Glutamate decarboxylase and GABA aminotransferase levels in different regions of rat brain on the onset of Leptazol induced convulsions.

The activities of Glutamate decarboxylase (GAD) and Gamma aminobutyric acid (GABA) were studied in three regions of rat brain in heightened neuronal activity resulting in convulsions by Leptazol. These enzymes were studied in preconvulsive, convulsive and post convulsive phases. The activity of GAD decreases significantly in the preconvulsive phase in all the three regions of brain followed by a significant increase during the convulsive and post convulsive phase in cerebral cortex and cerebellum. The activity of GABA-T decreases maximal during the preconvulsive phase followed by convulsive phase. The activity of this enzyme tended to increase to control values when the postconvulsive phase was reached. Therefore, it is suggested that the concomitant decrease of GAD activity and GABA concentration, is probably an important factor in the onset of convulsions.

Early oxidative change in low density lipoproteins during progressive chronic renal failure.

Modified low density lipoproteins (LDL), including their oxidized forms, have been widely implicated in the etiology of atherosclerosis and concomitant cardiovascular disease (CVD) in chronic renal failure (CRF). The nature of events that lead to oxidative changes in LDL proteins are not clearly understood. Thus, patients suffering from CRF were grouped into mild, moderate and severe categories based on their blood urea and serum creatinine levels. Progression of CRF was accompanied not only with gradual increase in serum malondialdehyde (MDA) but also parallel increase in conjugated diene and MDA levels in LDL fractions separated from serum. Serum superoxide dismutase (SOD) activity was concurrently found to decrease, along with a decrease in high-density lipoprotein (HDL) cholesterol, during the progression of CRF. Gradual increase in the appearance of LDL oxidation products seems to accompany progressive manifestation of CRF. The results presented suggest that determination of serum MDA and SOD levels may enhance the diagnostic significance of the study of lipid profile in determining the risk for cardio vascular disease in CRF.

Functional relationship between ammonia and gangliosides in brain.

The functional significance of ammonia production in brain under physiological or pathological conditions is not clearly known. NH4+ stimulates Na+, K+ activated ATPase causing stabilization of neuronal membranes of which gangliosides are major structural components. Moreover ammonia is known to inhibit lysosomal enzymes which include enzymes degrading gangliosides. Gangliosides have been shown to stimulate neuritogenesis in neuronal cultures and prevent the damage of the neurons from glutamate toxicity particularly in areas of brain ischemia. Hyperammonemia without any behavioural changes was induced in experimental rats by intraperitoneal administration of either a single dose (0.8 mmol/100 g wt.) or by six 'hourly' doses (0.6 mmol/100 g wt.) of ammonium acetate. An increase in the content of gangliosides along with a rise in the content of GD1A and GD1B without any change in beta-galactosidase and N-acetylhexosaminidase was observed in cerebral cortex, cerebellum, and brain stem, following the administration of single dose of ammonium acetate. Gangliosides, after extraction from the different brain regions, were estimated by the thiobarbituric acid method and expressed in terms of sialic acid. Individual gangliosides were separated and estimated by thin layer chromatography using resorcinol as the staining agent. These results suggest that ammonia production in the neuronal pathways in brain either as a result of repeated stimulation under physiological conditions or as a result of focal ischemia or injury, may likewise cause an increase in the content of gangliosides which may help in neuritic growth (physiological conditions facilitating synaptic plasticity) and may exert a protective effect on the neurons in the ischemic area against glutamate toxicity.

Acute and long-term effects of chlorpromazine on glutamine synthetase and glutaminase in rat brain.

The effect of administration of chlorpromazine on the activity of glutamine synthetase and glutaminase and the content of glutamate and gamma-aminobutyric acid (GABA) in different regions of rat brain was studied in an investigation of the possible role of these amino acids in the lowering of the seizure threshold following prolonged administration of chlorpromazine. Chlorpromazine was administered at a dose of 20 mg/kg of body weight s.c. For the acute study, the animals were killed 20 min after a single injection. For the long-term study, the animals were treated every day with the same dose for 21 days and were killed 20 min after the last injection. The results showed an increase in glutamate level in each brain region investigated following long-term administration, but only in the cerebral cortex after a single dose. GABA levels showed an increase in the brainstem only in acute experiments. Glutamine synthetase activity was increased in all three regions after a single dose and only in cerebral cortex after long-term administration. Glutaminase activity showed a decrease in cerebral cortex only after long-term administration of the drug. These results suggest the possible occurrence of a state of increased excitability in the brain as a result of long-term administration of chlorpromazine, thus contributing to the known complication of seizures.

