Presence of gamma glutamyl transferase in Mycobacterium smegmatis

The presence of gamma glutamyl transferase (GGT) has been established in Mycobacterium smegmatis. The 10,000 x g supernatant demonstrated only hydrolase activity and did not exhibit any transpeptidase activity. Most of the transferase activity was recovered in 100,000 x g supernatant demonstrating that GGT is a cytosolic enzyme. Maximum activity of GGT was observed at two days of growth and the activity decreased significantly till the seventh day of growth when mycobacteria was grown as stationary culture. The km for gamma glutamyl-p-nitroanilide was found to be 0.074 mM and Vmax for the reaction approached 11.9 nmol per min per mg protein. L-serine + borate was found to be a competitive inhibitor (Ki 12.05 mM) for GGT activity. The pH optimum for GGT activity was observed between 7.5 to 8.5 and temperature above 35 degrees C rapidly inactivated the enzyme activity. To the best of our knowledge, this is the first report which unequivocally establishes the presence of GGT activity in 10,000 x g supernatant of M. smegmatis.     

Reversibility of ultrastructural, contractile function and Ca2+ transport changes in guinea pig hearts after global ischemia

To understand the subcellular basis of contractile failure due to ischemia-reperfusion injury, effects of 20, 60, and 90 min of global ischemia followed by 30 min of reperfusion were examined in isolated guinea pig hearts. Cardiac ultrastructure and function as well as Ca2+ transport abilities of both mitochondrial and microsomal fractions were determined in control, ischemic, and reperfused hearts. Hearts were unable to generate any contractile force after 20 min of ischemia and showed a 75% recovery upon reperfusion. However, there were no significant changes in the subcellular Ca2+ transport in the 20-min ischemic or reperfused hearts. When hearts were made ischemic for 60 and 90 min, the recovery of contractile force on reperfusion was 50 and 7%, respectively. There was a progressive decrease in mitochondrial and microsomal Ca2+ binding and uptake activities after 60 and 90 min of ischemia; these changes were evident at various times of incubation period and at different concentrations of Ca2+. Mitochondrial Ca2+ transport changes were only partially reversible upon reperfusion after 60 and 90 min of ischemia, whereas the microsomal Ca2+ binding, uptake and Ca2+ ATPase activities deteriorated further upon reperfusion of the 90-min ischemic hearts. Ultrastructural changes increased with the duration of the ischemic insult and reperfusion injury was extensive in the 90-min ischemic hearts. These data show that the lack of recovery of contractile function upon reperfusion after a prolonged ischemic insult was accompanied by defects in sarcoplasmic reticulum Ca2+ transporting properties and structural damage.     

Effect of sub-diaphragmatic vagotomy on gastric mucosal mast cell population in pylorus ligated rats.

Gastric mucosal mast cell population was studied following sub-diaphragmatic vagotomy in albino rats, 6 and 12 h after pylorus ligation. Sub-diaphragmatic vagotomy significantly increased the gastric mucosal mast cell population in both 6 and 12 h groups, the increase being more in the latter. The results suggest that the vagal impulses act on the gastric mucosal mast cells causing their degranulation. Following vagotomy the contents stay bound within the mast cells. Increase in mast cell population with the longer experimental situation was possibly due to the continuation of normal turnover of the mast cells in the gastric mucosa. The present study, however, does not lead to a conclusion that the vagal influence on mast cell population is similar throughout the gastro-intestinal tract.     

An experimental model of ameboma in guinea pig.

Among the wide variety of clinicopathological manifestations of intestinal amebiasis, amebomas occur rarely and their pathogenesis is not well understood. When cholesterol-fed, 2- to 4-week-old guinea pigs were infected intracecally with a virulent, monoaxenic strain of Entamoeba histolytica, gross and histologically characteristic amebomas developed in 85% of the animals by the 3rd day, in 94% by the 9th day, and in 96% by the 12th day postinfection, by which time most of them had died. Amebomas were confirmed by histopathology. Thus, a model of consistent production of amebomas was documented.     

Studies on metabolism of vitamin A. The effect of the stage of vitamin A deficiency on sulphate activation in rat liver

1. The synthesis of ;active sulphate' in rat liver was studied at various stages of vitamin A deficiency, with the corresponding pair-fed controls. 2. The activity was significantly decreased even at the onset of the deficiency, and at the acute stage there was further loss. 3. Only at the earlier stages of the deficiency was the addition of retinol, in vitro, fully effective in restoring the lost activity; retinoic acid was partially active. No such restoration was possible at the acute deficiency stage.     

Effect of nifedipine administration on the functional capacity of neutrophils

Administration of nifedipine to mice over a period of six months caused a significant (p<0.05) decrease in neutrophilic functions viz superoxide generation, coupled to NADPH oxidase activity as well as NADPH production by HMP shunt. Properties like chemotaxis and phagocytosis showed a similar decrease. From this study, it is seen that nifedipine causes neutrophil functional abrogation which is therefore an apparent concern for the prolonged usage of the drug. However, relevance of the mouse model to clinical situation needs further investigation.