Isolement et sélection de souches d'actinomycètes productrices de substances antifongiques de structure non-polyénique

Résumé Dans le but d'étudier des souches d'actinomycètes productrices de substances antifongiques de structure non-polyénique, 13, échantillons de sol prélevés dans le sud de la France ont été examinés. L'utilisation de milieux sélectifs a permis d'isoler 486 souches d'actinomycètes qui ont été testées vis-à-vis de quatre espèces de champignons et de levures: 18% des souches isolées sont actives sur au moins l'une des espèces utilisées. Parmi celles-ci 14 souches, productrices de substances de structure non-polyénique, ont été sélectionnées après étude des spectres d'absorption en UV des surnageants de culture, des extraits butanoliques de ces surnageants ou des extraits méthanoliques de mycélium. L'utilisation d'un test bactérien de toxicité à court terme (SOS Chromotest) a permis de montrer que 10 souches sur 14 présentent aussi une activité génotoxique.

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Summary In order to study actinomycete strains producing non-polyenic antifungal substances 13 soil samples were collected in S. France. By using selective media 486 strains of actinomycetes were isolated and tested on four species of moulds and yeasts: 18% of the isolated strains were active against one or more of the test organisms. From these isolates 14 producers of non-polyenic antifungal substances were selected by means of u.v. absorption spectra of culture supernatant fractions, butanol extracts of these fractions, or methanol extracts of mycelium. A rapid bacterial toxicity assay (SOS Chromotest) demonstrated that 10 of the 14 selected strains had genotoxic activity.

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Resumen Para el estudio de cepas de actinomicetes productores de sustancias antifúngicas no poliénicas, se recogieron 13 muestras de suelo en el S. de Francia. Utilizando medios selectivos se aislaron 486 cepas de actinomicetes que se ensayaron frente a cuatro especies de mohos y levaduras: 18% de las cepas aisladas mostraron actividad frente a uno o más de los organismos utilizados en el test. De estas cepas se seleccionaron 14 que eran productoras de sustancias antifúngicas no poliénicas mediante el espectro de absorción al U.V. de las fracciones sobrenadantes del cultivo, de extractos butanólicos de dichas fracciones o de extractos metanólicos del micelio. Un ensayo rápido de toxicidad bacteriana (SOS Chromotest) mostró que de las 14 cepas seleccionadas lo tenían actividad genotóxica.

     

Angina pectoris-like pain provoked by intravenous adenosine in healthy volunteers.

In a study to characterise the chest pain induced by adenosine this agent was given as a bolus into a peripheral vein to six healthy volunteers (five men) aged 30-44. On the first day the maximum tolerable dose was determined in each case. On the second day three doses of adenosine (one third, two thirds, and the full maximum tolerable dose) and three doses of saline were given single blind in randomised order. Thereafter aminophylline 5 mg/kg was given and the procedure repeated in a different randomised order. On the third day between two thirds and the full maximum tolerable dose was given followed by 10 mg dipyridamole intravenously and a second injection of the same dose of adenosine. Heart rate and atrioventricular blocks were recorded by electrocardiography. One minute after each dose of adenosine the chest pain was scored. The maximum tolerable dose of adenosine ranged from 10.6 to 37.1 mg. All subjects experienced uneasy central chest pain provoking anxiety. The pain radiated to the shoulders, ulnar aspect of the arms, epigastric area, back, and into the throat. The pain began about 20 seconds after the injection and lasted 10-15 seconds. Increasing the dose of adenosine increased the intensity of the pain. Administration of aminophylline reduced the pain significantly. Second degree heart block was recorded in five of the six subjects during the time that the pain was experienced. After aminophylline no block was observed. Dipyridamole increased the intensity of pain. The duration of second degree heart block increased in four of the subjects, and in two of these third degree heart block occurred. These findings suggest that adenosine released from the myocardium during ischaemia induces angina pectoris by stimulating theophylline sensitive receptors.     

