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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. The 1 week Cd group showed decreased respiratory activity with glutamate, pyruvate + malate, and succinate as the substrates. In the 1 month Cd-treated group respiration rates recovered with glutamate and pyruvate + malate but not with succinate. All cytochrome contents decreased in the 1 week Cd-treated group but recovered in the 1 month group. ATPase activity registered an increase in both Cd-treated groups. Dehydrogenase activities increased in the 1 week group but decreased in the 1 month Cd-treated group. The mitochondrial cholesterol content increased in the 1 week Cd-treated group. In the 1 week Cd-treated group the lysophospholipid (Lyso), sphingomyelin (SPM), and diphosphatidylglycerol (DPG) components increased. By contrast, the phosphatidylethanolamine (PE) component decreased. In the 1 month Cd-treated group the phosphatidylinositol, phosphatidylserine, and DPG components increased, whereas the Lyso, SPM, and phosphatidylcholine components decreased. The results demonstrate that single-dose Cd treatment can have adverse effects on liver mitochondrial oxidative energy metabolism and lipid/phosphopholipid profiles, which in turn can affect membrane structure-function relationships. [ABSTRACT FROM AUTHOR]

Abstract: Effects of treatment with a single intra-peritoneal (i.p.) injection (0.84mgCd/kg body weight) of cadmium (Cd) on lipid/phospholipids profiles of rat brain microsomes and mitochondria were examined. At the end of one-week following treatment with Cd the microsomal total phospholipids (TPL) content was unchanged but the cholesterol (CHL) content increased. In one-month Cd-treated group both TPL and CHL contents increased. In one-week Cd-treated group the content of phosphatidylinositol (PI) decreased whereas that of lysophospholipid and phosphatidic acid (PA) increased. In one-month Cd-treated group the sphinghomyelin (SPM) and phosphatidycholine (PC) components increased whereas phosphatidyethanolamine (PE) and PA decreased. In mitochondria, CHL content increased in both Cd-treated groups. The content of TPL decreased only in one-month Cd-treated group. In one-week Cd-treated group the lysophospholipids increased whereas PI and phosphatidylserine (PS) decreased. In one-month group the lysophospholipids, PI and diphosphatidylglycerol (DPG) components decreased whereas PS and PE component increased. The results suggest that exposure to a single dose of Cd has differential and long-lasting effects on lipid/phospholipids profiles of rat brain microsomes and mitochondria. [Copyright &y& Elsevier]

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. In the liver mitochondria contents of TPL and CHL increased. However, the TPL/CHL (mole:mole) ratio was not altered. Lyso, PI and PS increased. However, the magnitude of increase was competitively lower; PE and DPG decreased. SPM and PC did not change as a consequence of ageing. These changes altered the contents of individual phospholipids in the two membrane systems. Respiration with glutamate, pyruvate + malate, succinate and ascorbate + N,N,N',N'-tetramethyl-p-phenylenediamine was significantly impaired in brain mitochondria from old animals. For liver mitochondria the respiratory activity declined with glutamate and succinate. Correlation studies by regression analysis revealed that the lipid/phospholipid classes regulate respiratory function differently in the mitochondria from the two tissues. The respiration-related parameters in the brain mitochondria were dependent on multiple lipid/phospholipid components, and the process of regulation was complex compared to the liver mitochondrial functions. [ABSTRACT FROM AUTHOR]

The effects of thyroidectomy (Tx) and subsequent treatment with 3,5,3′-triiodothyronine (T3) or combined replacement therapy (TR) with T3 and thyroxine (T4) 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 T3 treatment, while TR treatment caused a decrease. In the Tx and T3 groups enzyme activity resolved in two kinetic components, while in the TR group the enzyme showed allosteric behavior up to 0.5 m m ATP concentration. The K m and V max values of both the components decreased in Tx animals without affecting the catalytic efficiency. T3 treatment caused a significant increase in the V max of both the components, with a significant increase in the catalytic efficiency, while the K m values were not upregulated. The TR regimen lowered the K m and V max of component II but improved the catalytic efficiency. Thyroid status-dependent changes were also noted in the temperature kinetics of the enzyme. Regression analysis revealed that changes in the substrate and temperature kinetics parameters correlated with specific phospholipid components. [ABSTRACT FROM AUTHOR]

Abstract: Objectives: : Effects of treatment with dehydroepiandrosterone (DHEA) on oxidative energy metabolism in rat liver and brain mitochondria were examined. Design and methods: : Young adult rats were administered DHEA (0.1, 0.2, 1.0 or 2.0 mg/kg body weight) by subcutaneous route for 7 consecutive days. Results: : DHEA treatment resulted in general, in stimulation of state 3 respiration rates without having any uncoupling effect on ADP/O ratios. The stimulation of state 3 respiration rate for a given substrate was dose dependent in a tissue-specific manner. Parallel increases in the contents of cytochromes aa3 and b were also noted. DHEA treatment stimulated the glutamate dehydrogenase (GDH) and succinate DCIP reductase (SDR) activities. Under the treatment conditions, mitochondrial ATPase activity was also stimulated. Conclusions: : Treatment with DHEA significantly stimulated oxidative energy metabolism in liver and brain mitochondria. [Copyright &y& Elsevier]

