Altered Kinetics Properties of Erythrocyte Lactate Dehydrogenase in Type II Diabetic Patients and Its Implications for Lactic Acidosis.

Recent studies have been noted that the erythrocytes from Type II diabetic patients show significantly altered structural and functional characteristics along with the changed intracellular concentrations of glycolytic intermediates. More recent studies from our laboratory have shown that the activities of enzymes of glycolytic pathway changed significantly in RBCs from Type II diabetic patients. In particular the levels of lactate dehydrogenase (LDH) increased significantly.

Altered Erythrocyte Glycolytic Enzyme Activities in Type-II Diabetes.

The activity of enzymes of glycolysis has been studied in erythrocytes from type-II diabetic patients in comparison with control. RBC lysate was the source of enzymes. In the diabetics the hexokinase (HK) activity increased 50 % while activities of phosphoglucoisomerase (PGI), phosphofructokinase (PFK) and aldolase (ALD) decreased by 37, 75 and 64 % respectively but were still several folds higher than that of HK.

Hepatoprotective effect of withanolide-rich fraction in acetaminophen-intoxicated rat: decisive role of TNF-α, IL-1β, COX-II and iNOS.

Context: Overdose of acetaminophen (APAP) is common in humans and is often associated with hepatic damage. Withania somnifera (L.) Dunal (Solanaceae) shows multiple pharmacological activities including antioxidant and anti-inflammatory potential.

Evaluation of the bioavailability of major withanolides of Withania somnifera using an in vitro absorption model system.

Withania somnifera (L.) Dunal, shows several pharmacological properties which are attributed mainly to the withanolides present in the root. The efficacy of medicinally active withanolides constituents depends on the absorption and transportation through the intestinal epithelium.

Significance of antioxidant potential of plants and its relevance to therapeutic applications.

Oxidative stress has been identified as the root cause of the development and progression of several diseases. Supplementation of exogenous antioxidants or boosting endogenous antioxidant defenses of the body is a promising way of combating the undesirable effects of reactive oxygen species (ROS) induced oxidative damage. Plants have an innate ability to biosynthesize a wide range of non-enzymatic antioxidants capable of attenuating ROS- induced oxidative damage.

Effect of macronutrient deficiency on withanolides content in the roots of Withania somnifera and its correlationship with molybdenum content.

Context: The content of withanolides in the roots of Withania somnifera (L.) Dunal (Solanaceae) is important for therapeutic application. Earlier studies have shown that the deficiency of macro- and micronutrients affects the growth of W.

Effect of treatment with cadmium on structure-function relationships in rat liver mitochondria: studies on oxidative energy metabolism and lipid/phospholipids profiles.

Effects of treatment with a single intraperitoneal injection of cadmium (Cd) on oxidative energy metabolism and lipid/phospholipid profiles of rat liver mitochondria were examined at the end of 1 week and 1 month. Following Cd treatment the body weight increased only in the 1 month group, whereas the liver weight increased in both groups. State 3 and 4 respiration rates in general decreased significantly, with the maximum effect being seen with succinate.

Cadmium exposure-induced alterations in the lipid/phospholipids composition of rat brain microsomes and mitochondria.

Effects of treatment with a single intra-peritoneal (i.p.) injection (0.

Effect of treatment with cadmium on kinetic properties of Na(+), K(+)-ATPase and glucose-6-phosphatase activity in rat liver microsomes a correlative study on influence of lipid/phospholipid make-up.

Studies on Cd hepatotoxicity have focused mainly on induction of cytochrome P(450) system and related enzymes. In the present study young adult male rats given a single intra-peritoneal injection of Cd (0.84mg Cd/kg body weight) and effects on kinetic parameters rat liver microsomal Na(+), K(+)-ATPase and G6Pase were evaluated at the end of 1 month and 1 week.

Thyroidal regulation of substrate kinetics properties of cytochrome oxidase in rat liver mitochondria.

Effects of thyroidectomy (T(x)) and subsequent treatment with 3,5,3'-triiodothyronine (T(3)), and combined treatment (T(R)) with T(3) + thyroxine (T(4)) on substrate kinetics properties of cytochrome oxidase of rat liver mitochondria were examined. T(x) resulted in lowering of cytochromes content with decrease in the enzyme activity, and K(m) and V(max). T(3) and T(R) regimens restored the cytochromes contents and the V(max) values to normal.

Ageing-induced alterations in lipid/phospholipid profiles of rat brain and liver mitochondria: implications for mitochondrial energy-linked functions.

Effects of ageing on the lipid/phospholipid profile of brain and liver mitochondria from rats were examined. In the brain mitochondria the contents of total phospholipid (TPL) and cholesterol (CHL) increased with simultaneous increase in the TPL/CHL (mole:mole) ratio. The proportion and contents of lysophospholipid (Lyso), sphingomyelin (SPM), phosphatidylinositol (PI), phosphatidylserine (PS) and diphosphatidylglycerol (DPG) components increased, with maximal increases seen for PS and PI; phosphatidylcholine (PC) and phosphatidylethanolamine (PE) components registered decrease.

Thyroid hormone-induced alterations in membrane structure-function relationships: studies on kinetic properties of rat kidney microsomal Na(+),K (+)-ATPase and lipid/phospholipid profiles.

The effects of thyroidectomy (Tx) and subsequent treatment with 3,5,3'-triiodothyronine (T(3)) or combined replacement therapy (T(R)) with T(3 )and thyroxine (T(4)) on the substrate and temperature kinetics properties of Na+,K+-ATPase and lipid/phospholipid makeup of rat kidney microsomes were examined. Enzyme activity was somewhat high in the hypothyroid (Tx) animals and increased significantly following T(3) treatment, while T(R) treatment caused a decrease. In the Tx and T(3) groups enzyme activity resolved in two kinetic components, while in the T(R) group the enzyme showed allosteric behavior up to 0.