Serum malanodialdehyde (TBA reactive substance) levels in cigarette smokers.

Serum malanodialdehyde (MDA) levels in 40 smokers and 23 non-smokers belonging to different age groups were estimated. The MDA levels were high in smokers of all age groups having a history of smoking for less than 10 years. MDA levels, however, were not elevated in smokers with a history of smoking for more than 10 years. These results are discussed and are interpreted as suggestive that MDA might alter the LDL and lead to deposition of cholesterol in arterial wall macrophages explaining thereby the risk of ischaemic heart disease in cigarette smokers.

Acute and short-term effects of lithium on glutamate metabolism in rat brain.

Amino acids of the glutamate family, viz. glutamic acid, aspartic acid, glutamine, gamma-amino-butyric acid (GABA) and alanine, along with the activities of glutamic acid dehydrogenase (GDH), aspartic acid aminotransferase (AST), alanine aminotransferase (ALT), glutamine synthetase (GS), glutaminase, glutamic acid decarboxylase (GAD) and GABA-aminotransferase (GABA-T) were estimated in cerebral cortex, cerebellum and brain stem of rats treated with a single dose of lithium or with seven daily doses of lithium (3 m-equiv./kg body wt). The levels of GABA were found to increase in cerebral cortex and brain stem following the administration of a single dose and also were found to be increased in cerebral cortex and cerebellum after treatment for 7 days. The content of glutamic acid was increased in all three brain regions after treatment for 7 days. Glutamine was increased in both cerebral cortex and brain stem after treatment for 7 days, whereas aspartic acid was increased in brain stem after both the administration of single dose and treatment for 7 days. A significant increase (P less than 0.05) in the activity of GS was observed in brain stem after 7 days of treatment. Similarly, a significant increase (P less than 0.01) in the activity of AST was observed in all three regions of the brain following the treatment for 7 days. The above results are discussed in relation to the known effects of lithium on brain cation metabolism and a suggestion is made that an imbalance in the functional activities of glutamic acid and GABA as a result of quantitative changes in these amino acids, brought about by lithium, may play a role in the therapeutic efficacy of lithium in bipolar disorders.

Acute metabolic effects of taurine on the enzymes metabolizing glutamate and gaba.

100 mg of taurine per kg body weight had been administered intraperitoneally and 30 min after the administration the animals were sacrificed. Glutamate dehydrogenase, aspartate aminotransferase, alanine aminotransferase, glutaminase, glutamine synthetase, glutamate decarboxylase and GABA aminotransferase along with the content of glutamate and GABA in cerebral cortex, cerebellum and brain stem were studied and compared with the same obtained in the rats treated with normal saline in place of taurine. The results indicated a significant decrease in the activity of glutamate dehydrogenase in cerebral cortex and cerebellum and a significant increase in brain stem. Glutaminase and glutamine synthetase were found to increase significantly both in cerebral cortex and cerebellum. The activities of glutamate decarboxylase was found to increase in all the three regions along with a significant decrease in GABA aminotransferase while the content of glutamate showed a decrease in all the three brain regions, the content of GABA was observed to increase significantly. The above effects of taurine on the metabolism of glutamate and GABA are discussed in relation to the functional role of GABA and glutamate. The results indicate that taurine administration would result in a state of inhibition in brain.

Acute and short term effects of ethanol on membrane enzymes in rat brain.

