Characterization of Methanosarcina mazeii TMA isolated from a paddy field soil

We isolated a methanogenic strain, designated as strain TMA (=DSM 9195), from an enrichment culture inoculated with a Japanese paddy field soil. Strain TMA was Gram positive and strictly anaerobic. Cell shape was pseudosarcina-like, and cells were nonmotile. The strain was able to use methylamines, methanol, H₂–CO₂, and acetate as substrates for methanogenesis, but did not utilize formate. The optimum temperature and optimum pH were 30–37°C and 6.5–7.5 respectively. The G+C content of the DNA was 42.1 mol %. Strain TMA had DNA-DNA hybridization values of more than 80% with Methanosarcina mazeii S-6^T (T = type strain). On the basis of phenotypic and genotypic characteristics, we identified strain TMA as M. mazeii. This is the first methylotrophic methanogen isolated from a paddy field soil and identified to the species level.     

Optimization of a medium for the production of 1,2-epoxytetradecane by Nocardia corallina B-276

Summary A medium for the production of 1,2-epoxytetradecane from 1-tetradecene by Nocardia corallina B-276 was optimized. The activity of cells producing 1,2-epoxytetradecane increased when cell growth was suppressed by limiting nitrogen or potassium ion in the medium. Mg²⁺ was found to be essential for the production of 1,2-epoxide. Cell mass was increased, without reducing production of 1,2-epoxide, by increasing the concentration of yeast extract and limiting the concentration of potassium ion. The concentration of 1,2-epoxytetradecane reached 80 g/l in 6 days after optimization and the yield of 1,2-epoxytetradecane was 65 mol% based on consumed 1-tetradecene. Long chain 1,2-epoxyalkanes containing 13–17 carbon atoms also were produced under these conditions.     

Isolation and identification of substances inducing formation of tracheary elements in cultured carrot-root slices

Data are presented on the cytokinin status of seeds and seed components, at different stages of development in Phaseolus coccineus L., as determined with the soybean callus growth bioassay: A change in cytokinin types according to developmental stage occurred: from biologically very active less polar types (zeatin=Z) at early stages to more polar types (zeatin glucoside=Z₉G and zeatin riboside=Zr), with relatively low biological activity, at intermediate and late stages of seed development: When cytokinins were analyzed separately in embryos (embryo proper) and suspensors at two embryonic stages: heart-shaped (A) and middle cotyledonary embryos (stage B) respectively, it was found that: i) at stage A, the suspensor showed cytokinin activity at the level of Z, 2iPA (2-isopentenyladenosine) and Zr, whereas more polar cytokinins (Z₉G, Zr) were present in the embryo; ii) at stage B, when the embryo seems to become autonomous for cytokinin supply, there was a relative abundance of active cytokinins (Z, 2iPA) in the embryo to which Z₉G activity in the suspensor corresponded. It is concluded that the suspensor plays an essential role in embryogenesis by acting as a hormone source to the early embryo.Abbreviations GA gibberellic acid - 2iPA 2-isopentenyladenosine - Stage A heart-shaped embryo - siage B middle cotyledonary embryo - Z zeatin - Z₉G zeatin glucoside - Zr Zeatin riboside     

Biological control of arrowhead scale,Unaspis yanonensis,by parasitic wasps introduced from the People's Republic of China

In 1980, the authors went to the People's Republic of China in search of the natural enemies of the arrowhead scale,Unaspis yanonensis Kuwana, and succeeded in collecting 2 species of parasitic wasps of this pest namelyPhyscus fulvus Compere & Annecke andAphytis yanonensis DeBach & Rosen. The development, reproduction and longevity ofAphytis yanonensis were studied under constant temperature conditions. The number of generations a year was estimated to be from 10 to 12. Reproduction ofPhyscus fulvus is arrhenotokous, and development from egg to adult required 25 days at 25°C and 33 days at 20°C. The number of generations was estimated at 4 to 5 a year. Both parasites were released in several citrus groves in the Shizuoka Prefecture, of central Japan during June and July, 1981. They successfuly hibernated and each stage of development was found from March to December, 1982. They are considered to be potentially highly effective against the arrowhead scale. We have succeeded in establishing bothAphytis yanonensis andPhyscus fulvus in several districts in the Shizuoka Prefecture.

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Résumé Les auteurs se rendirent en 1980 en République Populaire de Chine pour entreprendre une recherche sur les ennemis naturels deUnaspis yanonensis Kuwana et découvrirent 2 espèces d'hyménoptères parasites de la cochenille. Les ennemis naturels introduits furentPhyscus fulvus Compere & Annecke etAphytis yanonensis DeBach & Rosen. La croissance, la reproduction et la longévité deA. yanonensis furent étudiées à température constante. Le nombre de générations annuelles a été estimé entre 10 et 12. La reproduction deP. fulvus est de type arrhénotoque et la durée du développement de l'œuf à l'adulte, nécessite 25 jours à 25°C et 33 jours à 20°C. Le nombre de générations annuelles a été estimé à 4 ou 5. Les 2 parasites ont été libérés dans plusieurs plantations d'agrumes de la préfecture de Shizuoka, région centrale du Japon pendant les 2 mois de juin et juillet 1981 et ont hiverné avec succès. Chaque stade de développement a été observé de mars à décembre 1982. Ces parasites sont considérés comme des ennemis naturels hautement efficaces de la cochenille. Nous avons introduit avec succèsAphytis yanonensis etPhyscus fulvus en provenance de Chine dans diverses régions de la préfecture de Shizuoka, partie centrale du Japon.

