A comparative study of Glycogen phosphorylase from selected cepbalopods

A comparative study of Glycogen phosphorylase from selected Cephalopods is reported in this thesis. A detailed investigation of an important glycolytic enzyme, phosphorylase, from a selected species, is undertaken. Loligo vulgaris, commonly known as squid, is selected as the source for the study. Phosphorylase is the key enzyme in the mobilization of chemical energy from glycogen and its role in the regulation of carbohydrate metabolism is well established. Although a good deal of information is available about phosphorylase from terrestrial animals, not much is known about the enzyme from aquatic fauna. In order to bridge this gap and also to compare the results with the findings from other sources, phosphorylase a was isolated from this marine mollusc and its properties studied in detail in this study

Glycogen degrading enzymes and their regulation in benthic animals and estuarine fish

Alpha glucan phosphorylase plays a very significant role in glycolysis. The inhibition and activation of this enzyme have significant effect on the rate of glycolysis. The rate of glycolysis is also determined by the interconversion between the active 3 and inactive Q forms of phosphorylase by two specific enzymes called phosphorylase phosphatase and phosphorylase kinase. The allosteric properties and interconversion mechanism reported for well—studied animal muscle phosphorylases do not fall under a general pattern. Studies using purified phosphorylase from marine sources are scanty. Detailed studies using specialised tissues from more marine animals are necessary to find the factors that control the properties and activities of the enzyme. This thesis is an attempt in this direction. The thesis deals with a detailed study of the control of the phosphorylase by both allosterism and interconversion between the g and b forms from four different aquatic animals of different habitat. Phosphorylase frm the four different animal muscles were purified either partially or completely and the kinetic and control properties were studied.

Effect of leaf extract of Aegle marmelose in diabetic rats

Alloxan induced animal model was used to evaluate the potential antidiabetic effect of A . marmelose leaf extract. The diabetic animals were given insulin injection and another group A . marmelose leaf extract orally. It maintained the weight of the animals near to the control rats but a significant decrease in weight was noted it diabetic animals without any treatment . The blood elucose level in 'treated animals were near to that of control ones. Also a significantly increased glucose tolerance was observed in animals orally given the leaf extract prior to the experiment . A significant decrease in liver glycogen ( 1.24 -.07 s!/ 100 g of wet tissue) was observed in diabetic rats which was brought to almost the normal level (I.84- . 14 g; 100 g) with Icaf extract treatment. Blood urea and serum cholesterol increased ( 62.66 - 3.50 and 192.67± 13.64 mg' dl) significantly in alloxxan diabetic rats . The leaf extract treatment decreased the blood urea and serum . cholesterol (37.83 - 3.97 and 99?0±8.43 mg/dl ) to that of control ones . A similar effect was seen with insulin treatment . The results indicate that the active principle in .4. marmelose leaf extract has similar hypoglycaemic activity to insulin treatment.

Hypoglycaemic Effect of Alkaloids Preparation From Leaves of Aegle Marmelose

Alloxan induced diabetic animal model was used to evaluate the antidiabetic effect of alkaloids extracted from the leaves of Aegis marine/ose. The alkaloid extract maintained the weight of animals near to that of control ones - whereas there was a decrease in the body weight of diabetic animals. A significant increase in blood glucose (342. 14 -+- 14.89 mg/dl) was seen in diabetic animals but in alkaloid treated group the blood glucose was lowered (90: 12 +_5.81 mg/dl). There was no decrease in blood urea arid sreum cholesterol in the alkaloid treated group of diabetic animals. The liver glycogen decreased in diabetic animals (1.27+.12 g/100g of wet tissue) and the treatment brought the glycogen level to that of control ones (2.51 +.75 g/100 g of wet tissue). The result show that the alkaloid extract has hypoglycaemic activity.

Haematological Studies on the Freshwater Mussel - Lamellidens Marginalis ( Lamark, 1819 )

School of Environmental Studies, Cochin University of Science and Technology

STUDIES ON HAEMOLYMPH CONSTITUENTS OF PILA VIRENS (LAMARCK) AND THE EFFECTS OF PESTICIDES ON THE ACTIVITY PATTERN OF SELECTED ENZYMES

The present study is an attempt to understand the physiological responses of a freshwater gastropod, in terms of haematological parameters, in normal conditions as well as in various natural and man made altered conditions of the environment.Pila virens, a freshwater prosobranch,commonly found in paddy fields, ponds, and streams of Kerala is selected as the test animal for the present investigation. Various haemolymph constituents such as total carbohydrate, glycogen, total protein, total lipid, urea,ammonia,sodium,potasium, calcium, and chloride which are directly involved in the control and maintenance of different physiological systems, were analysed in the present study. Selected haematological parameters like total haemocyte number, and packed cell volume were also determined. Besides , the activity pattern of selected haemolymph enzymes such as acid phosphatase (ACP), alkaline phosphatase (ALP),Glutamate-oxaloacetate transaminase (GOT), and glutamate-pyruvate transaminase (GPT), all having diagnostic value in terms of internal defence system and metabolism of the organism, were also studied.

