Biomedical subjects

B Picard

Publications and source records attributed to B Picard.

At least 55 records · Page 3Linked to original sources

PubMed1997

Regional variations of muscle fibre characteristic in m. semitendinosus of growing cattle.

The objectives of the present study were to elaborate an intra-muscular profile of metabolic enzyme equipment, contractile and morphometric features along the longitudinal axis of m. semitendinosus at various ages throughout the growth phase. Thirty-seven male Montbéliard cattle, about half of them castrated, were representatively allocated to various slaughter dates, scheduled at 4, 8, 12 and 16 months of age. Samples were collected from proximal, medial and distal locations of m. semitendinosus. Isocitrate dehydrogenase (aerobic metabolism) and lactate dehydrogenase (anaerobic metabolism) were measured spectrophotometrically. Contractile muscle type was classified by quantification of myosin heavy chain I isoform proportion using the sensitive enzyme-linked immunosorbent assay. Mean muscle fibre area was obtained on histologically-stained cross-sections utilizing an image analysis system. Our results indicate the existence of a regular intra-muscular pattern of muscle fibre traits along the length of m. semitendinosus, with decreasing glycolytic activities and concomitantly an increase in oxidative capacity towards the distal extremity. The metabolic characteristics were in good agreement with decreasing cross-sectional muscle fibre areas and the slow myosin heavy chain I isoform proportion becoming gradually more abundant from proximal to distal regions of the muscle. Moreover, the observed gradient was found to be closely related to age and diminished with advanced physiological maturity. At the final slaughter age (16 months) no differences among the distinct portions were detected, m. semitendinosus was longitudinally homogeneous in all the characteristics studied.

Aging↗

PubMed1997

Weaning marginally affects glucose transporter (GLUT4) expression in calf muscles and adipose tissues.

The nutritional regulation of glucose transporter GLUT4 was studied in eight muscles and four adipose tissues from two groups of preruminant (PR) or ruminant (R) calves of similar age (170 d), empty body weight (194 kg) at slaughter, and level of net energy intake from birth onwards. Isocitrate dehydrogenase (EC 1.1.1.41) activity in muscles was not different between PR and R except in masseter muscle from the cheek (+71% in R; P < 0.003), which becomes almost constantly active at weaning for food chewing. Basal and maximally-insulin-stimulated glucose transport rate (GTR) per g tissue wet weight in rectus abdominis muscle were significantly higher in R calves (+31 and 41% respectively; P < 0.05). GLUT4 protein contents did not differ in muscles from PR and R except in masseter (+74% in R; P < 0.05) indicating that the increased GTR in rectus abdominis cannot be accounted for by an enhanced GLUT4 expression. GLUT4 mRNA levels did not differ between the two groups of animals in all muscles suggesting a regulation of GLUT4 at the protein level in masseter. GLUT4 number expressed on a per cell basis was lower in adipose tissue from R calves (-39%; P < 0.05) and higher in internal than in peripheral adipose tissues. In summary, the regulation of GLUT4 in calves at weaning differs markedly from that previously described in rodents (for review, see Girard et al. 1992). Furthermore, significant inter-individual variations were shown for metabolic activities in muscle and for biochemical variables in adipose tissue.

Abdomen↗

PubMed1997

An epidemiological study of Serratia marcescens isolates from nosocomial infections by enzyme electrophoresis.

Serratia marcescens isolates from 164 patients with suspected nosocomial infection in several hospitals in the greater Paris region were investigated by analysis of the electrophoretically demonstrable allelic variations of gene loci coding for five esterases and five other enzymes. All the loci were polymorphic and the mean number of alleles per locus was 6.1. A total of 72 distinctive electrophoretic types (ETs) representing multilocus genotypes was distinguished. The isolates were divided into two groups according to their resistance to antibiotics: 82 multiresistant isolates (MRI) and 82 relatively susceptible isolates (RSI). Seventy-two MRI (88%) were in four genetically related ETs: ET1, ET2, ET8 and ET9; ET1 was found in 48 isolates, whereas the remaining MRI were in 10 ETs, and all RSI in 61 ETs. Three ETs contained both MRI and RSI. The mean coefficients of genetic diversity for the 10 enzyme loci among ETs and isolates were smaller for MRI than for RSI, while the modal ET of MRI resembled that of RSI. The epidemiological significance of isolates varied according to their ET. Thus, isolates belonging to ET1, ET2 and ET8 were responsible for outbreaks or for sporadic infections, whereas isolates of other ETs were responsible for only sporadic infections. The temporal distribution of ET1 isolates among hospitals identified seven outbreaks in seven clinical departments.

