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J Duval

Publications and source records attributed to J Duval.

At least 19 recordsLinked to original sources

New high-content disks for determination of high-level aminoglycoside resistance in clinical isolates of Enterococcus faecalis.

Disks impregnated with 500 and 1000 micrograms of streptomycin, 1000 micrograms of kanamycin and 250 and 500 micrograms of gentamicin were used for detection of high-level resistance to aminoglycosides in 120 clinical isolates of Enterococcus faecalis. Fifty-seven strains were highly resistant to streptomycin, 80 to kanamycin and 41 to gentamicin. Using disks containing 500 micrograms of streptomycin, 1000 micrograms of kanamycin and 500 micrograms of gentamicin strains resistant to high levels of these drugs (97.9%, 100% and 100%, respectively) were accurately detected. Better discrimination between high-level and low-level resistance was achieved with a 500 micrograms streptomycin or gentamicin disk. Zone-size breakpoints are proposed for detection of high-level resistance by disk diffusion.

Drug Resistance, Microbial

Saponins from stem bark of Petersianthus macrocarpus.

Two bioactive saponins were isolated from the stem bark of Petersianthus macrocarpus. Their structures were elucidated by chemical degradations and by a combination of 2D NMR techniques and by Californium plasma desorption mass spectrometry. They are 3-O-([beta-D-galactopyranosyl (1-->2)][beta-D-galactopyranosyl (1-->3)]- beta-D-glucuronopyranosyl)-21-O-[3-(3-tigloyloxynilic acid)-4-tigloyloxy- alpha-L-arabinopyranosyl] barringtogenol C and 3-O-([beta-D-galactopyranosyl (1-->2)][beta-D-galactopyranosyl (1-->3)]-beta-D-glucuronopyranosyl)-28-O-alpha-L-rhamnopyranosyl barringtogenol C-21-O-benzoate. The absolute configuration of nilic acid was determined by partial synthesis. 3,3'-Dimethoxy ellagic acid and 3,3'-dimethoxy-4-O-beta-D- glucopyranosyl ellagic acid were also isolated.

Carbohydrate Sequence

Direct estradiol down-regulation of secretogranin II and chromogranin A mRNA levels in rat pituitary cells.

Estradiol (E2) has been previously shown to negatively regulate, in vivo, the secretogranin (SgII) and chromogranin A (CgA) mRNA levels in the rat pituitary. Using cultured pituitary cell aggregates, experiments were undertaken to discriminate between direct or indirect effects of E2 on SgII and CgA levels. SgII, CgA and gonadotropin alpha- and beta-subunit mRNA levels were determined by Northern blotting. SgII and CgA protein levels were quantitated by Western blotting, and by immunoprecipitation of radioactive SgII after [35S]methionine labeling. Increasing concentrations of E2 (from 10(-12) M to 10(-8) M) in the culture medium promoted a decrease of SgII and CgA mRNA levels to 30% and 50% of the control, respectively, after 72.h treatment. By contrast, none of the gonadotropin subunit mRNAs exhibited changes in concentration. A 24 h treatment with 10(-8) M E2 was sufficient to promote such a decrease in SgII and CgA mRNAs. Quantitation of the proteins after Western blotting revealed that 10(-8) M E2 lowered by 30% in CgA content of aggregates (P < 0.05 vs. control) while SgII content remained unaffected. Moreover, quantitation of the newly synthesized SgII by immunoprecipitation of the 35S-labeled SgII gave evidence for a lack of E2 effect. These data demonstrate: (1) a direct effect of E2 on the pituitary cells to down-regulate SgII and CgA mRNA steady-state levels; (2) though contained within the same secretory granules, a distinct pathway for negative E2 regulation of the gonadotropins and both granins; and (3) a differential effect of E2 on cell SgII and CgA contents as was previously demonstrated in vivo.

Animals

Stimulation of LH release from cultured pituitary cells by saponins of Petersianthus macrocarpus: a permeabilizing effect.

