Cloning and sequencing of mRNAs coding for two adult alpha globin chains of the salamander Pleurodeles waltlii

A cDNA library of erythrocyte mRNAs was established from immature red blood cells of the adult amphibian, Pleurodeles waltlii (urodele; salamander). The cDNA clones corresponding to the four adult globin chains were first identified and characterized by positive selection and the cDNAs derived from the two (major and minor) alpha-globin chains sequenced. The sequences presented contain both the complete 3'-noncoding region and the coding region of both chains, with the exception of the first nine codons of the minor alpha-chain, and a portion of the 5'-noncoding region of the major chain. The amino acid sequences of the encoded alpha-globin polypeptides have been deduced and compared with those of Xenopus laevis and of man. These comparative studies suggest that the alpha-globins of Pleurodeles waltlii and Xenopus laevis may have diverged from a common ancestral gene at the time when mammalian and amphibian lines diverged, and that they then evolved separately. Duplication of the alpha-gene, which is responsible for the polypeptide heterogeneity, appears to have occurred earlier in Pleurodeles waltlii than in Xenopus laevis.     

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.

     

Synthesis of hydroxy fatty acids from 4, 7, 10, 13, 16, 19-[1-14C] docosahexaenoic acid by human platelets

Human platelets incubated in the presence of 54 microM [1-14C]22:6 produced hydroxydocosahexaenoic acid (HDHE) at about half the rate with which 12-hydroxy-5,8,10,14-eicosatetraenoic acid is produced from [1-14C]arachidonic acid. More than 90% of the radioactivity in HDHE was distributed among two major isomers, 14-HDHE and 11-HDHE. The production of HDHEs was unaffected by indomethacin but completely inhibited by 5,8,11,14-heneicosatetraynoic acid, which suggests that the hydroxy fatty acids are produced by lipoxygenase. The proportions of HDHE isomers varied with the concentration of 22:6. The ratio 14-HDHE/11-HDHE was higher at 6.8 microM 22:6 than when platelets were incubated with 54 microM 22:6. It is suggested that the amounts of these isomers produced will depend both on the availability of 22:6 as well as by competition of this acid with other acids for lipoxygenase.     

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.     

Chromosomal location of the active NORs in theSteropleurus martorelli complex

The chromosomal location of the active NORs has been analyzed by a silver impregnation procedure in theSteropleurus martorelli complex. A primary NOR, which is always present at the first meiotic prophase, has been found in each of the four described races. In addition to this, all races possess one or two secondary NORs which are less active than the former and can be occasionally shown. Usually only one of the two homologous chromosomes has been found to be involved with nucleolus organisation.These results are discussed in relation to hypotheses on the chromosome differentiation of this species complex.     

Salt-dependent conformational transitions in the self-complementary deoxydodecanucleotide d(CGCAATTCGCG): Evidence for hairpin formation

Differential scanning calorimetry (DSC), temperature-dependent uv-absorption spectroscopy, and temperature-dependent CD were used to monitor and characterize the salt-dependent, thermally induced structural transitions in the deoxydodecanucleotide d(CGCGAATTCGCG). At the high oligomer concentrations required for DSC, the calorimetric scans revealed a single, monophasic transition curve at all salt concentrations. Based on previous nmr melting studies under similar conditions, we conclude that these monophasic transitions correspond to the cooperative duplex-to-single-strand conversion of the dodecamer. By contrast, at the lower oligomer concentrations used for the spectroscopic studies, the shapes of the uv and CD melting curves were found to depend on the concentration of the added salt. At high salt (≥0.1M Na⁺), a single, monophasic transition curve was observed. At lower salt (?0.01M Na⁺), the CD and uv melting curves exhibit biphasic behavior. Based on the concentration dependence, the enthalpy, and the cooperativity of each transition in the biphasic curve, we conclude that at low salt and low oligomer concentrations, the dodecamer melts in a sequential manner involving initial disruption of a duplex structure and subsequent disruption of a hairpin structure.