Alleles↗

PubMed1996

Mechanisms of the spread of penicillin resistance in Streptococcus pneumoniae strains causing meningitis in children in France.

The molecular epidemiology of penicillin-resistant Streptococcus pneumoniae strains causing meningitis in children was studied in France. Typing procedures included analysis of total DNA polymorphism by random amplification of polymorphic DNA, restriction fragment length polymorphism (RFLP) analysis of the ribosomal RNA gene regions, and pulsed-field gel electrophoresis. Penicillin-binding protein (PBP) genes pbp2b and pbp2x were studied by RFLP analysis and DNA sequencing in selected cases. Statistical analysis of the data by factorial analysis of correspondence established that the emergence of penicillin-resistant pneumococci in this pathology is the result of the spread of two highly resistant closely related clusters and a cluster of serotype 23 strains with an intermediate level of resistance, the spread of genes confering high resistance to penicillin between the two highly resistant clusters, and complex genetic events involving the pbp genes in a heterogeneous population of strains leading to an intermediate level of resistance.

Base Sequence↗

PubMed1996

Analysis of genetic relationships among strains of groups A, C and G streptococci by random amplified polymorphic DNA analysis.

Random amplified polymorphic DNA (RAPD) analysis was evaluated for its capacity to distinguish species and strains within species of groups A, C and G streptococci. The 99 strains tested, previously typed by multilocus enzyme electrophoresis (MLEE), included 41 group A streptococci (Streptococcus pyogenes), 25 group G Streptococcus spp. (GGS), seven S. dysgalactiae, 11 S. equisimilis, four S. canis, three S. equi and eight S. zooepidemicus. The combined data obtained with three single primers distinguished 82 types. RAPD analysis provided taxonomic results that were in general agreement with previous species classification based on DNA-DNA homology and MLEE. The intraspecies typing efficiency of the technique was significantly improved by the parallel use of several primers. RAPD analysis had greater discriminatory power than MLEE for GAS and GGS. There was not total agreement between the two techniques as RAPD distinguished strains with identical electrophoretic types, whereas MLEE differentiated strains with identical PCR types. RAPD analysis did not distinguish all GAS strains with different biotypes and its already high discriminatory power was further enhanced by concomitant biotyping.

Animals↗

PubMed1996

Molecular epidemiological tools for Salmonella Dublin typing.

A total of 32 strains of Salmonella Dublin recovered from cattle were differentiated by electrophoretic typing of their esterases (zymotyping), restriction fragment length polymorphism of ribosomal DNA (ribotyping), arbitrarily primed PCR (AP-PCR) using five primers, PCR based on repetitive extragenic palindromic sequences (REP-PCR) and PCR based on enterobacterial repetitive intergenic consensus sequences (ERIC-PCR). ERIC-PCR and REP-PCR each gave one type, zymotyping gave three, AP-PCR gave five and ribotyping gave seven types. Combination of ribotyping and AP-PCR produced a total of 11 types, whereas 14 different types were obtained by all five methods. Thus a combination of several methods enhanced the discrimination of cattle-adapted strains among the genotypically homogeneous serovar Salmonella Dublin.

Animals↗

PubMed1996

Multicenter evaluating of a commercially available PCR assay for diagnosing enterovirus infection in a panel of cerebrospinal fluid specimens.

