Microbial biogeochemistry of uranium mill tailings.
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Whereas in freely suspended cell cultures growing photoautotrophically under non-limiting carbon conditions nitrite and nitrate were simultaneously consumed after ammonium consumption was complete, in alginate-entrapped cell cultures a sequential consumption of nitrite (first) and nitrate was observed after ammonium had almost been fully removed. In this paper results are reported that show inhibition of nitrate consumption by nitrite in immobilized cells. However no inhibition of nitrate active transport was observed. The sequential consumption of ammonium, nitrite and nitrate by Ca-alginate immobilized cells is explained on the basis of local ammonium accumulation due to its photoproduction by photorespiration, that could be caused by the increase of the O2/CO2 ratio around the entrapped cells. Measurements of light-dependent oxygen production (LDOP) and activity levels of nitrogen assimilation enzymes, including nitrite reductase (NiR) and glutamine synthetase (GS) in immobilized cells, determined under photorespiration stimulating conditions, are shown that support this explanation.
The effect of 60 minutes' intravenous infusions, before morning feeding, of ammonium acetate (18.6 micromole/min/kg of body weight) and ammonium acetate with propranolol (11 microgram/min/kg of body weight) on the ruminal motility of sheep was examined. Ammonia has a adrenaline-like action therefore propranolol, a beta-receptor blocking agent, was administered in order to eliminate the possible effect of adrenaline on ruminal motility. The contractions of the dorsal sack of the rumen were registered by means of the balloon method, with the gauge inserted through the rumen fistula. The infusion of ammonium acetate caused an increase of the ammonia concentration in the blood to 0.6 mmol/1 at the end of 60 minutes' infusion. Already during the first 5 minutes of the intravenous infusion of ammonium acetate there was a decreased frequency of ruminal contractions, which was observed throughout the infusion. After the infusion there was a radical decrease of the concentration of ammonia in the blood, and at the same time an increase in the frequency of rumen contraction was observed. Blocking of the beta-adrenergic receptors by propranolol did not eliminate the inhibiting action of ammonium ion on rumen motility. The infusion of the ammonium acetate caused an increase of adrenaline and glucose concentration. This response was eliminated by propranolol in the case of adrenaline but not glucose. It is assumed that the action of ammonium ion on the rumen motility is derived primarily by the central nervous system.
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The activities of citrate synthase (EC 4.1.3.7) and NADP+-dependent glutamate dehydrogenase (GDH) (EC 1.4.1.4) of Saccharomyces cerevisiae were inhibited in vitro by glyoxylate. In the presence of glyoxylate, pyruvate and glyoxylate pools increased, suggesting that glyoxylate was efficiently transported and catabolized. Pyruvate accumulation also indicates that citrate synthase was inhibited. A decrease in the glutamate pool was also observed under these conditions. This can be attributed to an increased transamination rate and to the inhibitory effect of glyoxylate on NADP+-dependent GDH. Furthermore, the increase in the ammonium pool in the presence of glyoxylate suggests that NADP+-dependent GDH was being inhibited in vivo, since the activity of glutamine synthetase did not decrease under these conditions. We propose that the inhibition of both citrate synthase and NADP+-dependent GDH could form part of a mechanism that regulates the internal 2-oxoglutarate concentration.
As measures are implemented internationally to reduce SO2 and NOx emissions, attention is falling on the contribution of NH3 emissions to acidification, nitrogen eutrophication, and aerosol formation. In the U.K., a monitoring network has been established to measure the spatial distribution and long-term trends in atmospheric gaseous NH3 and aerosol NH4+. At the same time, an atmospheric chemistry and transport model, FRAME, has been developed with a focus on reduced nitrogen (NHx). The monitoring data are important to evaluate the model, while the model is essential for a more detailed spatial assessment. The national network is established with over 80 sampling locations. Measurements of NH3 and NH4+ (at up to 50 sites) have been made using a new low-cost denuder-filterpack system. Additionally, improved passive sampling methods for NH3 have been applied to explore local variability. The measurements confirm the high spatial variability of NH3 (annual means 0.06 to 11 microg NH3 m(-3)), consistent with its nature as a primary pollutant emitted from ground-level sources, while NH4+, being a slowly formed secondary product, shows much less spatial variability (0.14 to 2.4 mg NH4+ m(-3)). These features are reproduced in the FRAME model, which provides estimates at a 5-km level. Analysis of the underlying NH3 emission inventory shows that sheep emissions may have been underestimated and nonagricultural sources overestimated relative to emissions from cattle. The combination of model and measurements is applied to estimate spatial patterns of dry deposition to different vegetation types. The combined approach provides the basis to assess NHx responses across the U.K. to international emission controls.
