Localization of the nucleotide excision repair gene ERCC6 to human chromosome 10q11-q21.

We have cloned the human DNA excision repair gene ERCC6 by virtue of its ability to correct the uv sensitivity of Chinese hamster overy cell mutant UV61. This mutant is a member of complementation group 6 of the nucleotide excision repair-deficient rodent mutants. By means of in situ hybridization and Southern blot analysis of mouse x human somatic cell hybrids, the gene was localized to human chromosome 10q11-q21. An RFLP detected within the ERCC6 locus can be helpful in linkage analysis.     

Prospective theoretical and experimental study towards electrochemically grafted poly(N-vinyl-2-pyrrolidone) films on metallic surfaces

Quantum chemistry calculations of equilibrium geometry, atomic charges, dipole moment, and frontier orbital energies are carried out on model N-vinyl-2-pyrrolidone, its radical anion and cation, and three protonated derivatives. Attempts to obtain poly(N-vinyl-2-pyrrolidone) films grafted on nickel and platinum electrodes, respectively by cathodic and anodic polarizations, are reported. A thin, covering and insulating film is obtained on the Pt anode, as indicated by surface characterizations carried out with UPS and IRRAS spectroscopies. The fact that the film remains adherent to the metallic surface in spite of a permanent contact with an acetonitrile solution in which poly(N-vinyl-2-pyrrolidone) is soluble suggests that chemisorption has indeed been achieved.     

Biotransformation of ellipticine into 5-formyl ellipticine by Choisya ternata strains

Sixteen Choisya ternata strains of same genotypic origin were examined for their capacities to biotransform ellipticine. Four strains were able to convert the drug into 5-formylellipticine with good yields. The reaction occurred only in agar medium and was different from ellipticine bioconversion by microorganisms or mammals.     

Les profils de composition en oligoéléments des pollens,marqueurs potentiels en cladistique végétale

In botany tentative attempts have been made to classify species, based not on the plant as a whole, but on a particular character, for example pollen characteristics. Pollen, which is involved in the transmission of male genetic information during sexual reproduction, is of particular interest in distinguishing plants and their relationships.

These characters, observed in photon- and scanning electron- microscopy are symmetry, form, number and position of germinating areas. Being genetically stable for a given species, they identify a plant.

Recently, a novel character has been added: the concentration of elements in the exine, determined by electron probe micro-X-ray analysis. It is interesting to follow relationship of plant species, based on the mineral composition of their pollen as a whole, taking into account both plastic and oligoelements, obtained via an inductive coupled plasma spectrometer. Samples of pollen from different plant species, at distinct taxonomic levels, were analyzed; the results were submitted to a normalisation procedure and the standard patterns obtained were investigated by means of a number of numerical techniques (clustering and factor analysis). The aim was to establish if elements could be considered as chemical markers in plant cladistics. The mineral composition in a given species appeared quite stable and this furnished a convenient identification-critorion.

This approach further allows a chemo-taxonomy of species, in as much as organization of families, and perhaps orders, would conform to the phylogenetic proximity-criterion. Hence, we found clusters including pollen of Gramineae, Coniferae, Betulaceae…

This methodology, that should be completed and corroborated on a wider range of samples would provide an original responsee to the problems of identification and classification encountered in a pollen bank. It follows that:

Hence, the working hypothesis that the chemical markers and, in particular, oligo-elements, complete pollen typologies based on morphological and structural characters, appears quite reasonable.

In conclusion, this paper brings a complement to the mineral composition of the exine.

En botanique, des tentatives ont été faites pour classer les espèces non plus à partir d'individus pris dans leur intégralité, mais sur la base d'une de leurs particularités, par exemple à partir des caractéristiques offertes par les pollens. Ainsi, le pollen, dont le rôle est de transmettre l'information génétique mâle de la plante au moment de la reproduction sexuée, présente des caractères de premier ordre utilisables pour distinguer les différentes plantes et pour situer leur parenté et leur filiation.

Ces caractères, que l'on observe en microscopies photonique et électronique à balayage sont la symétrie, la forme, le nombre et la position des zones germinatives. Pour une espèce génétiquement stable, ils constituent la carte d'identité de la plante.

Il est intéressant de continuer à explorer la voie de la structuration des espèces végétales basée sur la connaissance chimique du pollen en déterminant sa composition en oligo-éléments par l'utilisation d'un spectromètre à plasma à couplage inductif. Ainsi, des échantillons de pollens appartenant à diverses espèces végétales situées à différents niveaux taxinomiques ont été analysés et les résultats normalisés sous forme de profils standard ont été soumis à différentes techniques d'analyse statistique mathématique (classification automatique et analyse factorielle). Le but de cette approche est d'apprécier l'aptitude des éléments minéraux à être des marqueurs chimiques en cladistique végétale.

Les résultats obtenus ont permis de constater la relative stabilité du profil en composition minérale au sein d'une même espèce végétale, ce qui donne accès à un critère d'identification commode.

