Simultaneous determination of uric acid metabolites allantoin, 6-aminouracil, and triuret in human urine using liquid chromatography-mass spectrometry.
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Upstream induction sequence, the cis-acting element required for response to the allantoin pathway inducer and enhancement of operation of the nitrogen-regulated upstream activation sequence in Saccharomyces cerevisiae.
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Expression of the DAL2, DAL4, DAL7, DUR1,2, and DUR3 genes in Saccharomyces cerevisiae is induced by the presence of allophanate, the last intermediate of the allantoin degradative pathway. Analysis of the DAL7 5'-flanking region identified an element, designated the DAL upstream induction sequence (DAL UIS), required for response to inducer. The operation of this cis-acting element requires functional DAL81 and DAL82 gene products. We determined the DAL UIS structure by using saturation mutagenesis. A specific dodecanucleotide sequence is the minimum required for response of reporter gene transcription to inducer. There are two copies of the sequence in the 5'-flanking region of the DAL7 gene. There are one or more copies of the sequence upstream of each allantoin pathway gene that responds to inducer. The sequence is also found 5' of the allophanate-inducible CAR2 gene as well. No such sequences were detected upstream of allantoin pathway genes that do not respond to the presence of inducer. We also demonstrated that the presence of a UIS element adjacent to the nitrogen-regulated upstream activation sequence significantly enhances its operation. (+info)
Allantoin transport in Saccharomyces cerevisiae.
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Allantoin uptake in both growing and resting cultures of Saccharomyces cerevisiae occurs by a low-Km (ca. 15 micrometer) transport system that uses energy that is likely generated in the cytoplasm. This conclusion was based on the observation that transport did not occur in the absence of glucose or the presence of dinitrophenol, carbonyl cyanide-m-chloro-phenyl hydrazine, fluoride, or arsenate ions. Normal uptake was observed, however, in the presence of cyanide. The rate of accumulation was maximal at pH 5.2. In contrast to the urea transport system, allantoin uptake appeared to be unidirectional. Preloaded, radioactive allantoin was not lost from cells suspended in allantoin-free buffer and did not exchange with exogenously added, nonradioactive allantoin. Treatment of preloaded cells with nystatin, however, released the accumulated radioactivity. Allantoin accumulated within cells was isolated and shown to be chemically unaltered. (+info)
Expression of the DAL80 gene, whose product is homologous to the GATA factors and is a negative regulator of multiple nitrogen catabolic genes in Saccharomyces cerevisiae, is sensitive to nitrogen catabolite repression.
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We have cloned the negative regulatory gene (DAL80) of the allantoin catabolic pathway, characterized its structure, and determined the physiological conditions that control DAL80 expression and its influence on the expression of nitrogen catabolic genes. Disruption of the DAL80 gene demonstrated that it regulates multiple nitrogen catabolic pathways. Inducer-independent expression was observed for the allantoin pathway genes DAL7 and DUR1,2, as well as the UGA1 gene required for gamma-aminobutyrate catabolism in the disruption mutant. DAL80 transcription was itself highly sensitive to nitrogen catabolite repression (NCR), and its promoter contained 12 sequences homologous to the NCR-sensitive UASNTR. The deduced DAL80 protein structure contains zinc finger and coiled-coil motifs. The DAL80 zinc finger motif possessed high homology to the transcriptional activator proteins required for expression of NCR-sensitive genes in fungi and the yeast GLN3 gene product required for functioning of the NCR-sensitive DAL UASNTR. It was also homologous to the three GATAA-binding proteins reported to be transcriptional activators in avian and mammalian tissues. The latter correlations raise the possibility that both positive and negative regulators of allantoin pathway transcription may bind to similar sequences. (+info)
Absolute configurations of spiroiminodihydantoin and allantoin stereoisomers: comparison of computed and measured electronic circular dichroism spectra.
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Effect of cepea extract-heparin and allantoin mixture on epidural fibrosis in a rat hemilaminectomy model.
