Study was conducted to investigate the aeroallergens of Imphal with special reference to respiratory allergy. The flora of Imphal was never free from flowering throughout the period of investigation. These plants were catagorised into four groups viz. trees, shrubs and undershrubs, climbers and herbs (wild and cultivated). Out of 257 species of wild and cultivated angiospermic plants collected, trees contributed 57 species, shrubs and undershrubs 68 species, herbs 115 species and climbers contributed 17 species. Plants like Hibiscus rosa sinensis, Acacia nilotica, Callistemon linearis, Canna indica, Rosa spp, solanum melongena, Lantana camera, Duranta plumeri, Bougainvillea specatbilis, etc. were found flowering almost round the year. From the allergenic point of view, the following plants are common in Imphal and its suburbs-tress (Melia azadirachta, Mangifera indica, Cassia fistula, Callistemen linearis, Eucalyptus globosus, Carica papaya etc.) shrubs and undershrubs (Rosa indica, Datura metal, Lantana camera, Ricinus communis. Climbers - Cucurbita maxima, Lathyrus sativus, etc.) and herbs (Ageratum conyzoids, Amaranthus spinosa, Artemisia vulgaris, Chenopo- dium album , C.ambrisoides, Coriandrum sativum, Cannabis sativa, Cyperus rotundus, Cynodon dactylon, Erogrostis gangetica, Oryza sativa, Poa annua, Setaria glauca, Targetes erecta, Plantago major, Xanthium strumarium, etc) In Imphal the pollen content of the atmosphere was found to be maximum from July to August, minimum from April to May and moderate from November to January. On the basis of the atmospheric pollen incidence four principal pollen seasons in Imphal could be identified. Trees predominate from February to April, shrubs and undershrubs from December to February, grasses from July to September and weeds from August to October. Among the pollen types identified from the air of Imphal, Poaceae (Cynodon, Digitaria, Eleusine, Oryza, Panicum, Setaria, Imperata and Zea), Cyperaceae (Cyperus), Alnus, Callistemon, Artemisia, Eucalyptus, Carica, Syzygium, Azadirachta, Coriandrum, Morus, Xanthium, Albizzia, Amaranthus, Ageratum, Lantana, Mangifera, Ricinus, Cassia, Rumex, Bauhinia, Brassica, Chenopodium and Acacia, were proved allergenically significant in the published literature. In this region allergenic pollens of weeds which cause symptoms of rhinitis or asthma, were found to be most important followed by those of grasses and trees. Aspergilli-Penicilli ranked second in total yearly contribution. The maximum concentration of Aspergilli-Penicilli spores were recorded during March and October 1989. In the studies where culture plate method was employed to study outdoor air-spores, Aspergillus spp. have been found to contribute to a significant proportion of the total colonies. Six Aspergillus species were identified. Aspergillus niger ranked third in yearly total population of the four Alternaria species identified Alternaria . Alternata ranked fourth in their contribution to yearly total population. Mucor globosus was recorded throughout the investigation period except in June 1988 but was found to be of less significance. Of the four Cladosporium species identified, Cladosporium cladosporioides occupied first place while C. herbarium occupied second place in their yearly total population. Seasonal periodicities of 20 allergenic fungal spores and 15 aller- genic fungal species as compared with meteorological parameters for two years were recorded. In general continuous rain washed off airborne spores. The number of dermatophagoides (Dermatophagoides farinae and D. pteronyssinus) in house dust showed a periodical increase and decrease. Maximum numbers were recorded in August, September , October, February, March and April. The peak growth of two mite species took place in August-September while minimum in the months of January to April. The role of microorganisms (fungi and bacteria) found in the nasal swab of the patients attending ENT OPD revealed no correlation between the nasal swab microflora of patients and respiratory allergy. However, record of allergic case history among the patients attending ENT OPD and Chest and TB Clinics of Regional Medical College Hospital, Lamphel (Imphal, Manipur) and clinical examination of patients gave some clues for diagnosis of respiratory allergy.

A survey of air borne pollen grains, fungal spores and other bio- components in relation to the incidence of allergenic diseases at Gwalior (M.P.) was undertaken from April 1988 to March 1991. The study revealed that about 310 angiospermic plant species are dominant in Gwalior. The pollination periods of these species vary from season to season; maximum number of trees and shrubs exhibit flowering during February-March, and herbs and grasses in August-September. A total of 67 pollen and 35 spore types were observed in the air in the year 1989, whereas in 1990 a total of 61 pollen and 23 spore types were recorded. A good number pollen and spore types were common during both the years. Occurrence of these biocomponents was directly related with the flowering of plants, climatic factors and other sources. Pollen of certain taxa such as Asteracease, Malvaceae, Amaranthaceae, Chenopodia- ceae, Poaceae etc. were found to be present in the air throughout the year. A total of 155 patients suffering from various allergic disorders were diagnosed and intradermal skin sensitivity test with 92 allergens made. Maximum allergic attacks were recorded during winter and summer seasons. Perennial disorders were commoner than seasonal. Most of the allergic patients did not have allergic family history. Allergic incidences were common among housewives followed by students, labourers, social workers, etc. More attacks were observed in the patients in the 31-40 years age group. Patients exhibited more sensitivity to insect allergens followed by pollen, fungus, dust and others. Smokers exhibited more allergic incidences than non-smokers. The mode of diet, i.e. vegetarian and non-vegetarian did not have effect on allergic manifestations. During the dominance of insects and other biocomponents in the air, the allergic disorders caused by them were also prevalent among patients. Pollen grains of anemophilous plant species are directly concerned with flowering of plants and climatic factors.

Study was carried out to observe the effects of an integrated approach of yoga therapy on atopic bronchial asthma patients, when compared to a group of control subjects, who were followed up at the same intervals, but who did not practice yoga. There were 67 patients in the `yoga' group, however, some of the data were not available for the whole group. On the whole, at various stages of the follow up there was a suggestion of improvement in terms of a significant reduction in their medication, increase in peak flow rate, in breath holding time, and in chest expansion. The FEV1/FVC percentage. ratio. The indices of bronchial lability changed as follows precent rise in PER decreased (indicating improvement), percent fall in PFR showed changes in both directions, and the sum of the two showed an increase. The eosinophils (per 100 cells) and the absolute eosinophil counts showed a general trend of reduction. The skin prick test results showed that at each stage of the follow up the number of patients showing improvement (reduced response to allergens) was more than those showing worsening. Out of a total of 23 patients for whom the serum IgE data were available, 14 showed no change, four showed a decrease and five showed an increase. Among the six control group patients, the small sample size makes it difficult to make any definite statements. There was a significant reduction in medication, an increase in PFR, and in breath holding time. The FEV1/FVC percent ratio, as well as eosinophil counts did not show a definite change. The indices of bronchial lability (% rise, % fall) showed a trend of reduction. The skin prick test results showed that at each stage the number of patients who improved were more than those who worsened. Serum IgE levels tended to increase more often than decrease or showed no change. In conclusion, it can be said that the improvement among the 'yoga' group was more apparent than for the 'control' group, with respect to the number of eosinophils per 100 cells, the absolute eosinophil count, and serum IgE levels. However, it must be emphasized that these conclusions are preliminary. Further studies on large number of control group patients are essential.

Aims of the investigation included study of the entire anatomical de- rangement of the patients suffering from exstrophy complex by using clinical as well as other investigative tools including surgical dissection, study of the histological status of the abnormally developed tissues and organs with special reference to detrusor and bladder neck region, study of human embryos by dissections and various investigative tools with special reference to genito urinary system and related systems which are affected in Exstrophy complex, assessment of the accuracy of the existing theories of the genesis, correlatation of the observations made during the study and develop a hypothesis of genesis of exstrophic malformations and development of better surgical treatment of exstrophic malformations so as to correct the total problem with minimum number of operations. The anatomical observations seem to indicate that the theories of genesis are grossly inadequate. It appears from the indirect evidence that the embryogenesis is likely to be very different than so far assumed. A number of variations are noted in the anatomical features of the conventional groups of esxtrophic malformation and may explain inconsistent results of surgery in the same category of patients in same hands. The displaced sphincter apparatus was indentified and confirmed to be so by the histolo- gical studies. It was repaired to give high continence rate in cases of incontinent epispadiatics and exstrophic bladder cases in a single stage repair in an unselected series.

Immunohistochemical studies for analysing the presence and development of the neurotransmitters/neuromodulators-substance P (SP), leu-enkephalin (ENK), serotonin (SER), glutamate (GLU), vasoactive intestinal polypeptide (VIP), and cholecystokinin (CCK)-have been conducted on the lateral geniculate nuclear (LGN) complex of albino rats at gestational day 18 (E 18), various postnatal ages and in the adult. The present study shows the unequivocal presence and the sequential changes in the profile of substance P, serotonin, enkephalin and glutamate while VIP and CCK were not demonstrable at any of the age periods. SP immunoreactivity is found to increase from E18 upto 20 day post- natal (DPN) and decrease thereafter to adulthood whereas the SER and ENK immunoreactive fibres and terminals seen as occasional fibres at 1, 5 and 10 DPN are better visualised from 20 DPN and gradually increase upto the adult period. Immunoreactivity of GLU fibres and cells is seen at 40 DPN and increases in the adult. No VIP and CCK immunoreactive fibres or cells are seen at any age period. The vaxing and waning of the undecapeptide SP during the postnatal period of rat besides indicating an evolutionary process may reflect its trophic role in the LGN complex during development. This prompts speculation of a possible therapeutic future in conditions of neuronal degeneration and atrophy encountered in this region in clinical practice. The presence of SP in the adult though in decreasing intensity and SER, ENK and GLU suggests their role in visual processing in dLGN and perhaps in endocrine and behavioural functions of IGL and vLGN. SER, ENK and GLU appear later in this sequential order and increase with maturity. Further studies are required to analyse the role of the various neuropeptides and neurotransmitters existing in the afferent pathways of the LGN complex.

An investigation was undertaken to study the cellular changes following focal ischaemia in Macaca radiata monkeys. The electron microscopic investigation of the structural changes in the caudate nucleus and the insula of the brains from monkeys with right middle cerebral artery (RMCA) occlusion for half, 4, 12, 24 and 48 hours and 2 weeks and one monkey with 12 hours of reflow following 12 hours of occlusion confirmed our earlier observations. Upto 24 hours the edema is only cytogenic and only after that period it becomes vasogenic. Blood reflow following 12 hours of RMCA occlusion does not benefit the tissue. The presence of paracrystal- line array in the vascular endothelium only at half, 4 and 12 hours perhaps gives a clue to the agents which could be the signal in stroke pathophysiology. The sudden change in shear stress on occluding RMCA could have triggered an increase in serotonin which polymerized the actin or an immune reaction which resulted in deposits of immunoglobulins.

The study was carried out to determine the usefulness of air oxygen as carrier gas in balanced anaesthesia technique. A total of 60 patients of both sexes in the age group of 16-60 years with an ASA grading of I/II who were to undergo anaesthesia for more than one hour were selected for the study. Anaesthesia was given from the Pedius B Usha Drager anaesthesia apparatus. The patients were divided in two equal groups; one group on spontaneous ventilation and the other on contro- lled ventilation using pancuronium as per requirement of surgery. Clinical parameters remained within normal limits during anaesthesia in both groups. Recovery in both groups was good and the incidence of post-operative complications was low. The awakening time was 5.12 + 0.57 min in patients breathing spontaneously compared to 2.32 + 0.84 min in those on controlled ventilation. No patient had awareness of any intraoperative event. The use of air as carrier gas may be superior to nitrous oxide as the side effects encountered while using nitrous oxide were not seen. Further, the Pedius B machine can be easily oeprated in remote areas. The disadvantage of using air as carrier gas was in the form of loss of mild analgesic and anaesthetic effects of nitrous oxide necessitating higher doses of volatile and intravenous anaesthetics. The Pedius B machine also had some disadvantages such as water condensation, argon accumulation etc. with the oxygen concentrator.

Urinary excertion of vanillye mandelic acid(VMA) & homovanillic acid (HVA) was estimated in 108 patients with parkinsonism and in 39 controls for VMA & 22 controls for HVA. Urine was collected in acidified containers. The VMA and HVA levels in both morning and 24 hr collection showed positive correlation. There was no significant difference in daily HVA and VMA levels of normal controls. Although there was a decline with age in HVA and VMA levels the difference was not significant. In normal healthy individuals no significant difference in HVA and VMA levels was found between males &femals Urine sample stores at -20 deg.C showed no decline in HVA and VMA levels for several weeks. Amongst 108 patients with parkinsonianism, analysis was done in 43 patients before initiation of therapy and in 65 patients after therapy. In 10 patients follow up was done. The drugs used were Sinemet, Pecitane, Benspar, Desrenyl and Eldepsyl. No significant difference was found between normal control & Parkinson's patients either before or after treatment. The mean excretion of HVA in normal controls, patients on drugs & patients not on drugs was 2.9 + or - 2.6, 11.5+ or - 9.9, 4.3 respectively. Urinary HVA levels were significantly higher in treated patients while there was no difference in untreated patients.

Study was aimed at finding possibilities of preventing or reducing atherosclerosis by the intake of diet rich in fish oil. The fish oil for this study was obtained from the flesh of marine fish, Hilsa ilisa. Atherosclerosis was induced in rabbits experimentally and status of atherosclerosis thus induced was monitored. Investigations were carried out to find out the potentiality of hilsa fish oil rich diets in preventing such induction in rabbits. Future investigations were undertaken on the possibilities of using diets rich in hilsa fish oil in reducing the atherosclerotic conditions in rabbits in which this disease has already been induced experimentally. Studies on the role of lipid components, particularly the fatty acid composition of hilsa fish oil rich diet in preventing and curing atherosclerosis in rabbits were also taken up. The results of the experiments showed that feeding rabbits with hilsa oil supplemented diets, imparted in these animals a resistance towards the deposition of atherosclerotic plaque in the thoracic aorta and coronary arteries, even in the presence of atherogenic concentration of cholesterol and tryglycerides in plasma and platelets. This effect seems to be related to the rise in the proportion of fatty acids of n-3 series of 20 and 22 carbon chain lengths, particularly that of EPA, in the lipids of plasma and platelet due to such feeding. The results of the study showed that feeding atherosclerotic rabbits with diets containing about 10% of hilsa oil, inspite of the presence of 0.45% cholesterol, can regress the atherosclerotic conditions in terms of reducing the atherogenic factors like the contents of plasma total cholesterol, plasma tryglycerides, plasma LDL-cholesterol and platelet cholesterol load on one hand, and elevating the antiatherogenic factors like plasma LDL-cholesterol content and proportion of phospholipid carried to LDL, on the other. This diet can actually regress the thickly deposited fatty material from the thoracic aorta and coronary arteries of the atherosclerotic rabbits. This reversibility of atherosclerotic plaque by dietary treatment is a very important new finding of the present study. The results of the study thus reveal that a hilsa oil rich diet may be useful for the prevention and abatement of hyperlipidemia and atheros clerosis in man and that raw hilsa oil might be effectively used as therapeutic agent in the management of hyperlipidemia and atherosclerosis in human patients.

The beneficial effect of pyridoxine and magnesium oxide administration for eight weeks was studied in hyperoxaluria induced (fed 100mg sodium glycolate/100g bw/day) young adult male wistar rats. Significant hyperoxaluria, hypocitraturia decrease in specific activity of liver transaminase with a concomitant increase in the specific activities of liver glycolic acid oxidase (GAO), glycolic dehydrogenase (GAD) and lactate dehydrogenase (LDH) was observed in sodium glycolate fed group; while pyridoxine supplementation (2.0 mg/100g bw/day) to this group lowered urinary oxalate by 43% and increased citrate by 24%, it also significantly decreased liver GAO and normalised transaminase and GAD. Combined treatment of pyridoxine (2 mg) and MgO (150 mg) per 100g body weight per day to sodium glycolate fed rats normalised hyperoxaluria which is beneficial in retarding calcium oxalate urolithiasis. The clinical study which included assaying the blood and urine specimens of 19 stone-formers after 0, 30 and 60 days of a combined administration of MgO (500 mg/day) and pyridoxine-HCl (10 mg/day) confirmed the beneficial effect of this therapy by lowering hyperoxaluria and promoting citrate excretion, both of which are important factors regulating calcium urolithiasis.

Induction of hyperoxaluria and calcium oxalate calculi was carried out in rats by intraperitoneal injection of hydroxyproline (OH-proline) and feeding hydroxyproline in diet. The effect of Ayurvedic preparations viz. varun and chota gokhura on dissolution of renal calculi, urinary excretion of calcium, oxalate, inorganic phosphorous, uric acid, citrate and inhibi- tory activity towards calcium oxalate monohydrate (COM) crystal growth, enzymes of oxalate biosynthesis viz. glycolic acid oxidase (GAO), glycolic acid dehydrogenase (GAD), lactate dehydrogenase (LDH) in liver and kidney, calcium and oxalate uptake by intestinal BBMV and calcium and oxalate up- take by renal tubular BBMV was studied to assess any alteration in renal handling of calcium and oxalate. Sixteen rats were taken and divided into four groups each containing 4 rats. Twenty four hours urine of all rats was collected at 0 day (Basal) by placing them individually in metabolic cages. 4-OH-L-Proline (2.5g/kg B.W.) was administered intraperitonealy (10ml/kg B.W) to all rats. Group I: animals were sacrificed after 1 day of OH-proline injection; Group II: animals were sacrificed after 2 days of OH-proline injection; Group III: animals were sacrificed after 7 days of OH-proline injection; Group IV: animals were sacrificed after 15 days of OH-proline injection. Twenty four hours urine specimens were collected from individual animals prior to sacrifice at specific time intervals. Liver, Kidney and Intestines were excised at sacrifice and placed on ice. Twenty four hour urines collected were analysed for creatinine, uric acid, phosphorous, calcium and oxalate. Calcium oxalate crystals formed in kidneys after three consecutive intra- peritoneal injections of hydroxyproline (2.5/kg B.W) were examined under polarised light microscope. One rat was sacrificed after each injection and kidneys were removed to examine microscopically, the presence of calcium oxalate crystals. Urine samples of remaining rats were collected after 5, 10, 15 and 20 days of 3rd injection to ascertain the level of hyperoxaluria, which declined between 10 and 20 days. Therefore, to maintain hyperoxaluric level during the drug trial, sodium glycolate (100 mg/100 gm B.W) was orally administered (by gastric intubation) after 10 days of 3rd OH-proline injec- tion. The dried fruits of chota gokhuru were taken and ground and from the co- arse powder an extract was made. To one part of powdered drug, sixteen parts of water was added. The mixture was boiled till 1/4th was left. The contents were filtered through muslin cloth and evaporated to dryness. An extract was made from the residue with Gum Acacia (1-2% in DDW) and tween-20 (1-2 drops per dose of 5g/kg B.W). The drug was administered by gastric intu- bation in morning hours. Eighteen rats were given pellet diet and water ad lib and divided into 3 groups, each of six animals; Group I: were normal saline treated, Group II: were OH-Proline + sodium glycolate + gokhru treated and Group III: were OH-Proline + sodium glycolate treated. Twenty four hour urine of each rat was collected at i) 0-day ii) after 3 injections and iii) after 1 week of 3rd injection and analysed for creatinine, oxalate phosphorous, calcium and uric acid. On the 11th day of 3rd injection, sodium glycolate (100 mg/ 100 gm B.W) was orally administered to Group II and Group III animals. Chota Gokhuru extract was administered to Group II animals after 3 days of sodium glycolate feeding. Before starting the drug, 24 hour urine was collected to check the oxalate level in urine. One intraperitoneal injection of OH-proline to male rats (B.W 200-250 gm) significantly increased in the urinary oxalate excretion within 24 hours of injection. However, the other parameters (viz. calcium, phosphorous and uric acid) were not altered. OH-proline administered for 3 consecutive days showed a significant decline in the oxalate excretion after 10-20 days. Sodium glycolate feeding (100 mg/100 gm B.W) started subsequently after 10 days of three OH-proline injections, was able to maintain hyperoxaluria for next 30 days, to enable to study the effect of the drug. Drug (Tribulus terrestris) administered orally gastric intubation was able to significantly lower hyperoxaluria within seven days and remained unaltered thereafter. Calcium oxalate crystals viewed under polarised microscope after the OH- proline injections were not observed after drug treatment similar to their sodium glycolate fed controls indicating thereby that the dissolution of P calcium glycolate crystals was not due to the drug administration but due to the loss tapering of OH-proline effect. It may be concluded that chota gokhuru administration was not effective for the dissolution of calcium oxalate crystals but helped in lowering hyperoxaluria in experimental model.

Cadmium is known to be potentially toxic to most of the organs of the adult animals. Under normal circumstances very little cadmium reaches the brain in adults because of the selective permeability of blood brain barrier. The susceptibility to cadmium toxicity has been recognised to be influenced by a number of physiological and nutritional factors. One such factor is ethanol whose influence on cadmium neurotoxicity is not widely reported. Study was carried out to find the structural and functional alterations in young adult rat brain after co-exposure to cadmium and alcohol. Cadmium (1mg/kg body weight) and ethanol (2g/kg body weight) were administered alone as well as in combination to different groups of rats, intraperitoneally for a period of one week. Exposure to either cadmium or ethanol alone did not have any significant effect on the body weight but caused a detrimental effect on the growth of the animals when given in combination. Only cadmium exposure caused an appreciable accumulation of cadmium in brain. This accumulation was further enhanced significantly (3- fold) when cadmium was given alongwith ethanol. The increase in cadmium accumulation was maximum in corpus striatum followed by cerebral cortex and other regions of the brain. Marked increase in the uptake and accumulation of cadmium observed after co-exposure to cadmium and ethanol led to a significant depletion in the levels of zinc and copper in these two regions of the brain. Exposure to either cadmium or ethanol alone did not have any significant effect on lipid peroxidation and antioxidant defense system in any of the regions of the adult rat brain but when given in combination, caused a significant increase in lipid peroxidation and markedly decreased the activities of antioxidant defense enzymes like glutathione peroxidase, superoxide dismutase, catalase and glutathione reductase and the levels of glutathione (GSH) in corpus striatum and cerebral cortex regions of the brain. Total thiols and ascorbic acid were reduced only in corpus striatum. Exposure to cadmium alone did not show any significant effect on lipid composition of the brain, but a marked decrease in total lipid (attributed to the reduction in the levels of myelin specific lipids like non- esterified cholesterol, phospholipids and galactolipids) was observed in corpus striatum and cerebral cortex after co-exposure to cadmium and ethanol. In contrast, a significant increase in total lipids, phospholipids, galactolipids and cholesterol was observed in various regions of the brain after exposure to only ethanol. The ratio of cholesterol to phospholipids, a major determinant of membrane fluidity, decreased in these regions of the brain after exposure to only ethanol as well as combined exposure to cadmium and ethanol. Structural alterations observed in terms of increased lipid peroxidation and decreased lipid contents after cadmium and ethanol co-exposure had a profound effect on the activities of membrane bound enzymes such as (Na+-K+)-ATPase and acetylcholinesterase indicating functional impairment. The results of the present study imply that ethanol renders the adult brain more susceptible to cadmium neurotoxicity. Corpus striatum and cerebral cortex are more vulnerable to the toxic effects of cadmium under the influence of ethanol than other regions of the brain.

Study was carried out to delineate the mechanism of action of antituberculous drugs, isoniazid (INH), rifampicin (RIF) through glutathione (GSH) and related enzymes in mycobacteria. The presence of GSH, its metabolic enzymes viz. gamma glutamyl transferase (GGT), gamma glutamyl cysteine synthetase (GGCS) and glutathione synthetase (GSH) and excretion of GSH out of the cells has been established in M. smegmatis. The presence of GSH was established in M. smegmatis both by chemical and enzymatic assays and was further confirmed by covalent binding of N-ethylmaleimide to cellular GSH. Maximum GSH content was observed on second day of the growth which decreased steadily with the age of culture. It was observed that 80 to 90 percent of nonprotein thiols were present in the form of GSH. The activities of four GSH metabolizing enzymes viz. GGT, GGCS, GSH synthetase and GSH-S-transferase were established in 10,000xg and 100,00xg super- natants of M. smegmatis. GGT of M. smegmatis is a peculiar enzyme in demonstrating only hydrolytic activity and not the transpeptidase activity. M. smegmatis cells excrete non protein thiols (NPSH) into the medium which was observed to increase steadily from second to seventh day of growth. Isoniazid (INH) and rifampicin (RIF) deplete cellular GSH in M. smegmatis but have a diverse mode of action. Depletion caused by INH is significantly higher as compared to RIF.NIH dependent depletion of cellular GSH in M. smegmatis is due to an increase in the hydrolysis of GSH as demonstrated by increased activity of GGT, increased activity of glutathione-S-transferase, a decrease in its biosynthesis, as demonstrated by decreased activities of GGCS and GSH synthetase and finally increased export of GSH out of the cell. However, RIF dependent depletion in GSH was due to a decreased biosynthesis of GSH as demon- strated by decreased activities of GGCS and GSH synthetase in M. smegmatis. RIF was found to have no effect on the hydrolysis of GSH and its export out of the cell.

Studies were undertaken to find out the correlation of antifungal activity of imidazole derivatives with cAMP and lipids of dermatophyte i.e. Microsporum gypseum. Cyclic AMP levels were modulated by different modulators and the effect of modulated cAMP levels were studied on lipid metabolism and susceptibility of cells to imidazole derivative. Theophyl- line, prostaglandin E (PGE), dibutyl cAMP, aminophylline, fatty acids and glycerol increased the intracellular levels of cAMP while atropine decreased it. The increase or decrease in cAMP levels was due to the effect of different modulators on the activity of either adenylate cyclase or phosphodiesterase or on both of these enzymes. The lipid composition of M. gypseum was examined in modulated cells and phospholipid content was observed to increase in the presence of theophylline, PGE, dibutyryl cAMP and aminophylline and it decreased with atropine while no change was seen with fatty acids and glycerol. These observations were further corroborated by incorporation of (14 C)-acetate into total phospholipids in the presence of theophylline, dibutyryl cAMP, PGE1, aminophylline and atropine. There was net increase in the synthesis of total lipids and total phospholipids in the presence of above modulators except in case of atropine where decreased synthesis was observed. The increased and decreased activities of key enzymes of phospholipid biosynthesis i.e. glycerol kinase and acyltransferase in case of aminophylline and atropine respectively further confirmed the correlation of phospholipid synthesis and concerned enzymes. The susceptibility of cells to clotrimazole decreased in the presence of various additives (aminophylline, atropine and unsaturated fatty acids). Less susceptibility is due to more resistance of the cells to this antifungal agent in the presence of cAMP modulators. It has been concluded that there is a correlation between intracellular cAMP levels and lipid biosynthesis and lipids play some role in the anti- fungal action of imidazole derivative.

Incubation of human platelets with cholesterol-poor, Cholesterol- normal and Cholesterol-rich liposomes revealed that: acquisition or depletion of platelet membrane-cholesterol was highly selective; variation in membrane cholesterol content with respect to values found in unmodified normal platelets, was paralleled by the observed changes in cyclic nucleotides, amiloride sensitive cytoplasmic pH, phosphorylation of 2l kD protein as well as phospholipase 'A2' activity and no change in phosphoinositol metabolism and the membrane cholesterol modulated changes in cyclic nucleotides, Na+/H+ exchange and phospholipase `A2' activity was completely inhibited when platelets were pretreated with quinacrine (specific phospholipase `A2' inhibitor) before exposure to various liposomes. Based on these results, we suggest that membrane-cholesterol modulated phospholipase `A2' activity may be the basic mechanism responsible for hypersensitivity of platelets in hyper- cholesterolemic patients (Type IIa). Further we also propose that this phospholipase `A2' activity may be regulated by 2l kD protein phosphory- lation/dephosphorylation mechanism initiated by variation in platelet membrane-cholesterol content.

The determination of oxalate in urine and plasma by oxalate oxidase from mosses, barley, banana peel and beet stem requires the pretreatment of urine and plasma to remove cations and anions found in these biological fluids, which complicate the procedure and consequently the sensitivity and reproducibility of the method suffers. An oxalate oxidase has been purified from leaves of 10-day old seedling plants of grain sorghum CHS-5, which is insensitive to these urinary inorganic ions viz. Na+, K+, Ca++, Mg++, Fe++, Zn++, Pb++, Cl-, PO4--, SO4--, HCO3-, CO3-- etc. These improved characteristics of the sorghum enzyme make it ideally suitable for its use in the deermination of oxalate in urine and plasma. A simple enzymic colorimetric method for determination of urinary and plasma oxalate has been developed using the sorghum enzyme. The method simplifies the total analytic procedure by eliminating the need for a separate step to remove cations and anions from urine plasma, prior to oxalate assay. The method is rapid and does not require expensive specialized equipments and therefore suitable for oxalate anasysis in a large number of samples in clinical laboratories.

Investigation were carried out to study the correlation of drug induced superoxide radical production, lipid peroxidation methemoglobinemia as well as interaction of superoxide dismutase and ascorbic acid. Ascorbic acid is a potential scavenger of superoxide radical. Some commonly used drugs like sedatives, narcotics, antiamoebics produce increased superoxide in the liver microsomes of rats and guinea pigs by the NADPH - cytochrome P450 reductase - cytochrome P450 electron transport system, commonly known as the mixed function oxidase. Rats can synthesize ascorbic acid. These drugs stimulate ascorbic acid production in the rat apparently to scavenge the increased amount of superoxide formed. In the guinea pig, a species incapable of producing ascorbic acid, administration of these drugs results in superoxide toxicity, as evidenced by lipid peroxidation in diferent tissues including liver, kidney, lung, heart and adrenal glands. Lipid peroxidation is highly detrimental to structure and function of the membranes. Ascorbic acid completely prevents lipid peroxidation both in vitro and in vivo. Preparation of microsomes, estimation of superoxide radical, assay of cytochrome reductase, cytochrome P-450 content, estimation of ascorbic acid and lipid peroxidation were carried out using standard procedures.

Study was undertaken with the aim to identify the molecules and molecular mechanism(s) involved in the development of human atherosclero- sis and to study the characteristics of foam cells and their transport into the sub-endothelium. For this, procedure for harvesting and culture of human umbilical arterial endothelial cells was optimised. Scanning electron microscopy revealed homogeneous population of endothelial cells in culture. On interaction with endothelial cells substantial structural changes were noticed in human plasma low density lipoprotein (LDL). Changes in biophysical environment of several amino acids of modified LDL were also noticed by circular dichroism, fluorescence and electronic absorption spectrosopic methods. Lipid peroxidation was measured by second derivative electronic absorption spectroscopy and higher levels of conjugated dienes in LDL and extractable lipids of endothelial cells were found. Circular dichroism, derivative fluorescence and electronic absorption spectroscopic methods established that the circulatory monocytes/macrophages utilised endothelial modified as well as chemically modified LDL and degraded it. Scanning electron microscopic and fluorescence microscopic observation suggested that upon interaction with modified LDL, macrophages got transformed into lipid laden cells (foam cells), about 4-times larger than the macrophages. Presence of foam cell like structures was noticed in atherosclerotic plaques. Results of the present study suggest that at the molecular level the partially modified apo B and peroxidised lipids of LDL are involved in the pathogenesis of human atherosclerosis.

Forskolin, a diterpenoid, reported to stimulate melanocyte prolifera- tion in vitro, was tested to find out its effectiveness in vivo. For this purpose an experimental model was designed in which the melanocytes in the skin (of guinea pig) are selectively destroyed by local application of 4-methoxyphenol, thereby resulting in a gradual loss of pigment in the skin, the condition being similar to vitiligo in human beings in several respects. Following cessation of application with 4-methoxyphenol, the remaining peri-lesional and follicular melanocyes proliferate and reoccupy the dermal-epidermal junction in the depigmented patch and this is reflected visually by the appearance of pigment. When forskolin was topically applied on the depigmented patch at a dose of 7.5 X 10 moles, 3 times a day in glycerine base, no enhancement was found in the rate of repigmen- tation compared to only base applied controls. However, following intradermal injection of 4X10 moles of forskolin, 2 times a day, into the depigmented patch, it was found that the per cent of completely pigmented ear lobes after 12 weeks of treatment was 75% of forskolin treated ear lobes against only 43% in controls. Forskolin was thus found to exert its stimulatory influence on melanocyte proliferation under in vivo conditions also.

Trophoblast and the tissues of th uterus, together from an important organ, the placenta, which provides the growing embryo with nutrition and with oxygen; it also removes waste products from the embryo. It is proposed to study the regional distribution of zinc, copper and cadmium in human fetal membranes and placenta. Isolation and purification and structural characterization of metal binding protein(s) from human placenta and fetal membranes will be done. Antibodies will be raised agaisnt the major metal binding protein(s) and homology of this protein with other metal binding proteins in liver and kidney will be determined besides studies on the role of metal binding protein(s) in detoxification mechanism.

