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Biomedical subjects

J Carrière

Publications and source records attributed to J Carrière.

18 recordsLinked to original sources
PubMed1999

Amphotericin B susceptibility testing of Candida lusitaniae isolates by flow cytofluorometry: comparison with the Etest and the NCCLS broth macrodilution method.

A flow cytofluorometric susceptibility test (FCST) was used for rapid determination of the susceptibility of Candida lusitaniae isolates to amphotericin B. The test is based on the decrease in fluorescence intensity of cells stained with 3,3'-dipentyloxacarbocyanine iodide (DiOC5(3)), a membrane potential-sensitive cationic dye, after drug treatment. A total of 58 C. lusitaniae clinical isolates including strains known to be amphotericin B-resistant on the basis of in-vivo and/or in-vitro data were tested. MICs were determined concurrently by the NCCLS broth macrodilution method and the Etest, both with antibiotic medium 3. Regression analysis demonstrated that the data from the FCST and the Etest were better correlated (r = 0.93, n = 59, P < 0.001) than those from the FCST and the NCCLS method (r = 0.63, n = 59, P < 0.001). The FCST readily identified a series of putatively susceptible and resistant isolates. Our study points out the advantages of the flow cytometry approach in antifungal susceptibility testing of yeasts, since speed remains a major problem in conventional tests.

Amphotericin B↗
PubMed1998

Estrogen response elements can mediate agonist activity of anti-estrogens in human endometrial Ishikawa cells.

Anti-estrogens like hydroxytamoxifen (OHT) have mixed agonist/antagonist activities, leading to tissue-specific stimulation of cellular proliferation. Partial agonist activity of OHT can be observed in vitro in endometrial carcinoma cells like Ishikawa. Here, we have compared several anti-estrogens (including extensively characterized OHT and pure anti-estrogens such as ICI164, 384 and RU58,668, which are devoid of uterotrophic activity) for their capacity to stimulate promoters containing estrogen response elements (EREs) or AP1-binding sites (12-O-tetradecanoylphorbol-13-acetate response elements, TREs), the two types of DNA motifs known to mediate transcriptional stimulation by estrogen receptors. Assays were performed in Ishikawa cells either by transient transfection or by using cell lines with stably propagated reporter vectors. In transient transfection experiments, none of the anti-estrogens displayed agonist activity on the promoters tested. In contrast, significant transcriptional stimulation was observed with low concentrations of OHT and RU39,411 in Ishikawa cells stably propagating reporter constructs containing a minimal ERE3-TATA promoter. In addition, micromolar concentrations of OHT, but not of RU39,411, stimulated stably propagated AP1-responsive reporter constructs. No transcriptional stimulation of ERE- or TRE-containing promoters was observed with the pure anti-estrogens ICI164,384 and RU58,668. These results indicate that the presence of estrogen response elements in promoters is sufficient to mediate cell-specific agonism of anti-estrogens at the transcriptional level, and that stimulation of AP1 activity may be restricted to a subset of anti-estrogens possessing agonist activity on EREs. In addition, our results suggest that transient transfections do not fully recapitulate in vivo conditions required to observe agonist activity of anti-estrogens.

Animals↗
PubMed1996

Mutations of non-canonical base-pairs in the 3' major domain of Escherichia coli 16 S ribosomal RNA affect the initiation and elongation of protein synthesis.

Three single-point mutations and two multibase substitutions were introduced into the lower half of the 3' major domain of Escherichia coli 16 S ribosomal RNA. The three single mutations were located in helix 29 (U1341C) or in helix 43 (U1351C or A1357C) and replaced highly conserved non-canonical base-pairs with Watson-Crick base-pairs. The two multibase substitutions were located at the base of helix 42, where they transformed an irregular portion into a Watson-Crick [correction of Waston-Crick] segment. Each of the single mutations could be expressed in vivo from the rrnB operon of a multicopy plasmid under control of constitutive promoters, and none of them affected growth-rate. However, mutation A1357C, but not U1341C or U1351C, severely retarded cell growth, when expressed together with another mutation in the upper half of the 3' major domain, C1192U. The latter mutation is located in helix 34 and abolishes the binding of spectinomycin, a protein synthesis inhibitor. The proportion of mutated ribosomes was high in polysomes, suggesting that the A1357C and C1192U double mutation did not affect the initiation but rather the elongation of protein synthesis. The effect of the double mutation reveals a functional interplay between helices 34 and 43. Furthermore, an interaction between helices 34 and 43 was also suggested by studies of protection by spectinomycin against chemical attack, that showed that the binding site of spectinomycin was restored with ribosomes bearing another double mutation, U1351C and C1192U. In contrast to the single mutations, the multiple mutations in helix 42 could not be expressed in vivo under control of the strong constitutive promoters, but could be expressed under control of the weaker, thermoinducible lambda PL promoter. They did not affect cell growth, whether expressed in the absence or the presence of the C1192U mutation. However, under conditions where protein synthesis depended exclusively on ribosomes with plasmid-encoded rRNA, cells transformed with plasmids altered in helix 42 could not grow. Analysis of the plasmid-borne 16 S rRNA distribution in bacteria transformed with these mutant plasmids showed that mutant 16 S rRNA was present in a high proportion in the free 30 S subunits but was underrepresented in 70 S ribosomes and polysomes. Extension inhibition assays (toeprinting) demonstrated that this altered distribution resulted from an impaired capacity of the mutant 30 S subunits to form translation initiation complexes.