Abstract: Effects of treatment with dehydroepiandrosterone (DHEA) (0.2 or 1.0mg/kg body weight for 7 days) on oxidative energy metabolism in cerebral mitochondria from developing and young adult rats were examined. Treatment with DHEA did not change the body weight of developing rats but resulted in increase in the brain weight in 5 week group. In young adult rats the body weight increased following treatment with 1.0mg DHEA. State 3 and state 4 respiration rates with all the substrates increased following DHEA treatment, the effect being more pronounced in the developing rats. State 4 respiration rates were stimulated to variable extents. Contents of cytochromes aa 3 and b increased following DHEA treatment and once again the effect was more pronounced in the developing rats. DHEA treatment marginally changed the content of cytochromes c + c 1. In the developing rats the ATPase activity and the levels of dehydrogenases increased significantly by DHEA treatment. Results of our studies have shown that treatment with exogenous DHEA accelerates the process of maturation of cerebral mitochondria thus emphasizing the role of DHEA in brain development in postnatal life. [Copyright &y& Elsevier]

Abstract: The content of the neurosteroids, dehydroepiandrosterone (DHEA) in the brain decreases with aging. Also the oxidative energy metabolism is known to decrease with aging. Hence we examined the effects of treatment with DHEA (0.2 or 1.0mg/kg body weight for 7 days) on oxidative energy metabolism in brain mitochondria from old and young adult rats. State 3 respiration rates in brain mitochondria from old animals were considerably lower than those in young adults. Treatment with DHEA stimulated state 3 and state 4 respiration rates in both the groups of the animals in a dose-dependent manner. In the old rats following DHEA treatment, the state 3 respiration rates became comparable to or increased beyond those of untreated young adults. In contrast to the old rats, stimulatory effect of DHEA treatment was of greater magnitude in the young adults. However, at higher dose (1.0mg) the effect declined. Cytochrome aa 3 content in the brain mitochondria from old rats was significantly low but the content of cytochrome b was unchanged while the content of cytochromes c + c 1 had increased. Treatment with DHEA increased the content of cytochrome aa 3 and b in old as well as in young adult animals. Higher dose of DHEA (1.0mg) had adverse effect on the content of cytochrome c + c 1 . DHEA treatment stimulated ATPase activity in a dose-dependent manner in young adult rats whereas in the old rats the effect on ATPase activity was marginal. Dehydrogenases activities were somewhat lower in the old rats. DHEA treatment stimulated mitochondrial dehydrogenases activities in both the groups. Results of our studies suggest that judicious use of DHEA treatment can improve oxidative energy metabolism parameters in brain mitochondria from young adult as well as old rats. [Copyright &y& Elsevier]

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. In diabetic condition only component I and II were present. With insulin treatment component III was restored but component II was abolished. Diabetic state and insulin treatment had varied effects on Km values of the three components, whereas the Vmax values were generally on the higher side. Evaluation of the AppKcat/Km values revealed that diabetic state resulted in increased catalytic efficiency; insulin treatment brought back these values to normality. Temperature kinetics studies indicated that the phase transition temperature decreased significantly in the diabetic and insulin-treated diabetic animals. The energy of activation in low temperature range increased in the diabetic animals. Insulin treatment corrected the Arrhenius pattern at early stage of diabetes; at late stage the pattern was reversed. The results are suggestive of subtle insulin-status-dependent alterations in membrane structure – function relationships. [ABSTRACT FROM AUTHOR]

Abstract: Early and late effects of alloxan-diabetes of lipid/phospholipid composition of rat brain microsomes and mitochondria were examined. In microsomes, early as well as late diabetic stages resulted in decrease in contents of total phospholipids (TPL) and increase in cholesterol (CHL). Insulin treatment restored TPL with further increase in CHL in 1 week group. In early diabetic stage there was increase in the sphingomyelin (SPM) while phosphatidylinositol (PI) and phosphatidylserine (PS) components decreased. Insulin treatment restored SPM and decreased the lysophospholipids (Lyso), PI, PS and phosphatidic acid (PA); phosphatidylethanolamine (PE) increased. In 1 month diabetic group phosphatidylcholine (PC) decreased while PI, PS and PE increased. Insulin treatment lowered the Lyso, SPM, PI, PS and PA while PC and PE increased. In mitochondria, at early stage of diabetes both CHL and TPL contents decreased; insulin treatment restored the former component. Late diabetic stage had no effect on CHL and TPL contents; insulin treatment brought about reduction in both. Diabetic state had marginal effect on phospholipid composition at both the stages. Insulin treatment had a generalized effect of lowering of PI and PS components and increasing diphosphatidylglycerol (DPG). The fluidity of microsomal membranes decreased progressively in the diabetic condition; insulin treatment fluidized the membrane at early stage. The fluidity of mitochondrial membranes increased in early diabetic stage and the effect was accentuated by insulin treatment. However, at the late stage the effects on membrane fluidity were marginal. [Copyright &y& Elsevier]

