Rhizobium japonicum mutants that are hypersensitive to repression of H2 uptake by oxygen.

The synthesis of an H2 oxidation system in free-living Rhizobium japonicum wild-type strain SR is repressed by oxygen. Maximal H2 uptake rates were obtained in strain SR after derepression in 11 microM or less dissolved oxygen. Oxygen levels above 45 microM completely repressed H2 uptake in strain SR. Five R. japonicum mutant strains that are hypersensitive to repression or H2 oxidation by oxygen were derived from strain SR. The mutants were obtained by screening H2 uptake-negative mutants that retained the ability to oxidize H2 as bacteroids from soybean nodules. As bacteroids, the five mutant strains were capable of H2 oxidation rates comparable to that of the wild type. The mutants did not take up H2 when derepressed in 22 microM dissolved oxygen, whereas strain SR had substantial activity at this oxygen concentration. The O2 repression of H2 uptake in both the wild-type and two mutant strains, SR174 and SR200, was rapid and was similar to the effect of inhibiting synthesis of H2 uptake system components with rifampin. None of the mutant strains was able to oxidize H2 when the artificial electron acceptors methylene blue or phenazine methosulfate were provided. The mutant strains were not sensitive to killing by oxygen, they took up O2 at rates similar to strain SR, and they did not produce an H2 uptake system that was oxygen labile. Cyclic AMP levels were comparable in strain SR and the five mutant strains after subjection of the cultures to the derepression conditions.     

A spin labeling study of the effects of inorganic ions and pH on the conformation of spectrin

The structure of spectrin from human erythrocytes has been investigated by the EPR technique measuring the mobility of the protein spin label, 4-maleimido-2,2,6,6-tetramethylpiperidinooxyl. Conformational changes in the protein induced by variation of the concentrations of NaCl, Na2SO4, KCl, CaCl2 and MgCl2 and of pH have been studied. It could be demonstrated that both Ca2+ and Mg2+ give rise to structural changes by binding to specific sites, whereas the monovalent cations (K+, Na+) seem to act via ionic strength. A model is used to correlate the spin label mobility with the radius of the protein. In the Ca2+- and Mg2+-binding experiments, the decrease in the spin label mobility has been interpreted on the basis of the theory of multiple chemical equilibria. These experiments have been compared with EPR spectra measured at different pH values. The results support the model in that binding of H+, Ca2+ or Mg2+ reduces the charges located on the protein surface: the 'discharging' reduces the repulsive forces on the surface of the molecule and consequently, the protein contracts in discrete steps.     

Influence of subcellular fractions of mammalian testes on the mutagenic activity of nitrofurans toward Escherichia coli; presence of a co-mutagen-like factor.

The activation of nitrofurans to mutagenic intermediates by testicular tissue was investigated. AF-2 and nitrofurazone were tested in a microsomal suspension assay with strain E. coli K-12 343/113 as indicator and subcellular fractions from rabbit testes. Different mutation patterns were observed in the presence or absence of testicular homogenate, indicating the presence of different mutagenic intermediates. The frequency of arg+ reversion increased proportionally to the homogenate concentration suggesting that the nitrofurans were activated by testicular components to intermediates that induced base-pair substitutions. Other experiments showed that a component of low molecular weight, present in the soluble fraction of homogenates of testes from rabbits, rats and monkeys, was responsible for the increased mutation frequency. It is concluded that this "co-mutagen-like" factor either alters the metabolism of nitrofurans in E. coli and/or promotes the formation of base-pair substitution-type mutations. This direct interaction between a nonenzymic component of mammalian testes and the mutation induction/expression process in E. coli suggests the role of co-mutagen-like factors in the sensitivity of testes to nitrofurans.     

Effect of canine surfactant protein (SP-A) on the respiratory burst of phagocytic cells

Cells obtained from bronchoalveolar lavage, or neutrophils of peripheral blood of dog, were incubated with the canine surfactant-associated protein A (SP-A). A significant decrease of the production of Superoxide anion was observed after subsequent stimulation with phorbol-12-myristate-13-acetate (PMA) as measured by the lucigenin-dependent chemiluminesence (CL). Several other proteins used for control experiments did not decrease lucigenin-dependent CL, indicating a specific effect of SP-A on phagocytes. Treatment of SP-A with collagenase prior to incubation with neutrophils destroyed the depleting effect on oxygen radical production of PMA-stimulated cells. We propose that SP-A acts as a regulatory factor of the respitatory burst of alveolar macrophages and neutrophils in the lungs. The inhibitory effect of SP-A is down-regulated by collagenase released from stimulated alveolar macrophages.     