Thirteen laboratories participated in blind tests of a panel of 20 coded cerebrospinal fluid specimens (7 uninfected samples, 3 samples infected with 1 50% tissue culture infective dose [TCID50]/0.1 ml [nonenterovirus strains], and 10 samples infected with 10, 1, or 0.1 TCID50/0.1 ml [three different enterovirus serotypes]) on the Amplicor enterovirus PCR assay (Roche Diagnostic Systems). The panel was also evaluated by in-house PCR (two nested-PCR and three one-step PCR assay) or tissue culture (eight laboratories). The viral load was shown to influence greatly the sensitivity of the assay. The average sensitivity of the Amplicor test ranged from 67 to 98% for viral titers of 1 to 10 TCID50/0.1 ml, respectively; titers of 0.1 TCID50/0.1 ml resulted in a sensitivity of only 16%. The overall specificity of the Amplicor test was 98%. The Amplicor assay compared favorably to the five in-house PCR tests (no significant difference in either sensitivity or specificity) and was much more sensitive than tissue culture (P < 0.001), even for high viral loads. It was easy to perform, rapid (about 6 h), well-standardized, and appeared to be suitable for the diagnosis of enterovirus meningitis on a routine basis in laboratories trained in molecular biology techniques.

Base Sequence↗

PubMed1995

Comparison of the foetal development of muscle in normal and double-muscled cattle.

Muscle differentiation was studied in foetal Semitendinosus muscle from normal cattle and those with the 'culard' gene of muscular hypertrophy sampled at 90, 130, 170 and 210 days of foetal life. The different fibre types were characterized by immunohistochemistry with antibodies specific to different isoforms of myosin heavy chains. The isoforms were separated by electrophoresis, identified by immunoblotting and quantified by ELISA. In double-muscled animals, there was a slower rate of differentiation in the first generation of cells, most markedly apparent at 90 days. At 130 days, differentiation was retarded mainly in the second generation, while at 170 days there was no longer any difference between the two animal types in the differentiation of first generation cells, which were totally slow in both. At the same stage however, type IIC fibres in double-muscled animals were much slower in appearing and continued to be so at 210 days, albeit to a lesser extent. These findings show that differentiation of the muscle fibres occurs at a slower rate in double-muscled foetuses particularly during the first two-thirds of foetal life.

Animals↗

PubMed1995

Sex in Escherichia coli does not disrupt the clonal structure of the population: evidence from random amplified polymorphic DNA and restriction-fragment-length polymorphism.

Analysis of the Escherichia coli population by multilocus enzyme electrophoresis (MLEE) has established its clonal organization, but there is increasing evidence that horizontal DNA transfer occurs in E. coli. We have assessed the genetic structure of the species E. coli and determined the extent to which recombination can affect the clonal structure of bacteria. A panel of 72 E. coli strains from the ECOR collection was characterized by random amplified polymorphic DNA (RAPD) and restriction-fragment-length polymorphism (RFLP) of the ribosomal RNA gene (rrn) regions. These strains have been characterized by MLEE and are assumed to reflect the range of genotypic variation in the species as a whole. Statistical analysis, including factorial analysis of correspondence (FAC) and hierarchical classifications, established that the data obtained with the three genetic markers are mutually corroborative, thus providing compelling evidence that horizontal transfer does not disrupt the clonal organization of the population. However, there is a gradient of correlation between the different classifications which ranges from the highly clonal structure of B2 group strains causing extraintestinal infections in humans to the less-stringent structure of B1 group strains that came mainly from nonprimate mammals. This group (B1) appears to be the framework from which the remaining non-A group strains have emerged. These results indicate that RAPD analysis is well suited to intraspecies characterization of E. coli. Lastly, treating the RAPD data by FAC allowed description of subgroup-specific DNA fragments which can be used, in a strategy comparable to positional cloning, to isolate virulence genes.

Base Sequence↗

PubMed1995

Activity of beta-lactamase inhibitor combinations on Escherichia coli isolates exhibiting various patterns of resistance to beta-lactam agents.

