Biochemical Effect of Dicarboxylic Acids on Oxalate Metabolism in Experimental Rats and Studies on Oxalate Degrading Bacteria

The current study is an attempt to find a means of lowering oxalate concentration in individuals susceptible to recurrent calcium oxalate stone disease.The formation of renal stone composed of calcium oxalate is a complex process that remains poorly understood and treatment of idiopathic recurrent stone formers is quite difficult and this area has attracted lots of research workers. The main objective of this work are to study the effect of certain mono and dicarboxylic acids on calcium oxalate crystal growth in vitro, isolation and characterization of oxalate degrading bacteria, study the biochemical effect of sodium glycollate and dicarboxylic acids on oxalate metabolism in experimental stone forming rats and To investigate the effect of dicarboxylic acids on oxalate metabolism in experimental hyperoxaluric rats. Oxalic acid is one of the most highly oxidized organic compound widely distributed in the diets of man and animals, and ingestion of plants that contain high concentration of oxalate may lead to intoxication. Excessive ingestion of dietary oxalate may lead to hyperoxaluria and calcium oxalate stone disease.The formation of calcium oxalate stone in the urine is dependent on the saturation level of both calcium and oxalate. Thus the management of one or both of these ions in individuals susceptible to urolithiasis appears to be important. The control of endogenous oxalate synthesis from its precursors in hyperoxaluric situation is likely to yield beneficial results and can be a useful approach in the medical management of urinary stones. A variety of compounds have been investigated to curtain endogenous oxalate synthesis which is a crucial factor, most of these compounds have not proved to be effective in the in vivo situation and some of them are not free from the toxic effect. The non-operative management of stone disease has been practiced in ancient India in the three famous indigenous systems of medicine, Ayurveda, Unani and Siddha, and proved to be effective.However the efficiency of most of these substances is still questionable and demands further study. Man as well as other mammals cannot metabolize oxalic acid. Excessive ingestion of oxalic acid can arise from oxalate rich food and from its major metabolic precursors, glycollate, glyoxylate and ascorbic acid can lead to an acute oxalate toxicity. Increasedlevels of circulating oxalate, which can result in a variety of diseases including renal failure and oxalate lithiasis. The ability to enzymatically degrade oxalate to less noxious Isubstances, formate and CO2, could benefit a great number of individuals including those afflicted with hyperoxaluria and calcium oxalate stone disease.

Kinetic Studies of Purified malate Dehydrogenase in Liver of Streptozotocindiabetic Rats and the Effect of Leaf Extract of Aegle Marmelose (L.) Correa ex Roxb

The functional basis of diabetes-mellitus to a certain extent, can be elucidated by studying diabetes-induced changes in metabolic enzymes. Malate dehydrogenase (MDH), is an enzyme directly involved in glucose metabolism. The kinetic parameters of MDH and its purified cytosolic isozyme, S-MDH, have been studied in the liver of streptozotocin- diabetic rats; also the potential of the leaf extract of A. marmelose as an was investigated. The Km of the liver enzyme increased significantly, in both crude and purified preparations in the diabetic state when compared to Lhe respective controls. Insulin as well as leaf- •extract treatment of the diabetic rats brought about a reversal of K. values to near normal. Vmax of purified S-MDH was significantly higher in the diabetic state when compared to the control. Insulin and leaf extract treatment did not reverse this change. Since MDH is an important enzyme in glucose metabolism, the variation in its quantitative and qualitative nature may contribute to the pathological status of diabetes. The fact that leaf extract of A. marmelose was found to be as effective as insulin in restoration of blood glucose and body weight to normal levels, the use of A. marmelose as potential hypoglycemic agent is suggested.

