%). Variations of LPL mRNA level measured by real-time reverse transcription-polymerase chain reaction or by Northern blot followed variations of LPL activity: underfeeding decreased AT- and CM-LPL mRNA levels (-58 and -53%, respectively), and refeeding restored (P < 0.01) them in CM (+117%) and increased them over the baseline in AT (+640%). Quantification of either 3.4- or 3.8-kb LPL mRNA levels revealed a predominant (P < 0.001) expression of the 3.4-kb mRNA in AT (60%) and of the 3.8-kb mRNA in CM (56%), without any preferential regulation of one of these mRNA species by the nutritional status. This work reveals a tissue-specific expression pattern of the ovine LPL gene and a pretranslational nutritional regulation of its expression, which is achieved in the same direction in perirenal AT and CM. The different regulation of CM-LPL between ewes and rats probably arises from peculiarities of ruminant species for nutrient digestion and absorption and liver lipogenesis.

3' Untranslated Regions↗

Accelerated coronary atherosclerosis and arteriosclerosis in young human-immunodeficiency-virus-positive patients.

OBJECTIVE: To determine the type of lesions observed in young patients infected with human immunodeficiency virus-1 (HIV-1). DESIGN: Examination of coronary networks in corpses of 13 men and two women who had died aged 23-32 years after having been infected with HIV-1 virus, having been seropositive for 2-5 years. Causes of death were infectious complications (five cases), infection with cytomegalovirus leading to gastro-intestinal haemorrhaging (one case), infection with cytomegalovirus and Kaposi's sarcoma (one case), overdoses of drugs (five cases) and sudden death (three cases). METHODS: The pathological analysis was carried out on the proximal and distal coronary networks. In order to characterize the lesions better, the cells and the cytokines involved were characterized by immunohistochemistry. RESULTS: In all 15 cases we observed thickening of intima in the proximal network at least as great as that of the media, caused by a proliferation of secreting cells, phenotypically identified as smooth muscle cells, with exaggerated production of elastic fibres and in association with an increase in the expression of tumor necrosis factor-alpha and interleukin-1 alpha. In nine cases, atherosclerosis had developed from and on the surface of this proliferation and in four cases arteriosclerosis had an unusual appearance, in the form of mamillated vegetations with endoluminal protrusions. A similar proliferation was found in the distal network in four cases, but with a significantly smaller proportion of elastic fibres. CONCLUSIONS: The lesions we examined in these young HIV-1-infected patients presented particular features and were intermediate between the lesions observed during common coronary atherosclerosis and atherosclerosis associated with chronic rejection of cardiac transplants.

Adult↗

The S2 gene of equine infectious anemia virus is a highly conserved determinant of viral replication and virulence properties in experimentally infected ponies.

Equine infectious anemia virus (EIAV) is genetically one of the simplest lentiviruses in that the viral genome encodes only three accessory genes, tat, rev, and S2. Although serological analyses demonstrate the expression of the S2 protein in persistently infected horses, the role of this viral gene remains undefined. We recently reported that the S2 gene is not essential for EIAV replication in primary equine macrophages, as EIAV mutants lacking the S2 gene replicate to levels similar to those of the parental virus (F. Li, B. A. Puffer, and R. C. Montelaro, J. Virol. 72:8344-8348, 1998). We now describe in vivo studies that examine the evolution and role of the S2 gene in ponies experimentally infected with EIAV. The results of these studies reveal for the first time that the S2 gene is highly conserved during persistent infection and that deletion of the S2 gene reduces viral virulence and virus replication levels compared to those of the parental virus containing a functional S2 gene. These data indicate that the EIAV S2 gene is in fact an important determinant of viral replication and pathogenic properties in vivo, despite the evident lack of S2 influence on viral replication levels in vitro. Thus, these observations suggest in vivo functions of EIAV S2 that are not adequately reflected in simple infections of cultured cells, including natural target macrophages.

Amino Acid Sequence↗

Effects of long terminal repeat sequence variation on equine infectious anemia virus replication in vitro and in vivo.

