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Biomedical subjects

M E Graichen

Publications and source records attributed to M E Graichen.

24 records · Page 2Linked to original sources
PubMed1981

Stability of activating systems for in vitro mutagenesis assays: enzyme activity and activating ability following long-term storage at - 85 degrees C.

Activating systems for in vitro mutagenesis assays are commonly prepared and stored at low temperature until required. The objective of the studies reported here was to determine the long-term stability of activating systems stored at - 85 degrees C. A broad range of microsomal enzymes in the postmitochondrial supernatant (PMS) and the microsomal fraction of livers from Aroclor 1254 treated rats were studied in conjunction with the ability of these fractions to catalyse the conversion of dimethylnitrosamine (DMN) and benzo(a)pyrene (B(a)P) to products mutagenic to Chinese hamster ovary (CHO) cells and Salmonella typhimurium TM677. Biphenyl-2- and biphenyl-4-hydroxylase showed a rapid decline in activity on storage, epoxide hydratase activity increased with storage and other enzyme activities studied were relatively stable for up to 32 weeks. No consistent trends in the ability of either the microsomes or the PMS to catalyze DMN or B(a)P induced mutation were observed for up to 12 weeks with CHO cells and 24 weeks with bacteria. It is concluded that low temperature storage of activating systems is an acceptable procedure. However, the results also indicate that certain enzyme activities change during storage, suggesting that aberrant results may be obtained when stored activating systems are used in in vitro tests to screen for mutagens.

Animals↗
PubMed1981

alpha-Naphthylisothiocyanate induced alterations in hepatic drug metabolizing enzymes and liver morphology: implications concerning anticarcinogenesis.

Alpha-naphthylisothiocyanate (ANIT) is a biliary toxin with anticarcinogenic properties. The studies described were designed to investigate the effects of continuous ANIT feeding on liver function. Male F-344 rats were fed ANIT at 0.01%, 0.022%, 0.047%, and 0.1% of the diet for 2, 4, and 6 weeks. Microscopic evaluation of liver sections revealed time- and dose- dependent bile duct proliferation, bile duct cell hypertrophy, and focal hepatocytic necrosis. Liver derived serum enzyme activity and serum bilirubin concentrations were increased in a fashion which correlated closely with the histological observations. A dose dependent decrease in hepatic cytochrome P-450 content, ethoxycoumarin-O-deethylase activity, and benzphetamine-N-demethylase activity was observed after 2 and 4 weeks of feeding ANIT. However, these enzyme activities returned to control values at 6 weeks in all except the 0.1% group. ANIT increased microsomal epoxide hydrolase and cytosolic DT-diaphorase activity (200-6005 of control). The enhancement was dose related and peaked at 2 and 4 weeks for epoxide hydrolase and DT-diaphorase, respectively. Both epoxide hydrolase and DT-diaphorase activity remained elevated at 6 weeks. These results suggest that ANIT mediated anticarcinogenesis, previously hypothesized to be the result of reduced mixed function oxidase activity, also may be accounted for by enhanced epoxide hydrolase and DT-diaphorase activity.

1-Naphthylisothiocyanate↗
PubMed1978

Kinetics and mechanism of dissociation of zinc ion from carbonic anhydrase.

The kinetics of dissociation of Zn2+ from the metalloenzyme carbonic anhydrase was measured over a range of pH, temperature, and acetate concentration. The rate of dissociation is extremely slow at neutral pH (t1/2 approximately 3) years, 4 degrees C), but increases in almost direct proportion to the hydrogen ion concentration and is enhanced in the presence of 1,10-phenanthroline or acetate. The thermodynamic stability of the zinc-apoenzyme complex was determined over a range of pH from rate data on binding and dissociation (stability constants 10(9)-10(11) M-1, 25 degrees C). The great stability of the complex and slow exchange of the apoenzyme ligand is attributed, at least in part, to the rigidity of the multidentate protein ligand.

Apoenzymes↗