Cette approche permet d'autre part une chimiotaxinomie des plantes étudiées.

Cette approche méthodologique semble susceptible d'apporter une réponse originale aux problèmes de conservation. Les pollens conservés doivent avoir la même composition minérale que les pollens frais.

— the pattern of pollen composition of oligo-elements, calcium and magnesium is a marker in plant cladistics. Since a typology depending on proximity is found between plant species, families and orders.

— an unknown pollen sample may be located, tentatively, on the base of a described model of pollen relation (shortest connexion network — Prim algorithm — and pollen of Bactris-oil palm and Phoenix dactylifera).     

Antioxidant properties of cystic fibrosis sputum

Oxidative stress is a likely contributor to the pathogenesis of cystic fibrosis (CF) lung disease. However, hydrogen peroxide (H(2)O(2)), a physiological oxidant, is not elevated in CF exhalates. H(2)O(2) may be neutralized by antioxidants in CF airway secretions. The H(2)O(2)-detoxifying capacity of CF airway secretions, obtained via sputum induction, was studied in an in vitro H(2)O(2) cytotoxicity model. 16HBE14o- cells were exposed to H(2)O(2) in culture medium containing either 0 or 10% fetal bovine serum (FBS) or 10% CF sputum supernatant (extracted without use of dithiothreitol). The efficiency of H(2)O(2) neutralization was estimated by measuring intracellular oxidant levels (dihydrorhodamine 123) after 2 h and cell viability (propidium iodide) after 24 h of H(2)O(2) exposure. Furthermore, the presence of reduced thiols (DTNB assay) and reduced glutathione (recycling assay) in CF sputum samples was evaluated. CF sputum extracts completely prevented intracellular oxidant accumulation seen in cells incubated with H(2)O(2) in both control media (i.e., 0 or 10% FBS). Furthermore, CF sputum abolished cell death in 16HBE14o- cells exposed to up to 1 mM H(2)O(2). In contrast, there was 100% cytotoxicity in cells exposed to 600 microM H(2)O(2) in both control media. The H(2)O(2)-detoxifying potential of CF sputum was sustained after catalase and heme peroxidases were inactivated by sodium azide, which does not affect glutathione peroxidase. In addition, reduced protein thiols were found in abundance in CF sputum. In conclusion, CF sputum is capable to neutralize H(2)O(2) and abundant reduced thiols and/or glutathione peroxidase are fully sufficient to detoxify H(2)O(2).     

Spectrum of CFTR mutations on Réunion Island: impact on neonatal screening

The large heterogeneity in the cystic fibrosis (CF) gene is the main difficulty for genotype characterization. Numerous studies have reported considerable variations in frequencies of CF transmembrane conductance regulator (CFTR) mutations in different populations, such as African, Asian, or European populations. To completely characterize the spectrum of mutations in the CFTR gene in the Réunion Island population, we screened 228 CF chromosomes using denaturing high-pressure liquid chromatography and denaturing gradient gel electrophoresis following by direct sequencing. We identified 27 mutations, accounting for 93% of CF chromosomes. They included three novel mutations (M1T, 3121-3C-->G, and L1324P), which are described in this paper. The detection of such a high proportion of Réunion Island CFTR mutations is important for improving neonatal screening of CF on Réunion Island.     

Vitamin E prevents cell death induced by mild oxidative stress in chicken skeletal muscle cells

Apoptosis and necrosis are two forms of cell death that can occur in response to various agents and oxidative damage. In addition to necrosis, apoptosis contributes to muscle fiber loss in various muscular dystrophies as well participates in the exudative diathesis in chicken, pathology caused by dietary deficiency of vitamin E and selenium, which affects muscle tissue. We have used chicken skeletal muscle cells and bovine fibroblasts to study molecular events involved in the cell death induced by oxidative stress and apoptotic agents. The effect of vitamin E on cell death induced by oxidants was also investigated. Treatment of cells with anti-Fas antibody (50 to 400 ng/mL), staurosporine (0.1 to 100 microM) and TNF-alpha (10 and 50 ng/mL) resulted in a little loss of Trypan blue exclusion ability. Those stimuli conducted cells to apoptosis detected by an enhancement in caspase activity upon fluorogenic substrates but this activity was not fully blocked by the caspase inhibitor Z-VAD-fmk. Oxidative stress induced by menadione (10, 100 and 250 muM) promoted a significant reduction in cell viability (10%, 20% and 35% for fibroblasts; 20%, 30% and 75% for muscle cells, respectively) and caused an increase in caspase activity and DNA fragmentation. H2O2 also promoted apoptosis verified by caspase activation and DNA fragmentation, but in higher doses induced necrosis. Vitamin E protected cells from death induced by low doses of oxidants. Although it was ineffective in reducing caspase activity in fibroblasts, this vitamin diminished the enzyme activity in muscle cells. These data suggested that oxidative stress could activate apoptotic mechanisms; however the mode of cell death will depend on the intensity and duration of the stimulus, and on the antioxidant status of the cells.