The long terminal repeat (LTR) is reported to be one of the most variable portions of the equine infectious anemia virus (EIAV) genome. To date, however, no information is available on the effects of observed sequence variations on viral replication properties, despite a widespread assumption of the biological importance of EIAV LTR variation. EIAV LTR sequence variability is confined mostly to a small portion of the enhancer within the U3 segment of the LTR. Analysis of published EIAV LTR sequences revealed six different types of LTR based on the pattern of putative transcription factor motifs within the variable region of the enhancer. To test directly the significance of LTR variation, the in vitro and in vivo replication properties of two variant LTR species were investigated using two isogenic viruses, EIAV(19-2) and EIAV(19-2-6A), differing only within the enhancer region. The results of these studies demonstrated that the two variants replicated with similar kinetics and to equal levels in cultured equine fibroblasts or in equine macrophage, the natural target cell of EIAV, even after prolonged serial passage in the latter cell type. Furthermore, EIAV(19-2) and EIAV(19-2-6A) variants demonstrated similar replication levels in experimentally infected ponies. However, ponies infected with EIAV(19-2-6A) exhibited a rapid switch in the prevalent LTR type, such that by 112 days postinfection, no original-LTR-type viruses were evident. This specific and rapid shift in LTR quasispecies indicates an in vivo selection that is not reflected in simple in vitro replication rates, suggesting undefined selection pressures in vivo that drive LTR variation during persistent EIAV infection.

Animals↗

[Postoperative antero-external tibial compartment syndrome: co-responsibility of the operating table].

A 26-year-old, ASA1 patient underwent maxillofacial surgery under general anaesthesia, of 12-hour duration in the supine position. Postoperatively he developed rhabdomyolysis and acute renal failure. In the subsequent days, a bilateral leg compartment syndrome occurred with anterior tibial motor nerve injury requiring fasciotomies and excision of necrotic muscles. Several aetiological factors may have contributed to this accident: a long-lasting procedure, controlled hypotension and inappropriate position of the lower limbs. A laboratory study showed that the hardness of some new operating tables could be responsible for this complication. Some prophylactic measures are therefore required before the use of such devices.

Adult↗

Cross-sectional and 4-year longitudinal associations between brachial pulse pressure and common carotid intima-media thickness in a general population. The EVA study.

BACKGROUND AND PURPOSE: The cross-sectional and 4-year longitudinal associations between brachial pulse pressure (PP) and ultrasound measurements of common carotid intima-media thickness (CCA-IMT) were assessed. METHODS: A population of 957 volunteers aged 59 to 71 years was recruited from the electoral rolls of the city of Nantes (western France) and reexamined 4 years later. Longitudinal changes in PP and CCA-IMT were computed as the difference between 4-year follow-up and baseline values. RESULTS: Baseline CCA-IMT and PP were positively associated in both age- and sex-adjusted analysis (partial correlation coefficient=0.20, P<0.001) and in multivariate analysis adjusted for traditional cardiovascular risk factors and mean blood pressure (partial correlation coefficient=0.18, P<0.001). In longitudinal analysis, baseline PP was associated with the change in 4-year CCA-IMT (partial correlation coefficient=0.11, P<0.001), and baseline CCA-IMT was a predictor of the 4-year change in PP (partial correlation coefficient=0.10, 0.001<P<0.01). No association between mean blood pressure and CCA-IMT was observed once PP was taken into account, in either cross-sectional or longitudinal analyses (partial correlation coefficients ranged from 0.00 to 0.03). Similar patterns of results were observed in hypertensive, nonhypertensive, and antihypertensive-treated and -nontreated subjects. CONCLUSIONS: This longitudinal study of a large population of relatively aged subjects suggests that elevated levels of PP are associated with the progression of CCA-IMT, and increased CCA-IMT is associated with PP widening. The nature of these relationships and whether atherosclerosis progression over time is involved or not in these associations merit further investigations.

Aged↗

Development and characterization of an in vivo pathogenic molecular clone of equine infectious anemia virus.

