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J L Pons

Publications and source records attributed to J L Pons.

15 recordsLinked to original sources

Clonal and temporal patterns of nasopharyngeal penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae stains in children attending a day care center.

The nasopharyngeal carriage of penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae (PSSp and PRSp, respectively) was analyzed in 116 children attending a day care center in Rouen, France, by three observation periods in November, January, and March of the winter of 1993 to 1994. The carriage rate of S. pneumoniae was found to be 47.7, 47.3, and 49.6% at each different observation period, and PRSp accounted for 42.2, 40.3, and 40.6% of pneumococcal isolates, respectively. The 52 isolates recovered in November were distributed in 34 electrophoretic types (ETs) by multilocus enzyme electrophoresis; 15 PRSp isolates, all of serotype 23F, belonged to a clonal complex of five ETs, representing the dominant population of PRSp in November. The temporal pattern of S. pneumoniae carriage was studied in 17 children who were colonized at the three periods by multilocus enzyme analysis of their isolates. The PSSp isolated, exhibiting distinct ETs, were transient only among these day care attendees. In contrast, most of the PRSp isolated in January and March belonged to the clonal complex. Thus, this PRSp population was resident in the day care center throughout the study period.

Carrier State

Evaluation of the Epsilometer test (E test) for testing the susceptibility of coagulase-negative staphylococci to teicoplanin.

The antimicrobial susceptibilities of 118 clinical isolates of coagulase-negative staphylococci to teicoplanin were determined by disc diffusion and the Epsilometer test (E test) and the results were compared with the MICs determined by the agar dilution method of the National Committee for Clinical Laboratory Standards (NCCLS). There was a poor correlation of r = 0.5 between the zone diameters of inhibition and agar dilution MICs and 10 and four of the 11 isolates for which the MICs were > or = 32 mg/L were misclassified as susceptible by the disc test after applying the interpretative criteria of the NCCLS and the Comité de l'Antibiogramme de la Société Française de Microbiologie (CASFM), respectively. The E test tended to result in MICs that were lower than those determined by agar dilution and only 66% of MIC were within +/- 1 log2 dilution of each other. Only one of 11 resistant strains was detected by the E test and, although there was no false resistance, six resistant strains were misclassified as susceptible after applying the criteria of the NCCLS as were four such isolates when the criteria of the CASFM were employed, probably as a result of using too light an inoculum. Disc diffusion is not a reliable means of determining the susceptibility of coagulase-negative staphylococci but might be replaced by the E-test provided that discrepant results can be resolved by using a denser inoculum.

Anti-Bacterial Agents

Methicillin-resistant Staphylococcus aureus subtyping: interest of combined antibiotyping and esterase electrophoretic typing.

Ninety-four methicillin-resistant Staphylococcus aureus isolates (MRSA) were characterized by means of two typing methods, antibiotyping and esterase electrophoretic typing. Antibiotyping, recorded on the basis of susceptibility testing of 13 antimicrobial agents, allowed the description of 18 antibiotypes, four of which comprised 30, 14, 14 and 12 strains respectively. Esterase electrophoretic typing, based on esterase activity against seven synthetic substrates after polyacrylamide-agarose gel electrophoresis, led to the description of 12 electrophoretic types, two of which were predominant with 60 and 20 strains respectively. The combined use of both typing methods yielded 32 combinations, three of which were predominant with 21, 12 and 11 strains respectively. A good differentiation of strains was achieved, particularly when the antibiotype was correlated to the electrophoretic type. Thus, the combination of antibiotyping with esterase electrophoretic typing may be proposed as a well-suited method for the characterization of MRSA strains.

Bacterial Typing Techniques

[Outbreak of infections of Klebsiella pneumoniae with extended spectrum beta-lactamase in a hospital unit with a long and medium stay].

Fourteen extended-spectrum beta-lactamase (ES beta la) Klebsiella pneumoniae strains were isolated from 14 inpatients between February 1993 and February 1994, in a medium- and long-stay neurological unit. For this reason an epidemiological study was begun, based on strain typing and examination of patient files. Strain typing was carried out by two methods (i) the analysis of antibiotic resistance, showing 7 different antibiotypes among the 14 strains studied, (ii) the analysis of esterase and dehydrogenase electrophoretic polymorphism in polyacrylamide-agarose gel. The method was checked by analysing 11 Klebsiella pneumoniae strains with wild phenotype for beta-lactam antibiotics, which were isolated during the same period in the same unit. Simultaneously 6 other strains isolated during the same period in some other units of the hospital were analysed. Nine electrophoretic types were found among the 31 strains (wild and ES beta las). The analysis of the results showed that 8 isolates of the group of 14 ES beta las had the same antibiotype and electrophoretic type. This demonstrates that one epidemic strain was responsible for two outbreaks, the first one in April and the second one in August-September. A case control investigation was carried out to define the risk factors of infection. Files were examined for the 14 infected inpatients and for 20 control inpatients from the same unit during the same period. Statistical analysis was performed with Epi Info software 5 (CDC Atlanta). Length of stay, dependence and malnutrition levels, and urinary sphincter disfunction were the most significant risk factors.