The efficacy of the clinically available beta-lactam/beta-lactamase inhibitor combinations (amoxicillin/clavulanic acid (CA), ticarcillin/CA, amoxicillin/sulbactam, and piperacillin/tazobactam) was evaluated on 300 amoxicillin-resistant Escherichia coli isolates having the main patterns of beta-lactam resistance. The patterns, which reflect the production of various beta-lactamase enzymes, were analyzed by a principal component analysis of susceptibility to 11 beta-lactam antibiotics or beta-lactam/beta-lactamase inhibitor combinations. Sixty-two percent of strains were not very susceptible to penicillins, cephalothin, or any beta-lactam/beta-lactamase inhibitor combinations except for piperacillin/tazobactam; these strains may represent high-level broad-spectrum beta-lactamase (so-called penicillinase) production phenotype or inhibitor-resistant TEM-like enzyme production phenotype. Of the strains, 14.7% were resistant to amoxicillin and ticarcillin compatible with low-level broad-spectrum beta-lactamase production phenotype; 5.7% were cefoxitin resistant and were postulated to present a high-level cephalosporinase production phenotype; and 2.6% were resistant to cephalothin only, attributable to a low-level cephalosporinase production phenotype. Three percent of strains were intermediate or resistant to cefotaxime and may produce an extended-spectrum beta-lactamase, and the remaining strains (12 %), resistant to all tested antibiotics except for cefotaxime and piperacillin/tazobactam, were hypothesized to produce both broad-spectrum beta-lactamase plus cephalosporinase. The minimal inhibitory concentration (MIC) for these phenotype patterns indicated that combinations of CA plus amoxicillin or ticarcillin, or sulbactam plus amoxicillin, restored the activity of penicillins against phenotype 1 strains, whereas these combinations remained inactive against the other phenotype strains. Piperacillin plus tazobactam showed the best in vitro effect against the strains of all resistance phenotypes.

Amoxicillin↗

PubMed1995

Study of the influence of age and weaning on the contractile and metabolic characteristics of bovine muscle.

Weaning is an interesting period for the study of the nutritional regulation of muscle energy metabolism, since during this stage the nature of the substrates supplied to the muscle and their energy balance are profoundly changed. The aim of this study was to determine the effect of these modifications on the contractile and metabolic characteristics of bovine muscle. Two similar groups of 7 male Montbéliard calves were used with the same age and weight, and with the same energy intake. One group consisted of milk-fed calves, the other of weaned animals. The latter were progressively weaned over a period between 107 and 128 d. The average age at slaughter in the 2 groups was 170 d. Biopsy specimens of semitendinosus (ST) muscle were taken at the ages of 66 d, 94 d (before the beginning of weaning) and 136 d (at the end of weaning) to follow the evolution of muscle characteristics. Samples of longissimus thoracis (LT) muscle were taken 24 h after slaughter and used to study the changes in protein and DNA content. The proportion and area of the different types of fiber, I (slow, oxidative), IIA (fast, oxido-glycolytic), IIB (fast, glycolytic) and IIC (fast/slow, oxidoglycolytic) were measured by immunohistochemistry and image analysis. The metabolism of the muscles was determined by studying isocitrate dehydrogenase (ICDH, oxidative) and lactate dehydrogenase (LDH, glycolytic) activity. The results obtained between 2 and 6 months of life showed an overall increase in the area of the fibers (I, IIA, IIB and IIC) and a conversion of type IIA fibers into type IIB accompanied by a shift in the energy metabolism towards a glycolytic type. Weaning caused temporary stress, whose main consequences were to decrease overall muscle fiber area and the percentage of type IIB fibers, and increase the proportion of type IIC fibers in weaned animals. These effects may have been due to the nutritional and behavioral disturbances that accompany weaning, because 42 d after the end of weaning there was no difference in the size of ST and LT fibers between the 2 groups whereas the proportion of type IIA fibers was still higher in weaned animals.

Aging↗

PubMed1995

Enzyme-linked immunosorbent assay for myosin heavy chains in the horse.

