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Biomedical subjects

C Rouquette

Publications and source records attributed to C Rouquette.

At least 19 recordsLinked to original sources

The pathogenesis of infection by Listeria monocytogenes.

Listeria monocytogenes is a Gram-positive bacterium responsible for severe infections in human and a large variety of animal species. It is a facultative intracellular pathogen which invades macrophages and most tissue cells of infected hosts where it can proliferate. The molecular basis of this intracellular parasitism has been to a large extent elucidated. The virulence factors, including internalin, listeriolysin O, phospholipases and a bacterial surface protein, ActA, are encoded by chromosomal genes organised in operons. Following internalisation into host cells, the bacteria escape from the phagosomal compartment and enter the cytoplasm. They then spread from cell to cell by a process involving actin polymerisation. In infected hosts, the bacteria cross the intestinal wall at Peyer's patches to invade the mesenteric lymph nodes and the blood. The main target organ is the liver, where the bacteria multiply inside hepatocytes. Early recruitment of polymorphonuclear cells lead to hepatocyte lysis, and thereby bacterial release. This causes prolonged septicaemia, particularly in immunocompromised hosts, thus exposing the placenta and brain to infection. The prognosis of listeriosis depends on the severity of meningoencephalitis, due to the elective location of foci of infection in the brain stem (rhombencephalitis). Despite bactericidal antibiotic therapy, the overall mortality is still high (25 to 30%).

Actins

An iron-dependent mutant of Listeria monocytogenes of attenuated virulence.

A bank of Tn917-insertional mutants from the facultative intracellular pathogen Listeria monocytogenes was screened by an original method based on bacterial growth on synthetic medium under iron-limiting conditions. One mutant, whose in vitro growth in synthetic medium was specifically dependent upon the availability of iron in its environment, was isolated and characterized. The insertional event occurred in a non-coding region, upstream of a rrn operon and located within a 1100-kb NotI fragment of the physical map, where the virulence genes already identified in L. monocytogenes were also present. Protein analysis by SDS-PAGE revealed a pleiotropic effect of the insertional event on cell-associated proteins, suggesting a polar effect of the transposon on adjacent unknown gene(s). The virulence in the mouse of this mutant was strongly impaired, although it was capable in vitro of growing intracellularly and of spreading from cell to cell, as shown by the production of lytic plaques on cell culture.

Animals

[A rapid method for the determination of cyclosporin A in blood by high performance liquid chromatography].

The purpose of this paper is to propose a method for the determination of cyclosporin in whole blood by means of HPLC with UV detection. After a one step extraction into ether, the organic layer is washed NaOH and evaporated to dryness. The residue is reconstituted in a mixture of acetonitrile and water (40/60) (V/V) and injected into the chromatograph. The C8 reversed phase column is warming at 75 degrees C temperature. The mobile phase consists of acetonitrile and water (60/40) (V/V) and the wavelength is set at 210 nm. The technique is rapid, sensitive, reproducible and well suited for therapeutic monitoring and for pharmacokinetic studies.

Chromatography, High Pressure Liquid

[Plasma assay of haloperidol by high performance liquid chromatography].

Haloperidol is extracted with hexane-isoamyl alcohol in the presence of two internal standards, chlorohaloperidol and spiroperidol. For chromatographic separation, a C18 reversed-phase column and a mobile phase of acetonitrile/methanol/phosphate buffer are used. Detection is by UV absorption spectrophotometry at 245 nm, and the limit of detection was set to 1 ng/ml. To study hydroxyhaloperidol, a metabolite of haloperidol which has much less UV-absorption than haloperidol, detection must be seat 220 nm.

Chromatography, High Pressure Liquid