Dopamine Receptor Gene Expression In Streptozotocin Induced Diabetic Rats And Its Role In Insulin Secretion

Diabetes Mellitus is a metabolic disorder associated with insulin deficiency, which not.only affects the carbohydrate metabolism but also is associated with various central and peripheral complications. Chronic hyperglycemia during diabetes mellitus is a major initiator of diabetic microvascular complications like retinopathy, neuropathy, The central nervous system (CNS) neurotransmitters play an important role in the regulation of glucose homeostasis. These neurotransmitters mediate rapid intracellular communications not only within the central nervous system but also in the peripheral tissues. They exert their function through receptors present in both neuronal and non neuronal cell surface that trigger second messenger signaling pathways. Dopamine is a neurotransmitter that has been implicated in various central neuronal degenerative disorders like Parkinson's disease and behavioral diseases like Schizophrenia. Dopamine is synthesised from tyrosine, stored in vesicles in axon terminals and released when the neuron is depolarised. Dopamine interacts with specific membrane receptors to produce its effect. Dopamine plays an important role both centrally and peripherally. The recent identification of five dopamine receptor subtypes provides a basis for understanding dopamine's central and peripheral actions . Dopamine receptors are classified into two major groups : DA D1 like and DA D2 like. Dopamine D1 like receptors consists of DA D1 and DA D5 receptors . Dopamine D2 like receptors consists of DA D2, DA D3 and DA D4 receptors. Stimulation of the DA D1 receptor gives rise to increased production of cAMP. Dopamine D2 receptors inhibit cAMP production, but activate the inositol phosphate second messenger system . Impairment of central dopamine neurotransmission causes muscle rigidity, hormonal regulation , thought disorder and cocaine addiction. Peripheral dopamine receptors mediate changes in blood flow, glomerular filtration rate, sodium excretion and catecholamine release. The dopamine D2 receptors increased in the corpus striatum and cerebral cortex but decreased in the hypothalamus and brain stem indicating their involvement in regulating insulin secretion. Dopamine D2 receptor which has a stimulatory effecton insulin secretion decreased in the pancreatic islets during diabetes. Our in vitro studies confirmed the stimulatory role of dopamine D2 receptors in stimulation of glucose induced insulin secretion. A detailed study at the molecular level on the mechanisms involved in the role of dopamine in insulin secretion, its functional modification could lead to therapeutic interventions that will have immense clinical importance.

Production of Alkaline Protease by Free and Immobilised Cells of VIBRIO sp. Under Different Fermentation Systems and Its Application on Deproteinisation of Prawn Shell Waste for Chitin Recovery

This thesis presents a detailed account of a cost - effective approach towards enhanced production of alkaline protease at profitable levels using different fermentation designs employing cheap agro-industrial residues. It involves the optimisation of process parameters for the production of a thermostable alkaline protease by Vibrio sp. V26 under solid state, submerged and biphasic fermentations, production of the enzyme using cell immobilisation technology and the application of the crude enzyme on the deproteinisation of crustacean waste.The present investigation suggests an economic move towards Improved production of alkaline protease at gainful altitudes employing different fermentation designs utilising inexpensive agro-industrial residues. Moreover, the use of agro-industrial and other solid waste substrates for fermentation helps to provide a substitute in conserving the already dwindling global energy resources. Another alternative for accomplishing economically feasible production is by the use of immobilisation technique. This method avoids the wasteful expense of continually growing microorganisms. The high protease producing potential of the organism under study ascertains their exploitation in the utilisation and management of wastes. However, strain improvement studies for the production of high yielding variants using mutagens or by gene transfer are required before recommending them to Industries.Industries, all over the world, have made several attempts to exploit the microbial diversity of this planet. For sustainable development, it is essential to discover, develop and defend this natural prosperity. The Industrial development of any country is critically dependent on the intellectual and financial investment in this area. The need of the hour is to harness the beneficial uses of microbes for maximum utilisation of natural resources and technological yields. Owing to the multitude of applications in a variety of industrial sectors, there has always been an increasing demand for novel producers and resources of alkaline proteases as well as for innovative methods of production at a commercial altitude. This investigation forms a humble endeavour towards this perspective and bequeaths hope and inspiration for inventions to follow.