STUDIES ON THE BIOLOGY OF THE WEDGE CLAM DONAX INCARNATUS (GMELIN) FROM THE MALIPPURAM BEACH OF KERALA

The present investigations have considerably enhanced the existing knowledge on the biology and distribution/availability pattern of D.incarnatus in the Malippuram region. The species occurs in good concentration during October - March/April, and disappears from the area during late premonsoon and monsoon months. Recolonising the area in September, it grows fast in the subsequent months. The life span of the species is estimated to be about an year. Studies on the reproductive biology of the species have revealed that there are two spawning peaks, the major peak in February - March and minor peak, in December. The salinity regime of the area influences the reproductive activity. These observations form the original contribution in the thesis. The information on variation in water content, protein,glycogen and lipid levels in relation to reproductive cycle has helped to a better understanding of the gametogenic activity and spawning of the species. Similarly, the findings on salinity tolerance and filtration rate have shown that small sized clams exhibit greater tolerance range than larger clams, and grow at a faster rate with active metabolism. It is hoped that these information would considerably add to the present knowledge of the basic facts which are relevant to the improvement and management of the clam fishery of this region.

Toxicological effects of Copper and Mercury on the fish Macrones gulio (Hamilton-Buchanan)

Asha M. R This thesis Entitled Toxicological effects of copper and mercury on the fish macerones gulio (hamiloton – buchanan).Chapter 1. In this chapter, a broad outline of heavy metal uptake, requirement of a suitable bio — monitoring organism, criteria for a standard test fish, and suitability of Macrones gulig for the toxicological study are given. Chapter 2. This chapter deals with the lethal toxicity bioassays to find the 96 hr LC 50 of copper and mercury for the fish Macrones gglig. The experimental results indicated that of the two metals tested, copper was more toxic than mercury.Chapter 3. The effect of copper and mercury on the haemoglobin, haematocrit, erythrocyte count, MCV, MCH and MCHC was studied.Chapter 4. The glycogen and protein contents of liver and muscle after exposure to copper and mercury were studied. There was a significant decrease of glycogen in the liver and muscle of metal treated fishes.Chapter 5. The histopathological changes of the tissues like liver, kidney and gill after exposure to copper and mercury were studied.

Bio-chemical changes associated with processing of shell fishes and flavour constituents of body meat and claw meat'of crab

In this thesis all these aspects are taken into consideration. Extensive studies were conducted on all aspects of processing of crabs, mussels and clams. The species taken for studies are commercially used ones namely Scylla sereta, perna viridis, and villorita cyprinoids. In Chapter 4.1 with regard to crab) the following aspects on their handling and processing are reported seasonal variation of chemical constituents, changes taking place during ice storage, freezing, canning etc. In Chapter 4._2 with regard to mussel, the relation between age (size) and chemical constituents, changes taking place during ice storage, freezing, canning etc. are reported and in Chapter 4.3 the changes taking place in clam muscle during icing and freezing are reported and the ame rebility of ice stored clams for canning purpose is reported.The interference of high concentration of glycogen in mussel and clam muscles during the colour development of ribose (Me-jbaum's method) is observed and remedial step are taken to minimise the interference.

Studies On Some Aspects Of Biology Of Two Estuarine Fishes Megalops Cyprinoides And Scatophagus Argus

Based on the adaption of fishes to their habitat, they are divided into three ecological groups - marine, fresh water and estuarine or brackish water forms. Estuarine fishes inhabit the less saline region of the sea, estuaries and other inland waters. These fishes are more subjected to pollution than fresh water fishes or marine fishes as they encounter pollutants present in the outgoing river water and the incoming sea water during low and high tides respectively. So, the study of the biology of the estuarine fishes has become unavoidable to assess their suitability in aquaculture. The development of both capture and culture fisheries related to any brackish water system is dependent on the availability of scientific‘ data on the various biological factors in respect of the different species. Such a study on fishes will be helpful in formulating suitable schemes for the management of brackish water for capture and culture fisheries. It was therefore felt that a study of the biological and biochemical aspects of two estuarine fishes Megalops cyprinoides Broussonet and Scatophargus Bloch which are not fully exploited in aquaculture programmes, was worth undertaking. The present study is expected to advance our knowledge on the biology of the two fishes which are very desirable for brackish water fish farming

