Dihydrofolate reductase from trimethoprim-resistant Escherichia coli MB 3746 and MB 3747. Purification, amino acid composition, and some kinetic properties.
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Biomedical subjects
Publications and source records attributed to C R Short.
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The retention of 3-methylcholanthrene (3-MC) in rat hepatic cytosol was significantly enhanced by adrenalectomy. In contrast, there was no significant difference in 3-MC retention in females as compared with males. 3-MC present in the cytosol fraction was bound to macromolecules and could be separated into three fractions by ion-exchange column chromatography.
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A simple, rapid extraction and subsequent determination for strychnine, using high pressure liquid chromatography (HPLC) with a reverse phase solvent system, is described. Stomach contents or grain bait containing strychnine were made alkaline with sodium hydroxide and extracted with chloroform. Extract filtrates were injected directly into a liquid chromatograph without further preparation except for dilution, if necessary. Peaks were resolved within 3.5 min and peak heights were used for quantitation. HPLC of strychnine was carried out on a 30 cm X 4 mm id stainless steel column packed with micronBondapak C18. The solvent program was 0.005M phosphate buffer-methanol (60+40) at a flow of 1.5 ml/min. Recovery from spiked stomach contents was 93.9 +/- 3.5%. The detection capability for strynchnine, using the 254 nm ultraviolet detector, was 5 ng. The strychnine peak was collected and subjected to thin layer chromatography with strychnine standards for confirmation.
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The fraction binding of 14-C-salicylic acid and 14-C-pentobarbital was studied as a function of age in plasma of the pig, dog, goat and human. The pig exhibited an unusual degree of hypoalbuminemia and low fractional binding of salicylic acid at birth. Albumin levels and the percent binding of the salicylic acid were not significantly lower in newborn plasma than in adult plasma of the other species studied. The pig was also exceptional in that the high fractional binding of pentobarbital observed at low ligand concentrations in plasma of the newborn was not observed in the other species studied. Depletion of albumin in newborn pig plasma did not appreciably affect the binding of pentobarbital. Thus it is suggested that the plasma of the fetal and newborn pig may contain a nonalbuminoid protein, such as fetal globulin, which is capable of binding pentobarbital but does not have a high avidity for salicylic acid. It apparently does not exist at birth in the plasmas of the other species studied, as depletion of albumin markedly reduced the fractional binding of pentobarbital.