Toxicity, accumulation and depuration of heavy metalsin the brown mussel perna indica

In this thesis certain important aspects of heavy metal toxicity have been worked out. Recent studies have clearly shown that when experimental media contained more than one heavy metals, such metals could conspicuously influence the toxic reaction of the animals both in terms of quantity and nature. The experimental results available on individual metal toxicity show that, in majority of such results, unrealistically high concentrations of dissolved metals are involved. A remarkable number of factors have been shown to influence metal toxicity such as various environmental factors particularly temperature and salinity, the condition of the organism and the ability of some of the marine organisms to adapt to metallic contamination. Further, some of the more sensitive functions like embryonic and larval development, growth and fecundity, oxygen utilization and the function of various enzymes are found to be demonstrably sensitive in the presence of heavy metals. However, some of the above functions could be compensated for by adaptive process. If it is assumed that the presence of a single metal in higher concentrations could affect the life function of marine animals, more than one metal in the experimental media should manifest such effects in a greater scale. Commonly known as synergism or more than additivity, majority of heavy metals bring about synergistic reaction .The work presented in this thesis comprises lethal and sublethal toxicities of different salt forms of copper and silver on the brown mussel Perna indica. during the present investigation sublethal concentrations of copper and silver in their dent effects on survival, oxygen consumption, filtration, accumulation and depuration on Perna indica. The results are presented under different sections to make the presentation meaningful .

Biochemical & Pharmacological evaluation of liver oils of selected deep sea sharks and chimaeras of the Indian EEZ

With a seacoast of 8,1 18 km, an exclusive economic zone (EEZ) of 2 million square km, and with an area of about 30,000 square km under aquaculture, lndia produces close to six million tonnes of fish, over 4 per cent of the world fish production. While the marine waters upto 50m depth have been fully exploited, those beyond, remain unexplored. There is an ever increasing demand for fishery resources as food. The coastal fishery resources of the country are dwindling at a rapid pace and it becomes highly imperative that we search for alternate fishery resources for food. The option we have is to hunt for marine fishery resources. Studies pertaining to proximate composition, amino acid and fatty acid composition are essential to understand the nutraceutical values of these deep sea fishery resources. The present study was aimed to carry out proximate composition of deep sea fishery resources obtained during cruises onboard the FORV Sarise Sampada, to identify fishery resources which have appreciable lipid content and thereby analyse the bioactive potentials of marine lipids, to study the amino acid profile of these fishery resources, to understand the contents of SPA, MUFA and PUFA and to calculate the n3/n6 fatty acid contents. Though the presence of nutraceuticals was identified in the marine fishery resources their use as potential food resources deserve further investigation. So the study were carried out to calculate the hepatosomatic indices of sharks & chimaeras and conduct biochemical characterisation of liver oils of Apristurus indicus, Cenlrophorus scalprams, Centroselachus crepidater, Neoharriotta raleighana, and Harriotta pinnata obtained during cruises onboard the FORV Sugar Sampada.Therapeutic use of shark liver oil is evident from its use for centuries as a remedy to heal wounds and fight flu (Neil er al. 2006). Japanese seamen called it 'samedava' or "cure all". Shark liver oil is being promoted worldwide as a dietary supplement to boost the immune system, fight infections, to treat cancer and to lessen the side effects of conventional cancer treatment. These days more emphasis is laid on the nutritive benefits of shark liver oils especially on the omega 3 polyunsaturated fatty acids ( PUFAs) (Anandan er al. 2007) and alkylglycerols (AKGs) (Pugliese er al. I998) contained in them due to the high rise of inflammatory disorders such as arthritis, asthma and neurodegenerative diseases like Alzheimer’s, Parkinson’s and Schizophrenia. So the present study also evaluate the pharmacological properties with respect to analgesic, anti-inflammatory, anti pyretic and anti-ulcer effects of four different liver oils of sharks belonging to the Indian EEZ and to identify the components of oil responsible for these activities.The analgesic and anti-inflammatory activities of liver oils from Neoharriotra raleighana (NR), Centrosymnus crepidater (CC), Apristurus indicus (AI), and Centrophorus sculpratus (CS) sharks caught from the Arabian Sea and the Indian Ocean were compared. The main objectives also include determination of the cholesterol lowering effects of liver oils of Neoharriotra raleighana (NR) and Centrophorus sculpratus (CS) on the high fat diet induced dyslipidemia and to compare the impact of four isolipidemic diets, on levels of serum diagnostic marker enzymes, on lipid profile of blood and liver and antioxidant status of heart in male Albino rats. And also to study the efficacy of Centrophorus sculpratus (CS) liver oil against Complete Freund’s Adjuvant-induced arthritis and to compare the anti-inflammatory activity of this oil with a traditionally used anti-inflammatory substance gingerol (oleoresin extracted from ginger.). The results of the present study indicated that both (Centrophorus sculpratus liver oils as well as gingerol extracts proved to be effective natural remedies against CFA-induced arthritis in Albino rats.