Spatial distribution of resting stages,rate of emergence from diapause and times to adulthood and to the appearance of the first clutch in 3 species of cyclopoid copepods

The spatial distribution of dormant copepodids of 3 species of cyclopoid copepods — Cyclops vicinus, Mesocyclops leuckarti and Thermocyclops crassus — was studied in 4 small lakes in South Germany. The rate of emergence from diapause and times from the resting stage to adulthood and from adulthood to the appearance of the first clutch was studied at 4 constant temperatures (5, 10, 15, 20 °C) in the laboratory. Resting stages of C. vicinus were always concentrated in the deepest parts of the lakes and were found relatively deep in the mud. M. leuckarti- and T. crassus-copepodids preferred shallow areas in deep lakes but were concentrated in the deep areas in shallow lakes. Copepodids of both species were always concentrated in the uppermost mud layers.Rate of emergence from diapause was strongly temperature-dependent. At high temperatures (20 °C) copepodids of all species under study emerged within 2 weeks. At lower temperatures C. vicinus copepodids showed the highest rate of emergence. At 5 to 10 °C only few M. leuckarti- and T. crassus-copepodids had emerged after the investigation period (7 weeks). Both C. vicinus and T. crassus showed the highest rate of emergence at the natural end of diapause but even at that time only few T. crassus-copepodids emerged at 5 °C. Times to adulthood at 5 °C were shortest in C. vicinus. At higher temperatures this species was passed by M. leuckarti. Times from adulthood to the appearance of the first clutch at 5–15 °C were shortest in C. vicinus. T. crassus produced no clutch at 5 and 10 °C.     

The effect of temperature on the development,reproduction, and longevity of two common cyclopoid copepods — Eucyclops serrulatus (Fischer) and Cyclops strenuus (Fischer)

The embryonic and postembryonic development times, time interval between clutches, and longevity of two common cyclopoid copepods — Eucyclops serrulatus and Cyclops strenuus — were studied at constant temperatures (5°, 10°, 15°, 20° and 25 °C), using algae and protozoans as food. Both, E. serrulatus and C. strenuus showed a considerable temperature tolerance and adaptability. Embryonic development times were similar in both species but postembryonic development times were shorter in E. serrulatus. Time intervals between successive clutches were relatively variable at all temperatures in both species. Longevity was shorter in E. serrulatus than in C. strenuus. The embryonic and postembryonic development of E. serrulatus was short relative to that of littoral copepods; the development of C. strenuus was similar or shorter than that of other species of the genus Cyclops.     

Nickel accumulation and storage in Bradyrhizobium japonicum.

Hydrogenase-derepressed (chemolithotrophic growth conditions) and heterotrophically grown cultures of Bradyrhizobium japonicum accumulated nickel about equally over a 3-h period. Both types of cultures accumulated nickel primarily in a form that was not exchangeable with NiCl2, and they accumulated much more Ni than would be needed for the Ni-containing hydrogenase. The nickel accumulated by heterotrophically incubated cultures could later be mobilized to allow active hydrogenase synthesis during derepression in the absence of nickel, while cells both grown and derepressed without nickel had low hydrogenase activities. The level of activity in cells grown with Ni and then derepressed without nickel was about the same as that in cultures derepressed in the presence of nickel. The Ni accumulated by heterotrophically grown cultures was associated principally with soluble proteins rather than particulate material, and this Ni was not lost upon dialyzing an extract containing the soluble proteins against either Ni-containing or EDTA-containing buffer. However, this Ni was lost upon pronase or low pH treatments. The soluble Ni-binding proteins were partially purified by gel filtration and DEAE chromatography. They were not antigenically related to hydrogenase peptides. Much of the 63Ni eluted as a single peak of 48 kilodaltons. Experiments involving immunoprecipitation of 63Ni-containing hydrogenase suggested that the stored source of Ni in heterotrophic cultures that could later be mobilized into hydrogenase resided in the nonexchangeable Ni-containing fraction rather than in loosely bound or ionic forms.     

Analysis of cellular interactions in density-dependent inhibition of 3T3 Cell proliferation

Subpopulations of different proliferative status are determined during cell-density dependent proliferation of 3T3 cells. From these data the probability of conversion of proliferative to quiescent cells is derived and found to correlate well with published data on binding of growth-inhibiting factors secreted from growth-inhibited cells.Based on material presented at the Symposium Intercellular Communication Stuttgart, September 16–17, 1982     

Anti-tumor effects of an antiserum raised in syngeneic mice to a tumor-specific T suppressor factor

Summary DBA/2 mice inoculated with either cells from the syngeneic P815 tumor or tumor cell membrane extracts develop T suppressor cells which suppress the in vitro generation of cytotoxic T lymphocytes with specificity for the tumor. A soluble suppressor factor with similar properties can be isolated from suppressor cell-enriched populations. It can be highly purified by appropriate immunoadsorption. Antisera to this suppressor factor raised in either DBA/2 or C57BL/6 mice can specifically absorb out suppressor factor and eliminate suppressor cells in the presence of complement. The in vivo effects of these antisera were tested for their ability to modulate the growth of P815 tumors in DBA/2 mice. It was found that the antiserum raised in syngeneic (DBA/2) but not allogeneic (C57BL/6) mice was able to significantly slow the rate of tumor growth and to prolong survival in treated mice. The antiserum was effective in this way only if it was administered early in the course of tumor growth. It was shown that this effect was not attributable to the presence in the serum of antibodies directed to antigens present on P815 cells, and it therefore appears to be due to interference with the function of T suppressor cells arising early in the immune response to the tumor cells.