Aqueous extracts from stem bark of Petersianthus macrocarpus contain substances exhibiting both estrogenic and antiestrogenic potency. Triterpenic saponins were identified and extracted as a bulk. Their action on the in vitro LH released by cultured rat pituitary cells was investigated. P. macrocarpus saponins stimulated the LH release in a dose-dependent manner (from 10 micrograms/ml to 300 micrograms/ml). When added simultaneously, saponins and LHRH exerted initial additive effects on LH release, demonstrating independent mechanisms of stimulation. If cells were pre-treated with saponins for 15 min, the amount of LH released under a subsequent LHRH stimulation was lowered, presumably due to a partial depletion of the cells in hormone (data not presented). However, the action of saponins on LH release did not appear specific since a general permeabilizing effect of the cell membrane was evidenced both by trypan blue exclusion and by analysis of the total protein output. When using low concentrations of saponins (10 micrograms/ml), scanning electron microscopy did not reveal any significant alteration of the cell structure, which explains why the cells remain responsive to LHRH after withdrawal of saponins. With higher concentrations (greater than 30 micrograms/ml), the same analytical studies evidenced numerous perforations of the cell membrane, with subsequent cell death. Two highly purified saponin species were tested on LH release by cultured cells; one of them (petersaponin I) appeared responsible for the observed biological effects in vitro. But as cells were shown to be efficiently protected from saponin effects by the presence of serum, it may be concluded that saponins of P. macrocarpus extracts are probably not candidate molecules promoting the in vivo estrogenic and antiestrogenic effects.

Animals

Resistance of enterococci to aminoglycosides and glycopeptides.

High-level resistance to aminoglycosides in enterococci often is mediated by aminoglycoside-modifying enzymes, and the corresponding genes generally are located on self-transferable plasmids. These enzymes are similar to those in staphylococci but differ from the modifying enzymes of gram-negative bacteria. Three classes of enzymes are distinguished, depending upon the reaction catalyzed. All but amikacin and netilmicin confer high-level resistance to the antibiotics that are modified in vitro. However, the synergistic activity of these last two antibiotics in combination with beta-lactam agents can be suppressed, as has always been found in relation to high-level resistance to the aminoglycosides. Acquisition of glycopeptide resistance by enterococci recently was reported. Strains of two phenotypes have been distinguished: those that are resistant to high levels of vancomycin and teicoplanin and those that are inducibly resistant to low levels of vancomycin and susceptible to teicoplanin. In strains of Enterococcus faecium highly resistant to glycopeptides, we have characterized plasmids ranging from 34 to 40 kilobases that are often self-transferable to other gram-positive organisms. The resistance gene vanA has been cloned, and its nucleotide sequence has been determined. Hybridization experiments showed that this resistance determinant is present in all of our enterococcal strains that are highly resistant to glycopeptides. The vanA gene is part of a cluster of plasmid genes responsible for synthesis of peptidoglycan precursors containing a depsipeptide instead of the usual D-alanyl-D-alanine terminus. Reduced affinity of glycopeptides to these precursors confers resistance to the antibiotics.

Aminoglycosides

A collaborative study of the in-vitro sensitivity to RP 59500 of bacteria isolated in seven hospitals in France.

The in-vitro activity of RP 59500 was determined against 1051 recent clinical bacterial isolates. The susceptibility to RP 59500 was determined with an agar dilution technique for all the isolates, while MICs and MBCs were determined for 82 selected strains in broth. Isolates of both Staphylococcus aureus and coagulase-negative staphylococci appeared to be potentially susceptible to RP 59500, independent of susceptibility to methicillin or MLS resistance. (S. aureus: methicillin-sensitive, MIC90, 1.0 mg/L; methicillin-resistant, MIC90 1.0 mg/L; coagulase-negative staphylococci: methicillin-sensitive, MIC90 0.5 mg/L). Lancefield group A, B, C and G streptococci (MIC50 0.5 and MIC90 1.0 mg/L) and Streptococcus pneumoniae (MIC50 0.5 and MIC90 1.0 mg/L) appeared to be susceptible to RP 59500. Some Streptococcus spp. and enterococci as well as Listeria monocytogenes were inhibited by a higher concentration of RP 59500 (enterococci: MIC90 4 mg/L, range 0.125-16 mg/L). Comparatively low MICs were seen when Legionella spp., Neisseria gonorrhoeae and Gardnerella vaginalis were tested. Broth dilution MIC/MBC determinations showed no evidence of tolerance, as MIC values were within two dilutions of MBC values. RP 59500 might be a useful compound in the treatment of infections caused by a range of Gram-negative and Gram-positive bacteria, including those resistant to methicillin and/or macrolides.