Diabetic modulation of the temperature kinetics properties of cytochrome oxidase activity in rat brain mitochondria.

The effects of alloxan-diabetes and subsequent treatment with insulin on temperature kinetics properties of cytochrome oxidase activity from rat brain mitochondria were examined. The enzyme activity decreased only at the late stage of diabetes which was not normalized by insulin treatment; however at early stage of diabetes hyper-stimulation occurred. In the control animals the Arrhenius plot was chair shaped with three energies of (E1, E2 and E3) and two phase transition temperatures (Tt1 and Tt2).

Dexamethasone treatment differentially affects the oxidative energy metabolism of rat brain mitochondria in developing and adult animals.

We studied the effect of repeated exposure to dexamethasone (Dex) treatment on rat brain mitochondrial oxidative energy metabolism in developing rats at different postnatal ages, i.e. 2-5 week and in adults.

Thyroid hormone treatments differentially affect the temperature kinetics properties of FoF1 ATPase and succinate oxidase as well as the lipid/phospholipid profiles of rat kidney mitochondria: a correlative study.

Effect of thyroidectomy (Tx) and subsequent treatment with 3,5,3'-triiodo-L: -thyronine (T(3)) or replacement therapy (T(R)) with T(3)+ L: -thyroxine (T(4)) on the temperature kinetics properties of FoF(1 )adenosine triphosphatase (ATPase, ATP synthase, H(+)-translocating ATP synthase EC 3.6.3.

Improved method for estimation of inorganic phosphate: implications for its application in enzyme assays.

The conventional method of Fiske and Subba Row for the estimation of inorganic phosphate (Pi) is although rapid, but suffers from the disadvantage that the color is unstable and hence the optical density (OD) measurements have to be carried out within a short time span of 8-12 min. This poses a restriction on the number of samples, which can be handled in a batch. Although, modified procedures involving use of alternate reducing agents/or increasing the concentration of H2SO4 in conventional method have been subsequently developed, but the problem of color stability could not be solved.

Effect of alloxan diabetes and subsequent insulin treatment on temperature kinetics properties of succinate oxidase activity in rat kidney mitochondria.

Early and late effects of alloxan diabetes and subsequent treatment with insulin on the temperature kinetics properties of succinate oxidase (SO) activity in rat kidney mitochondria were examined. In diabetic animals SO activity increased significantly and the increase was more pronounced at the late stage. Insulin treatment partially restored SO activity.

Diabetic cardiomyopathy and reactive oxygen species (ROS) related parameters in male and female rats: A comparative study.

Studies were carried out to examine and compare the effects of alloxan-diabetes on reactive oxygen species (ROS) related parameters in the heart from male and female rats. Effects of insulin treatment were also evaluated. The diabetic state severely compromised the ROS defense mechanism in the cardiac tissue and the effects were more pronounced in the female than in the male rats.

Stimulation of oxidative energy metabolism in liver mitochondria from old and young rats by treatment with dehydroepiandrosterone (DHEA). A comparative study.

Effects of treatment with DHEA (0.2 or 1.0 mg/kg body weight for 7 days) on oxidative energy metabolism of rat liver mitochondria from old (18-24 month old) and young (8-10 weeks old) male albino rats belonging to Charles-Foster strain were examined.

Insulin status-dependent alterations in lipid/phospholipid composition of rat kidney microsomes and mitochondria.

Early and late effects of alloxan-diabetes on lipid/phospholipid composition in rat kidney microsomes and mitochondria were examined. In microsomes, early diabetic state resulted in an increase in contents of total phospholipids (TPL), cholesterol (CHL), with an increase in the lysophospholipids (Lyso), phosphatidylcholine (PC), and phosphatidylinositol (PI) components. The sphingomyelin (SPM), phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidic acid (PA) content decreased.

Effect of alloxan-diabetes and subsequent treatment with insulin on kinetic properties of succinate oxidase activity from rat liver mitochondria.

We evaluated early and late effects of alloxan-diabetes and subsequent insulin treatment on the kinetic properties of succinate oxidase (SO) in rat liver mitochondria. Diabetic state lowered the SO activity; insulin treatment was effective in restoring the activity only in one-week diabetic rats. The energies of activation in low and high temperature ranges (EH and EL) decreased significantly in diabetic animals; once again insulin treatment was partially effective only in the one-week diabetic group.

Insulin-status-dependent modulation of FoF1 ATPase activity in rat kidney mitochondria.

The early and late effects of alloxan-diabetes and insulin treatment on kinetic properties of mitochondrial FoF1 ATPase were examined. Diabetic state resulted in significant decrease in the activity while insulin treatment caused hyper-stimulation. In control animals the enzyme activity resolved in three kinetic components.

Dehydroepiandrosterone treatment alters lipid/phospholipid profiles of rat brain and liver mitochondria.

Stimulation of mitochondrial function following treatment with dehydroepiandrosterone (DHEA) has been demonstrated. Since the activity of several electron transport chain components is dependent on specific lipid/phospholipid components, we examined the effects of DHEA treatment (0.1-2.

Insulin-status-dependent modulation of FoF1-ATPase activity in rat liver mitochondria.

Early and late effects of alloxan diabetes and insulin treatment on mitochondrial membrane structure and function were evaluated by studying the kinetic properties of mitochondrial membrane marker enzyme FoF1-ATPase and its modulation by membrane lipid/phospholipid composition and membrane fluidity. Under all experimental conditions the enzyme displayed three kinetically distinguishable components. In 1 wk-old diabetic animals the enzyme activity was unchanged; however, K(m) and V(max) of component I increased and K(m) of component II decreased.