Liposomes have been used to deliver a wide variety of biologically active substances to different tissues in vivo. Attempts have been made to deliver liposomes to brain with very limited success beacause of the obstacle imposed by the blood-brain barrier. In this study, attempts were made in the delivery of substances entrapped in multilamellar and uni- lamellar liposomes (phosphatidylcholine: cholesterol: sulfatides, 7:2:1) using 2M mannitol to disrupt the blood-brain barrier.Six to 14-fold increase in the uptake of CAMP phosphodiesterase, a high molecular weight protein, and a 3-fold increase in the uptake of CAMP, a low molecular weight compound was observed in the presence of mannitol. Delivery of entrapped materials to the brain was confirmed by immunofluorescence studies using morphine-BSA. 2M mannitol also resulted in a slow clearance of both free or liposomally- entrapped L25I-PDE fraom brain. In vitro studies using C6 glioma cells showed increased incorporation of 32P label from (Y-32P) ATP into TCA insoluble pellet in the case of liposomal CAMP-protein kinase-(Y-32P) ATP complex as compared to free complex at all time points studied. Studies with metabolic inhibitors along with small unilamellar vesicles (SUVs) containing either morphine-BSA or FITC-BSA indicate that one of the mechanisms of uptake of liposomes by these cells could be endocytosis although fusion also appears to take plac

In all 561 patients affected with peptic ulcer (511 were suffering from DU and 50 with GU) were selected for this study in the duration of 3 years. The endoscopic findings were recorded in a proforma along with other details. All the endoscopically proven peptic ulcer cases were selected for the present study. Information regarding age, sex, dietary habits, smoking, alcoholism, pan chewing and incidence of ulcer were obtained. From all these patients blood and urine samples were collected for biochemical studies. The proportion of affected individuals in all the pedigrees was determined based on the segregation of the condition among the sibs of the propositil where (a) both the parents of the propositil were affected (b) one of the parents was affected and (c) both the parents were normal. Out of 365 cases, 70 (19.18%) were found to be vegetarian and the remaining 295 (80.82%) were non-vegetarians. The percentage of non-vegetarians was foun to be higher. Out of the 544 cases 212 (38.97%) were found to be regular in taking meals when as 332(61.03) were irrugular. The familial and isolated groups of patients differed significantly with regards to the frequencies of cases laking diet regularly and irregularly. the percentage of panchewing cases in 4 groups of DU patients i.e. male femilial, female, familial, male isolated and female isolated were 52.54, 23.73, 18.64 and 5.08 respectively and with tobacco 67.24, 22.41, 8.62 and 1.58. the precentage of excess consumption of chillies and spices in food in 4 groups as mentioned above is 21.83, 2.11, 52.82 and 23.24 respectively. Moderate consumptions of chillies and spices were found to be 16.95% 3.39%, 58.47% and 21.11% respectively in the above mentioned groups. The data on cigarette smoking was available in 516 cases. 300 (58.14) were found to be smokers and 216 (41.86) were non-smokers. A higher precentage of smokers was found to be in isolated male patients. With regards to alcohol consumption out of total 437, 111 (25.40) were alcoholics and the remaining 326 (74.60%) were non-alcoholics. A higher precentage of alcoholics were found in male alcoholics and non-alcoholics. All the pedigrees indicating positive family history of ulcers were examined for identifying mode inheritance of the conditions. To understand the segregation of condition among the family members, parents of the propositii were grouped into 3 categories based on the presence or absence of ulcer in them like 1) AXA (affected x affected) both parents were affected, (2) X N (affected X Normal) one of the parents was affected and (3) N X N (Normal and Normal) both the parents were normal. The proportion of affected sibs was higher 50.00% in the sibships where both the parents were affected as compared to those, where one parent was affected (30.85%) and none of the parents were affected (29.31%) The mean unit pepsinogen activity per ml. per 24 hours values of serum and urine for DU patients, and GU patients and then first degree relatives are given in tables 15 and 16. The mean serum pepsinogen concentration in patients (male & female) familial and nonfamilial were significantly elevated where compared with that of controls. (p>0.001). A similar observation was recorded in uropepsinogen values of DU patients. Where as Uro and serum pepsinogen levels in GU was significantly elevated. Out of 310 serum pepsinogen values 188 (60.64%) were found to be normopepsino- genemic. DU patients and the remaining 122 (39.35%) were hyperpepsinogenemic . Out of 310 uropepsinogen values 48 (15.48%) found to be normal and 262 (84.52%) were with hyper uro pepsinogenuria. uropepsinogen was estimated in general population (720 subjects) of Hyd erabad. Out of 335 symptomatic subjects 130 (38.80%) wer found to be with hyper pepsinogen values and the remaining 191 (57.01) were normal values. Out of 385 asymptomatic subjects 72 (18.70%) were found to be hyper pepsinogenemic and remaining 327 (84.93) were within normal range the relative risk of develop ulcer was found to be 3.1 in individuals with elevated uropepsinogen levels.

Phosphorylation and sulfation of proteins are two means of post- translational modification of proteins. While phosphorylation takes place on tyrosine, serine or threonine residues, sulfation takes place on tyrosine residues alone on the proteins. Phosphorylation serves many important functions such as activation or inactivation of enzymes and cellu- lar signalling receptors. Sulfation of proteins is known to alter the biological activity of proteins and promote the fast axonal transport of proteins. In the present study, phosphorylation of a non-glycoprotein enzyme, a cytoplasmic neutral alpha-D-mannosidase from monkey brain has been demons- trated. Phosphorylation on serine residues is catalyzed by a cyclic AMP dependent protein kinase. Phosphorylation also alters the cobalt ion sensitivity of the enzyme. An aminopeptidase from monkey brain purified by investigators earlier has been found to cleave enkephalin sequentially to its constituent amino acids. This aminopeptidase is phosphorylated on serine residues by cyclic AMP dependent protein kinase resulting in its inactivation. It has been demonstrated by means of chemical modification of amino acids that tyrosine residues are involved in the catalytic activity of the enzyme. Sulfation of proteins has been demonstrated in the monkey cerebellum and young (10 days old) rat brain. The sulfotransferase respon- sible for transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate was isolated from monkey cerebellum. Both endogenous proteins and exogenous poly(Glu,Ala,Tyr) were sulfated. Using lectin-sepharose affinity chromatography, it was found that both mannose and galactose containing glycoproteins could undergo sulfation. Evidence for differences in the polysaccharide chain structures of the sulfated glycoproteins from monkey cerebellum and young rat brain was also obtained. Modification of tyrosine residues in proteins resulted in partial abolition of phosphorylation and full abolition of sulfation.

Among the fungal toxins, 'aflatoxins', produced by Aspergillus flavus and A. parasiticus group of fungi have received worldwide attention, due to their potentialities as carcinogenic agents in various laboratory and far animals. Various analytical methods have been developed for the detection of aflatoxins in foods, feeds and biological fluids. These include physico- chemical and immuno-analytical methods. The primary objective of the study was to develop a simple homogeneous enzyme immunoassay for the detection of aflatoxin B1 in biologi- cal samples. In the first phase aflatoxin specific antibodies were produced in rabbits. Initially, the requisite antigen i.e. BSA-AFB1 and Poly-D-lysine-AFB1 conjugates were synthesized for raising antibodies. These antibodies were used for aflatoxin detection, either by homogeneous immunoassay or by heterogeneous immunoassay (ELISA). The enzyme horse radish peroxidase (HRPO) was used as a probe for homogeneous immunoassay. The conjugation of aflatoxin B1 to HRPO resulted in 54-72% loss in enzyme activity. In the presence of aflatoxin specific antibodies, the HRPO-AFB1 conjugate showed reversal of its lost enzyme activity (12%). This was used as an analytical tool to quantitate aflatoxin B1. The displacement studies carried out with free aflatoxin B1 and HRPO conjugate in the presence of antibodies indicated poor linearity as compared to the heterogeneous assay. The number of lysine residues available for conjugation in HRPO were determined to be g, based on a novel method developed. However, it was observed that 6-8 residues were involved in conjugation with AFB1. The low level of modulation of enzyme activity by antibody, with respect to HRPO-AFB1 conjugate, could possibly be explained by the limited number of lysine residues on HRPO molecule and its proximity to the active site of the enzyme. Thus, HRPO was found to be limiting as an enzyme label with respect to homogeneous enzyme immunoassay system for aflatoxin B1 analysis.

Investigations were undertaken to study the effect of essential fatty acids (EFAs) and their metabolites on free radical generation and lipid peroxidation to investigate their possible effect on normal and tumor cell membranes by studying their effect on cell membrane bound enzymes such as Na+K+-ATPase and 5'-nucleotidase and on DNA and DNA synthesis in normal and tumor cells in vitro. Normal monkey kidney (CV-1), normal human fibroblasts (CCD-41-SK), human breast cancer (ZR-75-1) and human cervical carcinoma (HeLa) cells were cultured. To study the growth of various normal and tumor cells in the presence of different concentrations of various types of fatty acids, thymidine incorporation studies were undertaken. Experiments were performed with cyclo-oxygenase inhibitor, indomethacin, lipoxygenase inhibitor, NDGA; anti-oxidants, BHT, vitamin E, SOD, different types of prostagladins and calmodulin antagonists to study the mechanism of the tumoricidal action of GLA and other fatty acids in vitro. Assay of superoxideanion and other free radicals was done by nitroblue tetrazolium (NBT) reduction method. The total amount of lipid peroxidation products present in the cells was estimated using thiobarbituric acid (TBA) method. To know whether cis-unsaturated fatty aicds (c-UFAs) can injure cell membrane and thus, bring about their cytotoxic action, the effect of various fatty acids on the activity of membrane bound enzymes such as Na+-K+-ATPase and 5' nucleotidase of tumor (HeLa) cells was studied. The result obtained from the studies indicate that EFAs and their metabolites especially DHA, EPA and GLA can kill tumor cells selectively. They augment free radical generation and lipid peroxidation in ZR-75-1 and HeLa Cells. The cytotoxic action of EPA, DHA and GLA on HeLa cells can be blocked by anti-oxidants such as vitamin E, SOD, BHA and BHT and also by cyclo-oxygenase and lipoxygenase inhibitors namely indomethacin and NDGA as well as by calmodulin antagonists. Inhibitors of the cytotoxic action of GLA and other c-UFAs can also inhibit the production of free radicals and lipid peroxidation process in HeLa cells induced by these fatty acids. Hydroxy and hydroperoxy products of lipoxygenase system derived from GLA but not prostaglandins showed cytotoxic action against HeLa cells. On the other hand, prostaglandins enhanced HeLa cell growth. These results suggest that lipoxygenase products are the mediators of the cytotoxic action of GLA on HeLa cells. Studies performed have thus demonstrated that GLA and other c-UFAs have selective cytotoxicity against tumor cells but do not harm the normal cells. These fatty acids are able to bring about their selective tumoricidal action by enhancing the generation of free radicals and lipid peroxidation process in the tumor cells.

Studies on the neurobiochemical mechonisms in development of behavioural tolerance to methylxanthine and its withdrawl effect Study was carried out male charles Foster strain albino rats to understand the neurobiochemical mechanism explaining the development of behavioural toleronce to methylxanthine (caffine) and its withdrawl effects at the level of (i) the dynamics of different neurotransmitter viz, GABA and adenosine receptor binding and (ii) introduction of GABA, adenosine with receptor of other neurotransmitters. Rats were divided into 3 groups and animals of group 1 and 2 were treated with caffeine at a dose of 10 and 20 mg/kg/day respectively for 1-16 consecutive days. Group 3 animals (control) were treated with water (vehicle) only Single does (10-20mg/kg) of methylxanthine (cafeeine) increase the locomotor acitivity (LA) of rats in a dose dependent manner. The caffeine induced LA incease further with the duration of treament showing a peak after four consecutive treatment. This increased LA was restored back to control values after 16 and 12 consecutive days of caffeine treatment at a dose of 10 and 20 mg/kg/day, respectively indicating the development to tolerance to caffeine. Pharamacological and biochemical studies suggest that caffeine under nontolerant conditions reduces adenosinergic activity by its antagoistic effect on adenosine receptor which in turn enhances dopaminergic activity thereby reduceing central GABA ergic activity through the inhibition of cholinergic system and results in enhanced locomoter acitivity. The development of tolerence to caffine following 12 or 16 consecutive days of treatment depending on the dose was found to be due to up gulatioin of central adenosine receptor activity and GABAergic activity through the modulation of dopaninargic and cholinergic systems. The withdrawl of caffine following its tolerance decreases LA maximally at 48 h from the time of withdrawl. This decrease in LA under cafeine withdrawl was found to be the effect of increase in central GABAergic activity via the modulation of central dopaninergic and cholinergic activities. It is thus concluded that caffeine induced changes in LA under nontolerant, tolerant and withdrawl conditions are mediated by modulation of not only the central adenosinergic acitvity but also the central GABAergic acitvity through the interaction of dopaminergic and cholinergic systems.

The present study was aimed to; investigate the nature as well as degree of oxidative stress imposed by managnese, endosulfan and acrylamide on the nerve cells/tissue; Understand the mechanism of the neuronal tissue injury and understand the role of antioxidative defence capacity of the tissue particularly GSH) in modulating the neurotoxicity of these chemicals. Impairment of oxidant antioxidant balance has been implicated in the etiology of cerebro vascular disorders and upon exposure to a variety of enviromental chemicals including metals, solvents, pesticides and monomers. Considerable progress has been made in elucidating the mechanisms of free radicals induced membrane damage in various tissues and organs, however free radicals induced injury to the brain, upon exposure to environmental chemicals and in ischemic stroke has not been studied extensively. Since reavtive oxygen species and fatty acid hydroperoxides are physiologically present in brain and platelets and believed to play a regulatory role in membrane functions, it was of interest to investigate the possible involvement of reactive oxygen species in altering the membrane related events upon exposure to a metal managanese, monomer acrylamide and pesticide endosulfan and in patients of ischemic stroke. The observations revealed that Iron Ascorbate treatment in vitro, resulted in a significant increase in LPO and depletion in GSH level both in brain and platelets. This was in parallel to decreased membrane fluidity and elevation of (Ca++) i levels. Free radical scavengers mannitol, thiourea and physio Ehysiological antioxidant vit- E provided partial protection in Iron-Ascorbate induced lipid preoxidation and gluthione oxidation and consequent alterations in membtrane fluidity, (Ca++)i levels and platelet aggregation. Acrylamide at concentrations > 5 mM (in vitro) caused a decrease in GSH levels and increase in LPO and membrane function alterations. However, at lower concentrations (<5 mM) at which acrylamide has previously heen ahown to cause neuronal tissue injury neither caused oxidative damage nor altered neuronal membrane function. Acrylamide and endosulfan exposure in vivo although caused a slight increase in LPO, the intgrity and functions of neuronal membranes remained unaltered both in young and adult rats. The observed increase in 3H spiroperidol binding following acrylamide exposures appears to be independent of oxidative events. The significant decrease in LPO and increased membrane fluidity in human platelets and rat brain preparations in vitro and in vivo suggest the antioxidant like action of manganese. The increased levels of PC and PE, increased 3H spiroperidol binding and deoreased 3H- 5HT binding in young rats as compared to the adults suggest that young rats are more susceptible to manganese mediated neuronal membrane damage. Incrased platelet aggregatino in response to ADP, epinephrine arachidonic acid was observed in ischemic stroke subjects as compared to normals. This was in parallel with incrase in platelet (Ca++)i levels, enhanced rate of MDA formation, oxidation of reduced glutathione and decrease in membrane fluidity. From the present observa observations it may be concluded that oxygen free radicals play a significant role in modulating the membrane related events following exposure to environmental chemicals and in pathological conditions such as ischemic stroke.

The study was carried out to evaluate the role of a membrane glycoprotein on the control mechanism of anorexia in normal and diseased condition in swiss albine nice. Bioassay studies revealed the presence of anerexigenic glycoprotein (proteoglycan) in both monocatylednors wheat (Triticum asestivum), rice (Oryza sativa) and barley (Hordeum valgare) as well as dicotyledorions mung sean (Phaseolus radiants and chick pea (Gicer orietinum) plants. However, the proteogly can isolated from the plants of the graminease family (monocot) suggested faster degradation in animal body. A similar loss of appetite response was observed following the administration of protoglycan to the animal model system either by intramuscular/ intravenous/ intradermal/ or intraperitoneal rout. But no anorexigenic response was observed when isolated proteoglycan was administered orally. Majority of the non ionic detergents were not found suitable for the suitable and effective isolation from ming bean sprout preparation. Subcellular fractionation studies clearly revealed its presence in cell surface (plasma) membranes which strengthed the hypothesis that the anorexigenic proteoglycan is anchored to the surface membrane. Overall studies on this anorexigenic proteoglycan suggest that, being a common constituent of the surface membrane it is well accepted by the animal body without any rebound or apparent unwanted effect. Hence it can be hypothesised that with a regulatory control on hunger drive, it could provide a useful tool in the hands of physicians dealing with the problem of obesity (due to overeating) and anorexia in various diseased conditions. In addition, it can also be used in various nutritional research studies as a conventional parallel control group with controlled food intake depicting a true satiety condition.

The study was carried out to elucidate structure and function and biochemical characterization of lectins from human and rat foetal brains. Human factness were obtained from nutritionaly normal ladies undergoing medical termination of pregnancy. Lactose binding (18KDA and 66KDA) as well as beparin binding (29KDA and 45KDA) lectins were pruified from human foetal brain by a simple chromatographic procedure using Q-Sepharose as the matrix. Through both 29KDA and 45KDA lectin were heparin inhihitory, the 29KDA heparin binding lectin showed inhihition towards stachyose as well as mesoglycan while 45KDA hepar in binding lectin showed strong inhihition by sulodexide as well as mesogtlycan. This revealed the complexity of endogenous ligands for 29 and 45KDA lectin. Both 29KDA and 45KDA lectins showed high content of acidic aminn acid. Absence of pysine from 29KDA lectins distinguishes it from other known lectins. The major glycopeptide recognized by 29 and 45KDA lectins was 0.5M N aretyl D glucosamine eluted glycopeptide. Mannose terminated glycopeptide had no effect on these lecitns. Higher contents of 29KDA lectin in cerehral cortex and powest in spinal cord cnrrelates lectin levels with neuronal population. Presence of 29KDA lectin in mitochondrial fraction can be correlated with the lysosomal functioins i.e. as a signal for secretion and release. Myelin basic protein isolated from rat brain showed heparin inlibitery lectin activity. This property can be used to identify the abnormalities of myelin where myelin basic protein leaks in cerebrospinal fluid.

Chemical analysis of urinary calculi in Haryana. The study was carried out to elucidate the morphological characteristics and chemical composition of urinary calculi formed by people of Haryana and to correlate the chemical composition with the type of calculi and age of stone formers. Quantitative chemical analysis of 400 urinary calculi (240 renal, 66 ureter, 94 bladder) collected from different districts of Haryana revealed the presence of calcium oxalate monohydrate (CaOxM) in all the stones. However the presence of magnesium ammonium phosphate hexahydtate (MAP), hydroxy apatite ( HA) and uric acid (UA) was observed in 83.6%, 93.46% and 77.2% patients respectively. The content of CaOxM was found to be higher in renal stones compared to bladder to renal and bladder stones. The concentration of uric acid was found to be higher in bladder stones as compared to renal and ureter stones. The CaOxM was found as a major component in all types of urinary stones analysed. The presence of CaOxM and HA decreased and MAP and UA increased with the age of the stone formers.

The study was carried out on Wistar strain albino rats to investigate the biochemical mechanism of aluminium phosphide (ALD) poisonig with special emphasis on mitochandrial dysfunctions in brain and to find out if Vitamine E has any protective effect on aluminium phosphide induced neurotoxicity. Acute aluminium phosphide (10 mg/kg body wt.) exposure resulted in disruption of energy metabolism by altering the various components of electron transport chain in mitochondria and disturbing the carbohydrate homeostasis. A markeddecrease in the activity of cytochrome oxidase was observed which resulted in the decrease in the oxygen uptake. The ATP hydrolysis which lead to depletioin of ATP levels in mitochondria. The components of the electron transport chain, ie the NADH dehydrogenase and dehydrogenase showed a significant decrease in their activities. This energy crisis resulted in a considerable decrease was obsered in the acitivity of hexokinase in various regions of rat brain. A decrease was also observed in the gluconeogenic enzymes viz Glucose-6-phosphatase and fructose 1,6-bisphosphatase. Glucose-6-phosphate dehydrogenase also depicted a marked decrease. ALP exposure resulted in oxidative stress to the different regions of centralnervous System through lipid peroxidation and antioxidant defense system thereby affecting the structure and functiins of synaptic plasma membranes. There was significant increase in the levels of lipid peroxidation in cerebral cortex, cerebellum and brain stem on exposure to ALP. Superoxide dismutase and catalase demonstrated a marked decrease in all the three regions of rat brain. The acitvity of glutathione reductase was significantly decreased following ALP exposure however, glutathione peroxidase remained unaffected. The levels of reduced and oxidized glutathione decreased significantly and total sulfhydryl groups showed a marked decline. The membrane fluidity was adversely effected due to oxidative stress generated following toxic exposure ALP wshich further affected the acitvities of membrane bound enzymes i.e. acetycholine esterase and sodium and potasium ATPase. In a short term study of 15 days, vitamin E (150 IU/kg body wt./day) was found to decrease ALP(1mg/kg body wt/day) induced oxidative stress in rat brain. Vitamine E and ALP administered together resulted in reduced levels of lipid peroxidation in all the region of rat brain compared to only ALP treated group. The activity of superoxide dismutase was enhanced significantly when vitamine E was given with E. This also resulted in decreased glutathione reductase levels as compared to only ALP treated group. ALP exposure exerts a devastating effect on the energy metabolism and generates severe oxidative stress leading to various structural and functional alterations in the cell. Vitamin E showed some protective role in elevating toxic effects of ALP.

The objective of study uptake and distribution of essential fatty acids and their metabolites in normal and tumour cells in vitro using radiolabelled compounds and the effect of essential fatty acids and their metabloities on the concerntations of various anti-oxidants such as superoxide dismutase, catalase and glutathione in tumor cells in vitro. To characterize various lipid peroxides formed in the tumor cells treated with essential fatty acids and their metabolities by HPLC/GC analysis using dervatization methods. The effect of various fatty acids was seen on the growth and survival of tumor cells in vitro (HcLa cell). The percentage of live cells was less as the dose of fatty acids was increased EPA and DHA were found to be the most potent fatty acids, n-6 and n-3 fatty acids were found to have cytotoxic/ cytostatic action on HcLa cells. ALA & EPA fatty acids were mostpotent in inhibiting the growth of SP2/0 cells at all concentrations tested. DHA and OA were also found to affect the proliferation of SP2/0 cells but only to a limited extent. The n-6 fatty acids: LA, GLA, DGLA and AA did not show any inhibitory effect on the proliferation of SP2/) cell at all doses tested. Infact, both DGKLA and AA Stimulated the proliferation of SP 2/0 cells. Both ALA and EPA also decreased the viability of SP 2/0 cells. Both ALA and EPA also decreased the viability of SP 2/0 cells bout only to a limited extent and that too obly at 40 mue g/ ml concentratioin. KB-Ch R-8-5 cells are resistant to the cytotoxic action of vincristine compared to the response of KB-3-1 cells. Of all the fatty acids, EPA and DHA are the most potent in reducing the survival of KB-3-1 and KB-Ch R-8-5 cells in vitro. The cytostaitc/ cytotoxic action of various fatty acids on KB-3-1 and KB-ChR-8-5 cells in further evident from the thymodine incorporation studies. Studies have demonstrated that the sensitivity or resistance of tumor cells to the cytotoxic action of anti cancer drugs and cytokines is determined, at least in part, by the intracellular anti-oxidant concentrations.Thus one of the factors that can contribute to tumor cell drug resistance is their anti oxidant content was studied. This study was performed in SP2/) cells. Of all the fatty acids tested, only ALA and EPA exhibited potent cytotoxic action on SP 2/0 cells. Hence, the effect of ALA and EPA only on the concentrations of a5t ous anti oxidants in Sp 2/0 cells was studied. Results of this study revealed that ALA and EPA ( 5 and 10 mue/ ml) treatment can decrease the concentrations of SOD(superoxide dismutase) to a significant degree in SP 2/0 cells. ALA treatment decreased where as EPA enhanced the activity of catalase at the end of 24 hours of incubation. A Decrease in the levels of glutathione peroxidase (GPX) and gluthathione reductase (GR) with ALA (5 and 10 mue/ml) treeatment where as an increase with EPA treatment (10 mue g/ mg) was observed. A significant change in the levels of glutathione-S transferase (GST) was induced by ALA (at mue / ml dose. Total glutathione levels were elevated in the cells treated with 10 mue g/ ml of ALA and EPA at the end of 24 hours of incubation. These results indicate th that a marked degree of alteratin in the levels of various anti oxidants in SP 2/0 cells can be induced by ALA and EPA treatment.

Aluminium for a long time has been considered an indifferent element form a toxicological point of view. In the recent years the knowledge about the ubiquatous nature of this element and the growing evidence about its neurotoxicity makes it the subject of increasing interest. Neurological dysfunctions in patients of dialysis encephalopathy have been related to high aluminium concentration in the dialysate and to the use of phosphat binding gels containing aluminium. An attempt has been made to drawn a correlation between the biochemical abnormalities, behavioural aberration and histopathological alterations following aluminium exposure. Male abino rats of Wistar strain in the weight range of 140-160 gms were used throught the study of 12 weeks. The various biochemical parameters to be stud 12 weeks. The various biochemical parameters to be studied were standardized. After the respective treatments, animals were fasted overnight and euthanised by decapitation. The brains were removed, rinsed in ice cold isotonic saline and dissected into the following regions according to the guidelines of glowinski and Iverson (1996), cerebral cortex, corpus striatum and hippocampus. The following experiments were conducted Momory Function Test, Intrasynaptosomal Calcium assay, Calcium uptake in synaptosomes, Calcium ATPase assay and preparation of synaptic plasma membranes from rat brain was also done. Memory function test i.e. Passive avoidance and Active avoidance. The animals following chronic aluminium exposure showed a significant decrease in retention time during the passive avoidance memory test.

The role of placental steorids, hCG, inhibit and opioid peptides in the regulation and secretion of GnRH by placenta was investigated using both human and rat placenta as model. In the first Phase of the study experiments were designed to evaluate the effects of placental steroids in the GnRH levels in the in vitro incubation studies. These studies involved standardization of RIA for GnRH and application of GnRH RIA for monitor the levels of GnRH under in vitro conclusions following addition of modulators. In the second phase of the study experiment were planned to monitor the level of GnRHR mRNA since it is known that at the pituitary level the regulation is mainly mediated by modulating the mRNA level of GnRH recptor by the gonadal steriods and accordingly human placenta. Both in vitro and in vivo studies, which were carried out to evaluate the efficacy of the GnRH receptor antiserum to inhibit binding of GnRH to pituitary, suggested that the antibody is not able to inhibit binding. Hence, active and passive immunization studies were undertaken to evaluate the effects of long term exposure to antibodies under in vivo conditions. The goal of the studies described herein was to use the primers based on the sequence of the human pitutiary GnRH receptor to amplify, clone and sequence transcript of interest and to determine their homology to the hpit GnRH receptor sequence. The data generated would contribute to the understanding of the similatities and differences between the hypothalamic/ pituitary and placental GnRH axes and this inturn would perhaps contribute to our overall understanding of hCG GnRH regulation.

A study on the role of different neurotransmitters in regulating the GI motility, indicates cholinergic involvement and partial role of prostaglandin and NO in the mechanism of action of black tea extract. Results indicate that thearubigin fractin of responsible for the prokinetic effect of black tea extract. Exepanol-HCl (drug under development as a prokinetic agent) produced protentiation of GIT both in normal and in hyperglycemic rat.Exepanol HCl (10 mue gram/kg) reduced significantly indomethacin treated ulcerogenic rat. (upto 82%). Exepanol was equipotent as MCP by restoring the gastric emptying time in normal as well as in hyperglycemic rat. Results indicate a role of NO in the mechanism of prokinetic effect of Expanol HCl. Now to extrapolate the data already generated in our laboratory as mentioned above, we are in the process of estimating NO in different parts of GI tract in control group as well as in experimental group treated with graded dose of prokinetic drug, Exepanol HCl. Studies ith ciprofloxacin (a, broad spectrum fluoroquinnolone antibacterial agent) also indicates an involvment of cholinergic mechanism in the prokinetic effect of ciprofloxacin both in normal and in diabetic rat. It is suggested that systemic clinical studies may be initiated to evaluate the potential efficacy of ciprofloxacin in the treatment of GI motility disorders, specially in diabetic gastroporesis. During the course of investigation, some medicinal plants are evaluated for possible prokinetic and or antidiarrhoeal activity. The results so far indicates that edible mushroom s shares prokinetic effect. Seed extract of albizzia ledbeck and root extract of psidium guajava exhibited marked anti diarrhoeal activity in animal model. Its potency is comparable with loperamide, the clinically used antidiarrhoeal drug. Results indicate that the antidiarrhoeal action of the extract may be due to the inhibition of prostaglandin synthesis and release.

The study was planned and carried out in the Department of Physiology and subjects recruited in OPD of Obst. & Gynae, in St. John's Medical College Hospital. In all 155 patients were included in the study, after obtaining the informed consent as required by the Ethical Committee of the Institution. The subject groups were as follows Control (55) first trimester (n=13) second trimester (n=42, 3rd trimester (n=45) of which normotensive (n=32), PET (n=13). The nitrate levels were measured by one step enzymatic assay using nitrate reducfase method. Fasting blood sugar, Hemoglobin level, creatinine were measured using standard laboratory producers. The value of serum nitrate in all the gropus had a skewed distribution and therefore these werre analysed by non parametric tests i.e. Kruskal Wallis and Mannwhitney by using statistics package SPSS 7.5. The serum nitrate levels were highest during 111o pregnancy (median 22.8, IQR 26.2) as compared to first trimester (median 16.1, IQR 22.4). This is an expected finding. The level of nitrate in Preclamptic group were significantly lower (p=02). The renal function of all the subjects were within normal range, and therefore differences could be attributed to the reduced production of the nitrates from Nitric oxide. Although the number of subjects in the hypertensive group is small but the results of present study indicate lower levels of nitrate production in Preclamptic group. Larger study is needed to confirm and extend the findings.

The present study characterised alpha 1 and 2 adrenoreceptors by using specific radioligands in regions of rat brain. The monophasic displacement by unique pharmacological agents of each subtypes provides strong evidence that atleast two distinct populations exist in rat brain. The denisity of alpha 1 and 2 adrenorecptors was studied in different regions of rat brain after chronic exposure (40 days) to antidepressants (ADs). Tricyclin antidepressants act by downregulating cortical adrenoreceptors but not hippocampal adrenoreceptors. However there was no significant change in the affinity of these receptors. The differential alteration of adrenoreceptors linked second messengers like cAMP and IP3 was also observed in reponse to chronic administration of ADs. The phenomenon of increased synaptic NE content due to blockade of uptake sites by TCAs with concomitant stimlation of post synaptic receptors may be resulted in the subsensitivity of cortical adrenergic signalling. This decreases in alpha adrenergic responsiveness could be the mechanism of action of TCAs and the adaptive phenomenon of desensitisation of receptors may be considered an important factor in the achievement of theraupeutic efficacy of AD drugs after chronic tyreatment. This study has provided a complete profile of biochemically and pharamcologically characterised alpha1 and 2 adrenoreceptors in rat and the data will help in the understanding the mechanism of action of certain drugs and also desigining new drugs which will become effective where adrenergic neurotransmitter is defective.

Bone disorders encompass a wide spectrum of conditions associated with imbalance of osteociastic and osteobalstic activities. Osteoclasts, the scuplptors of bone operate by producing free radicals acting as chisels. These are responsible for remodeling. Indiscriminate production of free radicals leads to oxidative stress mainly due to lipid peroxidation as seen in our pilot study. The present study probes into the role of antioxidants as a palliative treatment for bone disorders. 50 healthy controls and test group comprising of 198 patients suffering from skeletal disorders like osteoporosis, renal osteodystropy, bone malignancy and fractures underwent baseline assessment of biochemical markers viz. osteoblastic markers serum Alkaline phosphates (ALP), Free Ca++ and Inorganic phosphorus (Pi, osteoclastic marker serum Tartarate rsisitant acid phosphates (TrACP) and Malondialdehyde (MDA) and the antioxidant status serum superoxide dismutase (SOD) and Erythrocyte reduced glutathione (GSH). Each test group was then divided in to gropus A (Evinal 400 mg), B (Celin 500 mg) , C (Evinal + celin ) for antioxidant supplementation for a period of 90 days. The results reveal that there is significant improvement in the biochemical markers serum MDA, serum TrACP, serum ALP and antioxidant status SOD and erythrocyte GSH after antioxidant supplemtntation. Antioxidant vitamins E and C individually or conjointly imoprove the bone status in various skeletal pathologies and hence may serve as cost effective, palliative supplements in additioin to curative treatment.

In asthma, proinflammatory cytokines released by alveolar marcrophages (AM), such as IL-1 beta and TNF- alpha, are known to play a significant role in airway inflammation. The release of these cytokines may be triggered by various extrinsic stimuli (e.g. asthmogens). Exposure of cells to stimulus is known to cause stimulus receptor coupling, activation of transmembrane signalling and phosphorylation of some target proteins of proteins of protein kinase c (PKC), the key regulatory enzyme of the pathway. Phosphorylation changes the conformation of the proteins, which is necessary for several cellular functions. The expression of these cytokines by AM may be associated with the phosphorylation of some of these proteins. based on these presumptions, the studies were conducted on AM of asthmatic patients and healthy subjects to evaluate the expression and release of IL-1 beta and TNF- alpha at mRNA and protein levels by various stimuli viz. asthmogens (histamine, methacholine chloride; MCC), mitogen (LPS), agonists of PKC PMA), antagonists of PKC (sphingosine) and bronchodilator (salbutamol), followed by identification of target proteins of PKC and changes in their phosphorylatioin. The results suggest a significant increase in IL-1 beta and TNF- alpha levels in cell free BAL fluid of asthmatic patients as compared to the healthy subjects without significant change in total member of cells in the BALF. Apparently, the AM of the AM of the asthmatic patients are primed to the triggers of asthma, the presence of which in the milieu may induce thhe early expression and release of these cytokines in comparison to the healthy subjects, leading to the perpeptuatioin and orchestration of inflammation in the airways. The expression of IL- 1 Beta and TNF- alpha by AM seems to be regulated by PKC mediated mechanism. This gets support by the changes caused by these stimuli on phopshorylation of the target proteins of PKC of 120, 66, 35 and 25 kDa, as identified in this study. It may be concluded that in asthma, alveolar macrophages are primed to the triggers and the expression and release of these ctokines by them could be associated with the phophorlation of 120, 66, 35 and 25 kDa proteins through PKC mediated pathway.