The content in slow and fast myosin heavy chains (MHC 1 and MHC 2) of 5 equine muscles was determined using an enzyme-linked immunosorbent assay. The results obtained with this immunoenzymatic method were compared with complementary techniques: electrophoresis and immunohistochemistry. Slices of masseter, diaphragm, tensor faciae latae, semitendinosus and cutaneus trunci were obtained from a 12-year-old saddle horse after slaughter. Muscular proteins were specifically extracted to be analysed by ELISA. The technique used 2 complimentary monoclonal antibodies (MAb). MAb 1 was prepared from a human atrium specimen that reacted specifically against MHC 1. Mab 2 was prepared from myosin of rabbit psoas muscle and reacted against MHC 2. The masseter muscle contained solely MHC 1 (100%) and this was confirmed by electrophoresis and immunohistochemistry. By contrast, the cutaneus trunci was very poor in MHC 1 (1.3%) and was entirely composed of MHC 2 (98.7%) which was confirmed by the other techniques. The diaphragm, tensor fasciae latae and semitendinosus contained 89, 40 and 2% of MHC 1, respectively. It was concluded that this ELISA method made it possible to measure a wide range of MHC contents in equine muscles with a good reproducibility. The results were consistent with those of the other fibre typing techniques. Moreover, this immunoenzymatic method is less time consuming than histological techniques and therefore offers new perspectives for muscle fibre typing in the horse.

Animals↗

PubMed1995

Identification and typing of pyogenic streptococci by enzyme electrophoretic polymorphism.

Polyacrylamide-agarose gel electrophoresis was used to study polymorphism of lactate dehydrogenase (LDH), nucleoside phosphorylase (NSP), phosphoglucose isomerase (PGI), hydroxybutyrate dehydrogenase (HBD), adenylate kinase (ADK) and esterases of 44 strains of Streptococcus pyogenes, 25 group G streptococcal strains, 11 "S. equisimilis" strains, seven S. dysgalactiae strains, four S. canis strains, three S. equi strains and seven S. zooepidemicus strains. Analysis of LDH, NSP, PGI, HBD and ADK provided valuable interspecies differentiation, by showing that four groups of strains corresponded to the four known DNA homology groups. Esterases showed greater intraspecies variation than the other enzymes. The combined analysis of the six enzymes indicated 31 zymotypes among S. pyogenes, 14 in group G streptococci and 11 in "S. equisimilis" strains. This was shown to be an effective technique for typing pyogenic streptococci.

Adenylate Kinase↗

PubMed1994

Genomic fingerprinting of Yersinia enterocolitica species by degenerate oligonucleotide-primed polymerase chain reaction.

A novel version of the polymerase chain reaction (PCR) termed degenerate oligonucleotide-primed PCR (DOP-PCR) was used to fingerprint the bacterial genome of Yersinia enterocolitica species. Eight well-characterized reference strains of Y. enterocolitica (ribotype, serotype, biotype and zymotype) were examined. Optimal experimental conditions for reproducibility, sensitivity and specificity were obtained using a single DOP-primer. All the strains gave distinct DOP-PCR profiles composed of 12 to 19 fragments with sizes from 200 to 1.500 bp. Thus, the DOP-PCR, which has so far been used to fingerprint human, mouse, fruitfly and plant DNA, is also well-suited to bacterial DNA.

Base Sequence↗

PubMed1994

A simple procedure to differentiate ailA and ailNA gene variants among human pathogenic Yersinia enterocolitica strains.

The identification and differentiation of the two variants of the ail gene, ailA from the more virulent American serotypes (08, 013a, 13b, 018, 020, 021) and ailNA from the less virulent non-American serotypes (03, 04, 05, 06, 09, 027 and 07, 8) was studied in a panel of 32 Yersinia enterocolitica human pathogenic isolates. A 444 bp fragment corresponding to the ail gene was amplified using a PCR procedure in all tested strains. Subsequent digestion of the PCR product by Rsal and by HaeIII endonucleases, provide electrophoretic patterns that clearly discriminate ailA and ailNA variants. This non-radioactive and reliable procedure allows large clinical and epidemiological studies, and could be proposed to survey the spread of virulent clones.

