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Biomedical subjects

A G Clark

Publications and source records attributed to A G Clark.

230 records · Page 13Linked to original sources

Cerebral-cortex hexokinase. Elucidation of reaction mechanisms by substrate and dead-end inhibitor kinetic analysis.

1. The substrate kinetic properties of cerebral hexokinases (mitochondrial and cytoplasmic) were studied at limiting concentrations of both glucose and MgATP(2-). Primary plots of the enzymic activity gave no evidence of a Ping Pong mechanism in three types of mitochondrial preparation tested (intact and osmotically disrupted mitochondria, and the purified mitochondrial enzyme), nor in the purified cytoplasmic preparation. 2. Secondary plots of intercepts from the primary plots (1/v versus 1/s) versus reciprocal of second substrate of the mitochondrial activity gave kinetic constants which differed from those obtained directly from the plots of 1/v versus 1/s or of s/v versus s, although the ratios of the derived constants were consistent. The kinetic constants obtained with the cytoplasmic enzyme from primary and secondary plots were consistent. 3. Deoxyglucose, as alternative substrate, inhibited cytoplasmic hexokinase by competition with glucose, but did not compete when MgATP(2-) was the substrate varied. The K(i) for deoxyglucose when glucose concentrations were varied was 0.25mm. 4. A range of ATP analogues was tested as potential substrates and inhibitors of hexokinase activity. GTP, ITP, CTP, UTP and betagamma-methylene-ATP did not act as substrates, nor did they cause significant inhibition. Deoxy-ATP proved to be almost as effective a substrate as ATP. AMP inhibited but did not act as substrate. 5. N-Acetyl-glucosamine inhibited all preparations competitively when glucose was varied and non-competitively when MgATP(2-) was varied. AMP inhibition was competitive when MgATP(2-) was the substrate varied and non-competitive when glucose was varied. 6. The results are interpreted as providing evidence for a random reaction mechanism in all preparations of brain hexokinase, cytoplasmic and mitochondrial. The kinetic properties and reaction mechanism do not change on extraction and purification of the particulate enzyme. 7. The results are discussed in terms of the participation of hexokinase in regulation of cerebral glycolysis.

Adenosine Monophosphate↗

The metabolism of hexachlorocyclohexanes and pentachlorocyclohexenes in flies and grass grubs.

1. gamma-Hexachlorocyclohexane, gamma-pentachlorocyclohexene and delta-pentachlorocyclohexene were converted by houseflies and grass grubs into metabolites that had chromatographic properties identical with those of S-2,4-dichlorophenylglutathione. 2. The metabolism of gamma-hexachlorocyclohexane and the pentachlorocyclohexene isomers was negligible in newly emerged blowflies, but increased over the next 10 days. 3. The metabolism of both gamma-hexachlorocyclohexane and the pentachlorocyclohexene isomers was inhibited by simultaneous dosage with tetrabromophenolphthalein ethyl ester or Bromophenol Blue in both grass grubs and flies, but only the metabolism of pentachlorocyclohexenes in blowflies was stopped by simultaneous dosage with bis-(N-dimethylaminophenyl)methane. NN-Di-n-butyl-p-chlorobenzenesulphonamide had no effect on the metabolism of pentachlorocyclohexenes by blowflies. 4. The use of these inhibitors and colorimetric assays leads to the conclusion that a pentachlorocyclohexene is not a major intermediary metabolite of gamma-hexachlorocyclohexane in these insects.

Animals↗

Species differences in the inhibition of glutathione S-aryltransferase by phthaleins and dicarboxylic acids.

1. Glutathione-S-aryltransferase activity from grass grubs (Costelytra zealandica) was inhibited by phthaleins, sulphonphthaleins and some dicarboxylic acids. 2. These compounds had no detectable action on the enzyme from sheep liver. 3. In insect enzyme the inhibition was competitive with respect to glutathione and non-competitive with respect to the aromatic substrate. 4. Michaelis constants and inhibitor constants were measured for sheep-liver or grass-grub enzyme between pH5 and pH10 and evidence was obtained for the presence of two groups with pK9.2 in the glutathione-binding site of the insect enzyme. 5. Only one such group was detected in the sheep-liver enzyme.

Animals↗

Age-related thymic atrophy in the guinea pig.

We have characterized age-related thymic atrophy in the guinea pig, including identification of antibodies that allow immunohistochemical assessment of thymopoiesis. Age-related thymic atrophy in guinea pigs more closely resembles what occurs in humans histologically and in thymus weight, cellularity, and percent functional area than do other rodent models. The guinea pig model is thus particularly well-suited to study the role of the thymic perivascular space in age-related thymic atrophy. We next tested the hypothesis that dietary supplementation with Vitamin C could prevent or delay age-related thymic atrophy. Thymus histology, weight, cellularity, and percent functional area did not differ at 12 months between groups that received 3, 30, or 150 mg Vitamin C daily from 4 months of age. Thus long-term supplementation with up to 130 mg/kg/day Vitamin C is insufficient to influence the time course and extent of age-related thymic atrophy in guinea pigs.

Aging↗

Negotiating the transition to middle school: the role of self-regulatory processes.

The present research examined the role of maladaptive self-regulatory beliefs as vulnerability factors for academic and emotional difficulties during the transition to middle school. A short-term longitudinal design was employed to follow two groups of early adolescents: 187 adolescents who experienced a school transition between the fifth and sixth grades, and 142 adolescents who did not experience a school transition between the fifth and sixth grades. Adolescents completed measures of perceptions of academic control and importance of academic success, experience of chronic academic strain, daily school hassles, and depressive symptoms. Teachers reported on students' academic engagement, including levels of helpless behavior, effort, and academic performance. Consistent with the proposed model of self-regulation, maladaptive self-regulatory beliefs (i.e., decreased perceptions of academic control and importance) predicted individual differences in perceived school-related stress and depressive symptoms over the course of the middle school transition, but were not associated with academic and emotional difficulties in adolescents who remained in a stable school environment. Moreover, a self-regulatory sequence was identified proceeding from maladaptive self-regulatory beliefs, to academic disengagement, to enhanced perceptions of school-related stress, to depressive symptoms. This study bridges prior theory and research concerning the psychological impact of normative developmental transitions, the developmental context of depression, and the associations among self-regulatory beliefs, achievement-related behavior, and emotional experience.

Achievement↗

Effect of egg storage upon the survival of Campylobacter jejuni in laboratory-infected fertile poultry eggs.

Fertile eggs were infected by Campylobacter jejuni in the laboratory by a temperature differential method of inoculation, which resulted in up to 10% of the hatched birds carrying C. jejuni in the intestine. When infected eggs were stored for 5 1/2 days before incubation, the infection rate of the eggs had decreased to 20% or less when set, and no infected chicks were hatched. Inoculation of eggs after 8 days in storage also failed to yield infected chicks. In all cases, the hatch ratio was no different from that of uninfected control eggs.

Animals↗