Growth of Escherichia coli A-l under aerobic conditions in an enriched medium with a total amount of 0.2 per cent glucose was biphasic and asparaginase II activity was detected after depletion of ammonia from the growth medium in the second phase of growth. Glucose was exhausted two hours before ammonia and three hours before asparaginase II activity was detected. The concentration of 3',5'-cyclic adenosine monophosphate was found to fluctuate when the dissolved oxygen in the medium reached a low level, when glucose and ammonia were exhausted, and when the cells entered the second stationary phase of growth. Culture tube studies of the growth of E_j_ coli A-l in three per cent nutrient broth with varied concentrations of ammonium chloride and potassium nitrate gave lower specific activity of asparaginase II when this was compared to that seen in three per cent nutrient broth alone. The addition of glucose to the same medium before asparaginase II activity was detected resulted in the production of acid by E. coli A-l with cessation of growth; however, addition after L-asparaginase synthesis had started did not affect the specific activity of the enzyme. The addition of ammonium chloride suppressed L-asparaginase synthesis, but addition after enzyme synthesis …

N,N'-methylenebisacrylamide (MBA) was studied because of its effectiveness in inducing heritable translocations in germ cells of male mice. The health impact of translocations was studied through anatomical analysis of the progeny of semisterile translocation carriers. As expected, the semisterility of translocation carriers resulted primarily from embryonic death during periimplantation stages due to unbalanced chromosome sperm segregants. Among conceptuses that survived to mid- and late-gestation stages, there was an increased incidence of developmental anomalies including fetal death and phenotypic defects. These abnormalities are associated with unbalanced chromosome complements that allow survival to the later stages of development.

A 17.1 Kb genetic element encoding for mercury resistance (OCT-Hg^r) was shown to translocate from its original location on the OCT plasmid to the resistance plasmid, RPl, in Pseudomonas putida. Analysis of RPl-Hg^r recombinant plasmids revealed that insertion of mercury resistance genes into RPl could occur at a variety of sites, with all recombinants having common EcoRI restriction fragments of 9.4, 3.8, 2.3, and 1.6 Kb, derived from the insertion. Hybridization analysis suggested the existence of extensive homology between this insertion and the prototypic mercury resistance transposon, Tn501, as well as the location of a similar merA sequence. Although the overall size was shown to be quite different from Tn501, striking physical similarities are shared between these two elements.

Chronic toxicity of zinc to Pimephales promelas was estimated by conducting replicate static and static-renewal short-term embryo-larval tests and static-renewal seven-day larval tests. The two test methods were highly reproducible. Daily renewal of test solutions had little effect on the toxicity of zinc, however, the stage of development at which exposure was initiated affected the sensitivity of the toxic endpoints measured. The most sensitive and reproducible endpoint in the embryo-larval tests was survival of viable (non-deformed) larvae and in the seven-day larval test was growth of the larvae, which was slightly more sensitive than the embryo-larval test endpoint. The estimated MATC of 0.18 and 0.15 mg/L mean total and mean soluble zinc, respectively, compared well with published results. Because of its advantages and similar sensitivity, the short-term embryo-larval test was recommended for estimating chronic toxicity.

The effects of external electric fields (EEF) on brain activity in anesthetized rats were studied. The field strengths used, 9 kV/m and 5 kV/m, both D.C. and A.C. (60Hz) were in the range of those measured beneath current overhead transmission lines. Brain activity was monitored from surface electrodes and from electrodes stereotaxically implanted in the posterior-lateral portion of the hypothalamus. It was found that 9 kV/m and 5 kV/m EEF's both D.C. and A.C. brought about statistically significant changes in hypothalamic activity, however, the effects were bi-directional, (i.e. increases and decreases). Only seven of the 60 animals exposed showed changes in the EEG recorded with surface electrodes. The data clearly indicate that (1) anesthetized animals do respond to a change in the external electric field around them, (2) the hypothalamus may contain special electro-receptors that, in turn, may alter various other physiological processes, and (3) the data indicates the need for further research to help government agencies to establish more adequate safety guidelines.

In a minimal medium supplemented with glucose and yeast extract, the optimum pH for the growth of C. poinsettias was found to be 7.5. The organism requires thiamine, biotin, and pantothenic acid for growth. No absolute requirement was found for any amino acid, purine or pyrimidine although an amino acid mixture was stimulatory. Casamino acids could be substituted for the synthetic amino acid mixture. Yeast extract provided an additional factor(s) necessary for maximal growth. The results suggest that the unknown factor found in yeast extract might be purified by a combination of solvent extraction, and adsorption and elution from charcoal.