Abstract: Effects of STZ diabetes and treatment with insulin on cerebral mitochondrial metabolism in the male and female rats were examined. Diabetic state resulted in generalized decrease in the state 3 respiration rates in the males with practically all the substrates except glutamate where the opposite effect was seen. Diabetic state had no adverse effect on the respiratory activity in the females. Insulin treatment had no restorative effect in the males. By contrast in the females, adverse effects were noted. The cytochromes contents decreased in STZ diabetes with the effect being more pronounced in the males; treatment with 1 unit of insulin restored the cytochromes contents. STZ diabetes also resulted in decreased dehydrogenases activities with the effect being more pronounced in the females: insulin treatment resulted in hyper-stimulation of glutamate dehydrogenase and succinate DCIP reductase activities; restoration of malate dehydrogenase activity was only partial. The results point out that STZ diabetes and insulin treatments differentially affect cerebral mitochondrial energy metabolism in the male and female rats. [Copyright &y& Elsevier]

Abstract: Effect of neonatal hypothyroidism on kinetic properties of Ca2+-ATPase from rat brain synaptic plasma membranes and microsomes were examined. Neonatal hypothyroidism resulted in significant decrease in the enzyme activity in both the membrane systems. The synaptic membranes in control group displayed presence of one kinetic component whereas a low affinity component became evident in the hypothyroid group. In the microsomes, both control as well as hypothyroid groups showed presence of two kinetic components with the latter group showing two-fold increase in the K m. The Ca2+ binding characteristics were generally unaltered in the enzyme from both the membrane systems. Our results suggest that impairment in the Ca2+-ATPase activity together with altered kinetic properties could be one of the underlying biochemical mechanisms leading to CNS dysfunction as a consequence of thyroid hormone deprivation during critical stages of brain development. [Copyright &y& Elsevier]

Abstract: Cerebral microsomal membrane properties were assessed in the chronic condition of generalized seizure induced by picrotoxin (PTX) in rats. PTX-induced seizures resulted in increased lysophosphatidyl glycerol, phosphatidylcholine and phosphatidic acid components, while phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol were significantly reduced by 19–73%. The cholesterol (CHL) content increased considerably by 25% without alteration in total phospholipids content. Microsomal membrane was more fluidized in the epileptic condition. Possible consequences of microsomal membrane alterations are discussed in terms of deregulation of Ca2+ homeostasis. In conclusion, alterations in the microsomal membrane properties may have a significant influence on the cerebral function in the chronic epileptic condition. [Copyright &y& Elsevier]

Aim: The incidence of coronary heart diseases (CHD), congestive heart failure (CHF) and myocardial infarction is higher in diabetic patients than in non-diabetic groups, with these incidences being more in women than in the men. Hence, we examined involvement of mitochondrial energy transduction functions. Methods: Mitochondrial energy metabolism in cardiomyopathy was studied using streptozotocin (STZ)-diabetic male and female rats as the model system. Effects of insulin treatment were also evaluated. Results: The body and heart weights decreased in both male and female diabetic rats. Insulin treatments resulted in significant increase in the body and heart weights in the female rats. Mitochondrial respiration rates with all the substrates tested decreased in diabetic condition in both males and females. Treatment with two dose -regimens of insulin had differential restorative effect on mitochondrial substrate oxidation in the males but caused hyper-stimulation in the females. Diabetic state brought about 19% decrease in the cytochrome aa3 content in the female rats. Treatment with 0.6 units of insulin significantly increased the cytochrome contents in general in both the sexes whereas higher dose (1.0 unit) caused decrease in the cytochromes content in the females. Diabetic state resulted in decreased dehydrogenases activities; insulin treatments had differential effect on the dehydrogenase activity in the males and the females. The results suggest that insulin treatment-induced hyper-stimulation of respiration in female rats may lead to increased production of reactive oxygen species. Besides, increased formation of advanced glycosylated end products may further lead to increased risk of CHF and CHD. Conclusions: The results suggest that differential effects of STZ-diabetes and insulin treatments in the female rats than in males may be the underlying cause for increased incidence of diabetic cardiomypathies in the females. [ABSTRACT FROM AUTHOR]