Biochemical and Biotechnological Studies on the Cyanobacterium Synechocystis salina Wislouch

The group cyanobacteria includes a large number of organisms characterised by a low state of cellular organization. Their cells lack a well defined nucleus. Cell division is by division of the protoplast by an ingrowth of the septum. These organisms are characterised generally by a blue green colouration of the cell, the chief pigments being chlorophyll-a, carotenes, xanthophylls, C phycocyanin and C phycoerythrin. The product of photosynthesis is glycogen. These organisms lack flagellate reproductive bodies and there is a total lack of sexual reproduction. They are also unique because of the presence of murein in the place of cellulose (cell wall) and the absence of chloroplast, mitochondria and endoplasmic reticulum. Just like bacteria some of them possess Plasmids and can fix atmospheric nitrogen. In the present study growth kinetics, heavy metal tolerance, tolerance mechanisms, heavy metal intake, and antibacterial activity of §ynechocystics salina Wislouch - a nanoplanktonic, euryhaline, Cyanobacterium present in Cochin back waters has been carried out for the potential biotechnological application of this organism. _§; salina occur as small spherical cells of 3n diameter (sometimes in pairs) with bluish green colour. The species is characterised by jerky movement of the cells and is structrually similar to other cyanobacteria

Studies on the enzyme α-glucan phosphorylase

ac-qlncnn phosphorylsse is en ilportent enzyme in glycoiysis. It is the first used knows to exhibit ellosteric properties and lance its inhibition end ectivetion have significant effect on the rete ot qlycolysis. The thesis deals with 11 detailed study of the structure. inhibition and control or this snlrlls from rabbit uncle and troll e merino eninelo ‘the thesis is divided into two parts. Port 1 deals with studies on rabbit uncle glycogen phospherylese. After e review of the relevant literetnre (Chapter 1) the inhibition and chancel sodiiicetion studies on rabbit ensyle ere discussed in chepters 2 to 5. Chapter 6. gives the methods used for the study

studies on the clam meretrix casta(chemnitz)off cochin barmouth.

It is the object of the present study to contribute so much information as possible on the biology and economy of M.casta on the south west coast of India. It includes investigations on the growth of the species in three dimensions.

Investigation on key molecular targets responsible for antidiabetic properties of Symplocos cochinchinensis (Lour.) S. Moore