Folate deficiency is the common sign of chronic alcoholism. ntestinal malabsorption of the folate is a contributing factro to alchol induced folate deficiency. The aim of the present proposal was to elucidate the mechanism of intestinal malabsorption of folate during chronic alcoholism. Male Wistar rats were fed 1g/Kg body weight ethanol (20% solution) orally for 12 weeks. Intestinal and renal brush border membrane (BBM) vesicles were prepared to study the binding and transport of 3) folic acid. The kinetic studies of the folate transport process revealed the folate transport exhibited saturable kinetics and was pH, Na+, temperature, divalent cation dependent.Chronic ethanol feeding was found to decrease the binding as well as transport the folic acid by altering various kinetic characteristics of the processes and different factors impart diverse effects to these processes at both the intestinal and renal brush border membrane. Thus the process seems to be complex interplay of the diverse biochemical and physiological factors at the absorptive and conservative sites in the mammalian system. Decreased uptake of folate across the intestinal epithelium was associated with a reduced expression of mRNA of reduced folate carrier (RFC). Moreover, the transport was reduced all across the crypt villus axis arger chronic ethanol feeding. Using vrious activators and inhibitors of different singaling pathways it was observed that folate transport is regulated by cAMP dependent protein kinase A. Ethanol also seemed to exert its effect on transport of folate by interfering with his regulatory pathway. In conclusion, ethanol ingestion results in reduced renal & intestinal transport of folate by decreasing the expression of its transporter and interfering with regulation of the transport process.

Objectives of the project were to delineate the role of cell surface carbohydrate epitopes on leukocyte and platelet on one hand and of galactose bingding lectin (galectin-1) of endothelial and underlying cells on the other, in anchoring these cells on blood vessel walls. Also to explore the potential of human erthrocyte membrane glycoproteins as a soruce of glycopeptides and oligoscacchrides that may be useful for inhibiting galectin-1 mediated deposition of blood components on vessel wall. Galectin-1 was purified from human heart tissue by affinity chromatography using lactose-Sepharose. The co purifiction of endogenous glycoproteins along with galectin-1 suggested formation of carbohydrate dependent complexes between the two. It was suggested that when the divalent galectin-1 binds to multivalent endogenous glycoproteins the resulting complexes contain enough suger binding sites to facilitate erythrocyte receptor aggregation during hemagglutination by this complex as well as its attachment to lactose-Sepharose during isoltion of galectin-1 Present results suggest lectin-carbohydrate recognition as primary reason for association of co purified glycoproteins with HHL within intact itssue or after homogenization. Copurified glycoproteins therefore are chosen endogenous complementary glycoproteins for HHL and contain sugar ligands apt for the lectin. Further proof for this is the recognition of coated HPLC separated co purified glycoproteins by purified HHL. In this context the abundance of sialylated as well as free T antigen in copurified glycoproteins points to a preference for T antigen by HHL. Erythrocyte membrane glycopeptides were prepared for use as inhibitors of galectin-1 for preventing tumor spread since some saccharides are easily soluble compared to their membrane glycoproteins precursors. From the point of view of a possible therapeutic potential, erythrocyte derived glycopeptides offer other major advantages such as ease of availability of erythrocytes from discarded blood as well as plasma separation and being non immunogenic in humans.

Sensitivity and specificity of fluorescent labelled antibody absorption (FLA-ABS) and lepromin tests were investigated for early detection of subclinical infection among household contacts of leprosy patients (both paucibacillary and multibacillary). FLA-ABS was found to be very sensitive and specific for the detection of subclinical leprosy infection as FLA-ABS positivity in contacts of both multibacillary and paucibacillary leprosy patients was higher than that of lepromin FLA-ABS positivity was much higher in the contacts of multibacillary leprosy patients than paucibacillary patients. On the basis of FLA-ABS positivity and lepromin reactivity contacts of leprosy patients could be classified into four groups viz. (i) FLA-ABS positive, lepromin negative; (ii) FLA-ABS positive, lepromin positive; (iii) FLA-ABS negative, lepromin positive; and (iv) FLA-ABS negative, lepromin negative. Contacts who were FLA-ABS positive and lepromin negative appeared to be at much higher risk of developing leprosy than the other group of patients as most of the contacts who developed leprosy were from this group. Hence, it can be tentatively concluded that FLA-ABS and lepromin testing help in identifying contacts who are greater risk of developing the disease. Follow up of leprosy patients under treatment indicated that FLA-ABS test remained positive even after clinical subsidence of the disease. FLA-ABS positivity was higher in patients of multibacillary leprosy after subsidence of disease as compared to those with pauci- bacillary leprosy. These findings, however, need to be confirmed by larger studies before any definite conclusion can be drawn regarding the usefulness of FLA-ABS testing in the follow up of treated leprosy patients or the contacts of leprosy patients.

This study was undertaken to develop a method for producing sustained eosinophilia in rhesus monkeys, and to find out its effects on the heart in the presence of specific and non-specific immune enhancement. The effect of sustained eosinophilia after specific deficiency of tryptophan and the possible association between eosinophilia and endomyocardial fibrosis were also studied. Sustainable blood eosinophilia was produced in rhesus monkeys by subcutaneous weekly injections of globular sodium alginate beads coated with coat IgG. Eosinophilia of more than 2000 eosinophils/cubic millimeter was obtained after 12 weeks, the count peaked to about 6000 cells/cubic millimeter at 9 months and this persisted upto 12 months of study. Lymphocyte blast transformation studies carried out at two monthly interval revealed that lymphocytes play an important role in the maintenance of the eosinophilic state as evidenced by increased propensivity to proliferate (as measured by tritiated thymidine incorporation) when challenged with a mitogen (PHA). The highest proliferation of lymphocytes was observed at the end of 12 months suggesting that they were secreting certain factors which helped to sustain the eosinophilia. Persistent blood eosinophilia did not produce any significant histo- pathological changes in the heart. No such changes could be found even when the eosinophilia was associated with non-specific or specific immune enhancement, or tryptophan deficiency.

The study was carried out to type a group of monkeys for D 8/17 B-cell alloantigen: creat a laboratory model of carditis using cross-reactive antigen in genetically defined monkeys: and study the antibodies generated against purified streptococcal carbohydrate and membrane antigens alone and in combination in D 8/17 positive and negative monkeys. Adherence of streptococci on oropharyngeal epithelial cells of monkeys before and after sensitization was also studied and cross-reactive antigen generated in circulating immune complexes during various phases of the disease was identified. Of the 117 monkeys typed for D 8/17 B-cell alloantigen 23(19.66%) were positive. The expression of D 8/17 alloantigen on the cells of the monkeys did not change with time after repeated sensitization of the animals with streptococcal membrane or carbohydrate or a combination of these antigens. Repeated injections of streptococcal membrane and carbohydrate antigens did not change the expression of the oropharyngeal epithelial cell receptors binding to streptococci. High levels of anti-membrane and anticarbohydrate antibodies were seen in the sera of the monkeys injected with the respective antigen confirming a heightened humoral immune response to the streptococcal antigens. Anti- membrane and anti-carbohydrate antibodies persisted in the sera of animals sensitized with these antigens for a long time. Streptococcal antigens were present in the immune complexes of animals repeatedly sensitized with streptococcal antigens. Myocardial granulomas were observed in two and focal myocarditis in one of the four D 8/17 positive monkeys injected with streptococcal membrane antigen. Focal myocarditis was observed in two and edema and thickening of valves in one D 8/17 negative monkey injected with streptococcal membrane. Edema of valve cusps was also observed in two and inflammation and thicke- ning in one monkey injected with streptococcal carbohydrate antigen. Edema of mitral valve was seen in only one 8/17 negative monkey. Focal myocarditis was seen in two and edema in mitral valve inone D8/17 positive monkeys injected with a combination of streptococcal membrane and carbohydrate antigen. these data suggest that experimental myocarditis can be produced by repeated injection of streptococcal antigens in rhesus monkeys. Animal injected with membrane antigen apperead to develop lesions predominantly in the myocardium while those injected with streptococcal carbohydrate mainely in the valves. Animal injected with a combination of streptococcal membrane and carbohydrate antigen failed to show any significant lesion. Only minimal change were seen in the myocardium. It thus appeared that genetically identified positive monkeys develop greater lesion compared to the genetically negative monkeys.

The study was planned to carry out detailed virological, immunological and histological study in 40 patients with dilated cardiomyopathy and to evaluate usefulness of betablocker therapy. The endomyocardial biopsy was done in large number of patients and with its help it was possible to identify patients with inflammatory myocarditis for improvement in the therpeutic approach. The technique in situ hybridization for study of viral antigen in the EMB specimens has been established. Detection of antigen in EMB specimens has been established. Detection of antigen in EMB specimens will help to understand the role of viral infection in causation of dilated cardiomyopathy. This is in the first large study in world literature for evaluation of usefulness of propranolol, a nonselective betablocker, in dilated cardio- myopathy. Though because of certain limitations, a randomized trial could not could be carried out but the observations obtained highly suggest the useful role of propranolol in this disease. Further this also shows that non-selective agents may also be useful. These observations do merit a large multicentre study to evaluate the beneficial effects of propranolol in these seriously sick patients. As such the prognosis in these patient is dismal and curative treatment like cardiac transplant is beyond the reach of our people. Similarly a multicentre study to evaluate the role of immunosuppressive therapy in inflammatory myocarditis is needed. Till today no randomized study is available in literature assessing the role of these modalities in patients with dilated cardiomyopathy.

The study was carried out to assess the role of norepinephrine and isoproterenol in the development of hypertrophy in cultured cardiac myocytes of the rat and to find out whether alpha and beta adrenergic antagonists can modulate or block this hypertrophic response. The action of norepinephrine and isoproterenol on the synthesis of contractile and noncontractile proteins in the cultured cardiac myocytes was also studied. Primary culture preparations of neonatal rat cardiac myocytes obtained from one to three day old rats were utilized for the study. Norepinephrine (NE), an alpha and beta adrenoceptor agonists had a very potent effect in producing hypertrophy of cardiac myocytes at a concentration of 2uM, whereas isoproterenol (a beta adrenoceptor agonist) did not produce any such effect as measured by the total protein content. The protein content of the cells increased markedly with NE (199 pg/cell/ day) whereas the increase was least with isoproterenol (35 pg/cell/day) as compared to control cultures (33 pg/cell/day). The increase in protein con- tent with NE was both time and dose dependent with the maximum response occurring on day 11 with 2um drug concentration. Norepinephrine had a stron stimulatory effect on the synthesis of contrac- tile proteins actin and myosin heavy chain in cultured cardiac myocytes whi- le isoproterenol stimulated mainly the synthesis of noncontractile protein The differential response with adrenoceptor agonists and studies with adrenoceptor antagonists (phentolamine and propranolol) indicated that the hypertrophic effect of NE was mediated by its stimulatory effect on alpha adrenergic receptors; phentolamine, an alpha adrenoceptor antagonist completely blocked the hypertrophic response of NE. It is thus concluded that NE has a potent hypertrophic effect on the cultured cardiac myocytes and this is predominantly mediated by its stimulatory effect on alpha adrenoceptors. The present study will provide an excellent model to study the biological effects of various drugs and pharmacological preparations on living heart muscle cells.

Effect of apolipoprotein E polymorphism on the lipid and lipoprotein levels related to risk for cardiovascular disease. The study was carried out on unrelated individuals of Kshatriya (182) and Mala (38) communities from Andhra Pradesh to identify the apolipoprotein (apo) E genotypes and allele frequencies in these population. Among the Kshatriya, five apo E genotypes (3/3, 3/4, 3/2, 4/2 and 4/4) were observed in males and (3/3, 3/4 and 3/2) in females. But in Mala population only three genotypes (3/3, 3/4 and 3/2) were observed in both the sexes. There was no significant gender differences in the frequencies of apo E genotypes in both the populations as well as between tow population groups. The 4/2 and 4/4 genotypes were found to be less frequent and observed only among 2 individuals each (missing in female Kshatriya and both sexes in Mala). The gene frequencies of the three alleles designated apo Summation 2, Summation 3 and Summation 4 for the Kshatriya (male and female pooled together) were 0.0522, 0.8516 and 0.0962 and for Malas 0.0395, 0.9210 and 0.0395 respectively. Gene frequencies for three alleles for the pooled population of Kshatriya and Malas were 0.0500,0.8636 and 0.0864 respectively. In the apo E genenotypes, the apo E4 homo or heterozygotes showed higher levels of cholesterol compared to other genotypes. The influence of each of the three common apo E alleles (Summation 2, Summation 3, Summation 4) on cholesterol and HDL cholesterol levels was evaluated by the test o f average allelic effects. The Summation 2 and Summation 4 alleles showed lowering and and elevating effects respectively on cholesterol levels. But there was no evidence for a consistent relationship between apo E genotypes and HDL cholesterol. The study also supports the assumption that lower Summation 4 frequency may be associated with lowered risk of cardivascular disease in rural population.

Heart Diseases is a preventable chronic disorder of the heart. It is the serious complication of Rheumatic Fever when it is left untreated. The consequences of the disease include increasing disability, repeated hospitalizations and prmature death. This project was organised by the Jai Vigyan Mission Project on control of RF/RHD. A community based survey with both active (School survey) and passive (RF/RHD registry) reporting modes was adopted for the study. For RF/RHD Registry, all reported cases of RF/RHD in this population of 1368715 were listed and population prevalence estimated. In the school survey 25033 school children were actively screened in a meticulous way (first by the project medical doctors, then by cardiologist and confirmation done by an echocadiogram) to estimate prevalence of RHD among childre3n. Extensive health education campaigns on the disease and preventive measures were carried out in the project area. The prevalence of RHD and incidence of RF in the selected regions of Ernakulam district is among the lowest reported thus far from the developing world. It appears to support the view that improved access to health and antibitics results in a decline in the RF and prevalence of RHD.

In connection with the school sruvey for screening RF/ RHD, a sub study was initiated on the anthropometric and blood pressure measurements of school children. A school based cross sectional survey was done where the height, weight and blood pressure measurements of 21000 students of 5 to 16 years of age were taken. on analysis blood pressure in children shows an increasing trend in relationship with age that continues into adolescence. This relationship is seen in systolic as well as diastolic blood pressure. Blood prressure also shows an increasing trend in relationship with height for all age groups. Diastolic hyopertension predominates in children.

India faces an impending epidemic of cardiovascular diseases. Major concern in this respect to our country is CVD's are more likely to attack adults in their productive years of life. This has a profound and adverse effect on households, families and society. The prevalence of various cardiovascular ailments in adivasi population has been poorly documented. The health care infrastructure for these people is also very limited. A cost effective preventive strategy will need to focus on bringing down the risk factors both in an individual and in the population at large. With this aim this project was initiated in Wayanad District in Kerala which have high proportion of adivasis. Objective is to study the distribution and determinants of the cardiovascular disease freequency in the adivasi population and to create a model for performing disease surveillance in diffucult to reach areas using telemedicine. The project team would visit the adivasi hamlets and screen for cardiovascular diseases. The screening include measurement of height, weight, resting blood pressure, heart rate etc. and cardiac auscultation. Basic demographic data, income source, predominant diet, tobacco consumption and nutrition is also collected. Follow up will be done for patients with risk factors for CVD

This project is conducted as a part of the second phase of the registry component of the Jai Vigyan Mission Mode Project on RF/RHD control. It is a satellite project of the Amrita Institute Medical Sciences. Kochi RF/RHD Registry Project and is being implemented at Wayanad district in Kerala. In this phase the project is extended to areas with a relatively poorer health care infrastructure hence the District of Wayanad has been selected. A community based survey with both active (RF/RHD registry) reporting modes was adopted for the study as in the nodal centre, Kochi. The strategies used in the nodal centre, Kochi has been adopted here, but the passive surveillance is done through the entire health care infrastucture abailable in the district.

Study was undertaken to assess the role of electrochemical gradient of protons in growth and differentiation of Candida albicans, a pathogenic yeast. The results demonstrate that a transient rise in intracellular pH preceded phenotype divergence of C. albicans and the magnitude of change in pHi was greater in the evaginating population destined to diverge as buds when compared to that of the germ tube-forming population. These findings suggest that pHi may be a contributory agent in the differentia- tion of C. albicans. There was a rapid rise in pHi around 135 min. which also coincided with the time of evagination. The transient increase in pHi was followed by a rapid drop. The transition of pHi could be correlated to plasma membrane H+- ATPase activity. The uptake rates of various amino acids e.g. L-proline, L-methionine, L-glutamic acid, L-alanine, L-phenylalanine and glycine varied with the evagination. As compared to other amino acids, the uptake rates of L-methionine were much higher in mycelial form. Results demonstrated that pHi and H+-ATPase play an important role in driving the nutrient transport of C. albicans.

Cis-unsaturated fatty acids are known to lower hypertension both in animals and humans, and it has been attributed to modulation of prostaglandin biosynthesis. In essential hypertension, the activity of cell membrane bound enzymes Na+K+-ATPase has also been shown to be altered. Hence, the influence of prostaglandins and their precursors, cis-unsaturated fatty acids (c-UFAs) on the activity of Na+-K+-ATPase was studied in normal individuals and patients with essential hypertension. It was observed that both n-3 and n-6 fatty acids can either augment or inhibit the activity of normal human leukocyte Na+K+-ATPase and 5'-nucleotidase activities and in leukocytes of patients with essential hypertension in vitro. Both cyclo- oxygenase and lipoxygenase inhibitors did not influence the modifying action of n-3 and n-6 fatty acids on the activity of leukocyte cell membrane bound enzymes. These results suggest that the anti-hypertensive action of cis- unsaturated fatty acids can at least, in part, be due to their action on cell membrane bound enzymes.

An understanding of the biochemical mechanism of action of tumour promoters is fundamental to developing methods and strategies for controll- ing the incidence of cancer in humans. Study was carried out for under- standing and comparing the initial events in the mechanism of action of tumour promoters. A series of tumour promoters like phorbol-12-myristate- 13-acetate (PMA), benzoyl peroxide (BP), mezerein and 3-keto bile acid (KA), with different abilities to promote tumours were examined and their rela- tionship studied in different cells in culture i.e. both normal and trans- formed cells, to see intercellular variations, if any, and to overrule cell specific effects. The cell types studied were mouse fibroblast cells NIH 3T3, human epidermoid carcinoma -HeP2 and A431 cells. The cells were grown to 80% confluency in their respective media and were treated separately with standardized doses of tumour promoters i.e. 25 ng/ml of PMA, 125 ng/ml of mezerein,125 ng/ml of KA and 1 mM BP for varying lengths of time. A stimula- tion of poly ADP ribosyl transferase (ADPRT) activity was found with all the tumour promoters in all the cell types studied as compared to the untreated control cells,the effect being more for the more potent tumour promoters and there were individual variations in time when the maximum effect of these tumour promoters was discernible. The tumour promoter induced accumulation of poly ADP-ribose was accompanied by a concomitant drop in NAD levels. The decrease, however, did not always coincide with the time kinetics or the extent of enzyme stimulation. Addition of 3-amino-benzamide (3AB) an inhibitor of poly ADPRT inhibited the stimulatory effect of these tumour promoters on the enzyme activity. As many tumour promoters induce an oxidative burst and cause membrane mediated damage, the intermediacy of active oxygen species was investigated by studying the effect of antioxidants and antipromoters on the enzyme activity. Superoxide anion levels after promoter treatment were also measured. Pretreatment of the cells in culture with enzyme scavengers of active oxygen like superoxide dismutase (SOD), catalase (CAT) and the antioxidant butylated hydroxy toluene (BHT) showed a decrease in poly ADPRT activity to varying extents with PMA and BP., thereby showing that active oxygen species represent intermediates in this reaction. This was supported by a 2.5 fold increase in superoxide anion level with PMA and BP. Antipromoters also showed inhibition in poly-ADPRT activity in the case of PMA and BP treated cells. In order to verify the tumour promoter induced alteration of intra- cellular calcium in the cell types studied, calcium was measured using the fluorescent indicator method. With this technique a substantial increase in calcium was detected to the extent of 2 to 3 fold with PMA and BP respectively in HeP2 and A431 cells. The replication of DNA in A 431 cells as measured by incorporation of labelled thymidine in these cells was stimulated by 36% and 48% by PMA and BP respectively. However, mezerein showed only 21% increase. The effect of these tumour promoters on the subcellular distribution of protein kinase C (PKC) was also observed at 15 min. and 90 min. post promoter treatment. In NIH 3T3 cells, addition of PMA lead to a transloca- tion of the enzyme to the particulate fraction at 15 min. but the effect decreased at 90 min. probably due to down regulation of the enzyme. Simi- lar effect was seen with mezerein whereas BP showed no change. In A431 cells the basal level of PKC was much lower as compared to NIH 3T3 cells but these cells also responded to the tumour promoters in a similar fashion as NIH 3T3. Any protein changes by these tumour promoters in various subcellular fractions as well as the endogenous phosphorylation of proteins was also studied. In NIH 3T3 cells bands corresponding to Mr 66, 60 and 55 KD in PMA, BP and mezerein treated cells were seen. An additional band corresponding to Mr 19 KD was observed with PMA and mezerein treated cells. The most predominant band observed with all these tumour promoters was the Mr 66 KD band. The other predominant bands were in the Mr range of 29 - 14.5 KD. Phosphorylation pattern of the proteins essentially showed substrates at 116 KD and in the region of histones i.e. Mr 29 - 14.5 KD. Untreated A431 cells, as well as on tumour promoter treatment, essen- tially showed similar changes in the protein and phosphorylation pattern as NIH 3T3 cells. The results show that there is a unifying set of control mechanisms to translate the signal by these promoters studied, into intracellular message to the nucleus and thereby result in altered gene expression.

The aims and major objectives of the study included identification of oncogenes in human cancers by cell transformation, studies on oncogene amplification and expression and study of molecular events of neoplastic conversion and determination of role of oncogenes in cell proliferation. Tumor specimens were transported in cold saline and used for DNA or RNA extraction within 3 hours of removal from the patient. Primary cultures of Swiss embryo fibroblasts were maintained and cell transformation studies were carried out in rat embryo fibroblasts . Attempts were made to assess the extent of possible amplification and expression of oncgenes. For these studies, in vitro random primer labeled 32p-oncogene probes of myc, ras and fos genes were used. The tumor DNAs and RNAs were isolated from tumors of scalp, cheek and pinis. Analysis of tumor DNAs by DNA-DNA and RNA-DNA hybridizations indicated amplification and increased expression of the myc gene in these tumors. However, the ras onco- gene was neither amplified nor expressed increasingly in these tumors. In another set of experiments, the role of oncogenes in cell-cycle progression was determined during the course of exposure of quiescent cells to mitogens. To determine the effect of DMSO on the expression of early growth-response genes in serum-stimulated cells, the levels of mRNAs of IL-6, c-fos genes were quantitated by RNA- DNA hybridization. The results showed that the expression of the proto-onocogene c-myc and that of the cytokine IL-6 were completely blocked by DMSO treatment of cells from the beginning of mitoge- nic stimuli. No inhibition of myc an diL-6 expression was observed when the cells were exposed to DMSO at later stages (after 3 h) of serum-stimulated growth. The serum-induced expression of fos gene was completely abolished by DMSO treatment of MEF during the first 30 min of mitogenic stimuli. DMSO did not have significant effect on PMA-mediated expression of the fos gene. DMSO drastically inhibited serum-induced DNA synthesis, it failed to inhibit DNA synthesis in the presence of PMA and FCS. In order to get insight into the biochemical basis of neoplastic develop- ment, normal rat embryo cultures were co-transfected with oncogenes myc and ras. A 48 KDa secreted protein identifed as plasminogen activator inhibitor showed a drastic decrease upon transformation. A 45 KDa secreted protein identified as DNA synthesis inhibitor also showed a decrease upon transformation.

Study was carried out to probe the molecular pathology of the platelets in hyperaggregatory states, especially in coronary artery disease, in a 2-pronged way. The physical alteration in platelet plasma membrane including measurements of membrane microviscosity and lipid order parameter investigated. Effect of relevant drugs (e.g.propranolol) on these parameters was studied and effect of agonists like thrombin, phorbolsters or calcium ionophores (A 23187) on platelet cell biology and stimulus-response coupling system was investigated including assay of activity of protein kinase C, measurements of intracellular Ca and phosphoinositide turnover. 14 C serotonin uptake and release studies were done to assess level of platelet activity. Platelet aggregation and secretion (of 14C-serotonin) was found to be hightened in acute myocardial infarction. Propranolol suppressed both these responses. Propranolol enhanced anilinonaphthalene sulfonate(ANS) binding to platelet membrane. Scatchard analysis showed an increase in number of bind- ing sites for ANS in propranolol treated platelets.From steady-state fluore- scence polarisation studies using, diphenyl hexatriene(DPH) propranolol was found to decrease the membrane microviscosity in a dose-dependent manner, with a simultaneous fall in lipid order parameter and rise in fusion activation energy for viscosity. On the contrary, platelets from acute myocardial infarction were found to have decreased membrane fluidity, as well as fusion activation energy. Preliminary results showed that propranolol depressed protein kinase C activity in thrombin stimulated platelets but not in platelets stimulated with phorbol esters. Propranolol was, however, found to inhibit activities of phospholipase C and calpain. The results of the study indicated that patients from acute myocardial infarction have altered platelet membrane microviscosity. Propranolol, a drug widely used in cardiac disorders for beta-blocking property, has direct perturbing effect on platelet membrane bilayer, and possibly, on platelet transmembrane sigma transduction.

In recent years, pan masalas have emerged in Indian market as safer alternatives to tobacco chewing. The list of ingredients makes a very strong case against the product and to confirm possible risk associated with pan masala consumption, a detailed project encompassing in vitro animal and human studies was initiated. In vitro experiments included three different parameters and experiments with aqueous and DMSO extracts in presence and absence of the S-9 activation system. The results on frequency of chromosome aberration (CA), sister chromatid exchanges (SCEs) and micronucleated cells (MNC) very clearly showed that treatment of CHO cells with aqueous as well as DMSO extracts of pan masala `plain' variety (PM) and one with tobacco (PM-T), resulted in statistically significant elevation in the levels of all three parameters. Frequency of CA in bone-marrow of mice fed with PM/PM-T at 3 dose levels for 1 to 12 weeks showed insignificant elevation at lower doses and significant elevation at higher doses. Most convincing results were obtained with human studies. With a two issue: three parameter protocol, all three parameters i.e. CA and SCE in blood lymphocytes and MNC in exfoliated buccal mucosa, provided a very strong evidence for genotoxic effects of pan masala consumption.

Study was aimed to understand the molecular mechanisms underlying cardiac hypertrophy and myocardial failure in human hearts to identify the molecular signals which are responsible for the onset and maintenance of cardiac hypertrophy and to correlate the presence, if any, of such molecular factors (hypertrophic factors) with the increase in various cellular events associated with hypertrophic growth. The expression of genes which are specially triggered during early stages of hypertrophy both in lower mammals as well as in human beings has been studied. Enhanced expression of proto oncogenes such as c-fos, c-myc, c-ras and heat shock protein gene HSP70 occur in all human heart tissue samples obtained from patients with ventral septal defect (VSD) or Tetralogy of fallot and artrial septal defect (ASD). This may probably explain the activation of cellular events in myocardial cells during hypertrophy. Also, changes in the expression of myofibrillar protein gene such as MLC2, in the tissue of various cardiac disorders have been presented. The appearance of a high molecular weight protein in the sera of patients with various cardiac anomalies has raised the possibility of identifying the early occurrence of cardiac disorder in human beings. The preliminary studies involving the antibody raised against this cardiac hypertrophic specific protein for the inhibition of hypertrophy development in rats has opened new avenues in the diagnosis of hypertrophy development.

The study was carried out in wistar strain male albino rats to identify the metals which are known to produce either reversible or irreversible impairment in the release of insulin form islets of langerhan and establish a correlation between serum/panereas- metal concentration and the extent of impourment in the insulin secretion from islets. Effeorts were also made to study metals mediated alteration in the transdueion of extr4acellular signals responsible insulin release and investigate the mechanism by which metalions impair the bioregulation ann seeretion of insulin. The administration (intraperitoneal; ip) of nickel chloride (NiCl2: 75 Mue mole/ kg), to rats caused dose dependent rapid and transient rise in blood glucose post administration. The increase in the blood glucose levels levels was accompanied with hyperglucogonemia and marginal increase in insulin leading to a sharp decrease in the insulin-glucagon ratio. The NiCl2 administration also caused diminished glucose tolerance in experimental animials. The response of glucose load in Ni-treated rats was maximum at 0.5 hr. and remained elevated at all subsequent time intervals compared to cotrols receiving either Ni or glucose. The nickel administration also cused significant increase in pancreatic lipid peroxidation at all time intervals with concomitantdecrease in GSH and Zn levels. Significant positive correlation was observed between lipid peroxidation and blood glucose levels. Significant but decreasing correlation was also observed between lipid peroxidation and GSH, lipid peroxidation and Zn and blood glucose and GSH levels. Enhancement in lipid peroxidation, along with the impairment in not-enzymatic defence status including levels of GSH and Zn could be related to Ni-induced oxidative stress. Similar to the elevated patterns of blood glucose and lipid peroxidation, a significant increase in the activity of constitutive nitric oxide synthase (cNOS) in adrenals and brain tissue were observed after Ni administraiton. Contrary to the brain and adrenals, the activity of cNOS was reduced almost to three fold in pancreas. The decrease in cNOS activity in pancrease was accompanied with the significant increase inthe inducible form of the NOS (iNOS) activity. The interplay of NOS in Ni-induced hyperglycemia was further evident by the observation that pretreatment with NG- monomethyl arginine, an inhibitor of NOS, significantly reversed the Ni-enhanced blood glucose levels. To further confirm the role of NO in Ni-induced hyperglycemia, administration of L-arginine, a precursor of No formation, prolonged the Ni-enhanced blood glucose level compared to Ni-treated animals. The role of cNOS in insulin re3lease is well documented, however, the cytotoxic action of iNOS pancreas could thus be playing an importment role in modulating hormone release. These results collectively provide for the probable role of ROS and NO, a key reactive molecule in Ni-induced hyperglycemia.

To raise monoclonal anntibodies against poly ADP-ribose and to use them for immunohisto chemical studies of poly ADP ribosylation in tumour tissues: The study was carried out for in vitro synthesis of the polymer, poly ADP- ribose lench by immunofluorescence analysis in patients of squamous cell tumours and explore the possible role of poly ADP- ribosylation in cancer diognostics with the main objective of understanding the importance of poly ADP-ribosylation in cancer diagnostics and early detection and control of cancer. A polymer of poly ADP-ribose with a chain length of 7 resideres was synthezed with the help of poly ADP-ribose transferase extracted from calf liver. With a method using MBSA two monoclonal antibodies could be produced against this polymer by immunizing Balb C mice with antigen and fusing with SP2/O Ag-14 mouse myeloma cells. The antibodies did not cross react with related compounds, such as DNA, RNA and synthetic nucleic acid hompolymers and ADP-ribose mononer. However they recognized shorted chain as well as long chins polymers of ADP-ribose for the antigen- antibody reaction (3 to 25 residues of ADP-ribose). This antibody when used to measure the poly-ADP-ribose with 3-30 residues of ADP-ribose in squamous cell tumours, clearly demonstrated the presence of poly ADP-ribose in tumors by immunofluorescence. The intnesity of the fluorescence was related to the degree of malignancy.

Action of cisplatinum on some unicellular organisms. A study was conducted to investigate the mode of action of cisplatinum on unicellular ciliate species on order to understand the action of this drug on eukaryotic cells and some aspects of its metabolism. Several different species of genus Stylonchia collected from wild habitat were treated with anticancer drug, cisplatinum in the dose of 150 mue g/ml for 2 h. Studies on macronucleus and micronucleus were carried out to study the mechanism of action of the drug. Different species exhibited variable sensitivity towards the antitumour drug cisplatinum in post treatment resistant cells showed that higher resistant was not due to poor mental uptake; these cells showed comparatively higher uptake; Likewise, the germ line nuclei bound at least 3 times more of platinum when cells were treated in situ. The DNA lesions in the micronucleus caused by extremely low platinum dosages were deterimental to the completion of meiosis and formation of a new functional somatic macronucleus. DNA characterization of platinum damaged micronuclei showed unusual fragmentation to low mol. wt. components. Such damage was irreversible leading to the loss of micronuclei and production of amicronucleate strains. it appeared that micronuclear components like non histone chromosomal proteins facilitate higher platinum binding leading to fragmentation of high mol.wt. micronuclear DNA.