National Water Quality Criteria intended to protect aquatic life and their uses from the adverse effects of pollutants may not be appropriate due to site-specific factors that alter chemical bioavailability. The Indicator Species Procedure may be used to derive site-specific criteria in order to account for differences in site-specific bioavailability. This procedure was implemented using zinc for three chemically different site (river) waters. The purpose of this study was to quantify the bioavailability of zinc in each site water and correlate results to water quality parameters and/or zinc speciation. Results demonstrated that national criteria for zinc accurately predicted the experimentally derived site-specific values within a factor of two when adjusted for water hardness. Particulate forms of zinc were shown to be biologically unavailable under conditions tested.

Neotoma micropus from Jack County, Texas, were studied over a 9-month period. Loci from blood and saliva were used to determine genetic variation within the population. Deviations from Hardy-Weinberg equilibrium were found at one locus. The average temporal F over all seven loci was 0.040. Genetic structuring was subtle, fluctuated on a seasonal basis, and was due to differential migration or predation on genotypes. Heterozygotes tended to move more than homozygotes, and a greater proportion of heterozygotes were lost from the population during each season. Genetic variation was maintained in the population by immigrant individuals. This differential in dispersal of genotypes fits current models of reorganization within the genome of populations.

The adenylate energy charge values of twenty bacteria isolated from soil and cultured in a medium consisting of soil and distilled water were determined by the luciferin-luciferase bioluminescense method. The purpose of this study was to examine the growth and energy charge values of these organisms in soil extract medium, and to determine what effect the addition of glucose has on their energy charge values. Three of the organisms employed in this study showed energy charge values similar to those reported for bacteria grown in enriched media. The remainder of the isolates demonstrated low energy charge values, and scant growth in the soil medium.

A new column, Dionex AS4A, (polystyrenedivinylbenzene matrix) used for the separation of ribonucleotides and deoxyribonucleotides for the first time, and previously used for ion analysis was found superior to conventional silica columns because it separates ribonucleotides and deoxyribonucleotides. Resolution of dGTP was not possible with the Dionex column and CTP and GDP often co-eluted. Using conventional silica columns, monophosphates separated from diphosphates and diphosphates from triphosphates. Using the new Dionex column resolves all three simultaneously. The Dionex column resolved nucleotides with sharper peaks than silica columns, and the longer its retention time the better was the resolution. This Dionex column is stable, with 80 runs possible without cleaning while resolving ribonucleotides and deoxyribonucleotides to the picomole level.

Mean salinity concentrations in the four reservoirs (Moss, Ray Hubbard, Texoma and Possum Kingdom) ranged from 0.2 ppt in Moss Lake to 2.01 ppt in Possum Kingdom Lake. Reservoir sodium and chloride concentrations were hypotonic to hypertonic to plasma levels in all species. Interspecific differences were seen in ionic concentrations within each reservoir. Total osmotic and sodium concentrations in carp, Cyprinus carpio, were correlated to their concentrations in the reservoirs. No such relationship was noted for chloride, potassium and calcium. A laboratory study indicated that fish collection by electroshock did not bias plasma ion concentrations. Exposures to wide variations in ionic concentrations did not appear to induce stress in the species studied.

The basis of this investigation is to characterize growth property differences in normal versus virally transformed cells. Using a temperature-sensitive mutant of Rous sarcoma virus, the cells' transformation state is regulated by the growth temperature; at 33°C the cells are transformed, while at 39°C the cells have normal characteristics. The morphology of NRK cells is elongated and fibroblastic; when transformed the cells are rounded. Normal cells grow to a monolayer and stop, while transformed cells grow to saturation densities greater than just a monolayer amount. Transformed cells can form foci when grown in mixture with normal cells. Normal cells must be in contact with the culture vessel in order to grow, but transformed cells lack anchorage dependence for growth.

