Synthesis and degradation of xanthine dehydrogenase in chick liver. In vivo and in vitro studies.

The present study describes the (xanthine:NAD+ oxidoreductase, EC 1.2.1.37) synthesis and degradation of chick liver xanthine dehydrogenase in vivo and in organ cultures. The results indicate that control of xanthine dehydrogenase activity is mediated by changes in the rate of enzyme synthesis, but that degradation rates are unaffected. The results also suggest that xanthine dehydrogenase synthesis occurs through a previously unreported intermediate. Detected in cultures of liver tissue, this intermediate apparently is not converted into an active enzyme. A model of synthesis and degradation for xanthine dehydrogenase proposes that the synthesis of the enzyme is proportional to messenger RNA and includes an inactive enzyme precursor and a second inactive intermediate prior to degradation. Integrated mathematical solutions describing the concentration of intermediates as a function of time can be found explicitly for simple models. The appendix to this paper extrapolates solutions for one-, two- and three-step models to generate a mathematical solution for an 'n'-step model containing 'n' intermediates. The rate constants in the solutions can be found experimentally.

Determination of the sedimentary microbial biomass by extractible lipid phosphate.

The measurement of lipid phosphate is proposed as an indicator of microbial biomass in marine and estuarine sediments. This relatively simple assay can be performed on fresh, frozen or frozen-lyophilized sediment samples with chloroform methanol extraction and subsequent phosphate determination. The sedimentary lipid phosphate recovery correlates with the extractible ATP and the rate of DNA synthesis. Pulse-chase experiments show active metabolism of the sedimentary phospholipids. The recovery of added 14C-labeled bacterial lipids from sediments is quantitative. Replicate analyses from a single sediment sample gave a standard deviation of 11%. The lipid extract can be fractionated by relatively simple procedures and the plasmalogen, diacyl phospholipid, phosphonolipid and non-hydrolyzable phospholipid content determined. The relative fatty acid composition can be readily determined by gas-liquid chromatography.The lipid composition can be used to define the microbial community structure. For example, the absence of polyenoic fatty acids indicates minimal contamination with benthic micro-eukaryotes. Therefore the high content of plasmalogen phospholipids in these sediments suggests that the anaerobic prokaryotic Clostridia are found in the aerobic sedimentary horizon. This would require anaerobic microhabitats in the aerated zones.

Poly-beta-Hydroxybutyrate Accumulation as a Measure of Unbalanced Growth of the Estuarine Detrital Microbiota.

The procaryotic endogenous storage material poly-beta-hydroxybutyrate (PHB) can be induced to accumulate in the estuarine detrital microbiota under conditions which suggest unbalanced growth, such as limitation of a critical factor(s) in the presence of carbon and energy sources. Changes in PHB-to-lipid phosphate ratios detected in field samples can be mimicked in the laboratory with common estuarine stresses. Acute anoxia or low pH induces conditions of no growth with depression of both the synthesis and catabolism of PHB without change in the lipid phosphate. Balanced growth induced by nutrients increases the lipid phosphate, depresses PHB synthesis, and stimulates PHB catabolism, resulting in a low ratio of PHB to lipid phosphate. Unbalanced growth induced to a small extent by high salinity or much more readily by dark upland runoff water results in rapid accumulation of PHB and slowing of PHB catabolism with little change in lipid phospate. Unbalanced growth conditions result in high PHB-to-lipid phosphate ratios in the detrital microbiota.

Regulation of nitrogen catabolic enzymes in chick liver: effects of insulin.

Previous studies have shown that a group of nitrogen catabolic enzymes including xanthine dehydrogenase, purine nucleoside phosphorylase, and tyrosine aminotransferase are all increased in chick liver by dietary protein as well as single amino acids (e.g. methionine) and certain antimetabolites (e.g. hydrazine). A similar enzyme response pattern can be obtained with insulin. This hormone causes an enhanced rate of XDH synthesis and gives nonadditive results with protein, hydrazine and methionine. Furthermore, a vitamin B6 dependency was observed in responses to both high protein diets and insulin, all suggesting a common regulatory mechanism. In this system dietary protein and insulin may act similarly by increasing the availability of amino acids to the liver -- in one case by supplying amino acids through the diet and in the other by increasing amino acid uptake.