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AIM: Epidural fibrosis following a laminectomy procedure is a serious problem that results in failed back surgery syndrome. Aserious number of manuscripts have explained its possible mechanism and results but no effective preventive surgical technique or treatment is currently present. MATERIAL AND METHODS: We used a rat hemilaminectomy model at lumbar fourth level. In the treatment group (n:10), the hemilaminectomy sites were filled with cepea extract-allantoin and heparin mixture as sterile cream form. In the second group, the same surgical procedure was performed and the site was filled with physiological saline. All animals were terminated after 6 weeks and laminectomy sites removed en-bloc. Epidural fibrosis was evaluated and compared using semi-quantitative histopathological scoring scales. RESULTS: In the physiological saline group, the fibrosis score was 10.3 points and 90% of the subjects had acute inflammatory reaction, 80% chronic inflammatory reaction and 100% showed bone destruction and reparation process. In the cepea extract group, these values were fibrosis score 4.2 points, 0% acute inflammatory reaction, 33.3% chronic inflammatory reaction and 10% bone destruction and reparation process, respectively. CONCLUSION: This study showed that aloe cepea extract-allantoin and heparin mixture diminished epidural scarring formation effectively with decreased scores of acute and chronic inflammation, compared to the physiological saline solution group. (+info)
Nitrogen utilization in growing lambs: effects of grain (starch) and protein sources with various rates of ruminal degradation.
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The potential interaction between grain (starch) and protein sources with varying ruminal degradation rates on N utilization in growing lambs was evaluated. Three grain sources with varying ruminal degradation rates, (barley greater than steam-flaked sorghum [SFSG] greater than dry-rolled sorghum [DRSG]) and three protein sources (urea greater than a 50:25:25 mixture of urea: blood meal:corn gluten meal [N basis, U/BC] greater than 50:50 mixture of meal:corn gluten meal [N basis, BC]), were evaluated in a 3 x 3 factorial arrangement. Supplemental protein sources provided 33% of dietary N (CP = 11.0%). For each grain-protein combination, a 3 x 3 Latin square metabolism trial was conducted using two sets of three lambs and three periods. Within-square treatments were 1.4, 1.7 and 2.0 times maintenance intake levels. No interactions were observed (P greater than .2) between dietary treatments and intake level. Grain sources did not differ (P greater than .2) in N balance or the proportion of N retained. Lambs fed urea diets retained less N (3.6 vs 4.2 and 4.1 g/d for urea vs U/BC and BC, respectively; linear, P = .07; quadratic, P = .12) and utilized N less efficiently (43.1 vs 51.9 and 52.5%, respectively; linear, P less than .001; quadratic, P = .10) than lambs fed BC diets. The grain x protein interaction was significant for most variables. Nitrogen utilization was most efficient (24 to 27% of N intake retained) when rapidly degraded sources (barley and urea) and slowly degraded sources (sorghum and BC) were fed together or when U/BC was the supplemental protein source (interaction P less than .08). An advantage was found for selection of starch and protein sources with similar ruminal degradation rates. (+info)
Renal clearance of plasma allantoin in sheep.
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The recovery in urine of an intrajugular infusion of physiological amounts of allantoin was measured in four sheep nourished by an intragastric infusion of volatile fatty acids and casein (to eliminate rumen fermentation). The recovery was 72% (S.E.M. 7) and the remainder was presumed to have been lost by diffusion into the gut and degradation by gut microflora. Measured in two sheep, allantoin was removed from the blood at a fractional rate of 0.30 h-1, and excreted in urine at 0.23 h-1. Calculation based on creatinine excretion showed glomerular filtration rate and tubular reabsorption of allantoin to be unchanged by the intravenous infusion. Maximal tubular reabsorption at 1.28 mmol day-1 was saturated by the load of endogenous allantoin alone. In a second experiment with seven normally fed sheep (28-50 kg live weight, all given 1 kg feed), urinary excretion and plasma concentration of allantoin were linearly related. However, the errors were such that plasma allantoin concentration would be of little value as a predictor of urinary excretion. There was a nearly twofold range in allantoin excretion (the larger animals excreting less), which implied that the supply of microbial biomass to the host animal per unit of feed ingested could be profoundly affected by feeding level. (+info)