Bacterial Proteins↗
PubMed1995

Dermatoses associated with travel to tropical countries: a prospective study of the diagnosis and management of 269 patients presenting to a tropical disease unit.

The full spectrum of skin diseases related to travel in tropical areas is unknown. We prospectively studied 269 consecutive patients with travel-associated dermatosis who presented to our tropical disease unit in Paris during a 2-year period. The median age of these patients was 30 years; 137 patients were male; 76% of the patients were tourists; 38% had visited sub-Saharan Africa; and 85% had been appropriately vaccinated against tetanus. Cutaneous lesions appeared while the patient was still abroad in 61% of cases and after the patient's return to France in 39%. The diagnosis was definite in 260 cases; 137 of these cases (53%) involved an imported tropical disease. The most common diagnoses were cutaneous larva migrans (25%); pyodermas (18%); pruritic arthropod-reactive dermatitis (10%); myiasis (9%); tungiasis (6%); urticaria (5%); fever and rash (4%); and cutaneous leishmaniasis (3%). Hospitalization was necessary in 27 cases (10%), with a median duration of 5 days (range, 2-21 days). Travelers should be advised on how to avoid exposure to the agents and vectors of infectious dermatoses. Travel first-aid kits should include insect repellents and antibiotics effective against bacterial skin infections.

Adolescent↗
PubMed1991

Contribution of the glutamine 19 side chain to transition-state stabilization in the oxyanion hole of papain.

The existence of an oxyanion hole in cysteine proteases able to stabilize a transition-state complex in a manner analogous to that found with serine proteases has been the object of controversy for many years. In papain, the side chain of Gln19 forms one of the hydrogen-bond donors in the putative oxyanion hole, and its contribution to transition-state stabilization has been evaluated by site-directed mutagenesis. Mutation of Gln19 to Ala caused a decrease in kcat/KM for hydrolysis of CBZ-Phe-Arg-MCA, which is 7700 M-1 s-1 in the mutant enzyme as compared to 464,000 M-1 s-1 in wild-type papain. With a Gln19Ser variant, the activity is even lower, with a kcat/KM value of 760 M-1 s-1. The 60- and 600-fold decreases in kcat/KM correspond to changes in free energy of catalysis of 2.4 and 3.8 kcal/mol for Gln19Ala and Gln19Ser, respectively. In both cases, the decrease in activity is in large part attributable to a decrease in kcat, while KM values are only slightly affected. These results indicate that the oxyanion hole is operational in the papain-catalyzed hydrolysis of CBZ-Phe-Arg-MCA and constitute the first direct evidence of a mechanistic requirement for oxyanion stabilization in the transition state of reactions catalyzed by cysteine proteases. The equilibrium constants Ki for inhibition of the papain mutants by the aldehyde Ac-Phe-Gly-CHO have also been determined. Contrary to the results with the substrate, mutation at position 19 of papain has a very small effect on binding of the inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1973

An appraisal of the bentonite flocculation test in the detection of bovine tuberculosis.

In this further study, an exhaustive investigation into the value of the bentonite flocculation test in detecting tuberculosis in cattle was made on a group of 358 cattle, with known intradermal (ID) tuberculin reactions and with or without tuberculosis lesions. Bentonite sensitized with human old tuberculin (OT) and a purified BCG carbohydrate fraction (BCG-F1) were used. In the group of 126 cows with tuberculosis lesions, 113 were positive to the tuberculin ID test and there were also 113 positive reactions to the flocculation test using OT sensitized bentonite. Only 62 were positive using the BCG-F1 sensitized bentonite. Out of the 232 animals with no visible lesions, 114 reacted positively to the tuberculin ID test, 148 had a titer to the flocculation test using OT sensitized bentonite and 70 were positive when using BCG-F1 sensitized bentonite.

Animals↗