Changes in the biophysical and biochemical character of membranes brought about by ethanol have been emphasized in the underlying mechanism of alcohol toxicity. Membrane enzymes such as Na+, K+ activated ATPase, 5'-nucleotidase, and gamma-glutamyl transpeptidase were studied in cerebral cortex, cerebellum, and brain stem of rats subjected to acute and short term ethanol toxicity. Acute ethanol toxicity was induced by intraperitoneal injection of 1 ml of 7M ethanol per 100 g body weight of rat and the animals were sacrificed half an hour after the administration. Short term ethanol toxicity was induced by intraperitoneal injections of 0.5 ml (7 M ethanol) per 100 g weight of the rat for 7 days and the animals were sacrificed half an hour after the last injection. In acute ethanol toxicity the activity of Na+, K+-activated ATPase was found to decrease significantly in cerebral cortex and brain stem, while in short term alcohol toxicity, the activity was found to increase in cerebral cortex and cerebellum. The activity of gamma-glutamyl transpeptidase was found to increase in all the three regions in acute and short term ethanol toxicity. No change in the activity of 5'-nucleotidase was observed in any of the regions either in acute or in chronic ethanol toxicity. While a significant increase in the activity of adenosine deaminase was found in cerebral cortex, cerebellum, and brain stem in acute ethanol toxicity, the same was found to decrease significantly in cerebral cortex and a persistent increase in brain stem in short term ethanol toxicity. The above changes in the activities of the enzyme were discussed with reference to the well known changes in the membrane structure and consequent alteration in brain function.

Functional relationship of ammonia to DNA, RNA and protein in brain.

The functional significance of the production of ammonia during neuronal activity is not known, although a number of studies are available concerning the mechanism of of ammonia toxicity in brain. Hyperammonemia without behavioural change was induced in experimental rats by i.p. administration of either a single dose of ammonium acetate or 6 doses with an interval of 1 h during the day or one single dose daily for 7 days. No significant change was observed in the content of DNA, RNA and protein in cerebral cortex, cerebellum and brain stem following the administration of a single dose. There was a marked decreased in the content of DNA in all three brain regions after the administration of 6 hourly doses of ammonium acetate. Under these conditions the RNA was found to increase in all three brain regions. A decrease in the content of protein per gram wet weight was observed in cerebral cortex and brain stem while the change in cerebellum was found to be insignificant. However, the content of RNA and protein was found to show a very significant increase when expressed per microgram of DNA for each region under these conditions, indicating an increase in RNA and protein in the cells surviving the toxic damage caused by ammonia. A marked and significant increase in the content of RNA and protein was observed in all three brain regions following the administration of a single dose per day for 7 days. These results are discussed not only with respect to toxic damage to the glial cells by ammonia, but also with respect to the probable effect of ammonia liberated during neuronal activity on inhibition of lysosomal degradation of protein, and on its stimulatory effects of transcriptional and translational activities. The increased RNA and protein under these conditions may be contributing not only in the proliferation of glial cells (Alzheimer Type II cells) but also in the dendritic spinous growth.

Functional relationship of ammonia with RNA, DNA, and protein in rat muscle.

The acid proteinases, neutral proteinases, protein, RNA, and DNA were estimated along with the ammonia content in muscle in rats subjected to experimental hyperammonemic states (acute and chronic) by the administration of ammonium acetate intraperitoneally. It was observed that there was a decrease in the activities of acid proteinases and neutral proteinases in muscle in acute as well as in chronic hyperammonemic states. A rise in protein content was observed in muscle under these conditions. No changes were seen in the DNA content while RNA showed a slight increase in muscle. These results were discussed with reference to the possible role of ammonia in lysosomal protein degradation as well as to its probable effect on protein synthesis through its action on transcription.

Acute and short term effects of ethanol on the metabolism of glutamic acid and GABA in rat brain.