Diversity in Structural and Spectural Characteristics of Some Transition Metal Complexes Derived From Aldehyde Based Thiosemicarbazone Ligands

The study deals with the diversity in structural and spectural characteristics of some transition metal complexes derived from aldehyde based thiosemicarbazone ligands thiosemicarbazones are a family of compounds with beneficial biological activity viz., anticancer,antitumour, antifungal, antibacterial, antimalarial, antifilarial, antiviral and anti-HIV activities. Many thiosemicarbazone ligands and their complexes have been prepared and screened for their antimicrobial activity against various types of fungi and bacteria. The results prove that the compounds exhibit antimicrobial properties and it is important to note that in some cases metal chelates show more inhibitory effects than the parent ligands. The increased lipophilicity of these complexes seems to be responsible for their enhanced biological potency. Adverse biological activities of thiosemicarbazones have been widely studied in rats and in other species. The parameters measured show that copper complexes caused considerable oxidative stress and zinc zinc complexes behaved as antioxidants. It has applications on analytical field also. Some thiosemicarbazones produce highly colored complexes with metal ions. This thesis aims to synthesis some novel thiosemicarbazone ligands and their transition metal complexes together with their physico-chemical characterization.

Effect of Pyridoxine Deficiency in Young-Rats on High-Affinity Serotonin and Dopamine Receptors

The high-affinity bindings of [3H]-5-hydroxytryptamine to serotonin S-1 receptors, [3H]-ketanserin to serotonin S-2 receptors in the cerebral cortex, [3H]- fluphenazine to dopamine D-1 receptors, and [3H]-spiroperidol to dopamine D-2 receptors in the corpus striatum were studied in pyridoxine-deficient rats and compared to pyridoxine-supplemented controls. There was a significant increase in the maximal binding (Bmax) of serotonin S-1 and S-2 receptors with a significant decrease in their binding affinities (Kd). However, there were no significant changes either in the maximal binding or binding affinity of striatal dopamine D- 1 and D-2 receptors. Receptor sensitivity seems to correlate negatively with the corresponding neurotransmitter concentrations in the pyridoxine-deficient rats.

Enhanced dopamine D2 receptor function in hypothalamus and corpus striatum: their role in liver, plasma and in vitro hepatocyte ALDH regulation in ethahol treated rats

Dopamine D2 receptors are involved in ethanol self- administration behavior and also suggested to mediate the onset and offset of ethanol drinking. In the present study, we investigated dopamine (DA) content and Dopamine D2 (DA D2) receptors in the hypothalamus and corpus striatum of ethanol treated rats and aldehyde dehydrogenase (ALDH) activity in the liver and plasma of ethanol treated rats and in vitro hepatocyte cultures. Hypothalamic and corpus striatal DA content decreased significantly (P\0.05, P\0.001 respectively) and homovanillic acid/ dopamine (HVA/DA) ratio increased significantly (P\0.001) in ethanol treated rats when compared to control. Scatchard analysis of [3H] YM-09151-2 binding to DA D2 receptors in hypothalamus showed a significant increase (P\0.001) in Bmax without any change in Kd in ethanol treated rats compared to control. The Kd of DA D2 receptors significantly decreased (P\0.05) in the corpus striatum of ethanol treated rats when compared to control. DA D2 receptor affinity in the hypothalamus and corpus striatum of control and ethanol treated rats fitted to a single site model with unity as Hill slope value. The in vitro studies on hepatocyte cultures showed that 10-5 M and 10-7 M DA can reverse the increased ALDH activity in 10% ethanol treated cells to near control level. Sulpiride, an antagonist of DA D2, reversed the effect of dopamine on 10% ethanol induced ALDH activity in hepatocytes. Our results showed a decreased dopamine concentration with enhanced DA D2 receptors in the hypothalamus and corpus striatum of ethanol treated rats. Also, increased ALDH was observed in the plasma and liver of ethanol treated rats and in vitro hepatocyte cultures with 10% ethanol as a compensatory mechanism for increased aldehyde production due to increased dopamine metabolism. A decrease in dopamine concentration in major brain regions is coupled with an increase in ALDH activity in liver and plasma, which contributes to the tendency for alcoholism. Since the administration of 10-5 M and 10-7 M DA can reverse the increased ALDH activity in ethanol treated cells to near control level, this has therapeutic application to correct ethanol addicts from addiction due to allergic reaction observed in aldehyde accumulation.

