Biswaranjan Paital

Antioxidant defenses and oxidative stress parameters in tissues of mud crab (Scylla serrata) with reference to changing salinity

The effects of salinity (10, 17 and 35 ppt) on O(2) consumption, CO(2) release and NH(3) excretion by crabs and oxidative stress parameters and antioxidant defenses of its tissues were reported. An increase in salinity caused a decrease in O(2) consumption and CO(2) release and an increase in ammonia excretion by crabs. Lipid peroxidation, protein carbonyl, H(2)O(2) levels and total antioxidant capacity of the tissues elevated significantly at 35 ppt salinity except in abdominal muscle where H(2)O(2) content was low. Ascorbic acid content of tissues was higher at 17 ppt salinity than at 10 and 35 ppt salinities. With increasing salinity, a gradual decrease in SOD, an increase in catalase, no change in GPx and a decrease followed by an increase in GR activities were recorded for abdominal muscle. While for hepatopancreas, an increase followed by a decrease in SOD and catalase, decrease in GPx and GR activities were noticed with increasing salinity. In the case of gills, a decrease followed by an increase in SOD, a decrease in catalase and GPx and an increase in GR activities were noted when the salinity increased from 10 ppt to 35 ppt. These results suggest that salinity modulation of oxidative stress and antioxidant defenses in Scylla serrata is tissue specific.

Alleviation of enhanced oxidative stress and oxygen consumption of L-thyroxine induced hyperthyroid rat liver mitochondria by vitamin E and curcumin.

In the present study, the role of vitamin E and curcumin on hyperthyroidism induced mitochondrial oxygen consumption and oxidative damage to lipids and proteins of rat liver are reported. Adult male rats were rendered hyperthyroid by administration of 0.0012% l-thyroxine in their drinking water, while vitamin E (200 mg/kg body weight) and curcumin (30 mg/kg body weight) were supplemented orally for 30 days. Hyperthyroidism induced elevation in serum aspartate aminotransferase and alanine aminotransferase activities were reduced significantly in response to vitamin E and curcumin treatment. On the other hand, effects of vitamin E and curcumin on hyperthyroidism induced hepatic complexes I and II mediated respiration were found to be different. While curcumin administration ameliorates hyperthyroidism induced state 3 and state 4 respiration in complex I, vitamin E treatment was effective only in reducing state 4 respiration of complex I. On the contrary, curcumin administration was ineffective in modulating hyperthyroidism induced complex II respiration, but vitamin E treatment to hyperthyroid rats resulted in augmentation of complex II respiration both at state 3 and state 4 level. Moreover, vitamin E and curcumin treatment resulted in alleviation of hyperthyroidism induced lipid peroxidation. Enhanced protein carbonylation in hyperthyroid rats is decreased only in response to simultaneous supplementation of vitamin E and curcumin. Above findings suggest that both vitamin E and curcumin have differential regulation on complexes I and II mediated mitochondrial respiration and have a protective role against L-thyroxine induced hepatic dysfunction and oxidative stress.

Supplementation of curcumin and vitamin E enhances oxidative stress, but restores hepatic histoarchitecture in hypothyroid rats.

AIMS In the present study, the effects of vitamin E and curcumin on hepatic dysfunction, mitochondrial oxygen consumption as well as hyperlipidemia in hypothyroid rats are reported. MAIN METHODS Adult male rats were rendered hypothyroid by administration of 0.05% 6-n-propyl-2-thiouracil (PTU) in their drinking water, while vitamin E (200 mg/kg body weight) and curcumin (30 mg/kg body weight) were supplemented orally for 30 days. KEY FINDINGS Hypothyroidism-induced elevation in serum aspartate aminotransferase activity was found to decline in vitamin E and curcumin treated rats. Nevertheless, distorted histoarchitecture revealed in hypothyroid rat liver was alleviated to normal by vitamin E and curcumin treatment. Regulation of hypothyroidism induced decrease in complexes I and II mediated mitochondrial respiration by vitamin E and curcumin was found to be different. Administration of curcumin to hypothyroid rats alleviates the decreased state 4 respiration and increased respiratory control ratio (RCR) level in complex I mediated mitochondrial oxygen consumption, whereas complex II mediated respiration was not influenced by exogenous antioxidants. Although, increase in serum concentration of total cholesterol was not modified by exogenous antioxidants, increased level of non-high-density lipoprotein cholesterol (non-HDL-C) in serum of hypothyroid rats was further enhanced by vitamin E and curcumin. Moreover, a significant elevation in mitochondrial lipid peroxidation and protein carbonylation was noticed in hypothyroid groups treated with vitamin E and curcumin. SIGNIFICANCE The present study suggests that supplementation of curcumin and vitamin E enhances oxidative stress parameters and hyperlipidemia; nevertheless, it protects hypothyroid-induced altered rectal temperature, serum transaminase activity and hepatic histoarchitecture.