Variation in Phospholipid Ester-Linked Fatty Acids and Carotenoids of Desiccated Nostoc commune (Cyanobacteria) from Different Geographic Locations.

Profiles of phospholipid fatty acids and carotenoids in desiccated Nostoc commune (cyanobacteria) collected from China, Federal Republic of Germany, and Antarctica and in axenic cultures of the desiccation-tolerant strains N. commune UTEX 584 and Hydrocoleum strain GOEI were analyzed. The phospholipid fatty acid contents of the three samples of desiccated Nostoc species were all similar, and the dominant compounds were 16:1omega7c, 16:0, 18:2omega6, 18:3omega3, and 18:1omega7c. In comparison with the field materials, N. commune UTEX 584 had a much higher ratio of 18:2omega6 to 18:3omega3 (5.36) and a significantly lower ratio of 18:1omega7c to 18:1omega9c (1.86). Compound 18:3 was present in large amounts in the samples of desiccated Nostoc species which had been subject, in situ, to repeated cycles of drying and rewetting, but represented only a small fraction of the total fatty acids of the strains grown in liquid culture. This finding is in contrast to the data obtained from studies on the effects of drought and water stress on higher plants. Field materials of Nostoc species contained, in contrast to the axenic strains, significant amounts of apocarotenoids and a P384 pigment which, upon reduction with NaBH(4), yielded a mixture of a chlorophyll derivative and a compound with an absorption maximum of 451 nm. A clear distinction can be made between the carotenoid contents of the axenic cultures and the desiccated field materials. In the former, beta-carotene and echinenone predominate; in the latter, canthaxanthin and the beta-gamma series of carotenoids are found.

Effects of oil and gas well-drilling fluids on the biomass and community structure of microbiota that colonize sands in running seawater.

Well-drilling fluid and a number of the known components (barite, clay, Aldacide, Surflo, and Dowicide, were tested for effects on the biomass and community structure of the microbiota that colonize marine sands exposed for eight weeks to running ambient seawater. Shading the microbiota from light depressed the microflora without a significant effect on the biomass, while well-drilling fluids layered on the surface or mixed with the sand significantly increased a component of the bacteria and the microfauna as reflected in changes in the fatty acid composition. There were some shading effects from the surface layering of well-drilling fluids as reflected in the fatty acids from the microflora when compared to the sands mixed with well-drilling fluids. Barite had essentially no effect on the biomass or community structure while clays increased nearly all of the biomass indicators for the bacteria as well as the microfauna; the clay overlay mirrors the effect of the drilling fluids. Aldacide shifted the bacterial composition, depressing the proportions of microbes containing the cyclopropane fatty acids and the anaerobic pathways of desaturation. Concentrations of 1 and 15 microgram/L increased the bacterial biomass as reflected in the total lipid (16:0) and extractable lipid phosphate coupled with a decrease in the total microeukaryotes. Surflo increased the biomass and shifted the bacterial community structure at concentrations between 4 and 800 microgram/L. The lowest level also stimulated the microfauna. Dowicide at 100 microgram/L increased the bacteria forming cis-vaccenic acid and the microfauna similar to low concentrations of Surflo.

Effect of silicate grain shape, structure, and location on the biomass and community structure of colonizing marine microbiota.