Coagulase

Activity of RP 59500, a new parenteral semisynthetic streptogramin, against staphylococci with various mechanisms of resistance to macrolide-lincosamide-streptogramin antibiotics.

RP 59500 is a semisynthetic streptogramin (Sg) composed of two synergic components: RP 57669 and RP 54476. The activities of RP 59500, RP 57669 and RP 54476 were tested against 20 strains of staphylococci susceptible to macrolide, lincosamide and streptogramin antibiotics (MLS) and against strains exhibiting different MLS resistance mechanisms. RP 59500 was active against 14 strains harbouring emrA or ermC genes which were inducibly or constitutively resistant to erythromycin (MICs of 0.5-2 mg/L). Neither RP 59500, RP 57669 nor RP 54476 induced MLSB resistance. Constitutive mutants appeared at frequencies of 10(-7)-10(-8) when two MLSB-inducible strains of staphylococci were exposed to 40 mg/L each of clindamycin and RP 57669. No such mutants appeared on plates containing RP 59500 or RP 54476. The emergence of mutants was prevented if the cultures were exposed to RP 54476 (40 mg/L), indicating that such mutants are unlikely to be selected in vivo by RP 59500. However, for some constitutive mutants, MBCs of RP 59500 were as high as 8 mg/L. Strains producing acetyltransferase and hydrolase, inactivating SgA- and SgB-type antibiotics respectively, were resistant to RP 59500, RP 57669 and RP 54476. Production of Pincosamide nucleotidyltransferase-4, which inactivates lincosamides, had no effect on the MICs of RP 59500, RP 57669 and RP 54476.

Anti-Bacterial Agents

Vancomycin resistance gene vanC is specific to Enterococcus gallinarum.

Nearly all strains of Enterococcus gallinarum are resistant to low levels of vancomycin. The glycopeptide resistance gene vanC from E. gallinarum BM4174 has recently been cloned and sequenced. A probe specific for vanC hybridized with a 2.7-kb EcoRI and a 4.5-kb HindIII fragment of total DNA from the 42 strains of E. gallinarum studied. No homology was detected with DNA of strains belonging to other species intrinsically resistant to vancomycin, including Enterococcus casseliflavus, a species that expresses a vancomycin resistance phenotype similar to that of E. gallinarum. No hybridization with DNA of enterococcal strains with acquired resistance to high or low levels of vancomycin was observed. The specificity of the vanC probe allowed us to distinguish E. gallinarum from 12 other species of enterococci, indicating that this probe is a useful tool for species identification within the genus Enterococcus.

Culture Media

Multiple forms of affinity-labeled estrogen receptors in rat distinct pituitary cells.

The presence of multiple monomeric forms has been described for estrogen receptor (ER) in different target tissues. Using [3H]tamoxifen aziridine ([3H]TA) to covalently label ER and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to analyze labeled products, ER forms were investigated in pituitary cytosol and purified nuclei from male rats. ER forms were also compared in cellular extracts from gonadotrope-enriched populations (GP), prepared using the fast method of centrifugal elutriation, and from lactotrope-somatotrope fractions (LSP), obtained by sequential use of both elutriation and Percoll gradient sedimentation. A major labeled protein of 60,000-65,000 mol wt (M(r)) and a minor species of 50,000-55,000 M(r) were found in the pituitary cytosol and nuclear extracts covalently labeled with [3H]TA. The same results were obtained after ER covalent labeling from cellular extracts or intact dispersed cells. In gonadotrope-enriched cell population (greater than or equal to 50% LH-immunoreactive cells), the 65,000 M(r) species is the single unique ER form; in the LSP (80% PRL- and GH-immunoreactive cells), the major TA-labeled species is the 50,000 M(r) form, while the 65,000 M(r) ER is hardly detectable. Thus, the prevalence of 65,000 M(r) protein in the initial cell population can be explained by the higher number of binding sites per gonadotrope than per lactotrope cell. In conclusion, ER heterogeneity is demonstrated in pituitary cell populations. The source of this heterogeneity could be due to 1) different ER mRNAs according to cell type, or 2) a specific posttranslational processing, such as proteolytic activity within lactotrope cells.

Animals

[Small-scale evaluation of the efficacy of growth-regulating insecticides on larvae of the Simulium damnosum complex (Diptera: Simuliidae)].