Isolation and NH2-terminal sequence of a highly conserved human and porcine pituitary protein belonging to a new superfamily: Immunocytochemical localization in pars distalis and pars nervosa of the pituitary and in the supraoptic nucleus of the hypothalamus

The isolation and purification of a 21,000-Da (pI 4.9) novel protein from porcine anterior pituitary and whole human pituitary is described. Comparison of the NH₂-terminal sequence of the first 77 and 81 residues of the human and porcine homologs shows only one conservative substitution at residue 12, namely an Ala for a Thr between these two species. Such high sequence homology is also reflected in their amino acid composition. A computer data-bank search using a mutation data matrix and comparison with 338,327 segments of proteins revealed that this substance should be classified as belonging to a new protein superfamily. Immunocytochemical staining, using an antibody produced against a synthetic fragment, revealed the presence of immunostainable material in the anterior and posterior lobe of the pituitary and in the supraoptic nucleus of the hypothalamus. No staining was observed in the intermediate lobe of the pituitary. Furthermore, purified neurointermediate lobe secretory granule preparations were also shown to contain this novel polypeptide.     

Cyclic nucleotide phosphodiesterases in larval brain of wild type and dunce mutant strains of Drosophila melanogaster: isoenzyme pattern and activation by Ca2+/calmodulin

Like adult heads and whole flies, larval brains of wild type Drosophila melanogaster contain two major soluble cyclic nucleotide phosphodiesterases, forms I and II. Larval brains of the learning-defective mutant strain, dunceM11, contain only the form I enzyme. In both wild type and dunce strains the form I enzyme is activated by Ca2+/calmodulin. A time-dependent loss of this Ca2+ activation was observed.     

Characterization of black-pigmented Bacteroides strains isolated from animals

The aims of the study were: the isolation of strains of black-pigmented Bacteroides from the gingival sulcus of different animals, their biochemical and immunological characterization and comparison of their properties for classification within the genus. A total of 104 strains, isolated from cats, dogs, racoons and a jaguar, were characterized on the basis of fermentation of carbohydrates, metabolic end products, haemagglutination studies, enzymatic activities, catalase production and indirect immunofluorescence. No differences were observed between the strains regardless of their animal origin. The strains did not ferment carbohydrates, produce phenylacetic acid, show an array of enzyme activities or agglutinate sheep red blood cells. They were catalase-positive and so differed from the human oral strains of Bact. gingivalis. Immunofluorescence microscopy revealed that the animal strains shared at least one major antigen with Bact. gingivalis but none with Bact. asaccharolyticus. Apart from their catalase activity, the animal strains isolated were similar to those of human Bact. gingivalis strains.     

Maintenance and reversibility of active albumin secretion by adult rat hepatocytes co-cultured with another liver epithelial cell type

When adult rat hepatocytes were co-cultured with another liver epithelial cell type in a medium supplemented or not with fetal calf serum (FCS), it was found that 1. They survived for more than 2 months 2. Albumin secretion levels remained high over the whole culture period 3. Decreased secretion might be reversed 4. This protein secretion activity appeared to be dependent upon both the presence of cell-cell contacts and the production of an extracellular material. The results demonstrate for the first time long-term stabilization and reversibility of a specific function (albumin secretion) at high levels by adult hepatocytes cultured in serum-free medium and suggest that both the presence of other liver cell type(s) and the production of an extracellular matrix are needed for the maintenance of specific functions in cultured hepatocytes.     

Clathrin-associated proteins contain bound nucleotide

An alcohol dehydrogenase isolated from Zymomonas mobilis was found to be activated by ferrous ions but not by zinc, after inactivation with metal-complexing agents. Cobaltous ions also re-activated to a lesser extent. It is suggested that in this species the alcohol dehydrogenase naturally contains iron. Kinetic studies on the iron-treated enzyme indicate an 'alcohol activation' phenomenon, which may have physiological relevance in overcoming product inhibition during fermentation.     

Trypanosoma cruzi: 4-aminopyrazolopyrimidine in the treatment of experimental Chagas' disease

An allopurinol metabolite, 4-aminopyrazolopyrimidine, was tested on two different strains of mice (NMRI-IVIC and C57Bl/6J) that had been infected 4 days earlier with the virulent Ya strain of Trypanosoma cruzi. Low doses of 4-aminopyrazolopyrimidine (0.125-0.500 mg/kg body wt/day) for 10 days induced a significant reduction in parasitemia (direct counts and subinoculation experiments) and increased survival time (without any evidence of toxicity) compared with untreated animals. When tested in vitro, 4-aminopyrazolopyrimidine was sixfold more active than allopurinol as a trypanostatic drug. The low therapeutic doses of 4-aminopyrazolopyrimidine suggest that this drug may be useful in the treatment of acute Chagas' disease.