Contribution à l'étude de la structure de la chromatine: I - Etude physico-chimique de la stabilité de la chromatine

Physicochemical studies of calf thymus chromatin were performed on micromicellar suspensions by thermal denaturation. These diluted suspensions were obtained, by a controlled shearing method, from a compact gel chromatin. Sedimentation and free-flow electrophoresis determined the size distribution of these particles. The most important result is a new transition on the melting profiles corresponding to a sudden increase of solution turbidity. This chromatin solution transition occurs at a higher temperature than usual DNA transition. The degree of « turbidity transitiondiminishes with micelle size but disappears when they are very mildly degraded by DNAases and when F₁ histone fraction is removed.This transition is not only size dependent but also depends on the micellar structure. This phenomenon is interpreted as an excluded volume effect by contact between compact and native regions of nucleoprotein micelles and denatured coils of DNA. Our study tried to show that the degree of turbidity transition can be a criterion of chromatin native structure.     

Etude de quelques propriétés de la phosphofructokinase érythrocytaire de l'homme normal

Human erythrocyte phosphofructokinase was purified 150 fold by DEAE cellulose adsorption and ammonium sulfate precipitation.At pH 7,5 the enzyme exhibits allosteric kinetics with respect to ATP, fructose 6 phosphate, and Mg²⁺.ATP at high concentration acted as an inhibitor and ADP, 5′AMP, 3′,5′, AMP, acted as activators. Both effectors seemed to decrease the homotropic interactions beetween the fructose 6 phosphate molecules.The activators increased the affinity of phosphofructokinase for the substrate (F6P), the inhibitor decreased it.These ligands had no effect on the maximum velocity of the reaction except in the case of ADP.Interactions between the substrates and the effector ligands on the enzyme were considered in terms of the Monod - Changeux - Wyman model for allosteric proteins.With GTP and ITP, no inhibition was observed. At saturing concentration of GTP, ATP still inhibited phosphofructokinase.Both 3′5′ AMP and fructose 6 phosphate increased the concentration of ATP required to produce an inhibition of 50 %.Citrate, like ATP, inhibited phosphofructokinase by binding most likely at the same allosteric site. Erythrocyte phosphofructokinase is inhibited by 2–3 DPG.The study of the relation log V max = f (pH) suggested, that the active center contains at least one imidazole and one sulfhydryl group.     

Sur la stéréospécificité de la ribonucléoside-diphosphate-réductase: préparation de désoxyribonucléosides pyrimidiques deutérés

Methyl 2-deoxy-3,5,6-tri-O-p-nitrobenzoyl- -ribo-hexofuranoside was converted into the glycosyl halide which was then condensed with 2,4-bis(trimethylsilyloxy)-pyrimidine in the presence of mercuric chloride to give, after alkaline methanolysis, 1-(2-deoxy-β- -ribo-hexofuranosyl)uracil, in a yield too low for the reaction to be applied to deuterated compounds. Methyl 2-deoxy- -allofuranoside-2-d was degraded into methyl 2-deoxy- -arabinofuranoside-2-d. Its di-p-nitrobenzoate was converted into the glycosyl halide which was coupled with 2,4-bis(trimethylsilyloxy)-pyrimidine to give, after alkaline hydrolysis, deuterated deoxyuridine. Thiationammonolysis of a mixture of the 3′,5′-di-p-nitrobenzoates of the latter compound and its anomer gave the corresponding deoxycytidines. Comparison of the n.m.r. spectra of these compounds to that of deuterated deoxycytidine, prepared by the enzymic reduction of cytidine 5′-pyrophosphate with the Escherichia coli system in the presence of deuterium oxide confirmed that the substitution of a hydroxyl group by a hydrogen atom proceeds with retention of configuration.     

Le système protéolytique de Penicillium roqueforti: II - Purification et propriétés de la protéase acide

An extracellular protease has been isolated from the culture medium of Penicillium roqueforti. The enzyme was purified by precipitation with ammonium sulfate, filtration on Bio-gel P 100 columns and chromatography on D.E.A.E.-cellulose columns The purified preparation was homogenous by gel filtration on Bio-gel P 60 and electrophoretical analysis at pH 9.0 and 5.0.The protease exhibited the properties of an acid protease: the optimum pH was 3.5 for casein or hemoglobin hydrolysis and for bovine trypsinogen activation; at 40°C, the enzyme is most stable in the range of pH 3.5 to 5.5; the optimum temperatures was 50°C.E.D.T.A., D.F.P. and sulfhydryl reagents induced no inhibition.The enzyme exhibited a milk clotting activity that was fifty times weaker that the activity of rennin. Its molecular weight, estimated by gel filtration was 33 400. Amino acid composition is: Lys₁₅, His₂, Arg₁, Trp₅, Asp₃₃, Thr₂₇, Glu₁₆, Pro₁₀, Gly₄₀, Ala₂₅, Cys₂, Val₂₁, Ile₂₀, Leu₂₁, Tyr₁₄, Phe₁₉.The properties of this protease were closely similar to that of P. janthinellum and Aspergillus oryzae.     

Etude de la fertilité des eaux marines au moyen de tests biologiques effectués avec des cultures d'algues. II. Limitation nutritionnelle et viabilité de l'inoculum

Ten cultures of phytoplankters, including four strains of Skeletonema costatum from different origins, were used to improve some aspects of the bioassay technology. Special attention was paid to the preliminary nutrient limitation of the inocula. When the cells are maintained in nutrient starvation, their carbon, nitrogen, carbohydrates, proteins and above all chlorophyll a contents decrease. The minimum appears after a range of 2–7 days, according to species. The survival of these limited cells and their capacity to give rise to active growing cultures when sub-cultured are different with species, but efficiency in the inocula they provide usually can occur only until the minimum content in cellular components appears. Starved cells and enriched cultures of Chaetoceros lauderi and Skeletonema costatum were used to inoculate several samples of sea water, in order to test the effect of the starvation on the experimental results. It appears that the starved cells increase the sensitivity of the method, but they are more susceptible to substances limiting their growth. In the opinion of the authors the best way would be to use both starved and enriched cells as inocula, but, when this is impossible, cautiously starved cells should be used with unpolluted sea waters.     