The efficacy of insect growth regulators was assessed in small scale tests on larvae of the Simulium damnosum complex (Diptera: Simuliidae) in the Ivory Coast. Three compounds [OMS 2015 (triflumuron), OMS 3009 (teflubenzuron), OMS 3013 (chlorfluazuron)] belong to the group of benzoylphenyl-urea substitutes; these IGR's are supposed to inhibit chitin synthesis. Two other compounds are Juvenile Hormone Analogs (JHA's) (OMS 3007 and OMS 3019). The last compound (OMS 3010) is a phenoxycarbamate. The first three compounds had a low efficacy on blackfly larvae, which is consistent with the literature data for another compound of this group: diflubenzuron. The other three compounds (OMS 3007, OMS 3010 and OMS 3019) were much more efficient, OMS 3010 and OMS 3019 showing high activity at low concentrations. These results would justify further studies on the effect of larval age and exposure parameters, and eventually full scale river tests.

Animals

[Prevalence of hepatitis C virus antibodies (tests ELISA 2 and RIBA 2) in a population of pregnant women in France].

We studied the prevalence of anti-HCV antibodies in a population of 2,367 pregnant women attending three public Parisian suburban hospitals. Of this group, 1,614 (68 percent) were French and 753 (32 percent) were immigrant women. The geographic origin of the immigrant population was North Africa (40 percent), West Africa (33 percent), Asia (14 percent), and South Europe (13 percent). Anti-HCV antibodies were tested by the Ortho ELISA second generation test. If present, the Ortho's four-antigen RIBA test and serum alanine aminotransferase determinations were done routinely. The overall prevalence was 1.73 percent. It was 1.55 percent in French women and 2.13 percent in immigrant women. Risk factors associated with anti-HCV were found in 68 percent of the anti-HCV positive French women and in 44 percent of the positive immigrant women. Risk factors were significantly more frequent in anti-HCV positive women in both groups. Among the 41 women with a positive ELISA test, 16/25 French women (64 percent) and 8/16 immigrants (50 percent) had a positive four-antigen RIBA test. Thus, the prevalence of a positive RIBA test was similar in both groups (0.99 and 1.06 percent), due to a higher proportion of false positive ELISA tests observed in the immigrants. These results show that, in French pregnant women: a) the prevalence of anti-HCV is twice as high as that found in blood donors and is a better estimation of the actual prevalence of these antibodies in France; b) positive ELISA 2 tests are most often confirmed by a positive RIBA 2 test.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent

Effects of combinations of beta-lactams, daptomycin, gentamicin, and glycopeptides against glycopeptide-resistant enterococci.

Activities of combinations of beta-lactams, daptomycin, gentamicin, teicoplanin, and vancomycin against 11 clinical isolates of Enterococcus faecium highly resistant to glycopeptides, three plasmid-cured derivatives, eight E. faecalis and E. faecium transconjugants, and two susceptible recipient strains were tested. A marked synergy between penicillins or imipenem and glycopeptides against the glycopeptide-resistant strains but not against the glycopeptide-susceptible strains was observed by the double-disk agar diffusion assay. The synergy of combinations of amoxicillin, imipenem, penicillin G, or piperacillin with vancomycin or teicoplanin against resistant strains was confirmed by the checkerboard technique. The fractional inhibitory concentration indexes were generally below 0.25, except for one strain of E. faecium resistant to high levels of penicillin G. However, the combinations were not bactericidal as tested by time-killing experiments, and high concentrations (64 micrograms/ml) of amoxicillin, penicillin G, or piperacillin combined with 8 micrograms of vancomycin or teicoplanin per ml tended to be antagonistic. Addition of 4 micrograms of gentamicin per ml to these combinations enhanced their bactericidal effect, but they occasionally remained slightly less effective than beta-lactams associated with gentamicin. The combination of 10 micrograms of daptomycin per ml with gentamicin was bactericidal after 6 h against 11 glycopeptide-resistant strains.

Anti-Bacterial Agents

Influence of low-level resistance to vancomycin on efficacy of teicoplanin and vancomycin for treatment of experimental endocarditis due to Enterococcus faecium.