Rats were fed the following diets: standard (20% protein), high-protein (80%), protein-free, standard plus ammonium and protein-free plus ammonium for six weeks. The standard plus ammonium diet was prepared to contain ammonia equivalent to that supplied by the high-protein diet. Addition of ammonium acetate (20% by mass) to the 20% protein or protein-free diets results in 2.3- and 10-fold increases of urea excretion respectively, without increase of carbamoyl-phosphate synthase. Supplementation of the standard diet with ammonium increases the mitochondrial content of acetylglutamate from 830 to 1590 pmol/mg protein, and of the protein-free diet from 130 to 1040 pmol/mg. However, ingestion of ammonium did not increase the activity of acetylglutamate synthase. Therefore the efflux of acetylglutamate from mitochondria was determined. After 30 min at 37 degrees C liver mitochondria from rats on standard diet released 61% of the initial acetylglutamate while mitochondria from animals on standard plus ammonium diet released only 20%. These results indicate that ingestion of ammonium increases the content of acetylglutamate in rat liver by decreasing its efflux from mitochondria. This effect is similar to that produced in mice by a high protein diet [Morita et al. (1982) J. Biochem. (Tokyo) 91, 563-569]. However, while the high-protein diet increases carbamoylphosphate synthase content, the ammonium diet does not.
Ingestion of large amounts of ammonium increases markedly the content of tubulin in brain. The effect on tubulin induction of ammonium ingestion for up to 100 days was investigated. Brain tubulin content showed a rapid initial increase (28%) at 2 days and reached 50% after 100 days on the diet. To discern if ammonia, the increase in urea synthesis, or both was responsible for tubulin induction, rats were maintained at several levels of uremia (by administering diets containing 0 to 80% protein) or in hyperammonemia (by urease treatment). Only ammonium administration in the diet and urease injection induced tubulin in brain. Tubulin was quantified in three different brain regions. There was a regional selectivity of tubulin induction by ammonia in rat brain. Whereas the cerebellum remained unaltered, the paleencephalon showed the highest increase, and the cerebral cortex exhibited only a modest increase.
The two glutamine synthetases of Rhizobium sp. 32H1 appear to be structurally and functionally distinct. Glutamine synthetase I was reversibly adenylylated, and its synthesis was repressed only twofold by ammonium. When in the unadenylylated configuration, it was the enzyme which allowed the organism to grow, albeit marginally, on ammonium as a nitrogen source. There is no evidence to suggest that the second enzyme, glutamine synthetase II, is regulated by adenylylation. However, this enzyme was repressed at least 50-fold by even low amounts of ammonium. Glutamine synthetase II does not seem to function in ammonium assimilation but rather in purine biosynthesis.
The effect of L-methionine-DL-sulfoximine, an inhibitor of glutamine synthetase, on the formation of nitrate reductase in the wild-type strain of Neurospora in the presence of ammonium ions and of glutamine was studied. Under conditions in which glutamine synthetase was inactivated, it was found that only glutamine could repress nitrate reductase. In a mutant of Neurospora, gln-1b, which requires glutamine for growth, only glutamine could repress nitrate reductase. These results suggest a direct role for glutamine as corepressor of nitrate reductase in Neurospora.