Applications biologiques de la spectroscopie Raman de résonance et de la spectroscopie micro-Raman de résonance

Advantages are given to illustrate the possibilities of resonance Raman spectroscopy in the studies of biological systems.A relation giving the size of porphinato-core of metalloporphyrins from resonance Raman frequency is proposed. It can be applied to heme, even in lived state.By illuminating the sample through a microscope, resonance Raman effect can be collected from 1 (μm)² of sample. Spectra of a vegetal-cell wall (pimento) and of a red corpuscle (hemoglobin) are shown as examples.     

Activité oxydasique de l''oxydoréductase de la xanthine, spécifique de la classe des mammifères

In every species other than mammals xanthine oxidoreductase behaves as a dehydrogenase, never as an oxidase. In three mammalian species, the enzyme acts intracellularly as a dehydrogenase, but its class-specific ambivalence allows its extracellular conversion into an oxidase.

Résumé

En dehors de la classe des Mammifères, l'oxydoréductase de la xanthine ne se comporte jamais comme une oxydase. Chez les Mammifères, une ambivalence de la molécule permet l'expression d'une activité déshydrogénasique (NAD) dans la cellule, et d'une activité oxydasique en dehors de la cellule.     

La diversité floristique de la végétation psammophile de la région de Tlemcen (nord–ouest Algérie)

This study is devoted to the analysis of the psammophyte of the coastal and semi-continental dunes in Tlemcen. Interesting results have been obtained, in particular, on the biological and ecological aspects of the psammophyte. The interpretation from Factoriel analysis of correspondences enabled us to identify the different phytosociological classes (Cakiletea maritimae, Ammophiletea, Quercetea ilicis, Therobrachypodietea and Stellarietea mediae). Some of these classes (Cakiletea maritimae and Ammophiletea) inhabit, exceedingly well, the embryonic dunes. Some species (Therobrachypodietea) colonize the quickset dunes. Lastly, some others (Quercetea ilicis) settle in the more mature and stable dunes. By using the phytosociological and phytodynamical data, we have been able to understand the vegetation and its diversity. To cite this article: H. Stambouli-Meziane et al., C. R. Biologies 332 (2009).     

Evolution des activités enzymatiques dans le tractus digestif au cours de la vie larvaire de la sole. Variations des protéinogrammes et des zymogrammes

The activities of digestive enzymes were studied during the course of larval development (from hatching to 30 days) in the sole, Solea solea. No glycosidase activity was present, but alkaline proteolytic activity increased from hatching to day 5, then decreased until day 15. In the acid range, proteolytic activity was relatively stable from the 10th to the 30th day. There were also changes in the number of esterase-like isozymes which increased between day 20 and 30, with minimum activity observed during metamorphosis.     

Déterminisme de la couleur des hôtes parasités par Trichogramma fuentesi

Résumé A Cuba, les oeufs h?tes parasités parTrichogramma fuentesi Torre (Hymenoptera: Trichogrammatidae) deviennent soit noirs, soit marrons. Une série de croisements entre Trichogrammes a permis de montrer que le déterminisme de ce caractère est sous la dépendance d'un locus portant deux allèles: Bh, dominant, confère une couleur noire à l'h?te; bh, récessif, confère une couleur marron. L'allèle bh étant plus fréquent dans la région orientale de Cuba, la plus chaude, nous avons suivi l'évolution des fréquences des deux allèles dans des populations expérimentales polymorphes placées à 23 °C et 30 °C. Dans toutes les conditions testées, bh a été désavantagé par rapport à Bh, sauf à 30 °C lorsque sa fréquence initiale était élevée (90%). La température n'explique donc certainement pas à elle seule le maintien du polymorphisme à Cuba.

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In Cuba, during its larval development inside its host egg,Trichogramma fuentesi turns its host egg either black or brown. Crossings carried out between the two forms ofT. fuentesi allowed to show that this character is determined by one locus with two alleles: Bh, dominant, colours the host black; and bh, recessive, colours the host brown. The bh allele is more frequent in the eastern region of Cuba which is also the warmest, than in the western region. We studied the evolution of frequencies of the two alleles in different experimental polymorphic populations at 23 °C and at 30 °C. In all tested conditions, bh was at a disadvantage by comparison with Bh, except at 30 °C when its initial frequency was high (90%). Thus, temperature alone should certainly not explain the maintenance of the observed polymorphism in Cuba.

     

Sur la taxonomie et l'éthologie de quelques Orbioninés (Isopoda Epicaridea).

Studies on the Taxonomy and the Ethology of some Orbioninae (Isopoda Epicaridea) Epicarid isopods of the sub-family Orbioninae are parasitic on peneids. In this paper, two new species are described, and additional information on ten species is provided.     

Étude polarographique de la mobilisation du fer de la ferritine

Polarographic study of the mobilization of ferritin ironPolarographic study allows to propose a model for mobilization of ferritin iron: an equilibrium exists between iron core and small quantities of iron outside the protein.These iron atoms would be lying on electron acceptor sites including SH groups. The number of sites is dependent on iron content of ferritin.Therefore, the iron could be removed by the action of reducing agents such as xanthine oxidase or ascorbic acid, and then chelated by a complexing agent.