An infectious nonpathogenic molecular clone (19-2-6A) of equine infectious anemia virus (EIAV) was modified by substitution of a 3.3-kbp fragment amplified by PCR techniques from a pathogenic variant (EIAV(PV)) of the cell culture-adapted strain of EIAV (EIAV(PR)). This substitution consisted of coding sequences for 77 amino acids at the carboxyl terminus of the integrase, the S1 (encoding the second exon of tat), S2, and S3 (encoding the second exon of rev) open reading frames, the complete env gene (including the first exon of rev), and the 3' long terminal repeat (LTR). Modified 19-2-6A molecular clones were designated EIAV(PV3.3), and infection of a single pony (678) with viruses derived from a mixture of five of these molecular clones induced clinical signs of acute equine infectious anemia (EIA) at 23 days postinfection (dpi). As a consequence of this initial study, a single molecular clone, EIAV(PV3.3#3) (redesignated EIAV(UK)), was selected for further study and inoculated into two ponies (613 and 614) and two horses (700 and 764). Pony 614 and the two horses developed febrile responses by 12 dpi, which was accompanied by a 48 to 64% reduction in platelet number, whereas pony 613 did not develop fever (40.6 degrees C) until 76 dpi. EIAV could be isolated from the plasma of these animals by 5 to 7 dpi, and all became seropositive for antibodies to this virus by 21 dpi. Analysis of the complete nucleotide sequence demonstrated that the 3.3-kbp 3' fragment of EIAV(UK) differed from the consensus sequence of EIAV(PV) by just a single amino acid residue in the second exon of the rev gene. Complete homology with the EIAV(PV) consensus sequence was observed in the hypervariable region of the LTR. However, EIAV(UK) was found to contain an unusual 68-bp nucleotide insertion/duplication in a normally conserved region of the LTR sequence. These results demonstrate that substitution of a 3.3-kbp fragment from the EIAV(PV) strain into the infectious nonpathogenic molecular clone 19-2-6A leads to the production of progeny virus particles with the ability to induce clinical signs of EIA. Therefore, EIAV(UK), which is the first pathogenic, cell culture-adapted molecular clone of EIAV to be described, should be of value in identifying viral determinants of pathogenicity.

Amino Acid Sequence↗

Differential splicing of pre-messenger RNA produces multiple forms of mature caprine alpha(s1)-casein.

The identity of multiple forms of caprine alpha(s1)-casein in variants A, B, and C has been determined by structural characterisation using mass spectrometry, automated Edman degradation and peptide mapping. Mature goat alpha(s1)-casein exists as a mixture of at least four molecular species which differ in peptide chain length. The main component corresponds to the 199-residues form already described. The other three, in lesser amounts, were shorter forms of alpha(s1)-casein and differed for the deleted peptides 141-148, as shown previously for ovine alpha(s1)-casein, peptide 110-117, or Gln78. Analysis of alpha(s1)-casein mRNA from milk somatic cells demonstrated that these forms originated from skipping events at the level of exon 13 (codifying for peptide 110-117) and 16 (codifying for peptide 141-148) and from the presence of a cryptic splice site within exon 11 (whose first CAG triplet encodes Gln78) during primary transcript processing. The finding of these splicing abnormalities in the three common variants A, B, and C suggests that this is a general feature of alpha(s1)-casein in goat. A further source of heterogeneity of caprine alpha(s1)-casein was identified in the discrete phosphorylation of seryl residues. Eight serine residues (at positions 44, 46, 64 to 68 and 75) are fully phosphorylated (except in variant A because of the replacement Glu77-->Gln which prevents phosphorylation of Ser75). Conversely, Ser115 and Ser41 are phosphorylated only to about 50% and 20%, respectively. Ser12, although located in a consensus triplet, is never phosphorylated, similarly to the ovine alpha(s1)-casein variants. These results confirm that there are stabilised mechanisms of simultaneous synthesis of alpha(s1)-casein at different length and of post-translational modification in both caprine and ovine species.

Alternative Splicing↗

Genomic heterogeneity of small ruminant lentiviruses: existence of heterogeneous populations in sheep and of the same lentiviral genotypes in sheep and goats.

We have recently shown that French small ruminant lentiviruses (SRLV) from sheep are more similar to Caprine Arthritis Encephalitis Virus (CAEV) than to visna maedi virus (VMV) in a conserved region of the pol gene. To extend these results, we have examined sequences from a variable region of the env gene in French SRLV. We found that they were nearly equally distant from both CAEV and VMV strains, suggesting a considerable divergence since the initial introduction of the virus. Analysis of separate clones from individual animals showed that some carry a population of variant viruses. The study of further pol gene sequences from both goats and sheep suggests that viral variants show little or no host species specificity. A phylogenetic tree of pol gene sequences confirmed the presence of a novel genotype of SRLV in France.

Amino Acid Sequence↗

Presence of a unique parainfluenza virus 3 strain identified by RT-PCR in visna-maedi virus infected sheep.

