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2.
Life Sci ; 69(14): 1587-96, 2001 Aug 24.
Article in English | MEDLINE | ID: mdl-11589499

ABSTRACT

Ascorbate and tocopherol are important antioxidants that protect cells against oxidative stress. The interaction of ascorbate and alpha-tocopherol in cells is difficult to detect as both ascorbate and alpha-tocopherol are unstable in vitro in a biological medium. We examined the interactions between human dermal fibroblasts, ascorbate and alpha-tocopherol to determine the effects of the vitamins on growth and cell viability. The interaction of ascorbate and alpha-tocopherol was studied in a fibroblast culture medium during 48h. Ascorbate and alpha-tocopherol were detected by fluorimetry after high-performance liquid chromatography (HPLC). Cell growth and cell viability were studied by cell numeration after trypan blue staining. The ascorbate concentration fell in presence of alpha-tocopherol in cell culture medium under all experimental conditions, with or without cells. Ascorbate partly protected alpha-tocopherol but only in presence of cells. Cell viability was preserved by alpha-tocopherol whereas ascorbate enhanced fibroblast growth. The synergy between ascorbate and alpha-tocopherol corresponds to a consumption of ascorbate which spares alpha-tocopherol but only in presence of cells.


Subject(s)
Ascorbic Acid/pharmacology , Fibroblasts/drug effects , Vitamin E/pharmacology , Ascorbic Acid/analysis , Cell Division/drug effects , Cell Survival/drug effects , Cells, Cultured , Chromatography, High Pressure Liquid , Fibroblasts/cytology , Humans , Spectrometry, Fluorescence , Vitamin E/analysis
3.
J Mol Evol ; 51(4): 374-7, 2000 Oct.
Article in English | MEDLINE | ID: mdl-11040288

ABSTRACT

Wolbachia are symbiotic bacteria altering reproductive characters of numerous arthropods. Their most recent phylogeny and classification are based on sequences of the wsp gene. We sequenced wsp gene from six Wolbachia strains infecting six Trichogramma species that live as egg parasitoids on many insects. This allows us to test the effect of the addition of sequences on the Wolbachia phylogeny and to check the classification of Wolbachia infecting Trichogramma. The six Wolbachia studied are classified in the B supergroup. They confirm the monophyletic structure of the B Wolbachia in Trichogramma but introduce small differences in the Wolbachia classification. Modifications include the definition of a new group, Sem, for Wolbachia of T. semblidis and the merging of the two closely related groups, Sib and Kay. Specific primers were determined and tested for the Sem group.


Subject(s)
Bacterial Outer Membrane Proteins/genetics , Phylogeny , Wolbachia/classification , Wolbachia/genetics , Animals , Asia , DNA Primers , Europe , Polymerase Chain Reaction/methods , South America , Species Specificity , Symbiosis , Wasps/microbiology
4.
J Invertebr Pathol ; 76(1): 20-5, 2000 Jul.
Article in English | MEDLINE | ID: mdl-10963399

ABSTRACT

Fluorescence in situ hybridization was tested to specifically detect symbionts of the genus Wolbachia in Trichogramma and to allow for semiquantitative estimations of symbiont abundance. Extraction solutions used for horizontal transfers of symbionts contain a high abundance of Wolbachia, but Wolbachia have a low and decreasing abundance in microinjected lines (transfected lines). Moreover, eggs of microinjected lines were shown to be polymorphic for the infection. In naturally infected lines, Wolbachia are localized at the posterior pole of the eggs; they are scattered during the early stages of larval development and then concentrated in the ovaries at the end of the female pupal development. Scattering and concentration are probably not active but rather the result of replications or morphogenesis. Conversely, Wolbachia are not concentrated at the posterior pole of eggs in microinjected lines. Comparison of the within-family and between-family variances of the symbiont abundance in a microinjected line did not lead us to conclude that this character shows a genetic variability.


Subject(s)
Wasps/microbiology , Wolbachia/growth & development , Animals , Female , Genetic Variation , Microinjections , Ovary/microbiology , Solutions , Symbiosis , Wasps/growth & development , Wolbachia/genetics , Wolbachia/isolation & purification
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