Olfactory transduction mechanisms are best studied when specific blocking agents are available with which to modify cellular responses to odorant stimulation. This study is an electrophysiological investigation of functional group-specific olfactory acceptor mechanisms using ethylbromo acetate (EtBrAc), a World War I war gas which inhibits olfactory responses to odorant stimulation. The major findings of this investigation show that (1) vaporous EtBrAc is found to be a quick and effective inhibitor of electroolfactogram (EOG) responses to odorant stimulation, (2) isoamyl acetate is shown to protect EOG responses to a wide variety of odorants, inclusive of itself, from the inhibition effect of EtBrAc treatment, and (3) amine acceptor sites exist which are resistant to the inhibitory effects fo EtBrAc treatment.

A new procedure has been developed to concentrate the phosphofructokinase from muscle of Ascaris suum with minimum loss of activity. By utilizing this method, 50 ml fraction was concentrated to a final volume of 3 ml in about 1.5 h without loss in enzyme activity. The concentrated enzyme had a specific activity of 64 units per mg. Ascaris muscle-cuticle was incubated in 50 1M solutions of either acetylcholine, serotonin, y-aminobutyric acid, levamisole, or saline alone. Phosphate analysis of the isolated phosphofructokinase from each incubation revealed that the enzyme contained the following moles of phosphate per subunit: 2.9 (acetylcholine), 2.2 (serotonin), 2.0 (y-aminobutyric acid), 1.5 (levamisole), and 3.4 (salne alone). The present study did not establish a direct correlation between degree of phosphorylation and phosphofructokinase activity. Phosphofructokinase from muscle of Ascaris suum appears to contain several phosphorylation sites, and one of these sites is required to be phosphorylated in order for the enzyme to exhibit maximum activity under physiological conditions.

During four growing seasons, aquatic vascular plant production and distribution were studied in Pat Mayse Lake, Texas, a 2425 hectare oligo-mesotrophic reservoir. The dominant macrophyte population was Myriophyllum spicatum L. Growth rates and regrowth rates of mechanically harvested Myriophyllum beds were found to be dissimilar. Based on estimates of watermilfoil nutrient content, there were insufficient nutrients in the entire population to alter the trophic status of this reservoir should all of the nutrients be instantaneously released. Sediments were the primary nutrient (nitrogen and phosphorus) sink. Bank erosion and solids transport from the watershed appear to contribute most of the sediments and a lake-wide mean sedimentation rate of 2.5 cm/year was estimated from sediment trap and core sample data.

N^6,O^2' -Dibutyryl cyclic adenosine 3',5'-monophosphate (Bt_2cAMP) was investigated for its effects on various tissue culture cells infected with temperature-sensitive (ts) mutant, LA31 and Bratislava 77 (B77), a wild-type Rous sarcoma virus. Specifically, known parameters of transformation were investigated and a possible site of action has been tenably proposed. The drug Bt_2cAMP was found to have little effect on the transformation related properties of primary chick embryo fibroblasts (CEF) infected with either virus or normal rat kidney fibroblasts (NRK) infected with the wild-type B77-RSV. However, significant inhibition of the transforming properties in NRK infected with the ts mutant LA31 (LA31-NRK) were reported at the permissive temperature 33 degrees centigrade (33 C).

Carotenoid pigments were extracted from Corynebacteriuma poinsettiae (wild type) ATCC 9682, and from 108 mutants obtained by exposure of a streptomycin resistant strain of C. poinsettiae to ultra-violet light irradiation and N-methyl- N'-nitro-N-nitrosoguanidine. The pigments were characterized by their absorption maxima, Rf-values, and partition ratios in petroleum ether and methanol. Thin layer chromatography was used to compare pigments of the wild type with those of the mutants. Possible biosynthetic pathways in carotenoid synthesis of the wild type were postulated on the basis of the observed genetic blocks. Mutants were found which suggested the existence of a linear pathway in carotenoid synthesis from the aliphatic C4 0 molecule to the bi-cyclic C50-diol. Other mutants suggested possible alternative pathways in the biosynthesis of these pigments or the presence of intermediates not detectable by thin layer chromatography.

Responses of periphyton and phytoplankton productivity in the lower Sulphur River (Texas-Arkansas) to bleach-kraft mill effluent (BKME) were monitored using in situ ¹⁴C incubation. Carbon assimilation rates measured downstream of mill discharge were substantially reduced from upstream levels. Periphyton and phytoplankton chlorophyll a concentrations remained relatively unchanged by the presence of BKME. Periphyton ash-free dry weight increased near the mill outfall, but decreased further downstream. Calculated productivity efficiencies (productivity:biomass) varied with variations in ¹⁴C rates. A laboratory bioassay was designed to determine the effect of BKME light-attenuation on photosynthetic rates of upstream Sulphur River periphyton and Selenastrum capricornutum Prinz. Pooled results of bioassay runs indicated a 20 per cent BKME concentration effectively reduced control ¹⁴C-assimilation levels by 50 per cent. The downstream reduction observed for in situ productivity was 5 per cent lower than that predicted by the color bioassay.