Anti-Bacterial Agents

Multilocus enzyme typing of human and animal strains of Clostridium perfringens.

Multilocus enzyme electrophoresis was developed to evaluate the genetic diversity of 71 human strains and 17 animal strains of Clostridium perfringens. Crude protein extracts, obtained by sonication of washed bacteria, were analyzed by polyacrylamide-agarose gel electrophoresis to characterize electrophoretic mobility variants of seven enzymes (esterase, glutamate dehydrogenase, glutamic-oxaloacetic transaminase, nucleoside phosphorylase, phosphoglucose isomerase, phosphoglucomutase, threonine dehydrogenase). Genetic diversity of the enzyme loci ranged from 0.340 to 0.813. Sixty-nine electrophoretic types were described among the 88 strains tested and the index of discrimination was 0.994. All strains were typable, and epidemiological relationships between isolates could be established. This method showed a fair correlation with esterase electrophoretic typing based on hydrolytic and electrophoretic polymorphism of esterases. This work demonstrates that multilocus enzyme polymorphism is a reliable and discriminant marker of genetic diversity of strains of C. perfringens.

Animals

Clonal outbreaks of extended-spectrum beta-lactamase-producing strains of Klebsiella pneumoniae demonstrated by antibiotic susceptibility testing, beta-lactamase typing, and multilocus enzyme electrophoresis.

Nineteen extended-spectrum beta-lactamase (ESBla)-producing Klebsiella pneumoniae isolates from Rouen Hospital were investigated for their implication in nosocomial outbreaks: in addition to antibiotic susceptibility testing, the ESBlas were characterized by isoelectric focusing, and the genetic relationships between the strains were analyzed by multilocus enzyme electrophoresis using a combined polyacrylamide electrophoresis-electrophoretic transfer technique. Four isoelectric focusing beta-lactamase patterns and 11 enzyme electrophoretic types (ETs) among the strains tested were described. Three strains isolated in the same neurological unit over a 7-day period exhibited an SHV 3 beta-lactamase (pI 7.0) and were assigned to a common ET. Three of five strains isolated from patients in a rehabilitation center over a 6-week period harbored an SHV 4 beta-lactamase (pI 7.8) and exhibited the same ET. These results differentiate nosocomial transmission from sporadic cases and provide evidence that multilocus enzyme electrophoresis is a potential tool for studying genetic relationships between strains harboring a common ESBla.

Cross Infection

Esterase electrophoretic polymorphism of human and animal strains of Clostridium perfringens.

Esterase electrophoretic polymorphism in human and animal strains of Clostridium perfringens was studied by using polyacrylamide-agarose gel electrophoresis. Five types of esterases, designated E-I to E-V and defined by their hydrolytic specificities toward five synthetic substrates, were found in protein extracts of bacteria grown without glucose (glucose-containing media allowed only the expression of esterase E-I). Mobility variants of esterase E-I, which hydrolyzes alpha- and beta-naphthyl acetates and butyrates, were used as a basis for the distribution of strains into 11 zymogroups. When all five types of esterases and their electrophoretic variants were considered, 77 electrophoretic types (ETs) could be described for the 89 strains tested. Animal strains did not constitute a distinctive subpopulation, as revealed by their distribution in the zymogroups and by clustering analysis. Statistical analysis also emphasized the importance of esterase E-IV (which hydrolyzes only naphthyl acetates) and esterase E-V (which hydrolyzes only alpha-naphthyl acetate) in clustering by the relatedness of the ETs. ETs allowed the epidemiological characterization of stool isolates recovered from elderly inpatient residents and from adolescent chronic-care psychiatric patients. These results indicate that esterase electrophoretic typing may be a marker for epidemiological and ecological analyses.

Animals

Use of electrophoretic polymorphisms of esterases for differentiation of Clostridium argentinense strains.

Esterase electrophoresis was used to study 10 strains of Clostridium argentinense, including 7 toxigenic and 3 nontoxigenic strains. On the basis of the electrophoretic mobilities and hydrolytic specificities toward five synthetic substrates, different esterase profiles could be defined for almost all strains, revealing the heterogeneity of bacterial clones. Therefore, electrophoretic polymorphism of esterases can be used for differentiation of C. argentinense in population genetic or epidemiological studies.

Bacterial Typing Techniques

Comparison of MRC-5 and continuous cell lines for detection of cytomegalovirus in centrifugation cultures.