Oncogene transcription factors and extracellular factors. The study was carriedout to determine the role of proto-oncogenes, transcription factors and extracellur factors in the development of cardiac hypertrophy in rats. Efforts were also made to study the mechanism of action of these factors and a protein kinate c (PKC) mediated signal transduction pathway in the induction of cardiac hypertrophy. The 182 KDa serum protein appearing at an earlier stage of hypertrophy was found to be responsible for the induction of protooneogenes and muscle specific genes. The immunopreeipitation studies of nuclear extract indicated that 182 KDa serum protein is synthesized and transloeated into the nucleus within 15 min afeter aortic constrictin. The signal transduction to be muscle specific. The very high level of 182 KDa protein on Ist and 3rd days of neonatal rats also suggested to its hypertrophy specific nature. Activation of PKC within 10 min in 182 KDa protein injected animals along with the activation of proto-oncogenes and transcription factors indicated that the signal transcription pathways provoked by the 182 KDa serum protein could be via PKC mediated pathway. These fidings suggest that the 182 KDa protein synthesized immedicately after the imposition of pressure overload could be the molecular signal for the activation of various events like induction of protooncogenes, transcription factors and muscle specific genes associated with the development of cardiac hypertrophy. The transcription stimulating factor of 45 KDa appeared 3days after the onset of hypertrophy at the time when 182 KDa protein occurs at fairly sufficient levels. This 45 KDa protein probably take over to maintain specific genes resulting in increased muscle protein synthesis to maintain the hypertrophic state.

Gene regulation and expressin of brush border enzymes and transport proteins in animals induced for malabsorption. This study aimed at investigaing the underlying mechanism of malabsorption in male Wistar strain albino rats (80-100g) exposed to chronic alcohol ingestion and giardiasis. The results indicated that Giardia lamblia infection in rats induced malabsorption of glucose and decreased the activities of the brush border disaccharidases and alkaline phosphatases which was primarily due to phosphatase, sucrase and lactase in rat intestine. there across the villus axis in response to giardia infection. chronic ethanol feeding also induced solute malabsorption, which was presumably due to morphological alterations in the intestine. G. lamblia induced idental, changes in mRNA levels of the brush border enzymes and also sodium dependent D-glucose levels in both control and ethanol fed rat intestine.

The activity levels of various DNA plymerases in isolated neuronal and astroglial cells from brain of young (4 days), adult (6 months) and old (>2 years) rats, were assessed using the stragegy based on their differential sensitivities towards various inhibitors such as BuAdATP, BuPhdGTP and dideoxy TTP using activated calf thymus DNA and poly (dA).oligo (Dt)12-18 as template primers. From these results it may be broadly concluded that at all stages studied and in both types of brain cells beta polymerase appears to be the predominant one although other DNA polymerases also seem to be present in detectable amounts. It is known that PCNA activates pol delta but not pol . Western Blot analysis of whole brain bomogenates for assessing the PCNA activity at different ages did not show any difference in the levels with age. Further work could not be continued due non availability of purified PCNA. Since beta polymerase appears to be the predominant polymerase present in adult and aging brain cells, further experiments were designed to examine how this polymerase activity is modulated at different ages, especially its ability to remove a specific 3 end mismatch from a synthetic oligodeoxynucleotide duplex and extend the primer strand to the predicted length. It was observed that on the addition of purified recombinant rat liver B polymerase to the neuronal extracts, the extension activity was restored and the age dependent decrease in activity was not seen. This age dependent primer elogation activity was also studied with unlabelled oligo duplexes incubated under standard conditions with one of the dNTPs radioactive. This study suggests that probably beta polymerase activity is the limiting factor in DNA repair during aging.

Nicotinamide (NA) a naturally occurring vitamin and a protease inhibitor, has been reported to be affective against skin ailments. NA also suppresses the tumor development and cell growth in different model systems in vivo/ in vitro. But NA has limited use as its mode of action is not well documented. A knowledge of the stages of neoplastic development can be exploited to predict the effective approaches to cancer therapy. In order to make the better use of NA as tumor suppressing agent we tried to elucidate the mechanism of action and involvement of stages of tumor development. Topical application of NA suppressed TPA induced mouse skin tumor promotion in two stages tumurigenesis protocol. NA modified TPA altered ODC activity, DNA synthesis, TG activity, endonoclease activity in mouse epidemis. These are the marker events suggests that probably these stages of tumor development are affected by NA. Since NA modified all the critical stages of tumor development altered by tumorigenesis, it becomes a candidate of choice for detailed study of cancer control.

Immunoprophylacic properties of 30kDa secretory protein of M.tuberculosois H37Ra in different adjuvant systems and Immunophenotyping during immunization/ infection (No. 53/2/98-BMS). Six formulations viz. poly lactide-co-glycolide (PGL) microspheres, demethyldioctadecyl ammonium bromide (DDA), liposomes, liposomes containing monphosphoryl lipid A and coated with alum (L-LIPA-AL) or without alum (L-LIPA) were evaluated to promote cell mediated immune responses towards 30 kDa secretory protein of mycobacterium tuberculosis H37Ra in comparison to standard Freund's incomplete adjuvant (FIA). Two adjuvant fromulations of 30kDa L-LIpa-AL and 30kDa-PGL showed maximum reactivity in terms of lymphoproliferation w.rt. other adjuvant fromulations and also exhibited a Th1 shift. Flowcytometry analysis of spleen of immunized animals revealed the capacity of these two adjuvant formulation to activate T cells subjects like CD4 and CD8 T cells. The upregulation of B7 costimulaytory molecules (B7 & B7-2) after immunization further proved the ability of the two vaccin3e formulations to activate antigen presenting cells. The imunostimulatory nature of these two vaccine formulations was also reflected in their capacity to reduce the bacilli load from the lungs of the experimentally infected mice. This study demonstrates PLG and L-LIPA-AL as potent adjuvants for future subunit vaccine development against tuberculosis.

Various DNA repair pathways relevant to brain tissue were examined during development and aging. It was found that Base Excision Repair (BER), a mode or repair to take care of minor base changes, mismatches and small gaps in DNA, is drastically reduced in aging brain cells. During the period of this centre , the locus of defect in these pathways in aging nerve cell has been identified. Thus, supplementation of the neuronal extracts from aging brain with DA- Polymerase beta, DNA ligase along with thymine glycosylase and Apurinic/ apyrimidinic endonuclease in some cases, restored in BER acitvity in vitro. Further, aging neurons were found to show enhanced pol beta activity and BER acitvity following electroporation of pure pol beta. These observations are considered to be of far reaching consequences. However, in the case of Non-Homologous End Joining (NHEJ) mode of repair in brain cells was found to be highly error prone and very feeble in joining blunt and mismatched end while ends while cohesive ends are joined with reasonable efficiency which has decreased with age. Attempts to find the locus of defect in this case have been unsuccessful so far. In a separate study, the presence and the role of two newly discovered DNA Polmerases, the pol lemda, pol mue and Topisomerase II and III in developing and aging brain was studied. Both pol lemda and mue are present in different regions of brain and in isolated neuronal cells. No marked changes in levels of these enzymes were found with age while pol lemda in isolated neurons could be found only in young. DNA topoisomerase were analyzed during development and ageing. Topo I, II and III show a decrease during ageing. Topoisomerase II beta shows significant expression throughtout the developmental stage, whereas Topo II alpha levels decreased in post natal rat pups. The analsis of expression of Topo II alpha while neurons expressing Topo II beta. This data suggest that Topo II alpha playing most significant role in proliferating cells, but Topo I beta may be involved in non-proliferative functions of the cells. One of such function, DNA damage and repair activity is measured in siRNA mediated Topo II alpha and beta knockdown cells, the relsuts showed that Topo II beta along with polymerase beta, Ku70. WRN protein plays important role in peroxide induced Double strand breaks repair. Similarly Topo II beta along with polymerase beta were involved in repair of ENU mediated DNA damage. These studies implicate important role of Topoisomerase II beta and polymerase beta in corrobaration in neurons. hence these enzymes may participate in various DNA repair processess, the decrease of both Topo II beta and polymerase beta ageing may contribute to observed deficiency of DNA repair activity of neurons in ageing.

In the initial part of the study not a single cryptosporium was isolated from 375 stool samples of children. Thus, the original aims were modified to include study of other organism cuasing diarrhoea. A total of 1260 stool samples of children were examined for parasitic, bacterial and viral pathogens with special reference to `Cryptosporidium'. It was observed that in the 200 children admitted to the hospital for diarrohea, none showed evidence of cryptosporidiosis. In the 1060 samples from field areas, 12 (1.13%) were positive for `Cryptosporidium'. Cryptosporidium lead to both acute as well as chronic (persistent) diarrhoea and the clinical features consisted of diarrhoea (91%), abdominal pain (50%), anorexia/vomiting (50%) and fever (25%). The prevalence of `Rota virus' was 15.5% in the hospital group while it was lower (9.2%) in the field area group. Serogrouping revealed that group A subgroup I was the only subgroup present in this area. The incidence of 'Shegella sp.' and 'S. typhimurium' was similar in both the groups of children. 'Each.coli' was the most common bacterial isolate in both the groups. Serotyping of strains from the hospitalized children revealed that EPEC was the commonest group (25.5%). Among the other pathogenic enteric parasites, 'Giardia lamblia' was much more common in the children from rural areas (15.1%), compared to hospital group (3%). The prevalence of 'Entamoeba histolytica' was similar in both the groups (3.4% and 2% respectively).

Strains of E. histolytica were isolated from the patients of amoebiasis under two categories viz Acute ameobic dysentery (AAD) and non-dysenteric amoebic colitis (NDAC). The stool samples positive for E. histolytica were inoculated into Boek and Drbohlav medium for xenic cultures. They were regularly subcultured till profuse growth of each isolate was obtained. From each category the amoebae were picked up, 8 from each isolates. The remaining xenic culture was used for pathogenicity in animals (rats). Ther weanling NIN-Wister strains were taken for the assessment of pathogencity for both xenic cultures (Parent) and clone-cultures. The cloning was done according to method of Srivastvaet. al. (1990). A success rate of 80% 60% and 83% were obtained for clonal-cultures obtained from acute amoebic dysettery (AAD), Non-abscess respectively. The clone cultures along with parent-cultures were tested for pathogenicity in rat caeca. The degree of ulceration was scored according to Neal (1956) scoring method. The smear of feacal speciment was prepared for the positive presence of amoebae. It was observed that there is difference in the degree of ulceration caused by parent-cultures (xenic) and clones derived from them. The zymodema pattern of clone cultures was done by PAGE electrophore- sis based on the isoenzyme pattern of four enzymes viz. phosphoglucomutase (PGM), Hexokinase (HK) Maleic enzyme (ME) and glucose phosphate-isomerase (GPI). The absence of alfa-band and presence of beta-band in PGM along with advance band in HK was taken as marker for pathogenicity. All the 16 clones derived from xenic- cultures were characterized and it was observed that zymodema II is pathohgenic zymodema found in this part of the country. This report is in contrast on other report from North-East India which showed that symodema XIV was the pathognic zymodema for Indian population. The studies were based on zymodema analysis of xenic-cultures obtained from North-East part of the country.

Crude soluble extract (CSE) antigen from Cysticercous celulosae was eva- luated by the enzyme linked immunosorbent assay (ELISA) for the detection of antibodies in serum and cerobrospinal fluid (CSE) samples from patients suspected clinically of neurocysticercosis (NC), neurological disorders other than NC and controls. The results were compared with indirect heamagglutination (IHA) test and matched with retrospective analysis of proven diagnosis of these patients. ELISA and IHA were found to be positive repspectively in 88 and 84 percent of CSF and 92 and 87.2 percent of serum samples from proven NC patients. However, CSE antigen was also found to be reactive in a very high precentage of serum samples from patients other than NC by both ELISA and IHA tests. With CSF samples, IHA test was found to be absolutely specific while by ELISA test CSF antigen was reactive in CSF samples from 5 non-cysticercosis patients, one each suffering from disappearing CT scan lesion, tubercular meningitis (culture negative), chronic meningitis, bening intracranial hypertension and non compressive myelopathy. HOwever, possibility of neurocysticercosis cannot be absolutely ruled out in such patients. Crude soluble extract antigan gave 23 peaks when reacted with homologo- us hyperimmune serum raised in rabbits, by two dimensional immunoelectropho- resis and multiple (25-30) bands with molecular weight ranging from 8 to 2 kDa were observed by sodium dodecyl polyacrylamide gel electrophoresis. The CSE antigen was purified by sephadex G-200 column chromatography and major purified fraction was subjected to western blot analysis using positi- ve serum pool from five surgically confirmed NC patients and-negative serum pool from five normal healthy control subjects. Antigenic fractions with molecular weight of 18, 20, 24 and 65 kDa were found to be reactive with positive serum pool and non-reactive with control serum pool.

The objectives were to study the epidemiological aspects of neuro- cysticercosis and clinical manifestations of the disease in Indian subjects, to evaluate the various modes of treatment viz. praziquantel, palliative (symptomatic) and surgical therapy and to study the dynamics of the immune response in the treatment and prognosis of neurocysticercosis. Within a period of five months from July to November 1989, 15 patients of neurocysticercosis were enrolled in the study. Patients were hospitalised and clinical and CT investigations performed. All cases had repeated stool examination for the presence of ova of Taenia solium. Out of 15 patients, 5 were vegetarians. Of the 10 non-vegetarians, three had never eaten pork, did not eat beef and two had neither eaten pork or beef Four out of 15 patients had close contact with domestic animals, 3 with dogs and one with chickens. None of the patients had direct contact with pigs or cattle. All patients were negative for the presence of ova of T. solium in the faeces inspite of repeated examination. Seizures were the most common presenting feature of which generalised seizures were more frequent than focal or mixed type. Signs and symptoms of raised intracranial tension associated with visual disturbances were also frequently observed. The lesions seen on C.T. scan of the brain were of the following types: (1) Multiple ring like lesions with enhancement on contrast C.T., (2) Multi- ple calcified rice like lesions situated in the brain parenchyma, (3) Iso- dense lesions with extensive white matter oedema and chinked lateral ventricles, (4) Intraventricular enhanced cystic lesions, (5) Granuloma. Specific anticysticercus antibody in serum and CSF was estimated by the ELISA test. Of a total of 12 patients who were tested for the presence of antibodies, a significant antibody response was seen in the serum and CSF in 7 cases each. Symptomatic patients with type (1) and (3) lesion on C.T. scan were given praziquantel 50 mg/kg in three divided doses for 15 days. Decadron 2-3 mg twice a day was also given along with praziquantel. Symptomatic patients with type (2) C.T. lesions and negative immunological response were treated with anticonvulsants alone. Surgical therapy, i.e. removal of cyst was reserved for patients with type (4) lesion on C.T. Evaluation of treatment was based on clinical criteria, CT picture and immunological response. Follow up was inadequate to arrive at definite conclusions. Some preliminary observations which need further validation are as under. The disease was seen in vegetarians and Muslims, thus substantiating the observations by other Indian workers and contrary to well documented studies from other parts of the world who found the condition to be related to eat- ing of undercooked infested pork. Granulomas were seen on C.T. in one case, this has not been reported earlier in Indian patients. Only 60% of cases showed a significant antibody response, one of two cases that worsened after praziquantel and one case that remained the same did not show adequate antibodies either before or after treatment. The possibility of anergy in these cases warrants a study of their immune status. Three cases out of 10 treated with praziquantel were refractory to treat- ment and even worsened. It is not clear whether the patients had an exacer- bation of the disease per se or developed hypersensitivity to praziquantel. Six cases out of 10 treated with praziquantel continued to require anti- convulsants. No relationship was observed between the clinical response and changes in C.T. morphology after praziquantel therapy.

A total of 147 strains of 'aeromonas' of which 54 were isolated from cases of acute diarrhoea and 93 from environmental sources (44 were from water and 49 from fishes) were speciated to 'A.hydrophila' A.sobria' and 'A.caviae' on the basis of only 7 simple biochemical markers, the remaining being common for all of them. 'A.caviae' and 'A.sobria' were predominant in environmental and clinical sources respectively, while the frequency of 'A.hydrophila' was sparce in both the sources, while 56% of the strains produced enterotoxin prior to passage, while the remaining 44% (approx) did so only after 1-3 passages through rabbit gut. The environmental isolates of 'A.hydrophila' and 'A.sobria' produced significantly more enterotoxin (P<0.005) than their clinical counterparts, however, the 'A.caviae' strains did not show such sourcewise difference. Further, 'A.hydrophila' and 'A.sobria' produced significantly more enterotoxin than 'A.caviae'. Nearly 73% of strains produced beta-haemolysin irrespective of their species in the initial experiments, while the remaining did so only after passage. 'A.hydrophila' and 'A.sobria' elaborated relatively more (16-128 HU/ml) haemolysin than 'A.caviae'. Only a small percentage of 'A.sobria' and 'A.caviae' produced alpha-haemolysin. The 31 non- haemolytic strains were distributed into the three species, majority being 'A.caviae'. No correlation could be observed between production of haemolysin type and sources of isolation of the strain. nearly 67% of the beta and 31% of alpha/non- haemolytic strains produced enterotoxin prior to passage, but on passage all of them switched over to production of beta- haemolysin and enterotoxin. Forty four of 97 strains of 3- species of 'aeromonas' caused haemagglutination independent of source of origin. Proportionately more 'A.hydrophila' showed HA than 'A.sobria' and 'A.caviae', whereas equal proportion of clinical and environmental isolates of 3 species showed variety of HA pattern indicate that this property may not be used to assign a isolate to a particular source. Majority strains, mostly showing MS, MFS and MFGS-HA patterns caused accumulation of fluid independent of origin in the initial test. The nontoxic strains, mostly NHA or showed MGS-HA or other patterns because enterotoxin producers after 1-3 consecutive passages through RIL. Twelve strains of 'Aeromonas', 4 each of 'A.hydrophila', 'A.sobria' and 'A.caviae', when tested for adherence to rabbit intestinal mucosa, adhered to independent of species designation or source of isolation. Inhibition of HA or adherence by different sugars indicate that haemagglutinin and adhesin are probably the same molecule. The data thus indicate that all strains of aeromonas whether environmental or clinical in origin are potentially enteropathogenic irrespective of their species. There exists some correlation between HA and enterotoxicity and HA and adhesin.

Electrolyte refluxes in the intestinal mucosa of Vitamin A deficient rats and mice in response of bacterial toxins. The study was carried to determine the Na+, Cl, Ca++ and 3-O- methyl D- glucose fluxes and the key, second merrengers controlling these fluxes in vitamin deficient in male albino mice and Wistar strain male albino rats exposed to different bacterial toxins and to see the reversal of changes, if any after vitamin A supplemenlation. Vitamin A deficiency was produced in animals by feeding them vitamin A deficient diet. This was reflected by cessation in weight gain and confirmed biochemically by reduction in lever and plasma Vitamin A levels compared to controls.There was a significant increase in malondialdehyde levels also the levels of antioxidant enzyme superoxide dismutare in Vitamin A deficient animals thore by making the animals more prone to infection and disease. Both heat labile and heat stable enterotoxins of Escherichia coli decreased the Na+, Cl- and D- glouse absorption and increased Na+ and Cl- secretion in small intestine whereas there was a significant increase in Ca2+ absorption in ST toxin treated animals compared to control. The study related to toxin treatment in vitamin A deficient animals could not be carried out. However, the findings of the study clearly demonstrated that the mucosal barrier of the gastrointestinal tract can be severly impaired by vitamin A deficiency making it more likely to be continously exposed to Gijher level of environmented challange

Control of gene expression in higher organisms is related to the methylation of cytosine in DNA. The pattern of methylation is inherited and is under the control of hormones in hormone dependent tissues. Study has been initiated with the investigate the pattern of methylation and demethylation in steriod hormone dependent and gonadotropic hormone dependent tissues. Such a study would give us information on mechanism of action of hormones at gene level. It is proposed to use two model systems for these studies. One of the systems would be to use sterioid hormone dependent tissue, ventral prostate and uterus. The pattern of methyl transferase and methyl cytosine groups in the DNA following hormonal deprivation and in response to steroid hormones would be studied. In the second system the pattern of methylation of DNA in gonadotropin hormone dependent tissue liketestis will be studied addition to the studied on the regulation of the enzyme DNA-methyl transferase in the gonadotropic hormone dependent tissues. Studies will also be undertaken on the pattern of DNA-methylation in specific ornithine decarboxylase gene of ventral prostate in relation to the levels of hormones.

Measurement of peripheral plasma renin activity and renal vein renin activity has been used to diagnose the potentially curable cases of renovascular hypertension. Plasma renin activity (PRA) is an indirect measure of renin-angiotensin system (RAS) since Ang-II is the main vasoactive harmone responsible for hypertension. Recent evidences have indicated the existence of two types of renins, active and inactive in the plasma the former corresponding to Ang-II. Study was therefore conducted for establishing and standardising the radioimmunoassay and bioassay of Ang-II and evaluating its diagnostic and prognostic value in comparison to plasma renin activity in patients of renovascular and renal parenchymal hypertension. Peripheral Ang-ii levels were estimated in normal subjects and the effect of age , sex, posture and dietary salt was evaluated as also in renal venous blood in patients of renovascular and chronic renal parenchymal hypertension was estimated in some of the samples to correlate the values with Ang-II levels. Significant decrease was observed in PRA and Ang-II levels at higher age in normal males and females. Low salt diet led to significant increase in plasma angiotensin-II accompanied by directionally similar changes in plasma renin activity in normal valunteers. The mean peripheral values for Ang-II were found to be higher in patients of renal parenchymal hypertension. The predective value of renal peripheral vein Ang-II levels for favourable surgical outcome could be evaluated only in patients of renovascular hypertension. Lastly the value obtained by the procedure developed in the laboratory and those obtained by using Ang.II test kit (Buhlmann lab. Switzerland) could not be compared as the kit did not work because of the delay in transit and expiry of the radiolabelled AngII the results of the study reveal that Ang-II and /or PRA measurements are required as supportive tests for the diagnosis of renovascular and renal hypertension. Ang-II can be measured by bioassay as well as by RIA with almost equal sensitivity and specificity however bioassay appeared to be more cost-effective.

The study was carried out on Wister strain albino rats (40 + or - 5g body wt) to investigate the effect of lead exposure (through different routes) on foetal development at various gestational stages in iron defi- cient rats so as to provide information on the fetotoxic and embryotoxic effects of lead exposure. Pregestational lead exposure (1g/1 as lead acetate/nitrate) for 30-60 days through drinking water was found to delay the onset of estrus cycle, more so, in iron-deficient rats. Their mating with normal males resulted in reduced gestational weight gain and enhanced incidence of resorptions probably due to the higher lead uptake in maternal blood and feto-placental unit. Iron deficient diet to rats also affected the blastular hatching in vitro from zona pellucida when lead (10 mu mol/1) was added to the incuba- ting medium. However, iron deficient diet did not seem to affect the impla- ntation sites. Lead intoxication in iron deficient rats also affected organogenesis (6-14 days of gestation) and fetal development (15-20 days gestation). After exposure to lower doses of lead (25 and 50 mg/kg) a dose- dependent feto-placental transfer of lead was seen, it did not produce significant embryo and fetotoxic effects. However higher doses of 100 and 200 mg/kg produced significant dose-dependent impaired gestational development viz increased number of resorptions and reduced litter-size, apart from soft tissue and skeletal deformities. such deformities were more pronounced in iron-dificient lead nitrate treated groups. Intravenous lead administration (single dose; 25mg/kg) on days 9 and 15 of gestation resulted in reduced litter size crown -rump length and fetal weight higher resorptions and abnormalities like hydronephrosis, haemorr- hagic coelomic cavity megacephaly etc in iron deficient groups as compared to those fed normal diet. The 50 mg/kg lead treatment (intravenously) on days 9 and 15 of gestation resulted in higher embryonic deaths (compara- tively more in iron deficent rats). The study thus revealed a dose-dependent feto-placental lead uptake more so in iron-deficient rats reflecting possibility of higher embryo and fetotoxic effects in iron deficiency.

The efficiency of hydroquinone (HQ) and 1,2,4-benzene-triol (BT) iron chelates to act as prooxidants was studied in Wister strain albino rats. Evaluation was done by studying (i) the potential of HQ and BT to release iron from ferritin and whether the iron released could catalyze oxidative reaction; (ii) efficiency of HQ and BT in catalysing bleomycin-dependent DNA degradation, glutathione degradation and lipid peroxidation in vitro; and (iii) the antioxi- dant potentials in serum and liver of rats exposed to benzene. Administration of benzene consecutively for 10 days to rats of either sex resulted in decrease in antioxidant potentials in serum. Serum uric acid and albumin showed significant decrease in all groups exposed to benzene. Alpha-tocopherol in serum and liver did not exhibit significant changecompared to control. Exposure to benzene also led to incrase in lipid peroxi- dation and decrease in free sulph-hydryl groups. Serum ferroxidase activity, total iron content (TIC) and total iron binding capacity (TIBC) measured in female rats exposed to benzene showed significant decrease in ferroxidase activity without any change in TIC or TIBC. The decrease in antioxidation potentials observed may be due to the oxidation reaction exhibited by the benzene metabolites, particularly HQ and BT resulting in oxidative stress in benzene treated rats. Lower concentrations of BT resulted in the release of iron from ferritin which was able to induce lipid peroxidation and catalyze bleomycin-dependent DNA degradation. The presence of oxyradical scavengers ie albumin, catalase or superoxide dismutase significantly inhibited iron release from ferritin by BT. It was also found that iron complexes with HQ and BT catalyzed bleomycin-dependent degradation of DNA. Iron catalyzed bleomycin-dependent degradation of DNA was enhanced 12-fold in the presence of glutathionyl hydroquinone (GHQ) and three-fold in the presence of glutathione (GSH). The degradation of DNA was linear with the increase in concentrations of GHQ or GSH, other factors being constant. The presence of oxyradical scavengers viz thiourea, mannitol, albumin, superoxide dismutase, catalase and dimethyl sulphoxide caused significant inhibition of degradation by GHQ and iron. It is concluded that the poolyphenolic metaabolites of benzene enhance the availability of FE(II) to bleomycin. Such FE (II) was possibly made available due to reduction of FE (III) by polyphenols thus protecting the FE (II) from hydrolysis to hydroxides and further oxidation. It is also concluded that GHQ is a potent prooxidant, capable of enhancing iron dependent formation of super- oxide radical and also due to its faster autooxidation to glutathionyl benzosemiquinone may facilitate formation of adducts with DNA which is one of the prerequisites of benzene toxicity and carcinogenesis.

Ergonomic evaluation of different modes of load carrying maximum permissible load for Indian Women engaged in manual material handling. The study was carried out to document the existing modes of manual load carrying by Indian women and determine the phsiological cost involved during the process. The basic approaches (field and laboratory studies) were employed for the study. Several paramenters like personal and medical history. sites of pain the body, etc. were acertained through a questionnaire. Anthropometry, physiological cost of the work, work rest cycle, walking speed, environmental heat load and energy consumption were determined by direct measurements on the subjects in the field. Women involved in construction activities carried the load on the head shoulder. waist and hand mode were observed. Besides these, women had adopted several other modes for carrying water like head and hand. head and waist. waist and hand etc. Ninety three women engaged in construction activities and 7 women engaged in water carrying activities were selected at random for detailed study. Anthropometric details were recorded on 47 women workers from construction and 25 women workers from water carrying activities. In the case of women engaged in construction activities the average body height, weight and age were 149.7(+-1.7)cm, 42.8(+-5.5)kg and 28.5(+-7.1)yr. respectively. The percentage of fat, lean body weight and body surface area was 21.2(+-3.7)%, 34.0(+-.09)kg and 1.5(+-0.1)m*m, respectively. The average energy intake was 1355 kcal. The average walking speed in the field was 3.4 Km/h and the maximum heart rate of 163 beats/min was observed for women engaged in block carrying. Heart rates during other activities like carrying concrete for column and beam, carrying concrete for block making and mud transferring were between 110-135 beats/min. The O2 demand varied from 0.771/min (45.31% of VO2 max) to 1.411/min(82.7% of VO2 max) being lowest at ground level and maximum after climbing to the 4th floor. The maximum aerobic power obtained was 1.711/min. A regression equation between heart rate and O2 consumption was derived. In case of women carrying water the average body height, weight and age was 150.0(+-5.3) cm. 50.4(+-10.9) kg and 30.5(+-8.1)yr respectively. The fat content lean body weight and body surface area was 25.6(+-5.9)% of the whole body weight as compared to those involved in construction activity. The women involved in water carrying had work experience ranging from a few months to 25 years and more. The water was being carried by HANDA and load varied from 10 to 50kg, while in case of construction activity the weight varied from 9-34kg. Women carrying water reported the back, elbow, upper arm, calf and knee joints asthe major sites of pain. Similar sites of pain were also reported by the construction workers. About 6.5% women water carriers reported miscarriage while miscarriage was reported by 8.3 women involved in constructioin activity. The average walking speed of fro water carrying was 3.8 km/h which was slightly higher than that of construction group. The average working heart rate was 129 beats/min The average maximum heart rage and O2 consumption for women carrying water during maximum aerobic power was 182.3 beats/min and 1.471/min respectively being lower than that of construction workers. This indicates that the physical work capacity of the construction workers was superior to that of women engaged in water carrying activity.

The study was carried out on female Wistar strain albino rats (Wt. 180+20g) to produce ascending pyelonephritis using uropathogenic strain of Escherichia coli, to isolate and purify pili and K-antigens and to find out whether immunization with pili and K-antigens provides protection against ascending pyelonephritis. Ascending pyelonephritis was produced successfully in the rat model by inoculating uropathogenic E.coli 06 into the bladder with and without ligation of the left ureter. Histopathology of the kidneys showed signifi- cant increase in severity score on days 7 and 14 post-infection in both obstructed and unobstructed kidneys. Purified capsular polysaccharide K-antigen from uropathogenic E.coli contained less than one per cent contamination of nucleic acid and protein. Coupling of K-antigen with bovine serum albumin (BSA) produced a high molecular weight complex eluted into void volume of sepharose 6B column. Immunization with two doses of K-antigen-BSA conjugate at four weeks inter- val protected 60 per cent rats from ascending pyelonephritis. However, either K-antigen or BSA alone failed to protect the animals against the disease. SDS-PAGE analysis of purified pili antigen showed a single band at molecular weight of 17 KDa. When tested pili antigen was found to be immunogenic. Animals receiving either active or passive immunization with pili antigen had comparatively low severity score compared to non- immunized infected animals sacrificed on days 7 and 14 post-infection. Pyelonephritis produced damage in the kidney and the transport of glucose and amino acids through renal brush border membrane was also altered (reduced). The changes in the uptake of glucose, L-alanine, L-lysine and L-proline amino acids were partially restored after immunization with pili antigen.

Studies were carried out with the objectives to standardize an animal model for obtaining accelerated fracture healing and a method for confirming the formation of extra callus formation over the control cases (un- stimulated). To understand the mechanism of acclerated fracture healing, experiments were further conducted to measure the strain on bone surface in response to electrical stimulation, and for measurement of strain generated potentials. A new pulsed radio frequency electric field pattern evolved from bone studies was adopted to accelerate fracture healing in rats. The output of a specially designed signal generator was capacitatively coupled to the fracture site using a pair of stainless steel electrodes. In a series of experiments performed on rats, fractures were induced in the femoral shafts and electrical stimulations were applied to one leg. Bone mass formed in the gap was estimated by measurement of the cortical thickness and by ultrasonic attenuation. It was found that the stimulated side showed greater bone mass than the contralateral control. This device offers a simple and reliable method of accelerated healing, immediately after fracture.

A population of about 16010 in filariasis endemic area for around Anji PHC, was surveyed with the help of staff of the National Filariasis Control Programme, Wardha and Community Medicine Department Mahatama Gandhi Institute of Medical Sciences, Wardha. Night blood smear was positive for 731 cases. Thorough personal, family and clinical history and examination was done and out of these 160 cases were having Orthopaedic or surgical manifestation. The patients were classified in 3 groups. Group A - Orthopaedic manifestations of non-filarial etiology. (No signs and symptoms suggestive of filariasis) - 102 could be followed. Group B - Orthopaedic manifestation of probably filarial etiology - 21 could be followed. Group C - Orthopaedic + Surgical manifestation (filarial cases). Serological investigations showed 148 were positive for filarial anti- body. 20 had eosinophilia, 8 were positive for ASO, 4 for CRP, 11 for RA factor and all were negative for sickling, 135 were found to be sterile after culture, 63 patients showed radiological changes. In Group A & B: 57 & 13 patients were treated with either Aspirin, DEC or Aspirin + DEC for 6 months, 79% showed good response to Aspirin + DEC, 58% for Aspirin, 48% to DEC. In group C in 22 patients - synovial biopsy showed non-specific synovitis, 6 were positive for filarial antibody, 2 for RA factor and with chemotherapy none showed response none that patients presenting with orthopaedic manifestations in the area endemic for filariasis test for filariasis should be done for better management of patients. Re-ELISA was negative in 12% in group A, 20% in group B, and none in group C. Hence it is suggested that different orthopaedic manifestations in endemic area based in immunodiagnosis may be helpful for better management by appropriate treatment.