     

Contest and scramble competitions in two bruchid species,Callosobruchus analis andC. phaseoli (Coleoptera: Bruchidae)

Larval competition between contest and scramble strategists was investigated using the two bruchid species, C. analis (contest species) and C. phaseoli (scramble species) with two different sized mung beans (large and small beans). In both sized beans, the adult emergences of each species dependen on total density of the initial larval densities of the two species and the ratio of the two densities. The emergence of one species was suppressed by the existence of the other species when the initial larval density per bean of the former species was less than that of the latter one. There were many cases in which both C. analis and C. phaseoli emerged from one bean in large beans, but such cases were quite rare in small beans. C. analis performed interference behavior only at late larval stages, whereas C. phaseoli was superior in exploitative competition all through their larval stages. These, combined with the niche segregation inside a bean, are throught to be the major factors of observed density- and frequency-dependent competition results. Based on the above experimental results, long-term competition results between the contest and scramble species were predicted.     

Immunocytochemical localization of cathepsins B,H, and L in the rat gastro-duodenal mucosa

Summary Cathepsins B, H, and L are representative cysteine proteinases in lysosomes of a large variety of cells. Previous immunochemical studies indicated the presence of these enzymes also in the gastrointestinal wall. Using specific antisera, the cellular and subcellular distribution of cathepsins B, H, and L in rat gastric (oxyntic and pyloric part) and duodenal mucosa was investigated by light and electron microscopical immunocytochemistry. The subtypes of cathepsins were distributed differently in the cellular constituents of the epithelia: Cathepsin B was localized to lysosomes of all cells except goblet cells. Cathepsin H was found predominantly in gastric parietal cells (lysosomes) and in secretion granules of pyloric gastrin and duodenal cholecystokinin cells. Cathepsin L immunoreactivities were weak and restricted to a minority of cells (gastric mucous cells, enterocytes). Interstitial cells of the lamina propria immunoreactive for cathepsins H and L were identified as macrophages. The present findings suggest a dual function of cathepsins in the gastro-duodenal mucosa. They (1) cleave enzymatically proteins and peptides ingested in lysosomes, and (2) they may be involved in the processing of biologically active peptides (enteric hormones) from their precursor proteins.     

Application of immobilized lipase to regio-specific interesterification of triglyceride in organic solvent

Summary Lipase from Rhizopus delemar was immobilized by entrapment with photo-crosslinkable resin prepolymers or urethane prepolymers or by binding to various types of porous silica beads. The immobilized lipase preparations thus obtained were examined for their activity in converting olive oil to an interesterified fat (cacao butter-like fat), whose oleic acid moieties at 1- and 3-positions were replaced with stearic acid moieties, in the reaction solvent n-hexane. Although all of the immobilized preparations exhibited some activity, lipase adsorbed on Celite and then entrapped with a hydrophobic photo-crosslinkable resin prepolymer showed the highest activity, about 75% of that of lipase simply adsorbed onto Celite. Entrapment markedly enhanced the operational stability of lipase.Dedicated to Professor H. Holzer, Freiburg University, on his 60th birthday (June 13, 1981)     

Duct,exocrine, and endocrine components of cultured fetal mouse pancreas

Summary Twenty to twenty-two days postcoitum mouse fetal pancreas organ bits were cultured on the dermal surface of irradiated pigskin as a substrate. The medium used for long term culture consisted of Eagle’s Minimum Essential Medium with the addition of 10% bovine serum, 0.02 U/ml insulin, 0.025 μg/ml glucagon, 3.63 μg/ml hydrocortisone, 100 μg/ml soybean trypsin inhibitor or 10⁻⁸ M atropine. When the medium lacked trypsin inhibitor or atropine but contained the three hormones, the pigskin support began to be destroyed after 2 to 4 wk in culture. Thereafter, the cultured cells could not grow and survive on the digested pigskin. When 10⁻⁶ M atropine was added to the medium, amylase secretion from cultured cells and destruction of pigskin were inhibited completely but pancreas cells could not grow or survive. In contrast, 100 μg/ml soybean trypsin inhibitor or 10⁻⁸ M atropine permitted cell growth, permitted amylase secretion from the cultured acinar cells, and prevented the destruction of pigskin. Under these conditions pancreas cells migrated or grew or both from the organ bits onto the surface of the pigskin dermis and organoid aggregations formed. Hydrocortisone was needed to permit growth for more than 2 wk. Glucagon and insulin had additive effects. Light and electron microscopic observations indicated the culture of at least five kinds of cells, i.e., duct, acinar, centroacinar, endocrine, and mesenchymal. The majority of cultured cells were duct cells and acinar cells. There were few mesenchymal cells. Mouse pancreas cells were cultured for at least 12 wk by this method. This investigation was supported by PHS Grant CA 30220 awarded by the National Cancer Institute, DHHS, Grant 1203M awarded by the Council for Tobacco Research, Inc., and Grant RD-65 (for equipment) awarded by the American Cancer Society. Nude mice were provided by Dr. Wendall M. Farrow of Life Sciences, Inc., Resource Laboratory N01, CP6-1005 of the National Cancer Institute.     

Chirality of the acyl group of β-hydroxy-β-methylglutarylhydroxyabscisic acid

The β-carbon of the acyl group of β-hydroxy-β-methylglutarylhydroxyabscisic acid was shown to possess R-configuration by HPLC analysis of the reduced product.