The current study examined the ability of antimalarials chloroquine (CQ), primaquine (PQ), and quinine (Q) to inhibit human erythrocyte membrane acetylcholinesterase (AChE) and the mechanisms underlying their inhibitory action. CQ was found to be the most effective inhibitor of the enzyme followed by PQ and Q. The concentrations required to obtain 33% inhibition (IC 33 ) for CQ and PQ were 22 and 38 µM, respectively, whereas that for Q was 3.2 mM. The concentrations required to obtain 67% inhibition (IC 67 ) were about 9 and 7 times higher for CQ and PQ, whereas that for Q was only about 2.5 times higher. Hill plot analysis revealed that CQ shows de-binding above 40 µM. The two kinetic components of AChE were inhibited by the three antimalarials, and the inhibition was of mixed type. Increasing concentrations of antimalarials caused progressive decrease in the V max of both components. IC 33 concentrations resulted in 1.6- to 6-fold increase in K m of both the components while IC 67 concentration caused 2.8- to 13-fold increases in K m with maximum effect being seen with Q. The K i values were lowest for CQ suggesting that it was the most potent inhibitor; these values were 3.3 and 60 times higher for PQ and Q. Antimalarials represent the bifunctional compounds that possess anti-inflammatory properties and also inhibit cholinesterases. The results of our studies suggest that 4-aminoquinoline–based antimalarials like CQ and hydroxychloroquine, which are both potent anti-inflammatory agents and inhibitors of cholinesterases, may have potential use as the most effective neuroprotective agents against amyloid-β-peptide (Aβ) neurotoxicity in Alzheimers disease. [ABSTRACT FROM AUTHOR]

Abstract: Effects of treatment with thyroid hormones l-3,5,3′-tri-iodothyronine (T3) and l-thyroxine (T4) on oxidative energy metabolism in cerebral mitochondria from hypothyroid adult rats were examined. It was observed that T3 and T4 stimulated respiratory activity in a substrate-specific and dose-dependent manner. The results also suggest that the synthesis of cytochrome aa 3 and of cytochrome c may be dependent on both T3 as well as T4 whereas higher concentrations than normal of T3 and T4 may have a catabolic influence on cytochrome b content. [Copyright &y& Elsevier]

Abstract: Mitochondrial function is a key determinant of both excitability and viability of neurons. Present studies were carried out to decipher cerebral mitochondrial oxidative energy metabolism and membrane function in the chronic condition of generalized seizures induced by picrotoxin (PTX) in rats. PTX-induced convulsions resulted in decreased respiration rates (14–41%) with glutamate, pyruvate + malate, and succinate as substrate. The ADP phosphorylation rates were drastically reduced by 44–65%. An opposite trend was observed with ascorbate + N,N′,N′-tetramethyl-p-phenylenediamine (TMPD) as substrate. In general, uncoupling of the mitochondrial electron transport was observed after PTX treatment. Malate dehydrogenase (MDH) and succinate dehydrogenase (SDH) activities were decreased by 20–80%; also, there was significant reduction in cytochrome b content after PTX treatment, while the FoF1 ATPase (complex V) activity increased in basal and 2,4-dinitrophenol (DNP)-stimulated condition, indicating increased membrane fragility. The substrate kinetics analysis had shown that Km and Vmax of the higher affinity kinetic component of ATPase increased significantly by 1.2- to 1.4-fold in epileptic condition. Temperature kinetic analysis revealed 1.2-fold increase in energies of activation with decreased transition temperature. The total phospholipid (TPL) and cholesterol (CHL) contents decreased significantly with lowering of diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), phosphatidylinositol (PI), and phosphatidylserine (PS), while lysophospholipid (lyso), sphingomyelin (SPM), and phosphatidylcholine components were found to be elevated. Brain mitochondrial membrane was somewhat more fluidized in epileptic animals. Possible consequences of mitochondrial respiratory chain (MRC) dysfunction are discussed. In conclusion, impairment of MRC function along with structural alterations suggests novel pathophysiological mechanisms important for chronic epileptic condition. [Copyright &y& Elsevier]

Effects of in vivo treatment with the three antimalarials chloroquine, primaquine and quinine on rat liver mitochondrial energy transduction functions were examined. Treatment with all the three antimalarials decreased the state 3 and state 4 respiration rates drastically. The extent of inhibition was higher with pyruvate + malate as the substrate than with glutamate. The antimalarials also acted as uncouplers. The uncoupling effect was seen on site II and site III of phosphorylation; site I was unaffected. As a consequence the ADP phosphorylation rates also decreased significantly. Following antimalarials treatment the basal and Mg2 +stimulated ATPase activities increased while the DNP-stimulated ATPase activity was reduced by half. Treatment with chloroquine resulted in decreased contents of cytochromes aa3 and b; primaquine and quinine treatments increased the contents of the two cytochromes in 14 day treatment groups. [ABSTRACT FROM AUTHOR]

A simplified method for inorganic phosphate determination has been developed. The method is sensitive, easy, economic, and applicable for estimation of phosphate released in both enzymatic and nonenzymatic reactions. A mixture of hydrazine sulfate and ascorbic acid was used as the reducing agent and the conditions for the development of the molybdenum blue color were optimized. Thus in the 4.0 ml assay system, 0.4 ml of the reducing agent solution containing 20 mg each of hydrazine sulfate and ascorbic acid per milliliter of 1.0 N H2SO4 gave a rapid optimum color development with absorption maximum at 820 nm. Color development showed a linear relationship up to 10 μg Pi concentration. Thus the method has a 2.5 × higher range of Pi estimation than that of the Bartlett method. The molar extinction coefficient at 820 nm was higher than that obtained in the Bartlett procedure. Also the molybdenum blue color formed was stable up to 24 h. Under the standard assay conditions, interference from acid-labile phosphate as in the case of Na+,K+ ATPase was at the minimum. The applicability of the method for assay of microsomal Na+,K+ ATPase and glucose-6-phosphatase was checked in microassays (final volume 0.1 ml) in comparison to the conventional procedures which use 3–4 times higher volumes. Likewise the applicability of the method for phospholipid analysis was compared with that of the conventional Bartlett method. Under both test systems the results obtained by the micromethod were identical to those obtained by the conventional methods. In general the method, which rapidly produces quantitatively molybdenum blue color, not only is rapid economical, and convenient but also has wide applicability. [Copyright &y& Elsevier]