Effects of salinity on O2 consumption, ROS generation and oxidative stress status of gill mitochondria of the mud crab Scylla serrata

Mitochondrial respiration, activities of electron transport chain enzymes and formation of oxidative stress parameters were investigated in mitochondria isolated from gill tissue of mud crabs (Scylla serrata) as a function of salinity (10 ppt, 17 ppt and 35 ppt). Mitochondrial oxygen consumption rate was higher for succinate as substrate compared with those of glutamate, malate and pyruvate. Complex I and complex II mediated respirations were higher at low salinity (10 ppt) than high salinity (17 ppt and 35 ppt). Although activities of electron transport chain enzymes particularly complexes I (EC 1.6.5.3), II (EC 1.3.99.1) and II-III (EC 1.3.2.1) were elevated linearly in response to salinity treatment, activity of complex V (ATPase, EC 3.6.1.34) was decreased at 35 ppt salinity. However, ATPase activity was higher at 17 ppt salinity in comparison to 10 ppt and 17 ppt salinity. Results of the experiment suggest that high salinity (35 ppt) causes hypoxic state in mitochondria of mud crabs. Hypoxic condition induced by high salinity was accompanied with increased hydrogen peroxide production resulting oxidative stress in mitochondria of crabs. A possible mechanism of hypoxia-induced reactive oxygen species generation and OS due to salinity stress in the crabs is discussed.

Seasonal variability of antioxidant biomarkers in mud crabs (Scylla serrata).

Studies on oxidative stress (OS) in crustacea are widely used as ecotoxicological indices to assess the environment risk produced by the impact of several stressor and pollutants. In the present study, effects of seasonality on OS physiology markers such as antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase), small antioxidant molecules (ascorbic acid and reduced glutathione), oxidative stress indices (lipid peroxidation, protein carbonylation and hydrogen peroxide) and total antioxidant capacity in hepatopancreas, gills and abdominal muscle of adult mud crab Scylla serrata, sampled from Chilika lagoon of India, were determined in winter, summer and rainy seasons. Results indicate that variations in enzymatic and non-enzymatic antioxidants with relation to season were not only tissue specific but also were gender specific. The levels of OS parameters were higher in hepatopancreas in comparison to gills and abdominal muscle of the crabs in all seasons. OS indices in tissues of the crabs were mainly higher in summer season when temperature and salinity of the lagoon were high with low oxygen content. Although OS was lower in winter season and moderate in rainy season in tissues of male crabs, it was higher in gills and hepatopancreas of females in rainy season. Correlation analyses between hydrological parameters of the lagoon (temperature, salinity and dissolved oxygen content) and OS physiology parameters in tissues of crabs suggest that abiotic factors influence the levels of antioxidant enzymes and, thereby the OS status in a tissue and sex specific manner. Collectively, the results of the present work suggest that further investigation is warranted before using OS parameters in S. serrata as biomarkers to monitor estuarine environment as these are influenced by gender, tissue and season.

An Overview of Seasonal Changes in Oxidative Stress and Antioxidant Defence Parameters in Some Invertebrate and Vertebrate Species

Antioxidant defence system, a highly conserved biochemical mechanism, protects organisms from harmful effects of reactive oxygen species (ROS), a by-product of metabolism. Both invertebrates and vertebrates are unable to modify environmental physical factors such as photoperiod, temperature, salinity, humidity, oxygen content, and food availability as per their requirement. Therefore, they have evolved mechanisms to modulate their metabolic pathways to cope their physiology with changing environmental challenges for survival. Antioxidant defences are one of such biochemical mechanisms. At low concentration, ROS regulates several physiological processes, whereas at higher concentration they are toxic to organisms because they impair cellular functions by oxidizing biomolecules. Seasonal changes in antioxidant defences make species able to maintain their correct ROS titre to take various physiological functions such as hibernation, aestivation, migration, and reproduction against changing environmental physical parameters. In this paper, we have compiled information available in the literature on seasonal variation in antioxidant defence system in various species of invertebrates and vertebrates. The primary objective was to understand the relationship between varied biological phenomena seen in different animal species and conserved antioxidant defence system with respect to seasons.

In silico Prediction and Characterization of 3D Structure and Binding Properties of Catalase from the Commercially Important Crab, Scylla serrata

The enzyme catalase breaks down H2O2, a potentially harmful oxidant, to H2O and O2. Besides oxidase activity, the enzyme also exhibits peroxidase activity. Therefore, it plays an important role in maintaining health and regulating pathophysiology of the organisms. However, 3D structure of this important enzyme in invertebrates particularly in crabs is not yet available. Therefore, an attempt has been made to predict the structure of the crab catalase and to envisage its catalytic interaction with H2O2. A three dimensional model of crab catalase was constructed using the NADPH binding site on Beef Liver catalase from Bos taurus (PDBID: 7CAT) as template by comparative modeling approach. Backbone conformation of the modeled structure by PROCHECK revealed that more than 98% of the residues fell in the allowed regions, ERRAT results confirmed good quality of modeled structure and VERIFY3D profile was satisfying. Molecular docking has been used to know the binding modes of hydrogen peroxide with the crab catalase protein. The receptor structures used for docking were derived from molecular dynamics (MD) simulations of homology modeled structure. The docking results showed that the three important determinant residues Arg68, Val70 and Arg108 in catalase were binding with H2O2 as they had strong hydrogen bonding contacts with the substrate. Our analysis provides insight into the structural properties of crab catalase and defines its active sites for binding with substrate. These data are important for further studies of catalase of invertebrates in general and that of crabs in particular.