Continuous cell lines were assessed for use for rapid human cytomegalovirus (HCMV) detection procedures combining tissue culture, centrifugation, and immediate early antigen (IEA) immunostaining. Human cells (MRC-5 embryonic fibroblasts, U-373MG astrocytoma cells, differentiated teratocarcinoma (Tera-2) cells), murine cells (BALB/c-3T3 and Y-1 cells), BHK21 hamster cells, and mink lung (ML) cells were first inoculated with HCMV laboratory strain. IEA synthesizing cells were detected by immunoperoxidase assay using a monoclonal antibody. ML cells and differentiated Tera-2 cells exhibited more positive cells than MRC-5 cells. BHK21, and MRC-5 cells were equivalent in sensitivity whereas U-373MG, BALB/c-3T3, and Y-1 cells had only reduced IEA positive cells. When 63 urine specimens were inoculated onto MRC-5, ML and differentiated Tera-2 cells, 20 (31.7%) were positive in MRC-5 cells versus 18 (28.5%) in ML or Tera-2 cells. Moreover, greater numbers of infected cells were detected in MRC-5 cells than in these two cell lines. MRC-5 cells were superior for detection of HCMV in clinical samples by centrifugation cultures.

Animals

Evaluation of antimicrobial interactions between chlorhexidine, quaternary ammonium compounds, preservatives and excipients.

The antimicrobial interactions of 49 combinations of chlorhexidine, quaternary ammonium compounds, preservatives and excipients were evaluated by the method of Berenbaum and the checkerboard titration method, with Staphylococcus aureus CIP 53154 and Escherichia coli CIP 54127 as test strains. MIC determinations were carried out as a preliminary step, and relative growth intensity was used to describe the bacteriostatic activity of surface-active agents (Amonyl 380 BA, Amonyl 671 SB). In the study of combinations, results were interpreted with Fractional Inhibitory Concentration indexes and represented by isobolograms. A fair correlation was shown between the method of Berenbaum and the checkerboard titration method. Combinations between chlorhexidine, cetrimonium bromide and benzalkonium chloride were synergistic or additive; combinations of antiseptics and preservatives were generally not antagonistic. The methods were also well adapted to the study of interactions involving surface-active agents, a critical problem in the formulation of topical antimicrobial agents.

Anti-Infective Agents, Local

Standardized method for evaluation of hand disinfection by surgical scrub formulations.

A standardized protocol for the evaluation of hand disinfection by surgical scrub formulations was applied to volunteers in a multicenter trial. Povidone iodine (PVI), chlorhexidine (CHX), and a nonmedicated soap (NMS) were tested. The scrubbing procedure involved three daily hand washings for five consecutive days; surviving bacteria were counted daily after being collected in a suitable neutralizing solution. Immediate efficacy (IE), cumulative efficacy (CE), and remanent effect (RE) were calculated by reference to the control hand. Statistical analyses of IE, CE, and RE showed significant differences among the three scrub formulations. IEs of PVI and CHX were equivalent and different from IE of NMS; CE and RE of CHX were higher than those of PVI and NMS. On the basis of the statistical analysis, the population size required for further studies aimed at detecting significant differences between surgical scrub formulations could be estimated.

Adult

[Leukoedema].

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Humans

Gas chromatographic-mass spectrometric analysis of volatile amines produced by several strains of Clostridium.

A gas chromatographic--mass spectrometric technique is proposed for the analysis of volatile amines which were isolated from Clostridium cultures by vacuum distillation and concentrated as hydrochloride salts. Headspace sampling after alkalinization of the salts under vacuum was the most suitable for subsequent gas chromatographic analysis. With ammonia-loaded helium as carrier gas, methylamines were separated on 4.8% PEG 2OM + 0.3% potassium hydroxide on Carbopack B, and other volatile amines on 28% Pennwalt 223 + 4% potassium hydroxide on Gas-Chrom R. Bacterial volatile amines (dimethylamine, trimethylamine, isobutylamine, 3-methylbutylamine, etc.) were detected with a flame-ionization detector and identified by gas chromatography--mass spectrometry in electron-impact and chemical ionization modes.

Amines

[Biosynthesis of toluene in Clostridium aerofoetidum strain WS].

Formation of toluene in growing cultures of Clostridium aerofoetidum strain WS was enhanced when the medium was supplemented with phenylacetic acid or with L-phenylalanine and L-methionine together. Evidence for the role of L-phenylalanine was shown by the detection of [2H2]-methyl[2,3,4,5,6-2H5]benzene ("heptadeuterotoluene") in growing cultures with L-[2',3',4',5',6'-2H5]phenyl[2,3-2H3]alanine and L-methionine.

Clostridium