1. Effects of short-term and long-term administration of cycloheximide on rat liver mitochondrial protein synthesis have been examined and were found to be different. 2. Long-term administration of cycloheximide resulted in inhibition of total cellular protein synthesis including that of mitochondria while, at short-term intervals, 8–10% of mitochondrial protein synthesis was cycloheximide-resistant. 3. The inhibitory effect was also reflected in terms of protein synthesizing ability of mitochondria in vitro, the inhibition becoming apparent at 40 min and showing progressive increase with time. 4. The observed inhibition of mitochondrial protein synthesis by cycloheximide was not due to either inhibition of energy metabolism or alteration of amino-acid pool. 5. Cycloheximide did not enter mitochondria or sonic preparation under conditions in vitro. On the other hand, after administration of [³H]cycloheximide, significant quantities of the label were found to be associated with mitochondria and mitoribosomes. 6. These results indicated that cycloheximide reached the site of action in mitochondria under conditions in vivo but was unable to do so in vitro. 1. The results are discussed to elucidate the possible mechanisms involved in the inhibition of truly mitochondrial protein synthesis by cycloheximide. [ABSTRACT FROM AUTHOR]

Mitochondrial dysfunction is at the base of development and progression of several psychiatric and neurologic diseases with different etiologies. MtDNA/nDNA mutational damage, failure of endogenous antioxidant defenses, hormonal malfunction, altered membrane permeability, metabolic dysregulation, disruption of calcium buffering capacity and ageing have been found to be the root causes of mitochondrial dysfunction in psychatric and neurodegenerative diseases. However, the overall consequences of mitochondrial dysfunction are only limited to increase in oxidative/nitrosative stress and cellular energy crises. Thus far, extensive efforts have been made to improve mitochondrial function through specific cause-dependent antioxidant therapy. However, owing to complex genetic and interlinked causes of mitochondrial dysfunction, it has not been possible to achieve any common, unique supportive antioxidant therapeutic strategy for the treatment of psychiatric and neurologic diseases. Hence, we propose an antioxidant therapeutic strategy for management of consequences of mitochondrial dysfunction in psychiatric and neurologic diseases. It is expected that this will not only reduces oxidative stress, but also promote anaerobic energy production. © 2013 BioFactors, 39(4):393-407, 2013 [ABSTRACT FROM AUTHOR]

Abstract: Studies on Cd hepatotoxicity have focused mainly on induction of cytochrome P450 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. The substrate and temperature kinetics parameters were examined and attempts were made to seek correlation with changes in lipid/phospholipid profiles. The Na+, K+ ATPase activity decreased only in 1 week Cd-treated group but recovered at the end of 1 month. The activity resolved in two distinct kinetic components in control as well as the experimental groups. In 1 week Cd-treated group the K m value of both the components was unchanged, whereas V max value decreased. In 1-month Cd-treated group V max value only of component I increased. The catalytic efficiency of both the components was not affected in the experimental groups. In 1-week Cd-treated group the energy of activations at high-temperature range (E H) and low-temperature range (E L) decreased, whereas for 1-month Cd-treated group the energies of activations did not change. The G6Pase activity measured at 37°C was high only in 1-month Cd-treated group. The activity resolved in two kinetically distinguishable components in control as well as in the experimental groups. K m value of component I decreased in both the Cd-treated groups. In 1-month Cd-treated group the V max value of component II increased. The catalytic efficiency of G6Pase was unchanged despite changes in K m and V max. In 1-week Cd-treated group the E H and E L decreased, whereas only E L showed decrease in 1-month Cd-treated group. Cholesterol (CHL) content increased in both the Cd-treated groups. Content of lysophospholipid (Lyso), spinghomyelin (SPM) and phosphatidic acid (PA) increased, whereas phosphatidylcholine (PC) and phosphatidylserine (PS) decreased in 1-week Cd-treated group. In 1-month Cd group the Lyso, SPM, and PC increased while PC, phosphatidylethanolamine (PE) and PA decreased. In conclusion, Cd has short-term effects on microsomal Na+, K+-ATPase which are reversed by the end of 1 month and that G6Pase does not seem to be a target of Cd insult. [Copyright &y& Elsevier]

