Metabolic activation and hepatotoxicity. Toxicity of bromobenzene in hepatocytes isolated from phenobarbital-and diethylmaleate-treated rats.

Hepatocytes freshly isolated from diethylmaleate-treated rats exhibited a markedly decreased concentration of reduced glutathione (GSH) which increased to the level present in hepatocytes from nontreated rats upon incubation in a complete medium. When bromobenzene was present in the medium, however, this increase in GSH concentration upon incubation was reversed and a further decrease occurred that resulted in GSH depletion and cell death. This was prevented by metyrapone, an inhibitor of the cytochrome P-450-linked metabolism of bromobenzene. Bromobenzene metabolism in hepatocytes was accompanied by a fraction of metabolites covalently binding to cellular proteins. The size of this fraction, relative to the amount of total metabolites, was increased in hepatocytes isolated from diethylmaleate-treated rats and in hepatocytes from phenobarbital-treated rats incubated with bromobenzene in the presence of 1,2-epoxy-3,3,3-trichloropropane, an inhibitor of microsomal epoxide hydrase which, however, also acted as a GSH-depleting agent. In addition, the metabolism of bromobenzene by hepatocytes was associated with a marked decrease in various coenzyme levels, including coenzyme A, NAD(H), and NADP(H). Cysteine and cysteamine inhibited the formation of protein-bound metabolites of bromobenzene in microsomes, but did not prevent bromobenzene toxicity in hepatocytes when added at higher concentrations to the incubation medium (containing 0.4 mm cysteine). Methionine, on the other hand, did not cause a significant effect on bromobenzene metabolism in microsomes and prevented toxicity in hepatocytes, presumably by stimulating GSH synthesis and thereby decreasing the amount of reactive metabolites available for interaction with other cellular nucleophiles. It is concluded that, in contrast to hepatocytes with normal levels of GSH, hepatocytes from diethylmaleate-treated rats were sensitive to bromobenzene toxicity under our incubation conditions. In this system, bromobenzene metabolism led to GSH depletion and was associated with a progressive decrease in coenzyme A and nicotinamide nucleotide levels and a moderate increase in the formation of metabolites covalently bound to protein. Methionine was a potent protective agent which probably acted by enhanced GSH synthesis via the formation of cystathionine.     

The audibility of frog choruses to migrating birds

Breeding choruses of Hyla crucifer and H. versicolor are loud enough to be audible to migrating birds up to at least 1 km from their source, both vertically and horizontally, provided that no large obstacles intervene. During May in south-eastern New York State sound pressure levels (A weighting) at altitudes of 200 to 965 m and slant ranges from the frogs of 225 to 1020 m varied from 28 to 52 dB SPL.     

Tension headaches

In order to assess the effectiveness of verbal relaxation instructions and EMG relaxation training on the relief of tension headaches, 18 medically documented tension-headache sufferers were randomly assigned to one of three conditions:(1) Jacobson-Wolpe autogenic-relaxation training,(2) EMG relaxation training, or(3) EMG relaxation training combined with Jacobson-Wolpe autogenic instructions. The two EMG-assisted groups showed significantly better results, compared to the verbal group, in terms of reduction of headache activity and the rate at which reduction took place. Thus, EMG-assisted relaxation procedures seem the treatment method of choice for relief of tension headaches.     

Chromosome numbers in some North American species of the genus Cirsium. III. Western United States,Mexico, and Guatemala

New data are presented on chromosome numbers for 36 species, two varieties, and two hybrids ofCirsium (Compositae). These include first reports forC. rhothophilum (2n = 34),C. andrewsii (2n = 32),C. crassicaule (2n = 32),C. quercetorum (2n= 32, 112),C. pascuarense (2n= 32),C. douglasii var.canescens (2n = 30, 34),C. hydrophilum (2n = 32),C. neomexicanum (2n = 30),C. cymosum (2n = 30, 34),C. acantholepis (2n= 34),C. radians (2n = 34), C.grahami (2n = 32),C. nigriceps (2n = 36),C. andersonii (2n= 32, 64),C. anartiolepis (2n = 34), andC. subcoriaceum (2n= 34). The published data on chromosome numbers of Eurasian and AmericanCirsium are summarized. In Eurasia, speciation has taken place primarily at the diploid level but is occasionally reinforced by polyploidy. The ancestral base number of 17 has been preserved in almost all species, and there is little evidence that reduction in chromosome number has played a significant role in speciation. In America speciation has proceeded exclusively at the diploid level, but the ancestral genome of 17 chromosomes has been retained in only about half of the species examined. In the remaining species, restructuring of the genome has occurred resulting in a reduction in number from 17 to 9 in extreme cases. Polyploidy, when seen, is of no significance. It is suggested that all species with greatly reduced numbers may represent products of a single reduction series.     

A Suppressor Gene Involved in Chemical Induction of Conjugation in PARAMECIUM AURELIA

A dominant gene Su(kau-2) partially suppresses the effect of an apparently unlinked recessive kau-2. Gene kau-2 blocks chemical induction of conjugation in Paramecium aurelia syngen 8 when solutions of acriflavine + either KCl or MgCl-2 are used. Wild-type cells are induced to conjugate in either solution. When cells homozygous for kau-2 also have gene Su(kau-2), they are still uninducible in the solution containing KCl, but become inducible in the MgCl-2 solution. Analysis of the concentrations of solutions which are effective in induction of conjugation of various genotypes shows that the action of Su(kau-2) is not a simple restoration of wild-type phenotype since certain novel features of the suppressor can be seen. Analysis of the duration of ciliary reversal of various genotypes suggests that one necessary step in chemical induction of conjugation is a certain magnitude of depolarization of the surface membrane of the cell.     

Induction of base-pair substitution and prameshift mutations in wild-type and repair-deficient strains of Salmonella typhimurium by the photodynamic action of methylene blue

Induction of back mutations to prototrophy by methylene blue (MB)-sensitized photodynamic (PD) treatment has been studied in wild-type and repair-deficient strains of Salmonella typhimurium carrying either the base-pair substitution mutation hisG46 or the frameshift mutation hisD3052. We found that reversion of the hisG46 mutation was increased in a strain carrying a uvrB deletion and decreased in a strain carrying a recA-type mutation. Reversion of the hisD3052 (frameshift) mutation, on the other hand, was decreased in both uvrB deletion and recA-type strains. The former results are consistent with the hypothesis that the majority of MB-sensitized PD-induced base-pair substitution mutations arise by a mechanism similar to that currently believed to be involved in UV mutagenesis. The latter results suggest that PD-induced frameshift mutations may arise in some other way, and two possible mechanisms involving sequential action of the excision repair and recombinational repair pathways are considered.     

Bacteriological activity in unfiltered calf sera collected for tissue culture use

Summary Bacteriological tests were made on 24 lots of unfiltered calf serum collected for subsequent use as a component of tissue culture media. The examination included the isolation and identification of bacteria, assay of phages, and demonstration of endotoxin material. Only Gram-positive bacteria were isolated and 96% of the sera were contaminated with bacteria. The prevalent strains of bacteria found wereBacillus species and streptococci and 63% of the sera coagulatedLimulus amebocyte lysate. More than 90% of the lots contained phages demonstrable with the C-3000 strain ofEscherichia coli. Only one lot of the serum was found to be free from bacteria, phages, and endotoxin by the tests used.