Microbiota colonizing silica grains of the same size and water pore space, but with a different microtopography, showed differences in biomass and community structure after 8 weeks of exposure to running seawater. The absence of surface cracks and crevices resulted in a marked diminution of the total microbial biomass measured as lipid phosphate and total extractable palmitic acid. With increasing smoothness of the sand grain surface, examination of the community structure showed a marked decrease in procaryotes and algal microeucaryotes, with a relative increase in microeucaryotic grazers. A comparison of the colonizing sediment incubated in running seawater or at 32 m on the sea floor with a sediment core showed a decreased bacterial biomass with a different community structure and a decreased total microeucaryotic population of both grazers and algae. The quantitative differences in microbial biomass and community structure between the microcosms and the actual benthic population in the core were determined.

Effect of manual brush cleaning on biomass and community structure of microfouling film formed on aluminum and titanium surfaces exposed to rapidly flowing seawater.

Metals exposed to rapidly flowing seawater are fouled by microbes that increase heat transfer resistance. In this study, results of biochemical test methods quantitatively relating the biomass and community structure of the microfouling film on aluminum and titanium to heat transfer resistance across the metal surface during three cycles of free fouling and manual brushing showed that cleaning accelerates the rate of fouling measured as the loss of heat transfer efficiency and as microfouling film biomass. The results also showed that the rate of fouling, measured as an increase in heat transfer resistance, is faster on titanium than on aluminum but that the titanium surface is more readily cleaned. In three cycles of free fouling and cleaning with a stiff-bristle nylon brush, the free-fouling communities re-forming on aluminum became enriched in bacteria containing short-branched fatty acids as the cycling progressed. The free-fouling community on titanium revealed an increasingly diverse morphology under scanning electron microscopy that was enriched in a portion of the microeucaryotes. Brushing removed most of the biomass, but left a residual community that was relatively enriched in a portion of the bacterial assembly containing cyclopropane fatty acids on aluminum and in a more diverse community on the titanium surface. The residual communities left after cleaning of titanium revealed an increase in bacteria with short-branched fatty acids and in microeucaryotes as cleaning continued. No significant changes occurred in the residual microbial community structure left on aluminum with cleaning; it was, again, less diverse than that remaining on titanium. The residual communities secreted a twofold-larger amount of extracellular polymer, measured as the ratio of total organic carbon to lipid phosphate, than did the free-fouling community on both surfaces.

Regulation of nitrogen catabolic enzymes in chick liver -- effects of amino acids.

It is well-known that in the chick, dietary protein increases the levels of several hepatic enzymes that are involved in nitrogen metabolism and excretion. However, the biochemical mechanism of this response is essentially unknown. The experiments presented in this paper show that the chick is responding to alpha-amino nitrogen and not to any specific amino acid. Furthermore, it is shown that this system responds to endogenous sources of nitrogen as well as dietary protein and that the xanthine dehydrogenase response involves regulation of enzyme synthesis without changing the rate of degradation.

Effect of light on biomass and community structure of estuarine detrital microbiota.

Comparison of estuarine detrital microbiota grown with and without light in the absence of macroscopic grazing showed shifts in the community structure that enabled correlation between various biochemical measures. Analysis of these biochemical measures showed that growth in light induces the smallest increases in procaryotic attributes such as muramic acid; wall glucosamine; lipid phosphate; total extractable adenosine nucleotides; short-branched, cyclopropane, and cis-vaccenic fatty acids; lipid glucose and mannose; the incorporation of acetate into lipid; and the formation of deoxyribonucleic acid from thymidine. Measures of the microfauna such as lipid inositol and the gamma-linolenic series of polyenoic fatty acids also increased minimally in the light-grown microbiota. Measures of sulfo-lipid synthesis, lipid glycerol, total extractable palmitate, 18-carbon polyenoic fatty acids, and total polyenoic fatty acids longer than 20 carbons increased 10- to 15-fold in algae and fungi. Chlorophyll a, lipid galactose, and the 16- and 20- carbon polyenoic fatty acids characteristic of diatoms increased maximally in the light. This increase of diatom measure correlated with the sheets of diatoms detected by scanning electron microscopy.