Decreased GABAA Receptors Functional Regulation 3 in the Cerebral Cortex and Brainstem of Hypoxic Neonatal Rats: 4 Effect of Glucose and Oxygen Supplementation

Hypoxia in neonates can lead to biochemical and molecular alterations mediated through changes in neurotransmitters resulting in permanent damage to brain. In this study, we evaluated the changes in the receptor status of GABAA in the cerebral cortex and brainstem of hypoxic neonatal rats and hypoxic rats supplemented with glucose and oxygen using binding assays and gene expression of GABAAa1 and GABAAc5. In the cerebral cortex and brainstem of hypoxic neonatal rats, a significant decrease in GABAA receptors was observed, which accounts for the respiratory inhibition. Hypoxic rats sup- plemented with glucose alone and with glucose and oxygen showed, respectively, a reversal of the GABAA receptors, andGABAAa1 and GABAAc5 gene expression to control. Glucose acts as an immediate energy source thereby reducing the ATP-depletion-induced increase in GABA and oxygenation, which helps in encountering anoxia. Resuscitation with oxygen alone was less effective in reversing the receptor alterations. Thus, the results of this study suggest that reduction in the GABAA receptors functional regulation during hypoxia plays an important role in mediating the brain damage. Glucose alone and glucose and oxygen supplementation to hypoxic neonatal rats helps in protecting the brain from severe hypoxic damage.

Alkaline Protease from a Non-toxigenic Vibrio sp.(V26) and its Applications

The thesis presents a detailed account of the alkaline protease produced by Vibrio sp.(V26) a mangrove isolate,and the application of this enzyme in different fields.The protease producer strain was identified on the basis of biochemical characteristice,putative virulence traits and 16S rRNA gene sequencing.The purification and characterization of the protease has been carried out. Along with this, an attempt has been made to identifiy the protease gene. The physical parameters as well as the media components influencing protease production were optimized using Response Surfce Methodology(RSM).The scale up of the application of the protease from Vibrio sp.(V26) in the dissociation of cells in animal cell culture,in the recovery of silver from used X-ray films as well as an ingredient in commercial detergents were investigated.

“Adrenergic receptors and monoamines in the brain and pancreatic islets of streptozotoein diabetic rats: Role in insulin secretion as a function of age”

I) To study the changes in the content of brain rrrorroamirres in streptozotocirr-irrduced tliabetes as a lirnction of age and to lirrd the role oliadrenal lrornroncs in diabetic state. 2) To assess the adrenergic receptor function in the brain stem ofstreptozotocin-induced diabetic rats ofdillerent ages. 3) To study the changes in the basal levels of second messenger cAMP in the brain stenr ofstreptozotocin-induced diabetic rats as a function of age. 4) To study the changes occurring in the content ofmorroamines and their metabolites in whole pancreas and isolated pancreatic islets of streptozotocin-diabetic rats as a function ofage and the effect of adrenal hormones. 5) To study the adrenergic receptors and basal levels of cAMP in isolated pancreatic islets in young and old streptozotoein-diabetic rats. 6) The in virro study of CAMP content in pancreatic islets of young and old rats and its ellect on glucose induced insulin secretion. 7) 'lhe in vitro study on the involvement of dopamine and corticosteroids in glucose induced insulin secretion in pancreatic islets as a function of age.