Abstract: 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. The animals were injected with a dose of 2mg of Dex/kg body weight at around 7:00 a.m. for three alternative days prior to the day of sacrifice; the control group animals received saline vehicle. We measured rates of respiration with different substrates, viz. glutamate, pyruvate+malate, succinate and ascorbate+TMPD; the contents of individual cytochromes and the dehydrogenases and ATPase activities. Dex treatment, in general, stimulated the state 3 rates of respiration rates in young animals in age-dependent and substrate-specific manner except for the 3 week group, whereas in the adults there was substantial inhibition of the respiration. The pattern of dehydrogenases activities matched with respiration rates. Dex treatment also resulted in uncoupling of the second and third site of phosphorylation in 3-week-old animals and in the adults. The contents of cytochrome aa 3, b and ATPase activities decreased significantly after Dex treatment in all the age groups. The results thus emphasize that exposure to repeated Dex treatment can significantly influence the oxidative energy metabolism of brain mitochondria in young growing animals as well as in adults. [Copyright &y& Elsevier]

Effect of thyroidectomy (Tx) and subsequent treatment with 3,5,3′-triiodo- l-thyronine (T3) or replacement therapy (TR) with T3 + l-thyroxine (T4) on the temperature kinetics properties of FoF1 adenosine triphosphatase (ATPase, ATP synthase, H+-translocating ATP synthase EC 3.6.3.14) and succinate oxidase (SO) and on the lipid/phospholipid makeup of rat kidney mitochondria were examined. Tx lowered ATPase activity, which T3 treatment restored. SO activity was unchanged in Tx but decreased further by T3 treatment. TR restored both activities. The energies of ATPase activation in the high and low temperature ranges ( E H and E L) increased in the Tx and T3 animals with decrease in phase transition temperature (Tt). TR restored E H and E L but not Tt to euthyroid levels. E H and E L of SO decreased in Tx animals. T3 and TR restored E H whereas E L was restored only in the TR group; Tt increased in both groups. Total phospholipid and cholesterol contents decreased significantly in Tx and T3-treated animals. In Tx animals, sphingomyelin (SPM) and phosphatidylcholine (PC) components decreased, while phosphatidylserine (PS) and diphosphatidylglycerol components increased. T3 and TR treatments caused decreases in SPM, phosphatidylinositol and PS. PC and phosphatidylethanolamine (PE) increased in the T3 group. TR resulted in increased lysophospolipids and PE. Changes in kinetic parameters of the two enzymes were differently correlated with specific phospholipid components. Both T3 and TR regimens were unable to restore normal membrane structure-function relationships. [ABSTRACT FROM AUTHOR]

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. Treatment with 1.0 mg DHEA resulted in increased body weights of the young rats without change in the liver weight. In the old animals the liver weight increased progressively with increasing dose of DHEA without affecting body weight. The state 3 respiration rates in liver mitochondria from old animals were, in general, lower than those in the young rats. The state 3 and state 4 respiration rates increased following DHEA treatment in dose-dependent manner bringing them close to values for young animals or beyond that with the effect being more pronounced at 1.0 mg dose. Treatment with DHEA also stimulated state 3 and state 4 respiration rates in young rats in dose-dependent manner, Contents of cytochrome aa3, b and c+c1 increased significantly in old animals in dose-dependent manner. In the young rats the lower dose (0.2 mg) of DHEA was more effective in bringing about a maximum increase in the contents of the cytochromes: the effect declined at the higher dose (1.0 mg). DHEA treatment also stimulated the mitochondrial ATPase activity in the old as well as in the young rats, The dehydrogenases activities were considerably low in the old rats compared to the values for the young animals. Treatment with DHEA stimulated dehydrogenases activities in old rats in dose-dependent manner bringing them close to values for the young animals or beyond. Treatment with lower dose (0.2 mg) of DHEA maximally stimulated dehydrogenases activities in young animals. [ABSTRACT FROM AUTHOR]

Abstract: Neonatal hypothyroidism was induced in rat pups by injecting 131I within two days of birth and the effects on kinetic properties of Na+, K+-ATPase from synaptic plasma membranes were examined. Neonatal hypothyroidism resulted in a generalized decrease in V max with ATP, Na+, K+ and Mg2+ together with an increase in the K m for ATP, appearance of a low affinity component for Na+ and allosteric characteristic for the Mg2+-dependent activity at high Mg2+ concentrations. Binding pattern for Na+ and Mg2+ changed. Our results suggest that impairment of Na+, K+-ATPase activity together with altered kinetic properties could be one of the underlying biochemical mechanism leading to central nervous system (CNS) dysfunctions as a consequence of thyroid hormone deprivation during critical stages of brain development. [Copyright &y& Elsevier]