Although alcoholic intoxication is attributed to its pharmacological effects on the cell membranes in brain, the rapid metabolic utilisation of the same alters the metabolism of brain affecting the metabolism of glutamate and GABA which have varied metabolic roles besides serving a major proportion of synaptic activity. A study on the effects of ethanol, both acute and short-term, on glutamate (glu) and GABA metabolism in various regions of rat brain was carried out. Increased activities of glutamic acid decarboxylase (GAD) and aspartic acid aminotransferase (AST) in all brain regions, but decreased activity of glutamic acid dehydrogenase (GDH) in cerebral cortex (CC) and cerebellum (CB) following ethanol administration in brain was observed. Differential effects of ethanol were also obtained on the contents of glu and aspartate (asp), which were increased in CC, CB, and brain stem (BS) regions, as opposed to GABA content, which, although found to increase in acute toxicity, showed a decrease in all of the above brain regions in short-term toxicity. It is concluded that the above changes in glu, asp and GABA represent the consequences of metabolic utilization of alcohol in the brain, probably more a state of cerebral excitation than depression, and the changes may be a compensatory phenomenon.

Acute effects of ethanol on production & disposal of adenosine from rat myocardium.

Adenosine is a local hormone and is considered to act as a vasodilatory substance when released locally. Alcohol is known to affect membrane structure and acts as a coronary vasodilator. Membrane enzymes such as 5'-nucleotidase, adenosine deaminase, and gammaglutamyl transpeptidase, along with AMP deaminase, have been studied in rat myocardial tissue following the administration of a sufficiently toxic dose (producing semiconsciousness) of ethanol (1ml of 7M ethanol/100g body wt.). The activity of 5'-nucleotidase as well as that of adenosine deaminase increased due to the administration of ethanol, without any significant change in the activities of gammaglutamyl transpeptidase and AMP deaminase. These changes are discussed in relation to the metabolic changes occurring in the myocardium and the resultant effects on the coronary vessels.

Aromatic L-amino acid decarboxylase, dopamine beta-hydroxylase, monoamine oxidase, malondialdehyde, and acid phosphatase in rat brain capillaries and kidney glomeruli in experimental hypertension.

Some metabolic consequences of experimental hypertension on rat brain capillaries and kidney glomeruli have been studied in rats made hypertensive by a combination of deoxycorticosterone acetate injection and elevated salt intake (DOCA-salt hypertension) and isoproterenol injection. Enzyme activities were studied in vitro to ascertain directly or indirectly any changes in the metabolism of catecholamines and prostaglandins, and lysosomal integrity under conditions of experimental hypertension. Experimental hypertension was accompanied by an elevation in the activities of aromatic L-aminoacid decarboxylase, dopamine beta-hydroxylase, and malondialdehyde concentration, both in the brain capillaries and kidney glomeruli of rats. On the other hand, monoamine oxidase activity increased in brain capillaries but decreased in kidney glomeruli. Acid phosphatase activity increased marginally in kidney glomeruli but decreased significantly in brain capillaries. The catecholamine-synthesizing potential appears to have been augmented in both the tissue capillaries with a compensatory increase in the degrading enzyme activity in the brain capillaries of hypertensive rats. The absence of such an increase and an actual decrease in the monoamine oxidase activity in the kidney glomeruli may be responsible for the sustained maintenance of the hypertensive state. Increased malondialdehyde concentration may be due to the stimulation of the prostaglandin metabolism by the augmented catecholamine metabolism.

Activities of glutamine synthetase, glutaminase and arginase in kainic acid lesioned rat brain corpus striatum.

Intrastriatal kainic acid (2 ?g/?l) administration gave rise to significant increase in activities of glutamine synthetase and arginase along with a significant decrease in the activity of glutaminase in the lesioned striatal tissue 7 days after the administration of kainic acid. The increase in the activity of glutamine synthetase was attributed to the gliosis occurring in such lesions. The decrease in the activity of glutaminase was thought to be due to the loss of GABAergic neurons. The increase in arginase activity might be occurring in glial cells or in nerve endings. Although the earlier results indicated a low specific activity of arginase in glial cells, the observed increase in its activity might be partly due to its increase in proliferating glial cells, liberating ornithine for the formation of polyamines. However, it was also thought that a substantial increase may be occurring in the arginase present in the intact glutamatergic (corticostriate pathway) nerve endings, since it was earlier found that the synaptosomes of the rat brain had appreciably high activity of arginase. These results were discussed in relation to the probable roles of arginine and glutamine as the precursors for neurotransmitter pools of glutamate in striatum.

Functional significance of the activities of glutaminase and ornithine-?-aminotransferase in rat brain.