A prospective controlled randomised study was carried out in 48 (35 male and 13 female) patients to determine the role of sodium nitro- prusside (SNP) induced hypotensive anaesthesia, in reducing the operative blood loss and portal pressure during elective proximal lienorenal shunt operations for portal hypertension due to extra hepatic obstruction (EHO) or non-cirrhotic portal fibrosis (NCPF). Twenty five patients received SNP intraoperatively to reduce the mean systolic blood pressure by 26 per cent whereas 23 patients matched for age, sex, body weight and preoperative clinical, biochemical and hematological parameters served as controls (did not receive SNP). Blood loss, the number of units of blood transfused and portal pressure at various stages of the procedure were measured together with the postoperative drainage from the abdominal and chest tubes till they were removed. The portal pressure showed a consistent and significant decrease of 15 per cent with the 26 per cent reduction in the mean systolic pressure. SNP induced hypotension produced a significant decrease in the operative blood loss (mean 701 + OR - 633 ml compared to 1319 + OR - ml; P<0.05) and in the number of units of blood transfused (1.3 + OR - 1.3 compared to 2.17 + OR - 1.10; P<0.05). There was a significant decrease in the operating time from 292 to 265 minutes with the use of SNP. In subjective assessment, the operating field was clearer and procedure was easier in operations where SNP was used. There was no significant alterations in the blood gas measurements, nor were any complications due to SNP infusion. There was no deterioration in the liver function in patients and no alteration in cerebral or renal function during or after surgery in patients who received SNP. In addition, there were no differences in the postoperative drainage from both sites, haemoglobin and haematocrit in both groups of patients. It is concluded that SNP induced hypotension reduces the operative blood loss and transfusion requirements during splenectomy and lineorenal shunt surgery in patients with portal hypertension due to EHO and NCPF. The technique is safe, simple and can be adopted easily by any qualified anaesthesiologist.

Patients with a clinico-radiological diagnosis of chronic calcific pancreatitis (CCP) were studied to evaluate the effect of surgical intervention on pain and the exocrine and endocrine functions. Of the 126 patients (104 males and 22 females; mean age 36.98 years), 40 had alcoholic pancreatitis, 81 tropical calcific pancreatitis, 3 had carcinoma of the pancreas initially presented as chronic pancreatitis and 2 had pancreas divisum. One hundred and twenty patients presented with abdominal pain (of pancreatitis) with an average duration of 4.63 years, 33 had diabetes mellitus, 19 had pancreatic steatorrhoea, 23 obstructive jaundice and 10 patients had leftsided portal hypertension (L-PHT). Pancreatic calcification was observed in 37 of the 126 (29.4%) patients on plain x-ray of the abdomen, while ultrasonography showed dilated pancreatic duct and calcification in 49 (33.6%) patients. Endoscopic retrograde cholangio pancreatography (ERXP) done on 50 patients revealed dilated duct and/or calcifi-cation in all 50 patients. All patients with pain were initially managed conservatively with absti- nence from alcohol, small frequent carbohydrate rich diet, oral pancreatic enzymes and H2 receptor blockers. Sixty nine of the 126 patients not respondingto conservative tretment underwent surgery - modified Puestow's operation in 56,Duval's operation in seven and distal pancreatectomy in six. Internal drainage for symptomatic pancreatic pseudocysts was carried out in 17, biliary enteric anastomosis for obstructive jaundice in 22 and splenectomy for L-PHT in 10 patients. A total of 50 patients could be followed up for periods ranging upto 5 years. Postopertive relief of pain was recorded in 45 (90%) patients. Pain recurred in 4 (8%) patients at the end of the 5 years. Five of the 11 (45.4%) patients with overt diabetes mellitus improved in terms of insu- lin requirement and normalisation of blood glucose values. It is thus concludedthat the drainage operations for CCP do result in a consistent long-term improvement in abdominal pain and diabetes mellitus. Quantitative estimation of inorganic and organic constituents of 8 8 pancratic stones showed that Ca2+ (31.37 + 4.07 mg/dl) and oxalate (5.06+2.18 mg/dl) were the major inorganic constituents and proteins (83.0 + 13.5 ug%) and mucopolysaccharides (6.96 + 1.77 mg%) the major organic constituents.

Using highly sensitive and specific ELISA methods which were developed and standardized, plasma levels of soluble fibronectin(Fn), breakdown product of C3 complement(C3d) and breakdown product of properdin factor B (Ba) were estimated in 64 patients with fulminant hepatic failure(FHF), 29 with subacute hepatic failure(SAHF) and 32 healthy controls without any history of liver diseases. Etiological analysis of patients revealed hepatitis B, hepatitis C and hepatitis NANB-NC infection in 18.8, 42.2 and 39.0 aper cent patients respectively with FHF and 31 34.5 and 34.5 per cent patients respectively with SAHF. None of the patients had hepatitis A infection. Plasma Fn levels were significantly reduced and C3d significantly increased in patients of both groups as compared to healthy individuals, irrespective of viral etiology. However, there was no change in plasma Ba levels as compared to normals levels. Serial estimations of plasma Fn in 19 survivors and in all other patients daily, till death, showed that the rate of rise was significantly more amongst survivors compared to non-survivors. The initial Fn levels were inversely correlated with the aspartate amino transferase, serum alanine amino transferase and prothrombin time in both survivors and non-survivors.

The study was carried out to ascertain the basal status of intracellular free calcium and calcium transport across the plasma membrane in an isolated parietal cell population derived from patients with and without duodenal ulcer. Multiple endoscopic gastric mucosal biopsies from the corpus of the stomachin patients undergoing endoscopic evaluation for epigastric pain/reevaluation ofduodenal ulcer were processed and isolated parietal cells were studied. Gastricbiopsy tissue taken 5 cm away from the lesion in subjects with suspected gastriccarcinoma but having normal acid levels were used as controls. Patients taking calcium antagonists and those concurrently taking H2 antagonists were excluded. Relevant clinical details including smoking habits, alcohol intake, and drug history were recorded. Intracellular calcium was estimated in 21 patients using fura-2-acetoxymethyl ester. Calcium influx was determined in 26 patients and efflux in 12 patients (in 10 patients both influx and efflux could be performed together) by using radioactive calcium. Acridine orange retention was used to assess acid production by parietal cells in 20 patients. A homogeneous population of viable (>75%) parietal cells was obtained with a yield of 1.75x10 cells/ml. The cells had characteristic eosinophilic cytopla-sm and concentric nucleus. The purity of the preparation was demonstrated by a modest three-fold rise in K+-dependent paranitrophenyl phosphate activity from the "crude" to the pure after precoll density gradient. Patients with an active duodenal ulcer had a higher intracellular free calcium (239.75 nmol/10 cells) as compared to patients who had nonspecific endoscopic findings (mean 143.36 nmol/10 cells) and patients with normal endoscopy (mean 145.6 nmol/10 cells). The acridine orange retention as a measure of acid secretion was higher but comparable in patients with active duodenal ulcer (32.55%) and nonspecific endoscopic findings (35.32%) as compared to individualswith normal endoscopy. Only calcium influx at 20 min was significantly more in patients with duodenal ulcers as compared to the control group and those with nonspecific changes at endoscopy. There was no difference between the groupss (with active duodenal ulcer, non-specific changes at endoscopy and normal endoscopy) in calcium influx at 0 and 60 min; calcium efflux at 0,20 and 60 min. None of the parameters tested showed consistent variations that could be attributed to smoking, alcohol intake or presence of nonspecific changes at endoscopy. It is thus concluded that in the unstimulated state calcium homeostasis in isolated parietal cells of pateitns with duodenal ulcer shows only a minimal difference as compared to controls.

Antral mucosal biopsy tissues of 100 patients each of non-ulcers dyspepsia,benign gastric ulcer and with other diseases of gastrointestinal tract were studied for analysing the association of Helicobacter pylori with disease. The biopsy samples obtained on endoscopy from apparently healthy region of the antrum away from the ulcer site served as control. The biopsy samples were subjected to urease tests (indigenously standardised), culture and histopatho- logy. H. pylori was found to be significantly associated in patients with gastritis and gastric, duodenal ulcers and gastric carcinoma. Brain heart infusion agar with 7 per cent horse serum, growth supplements and Butzler antibiotic supplements supported the growth of H.pylori.SDS-PAGE analysis revealed the presence of all the major proteins except the 120kDa protein which was not present in all the isolates. A liquid and a semi- solid urease test standardized indigenously was found to be comparable and cost effective than the commercially available CLO (Campylobactor Like Organism) test. Cytopathic effect was produced by broth-culture filterates of H.pylori confirming the production ofa cytotoxin in HeLa cell line. Intraperitoneal injections of cell sonicates or live suspensions of H. pylori produced lethalityin Balb/c mice. However, injections of heat treated suspension did not produce such effects confirming the production of heat labileby H. pylori. Analysis of data on clinical history of patients failed to reveal the role of alcohol and smoking as associated risk factors in the ulceration process in the patients with H.pylori infection. Irregular dietary habits seemed to play arole in the ulceration process. PCR analysis showed the presence of similar DNA sequences in both Campylobacter jejuni and H.pylori.

Antidelta antibodies were evaluated in 254 HBsAG positive patients to study the correlation of delta infection with severity of liver diseases. The incidence of delta coor super-infection was also estimated using the IgM anti HBc test. Age of patients varied from 1 to 78 years. Of the 254 patients, 148 had acute viral hepatitis and 92 had chronic liver disease (chronic hepatitis, cirrhosis or hepatocellular carcinoma). Eight patients who had received multiple blood transfusion and had no evidence of liver diseases and 6 asymptomatic patients were also HBsAG positive. Antidelta positive was found in 17.3 per cent(44/254) patients. These 44 patients included 23(16.08%) with acute viral hepatitis, 17(18.47%) with chronic liver disease, 3 recipients of multiple blood transfusion and one asymptomatic HBsAG carrier. Patients with delta infection were aged 3 to 63 years. Of the 44 patients with delta infection, 6 gave history of possible parenteral transmission through dental visits, injection etc., 6 had received blood transfusion, 2 were medical professionals and 1 patient had chronic renal failure with hepatitis. Mortality in delta positive patients was found to be 13.63 per cent(6 of 44), while in delta negative patients it was only 8.09 per cent (17 of 210). Of the 44 antidelta antibody positive patients 26(59.09%) had co-infection and 18(40.9%) super-infection. Delta co-infection was seen more commonly in patients wit acute liver disease while super-infection was more common in those with chronic liver disease.Though incidence of encephalopathy as also mortality was higher amongest delta positive patients compared to delta negative patients with HBsAg related acute or chronic liver disease, it was not significant.

Sequential assay of oxygen free radicals, flitrates and citrulline in experimental peritonitis. Sequential production of oxygen free radicals, nitrates and citrulline was studied in New Zealand Wistar strain rats of average weight (200-250 g) after creating peritonitis with caecal perforation. These parameters were also studied after closure of the perforations and peritoneal lavage (ie reversal of peritonitis). Average survival time of rats after induction of peritonitis was 5.33 h. Baseline values of oxygen free radicals, nitrates and citrulline (for comparison) were obtained in sham operated rats (controls) subjected to lpaarotomy and closure. After creating peritontis sequential assays were performed from 1 to 6 h at 1 h intervals. Reversal of peritonitis was carried out at different intervals after induction of peritonitis (from 1-6 h post peritonitis ) and sequential assays were carried out at every 1 h after reversal of peritonitis. There was a steady increase in the value of oxygen free radicals from 0 to 3 h and in the nitrates and citrulline levels from 0 to 4 h, thereafter they showed a gradual fall. Peak levels of oxygen free radicals were seen at 3 h and for nitrates and citrulline at 4 h post peritonitis. Maximum mortality was observed between 3-4 h post peritonitis. After induction of peritonitis all values were higher than the base line values indicating that it was peritonitis which caused the increase. Reversal of peritonitis produced significant fall in all values. This study thus clearly indicated that the values of oxygen free radicals, nitrates and citrulline were significantly increased in peritonitis and were brought down signifiantly with intervention in the from the reversal of peritonitis.

The study was carried out to assess the validity and reproducibility of food frequency questionnaire (FFQ) as a tool for examining the association between various dietery factors and diseases. The study group comprised 100 individuals each ( with no history of illness or modified dietary pattern) from three communities middle income group (MIG), low income group (LIG) and urban slum (US). The FFQ was validated against seven day dietary recall method. The past dietary habit was assessed by using FFQ and data on daily intake for a period of seven consecutive days were collectd utilizing the dietary recall method. The level of agreement between the FFQ and recall was measured by calculating the median difference between the findings from the two methods and the percentage of individuals classified by FFQ to within +/- 1 day per week of the recall. In the reproducibility study the FFQ was administered to a sub sample of indivisuals twice, once at the beginning and again after six months. The median difference between the tow methods for any food item in the data obtained from individuals belonging to LIG was zero. In MIG no median difference was observed in 67% of the food items. The percentage of individuals belonging to LIG, classified by the FFQ to within +/- 1 day per week of frequencies reported in 7 days of recall, ranged from 33.3% for other vegetables to 99.9% for cereals, while those belonging to MIG ranged from 10.3% for other vegetables to 100% for cereals. Similarly individuals belonging to urban slums ranged from 8.6% for fruits to 100% for cerels. In the reproducibility study no significant difference in intake between the tow interviews was observed for most of the food items. The FFQ was found to be fairly valid and reproducible.

The respective study was carried out in 100 elderly patients (aged 60 years and above) suffering from chronic obstructive airway disease to determine the relationship between the family environment and characterstics of the disease (viz. its duration and severity), erception of the famil environment by the patients and caregivers, and medical support services available. The caregivers included sons, daughters, spouses and close relatives. Using semistructured interview schedules significant correlation was observed between the duration and severity of the disease, abailablity of medical support services and patients perception of his/ her family environment. A fair degree of agreement was observed in all the subscales of the family environment scale (FES). In the relationship dimension, high scores was recorded in cohesion and expressiveness indicating to a large extent the commitment, help and support that family members provide to each other and that the faily members are encouraged to act openly and express their feelings directly. Low scores of both patients and caregivers on the issue of family conflict also indicated a supportive family environment. High scores on the independence subscale predicted a tendency to be assertive and self sufficient. The study thus revealed that both the elderly patients and caregivers felt quite positively about their families and preceived their families as supportive characterzed by high cohesion and expressiveness and low conflict associated with family members better adjustment and greater ability to deal with stress, espicially when coping with personal physical illness.

The dietary profiles and nutritional, socio-econimic and health status as also the lhalth behaviour and morbidity profile were assessed in non- institutionalalized elderly (228 men and 205 women) aged 60 years and above. The instruments used in the studyn included a semistructural predesigned and pretested interview schedule to elicit information on the demographic porfilem socio-econimic status self assessment of health status, general health behaviour,morbidity and diaetary profile, anthropometric measurements, blood pressure measurment and estimation of haemoglobin and blood sugar levels. The elderly lived in all categories of women v.z:- high income (HIG), middle income (MIG) and low income (LIG- including slum dwellings). Of the 433 elderly studied, 47(10.8%) alone and 386 (89.2%) lived with the family. Nutritional supplements in the form of vitamins, Ayurvedic and homeopathic preparations were taken by 48,40 and 6% elderly living in HIG, MIG and LIG houses, respectively. A higher prevalenc of smoking and alcohol and tobacco consumption was observed among elderly from the low income group.Alcohol intake, tobacco chewing and smoking has more in men, however, 92% of elderly women from the low income group also smoked. A higher prevalence of diarrhoea, dysentery and respiratory infections was reported by elderly (both men and women) from the low income group. Prevalence of cardiovascular diseases was higher among men compared to women, whereas musculoskeletal problemds, arthritis and pain in weight bearing joint were mone comman in women. Other problems of significance reported by elderly (both men and women) from all strata were cataract and impaired hearing. A greater proportion of women (64%) were anaemic compared to men (46%). The prevalence of anaemia was higher in LIG women (85%) as compared to those from HIG (53%) and MIG(58%). Food beliefs and preferences were strong determinants in selection of food items by the elderly. The men daily intake of all the nutirents showed a decline with increase in age. The iron intake poor across the socio- economic strata, with a deficit of 46% for women and 37% for men. In addition, the intake of protein, calcium and vitamin was alos inadequate in LIG elderly. A gradation was observed in the mean anthropometric measurements with increase in economic status. Among the LIG elderlyn 56% suffered from chronic energy deficiency. Conversely 34% elderly from HIG and MIG suffered from different grades of obesity.

Fibrinolgic activity of blood and effusion fluid in patients with pleural and paritoneal effusion of malignant etiology. The study was carried out in 91 patients with maligrant effusion (4s with pleural and 46 with peritoneal effusion, 33 patients with tubercular effusion (16 with pleural and 17 with periteneal effusion and 26 patients of hepatic currhosis with ascites to investigate the fibrinolytic activity of plasma as well as effusion fluids. Patients with milignant effusion showed a striking lack of plasminogen activator activity in plasma in resulting state. Post-tornique plasma samples, howere, showed release of plasminogen-activator activity in most of the patients. Study on effusion fluid of malignant etiology showed the resemblance of of their fibrinolytic activity with normal plasma behaviour, i.e., lack of fibrin plate lysis by whole fluid, but presence of lysis by the euglobulin fraction. Patients with normalignant etiology showed contrasting results in fibrinolytic activity of both plasma and effusion fluids. In resting state their plasma samples showed similar response as the normal healthy individuals i.e., presence of plasminogen activator in the euglobulin fraction. The effusion fluids of non-milignant etiology also behaved differently from the fluids of malignant etiology. In most of the instance the whole fluid produced lysis on fibrin plate without the need for euglobulin fractionation. Alpha-2 macroglobulin, Alpha-1-antitrypsin and AT-III were assayed in both plasma as well as effusion fluids. The t-PA inhibitor was also tested in effusion fluids. However,no significant increase of these inhibitor was not observed in plasma or effusion fluid samples from cancerous patients as compared to those with having effusion of non-malignant etiology.

Blasts from acute lymphoblastic leukaemia (30 cases), acute myeloblastic leukaemia (20 cases) and chronic granulocytic leukaemia in blast crisis (8 cases) were studied for ultrastructural cytochemistry with the aim to detect lineage infidelity if any, and to detect early differentiation particularly to myeloid lineage. It was also aimed to subtype the leukemia depending on the ultrastructural methodology and cyto- chemical reactions. The samples of blood/bone marrow were collected and after separating the blasts on a Ficoll gradient, they were washed with PBS and processed. Cells were divided into five portions, one for ordinary transmission electron microscopy and the other four for the cytochemical reactions namely myeloperoxidase (MPO), platelet peroxidase (PPO), acid phosphatase and non-specific esterase. Out of the 30 cases of ALL studied, two were found to be AML based on MPO positivity and one case was discovered to be M7 based on PPO positivity. Another 4 cases showed characteristic acid phosphatase reaction indicating the T-cell nature of the blasts. All the cases diagnosed as AML except one case of M5 and two cases of M7 showed MPO positivity in the blasts. One case of M2 and one case of M3 showed more than 10% blasts positive for PPO indicating the megakaryoblastic nature and thus intralineage infidelity. Two cases of M7 showed more than 60% blasts positive for ultrastructural PPO. Of the 8 cases of chronic granulocytic leukemia in blast crisis five were myeloid and three of lymphoid blast crisis. The MPO positivity in myeloid blast crisis ranged from 10 to 90%. One of the cases of lymphoid blast crisis showed 10% PPO positivity. The other two cases of lymphoid blast crisis were negative for all four cytochemical reactions. Thus using MPO, 2 cases of ALL could be reclassified as AML on the basis of ultrastructural MPO positive. In cases of AML and CGL-BC it con- firmed the myeloid nature of the blasts. Therefore, ultrastructural MPO is a specific and early marker of myeloid lineage and of diagnostic importance in unclassified leukaemias. Demonstration of platelet peroxidase (PPO) by ultrastructural cytoche- mistry is one of the specific criterion for diagnosing blasts of megakaryo- cytic lineage. It was found that in two cases of AML a significant number of megakaryoblasts (more than 10%) were there. Two cases of M7 showed more than 60% blasts positive for PPO. Using ultrastructural acid phosphatase reaction, monocytic nature of blasts in AML-M4 and AML-M5 could be confirmed. In cases of ALL, acid phosphatase reaction in golgi region indicated the T-cell nature of the lymphoblasts.

Study was conducted to evaluate the effect of T cell depletion with a specific anti T cell monoclonal antibody and determine whether this treatment can induce haemopoietic recovery in severe aplastic anaemia and to compare this treatment with conventional treatment with oxymethalone with regard to cost, transfusion support required, side effects and response to treatment in severe aplastic anaemia. Patients with severe aplastic anaemia treated with oxymethalone have an 18% chance of response. The response in patients with moderate disease is higher (40%). Haemopoietic recovery was seen in 38% of 34 patients treated with antilymphocyte globulin obtained from MERIEUX. This response is similar to reported results in the Western literature. There was a significant difference in response in patients treated with Hoechst anti lymphocyte globulin (ALG) when compared to Merieux (ALG) and this correlated with the ability of the ALG preparation to induce lymphocyte blast transformation in vitro. This is the first clinical study to document a correlation between response to ALG and its ability to induce lymphocyte blast transformation in vitro. Anti T cell monoclonal antibody WM48 was able to produce profound T cell depletion in patients with acceptable side effects. Treatment of aplastic anaemia with 5 mg of anti T cell monoclonal antibody (WM48) daily for 7 days does not induce haemopoietic recovery, despite profound T cell depletion. Lower dose (1 mg daily for 5 days) of anti T cell monoclonal antibody (WM48) produced a response in 3 of 6 patients treated. In a small number of patients there was a correlation between in vitro lymphocyte blast transformation (LBT) to ALG and haemopoietic recovery with immunosupressive therapy. LBT may be a useful test to identify those patients who are likely to respond to immunosuppression. Data from this study suggest that a combination of lymphocyte depletion and stimulation may be necessary for haemopoietic recovery following immunosupression. Further studies on combination of anti T cell monoclonal antibody with haemopoietic growth factors may provide a more effective therapy and help in the understanding of the biology of aplastic anaemia.

Study was conducted to standardise an assay system for measuring fibronectin levels in serum and the extraction of purified fibronectin from donor blood. To study fibronectin levels in low birth weight neonates with special reference to small for date babies and to study the fibronectin levels in neonates and infants having birth asphyxia and/or systemic infection (pneumonia/sepsis). Extraction of fibronectin from plasma using heparin cold precipitation and its assay by IE and ELISA were standardised. Serum fibronectin was assayed in 6 different categories of healthy and sick infants using immuno-electrophoresis (IE) and ELISA [Term appropriate for date (TAFD); preterm appropriate for date (PTAFD); term small for date (TSFD); preterm small for date (PTSFD); birth asphyxia and sepsis (SEP)]. Term appropriate for date infants were assayed for plasma fibronectin also. Comparison of serum fibronectin levels in different groups by the Wilcoxan Rank Sum test indicated no difference in fibronectin levels between term and preterm infants, between preterm AFD and PTSFD, term SFD vs TAFD, Birth asphyxia vs no birth asphyxia. Septic babies on the other hand had a significantly lower fibronectin level than the non-septic babies. Plasma fibronectin levels in TAFD infants were less than half the levels reported in the western literature for new born term infants. However, adult plasma and serum fibronectin levels in this study were similar to those reported in the literature. No satisfactory explanation is available for this difference. Maternal fibronectin may provide us a clue.

The study was carried out on transfusion dependent patients of thalasaemia for molecular characterization of beta- thalassaemia encentered in Uttar Pradesh. Of the 160 patients of thalassaemia and hemoglobinepathics studied 96 were suffering from thalassaemia major, 36 with thalassaemia haemoglobinepathy, 12 with haemoglobinepathy and 16 with thalassaemia trait. Sixty four of the 96 thalassaemia major patients were hemozygons for one of the 5 common mutations reported from India. Amongst 36 patients with thalassaemia haemoglobinepathy, 24 were of e/Trial 11 of s/trial and one of D/Trial. The DNA samples from a total of 244 unrelated chromosomes from patients with beta- thalassemia and thalhaemoglobinopathy were investigated for 10 (5 common and 5 uncommon) mutations by PCR ARMS technique The prevalence of different mutations was IVS 1,5 (G-->C):60.24%, -619bp deletin:11.06%, Codon 41/42:9.01%, Codon 8/9 : 6.55% IVS 1/1 (G-->T): 3.68%, codon 16:2.46% codon 15: 0.82%and Cap +|:0.41%. 4 cases (1.7%) 2 previously known mutation co30(G-C) and IVS 1-1(G-A) were identified. Though these have been described in Indian samples earlier, there are mainly seen in mediteranean and Middle East countries. Also one new mutation was also characterized. The study thus revealed that beeta- thalassaemia and haemoglobinepathies, particularly HbE and to some extent HbS are important genetic problem in Uttar Pradesh. These disorders are present in the natives of U.P. and in not only those who have migrated from Pakistan. The prevalence of 619 bp deletion is much more common in people of Pakistan origin, while in natives of UP IVS(1,%) G-C mutation was the commonst.

Cerbrovascular acccident is the leading cause of death and is the most common life threatening neurological disorder. The term 'Cerbrovascular accident' referes to all disorders in which area of brain is transiently or permanentaly affected by ischemia or bleeding and or in which one or more blood vassels or brain are primarily impaired pathological processes. In the majourity of cases of peripherial and cerebral vascular diseases, there is a reduction of blood flow to the affected parts of the body. Blood flow properties of blood (Chien, 1993). The treatment of vascular disease is aimed therefore at improving the flow properties in order to produce a flow behaviour compatiable with tissue survival. Haemorheological factors such as red cell concentration, red cell aggregation and fibrinogen concentration were determined in patients of cerebrovascular diseases with assiciated diabetes, hypertension and ischeaemic heart disease. The altered values were observed to be significantly higher in comparision to their controls. The observed values of haemorheological parameters in diabetic and non diabetic parents with reference to normal controls exhibited increasing levels of whole blood viscosity, plasma fibrinogen and red cell deformation with decreased levels of red cell aggeration and packed cell volume.

One hundread ninty normal healthy individuals were screened first for alpha gene deletion, the commonest one accounts the major prevalence among Asian Indian reported by Garewal et al 1994. On screening 125 samples seven were found with one alpha gene defect (4.2%). Two hundred two beta thalassaemia traits were screened for - alpha 3.7 kb, deletion. Out of 202 samples tweenty three were found positive for this deletion (11.5%). A total of 392 samples were screened out of which 31 were found positive for - alpha 3.7 kb deletion (7.9%). Thirty four samples of thalassaemia patients were screened for alpha gene deletion. These thirty four samples include eleven cases of Thalassaemia major and twenty three case of Thalassaemia Intermedia. Out of eleven thalassaemia major cases in two subjects - alpha 3.7 kb deletion was present on both the chromosomes (- alpha 3.7 kb/- alpha 3.7 kb) and one gene deletion was found in two cases (alpha 3.7 kb/aa) indicates overall prevalence of 36% (4/34) in thalassaemia major. In twenty three cases of thalassaemia intermedia one 3.7 kb deletion was observed in three subjects and triplication of alpha gene was found in four subjects. This furtbher indicates the prevalence of alpha gene (13%) in thalassaemia intermedia patients.

The study has high-lighted the critical role the socio-cultural factors play in shaping the health of the community. It is being observed that a vast majority of the respondents are dissatisfied with the health centres located at their plantation. They complain of inadequacy of facilities and the services given by the health staff. Regarding family planning the awareness among the tribal tea garden workers is very poor. It has been observed that the tribal workers of tea plantation have still retained much of their traditional health living in tea plantation for about one hundred years where modern medical treatment and medicine are made available easily to the tribes. However, it has also been observed that in the plantation where better facilities are easily available the tribal workers have shown keen interest in the use of modern medical treatment. But they still believe in their traditional health culture to a large extent. It is, therefore, evident that if proper and adequate medical facilities are provided to the people, they gradually become inclined to accept them. It is often said that deep traditional cultural roots of the tribal is the barrier for accepting modern medical treatment since they have their own traditional magical and herbal systems for curing illness. If modern medical facilities do not reach in the hand of people easily and if modern medical treatment involve such economic pressure, people hesitate to accept them. What we need now for spread of modern treatment is to improve the machinery of health services. Adequate health services, medical staff including doctor, nurse, midwife etc. and medicine are necessary to improve the attitudes of the people towards modern medcine and treatment.

In India, private health care services account for about three fourths of all health care. But information on its utilisation and expenditure is not known in any significant manner. A study was supported at the Foundation for Research in Community Health, Bombay, to explore this important aspect. The only data on health expenditure which is consistently available is the one spent by the central and state Ministries of Health and Family Welfare. Analysis of expenditure has shown to range from 3.2% of total Government expenditure in the early fifties to about 4.3% of Government expenditure at the present. The focus of public health services has over the years shifted to family planning and water supply at the cost of medical care. Thus between 1950-51 and 1984-85 the Government's health expenditure on family planning increased from 0% to 11.4% and on water supply from 1.3% to 23.1% but on medical care it declined from 43.4% to 27.9% of total health expenditure. The private health sector which caters to the latter demand has mushroomed over the years. This is an extremely important finding because it shows that medical care is the weakest link in public health services. In the second part of the study which looked at private household health expenditure in both urban and rural areas, it was found that the average health expenditure borne directly by households is Rs. 182 per capita per year ( in 1987 ), which is 7.6% of per capita total consumption expenditure. This is a substantial amount, especially considering the fact that more than two thirds of the population lives at the subsistence level. A classwise analysis of the data collected in this study shows that with rise in class status morbidity rates increase, use of private health facilities rise, use of public health facilities decline, non utilisation of health facilities decline and per capita health expenditure increases. This means that purchasing power is a crucial factor in health services use and expenditure. Another part of the study has looked into the expenditure and earnings of private medical practitioners in Bombay city. The findings reveal that the average gross earnings of a general practitioner is over Rs. 22,000 per month and their expenditure is about Rs. 4000 per month, giving a net income of Rs. 18000, per month. This study included doctors of allopathy (including licentiates) homeopathy and ayurveda. Finally, the study on corporate sector health benefits shows that the expenditure on medical care expended on employees in the 134 sample companies is Rs. 1339 per employee per year. This expenditure is 2.60% of all emoluments of the employees, 19.33% of all benifits(other than salaries) given to employees and only 0.23% of the companies' sales turnover. On the basis of all these studies it is possible to arrive at a proxy estimate of the total volume of health expenditure in the country. For the year 1989-90, the total health expenditure works out to Rs. 243,500 million which is 6% of GNP. The share of the Ministries of Health is 26% whereas that of households is 60%. Under existing socio-economic conditions this is a highly inequitious situation.

Sister chromatid exchange (SCE) in the lymphocyte chromosomes is used as an indicator of constitutional chromosomal instability in precancerous and cancerous lesions of cervix uteri. SCE frequency was investigated in 100 cases of precancerous lesions of uterine cervix (41 cases of mild dysplasia, 37 cases of moderate dysplasia and 22 cases of severe dysplasia) and 100 cases of uterine cervix cancer. The SCE frequency was also investigated in 150 age and sex matched controls. The analysis of the data revealed that the frequency of SCE varied from cell to cell and from individual to individual. The frequency of SCEs was found to be 7.26 + 1.52 in mild dysplasia, 8.52 + 1.67 in moderate dysplasias, 8.91 + 1.83 in severe dysplasias and 9.68 + 2.19 in patients with uterine cervix cancer. These values were significantly higher than the SCE values in control women. The average rates of SCEs per individual were found to range between 2.56 - 12.14 in controls, 2.95 - 15.32 in dysplasias and 5.16 - 19.26 in cancer patients. The SCE frequencies among the various groups were, however, found to be heterogenous. To investigate whether chromosomes from patients with precancerous and cancerous lesions of cervix uteri are more susceptible to damage in terms of SCE frequency, culture from lymphocytes of some indivi- dual from precancerous and cancerous lesions of cervix uteri were exposed to mitomycin-C (0.01 ug/ml) together with similar exposure to normal age - sex matched controls. MMC induced SCEs were not significantly different from each other indicating that the lymphocyte chromosomes from precancerous or cancerous lesions of uterine cervix are equally susceptible to damage as compared to normal controls and there exists no differential sensitivity. In the present study, the dysplasia cases progressing to cancer showed a slightly elevated SCE frequencies as compared to non- progressed dysplasia cases. The present study thus suggest that an intimate relationship may exist between constitutional chromosomal instability in precancerous and cancerous lesions of cervix uteri indi- cating that SCEs may serve as useful preclinical biological marker in delineating high risk precancerous lesions progressing to cancer from low risk lesions regressing to normalacy.

The assessment of folate sensitive fragile sites on human chromosomes was carried out in 100 mentally retarded individuals alongwith 50 normal subjects. An increase of 8.75% of chromosomal damage was observed in mentally retarded group as compared to the normal individuals. Seven fra-Xq27 individuals have been identified with varying degrees of mental retardation, speech deficit, hyperactivity and autistic features. Of the fragile sites observed in mentally retarded individuals and absent in controls, there were 7 fragile sites which have been reported in earlier literature (i.e., 1q21, 7p13, 11q23, 11q24, 17q12, 18q23, Xq26, Xq27) and fragile sites which have not been reported previously. (i.e., 4p26, 4q33, 7q24, 7q34 and 18q23). Folic acid levels have been estimated in 15 individuals with mental retardation. Of these, two cases had levels below the normal range. They were, however, negative for fra-Xq27. Folate level of one patient identified as positive for fra-Xq27, was found to be within the normal range.