Effects of temperature on complexes I and II mediated respiration, ROS generation and oxidative stress status in isolated gill mitochondria of the mud crab Scylla serrata.

Effects of fluctuations in habitat temperature (18-30°) on mitochondrial respiratory behavior and oxidative metabolic responses in the euryhaline ectotherm Scylla serrata are not fully understood. In the present study, effects of different temperatures ranging from 12 to 40°C on glutamate and succinate mediated mitochondrial respiration, respiratory control ratio (RCR), ATP generation rate, ratio for the utilization of phosphate molecules per atomic oxygen consumption (P/O), levels of lipid peroxidation and H2O2 in isolated gill mitochondria of S. serrata are reported. The pattern of variation in the studied parameters was similar for the two substrates at different temperatures. The values recorded for RCR (≥3) and P/O ratio (1.4-2.7) at the temperature range of 15-25°C were within the normal range reported for other animals (3-10 for RCR and 1.5-3 for P/O). Values for P/O ratio, ATP generation rate and RCR were highest at 18°C when compared to the other assay temperatures. However, at low and high extreme temperatures, i.e. at 12 and 40°C, states III and IV respiration rates were not clearly distinguishable from each other indicating that mitochondria were completely uncoupled. Positive correlations were noticed between temperature and the levels of both lipid peroxidation and H2O2. It is inferred that fluctuations on either side of ambient habitat temperature may adversely influence mitochondrial respiration and oxidative metabolism in S. serrata. The results provide baseline data to understand the impacts of acute changes in temperature on ectotherms inhabiting estuarine or marine environments.

Longevity of animals under reactive oxygen species stress and disease susceptibility due to global warming.

The world is projected to experience an approximate doubling of atmospheric CO2 concentration in the next decades. Rise in atmospheric CO2 level as one of the most important reasons is expected to contribute to raise the mean global temperature 1.4 °C-5.8 °C by that time. A survey from 128 countries speculates that global warming is primarily due to increase in atmospheric CO2 level that is produced mainly by anthropogenic activities. Exposure of animals to high environmental temperatures is mostly accompanied by unwanted acceleration of certain biochemical pathways in their cells. One of such examples is augmentation in generation of reactive oxygen species (ROS) and subsequent increase in oxidation of lipids, proteins and nucleic acids by ROS. Increase in oxidation of biomolecules leads to a state called as oxidative stress (OS). Finally, the increase in OS condition induces abnormality in physiology of animals under elevated temperature. Exposure of animals to rise in habitat temperature is found to boost the metabolism of animals and a very strong and positive correlation exists between metabolism and levels of ROS and OS. Continuous induction of OS is negatively correlated with survivability and longevity and positively correlated with ageing in animals. Thus, it can be predicted that continuous exposure of animals to acute or gradual rise in habitat temperature due to global warming may induce OS, reduced survivability and longevity in animals in general and poikilotherms in particular. A positive correlation between metabolism and temperature in general and altered O2 consumption at elevated temperature in particular could also increase the risk of experiencing OS in homeotherms. Effects of global warming on longevity of animals through increased risk of protein misfolding and disease susceptibility due to OS as the cause or effects or both also cannot be ignored. Therefore, understanding the physiological impacts of global warming in relation to longevity of animals will become very crucial challenge to biologists of the present millennium.

In silico prediction of 3D structure of Mn superoxide dismutase of Scylla serrata and its binding properties with inhibitors

In the present study, we used computational methods to model crab and rat MnSOD using the crystal structure of MnSOD from Homo sapiens (PDB code: 1MSD) as template by comparative modeling approach. We performed molecular dynamics simulations to study dynamic behavior of the crab MnSOD. The modeled proteins were validated and subjected to molecular docking analyses. Molecular docking tool was used to elucidate a comparative binding mode of the crab and rat SOD with potent inhibitors of SOD such as hydrogen peroxide (H2O2), potassium cyanide (KCN) and sodium dodecyl sulphate (SDS). The predicted valid structure of crab MnSOD did not show any interaction with KCN but close interaction with H2O2 and SDS. A possible inhibitory mechanism of SDS and H2O2 due to their interaction with the amino acids present in the active site of the MnSOD of the above two animals are elucidated. This allowed us to predict the binding modes of the proteins to elucidate probable mode of action and sites of interference.