The activities of glutaminase, glutamine synthetase (GS), arginase and ornithine amino transferase (orn-T) were studied in three regions of rat brain in heightened neuronal activity by producing convulsions by leptazol. These enzymes were studied in preconvulsive, convulsive and postconvulsive phases. Glutaminase activity was found to increase in all the three regions in the preconvulsive and convulsive phases. GS activity decreased in the preconvulsive phase but rose gradually to the control level when the postconvulsive phase was reached. The activity of arginase decreased in the cerebellum in preconvulsive and convulsive phases. However, in the cerebral cortex there was a decrease in the activity of this enzyme only in the convulsive phase. The results suggest that glutamine acts more likely as a precursor for the neurotransmitter pool of glutamate, while ornithine serves more as a precursor for the neurotransmitter pool of GABA.

Possible occurrence of ornithine-omega-aminotransferase in GABAergic neurons.

The specific precursors for neurotransmitter pools of glutamate giving rise to GABA in GABAergic neurons and nerve endings have not been clearly established. Glutamate is the immediate precursor for the production of GABA and it is suggested that ornithine (from arginine) might be serving as one of the precursors of glutamate for the formation of neurotransmitter pool of GABA. Damage to GABAergic neurons in different regions of the brain in anoxia is well known. If arginine and ornithine act as precursors for GABA in GABAergic neurons, a decrease in the activities of arginase and ornithine-omega-transferase (Orn-T) is possible in areas having the lesions involving the GABAergic neurons due to anoxia. Estimation of Orn-T and arginase in different regions of the brain of rats exposed to anoxia revealed such a possibility.

Adenosine deaminase in convulsions along with its regional, cellular and synaptosomal distribution in rat brain.

Adenosine deaminase activity was determined in enriched neuronal, glial and synaptosomal fractions in three regions of rat brain. It was found to be equally distributed in cortical neurons and synaptosomes. Appreciable activity was observed in glial cells. A significant increase in the activity of this enzyme was observed in three brain regions in rats subjected to leptazol convulsions. No significant change, however, was found in the pre- or post-convulsive periods. These results are discussed in relation to the possible degradation of adenosine, a neuroinhibitory substance and to the possible production of inosine, an initiator of seizure activity in low amounts and depressor of the same in high amounts.

Intracellular content of thiol compounds, thiobarbituric acid reactive substances and gamma-glutamyl transpeptidase in rat brain during anoxia.

Intracellular thiol compounds like glutathione play an important role in scavenging free radicals. Free radical reactions occur in the respiratory chain, and free radicals are known to accumulate in hypoxia and ischaemia. The content of thiobarbituric acid (TBA) reactive substances, thiol and non-protein thiol (reduced glutathione) concentrations along with the activity of gamma-glutamyl transpeptidase (GGTP), a membrane enzyme, were estimated in three regions of the brain in rats exposed to anoxia. It was observed that no change occurred in the content of TBA-reactive substances in any one of the three brain regions, although a significant increase in the non-protein thiol concentration occurred uniformly during anoxia. A significant increase in GGTP occurred in all the three brain regions during anoxia. The increase in the activity of GGTP is suggestive of an alteration in the function of blood-brain barrier during anoxia. These results are suggestive of an absence of lipid peroxidative damage during anoxia.

Effect of simultaneous administration of vitamin E and pyridoxine on erythrocyte membrane Na+K+ ATPase activity.

Vitamin E is known to have an interaction with pyridoxine. To understand the functional significance of this interaction a study was carried out on the effect of simultaneous administration of vitamin E and pyridoxine on erythrocyte membrane Na+K+ ATPase activity. Adult male volunteers belonging to low socio-economic class with prior consent were used as subjects for the study. The results of the study have shown that administration of vitamin E alone brings about a transient increase in the total and ouabain insensitive Na+K+ ATPase. Administration of B6 along with vitamin E brought about an increase in the total and true Na+K+ ATPase activity. B6 administration alone showed a statistically insignificant increase in true ATPase activity. The results of the study are interpreted to show that administration of vitamin E along with B6 is beneficial to membrane function.