Isolation and Characterization of a Novel Thermophilic, Freshwater Methanogen

A novel thermophilic, coccoid methanogen isolated from nonthermal freshwater sediments is described. Hydrogen plus carbon dioxide and formate were substrates for methanogenesis, and methane production was stimulated by yeast extract, Casamino Acids, and tryptose. Growth also occurred autotrophically. Elevated levels of sodium chloride were not required for maximum growth and were inhibitory above 2%. The minimum doubling time occurred at 57°C, and the upper and lower limits for methane production were 62 and 26°C, respectively. The optimum pH for growth was between 7.0 and 7.5. Inhibitory antibiotics included metronidazole, anisomycin, chloramphenicol, and lasalocid. Colonies were circular, dark yellow, shiny, and convex with entire edges. Cells were 1 to 2.5 μm in diameter, nonmotile, occurring singly or in pairs, and fimbriated. Cells were lysed by pronase or trypsin digestion, glass-distilled water, and 1% sodium dodecyl sulfate. Electron micrographs of thin sections showed a monolayered cell wall ca. 20 nm thick. The DNA base ratio was 49.2 mol% guanine plus cytosine. The whole cell protein pattern differed from that of other named coccoid methanogens.     

High phosphate requirement for oxidative phosphorylation and low affinity for phosphate transport in newborn rat liver mitochondria

Rat liver mitochondria are not fully functional at birth. The relationship between this deficiency and the affinity for phosphate, in oxidative phosphorylation or in phosphate transport, have been studied.The phosphate concentration necessary to observe maximal rate of succinate oxidation in the presence of ADP was higher for newborn than for adult rat liver mitochondria. After preincubation of newborn rat liver mitochondria with ATP, the rate of succinate oxidation in the presence of ADP increased with phosphate concentration similarly for newborn and adult rat liver mitochondria. The maximal rate of phosphate-acetate exchange, which is an indirect measure of the rate of phosphate transport across the mitochondrial membrane, was not significantly different for adult and newborn rat liver mitochondria. On the contrary the apparent affinity for phosphate was about ten-fold lower for newborn than for adult mitochondria.     

Chloroherpeton thalassium gen. nov. et spec. nov., a non-filamentous,flexing and gliding green sulfur bacterium

A flexing and gliding green sulfur bacterium has been isolated from marine sources off the North East coast of the USA. Chloroherpeton thalassium is an obligate phototroph, and requires CO₂ and S^2- for growth; some organic acids can contribute to cell carbon, and N₂ may be fixed. The cells contain typical chlorosomes, and gas vesicles may be present. Bacteriochlorophyll c is the main light harvesting pigment, and a small quantity of bacteriochlorophyll a is also present. Over 80% of the carotenoid is -carotene. DNA base composition of the isolates ranges from 45.0–48.2 mol% G+C.In memory of R. Y. Stanier     

Phytohormones,Rhizobium mutants, and nodulation in legumes. VI. Metabolism of zeatin riboside applied via the tips of nodulated pea roots

[³H]zeatin riboside was supplied in physiological quantities to pea (Pisum sativum L. cv Greenfeast) plants by replacing the root tip with a small vial containing [³H]zeatin riboside, to simulate the normal supply of cytokinin. Radioactivity was transported to the root nodules. Analysis by two-dimensional thin layer chromatography revealed that little³H remained as zeatin riboside in root or nodule tissue at the end of the labeling period (2, 5, or 8 d) and suggested that the following compounds were metabolites of [³H]zeatin riboside: zeatin, adenosine, adenine, the O-glucosides of zeatin and zeatin riboside, nucleotides of adenine and zeatin, and the dihydro-derivatives of many of these compounds.The O-glucosides (and in particular, O--D-glucopyranosyl-9--D-ribofuranosylzeatin) appeared to be more prominent metabolites in the effective nodules formed by strain ANU897 than in the ineffective nodules produced by strain ANU203. However, no other appreciable differences were detected between effective and ineffective nodules in their metabolism of zeatin riboside. There were few marked differences between root and nodule tissue; however, in some experiments, the nodules contained a higher proportion of O-glucoside metabolites, and generally root tissue contained a greater proportion of zeatin and/or dihydro-zeatin, zeatin riboside and/or dihydrozeatin riboside, adenine and the nucleotides of zeatin and adenine, as metabolites.     