Dexamethasone— a potent synthetic glucocorticoid— has multiple therapeutic applications and, used in all age groups, as well as for antenatal and perinatal treatments. However, side-effects of dexamethasone treatment, including those on development, are becoming increasingly apparent. Since developmental processes are energy-dependent, we examined the effects of chronic dexamethasone treatment on oxidative energy metabolism in liver mitochondria from rats belonging to different developmental age groups. Dexamethasone treatment adversely affected the state 3 respiration rates in 2- and 3-week groups and in the adults with glutamate as the substrates, whereas for pyruvate + malate, the adverse effects were seen for the 3 week rats and the adult groups. Oxidation of succinate was severely impaired in all the age groups rats. For ascorbate + TMPD as the substrate, elevated respiration was noted for the 5-week group and the impaired oxidation was observed in adults. Dexamethasone treatment also resulted in site-specific uncoupling with the effect being seen predominantly in the 3- and 5-week and adult animals. The activity of dehydrogenases decreased in a manner comparable to the respiration rates. The mitochondrial cytochromes decreased in an age-dependent manner. The ATPase activity also decreased significantly. The results thus emphasize the adverse effects of dexamethasone treatment on mitochondrial energy metabolism especially in critical age groups. [ABSTRACT FROM AUTHOR]

Effect of long-term (90-100 days) exposure of rats to soluble salt of aluminum (AlCl3) on myelin lipid profile was examined. The long-term exposure to AlCl3 resulted in a 60 % decrease in the total phospholipid (TPL) content while the cholesterol (CHL) content increased by 55 %. Consequently the TPL / CHL molar ratio decreased significantly by 62 %. The phospholipid composition of the myelin membrane changed drastically; the proportion of practically all the phospholipid classes decreased by 32 to 60 % except for phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Of the latter two, proportion of PC was unchanged while PE increased in proportion by 47 %. Quantitatively, all phospholipid classes decreased by from 42 to 76% with no change in the PE content. However the membrane fluidity was not altered in Al-treated rats. Many of the changes we observe here show striking similarities with the reported phospholipid profiles of Alzheimer brains. [ABSTRACT FROM AUTHOR]

Aluminum (Al) is considered a potential etiological factor in Alzheimer’s disease (AD). Neurotoxicity from excess brain exposure to Al is documented from both clinical observations and animal experiments. A key role of the acetylcholine system in memory disturbances that characterize AD has been reported. On this basis, we studied the effect of long-term Al feeding on kinetic properties of cholinesterases employing the rat as experimental model. Animals were given prolonged treatment with soluble salts of Al (100 mg AlCl3/kg body weight mixed with food for 100–115 days), and the kinetic properties of cholinesterases (acetylcholinesterase, AChE, and butyrylcholinesterase, BChE) were determined in different tissues. Prolonged treatment with Al had no effect on the Km values of the soluble and membrane-bound forms of AChE in the brain, but Vmax was instead decreased in all the components of soluble and membrane-bound forms of AChE in the brain. In addition, the Al treatment resulted in complete loss of the component II of erythrocyte membrane AChE. Surprisingly, after prolonged treatment with Al, higher Vmax was observed in all the components of soluble and membrane-bound forms of BChE in the heart and liver. Variable effects of Al exposure were observed on temperature kinetic properties of cholinesterases. Altogether these findings indicate that long-term Al feeding results in inhibition of AChE, while an opposite effect is observed on BChE. Decreased Vmax of the brain AChE could represent the mode of action through which Al may contribute to pathological processes in Al-induced neurotoxicity. [Copyright &y& Elsevier]

Long-term exposure to AlCl [ABSTRACT FROM AUTHOR]

Loss of erythrocyte membrane deformability is one of the most crucial factors in developing complications associated with Type II diabetes. The observed loss of erythrocyte membrane deformability could be related to structural changes in the membrane. In this context, here, we have made an attempt at gaining a better insight (quantitative as well as qualitative) into the protein and lipid contents in erythrocyte membranes and their interrelationships in Type II diabetes. Age matched control (n=12) and Type II diabetic subjects (n=22) were selected for this study. Morphological characteristics were studied by atomic force microscopy (AFM). AFM study confirmed remarkable alterations in morphology of the diabetic erythrocytes. In diabetic erythrocytes following changes were noted: (i) Significant increase in membrane as well as cytosolic proteins with a marginal increase in phospholipids content; (ii) The membrane total lipids:protein, phospholipids:protein, cholesterol:protein and phospholipids:cholesterol (mole:mole) ratios decreased significantly; (iii). A reproducible decrease in docosahexaenoic acid (DHA) and Omega-3 index with increase in Omega-6:Omega-3 ratio in membrane fatty acids; and (iv) The SDS-PAGE analysis indicated that all membrane proteins increased in almost equal proportion leading to increased membrane protein content. The observed compositional and stochiometric changes in lipids, proteins and their ratios may underlie morphological alterations and loss of deformability. [ABSTRACT FROM AUTHOR]

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. Objective: To evaluate the possible mechanism of hepatoprotective activity of withanolide-rich fraction (WRF) isolated from a methanolic extract ofWithania somniferaroots. Materials and methods: Hepatotoxicity was induced by oral administration of APAP (750 mg/kg, p.o.) for 14 d. The control group received the vehicle. APAP-treated animals were given either silymarin (25 mg/kg) or graded doses of WRF (50, 100 and 200mg/kg) 2 h prior to APAP administration. Animals were killed on 15th day and blood and liver tissue samples were collected for the further analysis. Results: In WRF-treated group, there was significant and dose-dependent (p < 0.01 andp < 0.001) decrease in serum bilirubin, ALP, AST and ALT levels with significant and dose-dependent (p < 0.01 andp < 0.001) increase in hepatic SOD, GSH and total antioxidant capacity. The level of MDA and NO decreased significantly (p < 0.01) by WRF treatment. Up-regulated mRNA expression of TNF-α, IL-1β, COX-II and iNOS was significantly down-regulated (p < 0.001) by WRF. Histological alternations induced by APAP in liver were restored to near normality by WRF pretreatment. Conclusion: WRF may exert its hepatoprotective action by alleviating inflammatory and oxido-nitrosative stress via inhibition of TNF-α, IL-1β, COX-II and iNOS. [ABSTRACT FROM AUTHOR]