Base Sequence↗

PubMed1994

Comparison of the foetal development of fibre types in four bovine muscles.

The pattern of expression of different types of myosin heavy chains and the development of different generations of muscle cells during foetal life were studied in four bovine muscles with widely varying characteristics, the Masseter, Longissimus thoracis, Cutaneus trunci and Diaphragma. Different complementary techniques were performed: immunocytochemistry, electrophoresis, immunoblotting and ELISA. Monoclonal antibodies against different myosin heavy chain isoforms were used. The results confirmed the existence of at least two generations of cells during foetal development in cattle. A first generation, which appeared at a very early stage, gave rise to adult type I fibres. A second generation, made up of different cell populations, gave rise to adult fast type IIA and IIB fibres, and to type IIC. In the slow muscles, it also seemed to give rise to type I fibres. The beginning of myogenesis was characterized in the different cell generations by the expression of transitory myosin forms that are not found in the adult. Type 1 myosin heavy chain was observed from 90 days whereas the fast types, 2a and 2b, were present from 210 to 230 days, at which stage the foetal form disappeared. Muscles that have greatly different contractile characteristics in the adult exhibit also different profiles of differentiation: the Diaphragma was the first to develop, followed by Cutaneus trunci, Longissimus thoracis and Masseter.

Animals↗

PubMed1994

Correlation between esterase electrophoretic polymorphism and virulence-associated traits in extra-intestinal invasive strains of Escherichia coli.

The electrophoretic variations of carboxylesterase B and of esterases A, C and I, the presence of mannose resistant haemagglutinin, alpha-haemolysin, cytotoxic necrotizing factor type 1 (CNF1) and certain O antigens were compared in 150 strains of Escherichia coli responsible for extra-intestinal infections. Electrophoretic mobilities of outer membrane proteins (OMP) were also studied for strains belonging to O4, O6, O7, O8 and O75 serogroups. Fast migrating allozymes of carboxylesterase B (pattern B1) were correlated with slow migrating allozymes of esterase C, serogroups O7 and O8, lack of virulence factor, and particular OMP patterns, whereas slow migrating allozymes of carboxylesterase B (pattern B2) were correlated with fast migrating allozymes of esterase C, serogroups O2, O4, O6, O18 and O75, virulence factor production, and distinct OMP patterns. Allozymes of esterases A and I were not clearly correlated with the distribution of virulence factors. The pattern B2 was more strongly associated with CNF1 than with alpha-haemolysin and mannose resistant haemagglutinin. These results substantiate the view that the electrophoretic pattern B2 of carboxylesterase B identified most of the highly pathogenic strains implicated in extra-intestinal infection of humans.

Bacteremia↗

PubMed1994

O, K, and H antigens predict virulence factors, carboxylesterase B pattern, antimicrobial resistance, and host compromise among Escherichia coli strains causing urosepsis.

The O:K:H serotypes of 75 Escherichia coli blood isolates from patients with urosepsis were compared for the presence and expression of determinants for P fimbriae, hemolysin, and aerobactin; antimicrobial resistance; the carboxylesterase B phenotype; and associated compromising host conditions. O groups, K types, and O:K:H serotypes previously associated with urovirulence accounted for 69%, 60%, and 31% of the population, respectively. Chromosomal determinants for P fimbriae, hemolysin, and aerobactin were present in combination more commonly among strains belonging to urovirulence-associated O groups, K types, and O:K:H serotypes. Similarly, antimicrobial resistance was strikingly less prevalent, the B2 carboxylesterase phenotype more common, and associated host compromise less common among such strains. These data demonstrate that the O groups, K types, and O:K:H serotypes traditionally associated with urovirulence are prominent among E. coli strains causing urosepsis, in which they are associated with presence and expression of multiple chromosomal virulence factor determinants, susceptibility to antimicrobial agents, the B2 carboxylesterase phenotype, and noncompromised hosts.

Antigens, Bacterial↗