The presence in farm sheep of a paramyxovirus closely related to parainfluenza virus type 3 (Pi3) from humans or cattle was confirmed using RT-PCR on RNA samples from lung cells from slaughtered animals. Sequencing and restriction enzyme patterns of the amplified fragment of the F gene confirms the distinctness of the isolate, and suitable PCR primers allow specific detection of the ovine virus. A study of the incidence of ovine Pi3 in samples from sheep with or without distinctive histopathological signs of maedi shows that it is uncommon in aged sheep with overt lentiviral disease, but it occurs at moderate frequency in lambs and may, in the presence of visna-maedi virus, contribute to early lesion formation.

Abattoirs↗

Novel and dynamic evolution of equine infectious anemia virus genomic quasispecies associated with sequential disease cycles in an experimentally infected pony.

We have investigated the genetic evolution of three functionally distinct regions of the equine infectious anemia virus (EIAV) genome (env, rev, and long terminal repeat) during recurring febrile episodes in a pony experimentally infected with a well-characterized reference biological clone designated EIAV(PV). Viral populations present in the plasma of an EIAV(PV)-infected pony during sequential febrile episodes (18, 34, 80, 106, and 337 days postinfection) were amplified from viral RNA, analyzed, and compared to the inoculated strain. The comparison of the viral quasispecies showed that the inoculated EIAV(PV) quasispecies were all represented during the first febrile episode, but entirely replaced at the time of the second febrile episode, and that new predominant quasispecies were associated with each subsequent cycle of disease. One of the more surprising results was the in vivo generation of large deletion (up to 15 amino acids) in the principal neutralizing domain (PND) of gp90 during the third febrile episode. This deletion did not alter the competence for in vitro replication as shown by the analysis of a env chimeric clone with a partially deleted PND and did not altered the fitness of the virus in vivo, since this partially deleted envelope became the major population during the fourth febrile episode. Finally, we showed that the amino acid mutations were not randomly distributed but delineated eight variables regions, V1 to V8, with V3 containing the PND region. These studies provide the first detailed description of the evolution of EIAV genomic quasispecies during persistent infection and reveal new insights into the genetics and potential mechanisms of lentivirus genomic variation.

Amino Acid Sequence↗

RT-PCR detection of lentiviruses in milk or mammary secretions of sheep or goats from infected flocks.

In this study we evaluated a reverse transcriptase polymerase chain reaction (RT-PCR) technique for detecting lentiviral infection in milk or mammary secretions from small ruminants. Initial observations on seven goats infected with cloned caprine arthritis-encephalitis virus (CAEV) showed that RT-PCR on milk cells is as reliable as coculture for detecting viral infection, and is quicker and simpler. With a suitable choice of redundant primers followed by a semi-nested amplification, it proved possible to detect the virus in milk samples from naturally infected French sheep (8/8) or goats (9/9), and viral sub-groups could be identified by hybridization with discriminatory probes. All seropositive animals gave positive amplifications, as did one seronegative goat from a contaminated herd, suggesting greater sensitivity for RT-PCR. None of eight goats from a long-established seronegative herd ever gave a positive RT-PCR amplification. This technique provides a simple means for rapidly identifying potentially infectious animals and for epidemiological investigations, as long as the primers are selected according to the genetic structure of the local viral population.

Animals↗

[Treatment of common bile duct lithiasis: first-line endoscopic sphincterotomy and celioscopic cholecystectomy].

The aim of this study was to assess retrograd cholangiogram findings and first-line endoscopic sphincterotomy followed by laparoscopic cholecystectomy for the treatment of main bile duct lithiasis. Clinical, biological and echographic criteria predictive of main bile duct lithiasis were observed in 125 patients (32 men, 93 women, mean age 44.2 years) who underwent retrograde cholangiography. Results suggested lithiasis of the main bile duct in 105 case (87.5%) and were confirmed at endoscopic sphincterotomy in 99. There were no deaths; four complications occurred (3 moderate cases of pancreatitis, 1 cholecystitis). Conversion was required in 11.6%, usually because of difficulties in dissecting. No residual lithiasis was observed. Mean duration of hospitalization was 11.4 days. This sequential treatment scheme for main bile duct lithiasis appears to be effective, minimally invasive and safe.

Adult↗