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. We examined these characteristics by employing the Sino-Veda Madin-Darby canine kidney cells culture system, which under in vitro condition shows the absorption characteristics similar to the human intestinal epithelium. Thus, the aim of the present investigation was to assess the bioavailability of individual withanolides. Withanolides were diluted in Hank's buffered saline at a concentration of 2 µg/ml were tested for permeability studies carried out for 1 h duration. Permeability was measured in terms of efflux pump (Peff) in cm/s. Peff values of withanolide A (WN A), withanone (WNN), 1,2-deoxywithastramonolide (1,2 DWM), withanolide B (WN B), withanoside IV-V (WS IV-V), and withaferin A were 4.05 × 10-5, 2.06 × 10-5, 1.97 × 10-5, 1.80 × 10-5, 3.19 × 10-6, 3.03 × 10-6 and 3.30 × 10-7 respectively. In conclusion, the nonpolar and low molecular weight compounds (WN A, WNN, 1,2 DWM, and WN B) were highly permeable. As against this, the glycosylated and polar WS IV and WS V showed low permeability. Surprisingly and paradoxically, the highly biologically active withaferin A was completely impermeable, suggesting that further studies possibly using human epithelial colorectal adenocarcinoma (Caco-2) cells may be needed to delineate the absorption characteristics of withanolides, especially withaferin A. [ABSTRACT FROM AUTHOR]

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. somnifera. Therefore, we examined the effect of these deficiencies on the withanolides content of the roots. Objective: To examine the effect of molybdenum accretion in nitrogen-, phosphorus-, calcium- and potassium-deficient soils on the accumulation of withanolides in the roots of W. somnifera. Different withanolides have different therapeutic applications hence major bioactive withanolides assume importance. Materials and methods: Methanol extracts of the roots were subjected to HPTLC and individual withanolides were identified by comparing their Rf values with those of the authentic samples. Molybdenum was quantified by atomic absorption spectroscopy. Free radical scavenging activity was monitored by the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay. Results: Molybdenum content in roots of nitrogen-, phosphorus-, calcium-, potassium-deficient, and control plants were 7.02 ± 2.1, 13.1 ± 1.6, 17.1 ± 0.9, 33.5 ± 3.3, and 33.9 ± 1.6 ppm, respectively. Levels of withaferine A increased with the increase in the Mo content in roots from 7.79 ± 2.2 mg/g to 12.57 ± 3.4 mg/g. Antioxidant activity of nitrogen-deficient plants was the lowest (24.7 ± 2.2%) compared to other groups. Discussion and conclusion: It was observed that nitrogen metabolism-dependent molybdenum uptake influences the withanolides accumulation in the roots. [ABSTRACT FROM AUTHOR]

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. However, the effect was temperature-dependent. In diabetic animals the energy of activation in the low temperature range (E(L)) increased significantly while that in the high temperature range (E(H)) decreased. The latter seems to be responsible for improving catalytic efficiency in the diabetic state. Insulin treatment normalized E(H) only in the 1-month diabetic group. The phase transition temperature (Tt), decreased in diabetic animals. Insulin treatment caused an increase beyond the control value in Tt in 1-month diabetic animals. The results suggest that insulin status-dependent modulation of SO activity is a complex process. [ABSTRACT FROM AUTHOR]

Thyroid-status-dependent changes in lipid/phospholipid profiles in rat liver mitochondria were examined and attempts were made to correlate the observed changes with kinetic parameters of succinate-2,6-dichloroindophenol reductase (SDR) and membrane fluidity. Thyroidectomy caused substantial decrease in the total phospholipid and cholesterol contents; the proportion of all phospholipid components was also decreased significantly (50–100% decrease). The energy of activation, E1, for SDR decreased with a concomitant increase in the membrane fluidity. Treatment with physiologic doses or replacement doses of L-thyroxine resulted in partial restoration of lipid/phospholipid components. However, the SDR activity decreased with a simultaneous elevation in the phase-transition temperature and significant decreases in E1 and E2. Treatment with L-thyroxine restored membrane fluidity to almost euthyroid values. Regression analysis suggested a role for phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol microdomains in modulating the SDR activity. Membrane fluidity was strongly correlated with total phospholipid, cholesterol, diphosphatidylglycerol and phosphatidyl-inositol, and the ratio of cholesterol/total phospholipid (mol/mol). [ABSTRACT FROM AUTHOR]