Sister chromatid exchange (SCE) in the lymphocyte chromosome was used as an indicator or constitutional chromosomal instability in precancerous and cancerous lesions of oral cavity. SEC frequency was investigated in 100 cases of oral leukoplakia, 100 case of submucous fibrosis, 100 cases of Palatal keratosis, and 100 cases of squamous cell carcinoma of the oral cavity. Two hundred age and sex matched controls, who has no history of viral infection, hepatitis or any broad spectrum antibiotic therapy at least 4- 6 weeks prior to the collection of blood samples were also investigated. The analysis of the date revealed that the frequency of SCE varied from cell to cell and from individual to individual. The frequency of SCEs (Mean +- S.D) was found to be 8.25 +- 1.74 in oral leukoplakia; 8.91+-1.92 in palatal keratosis; 9.34+-2.21 in submucous fibrosis and 10.23+-2.39 in squamous cell carcinoma of the oral cavity.These values were significantly higher than the SCE values of 5.58+-1.33 observed in normal controls. Combined habits (persons addicated to both tobacco chewing and smoking) induced significantly higher SCEs in lymphocytes as compared to single habits of betel with tobacco chewing or bidi/ cigraette smoking. There was significant correlation between the number of betel leaves with tobacco chewed and the mean SCE frequency and also the number of cigarettes or bidis smoked and the mean SCE frequency in patients with precancerous and cancerous lesions of oral cavity. There was a differential MMC induced SCE induction in pre cancerous and cancerous lesions of oral cavity as compared to normal controls. Stoppage of tobacco usage ( at least for one year)following effective intervention resulted in significant reduction of SCE frequency in patients with pre cancerous lesions of oral cavit. These results indicate that education on tobacco habits should be a feasible and effective method for primary prevention of oral cancer as ceasation of tobacco habits may retard that transformatioin process of normal cells into neoplastic one.

A retrospective and prospective analysis of point mutation of RAS oncogene in patients with myeladysplastic syndrome. The study was carried out on 3 years old bone marrow smear/ metaphase slide preporations of36 myelodysplastic syndrome (MDS) patients and peripheral blood/ bone marrow samples of 19 fresh cases of MDS to investigate the frequency of RAS oncogene mutations (using polymerase chain reaction amplification and differential oligonucleotide hybridization) in order to evaluate the clinical significance of RAS muatations in MDS. Subsequently, clinical information about these patients was also obtained in follow up studies and data were analysed. Among the 36 archival MDS samples studied, 13 were positive for RAS oncogene mutation. Among 19 fresh cases, 6 patients were positive for RAS oncongene mutation. These data, revealed that an increased risk of acute myelogenous leukemia (AML) was found in patients with RAS mutations. Only two patients without RAS mutations. Only two patients without RAS oncogene mutations developed AML. This could be because of the presence of increased bone marrow blasts in these patients. It can thus be on concluded that RAS mutations, although relatively infrequent in MDS, is associated with increased propensity for developing into AML. P. Balakrishna Murthy Department of Toxicology Fredrick institute of plant protection and Toxicology, Padappai

2q 11 microdeletion is the most frequently reported interstitial deletion, occurring in 1/4000 live births. It can result in more than 80 different kinds of birth defects and malformations in varyinhg combinations and varying severity, collectively called 22q11 microdeletion syndromes. Typical facial features, velopharyngial insufficiency, hypoparathyroidism, hypocalcemia and thymic aplasia have been described. Upto 75% of patients with this deletion have been reported to have congenital heart defects, mainly, conoventricular defects. Among patients with conoventricular heart defects, the overall prevalence of 22q11 microdeletion is reported to be between 7% to 19%. The detection and diagnosis of the deletion is of vital importance as it has signigicant bearing on the pre and perioperative management during congenital heart surgery, as well as bearing on the pre and perioperative management during congenital heart surgery, as well as for consuneling regarding other maujor long term issues like developmental delay, immunodeficiency, hypocalcemia, learning disabilities, psychological/ psychiatric disorders etc. The aim of this study is to determine the prrevalence of 22q11 microdeletion among patients with conoventricular heart defects in our population, study the spectrum of phenotypic abnormalities and the correlation between the phenotype and genotype and to study the inheritance patterns. This is a 3 years study which will be completed in september 2009. So far 130 consecutive patients (aged less than 2 years), with conoventricular heart defects, have been recruited into the study. Following detailed cardiac evaluation and classification, these patients are evaluated by a Pediatric Geneticist who describes the phenotypic features. Flourescent In- Situ Hybridization (FISH) (using TUPLE critical region probe) is then performed on blood samples to identify the microdeletion. The preliminary results of the study swhow an overall higher prevalence (21%) of 22q11 microdeletion among Indian patients with vonoventricular derects thanwhat is reported in western literature. The study also sheds new light on the phenotypic features whivh are most predictive for the presence of the microdeletion a combination of thin long fingers, micrognathia and bulbous nasal tip present together being in 90.4% agreement with FISH positivity, on multivariate analysis. This new observation can potentially result in a more accurate early clinical precdiction of the genetic defect, which can help immensely in patient management and counseling. The data requires further validation during the remaining one year of the study.

Systemic connective tissue diseases are among the important chronic disease of young women. There are no data on the prevalence of these diseases from India. Therefore, in the present study, attempt was made to find out the prevalence of one of the major systemic connective tissue disease namely systemic lupus erythematosus (SLE). In the study two types of survey methods were used. In the first method the first-step screening was based upon a questionnaire specially prepared for this purpose. The valididy (specificity and sensitivity) was first established under controlled conditions. Two health visitors were trained adequately to be able to use this questionnaire in the field conditions. These persons screened 39,826 persons using this questionnaire. The population studied was from a semi-rural area of Haryana just bordering Delhi. In the second-step all the persons screened as "probable case" were clinically examined and assessed by a specialist in the field of systemic connective tissue diseases. All the persons screened as "definite case" on the second-step screening, were then also tested for the SLE serum market antibody called antinuclear antibody (ANA). Persons showing definite SLE on clinical examination and having ANA in their serum, were then labelled as SLE. Using this method, the questionnaire based survey showed one SLE case out of the 39,826 persons screened, giving a point prevalence of 0.0251/1000 population; or 25 patients per million of the population. In the second method the first step screening was carried out using a new filter-paper technique for performing ANA test. The technique was thoroughtly validated and also cross-checked by an advanced laboratory for performing ANA test in London. It was also validated by testing a "positive control" (a known patient of SLE in a field area namely Pauri-Gharwal in the hills of Western Uttar Pradesh). After, due validation of the filter-paper technique, 51,361 population from semi-rural and urban areas of Ghaziabad, North Delhi; Green Park are of South Delhi; and a small locality of Pauri-Gharwal, were studied. 13 persons were found to be positive for ANA test. These 13 persons were given a detailed clinical checkup by trained specialists in the field of connective tissue diseases. Their ANA was also repeated using the standard technique. Of these 13 persons, only two women were found to be having definite SLE. Another woman had only some of the features of SLE. She was labeled as "probable" SLE. One additional person, another woman, had an undifferentiated connective tissue disease not having all the features of SLE. The rest of the persons had a "false positive" ANA of no clinical relevance. Using this new method of filter-paper ANA test two definite cases of SLE were detected in a population of 51,361 giving a point prevalence of 0.0195/1000 population; or approximately 39 per million of the population. In the final analysis, it can be concluded that the study detected 3 SLE patients in a population survey of 91,088 persons giving a point- prevalence of 0.0109/1000 population or about 33 cases per million of the population. If one "probable SLE" case and one patient with undifferen- tiated connective tissue disease is also included then the figures will rise to 5.5 per 100,000 or 0.055/1000 of the population. Therefore, it can be concluded that although the disease SLE occurs in the population, mostly in young women, at present its prevalence is not alarming.

The effect of cisplatin on the anti-tumour activities of human NK cells and monocytes was studied. Interaction of cisplatin treated NK cells and monocytes with K562 cells was also studied using the electron microscopy (both TEM & SEM), immunofluorescence (for actin & tubulin) and phase cotnrast microscopy. Mouse monoclonal antibodies against the adhesion or binding molecules or receptors present on NK cells would be raised. Chemiluminescence studies would be carried out on PBML and monocytes on activation and treatment with cisplatin, INF and PMA, and also during their interaction with K562 cells in vitro, with or without cisplatin treatment. The role of calcium ions, microfilaments and microtubules in NK cell/ monocyte - tumour cell interactions in vitro would be studied. Release of tumour cytolytic factors by human monocytes on treatment with cisplatin and their possible separation on FPLC would be studied. Human natural killer (NK) cells and monocytes treated in vitro with cisplatin or rIFN-Y showed enhanced lysis of K562 cells in a time dependent fashion. Cisplatin and rIFN-Y treated monocytes were equally cytotoxic to NK-sensitive (K562) and NK-resistant (Daudi and Raji) cells, whereas NK cells were not rendered cytotoxic against NK-resistant tumor cells. NK and monocyte mediated cytotoxicity against K562 cells was further enhanced when the effector cells were primed with rIFN-Y and then treated with cisplatin. The supernatants collected from cisplatin, LPS, muramyl dipeptide (MDP) or rIFN-Y treated nMNC (with NK activity) showed enhanced release of inter- leukin-1 (IL-1), tumor necrosis factor (TNF) and lysozyme in comparison to untreated nMNC. Treatment of nMNC with cisplatin enhanced the intracellular free calcium levels and protein kinase activity; whereas rIFN-Y treated nMNC showed significant rise in ATP level and protein kinase activity only. Further, cisplatin treated nMNC showed an instantaneous rise in intracellu- lar calcium when NK-sensitive K562 cells were added to the effector cells. Treatment of human monocytes with cisplatin enhanced the expression of membrane bound adhesion molecules, LFA-1 alpha and beta. Addition of anti- LFA-1 alpha and beta antibodies abrogated the monocyte and NK-mediated tumour cell lysis implicating a role of these molecules in the tumoricidal activity of monocytes and NK cells. Lymphokine activated killer (LAK) cells generated in the presence of cisplatin/FK565 along with interleukin-2 (IL-2) showed enhanced binding to tumor cells as compared to LAK cells generated in presence of IL-2 alone. LAK cells showed a reorientation of cellular organelles in the presence of tumor cells. Closer interaction between the LAK cell -target cell and the large number of vesicles in the extracellular space of conjugate pairs may explain the increased lysis of target cells by LAK cells generated in the presence of cisplatin/FK565 along with IL-2 as compared with IL-2 alone.

Study was carried out to see the possible role of the female sex hormone, estradiol, in conjugation with DNA, in the pathogenesis of systemic lupus erythematosus (SLE). Calf thymus DNA fragments (average size 200 bp) were covalently linked to bovine serum albumin and estradiol albumin by treatment with glutaraldehyde. The conuugate was separated from free DNA by gel exclusion chromatography. It was characterised by ultraviolet spectroscopy and thermal denaturation studies. The conjugates induced high titer antibodies which were highly specific towards their respective immunogens. These antibodies did not recognise native DNA (B-form), brominated DNA (Z-form) or poly (rG). poly (dC) (A-form). The estradiol BSA-DNA conjugate showed significant binding with various SLE sera, known to have a high level of anti-native DNA antibodies. The results indicated the recognition of the unique conformation of estradiol- BSA-DNA conjugate by naturally occurring SLE anti-DNA autoantibodies. One of the important findings of this study was the recognition of beta- estradiol by naturally occurring SLE autoantibodies.

The study was carried out in 26 chronic renal failure (CRF) patients (22 male and 4 female,man age of 33 +- 10 yr) and 16 healthy individuals (11 male and 5 females; man age 28.5+-7.7 yr) to investigate the cytokine pathways required for anti HBs antibody production in order to understand the high rate of non responsiveness to the vaccine in CRF patients. The CRF patients received 40 of HBs Ag vaccine at 0,1and 6 months. The relevent cytokines were studied in (i) CRF patients producing protective titres of antibody after vaccination (ii) CRF patients unable to produce prootective antibody titres after vaccination and (iii) individuals showing normal antibody response after vaccination. Man anti HBs antibody titre in normal vaccinated individual was higher (837.3+-236.5 IU/L) as compared to CRF patients (23.43+-236.5 IU/L). Of the 26 vaccinated patients only 13 showed protective anti HBs antibody titres (>=10IU/L) and were referred to as responders while reast of the patients with anti HBs antibody titre less than 10IL/L were non responders.It was found that nonresponder patients had significantly lower levels of IFN-gama in unstimulated cultures, of IL-2 and IL-4 in PHA stimulated cultures and of IL-4 and IFN-gama in HBsAg stimulated cultures suggests that the T cell response in these patients may be skewed to THI phenotype and this is responsible for the low antibody titre

Neurocysticercosis is the commonest parasitic disease of the CNS and commonly presents with seizure disorders. The project aimed at evaluation of Cysticercus fasciolaris, the larval stage of Taenia taeniaeformis, in the immunodiagnosis of human neurocysticereosis. Sera from 50 cases of Neurocysticerosis and 50 healthy as well as diseased controls were evaluated in ELISA using the crude antibodies in case and control sera. Further the sensitivity and specificity of the ELISA was compared using crude extracts of Cysticercus fasciolaris scolex, Cysticercus cellulosae membrane and Cysticercus cellulosae scolex. Sensitivity ranged from 76-94% heighest for membrane sonicate of Cysticercus fasciolaris. Specificity ranged from 78 to 90%. The antigenic profile of all four antigenic fractions was assessed by western bolt with 10 patient sera and two healthy contros. Antihuman IgG or IgM was used as secondary antibody. Several immunoreactive bands were detected between 18-200 KD. Interestingly a prominent band at approximately 66 KD was reactive to both IgG and IgM antibodies in all patient sera tested to memtbrane and scolex sonicates of both Cysticerus fasciolaris and Cysticercus cellulosae. A 41 KD band showed was reactive of IgG antibodies in all cases but reacted to IgM antibody in 6 cases. Further low molecular weight bands at approx. 18 KD were reeactive to IgG but not IgM. The high molecular weight region of >100 KD showed several fineimmunoreactive bands. The 66 KD antigen appears the ideal candidate for analysis in a purified protein based ELISA or Dot blot eassay to eliminate the false positives obtained with the crude extract. Since the immunoreactive bands are common to Cysticercus fasciolaris and Cysticercus cellulosae the C fasciolaris antigen would be good candidate for use in immunoassays since the labroatory development of the larval stages can be carried out in a controlled manner with greater ease and cost effectively in rats.

The study was conducted with the prime objective of isolating an antigen from the crude preparation of whole promastigotes (Leishmania dono- vani) with a view for future exploitation in serodiagnosis and production of monoclonal antibodies. Soluble antigen, prepared from promastigotes isola- ted from a actively growing culture, was fractionated by gel filtration chromatography over a column os Sephadex G200. Three peaks of proteins could be recovered. The antigenic reactivity of different fractions was checked against sera of kala azar patients by immunoelectrophoresis (IEP), rocket immunoelectrophoresis (RIEP) and enzyme-linked immunosorbent assay (ELISA). The first peak was found to be highly reactive in comparison with other peaks. SDS-PAGE and western blot analysis of this antigen revealed a major antigen of promastigotes in the vicinity of 65 to 66 KDa, while a weakly reactive triplet could also be detected in the low molecular weight region.

One of the major requirements of a drug for eliciting leishmanicidal activity is its ability to enter the marcrophage system providing shelter to the amastigotes. The desired molecule chould also be able to interfere with the enzymes system such as superoxide dismutase and glutathione reductase etc. of the parasite to achieve the above biological profile. A series of 5-methyl-4 substituted pyrazoles: 1. 4- substituted dihydroquinolines 2. spiro-naphthalenes 3. pentasubstituted pyrroles 4. and 4-phenyl-1- (b-substituted ethyl) imidazolidin-2-ones 5. and benopyran (3,4-b pyrroles (6) have been designed and synthesised. Most of the compounds synthesised were evaluated against 'Leishmania donovani' in hamsters ('Mesocricetus auretus'). Phytoinvestigation of 'Nyctanthes arbortristis' seeds (F.verbenaceae) have also been carried out and its active principles arbotristocides A-F have been isolated. The results show that new structural leads could be generated. In all 31 compounds were evaluated for their 'in-vitro' leishmanicidal activity. Most of the compounds tested caused >75% inhibition of the amastigotes of 'L.donovani' at a concentration of 100 ug/ml. However, none was able to bring out 100% inhibition of the parasite. A total of 78 compounds were evaluated for their antileishmanicidal activity against 'L.donovani' infection in hamsters. Most of the compounds tested were found to be inactive against 'L.donovani' except compound Nos. 29,30 and 34 which caused 72-95% inhibition of parasitaemia upto day 7 post-treatment. Of these compound No.29 was followed upto day 28 which exhibited 94% activity. Two structural prototypes were designed and synthesised namely substituted pyrazoles and imidazolidones. The in-vitro and in-vivo techniques for evaluating leishmanicidal activity of compounds have been established and a number of compounds have been tested using above test methods. Immunomodulatory activity (prophylactic) studies have been standardised and method to improve drug activity by better drug delivery system has been suggested.

In the present study, a major 66 kD plasma membrane associated molecule of promastigotes of 'Leishmania donovani' (UR-6) has been isolated and affinity purified. The monospecific antibodies were raised against this antigen. The immunoreactivity of 66 kD molecule was lost upon expsure to heat and treatment with trypsin. The metaperiodate oxidation also reduced its immunoreactivity. The 66 kD molecule of promastigotes of 'L.donovani' was, therefore, glycoprotein in nature. Employing a fluorescent probe, the 66 kD molecule was found to be located on the tip of flagellum and on the kinetoplast of promastigote of 'L.donovani'. The affinity purified 66 kD molecule provded partial protection resulting in about 40% reduction in the liver parasitaemic load as compared to unprotected animals. This study thus highlights that promastigotes of 'L.donovani' utilises 66 kD molecule not only for recognising the ligand attached to macrophages also appears to reduce the parasite burden upon immunostimulation of the host.

Study was carried out to characterize the 63 Kd and 54 Kd glycoproteins of the L. donovani promastigotes, determine the antigenicity and immunogenicity of these glycoproteins and to evaluate the role of these glycoprotein antigens in the induction of protective immunity against visceral leishmaniasis in experimental animals. The crude soluble antigen (CSA) of Leishmania donovani promastigotes was shown to contain several proteins in the subunit molecular weight range of about 14 to 100 Kd. Out of these a glycoprotein of 63 Kd was major component followed by another glycoprotein of about 54 Kd. Additional bands in the region of 45, 30-28 and 20-18 Kd were also evident. Sera from kala-azar patients reacted to CSA in the enzyme-linked immunosorbent assay (ELISA) and recognized the 63 Kd component in immunoblots. Some of the sera also recognized 54 Kd, 28 Kd and other bands. Immunization of animals with the CSA or separated 63/54 Kd glycoproteins induced the formation of circulating antibodies against both these antigens detectable by ELISA as well as immunoblotting experiments. Further studies showed that both the glycoproteins had the ability to induce cell-mediated immune response in immunised animals as evaluated by the lymphocyte transformation tests in vitro. Protection experiments carried out in hamsters demonstrated that immunization with 63/54 Kd antigens could induce significant reduction in the parasitic load in vaccinated animals following challenge infection with L. donovani.

Standardisation of DNA-DNA hybridization by dot-blot method using Leishmania donovani promastigotes (UR 6 & GD Strains) has been done. L.D. promastigotes and standard kDNA (both from cultured UR-6 strain) showed positive hybridization spots. Isolation and purification of kDNA from culture of promastigotes (L.donovani) maintained in the laboratory from Kala-azar patients was also performed. For further work it is being stored in the laboratory. On the basis of total protein profile in SDS-polyacrylamide gel electro- phoresis, it was possible to differentiate between the responsive and non-responsive strains of 'L.donovani'. Protein bands of 'L.donovani' and 'P.argentipes' differ. A simple method for crypreservation of 'L.donovani' promastigotes and a medium for its isolation and cultivation without blood have been develop- ed. It was possible to maintain the colony of 'P.argentipes' upto 6th generation. In the biotopes, sandflies (P.argentipes) were found above 1.8m (6 ft.) of the ground level, requiring spray upto ceiling. In a study of population dynamics it was estimated that from 2407 flies collec- ted, a total of 7369 may emerged in a cowshed. Artificial feeding of about 800 sandflies through 6 different membranes were tried for 37 times. In three attempts from chick membrane only, altogether 4 sandflies took artificial feeding mixed with blood and promastigotes. Out of 648 sandflies fed on confirmed Kala-azar patients, promastigotes were found to develop in the gut of only 1 sandfly.

The ability of transfer factor (TF) to reverse the anergy seen in lepromatous leprosy (LL) patients was investigated. 6 active, bacilli- ferous, anergic, LL patients and 6 inactive, nonbacilliferous, treated but still anergic, LL patients were randomly allocated to test and control arms. TF prepared from healthy donors, reactive in vivo and in vitro to M. leprase, was injected subcutaneously into patients in the test arm thrice a week for 6 months (a total of 1.3x10^10 lymphocyte equivalents). Patients in the control arm recieved the diluent, sterile water for injection as placebo, via the same route. All patients were on concomitant appropriate chemotherapy excluding clofazimine. Before starting the trial TF was shown to be free of nonspecific toxicities, and immunologic activity was demonstrated by the transfer of KLH sensitivity to naive recipients. At the end of the trial, only one active LL patient, who recieved TF, showed rapid bacterial clearance from skin and histopathologic evidence of cell-mediated immune response in the lepromin skin test site and in a skin lesion. All other patients in the test and control arms showed no difference The inconsistent effects of TF preclude use as an immunotherapeutic agent in lepromatous leprosy.

Study was carried out with the objectives to investigate if the inheritance of the susceptibility or resistance to leprosy HLA-linked; to determine if there is any association of HLA antigens with different types of leprosy; to investigate families with multiple cases involving parents and/or siblings to establish the sharing of any HLA haplotypes linked with the disease; to investigate the association of HLA antigens with immunity to leprosy among contacts, as judged by the lepromin test and to follow up the above contacts in order to find out whether specific HLA antigens have any bearing on the development of the disease. Study was conducted in 500-600 families to cover about 2000 participants in order to find out the possible association of HLA antigens with the susceptibility or resistance to the disease. Results of the study revealed that none of the HLA-A,-B,-C antigen had any significant correlation with the disease types except in BB group only. In this group HLA-A10 specificity showed a significant association even after correction of the P value. However, the number of BB cases were too small. Unlike earlier reports from the group of deVries results of the study did not show any significant correlation of DR-antigens with any of the disease types. This discrepancy may be due to the inclusion of larger number (408) of families as compared to the study conducted by deVries et al in 1976 and 1980 in which 16 tuberculoid families and 15 tuberculoid patients were selected respectively. Maximum RR values for haplotype frequencies for HLA-A,-B,-C and -DR antigen were noted as follows:- For TT (A26, B8; A29, C2; B22, C2; A30, DR3; B27, DR; C2, DR3); for BT (A25, B21, A2, C1 and A2, C5; B18, C2; A28, DR8; B27, DR4; C2, DR4); for TT+BT (A32, B49; A29, C2; B22, C2; A28, DR8; B27, DR4; C2, DR3); for BB (RR=0 for any haplotype association however, for one haplotypes frequency, i.e. B40, C5 a X2 value of 4.55 (P<.05) was also observed to have a O RR value) and for LL (RR=0 for any haplotype association. However, for three haplotype frequencies i.e. A3, B14; A11, C3 and C7, DR10 having X2 values of 10.90 (P<0.05), 3.92 (P<0.15) and 4.40 (P<0.05) respectively also showed 0 value for RR); in LL frequency of Gc 1-1 was less and Gc 2-1 was more than that of the normals. In LL Gc 1-1 was significantly less and Gc 2-1 was significantly more than those of TT patients.

The study was successful in in-vitro production of T cell lines/clones which recognise the recombinant protein LSR2 as well as the native bacillus. It would appear that even at the level of a T cell line or clone, those T cells that recognise M.leprae are able to proliferate after LSR2 stimulation. Most T cell lines/clones appear to be of CD4 phenotype, indicating that the helper/inducer cells may be selected by the in vitro culture conditions. This appeared to be true for lesional cells also, even though in situ studies had indicated that CD4+ and CD8+ cells are present in BT and ENL lesions. All T cell lines/clones obtained to date showed a/B T cell receptor bearing cells. It had recently been shown that T cells in reactional leprosy lesion had Y/& T cell receptors and it would be relevant to clone lesional cells from more reactional patients. In summary, it is considered possible to address tha question of T cell epitopes at molecular level now that we are able to obtain homogenous T cells and have a recombinant protein whose aminoacid sequence is known. Thus peptides can be synthesised based on amphipathic regions, or as overlapping sequences to define the epitopes required in T cell help. This approach may help in identifying epitopes responsible for immunoregulatory events during the disease process as well as peptides of immunoprophylacti use thus paring the way for second generation vaccines. the study has completed its planned duration.

Studies were mainly conducted to assess the role of the lymphokine IFN gamma to reverse the energy in LL patients and to act as an adjunct to chemotherapy for the control of the disease. The single most significant finding was the reduction in bacillary load in 2/3 of patients. This reduction indicated not only bacillary killing but a clearance of bacilli from the lesions. For ethical reassons the patients continued to receive multidrug therapy and the lesions injected with the lymphokine were compared to those injected with excipient. MDT is expected to reduce the bacillary index by one log over a one year period. In many patients this was achieved within one month and persisted upto 3 months. Further follow-up upto 12 months showed no difference in the bacillary load of test and control lesions. The delayed type local response indicated that 3 consecutive injections were sufficient. Further, increase even after a test period led to refractoriness. There was no significant difference between 10 and 20 mg dose. 8 patients did not show any response even after 3-5 injections of IFN-r. Immunologically, the injected sites showed a migration of CD4+ helper/inducer subset of T cells into the lesions, these my release more IFN-g locally and perpetuate the macrophage activating functions and induction of MHC class II antigens and IP-IO protein. That lesional cells of chronic granulomas of LL patients can be stimulated in vitro to release free oxygen radicals indicates further that IFN-r may be a functionally useful lymphokine in controlling bacilli in lepromatous leprosy. This is perhaps the first report where macrophages from LL granulomas were assessed for functional ability to release H-2 O-2 and O-2.

Nutritional studies on leprosy spectrum including anthropometric measurements, serum albumin, transferrin, vitamin A, vitamin E, serum elements like zinc, calcium, copper, magnesium and iron were determined. Our study has showed that the disease process has no bearing on the anthropometric indices but it has a definite correlation with micro and maccronutrients. Thus we had earlier concluded that leprosy patients had mild to moderate lowering of body measurements, this was due to associated poverty and not disease per se. Data on analysis of micronutrient concentration showed that copper increased from BT to LL end while calcium decreased from TT to LL and magnesium levels increased from TT to LL end. But serum zinc level showed no bearing on the spectrum, though its level decreased in the whole spectrum as compared to normal controls. Serum levels of vitamin A and E progressively increased from LL to BT . The levels of dietdependent protiens in serum samples ( such as serum albumin, transferrin, and prealbumin ) increased from LL to TT end. Serum iron level also increased from LL to TT end. It is tempting to postulate that gliding down by the leprosy spectrum might depend on macro and micro- nutritional markers, however ther is no experiment or clinical evidence to prove this. Growth and development, both physical and psychological of 182 children of leprosy patients were investigated. All belonged to a low socioeconomic status and were born in leprosy colonies to either one or both parents suffering from leprosy. These children were given prophylactic Dapsone monotherapy and administered BCG vaccination. Also 271 control children ( 117 high and 154 low socioeconomic status ) were included for comparison. Body measurements showed that as the boys grew older, body weights, heights and skinfold thickness fell behind in the children of leprosy patients and control children of low economic status as compared to the healthy children of high economic status. Similarly with increasing age, Quetelet indices of the male children of leprosy patients fell behind that of the normal children of the high economic status. Our findings of the body measurements of female children of leprosy patients were found to be similar to that of the male children. As the girls grew older, the height, weight and quetelet Indices also fell behind in the female children of leprosy patients of low economic status, as compared to the normal girls of high socio-economic status. More interest- ingly, the growth of normal female children of low economic status was found to fall behind even to that observed in the female children of leprosy patients. This indicates that the decreased growth velocity of the latter children in relation to the children of high social status was not associated with the illness of their parents, but perhaps due to their economic disparities. The mid upperarm circumference and skinfold thickness of the girls of the high economic status was more than that found in female children of lepromatous patients and normal children of low economic status. The mean haemoglobin levels in boys and girls, normal as well as those of leprosy patients was below 11gm\dl, suggestive of anaemia.

A total of 70 clinicaly early, unclassifiable and doubtful cases of leprosy with disease of less than 12 months duration and with upto five lesions were studied to elicit fine structural changes in early lesions of leprosy, to identify definite criteria for early diagnosis and understand the pathological mechanism involved in the development of early lesions in the leprosy. All patients were subjected to skin biopsy. Light microscopy revealed histologically developed granulomas in 18, indeterminate leprosy in 14 and no lepromatous changes in 38 subjects. Electron microscopy in 25 subjects who had no specific changes on light microscopy, showed nonspecific inflammatory changes in 20 patients. Five patients revealed derangement in the fine structure of the dermal nerve twigs in the form of oedema of the nerve fiber with separation of Schwaan cells and swelling of the axons and splitting of myelin sheath. In addition, fragments of electron dense material which could be of bacillary origin were also found within the Schwann cell cytoplasm. It is concluded that as the electron microscopy of patients with negative histological finding on light microscopy showed specific changes of leprosy in only 20 per cent of subjects, it is not of significant use to confirm the disease.

Aharent cells of peripheral blood mononuclear cells (PBMC) from leprosy patients were stimulated with Phylo Haem Agglutorium (PHA) and Phobal Myristate Acetate (PMA)/M.leprae and the quantity of Interleukin-1 B produced was quantitated by a commercial Elisa kit. PBMC were also stimulated with PHA and PMA/M.leprae and the quantity of the Interleukin-1 B produced was quantitated by a commercial Elisa kit. PBMC were also stimulated with PHA and PMA/M.leprae and the quantity of Interleukin-2 produced was measured by a commercial Elisa kit. Before commencement of therapy LL/BL patients produce significantly lesser quantities of both IL-1 Beta and IL-2 when compared to health control . After 6 and 12 months of multidrug therapy both these levels do not change significantly from their respective pre-treatment level. Before treatment indeterminate and TT/BT patients produce significantly lesser quantities of IL-1 Beta when compared to healthy controls. Like LL/BL patients these levels at the end of 6 months do not change significantly from their respective pretreatment values. Before treatment indeterminate patients produce significantly lesser IL-2 when compared to healthy controls. But TT/BT patients produce IL-2 which is almost same as that of healthy persons. At the end of 6 months of multidrug therapy both Indeterminate and TT/BT patients produce significantly increased quantities of IL-2 when compared to their respective pre-treatment levels. CONCLUSIONS: a) Therefore that a) there is some immunological defect with respect to IL-1 beta production in all the 3 groups of leprosy patients ( in indeterminate, TT/BT and LL/BL). this defect do not change after treatment with multidrugs for 6 months (in indeterminate and TT/BT or 12 months (in LL/BL). b) LL/BL patients also show defective IL-2 production and this defect do not seen to alter after 12 months of multidrug therapy. c) TT/BL patients do not show defectives IL-2 production and after 6 months of multidrug therapy the IL-2 production rises significantly from its pre-treatment level. d) Though the Indeterminate patients show an initial defective IL-2 production, the IL-2 level raises significantly after 6 months of therapy.

The study was carried out on 66 patients with polar lepromatous leprosy or borderline lepromatous leprosy to find out the efficacy of intraderma recombi- nant interferon gamma (rIFN- ) and to evaluate the duration, dose and multipli- city of injections required to induce improvement in clinical and histological features and bacillary clearance. The cellular infiltrates were also studied in the biopsies using immunological markers. After completion of interferon treatment i.e. 4-6 days later, the patients were given multidrug therapy (MDT). The MDT was continued throughout the follow up period. Patients in reaction were excluded from the study. rIFN-y in 100 ul of excipient was injected intrader- mally into well characetised lepromatous lesions. Contralateral lesions, clinically similar in terms of infiltration and erythema were injected with a similar volume of excipient only as control. rIFN-y proved to have potent effects on the lepromatous dermal lesions. Ithad multiple effects, inducing inflammatory responses akin to delayed type hypersensitivity reaction associated with cell mediated immunity. The leproma- tous skin was found to be capable of mounting an immuno-inflammatory response. There was no defect in migration of cells nor an irreversible abnormality in themicroenvironment of the bacilliferous granuloma. The studies involving histology,immunochemistry and ultrastructure gave credannce to the fact that multiple cellular and molecular events were initiated by rIFN- . The skin induration was due to epidermal thickening resul- ting from keratinocyte proliferation and infiltration of T cells and young mono-cytes into the dermis. The predominant lymphocyte was CD4+. Major histocompa- tibility complex (MHC) class 2 antigensnot only increased on macrophages but they were newly induced on hitherto nega-tive keratinocytes. In addition, IP 10,a specific protein induced by IFN-y was also seen in the skin. Repeated injec- tions of rIFN-y resulted in a distinct reduction at 3 weeks of the cutaneous load of organism from 4+to5+to1+to3+. Thedisposal of bacilli occurred faster in the diffused site compared to nodular sites. The immune induced elimination wasrapid and more effective than that observed by anti-leprosy drugs alone. It was associated with activation of macrophages. The study thus indicated that rIFN-y is a powerful biological therapeutic for leprosy. Both improvement of local immunity as well as bacillary elimina- tion were achieved in two thirds of the patients. Further systemic studies are required to establish the power of this agent as an adjunct to MDT in leprosy.

