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Quantitation of 5-methylcytosine in DNA after acid hydrolysis has been inaccurate because deamination of cytosine and 5-methylcytosine occurs during the hydrolysis procedure. There is little information in the literature regarding the use of hydrofluoric acid (HF) for DNA hydrolysis and we have therefore undertaken a systematic study of this process. The deoxyribonucleotides of cytosine and 5-methylcytosine were shown not to undergo detectable levels of deamination during prolonged periods (up to 24 h) at 80 degrees C in 48% HF. Kinetic studies show that the release of purine and pyrimidine bases was complete by 4 h under these conditions. Analysis of the 5-methylcytosine content of DNA from various tissues gave levels that were very close to the values reported in the literature. This method is ideally suited for the determination of the overall cytosine methylation levels in DNA. 相似文献
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Michael V. Keenan William L. Alworth 《Biochemical and biophysical research communications》1974,57(2):500-504
Δ-1-Pyrroline-2-carboxylate is a potent inhibitor of proline racemase from . It is proposed that Δ-1-pyrroline-2-carboxylate acts as an analogue of the transition state of the catalyzed reaction. 4,5-Imidazoledicarboxylate, 2,3-pyridinedicarboxylate, and 2-benzimidazolecarboxylate, when present at concentrations equal to that of the substrate, -proline, failed to inhibit proline racemase. 相似文献
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Male Fischer 344 rats classified as young (2-4 months), middle-aged (12-14 months) and aged (22-25 months) received 300, 600 or 800 mg/kg acetaminophen (APAP) intraperitoneally and were sacrificed 24 hr later. Blood urea nitrogen (BUN) concentration and urinary glucose and osmolality were determined. In addition, kidneys were evaluated for histopathological changes. APAP did not affect osmolality or BUN concentrations and failed to produce lesions after any dose in young rats. Osmolality was decreased 40% and 50% in middle-aged and aged rats, respectively, after 800 mg/kg APAP. Glucosuria was prominent in aged rats after the 600 and 800 mg/kg doses were administered, while middle-aged rats showed little glucosuria after these doses. BUN concentrations were elevated 89% and 183% in middle-aged and aged rats, respectively, given 600 mg/kg APAP; after 800 mg/kg, BUN concentrations were elevated approximately four-fold in both age groups. Pathological evaluations showed a greater incidence of acute tubular necrosis (ATN) in aged kidneys compared to kidneys of middle-aged rats after 600 mg/kg, while the two older groups exhibited similar, more severe ATN after 800 mg/kg APAP. These data suggest an age-related increased susceptibility of male Fisher 344 rats to APAP nephrotoxicity. 相似文献