Diabetes mellitus is a heterogeneous metabolic disorder characterized by hyperglycemia with disturbances in carbohydrate, protein and lipid metabolism resulting from defects in insulin secretion, insulin action or both. Currently there are 387 million people with diabetes worldwide and is expected to affect 592 million people by 2035. Insulin resistance in peripheral tissues and pancreatic beta cell dysfunction are the major challenges in the pathophysiology of diabetes. Diabetic secondary complications (like liver cirrhosis, retinopathy, microvascular and macrovascular complications) arise from persistent hyperglycemia and dyslipidemia can be disabling or even life threatening. Current medications are effective for control and management of hyperglycemia but undesirable effects, inefficiency against secondary complications and high cost are still serious issues in the present prognosis of this disorder. Hence the search for more effective and safer therapeutic agents of natural origin has been found to be highly demanding and attract attention in the present drug discovery research. The data available from Ayurveda on various medicinal plants for treatment of diabetes can efficiently yield potential new lead as antidiabetic agents. For wider acceptability and popularity of herbal remedies available in Ayurveda scientific validation by the elucidation of mechanism of action is very much essential. Modern biological techniques are available now to elucidate the biochemical basis of the effectiveness of these medicinal plants. Keeping this idea the research programme under this thesis has been planned to evaluate the molecular mechanism responsible for the antidiabetic property of Symplocos cochinchinensis, the main ingredient of Nishakathakadi Kashayam, a wellknown Ayurvedic antidiabetic preparation. A general introduction of diabetes, its pathophysiology, secondary complications and current treatment options, innovative solutions based on phytomedicine etc has been described in Chapter 1. The effect of Symplocos cochinchinensis (SC), on various in vitro biochemical targets relevant to diabetes is depicted in Chapter 2 including the preparation of plant extract. Since diabetes is a multifactorial disease, ethanolic extract of the bark of SC (SCE) and its fractions (hexane, dichloromethane, ethyl acetate and 90 % ethanol) were evaluated by in vitro methods against multiple targets such as control of postprandial hyperglycemia, insulin resistance, oxidative stress, pancreatic beta cell proliferation, inhibition of protein glycation, protein tyrosine phosphatase-1B (PTP-1B) and dipeptidyl peptidase-IV (DPPxxi IV). Among the extracts, SCE exhibited comparatively better activity like alpha glucosidase inhibition, insulin dependent glucose uptake (3 fold increase) in L6 myotubes, pancreatic beta cell regeneration in RIN-m5F and reduced triglyceride accumulation in 3T3-L1 cells, protection from hyperglycemia induced generation of reactive oxygen species in HepG2 cells with moderate antiglycation and PTP-1B inhibition. Chemical characterization by HPLC revealed the superiority of SCE over other extracts due to presence of bioactives (beta-sitosterol, phloretin 2’glucoside, oleanolic acid) in addition to minerals like magnesium, calcium, potassium, sodium, zinc and manganese. So SCE has been subjected to oral sucrose tolerance test (OGTT) to evaluate its antihyperglycemic property in mild diabetic and diabetic animal models. SCE showed significant antihyperglycemic activity in in vivo diabetic models. Chapter 3 highlights the beneficial effects of hydroethanol extract of Symplocos cochinchinensis (SCE) against hyperglycemia associated secondary complications in streptozotocin (60 mg/kg body weight) induced diabetic rat model. Proper sanction had been obtained for all the animal experiments from CSIR-CDRI institutional animal ethics committee. The experimental groups consist of normal control (NC), N + SCE 500 mg/kg bwd, diabetic control (DC), D + metformin 100 mg/kg bwd, D + SCE 250 and D + SCE 500. SCEs and metformin were administered daily for 21 days and sacrificed on day 22. Oral glucose tolerance test, plasma insulin, % HbA1c, urea, creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin, total protein etc. were analysed. Aldose reductase (AR) activity in the eye lens was also checked. On day 21, DC rats showed significantly abnormal glucose response, HOMA-IR, % HbA1c, decreased activity of antioxidant enzymes and GSH, elevated AR activity, hepatic and renal oxidative stress markers compared to NC. DC rats also exhibited increased level of plasma urea and creatinine. Treatment with SCE protected from the deleterious alterations of biochemical parameters in a dose dependent manner including histopathological alterations in pancreas. SCE 500 exhibited significant glucose lowering effect and decreased HOMA-IR, % HbA1c, lens AR activity, and hepatic, renal oxidative stress and function markers compared to DC group. Considerable amount of liver and muscle glycogen was replenished by SCE treatment in diabetic animals. Although metformin showed better effect, the activity of SCE was very much comparable with this drug. xxii The possible molecular mechanism behind the protective property of S. cochinchinensis against the insulin resistance in peripheral tissue as well as dyslipidemia in in vivo high fructose saturated fat diet model is described in Chapter 4. Initially animal were fed a high fructose saturated fat (HFS) diet for a period of 8 weeks to develop insulin resistance and dyslipidemia. The normal diet control (ND), ND + SCE 500 mg/kg bwd, high fructose saturated fat diet control (HFS), HFS + metformin 100 mg/kg bwd, HFS + SCE 250 and HFS + SCE 500 were the experimental groups. SCEs and metformin were administered daily for the next 3 weeks and sacrificed at the end of 11th week. At the end of week 11, HFS rats showed significantly abnormal glucose and insulin tolerance, HOMA-IR, % HbA1c, adiponectin, lipid profile, liver glycolytic and gluconeogenic enzyme activities, liver and muscle triglyceride accumulation compared to ND. HFS rats also exhibited increased level of plasma inflammatory cytokines, upregulated mRNA level of gluconeogenic and lipogenic genes in liver. HFS exhibited the increased expression of GLUT-2 in liver and decreased expression of GLUT-4 in muscle and adipose. SCE treatment also preserved the architecture of pancreas, liver, and kidney tissues. Treatment with SCE reversed the alterations of biochemical parameters, improved insulin sensitivity by modifying gene expression in liver, muscle and adipose tissues. Overall results suggest that SC mediates the antidiabetic activity mainly via alpha glucosidase inhibition, improved insulin sensitivity, with antiglycation and antioxidant activities.