The role of reactive oxygen intermediates in killing of phagocytosed M.leprae by the macrophages of normal healthy individuals and leprosy patients and the causes that were responsible for poor ROI in patients were studied. Experimental materials were macrophages obtained as in vitro culture from the peripheral blood of human individuals. They included healthy normals, bacteriologically positive leprosy patients (B+L), bacteriologically negative long term treated patients (B-L) and tuber- culoid leprosy patients (TT). The fully mature phagocytic macrophages cultured for 7 days were challenged with live or heat killed armadillo derived M.leprae and the H2O2 and O2 released were estimated. The viability of M. leprae phagocytosed by the macrophages was determined by determining the ability of fluorescin diacetate binding to the bacteria while inside the cells. The number of green fluorescin bacterial is the proportion viable among the total acid fast bacteria. In normal macrophages, bacilli with a viability of 56% after phagocytosing was reduced to 21%; whereas in all the other types of macrophages there was no reduction in viability after phagocytosis. The phagocytosed M.leprae were recovered and their ability to grow in the foot pad of mice was determined along with those cells treated with scavengers of superoxide, the superoxide dismutase and that of hydrogen peroxide - catalase. It was clear along with production of O2, viability was lost and when O2 was removed by SOD and H2O2 by by catalase, viability was not reduced. To confirm the role of reactive oxygen intermediates (ROI) in the killing of M. leprae in the cells of normal individuals and inability in the cells of leprosy patients, experiments were carried out in presence scavengers of ROI like superoxide dismutase, catalase, sodium benzoate or thiourea. The data clearly indicated that if ROI production is blocked, the killing of the bacilli inside the phagocytes of normal individuals was blocked. In another set of experiments with phagocytes obtained from leprosy patients, when they were challenged with live M. leprae in in vitro cultures, phagocytosed bacteria were not killed. But if the cells have been exposed to an immunomodulator, it was found that activated super- natant from such DOC exposed cultures were able to modify the macrophage to recognise M. leprae and produce ROI and also kill the M. leprae. This again was blocked by the scavengers. Further experiments were carried out to test whether the early transient viability of M. leprae in the paucibacillary leprosy were also due to failure of the bacilli being killed by the reactive oxygen intermediates. This is important since paucibacillary patients (tuberculoid leprosy type) are endowed with ability to eliminate bacteria through cell mediated immunity, just like normal healthy resi- stant individuals. Yet the immune mechanism in paucibacillary cases is abnormal and this could be due to an unusual type of CMI initiated by a particular set of antigens, unlike the resistant normals. Further, data clearly indicated that macrophages from paucibacillary individuals are unable to recognise M. leprae and produce ROI nor kill M. leprae. The macrophages of lepromatous leprosy patients were found to have unusual features regarding levels of these 3 surface markers as compare to the normal resistant individuals. However, on exposure to the DCC stimulated supernatant, the surface markers were modified to the levels closer to normal macrophages. This could indicate a normalization of the defective macrophages, which then could recognise M. leprae and phagocytose and kill them through ROI. However, such an explanation may not be true for tuberculoid leprosy patients since they do not show the pre-existing surface changes. The only possibility under this condition that DCC as an antigen stimulates the macrophages and T-cells of paucibacillary patients quite differently from that of whole M. leprae to which also the macro- phages and able to respond. In separate studies using antibody coated M. leprae and DCC it was demonstrated that the macrophages of the In separate studies using antibody coated M.leprae and DCC it was demons- trated that the macrophages of the paucibacillary patients appear to have different binding/recognition sites for M.leprae and DCC. This then again indicated a surface disposition in paucibacillary macrophages, perhaps, different from normal resistant individuals. It appears that the basic defect of preexisting membrane perturbations both in the lepromatous and tuberculoid leprosy patients may make the macrophages unable to recognise M.leprae and kill the phagocytosed bacilli through ROI. It was demonstrated that macrophages from normal individuals are able to phagocytose M.leprae and kill them through the production of reactive oxygen intermediates like H2O2, O2 and OH. In leprosy patients such an event does not take place. However, an immunomodulator which has vaccine potential is able to activate the leucocytes of leprosy patients and the culture supernatant of such a process is in turn able to activate the the macrophages to kill M. leprae through reactive oxygen intermediates. The mechanism of activation by the culture supernatant appears to modify the surface properties of the macrophages of the leprosy patients so as to normalise the preexisting defects. Such normalization leads to recogni- tion of M. leprae and their killing through ROI system.

Monoclonal antibodies to delipidified cell components (DCC) of M.leprae were obtained through hybridoma technique and two monoclonals with affinity to 38kDa and 65kDa proteins of DCC have been identified. Other two did not pick up any specific components in the Western blot. One monoclonal helped distinguishing this 65kDa cloned protein as different from classical hsp 65kDa of M.leprae. The monoclonals were not specific but showed reactivity to some other mycobacteria. The mono- clonals can pick up antigens in the sera of lepromatous leprosy patients by ELISA as distinguishable from normal or tuberculoid leprosy sera.

The aims and objective of study the to study the acid fast nocardioform chemoautotrophic bacteria isolated from human and animal tissues infected with leprosy bacilluso. Tissue specimens were obtained form 27 patients attending the department of leprology, School of Tropical Medicine, Calcutta and also from 5 patients attending the leprosy unit of the department of Dermatology, Calcutta Medical college. All these cases showed active clinical disease, and as far as could be known, did not have any antileprosy treatment. Of these 32 cases, biopsies were obtained from 12 patients and slit-skin specimens were collected form the rest. The biopsy homogenates were preserved at -10 centigrade for further work leprosy tissues followed by feshly isolated cultures forn BL/LL cases were subjected to different tests for identity with the leprosy bacillus. These comprised Gram stain, Z-N stain, degree of acid fastness, pyridine extractability of acid fastness, abilities to attack/ utilise collagen, gelatin, liquid paraffin, urea as well as presence of DOPA-oxidase and catalase. It was observed that all the 32 isolates were weakly acid fast, possessed DOPA- sxidase, catalase and gelatinase, and grew luxuriantly in presence of urea or paraffin as the sole source of carbon. Several other studies are now being carried out to find out the similarities of chemoautotrophic nocrodoform (CAN) bacteria with leprosy bacillus. In vitro antibacterial effect of rifampicin, norfloxacin, ofloxacin and other on CAN bacteria was under taken.It was observed that the antibiotics/ chemotharapeutic drugs finally selected are given in tis included inclusion of some new and exclusion of several; 15 CAN isolates could be tested. Ome prospective antileprosy drugs were taken up first for in vitro susceptibity tests on CAN bacteria which could be equivalent of the leprosy bacillus. For this purpose, each drug was incorporated in GMA and also in fluid GM at concentration of 0,1,5,10,20 and 50 mue gram/ ml, taken in McCartney bottles. The bottles were inoculated with small inocula (ca 10000 bacteria), incuibated at 28 centigrade and observed for appearance of growth first after 2 weeks and then twice a week for a period of 3-6 months. Results are given ofloxacin was found to be considrably effective, equalled or closely followed by rioampicin; a minimal antimicrobial activity was noted with norflo norfloxacin. Results of studies wit Na stibogluconate ( a pentavalent antimony compound), urea stibamine, clofazinine, DDS, INH and other quinolones and others are given. This is the first time that an antibiotic sensitivity of in vitro equivalents of leprosy bacillus had been raperted on the basis of conventional antibiotic sensitivity test system.

Surgical nerve deompression (neurolysis) alone or combined with intralesional (perineural) corticosteroid therapy for treatment of nerve involvement in leprosy were evaluated in 61 leprosy patients. Patients studied presented with features of neuritis viz pain, tenderness, palpability and sensory or motor deficit in the ulner nerve territroy of less than six months duration and without atrophy of hand muscles and contracture or deformity of hand. Children below 10 years of age were not included because of difficulty in evaluation of nerve damage. All patients were on anti leprosy drug during the study. Patients were studied in two groups. Group A patients (33) were subjected to neurolysis while Group B patients (28) were subjected to neurolysis plus intralesional (perineural) corticosteroid injection. Nerve function was evaluated prior to neurolysis and at the end 3rd week, 3rd month and 1 year following neurolysis. Patients in Group A, possibly due to perineural corticosteroid. Relief from pain and tenderness was seen in all patients of both groups, but it occured earlier in Group B. Recovery of motor and sensory function also occured earlier in Group B. Recovery was better when the duration of nerve involvement was short (< 3 months>), nerve function deficit less and in patients with paucibacillary leprosy. patients with short segment of nerve involvement with minimal thickening had better recovery. One patient (Group A) with deterioration at one year had fibrous adhesions around the nerve and thickening of nerve was more marked. The results on a small show that in patients on specific drug treatment, progressive nerve damage could be prevented by neurolysis. The beneficial effect of surgery could be accentuated with local corticosteroids at suitable intervals. Combined treatment of neurolysis with perineural corticosteroid injections is recommended when nerve damage is early. However, this requires confirmation on a large sample.

Functional value of nerve decompression in leprosy patients. The nerves of 34 patients of leprosy with nerve involvement ranging from 22 days to 7 years were subjected to nerve decompression to assess its benefit in terms of symptomatic releif and improvement in functions/sensitivity of the afected part ( foot and hand). Due to difficulties in evaluation of nerve damage and improvement, children below 10 years of age were excluded from the study. Twenty five nerves were subjected to nerve decompression alone under local anaesthesia, while the other 25 nerves were treated with intralesional conticosteroid injection intraoperatively, followed by 2 injections at weekly intervals postoperatively. Postoperative recovery of function was assessed at the end of 6 weeks. Sweat prints of the affected part ie palm/foot were taken preoperatively and 6 weeks postoperatively to assess the enhancement of sweat secretion. Neurological involvement was observed more ofter in the tuberculoid leprosy patients (88.23%) and least in the lepromatous patients (5.9%). Loss of sweat secretion and nerve thickening was seen in all patients. Nerve pain and tenderness were seen ( in 84% and 94% patients respectively) in the majority. Improvement in nerve function was observed in most of the nerves subjected to nerve decompression. The nerves subjected to intralesinal steroid injection had better improvement in nerve function. Deterioration or no change in nerve function was observed in nerves with a long duration of nerve involvement and presence of nerve abscesses. Lesser the duration of nerve involvement better was the recovery following either nerve decompression alone or when combined with intralesional steroids. The best results were obtained when nerve involvement was of less than 6 months duration. Marked relief in pain was observed in all nerves subjected to nerve decompression. Enhancement in sweat secretion from the preoperative status was noticed in both the groups which was more marked in patients given steroids. However, the amount of sweat secretion did not reach normal level in either group during the study.

Objective of the project were to study the role of Lipoproteins in nerve damage and foam cell formation in leprosy; to demonstrate that the degenerating and regenerating peripheral nerves in leprosy possess the components of cholesterol transfer mechanism for membrane biogenesis; to trace the cellular association of Apo-E, Apo-A1, Lipoprotein(a) and LDL receptors in various types of leprosy; to study the effect of potential anti leprosy drugs, vaccines or multidrug therapy on these lipoproteins. The development of foam cells is one of the key events in leprosy and the origin of lipids in foam cells has been attributed to LDL and VLDL. Extra cellular lipids in foam cells are thought to be in a dynamic state taking up and degrading cholesterol. The important role of macrophages in the pathogenesis of leprosy to phagocytose bacterial cells has been known but the process of foam cell formation invivo and its possible role in leprosy remains to be elucidated. The circulating levels of Lp(a) a genetic variant of LDL and LDL receptors in lepromatous leprosy are found to be elevated as compared to the Tuberculoids and Controls. These observations implicate the role of LDL receptors and Lp(a) in foam cell formation in leprosy. These receptors have been characterized in terms of their physico-chemical properties to find out if these have undergone any significant alterations due to bacterial infection or is it that onlu the number of binding sites have changed. A double antibody equilibrium RIA specific for Apo-E was standardized in our laboratory and used to estimate the the circulating levels of Apo-E in leprosy patients. These levels were significantly raised and the Apo-E receptors in nerve and skin biopsies showed an increased amout of binding sites in Tuberculoid group as compared to the other groups. The focussing pattern of the delipidated serum by an IEF shows bands in a p1 range from 7-8 in the lepromatous and from 5-7 in the Tuberculoid group.

Objective of the project were to study the role of Lipoproteins in nerve damage and foam cell formation in leprosy; to demonstrate that the degenerating and regenerating peripheral nerves in leprosy possess the components of cholesterol transfer mechanism for membrane biogenesis; to trace the cellular association of Apo-E, Apo-A1, Lipoprotein(a) and LDL receptors in various types of leprosy; to study the effect of potential anti leprosy drugs, vaccines or multidrug therapy on these lipoproteins. The development of foam cells is one of the key events in leprosy and the origin of lipids in foam cells has been attributed to LDL and VLDL. Extra cellular lipids in foam cells are thought to be in a dynamic state taking up and degrading cholesterol. The important role of macrophages in the pathogenesis of leprosy to phagocytose bacterial cells has been known but the process of foam cell formation invivo and its possible role in leprosy remains to be elucidated. The circulating levels of Lp(a) a genetic variant of LDL and LDL receptors in lepromatous leprosy are found to be elevated as compared to the Tuberculoids and Controls. These observations implicate the role of LDL receptors and Lp(a) in foam cell formation in leprosy. These receptors have been characterized in terms of their physico-chemical properties to find out if these have undergone any significant alterations due to bacterial infection or is it that onlu the number of binding sites have changed. A double antibody equilibrium RIA specific for Apo-E was standardized in our laboratory and used to estimate the the circulating levels of Apo-E in leprosy patients. These levels were significantly raised and the Apo-E receptors in nerve and skin biopsies showed an increased amout of binding sites in Tuberculoid group as compared to the other groups. The focussing pattern of the delipidated serum by an IEF shows bands in a p1 range from 7-8 in the lepromatous and from 5-7 in the Tuberculoid group.

The main objective of the study was to implement NLEP activities through Primary Health Care (PHC) staff in areas where MDT was implementd through vertical approach for 5 years or more and study its implications and impact. To achieve this it was planned to access the training requirements of Primary Health Care staff for implementation of activities NLEP, develop a training manual and train PHC staff. To compare and evaluate the NLEP activities when implemented through vertical set up and PHC set up side by side; to find out the impact of NLEP integration on other programmes being implemented through PHC namely family welfare, expanded programme on immunization, maternal and child health (MHC) etc. Continued implementation of conventional NLEP strategies under the existing circumstances of reduced disease burden after implementation of MDT and static status of new case detectin of leprosy is not cost effective. Integration of the NLEP with the PHC system is promising alternative. To study implementation of NLEP activities through PHC staff in areas where MDT was implemented through vertical appraoch for 5 years or more and study its implications and impact. CLTRI Chengalpattu organized a pilot study on integration of NLEP with PHC in two districts of Andhra Pradesh State during 1997-1999. Population covered under the study project was 16,39,307(2,39, 141 study and 14,00,163 control group). The results the study at indicate that prevalence in the study and control area after the implementation of integratin is 2.70 and 1.91 per 10000 respectively. The difference is statistically significant at 95% C.I. level ( p= 0.00001). NCDR is 5.71 and 4.16 per 10000 person years. The difference is statistically not significant (p=0.49). Case holding in stud and control area is 66.3% and 96.9% respectively. This difference is statistically significant at 95% confidence interval (p=0.0001). No difference is observed in the objective performance of the other programmes by the MPHWs in the area. The integration of vertical NLEP activities with horizontal approach of PHC system is feasible, if general health care providers are properly motivated, adequately trained and given the responsibility of implementation. Problems like individual technical incompetency, lack of supervisory and monitoring skills, no referral services, poor record maintenance and other administrative issues were encountered. Such problems can be dealt by providing a specialized component of services which means provision of training facilities, technical supervision and referral services at intermediate and middle level of planning and evaluation.

The major objective of this project were to study the magnitude of the problem of disability before the onset of MDT project and after the completion of the project; to study the deformity rate in the newly detected cases; to study the extent of disability that can be directly related to disease process per se. and that due to inadequate patient care in relation to anaesthesia and muscular weakness. A total of 2547 cases (860 MB, 1687 PB) released from September 1985 to September 1995. Only 2015 subjects (79.1%) could be contacted 612 subjects were MB type. The others had either left the place or died. Out of 2015 cases 113 cses (5.6%) were wrongly diagnosed and 84 (4.2%) cases had not completted MDT. It is the convention not to report grade I disability but in the present study all the the grades I, II and III while evaluating grading. It was observed that grade I disability rate had decreased significantly after MDT in the patients studied. In upper extremities grade I disability rate fell from 3.9% to 2.4% and similary in lower extremities from 3.1% to 2.0%. But it was not due to reversal to normalcy but their patients progressed unfortunately to serve deformities i.e. grade II and III. If only grade II and III were considered a ddefinite increase in disability rate was observed form 6.1% to 7.6% in upper extremities. Siilarly an increase form 4.1% to 5.1% in lower extremities was also obtained. It is worth mentioning that under National Leprosy Eradication Programme the protocol for disability grading enlists and records grades II & III only. The relsuts of present study also highlight disability rate to be 5 to 7 times more common in a multibacillary (MB) cases, if compared wih disability rates in Paucibacillary cases (PB). It is quite evident that deformity has progressed during treatment which could have been reduced if proper care and monthly examination of patient was done in actual practice and in the right ernest. We had observed that the most of the medical officers working under National Leprosy Eradicaition programme examine patients rarely. Similarly para medical wokers (PMWs) are not quite competent enough to recognise the severity of deformity status. Hence, the negligence increases the agony of the patients that is highly in desirable. It means that the drug distributin points are true to their name only where drugs are distributed. Clinical status of disease and complication are rarely and seldomly looked for, hence the peril of the patients continues. Most of the cases with disability had a long duration of disease. Half (51%) of them had disease for a duration of over 5 years before they started MDT, one fourth (24.5%) had the disease for 3 to 5 years. This observation confirms the earlier reports are and it also highlights that if we pick up a patient within 6 months of development of disease that disability could practically disappear from the scene. The reductin in thhe incidence of disabilities can be explained by the efficacy of drugs, tjhe reduction in the duration of the disease, the reductions in incidence of lepra reacitons and by operational factors such as; Improvement in early detection and management of cases. Only visible physical disabilities have been considered here. Temporary physical disabilities before and during treatment, reactions and complication, and the inconvenience of sustaining long duration treatment, should all be considered in order to estimate the impact. Moreover, among all infections and it is difficult to assess the individual social and psychological impact it causes. Estimates based on informations collected from the field through control programmes are often quensioned. It is clear that the sensitivity, specificity and completeness of informations collected should be evaluated. On the other hand, applying rates collected from scientific studies conducted in limited places to a theoretical ' populatino at risk' of leprosyy is also likely to lead to over estimates. Moreover, information on the incidence of disabilities with out intervention and before, during and after treatment is very limited. Hence, it is expected that combinations of methods will give some idea of the complexity of the problem, and will stimulate further work on the collection of reliable data relating to incidence of disabilities.

Aims and the objective of the study waer to evaluate LSR/A15 recombinant protein as predictor/ marker of reactional states in leprosy using conventional ELISA; To characterise and identify B cell epitopes of LSR/A15 that are recognised by the reactional patients; to evaluate LSR/A15 antigen stimulated T cell functions in reactional patients using lymphoproliferative and cytokine with PBMC. Both T and B Cell responses of leprosy patients to LSR/A15 antigen of M.leprae was undertaken on 311 subjects, 20 of whom were healthy contacts in a prospective study with 6 monthly follow up over>3 years. The lymphoproliferative and antibody responses were evaluated with a view to identifying the T cell and B cell epitopes of the LST/A15 as recognised by the leprosy patients during the course of stable as well as reactional disease. Overlapping 10-15 mer peptides as well as end to end peptides with and without C G residues at the termini were used to dissect the epitope recognition by sera and PBMC of leprosy patients. Moreover the antibody response of patients was monitored at 6 monthly intervals. Cytokine release and expression was also examined. In general the epitopes recognised by different ethic populations varied for both T and B cell responses. It was evident that P2 and P3 peptides were recognised by sera from ENL reaction patients but not by those with stable disease. Antibodies to the peptides and LSR decreased during multidrug therapy to negligible levels by 12 to 24 months in the majority of patients. Significantly, 15 subjects continued to show these antibodies and develope ENL reaction by 6 to 12 months. Thus monitoring of antibody titres would have clinical and prognostic significance in ENL reactions. P2 and P3 peptides have predctor role for ENL but not for reversal reactions. IgG subclsaaes also showed differences between stable and reaction patients with predominance of IgG3 and low levels of IgG4 to the peptides in ENL reactions. T cell epitope recognition showed 4 patterns, multi epitope recognition, restricted epitope recognition, single epitope recognition and no epitope recognition. Apparently anergic lepromatous leprosy subjects showed T cell responses with peptides as well as LSR. In general more patitents recognised P2, P3 and LA8 peptides. Cytokine pattern showed the presence of both IFN gamma and IL4 in majority of stable disease patients <40% lepromatous and >50% tuberculoid patients showed only IL4 and IFN gamma respectively. Interestingly during ENL, reactions in lepromatous patients, IL 4 was not detected and INF gamma emerged as the major T cell cytokine. It would appear that dysergulation of IL4 may be the major precipitating factor for ENL reactions.

Aims and objectives with which the scheme was started were to systematically screen clinically and bacteriologically all leprosy affected beggars; to describe the background of those berrars; to examine the family members if any, living with the beggars; to treat those positive with MDT; to develop a continuous maintenance and evaluation system through community organizations for recurrence/ relapse such baggar populations. A pilot study was on 193 beggars in and around Vellore Town, the Head Quarters of Vellore District in Tamil Nadu. Most of them had begged in the same place on most of the occasions. Of 193 beggars, 58 had leprosy with previous history of treatment from leprosy treatment centres. None of them were on any treatment during screening ( All the patients were released from treatment Mono/MDT according to their statement). Out of the 58 leprosy affected persons, 10 (18%) patients showed skin smear positivity with signs of clinical activity they were motivated to attend treatment centres for MDT. The study reinforces the need to screen beggars regularly. Those who turn out to be "positive" should be motivated to continue/ start treatment, guiding them to various centres available. Their close contacts must also be screened frequently, educated to carry out self examination for patches and report to repective centres if they have any suspicion.

The 'P.falciparum' and 'P.vivax' circumsporozerte (CS) proteins con- tains repeat sequences that are mostly conserved in various strains. These repeat sequences are thought to be the target of protective immunity. The repeat sequences of 'P.falciparum' and 'P.vivax' CS proteins viz (NANP) 4 and (GDRADGQPA)2 have been synthesised by solution phase technique. These were characterized by physico - chemical means. Antipeptide antibodies were generated in rabbits using water soluble adjuvant, tuftsin. It has been already established that tuftsina tetra- peptide, helps in antigen presentation to various cells of the immune system especially to macrophages and T cells. The antibody titres are comparable with that generated by Freunds' adjuvant. Hence in vaccine trials, use of tuftsin, as an adjuvant cannot be ignored. Moreever it is permissible to humans, is non-toxic and non-pyrogenic. Further, during the competitive EIA procedure, a minimum structure of three repeats of NANP and two repeats of GDRADGQPA are found to be essential for immunological reactivity. These peptides have been tested as test antigens for detecting anti-CS antibodies to 'P.falciparum' and 'P.vivax' infection in endemic area, where mixed infection is prevalent (i.e. in Ghaziabad district). The results show age dependent increase in antibody levels and the titre is dependent on the parasite density in a particular village. This is the first reported study using defined peptide sequences of P.falciparum' and 'P.vivax' to detect anti CS antibodies, that can discriminate mixed infection. Furthermore, the present work highlights that the Indian strain had more or less conserved sequences within the repeat region.

Detection of material infection (sporozasite stage) in vector mosquitos using monospecific antibodies: The study was carried out in four malaria endemic areas of West Bengal viz. (i) distric Jalpaiguri, (ii) Ajodhya hills of district Purulia, (iii) selected villages in district South 24 Parganas and (iv) selected areas in calcutta city with the objective of collecting information regarding vectors of malaria, their vectorial potential and their relationship to disease. The vecotr species encountered in district Jalpaiguri were Anophelese. An.culicifices, An.minimus and An.fluviatilis. Survey in Ajodhya hills revealed the presence of An.subpictos, An.culicifacies, An annularis, An. maculotus and An.fluviatilis. The anopheline fauna of district South 24-Parganas consisted of 16 species of which six were vectors of malaria viz. An.ennularis, An.fluviatilis, An.subpictus, an.culicifacies, An.fluviatilis and An.sudndaicus. The Culcutta city harboured An.stephersi and very less number of An.subpictus also. ELISA on mosquito-triturates of vector speices of anophelines (An.annularis, An.fluviatislis, An.subpictus and An.maculatus) for the detection of Plasmodium falciparum circum-sporozoite-protein (CSP) using pf-monoclonal and pf-monospeicfic antibodies and thei8r corresponding controls did not reveal the presence of P.falciparum sporozoitas, where as only 3.28% of 3200 An.culicifacies showed the presence of sporozoites An.stephens from Calcutta city also found to harbour P.falciparum sporozoites. Results of blood meal analysis of An.culifacies, An.annularis and An.stephesi revealed that 98.28% An.culicifacies and cent percent An.annularis from catle sheds were boviphilic, where as, 87.5% and 10.22% An.culicifacies collected from human blood respectively. On the other hand 89.79% and 7.4% of An.annularis from human dwellings showed the presence of bovine and human blood respectively. Amongst An.stephensi collected from catle shed 98.4% were positive for bovine blood, 1.03% for human blood, while An.stephensi collectedfrom human dwellings 13.04% had fed on human, 77.17% on bovine blood and 9.78% samples showed the presence of both bovine and human blood in their guts.

The Indian Council of Medical Research (ICMR) initiated an eight centre study in 1985 to improve the coverage and quality of MCH services at the PHC level within the existing norms of the primary health care delivery system. The study was undertaken in the states of Uttar Pradesh, Madhya Pradesh, Rajasthan, Haryana, Maharashtra and Gujarat. A "Comparative MCH Care Pack- age" of intervention was developed adopting the approach of identification and management of high risk factors in the pregnant women and their off- springs. The following strategies were used for implementing the comprehen- sive MCH care package - (i) Re-orientation training of medical and paramedi- cal functionaries to improve their technical/supervisory skills for common high risk factors in pregnant mothers and newborn (ii) Community education to increase their awareness about the high risk factors and utilization of MCH services (iii) Development of better data recording system for monitor- ing and evaluation of MCH services (iv) Developing a feasible referral system for the care of "at risk" mothers and newborn. The collaborating centres had the ICMR's research team representing the disciplines of Obstet- rics and Gynaecology, Paediatrics and Community Medicine as Principal Inves- tigators/Co-investigators as well as the District Health Officer represent- ing the state health authority in the study areas. Periodic meetings at regular intervals were held to ensure proper coopeation and coordinati between the ICMR's research team and the District/State Health Authorities. The Primary Health Centres (PHCs) were identified for the study according to the selection criteria laid down by the ICMR such as (i) No other active intervention programme should be ongoing in the study area and (ii) The infant mortality rate (IMR) should be similar to that of the respective state. The situation analysis which lasted for six months revealed that the population covered by each of the eight PHCs was variable, ranging between 80,000 to 1,69,000 except in the state of Maharashtra where the Government of India's norm of 30,000 population per PHC had already been implemented. Except for the equipment and vaccines under the Universal Immunisation Programme (UIP), the availability of otehr facilities in terms of physical space, other general supplies and equipment including the drugs were poor at the PHC level. In general, the availability of manpower was satisfactory as per norms. The provision of MCH services was limited to providing the Iron- Folic Acid (IFA) tablets and Tetanus Toxoid (TT) immunization. There was no concept of detection of high risk factors by Auxiliary Nurse Midwives (ANMs) in pregnant women/newborns or for their better care by having a suitable referral system. The quality of care was poor during antenatal, intranatal (including the use of safe delivery kits) and postnatal periods. The data recording was poor and reflected gross underreporting of births, deaths a other vital statistics. The gaps which were identified during the situation analysis were bridged in terms of strengthening infrastructural facilties with the help of state health authorities and also by the ICMR's research team as required during the preparatory phase. The other activities during the preparatory phase included the preparation of training manuals, health educaitonal materials and development of systematic data recording forms such as MCH cards and registers. A baseline survey was also done during the preparatory phase to obtain the household information and MCH/FP practice in the study area as well as to determine the denominators of pregnancy and vital rates. The otehr important activities during the preparatory phase included the initia- tion of re-orientation training programmes for health functionaries, commu- nity education activities, improving the record forms and data feedback system and establishing a referral system. The package of interventions for the comprehensive MCH care utilizing the high-risk approach strategy was initially implemented at the one third area of the PHCs in the 30,000 popu- lation for a period of two and half years (30 months). Thereafter, these interventions were expanded to the remainder of the two thirds of the PHC study area. As a result of above interventions, there was a significant improvement the quality and coverage of MCH services. For example, there was a progres- sive increase of registration of pregnant women during the intervention period - 2282 women in 1988, 5202 women in 1989 and 5423 women in 1990. The majority of pregnant women registered, about 77 percent, were between 19-28 years of age. The average incidence of pregnancies in women having less than 18 years of age was 6 percent, except a higher incidence of 14 percent observed in the PHC at Maharashtra. About 20 per cent of registered women were primiparous and 17 percent were para 4 and above. Apart from the progressive increase in the registration during the intervention phase, there was an encouraging trend of increase in early registration of pregnant women who were less than 19 weeks of pregnancy. Besides increase in number of home visits by the health functionaries, the coverage of pregnant women also showed an increase for IFA tablets distribution from 53 percent to 71 percent and for TT immunization from 61 percent to 82 percent. While majority of 73 percent of deliveries were conducted at home, it was heartening to note that there was an overall increase in the use of Trained Birth Attendants (TBTAs) by the pregnant women during the intervention period. The birth weight recording which was non-existent before, showed a significant increase of its recording by spring balance in 69 percent of newborns. A decreasing trend of the babies being born of low birth weighi less than 2499 gms. was also observed during this period. Apart from an icnrease in the postnatal visits from 68 percent to 75 percent by the health functionaries, visits by them within 24 hours also showed an increase from 39 percent to 45 percent. Earlier, there was no practice of identification of commonly known risk factors in rpegnant women and newborns by the health functionaries nor any referral system existed for their better care. As a result of training progrmames,,, the health workers reported an overall incidence of one or the other of high risk factors in pregnant women to around 35 percent during the intervention phase. The prevalence of commonly known high risk factors was greater in women who were either in the ages of less than 18 years of over 30 years or who had an interpregnancy interval of less than 18 months or who had a previous bad obstetric history. Amongst the pregnant women,,, the incidence of multi-gravida (>5) was 15 percent and about 9 percent of the women had previous bad obstetric history. Amongst the newborns, the risk factor of low birth weight i.e., less than 2499 gms. birth weight was present in 12 percetn and early delivery at less than 37 weeks of gestation was seen in 6 percent. It was observed that amongst those pregnant women in whom one or the other risk factors were present, their pregnancy outcome was adversely affected as indicated by the higher incidence of low birth weig babies. It was encouraging to note that no difference in the provision of various types of MCH services was rpacticed by the health functionaries during the antenatal period to the pregnant women who had either no risk factor or who had one or more of the high risk factors. However, as a result of training progrmames, these health workers provided greater concern and intensity of care to the pregnant women in whom the presence of one or more of the high risk factors were identified. For example, the pregnant women who had the high risk factors were registered earlier i.e., at less than 20 weeks gesta- tion. These women had more hom visits by the health functionaries during the antenatal period and a significantly higher percentage of them were seen by the medical doctors. A greater awareness was observed amongst the health functionaries for the referrals since 18 percent of women having risk fac- tors were referred by them for better care as comapred to 7 percent of referrals for pregnant women having no risk factor. As a result of communi- ty education programmes, the community awareness of the utilisation of referral services for high risk pregnant women was greater. For example, 53 percent of high risk women had utilized the referral services as compared to 45 percent of women who had no risk factor. An important finding was that the greater weightage for the utilization of referral services was given the pregnant women to the age factor. For example, the pregnant women having the ages of less than 18 years or over 30 years had availed 65 per- cent and 50 percent of referrals respectively. Another important stimulus for the utilisation of referrals was the presence of clinically apparent symptoms of oedema, eclampsia or convulsions since 67 percent of the preg- nant women having one of these symptoms had utilized the referrals. It was important to note that 75 percent of referrals were made to the subcentre (SC) level. Furthermore, the outcome of pregnancy was better amongst the risk women who had utilzied the referral services, since incidence of low birth weight babies was 26 percent as compared to 35 percent in women who did not avail the referrals. In summary, the comprehensive MCH care package of interventions utilizing the high risk approach resulted in a significant improvement in the coverage and uqality of MCH care for pregnant women and newborns at the PHC level within the norms of existing primary health care delviery system in the country. The data also indicates that while there was a general improvement in the MCH services, better care and referral services could also be provid- ed to the rpegnant women having onr or the other commonly known high risk factors. Furthermore, there was an increase in utilization of MCH services by the pregnant women. The experience gained and the lessons learnt fr the present study can be suitably applied to the Child Survival and Safe Motherhood (CSSM) initiative being udnertaken in India and other devleoping countries.

