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.     

Pollen frequencies reflect vegetation patterns in a great basin (U.S.A.) mountain range

Five indices are used to quantify the relationship between vegetation and pollen in a mountain range in the arid Great Basin. Computations are based on vegetation coverage and pollen percentages from 63 stands. Association is a measure of whether the presence of the pollen type in a surface sample is an indication of the presence of the parent plant in the local vegetation. Over-representation and under-representation measure tendencies for pollen to occur where the parent plants are absent and vice versa. The correlation coefficient measures the relationship between plant and pollen in stands where both are present. Twenty-nine trees, shrubs, and herbs accurately reflect local vegetation conditions. A percentage diagram shows elevational trends in abundant pollen types. Regional pollen types are used to compute the accumulation rate of pollen in the surface samples. A diagram of pollen accumulation rates shows trends similar to those shown in the percentage diagram. The moss polsters used in this study may collect pollen over a fifteen-year interval.     

Monoamines and lordosis in the female guinea pig.

Serotonin and dopamine antagonists have been reported to induce lordosis in estrogen-primed spayed rats in the absence of progesterone. In the present study with guinea pigs, however, no such effect was found with any of the drugs tested, which include methysergide, parachlorophenylalanine (PCPA), haloperidol, and amphetamine. When given with progesterone, methysergide slightly decreased latency to heat, whereas PCPA substantially reduced lordotic response. Methysergide did not alter the inhibitory phase of progesterone action.     

High performance liquid chromatography of pharmacologically active amines and peptides in biological materials

Precise and quantitative reversed-phase high performance liquid chromatographic (HPLC) procedures are described which can be used in biogenic amine and neuropeptide research. The amine procedure was applied to various pharmacological matrices including plasma, heart tissue and brain. The use of peptide HPLC as an analytical tool for various neuropeptides is illustrated by studies on des-tyrosine-gamma-endorphin (DT gamma E) metabolism in the brain and the stability of an ACTH (ORG-2766) analogue during a chronic infusion in rats. The power of HPLC as a research tool in peptide pharmacology is described, discussed and demonstrated as an aid in the understanding of the pharmacological effects of exogenous peptides and the function of the brain.     

N-demethylation of lergotrile by Streptomyces platensis.

Thirty-eight microorganisms were screened for their ability to produce metabolites of the semisynthetic alkaloid, lergotrile. A total of five microorganisms were found to biotransform lergotrile, and N-desmethyl lergotrile was detected as the principal metabolite with most organisms. Streptomyces platensis (NRRL 2364) appeared to form the metabolite in highest yield, and a preparative-scale conversion was accomplished with a recovered yield of 50%. Structure proof was accomplished with comparative thin-layer chromatography, mixed melting point, mass spectrometry, and remethylation to lergotrile.     

Calcium ion-stimulated phosphorylation of myelin proteins.

Myelin isolated from the central and peripheral nervous system contains a Mg2+-dependent protein kinase that catalyses phosphorylation of myelin-specific proteins. This phosphorylation is markedly stimulated by Ca2+ but not by cyclic AMP. Evidence was obtained that suggested an involvement of calmodulin-like protein in the stimulatory effects of Ca2+ on myelin phosphorylation.     

Functional group contributions to the partitioning of phenols between liposomes and water

The temperature dependency of the partitioning of p-alkylphenols and p-halophenols has been determined between dimyristoyl phosphatidylcholine liposomes and 0.15 M NaCl. Partition coefficients increased as a function of temperature below the endothermic phase transition temperature (T_c) of the phospholipid but decreased above this temperature. The transfer process was found to be entropy-dominated below and enthalpy-dominated above the T_c, although large negative entropy changes were observed. Regular changes in the thermodynamic functions, partition coefficients and functional group free energies occurred as a function of the alkyl chain length or size of the halogen substituent below but not above the T_c. This has tentatively been attributed to increased phenol-phospholipid interaction at the higher temperatures. The partitioning of p-fluorophenol behaved in a manner expected of fluorinated compounds, yielding relatively low partition coefficients, but it produced an additional effect of markedly lowering the T_c of dimyristoyl phosphatidylcholine. Good correlations of the partition coefficients in liposomes with those in bulk organic solvents and with molecular size of the solute have been obtained.     

Membrane-bound protein kinase activity in acetylcholine receptor-enriched membranes

Membrane protein phosphorylation may be a general regulatory mechanism mediating the response of cells to exogenous metabolic and physical signals. We have determined that the membrane-bound acetylcholine receptor is the major substrate phosphorylated in situ by a nearby membrane protein kinase. Moreover, these same membranes also contain phosphoprotein phosphatase activity which dephosphorylates the membrane-bound receptor. These findings suggest that reversible phosphorylation of the actylcholine receptor may be critical for receptor function at the synapse. Therefore, it is necessary to define the properties of the enzymes which mediate this phosphorylation-dephosphorylation mechanism. In this report we describe the properties of the first component of this system, the membrane-bound protein kinase in receptor-enriched membranes from the electric organ of Torpedo californica. Only ATP is effective as a phosphate donor for this cyclic AMP-independent membrane kinase; GTP does not support phosphorylation of the receptor. Both casein and histone can also be phosphorylated by the membrane protein kinase, but casein is a better substrate. Although phosphorylation of the receptor appears to be regulated by cholinergic ligands and K+, casein phosphorylation is not specifically affected by these agents. Moreover, while phosphorylation of the acetylcholine receptor is maximal in receptor=enriched membranes, casein phosphorylation is similar in all membrane fractions prepared from the electric organ. Taken together, these findings suggest that the membrane protein kinase activity in receptor-enriched membranes is similar to most other membrane kinases. Therefore, the unique characteristics of membrane-bound acetylcholine receptor phosphorylation appear to be determined by the receptor and its availability as a substrate for the membrane kinase.