The milk of the fur-seal, Arctocephalus tropicatis gazella; in particular the composition of the aqueous phase

Milk and plasma samples from the Southern fur-seal (Arctocephalus tropicalis gazella) , collected from animals shot between 0 and 26 days post-partum , have been analysed. The aqueous phase of milk (ultracentrifugate) was hyperosmotic to plasma by about 35 m-osmol/kg water but contained no lactose. The osmolality of milk could be accounted for by the ionic content, of which sodium and potassium (in approximately equal amounts) were the main cations. The fat content of the milk was very high (approximately 40%), except in very early lactation (0 and 1 day post-partum). The concentrations of chloride, calcium, magnesium, phosphate and citrate in the aqueous phase are given. Major differences between the composition of fur-seal milk and that of other mammals are pointed out, and discussed in relation to water conservation, energy and fat supply, and prolonged milk storage in this snecies.     

Production of erythropoietin in vitro: A review

Summary The in vitro production of the important regulatory of erythropoiesis, erythropoietin (Epo), is reviewed. It is concluded that it is possible to produce almost routinely small quantities of Epo in tissue culture. Although such procedures offer the potential to provide large quantities of the hormone for clinical use, the optimum culture conditions and mechanisms for triggering Epo production have yet to be resolved. This work was supported by Grants No. 74444 from The John A. Hartford Foundation, and HL 10567 from the National Institutes of Health.     

Formation of junctions and cell sorting in aggregates of chick and mouse cells

The frequency of desmosome formation was examined in aggregates of old cells, which form many junctions, combined with young cells, which form few. Cells of chick corneal epithelium and mouse epidermis, which can be distinguished morphologically, were combined. Desmosomes between these cell types are stable. Further, young cells make more desmosomes than they otherwise would on those surfaces adjoining old cells. Desmosomes increase in number in aggregates while cell sorting is occurring. Cells consistently sort, with those which form most desmosomes lying internally. Gap junctions and intermediate junctions are also present, but are uncommon. A carbohydrate cell-surface coat has regenerated by the time desmosome formation starts. The possible relation of desmosome formation to cell sorting is discussed.     

Biosynthesis of cat hemoglobins A and B: Amino terminal acetylation of the β chain of HbB

Acetylation of the amino terminal serine of the β chains of cat HbB occurs during synthesis of hemoglobin in a mRNA dependent rabbit reticulocyte lysate protein synthesizing system in the presence of acetyl-CoA and cat reticulocyte mRNA. Both of the major cat hemoglobins, the nonacetylated HbA and acetylated HbB, are synthesized efficiently in the rabbit lysate system. The acetylation of HbB-β chains occurs during the biosynthesis of these proteins. Radioautography of tryptic peptide maps reveals that acetylation occurs specifically at the amino terminal serine of HbB-β globin, and not on HbA-β globin or on the α chain common to both hemoglobins A and B. Because of the similarity of the structures of HbA-β and HbB-β globin, it is suggested that the amino terminal residue determines whether the peptide chain is recognized for acetylation by a ribosomal acetyltransferase.     

Nutritional and karyotypic characterization of a haploid cell culture ofDaucus carota

Summary The purpose of this study was to optimize growth conditions for a strain of haploid carrot callus and to follow its karyotypic changes in a long span of time. The strain has been maintained in liquid suspension since September 1977. It has remained predominantly haploid in its karyotype since that time. The original explant was initiated and subsequently subcultured in Gamborg's B5 medium. The components of the B5 medium were omitted one at a time and sequentially added back to determine their minimum, optimum, and maximum nontoxic concentrations. These changes were made in the original formula: the addition of an organic buffering agent and an increase in the iron and other micronutrient concentrations. Using this slightly modified B5 medium, we assessed the effect on growth by single additions of amino acids, different carbon sources, growth regulators, and vitamins. No improvement in plating efficiency resulted from addition of any of these compounds. We conclude that there are factors limiting the plating efficiency of the haploid cells other than these tested, or that single additions will not make a discernible difference, or that growth promoting factors cannot be exogenously supplemented to cultured cells.