Emergence of vancomycin-resistant strains among enterococci raises a new clinical challenge. Rabbits with aortic endocarditis were infected with Enterococcus faecium BM4172, a clinical strain resistant to low levels of vancomycin (MIC, 16 micrograms/ml) and susceptible to teicoplanin (MIC, 1 micrograms/ml), and against its susceptible variant E. faecium BM4172S obtained in vitro by insertional mutagenesis (MICs, 2 and 0.5 micrograms/ml, respectively). Control animals retained 8 to 10.5 log10 CFU/g of vegetation. We evaluated in this model the efficacy of vancomycin (30 mg/kg of body weight; mean peak and trough serum levels, 27 and 5 micrograms/ml, respectively), teicoplanin (standard dose, 10 mg/kg; mean peak and trough levels, 23 and 9 micrograms/ml, respectively; and high dose, 20 mg/kg; mean peak and trough levels, 63 and 25 micrograms/ml, respectively), gentamicin (6 mg/kg; mean peak and trough levels, 8.6 and less than 0.1 micrograms/ml, respectively), alone or in combination, given every 12 h intramuscularly for 5 days. Teicoplanin standard dose was as active as vancomycin against both strains. Vancomycin was not effective against E. faecium BM4172 but was highly effective against E. faecium BM4172S (7.5 +/- 1.1 log10 CFU/g of vegetation versus 4.9 +/- 1.0 log10 CFU/g of vegetation for vancomycin against E. faecium BM4172 and E. faecium BM4172S, respectively; P = 0.0012). A high dose of teicoplanin was more effective than vancomycin against E. faecium BM4172 (4.4 +/- 1.8 log10 CFU/g of vegetation versus 7.5 +/- 1.1 log10 CFU/g of vegetation for teicoplanin high dose and vancomycin, respectively; P less than 0.05). Against E. faecium BM4172 glycopeptide-gentamicin combinations were the most effective regimens in vitro and in vivo (2.8 +/- 0.7 and 3.5 +/- 1.3 log10 CFU/g of vegetation for vancomycin plus gentamicin and teicoplanin standard dose plus gentamicin, respectively; P < 0.05 versus single-drug regimens). We concluded that high-dose teicoplanin or the combination of a glycopeptide antibiotic plus gentamicin was effective against experimental infection due to E. faecium with low-level resistance to vancomycin.

Animals

Estradiol negatively regulates secretogranin II and chromogranin A messenger ribonucleic acid levels in the female rat pituitary but not in the adrenal.

Estradiol (E2) effects on the pituitary and adrenal secretogranin II (SgII) and chromogranin A (CgA) proteins and mRNA levels were analyzed in the adult female rat. Animals were ovariectomized or sham-operated for 2 weeks and then daily injected with various doses of 17 beta-E2 (from 5-100 micrograms) for the following week. SgII and CgA levels were determined by Western blot analysis using two specific antisera. Messenger RNA (mRNA) levels were measured by RNA slot blot analysis using specific cDNA probes. Simultaneously, pituitary LH content and gonadotropin subunit (LH beta, FSH beta, alpha) mRNAs were quantified. Ovariectomy promoted a significant increase in pituitary SgII and CgA proteins (2-fold vs. sham-operated animals, P less than 0.01) and a concomitant rise in their mRNA levels (2.5-fold and 4.5-fold for SgII mRNA and CgA mRNA, respectively, P less than 0.01). In the same animals LH beta, FSH beta, and alpha-subunit mRNA levels increased by 20-, 12-, and 6-fold, respectively. Estrogen replacement resulted in a parallel decrease of CgA and LH beta mRNA to the control values, starting from the lowest steroid dose (5 micrograms). The SgII mRNA decrease was initiated only with a higher concentration of E2 (10 micrograms), as was that of alpha-subunit mRNA; yet, the SgII mRNA level remained significantly higher than in the control pituitary, even with the highest steroid dose (P less than 0.05) at variance with the alpha-mRNA level. Concerning protein concentration, the postcastration increase in CgA was fully reverted with 10 micrograms E2 while that of SgII remained unaffected, as was the pituitary LH content. In the adrenal gland, neither the ovariectomy nor the E2 therapy altered significantly the SgII or CgA protein and mRNA concentrations. We conclude that, in rats, 1) ovarian factors regulate the pituitary SgII and CgA protein and mRNA steady-state levels while such factors are inefficient in the adrenal gland, 2) CgA and LH beta mRNAs exhibit the same sensitivity to E2 while SgII and alpha-subunit mRNAs appear less sensitive, and 3) SgII and LH pituitary contents present a similar pattern of variations when the estrogenic status of the animal is modified.

Adrenal Glands