It was found that Azotobacter vinelandii grown in a dialyzed soil medium did not fix nitrogen and had a much lower rate of respiration than when grown in Burk's nitrogen-free medium. When para-hydroxybenzoic acid served as the added oxidizable organic carbon source in dialyzed soil medium, the azotobacter grown in it were found to be unable to fix nitrogen. On the other hand, A. vinelandii fixed nitrogen when grown in soil supplemented with glucose. It was concluded that natural conditions in the soil are not conducive to nitrogen fixation by A. vinelandii.

Aldose reductase inhibitors (ARI's) have been shown to attenuate or prevent several complications of diabetes in animals. Tolrestat is a potent and unique ARI from Ayerst Laboratories, New York, NY. The efficacy and toxicology of tolrestat via topical ocular administration was examined. Tolrestat effectively enhanced corneal reepithelialization and was efficacious in the prevention of cataracts in the streptozotocin-diabetic rat. Ocular tissues were examined by slit lamp biomicroscopy, scanning and transmission electron microscopy, and light microscopy. Liver and kidney tissue was also examined. The presence of tolrestat was confirmed in urine, feces, and eye specimens, and quantitated in serum. There was no evidence of local or systemic tolrestat induced toxicity. Tolrestat prevented cataract formation at less than one-third the reported oral dose and at approximately one-fiftieth the associated serum concentration in rats. ED-50 and TD-50 calculations indicate that tolrestat is a relatively safe drug by topical ocular administration.

A new rapid computer assisted micro-titer plate interferon assay system was developed and characterized for use in high capacity clinical and research applications. The biological aspect of the assay was a modification of the assay methods of Finter, Armstrong and McManus. It was an application of spectrophotometric quantification of the reduction of viral cytopathic effect (CPE) as reflected by neutral red dye uptake by viable cells. A computer program was developed for the extrapolation of raw data to reference interferon units.

Bluegill were exposed to 3 and 30 pg/L lindane and 20 and 200 pg/L naphthalene to determine uptake rate constants, K1 depuration rate constants, K2, and bioconcentration factors, BCF. Correlations were determined between lipid normalized and non-lipid normalized BCFs, and between observed Kl, K2 and BCFs and predicted values. The K1 values for both chemicals and concentrations were similar. The K2 values were different (1.04 day~1, 0.46 day 1). Naphthalene was more rapid. BCFs for lindane (315) and naphthalene (98) were different. Lipid normalized BCFs for naphthalene were more variable than non-lipid normalized BCFs. The reverse was observed for lindane BCFs. Predicted K1, K2 , and BCFs were in agreement with observed values.

Biotransformation and biodegradation rate constants were determined for naphthalene, lindane, and phenol in water samples from three different sources. Rate constants produced from monitoring disappearance of the parent chemical (biotransformation) were compared to those obtained from mineralization of the chemical (ultimate biodegradation) by ¹⁴CO₂ evolution as well as acidification of the residual ¹⁴C-labeled compound (primary biodegradation). Rate constants were statistically different for the three chemicals. The water source affected the rate constants. When biomass measurements of the waters were considered and second-order rate constants were derived, there was no statistical evidence that this parameter gave a reliable rate constant statistic that could be useful in predicting the fate of any of naphthalene, lindane, and phenol in these waters.

The effect of ethanol on the stress response in rats was examined. Experimental animals were given 0.25 ml of 28 percent ethanol or 0.25 ml of water orally once a day, five days a week, for a period of twelve months and were then subjected to fifteen minute cold stress. Corticosterone levels in ethanol-treated males following stress were significantly lower (22 percent) than in the sham group. Adrenal weights in sham-treated females were significantly higher (15 percent) than in the ethanol group at the end of twelve months. Mortality in sham-treated males was significantly higher (60 percent) than in ethanol-treated males. The effects observed may be due to the sedative action of ethanol on cortical centers controlling the hypothalmus.