Rhizobium phaseoli lacks glutamate dehydrogenase (GDH) and assimilates ammonium by the glutamine synthetase-glutamate synthase pathway. A strain of R. phaseoli harboring the Escherichia coli GDH structural gene (gdhA) was constructed. GDH activity was expressed in R. phaseoli in the free-living state and in symbiosis. Nodules with bacteroids that expressed GDH activity had severe impairment of nitrogen fixation. Also, R. phaseoli cells that lost GDH activity and assimilated ammonium by the glutamine synthetase-glutamate synthase pathway preferentially nodulated Phaseolus vulgaris.
T-strain mycoplasmas require urea for propagation, but urea metabolism also occurs in nonpropagating viable cultures. Ammonia results from this metabolism and alkalinizes the medium. Ammonium ions and an alkaline pH both inhibit the multiplication of T strains and reduce the viability of T strains in broth. These toxic effects of urea metabolism currently limit the growth of T strains in broth. Stock T-strain cultures are optimally maintained in continuous culture if the routine medium at pH 6.0 is supplemented with 0.05% urea and 0.002% phenol red, but an incubation temperature of 30 C is preferable to 37 C for subculture at 24-hr intervals.
Glutamine synthetases (GS) are key enzymes of nitrogen metabolism. Most bacteria contain only one type of GS enzyme encoded by glnA. Streptomyces coelicolor, the model organism for Gram-positive streptomycetes, however is characterized by two functional GS (glnA, glnII) involved in nitrogen assimilation. In addition, three GS-like genes were identified which do not exhibit GS enzyme activity. The control of nitrogen assimilation and metabolism is mediated by transcriptional and post-translational regulation systems. The OmpR-like regulators GlnR and GlnRII are involved in transcriptional control of important nitrogen metabolism genes (glnA, glnII, amtB, glnK, glnD). Although GlnR and GlnRII share identical binding regions, their physiological impact is different. GSI activity is modulated post-translationally by the adenylyltransferase GlnE in response to the nitrogen concentration whereas no post-translational modifications of GSII are known. The PII/GlnD system also responds to changes in nitrogen conditions. The adenylyltransferase GlnD, which resembles the uridylyltransferase of Enterobacteriaceae, modifies PII under low-nitrogen conditions. Furthermore, PII is processed at its N-terminus in response to an ammonium shock. Apparently the function of the PII protein of S. coelicolor is different from that of the PII proteins of Enterobacteriaceae.
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Basidia of Coprinus cinereus (Schaeff.:Fr) S. F. Gray are committed to their developmental pathway, continuing through meiosis and sporulation even when excised from their parental fruit body. A technique is described which permits this in vitro differentiation to be used as a rapid, small-scale bioassay for chemicals which interfere with these morphogenetic processes. Of a range of compounds tested, only ammonium and glutamine, and some structural analogues, were able to inhibit basidium differentiation. Growth was not inhibited; instead the differentiation inhibitors caused vegetative hyphal tips to grow out from regions of the basidial apparatus expected to be in active growth during sporulation. Depending on the stage reached at the time of exposure to the inhibitors, vegetative hyphal tips emerged from the four apical sites for sterigmata, from the tips of sterigmata, from partially formed or abnormal spores, and from the basal regions of the basidium from which paraphyses would be expected to arise. The experiments show that ammonium ions and glutamine halt meiocyte differentiation. Reports of similar effects in other organisms, animals and plants as well as fungi, may imply that sporulation events are generally sensitive to ammonium inhibition.
Investigations of the synthesis of organic compounds from methane and ammonia were carried out by using high frequency discharges. The radical of CN was observed in the early phase of the discharge. Therefore it is considered that the CN radical contributes to the successive reaction steps. An important intermediate, a tetramer of hydrogen cyanide was observed to be formed at room temperature. The amounts of this compound progressively increased with time. It was speculated that the purines which were identified in the refluxed sample were synthesized through the above tetramer. Compounds which are thought to be heterocyclic (max:250-265nm) were initially formed in the sample trapped at liquid nitrogen temperature. After hydrolysis with hydrochloric acid, Gly, Asp Ala, etc. were observed in the sample. Overall yield of organic compounds was estimates at 5-6% based on input carbon of methane.