The present study was conducted with the notion that elevated free oxygen radical productiom can induce the pregnancy induced hypertension due to damaging effect on endothelial cells found in PIH. The aim of this study was elevate the blood levels of oxidative stress assessed by measuring malondialdehyde, antioxidant enzymes and antioxidant factors in women with PIH comparing these levels to healthy pregnant women and non pregnant healthy women. Total 190 women were recruited, 10 were non pregnant healthy women, 56 were normal pregnant women, 124 women with pregnancy induced hypertension ( PIH) which were again classified into two categories; 77 women with sever PIH ( blood pressure = 171/114) and 47 with mild PIH (blood pressure = 138/100). Average period of gestation were about 36 weeks. Oxadative stress was assessed by measuring MDA levels in blood, were significantly higher in severe PIH women. Antioxidant enzymes viz; superoxide dismutase and glutathione peroxidase activity in blood were significantly decreased in pregnancy induced hypertensive women than normal pregnent women as compared to non pregnant women indicaiting an adaptational phenomenon to combat oxidative stress associated with pregnancy. However, catalase activity in blood was not significantly altered among PIH groups, while it was significantly higher in non pregnant women as compared to PIH women. This shows that the neutralization of free radicals by calalase in pregnancy is hampered. These findings suggested that pregnancy itself may have deleterious effect on catalase activity. Antioxidant factors like reduced glutathione levels were significantly decreased in normal pregnant women, women with MPIH as well as SPIH and as compared to non pregnant women. In contrast, vitamin E levels which act as antioxidant were significantly elevated in pregnancy induced hypertension as compared to that of normotensive pregnant women. Vitamin C leves were higher in pregnant women compared to non pregnent women and further increased significabntly in women with mild PIH. The increased vitamin E in SPIH and vitamin C in MPIH could be associated with increased utilization of reduced glutathione. A significant positive correlation was observed bteween MDA and catalase activity in severe PIH and a negative correlation with GSH as well as vitamin C with respect to increased blood pressure. It appears that this imbalance between oxidant antioxidant is the effect of disease and not the dausative factor and hence unnecessary supplementation of exogenous antioxidants in pregnancy may actually be not helpful for combating PIH, as any excess of antioxidant in pregnancy may it self affect the fetus at time.

Study was carried out to evaluate the potential anticancer activity of Crocus sativus and Saraca asoca flowers by in vitro cytotoxic studies using various cell lines like DLA, S-180, Hela, Vero etc. Both the extracts inhi- bited the growth of ascites tumours and solid tumours experimentally induced in mice. Oral administration of the extracts reduced the size of 20-methyl cholanthrene induced fibrosarcomas in mice while the same extract prevented growth of papillomas induced by 7,12 dimethyl benz (a) anthrecene in mice, besides delaying the onset of papilloma formation significantly. The extract of saffron was found to enhance the therapeutic efficacy of cyclo- phosphamide treated mice when given in combination. Many of the side effects of cyclophosphamide treatment like leucopenia, fall in haemoglobin levels and increase in enzyme levels were effectively modulated by combined admini- stration of saffron extract. Studies showed the pharmacological properties of the extracts as anti- tumour chemopreventive and a chemoprotective agent.

Filaricidal potential of some plant substance. Rhizome extract from Zingiber officinale, essential oils from the leaves of Anthocephalus morindaefo and Xanthium strumarium and solamargine from Solanum viarum were tested in vivo on Setaria cervi transplanted in albino rats, Acanthocheilonema vitae in multimamate rate and Dirofilaria immitis in parish dogs for their antifilarial activity. DEC was used as a reference standard. None of the plant substances showed any toxic effect when given by oral route (500 mg/kg/d for 10 days for solamargine, 400 mg/kg/d for 10 days for A.morindaefo was not toxic by subcutaneous route at 60 mg/kg/d for 10 days. Z.officinale at 200mg/kg/d (total 8 g/kg in 40 days), A.morindaefo at 200 mg/kg/d (total 9 g/kg in 45 days), and at 45 mg/kg/d by subcutaneous route (total 675 mg/k in 15 days), X.strumarium at 200mg/kg/d (total 830 mg/kg in 5days), DEC at 200 mg/kg/d (total 1 g/kg/d in 5 days) and 111 combination of solamargine and rhizome extract of solarum varium at 166 mg/kg/d ( total 830 mg/kg in 5 days) showed varying degrees of microfilaricidal activity. The combination drug (Solamargine + rhizome extract) at a relatively low dose produced a sustained antifilarial effect and was, therefore, most promising. A.morindaefo also showed macrofilaricidal action eliminating 70-80% adult worms. The subeutaneous route was ferrnd to be more effective compared to orale reite.

Sulphur containing compounds viz Diallyl disulphide (DADS), Diallyl sulphide (DAS), Allyl methyl sulphide (AMS) and S- allyl cysteine sulphoxide (SACS) present in garlic were studied for their immunomodulatory, anticancer and radioprotective acitvities in Babb/c mice. Intrapentonecell administration of sulphur compounds was shown to stiumulate the haemopoetic system as seen from the incrreased total count bonemmarrow leneocyte cellularity and alpha esterase positive cells. Treatment with sulhur compounds antibody increased the curculating titre and number of antibody producing cells indicating the stimulation of humoral immune response. The cell mediated immune responses in normal as well as tumour bearing animals were alsoenhanced by the administration of sulphur compounds enhanced by the administration of sulphur compounds as seen from the increased NK cell and antibody dependent cellular cytotoxic activities. Sulphur compounds enhanced the production of cytokines such as IL-2, IFN-8 and GM-CSF in normal animals. The lowered levels of these cytokines in the cyclophosphamide treated animals were brought back by the treatment with these compounds. Sulphur compounds exhibited tumour reducing activity. Administration of these compounds could significantly inhibit the solid tumour development induced by DLA cells in mice. Similarly these was a significant inibition of sarcoma development induced by 20 methyl cholanthrene. Further, the sulphur compounds showed a significant radio protective effect. DADS. Protected the haemopoietic system of radition exposed animals as seen from the enhanced bone marrow cellularity and alpha esterase positive cells. Sulphur compounds also could significantly reduce the chromosomal damage induced by radiation exposure. The results are indicative of the usefulness of sulfur containing compounds from garlic in the prevention and management of cancer.

Evaluation of the effectiveness of brief in patient family intervention vs. out patient intervention for mentally retarded children. A prospective study was carried out on mentally retarded childrento evaluate the effectiveness of brief in patient and out patient family focussed interventions (developed in house) in terms of their impact on the childs development and behaviour, and family adaptation (preceived stress and coping). The components of the interventin included medical measures, family oreientation, general parenting measures, and parent training. Outcome variables studied included child variables (intellectual/ adaptive functioning and presence and serverity of problem behavviour) and family variables (self report and observer rated stress in families and level of family adaptation). A total of 157 mentally retarded children of either sex (75 in the in patient group and 82 in the out patient group) from low socio economic group (both rural and urban) and with moderate to profound degree of mental retardatin were recruited for the study. Children who had already undergone major interrvention, those with sever hearing or vision impairment which was more disabling than the mental retardation and children with progressive neurologic disorders were excluded from the study. Thirty three (44%) children/ families is the inpatient and 47(57%) in the outpatient group could commplete one year of follow up. Improvements or gains were noted on all outcome measures to a varying extent in both the groups. The gain in the child adaptive functioning was more pronouned in the inpatient group (10 points) as compared to the outpatient group (4 points). Problem behaviour came down by 55-60%, family stress reduced by 27-35% and familyadaptation was improved by 30% in both the groups. Maximum gains occurred during the first 3-6 months of follow up and subsequently remained stable over the remainder of the follow up period. There were no major effects of age, sex, and residence of outcome. It is therefore concluded that it is possible to formulate and test effective family focussed interventions in mental retardation which are suited for Indian conditions and such improvenments in both children and their families.

The study was carried out to find out incidence of Aspergillus species in the sputum of normal healthy persons. The incidence of antibodies in normal healthy persons was also determined. The incidence of aspergillosis in bronchial asthmatics and in allergic pulmonary infections was studied. Estimation of immunoglobulin levels including IgE in Pulmonary aspergillosis and in allergic pulmonary infections was done. The sputum samples were collected from patients admitted to KMCH, Manipal with chronic respiratory tract infections like Pulmonary tuberculosis, bronchial asthma, chronic obstructive pulmonary disease (COPD) suspected allergic bronchopulmonary aspergillosis (ABPA), bronchiectasis and reactive airways disease. Sputum samples collected on 4-6 consecutive early morning samples were examined by direct microscopy with KOH mounts, Gram's and Z.N. stain methods. The specimen were subjected for concentration method by using N-acetyl cystine hydrochloride and after concentration the specimens were inoculated into SGA, LJ media, blood agar (aerobic and anaerobic) RCM, BHI agar. The serum samples from clinical cases and normal healty persons were sub- jected to serological investigations. Antigen for serological tests was pre- pared; Aspergillus Hyper Immune serum in rabbit were raised; Gel diffusion tests were done; Countercurrent immunoelectrophoresis was done; Immunoglobulin estimation of IgG, IgA and IgM was done by Radial Immunodiffusion using Tripartigen plates. Estimation of IgE as per the methods Total IgE by RIST and specific IgE by RAST technique. Elisa test was carried out; Skin tests-Antigen for skin test was procured from V.P. Chest Institute Delhi and the tests done on Volar aspect of skin of fore- arm. I.D. The reading of skin tests was done as per the instruction provided with antigens. Out of 825 cases studied, 127 cases (15.3%) grown Aspergillus in sputum repeatedly. A fumigatus in 92 cases (11.15%) A. niger in 27 cases (3.2%) and A. flavus in 8 cases (9%). A total of 160 (19.3%) cases showed superpositi- vity by CIEP. CIEP is more sensitive than gel diffusion technique. Out of 406 normal healthy persons, none of the sputum samples, yielded Aspergillus repeatedly. Gel diffusion and CIEP were positive in 5 cases (1.2%). ELISA was positive in 3 cases (1.4%) out of 202 cases. Out of 151 cases of immunoglobulin estimation, IgG was raised in 115 cases (76%). IgM in 110 cases (72.8%) and IgA in 84 cases (55.6%). Out of 130 cases of IgE estima- tion, 106 cases (81.5%) positive of PRIST and 53 cases (40.7%) positive for RAST. None of 50 cases of normal healthy persons showed any positive PRIST or RAST. Out of 213 cases of Tested for ELISA, 145 (68%) were positive for Elisa. Only one out of 100 normal healthy persons was ELISA positive. Out of 117 cases skin tests, 46 cases (39.3%) were positive for Aspergillus antigens. Seropositivity was more common above the age of 50 years, in males than females.

A total of 669 patients with chronic lung diseases and malignancies with suspected fungal lung infections were investigated. The patients with haematological malignancies, bronchogenic carcinoma, pulmonary tuberculosis pneumonia, lung abscess with pneumothorax, chronic bronchitis with bronchiectasis, aspergilous lung diseases and allergy, comprising bronchial asthma, allergic bronchopulmonary aspergillosis (ABPA), aspergilloma and miscellancous cases with chronic obstructive airways disease, secondaries in lungs, pulmonary eosinophilia, cavitary disease of lungs, diabetes mellitus, chronic lung infiltrates and P.U.O were selected for the study. Specimens for culture collected were sputum and throat swab on three consecutive days alongwith bronchial aspirate, tracheal secretions and pleural fluid whenever needed. Serological tests performed for demonstra- tion of antibody were (i) Immunodiffusion test for antibody against Candida, Aspergillus and Histoplasma (ii) Counter Current Immunoelectrophoresis for Candida and Aspergillus (iii) whole cell agglutination for Candida and Cryptococcus (iv) slide latex agglutination for cryptococcal antigen detec- tion (v) Enzygnost IgE monoclonal kit test (Behring) for total IgE estima- tion (vi) Phadebas kits and Phadozyme (R) kits and avidin-biotin ELISA for specific IgE against Aspergillus fumigatus and (vii) skin test against aspergillin in patients with bronchial asthma, ABPA and aspergilloma cases for demonstration of type I, type III and type IV hypersensitivity. Out of 51 cases with haematological malignancies, 19 patients were found to be positive for significant growth of Candida albicans, one for C.tropicalis and 3 for Aspergillus flavus Serology was positive and correla- ted in 13 patients with significant growth of Candida and in all patients for Aspergillus. Of the 6 cases with bronchogenic carcinoma, 2 were positive with significant growth of C.tropicalis and one for C.albicans. Serology was also positive in two cases. Of the 110 cases with pulmonary tuberculo- sis 53 patients showed significant growth C.albicans (5), C.tropicalis (1) C.krusei (1) C. guilliermondii (1) A flavus (1) and A.fumigatus (1) Serology performed in 26 patients showed higher titres of Candida agglutinins in 11 out of 12 cases showing significant culture. Strong precipitin bands were seen in one patient with significant growth of A.fumigatus on culture. Significant culture of C.albicans (11 patients). C.tropicalis (7 patients), C.parapsilosis (1 case) and A.flavus (1 case) were found amongst 85 cases of pneumonia. Serology done in 25 patients confirmed significant growth of Candida in 14 patients and A.flavus in 1 patient. Out of 58 cases with chronic bronchitis bronchiectasis, lung abscess/pneumothorax, significant growth or Candida was seen in 25 cases and Aspegillus. flavus in 2. Serology performed in 14 cases, confirmed significant growth of Candida in 4 and Aspergillus in 2 patients. Amongst 220 cases with miscellaneous disorders, significant cultures of candida were seen in 102 patients, A. flavus in 2, A. fumigatus in 2 and C.neoformans in 1 patient. Serological investigation was available in 60 patients. It confirmed significant cultures of candida in 22 patients, A.fumigatus and in one patient Cryptococcosis. Of the 19 bronchial Asthma cases all were negative for Aspergillus on culture and ID test. Type I hypersensitivity was positive in 7 patients who also showed raised total IgE in serum. Specific IgE was best detected by RIA and and ELISA test. Twenty three cases of ABPA showed positivity by cultures for Aspergillus Spp. in 9 patients (A. fumigatus-4, A. flavus-2, A.niger-1, A.fumigatus+A. flavus+A.niger-2). Type I hypersensitivity was observed in 18 and type III in 3. Total IgE was raised in 17 out of 18 patients (94.4%), specific IgE was observed by RIA in 6(33.3%), ELISA in 9(50%) and AB-ELISA in 7(50.3%) patients. Amongst 6 patients with Aspergilloma, Aspergillus culture was positive in all (A.fumigatus-3 and A.species-3) Serology by ID was positive in all patients. Type I hypersensitivity and raised total IgE was observed in 3 patients although specific IgE was negative by all tests. Significant fungal isolation was noticed in 38% patients with testing of single sample, as compared with 61.5% with 2-4 specimens and 77.3% with 5 or more samples. Thus 5 or more samples of consecutive days helped in detecting more cases of respiratory fungal infections.

A total of 807 cases were studied and 259 characterised as nonspecific vaginotos if the vaginal discharge fulfilled at last three of four criteria suggested by Memsel et al. Swabs were processed for isolation of various aerobes and anaerobes. 53.7% of NSV cases revealed that G. vaginalis is the most common organism (p>.001) associated with NSV either as a sole pathogen or in conjuction with anaerobes. Bacteroides and peptostreptococci species are the common anaerobes associated with G. vaginalis. 68.3% of NSV cases revealed plymicrobial etiology. Columbia agar base with 5% human blood nalidixic acid and gentamycin support good growth of G. vaginalis. Colonisation of the organisms in the vagina during different gestational stages of pregnancy had no ill effect of its outcome i.e. is not a maternal hazard. Of the 70 infertility cases 24% were colonised with G. vaginalis but the number is small to draw a definite conclusion. Biotyping of G. vaginalis as suggested by piot et al is an easily adoptable procedure and our findings of biotype 1 and 5 in greater frequency correlated well with other workers. It also suggests that there is no significant difference in the geographical distribution of the different biotypes. Adhesive properties of some of the isolate were studied with the haemagglutination reaction of G. vaginalis isolates from clue cell positive and negative cases with human, sheep and chick RBCs. It showed on correla- tion with the clue cell phenomenon. Hence clue cell formation may be due to other factors. Tissue culture study may throw light on this and the pathogenesis.

This study was undertaken to determine the prevalence of Non-specific vaginitis (NSV) and some associated microorganisms in women attending the GOPD Gyane out Patients's Department and Family Welfare Clinic of LNJPN Hospital, New Delhi. Eight hundred and two (802) women were included in the study, 544 as symptomatic cases of vaginitis and 258 asymptomatic controls. Trichomonas vaginitis was found in 5.5 percent and Candidal vaginitis in 6.9 percent of the cases. NSV was found in 50 percent of the symptomatic women, but also among 18.9 percent of asymptomatic women enrolled initially as controls. M.hominis was found to be significantly associated with NSV among CuT users (P<0.01) while G.vaginalis was significantly associated with NSV among non-CuT users (P<0.001). NSV and culture positivity for G.vaginalis were associated significantly with presence of excessive vagninal discharge, itching and malodour (P<0.001). A positive amine test and predominance of gram negative or gram variable coccobacilli in vaginal fluid gram's smear were found to correlate very well with NSV and G.vaginalis infection. Among the epidemiological factors studied, a duration of less than or equal to 6 years of married life was significantly associated with presence of G.vaginalis (P<0.01). NSV was more common among women with a parity of more than 2, in their postovulatory stage and were more than 22 years old (P<0.01). G.vaginalis isolates were biotyped on the basis of hippurate hydrolysis, production of lipase and beta-galactosidase (ONPG). The most prevalent biotypes were 5,2 and 1. However, no correlation would be made between any particular biotypes and NSV. Cervical cytology was possible in 441 subjects only out of the total study population and inflammation was present in 350 subjects. A statistically significant association was found between inflammation and presence of G.vaginalis o nn culture and NSV. Since the initial study did not reveal any correlation between clinical disease and epidemiological characterization of G.vaginalis by biotyping, further characterization of G.vaginalis isolates was carried out by antigenic analysis of whole cell proteins by sodium dodecyl-polyacrylamide gel electrophoresis (SDS-PAGE). The antigenic profile of G.vaginalis was compared with that of other vaginal isolates that may resemble G.vaginalis on the basis of morphology, culture and biochemical characteristics. Any variation in the antigenic profiles amongst G.vaginalis strains in respect of clinical source and biotype was also looked for. Majority of the strains showed presence of 25-30 protein bands (approximate molecular weight 17kD-20 kD) with an average of 28 bands, with prominent bands which migrated to a distance corresponding to a molecular mass of 90,74,53,51,49 and 41 kD. Visual inspection of gels showed similar patterns in all the strains of G.vaginalis and the similarity calculated by dice coefficient ranged from 73.1 - 100% (average 88.9%). G.vaginalis strains recovered from different clinical groups (NSV, Non NSV and Healthy) or belonging to different biotypes did not differ with regard to their SDS-PAGE antigenic profile or pattern. The antigenic profile of vaginal diphtheroids and unclassified coryneforms organism (U.C.O.) was quite distinct from that of G.vaginalis, the vaginal diphtheroids showed presence of 12-14 protein bands with a wide and prominent band at 140 kD and majority of U.C.O.'s showed presence of 2-3 faint protein bands only. One vaginal isolate that was initially identified as an U.C.O., was subsequently identified as G.vaginalis on the basis of its antigenic similarity to the other G.vaginalis isolates. In an attempt to develop a serotyping scheme, antisera to six vaginal isolates of G.vaginalis were raised in rabbits, however, only one of the antisera showed presence of precipitating antibodies against its immunising strain.

Study was carried out to isolate Candida from various human infections and to evolve a scheme for species identification based on sensitivity to dyes and chemicals and compare it to conventional techniques. A total of 1031 specimens were collected from various types of infections and Candida species was isolated from 433/1031 speciments. All the 433 strains were first speciated by conventional methods and then by disc diffusion tests. The chemicals/dyes used in the disc diffusion test were Brilliant green, Janus green Cyclohexamide, Fast green, Rhodamine 6G, ethidium bromide, Triphenyl tetrazolium chloride, and sensitivities were coded in that order. Majority of strains of C.albicans had the code 1200560, C.tropicalis 1230567 C.krusei 1000060, C.guillermondi 1230560, C.parapsilosis 1000507, C.stellatoideae 1000500 and C.pseudotropicalis 1000567. In the case of C.stellatoideae and C.pseudotropicalis all strains of each species gave a single code but the number of strains tested was small. All other species gave more than one profile with majority of strains being represented by one code ie., majority of strains of C.albicans had the code 1200560 but some strains had profiles which deviated slightly from typical strains. The only code which represented more than one species was 1200567 which was seen in 15% of the strains of C. albicans and 4% strains of C. krusei. This was the only instance of profile overlap and these strains could be readily differentiated by other properties such as pellicle formation in liquid media by C. krusei. The disc diffusion test appears to be reliable method of speciating candida isolates from clinical specimens, because it is simple, inexpensive and gives reliable results in 24 hours.

A total of 10 hospitalised patients from May 89 to Sept. 90 with bacteriologically proven meningococcal meningitis were enrolled in the study from neighbouring states of Haryana, Himachal Pradesh, Punjab, Chandigarh and in Panchkula (near Chandigarh). Out of these, 2 patients expired. For antigen detection, latex agglutination and CIEP was performed and was positive in 9 out of 10 patients on CSF, blood and urine. A total of 40 contacts of these patients were also examined for carriage of meningococci. It was found that only 2 out of 40 contacts (not related) were positive for meningococci. A total of 70 sera from subjects vaccinated already with meningo A+C vaccine Biomerex were collected. Antibody response against N.meningitidis A in 25 hospitalized cases of meningococcal disease, 71 persons in close contact with the above cases and 95 (50 individuals vaccinated with A+C vaccine and 45 "old" vaccinated) were studied. During the acute stage of the disease, haemagglutination antibodies could be detected in 32% and IgG antibodies by ELISA in 16% of samples. During the convalescent period both IHA and ELISA showed significant rise of antibody in 80% of the samples. In the 31 contacts of the patients antibody was detected in 58% and 38% by IHA & ELISA respectively. In the vaccinees, specific Haemagglutination antibody was detected by IHA in 56%, 82%, 78%, 74% of the pre, one month, three months and six months post vaccination respectively. ELISA test could demonstrate antibodies in 2%, 80%, 78 & 66% of the four respective groups. On the basis of the study it is concluded that the active serological response to meningococcus group A does occur in patients, contacts and following vaccination. Individuals should be followed for longer periods in this part of the country for the efficacy of the vaccine. Meningococcal vaccines manufactured by other concerns should also be studied & checked for the serological response they evoke in set up.

Outer membrane proteins were prepared from Bacteroides fragilis, growing the organisms in brain heart infusion broth supplemented with growth promoting factors. SDS-PAGE of antigens prepared from organisms cultured have been similar polypeptide patterns in the two strains tested. Seventeen to twenty polypeptide bands in the range of 14 to 116 kD were seen in all outer membrane preparations. Polypeptide bands corresponding to 31.40kD, and 58kD were present in antigen prepared from organisms grown in culture conditions like brain heart infusion broth as such, when supplemented with cholesterol, cystine and methionine separately and all together. 21.5 kD band was present in antigen prepared from organism grown in culture conditions like BHI supplemented with cystine and cholesterol i.e. 7th and methionine and cholesterol. Antigens prepared from culture were used in immunoblotting. Various immunodominent bands were detected i.e. 116 kD, 66 kD and 42 kD in antigen preparations when sera from mice infected with B.fragilis were tested. These immunodominant antigens will be further evaluated as an immunogen for a possible candidate vaccine against 'B.fragilis' infections.

The study was carried out with the objective of screening the preva- lence of chlamydia trachomatis infection in genital tracts of symptomatic and asymptomatic women. Symptomatic group constituted 125 women attending S.T.D. out patients dept. and 375 women attending gynaecology out patient dept(OPD) asymptoma- tic group consisted 250 each of apparently healthy women attending antenatal O.P.D. and family planning O.P.D. Endocervical specimens were collected from these women and screened for C. trachomatis by enzyme immuno assay. The results revealed that C. trachomatis is prevalent in symptomatic women attending S.T.D. and Gynaecology O.P.D. The O.P.D. as compared to the women attending gynaecology O.P.D. The prevalance of C.trachomatis is low in asymptomatic women. However group of women with Cu 'T' in situ showed a high incidence of chlamydial infection. C.trachomatis infection was observed to be more in young women (<20 years). Data also revealed that C.Trachomatis infection is associated more with pelvic inflammatory disease than with any other clinical condition.

The study was carried out on 126 multiple drug resistant strains of salmonella typhimurium obtained from the National salmonella Phage Typing Centre, New Delhi (collected from different geographical regions of India), to define potential virulence factors viz. enterotoxin production haemagglutination properties, and adhesive and invasive traits. The plasmid profiles, phage types and antimicrobial resistance patterns of S. typhi- murium strains and the association between plasmids and some of the virulence factors were also studied. All the strains produced mannose resistant haemagglutinin and bound to HeLa cells to varying degrees. These strains also potentially invaded HeLa cell monolayers. Plasmids did not influence the adhesive properties of S. typhimurium as revealed by triparental mating studies. Plasmid positive wild type strains and its cured clones (without plasmid) more or less equally adhered to HeLa cells. Mannose resistant haemagglutinin (MRHA-B) encoded on non-autotransmissible plasmids. Hence it is possible that adhesive and invasive traits in S. typhimurium may be partially controlled by chromosomal and plasmid genes. Enterotoxigenicity of S. typhimurium was tested in two biological assay system viz. rabbit ileal loop test and skin permeability test. Enterotoxigenicity was observed in 30.5 per cent strains with 17.8 per cen showing high enterotoxigenic activity in culture supernatant. The most common R-patterns were ACKTSSxTp followed by ACKSSxTp and ACTSxTp. Prevalent phage types were 99,36 and 32. Most strains (117) were untypable. Plasmids were of different molecular weights viz small molecular weights: 5.4-1.8 Md, large molecular weight: 114.3-73.5 Md and intermediate molecular weight: 35.8-22 Md Different plasmid patterns corresponded to a specific R-pattern while the converse was not seen. Phage untypable strains could be grouped into different plasmid profiles. Plasmid pattern determi- nation could be used as an epidemiological tool in a large geographic region as the plasmid profile of a given clone was zone specific.

Studies were carried out to compare the effect of different modes of administration of human tetanus immunoglobulins in tetanus (intramuscular, intrathecal and combined), decrease the morbidity and mortality in patients suffering from tetanus, shorten the hospital stay of the tetanus patients and assess the efficacy of human tetanus immunoglobulins (HTIG) in treatment of tetanus patients. 90 patients aged 15-65 years admitted to the adult tetanus ward of Nehru Hospital, Post Graduate Institute of Medical Education and Research, Chandigarh, between December 1990 and December, 1992 were divided into three therapeutic grades (mild -I moderate - II, severe - III) of 30 patients each. The three grades were further divided into three groups (I, II & III) depending upon route of administration of human tetanus immunoglobulin. The sub group I received intramuscular 30 IU kg-1 body weight of human tetanus immunoglobulin, group II received 500 IU of human tetanus immunoglobulin intrathecally and group III received 30 IU kg-1 body weight and 500 IU intrathecally of human tetanus immunoglobulin. The other treatment intramuscularly included sedatives, non depolariser muscle relaxants and artificial respiration. survival of patients in the present series was 81.11%. Patients who received combined intramuscular and intrathecal human tetanus immunoglobulins showed statistically significant higher survival (100%) as compared to either mode of administration alone. Hospital stay of these patients was relatively less as compared to either mode alone. Patients with incubation period of 7 days and more and patients administered those with period of onset more than 24 hours had better survival. Most common cause of death was severe chest and respiratory tract infection followed by peripheral circulatory failure and autonomic imbalance. Maximum mortality was observed during first 8 days after admission to hospital (64.7%). Incidence of tetanus was more in rural population (93.33%) and in patients who sustained trauma (58.88%).

Nitrogen laser showed no obserable effect on the colony morphology, viable count and bio-chemical characters of Esch. coli. There was an observable decrease in the number of colony forming units after Nitrogen-dye laser exposure. Carbohydrates were not fermented by the organism after Nitrogen-dye laser exposure for 70 mins. Fermentation of Dulcitol was negative after Nitrogen-dye laser exposure for 40 mins. Indole production was affected after the exposure for 60 mins. Amino acid decarboxylation; Orni thine decarboxylation became negative after the laser exposure and Arginine decarboxylation became positive after exposure. He-Neon laser showed no observable effect on biological, biochemical characters studied. There was no effect on the antibiotic susceptibility pattern of Esch. coli after the exposure of Nitrogen-dye and Helium-neon laser, whereas the organism, which was sensitive to sulphadiazine became resistant after Nitrogen laser exposure. When compared to control and Nitrogen Laser treated cells, the intensity of the plasmid DNA band have been decreased in Nitrogen dye and Helium-Neo laser treated cells. There was no effect on the toxin production of enterotoxigenic Esch. coli after the exposure of Nitrogen, Nitrogen-dye and Helium-Neon laser. Coagulase production of Staphylococcus aureus was inhibited after 40 minutes exposure of Nitrogen Laser. Manitol fermentation by Staphylococcus aureus became negative after 35 minutes exposure of Nitrogen laser but became positive after 40% 45 minutes of exposure. Since we have to complete the project within one year we could not take up the study of Argon and CO2 laser treated cells.

Immune response specific to outer membrane proteins (OMPs) of Salmonella typhi was studied in vitro in 30 bacteriologically proved cases of typhoid fever, and follow-up after 3 months alongwith 15 age and sex matched normal healthy controls. Cell mediated immunity (CMI) against sy. typhi was studied by leucocyte migration inhibition (LMI), lymphocyte transformation test (LTT), production interleukin-1 (IL-1 alpha) and IL-2, production of leukotriene B4 (LTB4) and LTC4. Enumeration of percent and absolute peripheral lymphocyte sub-populations was performed using flourescein-isothiocyanate (FITC) conjugated mouse monoclonal antibody. Results showed that S.typhi induced specific CMI response in typhoid, both in acute phase and on follow up. Lipopolysaccharide present in OMP prepaations induced only transient cellular response, that too in acute phase of typhoid. Transient non-specific suppression of CMI was observed in acute phase. OMP specific antibodies were detected in all typhoid sera in acute phase as well as on follow up.

The study was carried out for evaluation of various serological procedures for detection of mannan and cytoplasmic antigens as also antibodies for diagnosis of invasive candidiasis. Mannan and cytoplasmic antigens were prepared from the blastosporesand mycelial phases of Candida albicans serotype A strain ATCC-1065 manintained on Sabourauds dextrose agar. Cytoplasmic antigens were purified from mannan fraction by affinity chromatography. Antisera were raised in rabbits against these antigens for antigen detection by latex agglutination test (by coating latex particles with hyperimmune IgG fraction) and countercurrent immunoelectro-phoresis (CIEP). Cytoplasmic antigen fractions were also screened for immunodominant antigens by immunoblot procedure using sera from five patients with histopathologically proven invasive candidiasis. A 47 kDa protein was foundto be the most immunodominant fraction. One hundred patients (confirmed invasive candidiasis-29, suspected invasive candidiasis-30, superficial candidal colonication and without any deep seated candida infection-41) were screened for antigen and antibody. Amongst different candida antibody detection procedures (like immunodiffusion test, CIEP, whole cell agglutination, and immunoblotting assay), immunoblot test for detection of antibody against 47 kDa protein fraction was found to be the most ideal procedure with a sensitivity of 79.3 per cent, specificity of 85.4 per cent, positive predictive value of 79.3 per cent, negative predictive value of 85.4 per cent, and efficacy of 82.9 per cent. This procedure could detect antibody in patients with immunodeficiency. Amongst antigen detection procedures, latex agglutination test for detection of mannan or cytoplasmic antigen was found to be better than CIEP. However, this procedure requires to be evaluated in patients with other fungal infections. Candida specific precipitating antibody detection by gel diffusion or CIEP was of high specificity but lacked sensitivity. Candida whole cell agglutination for detection of antibody was found to be a good sensitive (75.9%)procedure but the specificity was very low (31.7%). There was no significant difference observed in results using either of the phases of Candida - blastospores and mycelial. It may be concluded that detection of specific 47 kDa fraction antigen in serum seems to be a better procedure compared to the antigen detection tests in use currently.

Cervico-veginal flora in pregnancy and its relation with perinatal outcome: The study was carried out to determine the prevalence of Chlamydia trachomatix, Mycoplasma hominis, Ureaplasma urealyticum, Gardnerella vaginalis, Neiseria gonorrhoeae and other aerobic and anerobic organisms in pregnant women and the effect of these on perinatal outcome and maternal morbidity. An effort was also made to evaluate the effect of treatment on perinatal outcome and maternal morbidity. Initially 200 apparently healthy pregnant women attending the antenatal clinic were screened twice for crevical and vaginal flora, once in the second trimester and once in the late third trimester. These women were followed up intrapartum and postpartum. Women with history of recent antibiotic use, obstetrical complications like APH, toxaemia, gestational diabetes, multiple gestation, etc. and those with severe anaemia, diabetes, chronic hypertension, renal diseases, etc. were excluded. The women(116) who were positive for any pathogenic organism formed the study group while those (84) with no evidence of pathogenic organisms served as controls. In order to evaluate the effect of treatment on perinatal outcome, 59 women of a group of 92 screened in the later part of the study were treated. For candida the treatment comprised cotrimoxazole pessaries for the women and candid onitment for their huubands; erythromycin stearate 500 mg 6 hourly for one week was given to both wife and husband for the other pathogens. Initially C, trachomatis, M. Hominis, U. urealyticum, and G. vaginalis were detected in 5.0, 8.5, 28.5 and 15.5 percent women respectively. The prevalence of Candida sp and Gruop B streptococci was found to be 18.0 and 0.5 percent. Lactobacilli alone were present in 42.0 percent, streptofaecal is in 1.5 mobiluncus in 0.5 and Staphylococcus epidermidis in 9 percent of women. Anaerobic organisms were detected in 4.5 percent, staphylococcus diphtheroids and acinetobacter in 1.0 percent each and Escherichia coli in 0.5 percent of women. Sixty eight (58.62%) women had single pathogen while 48 (41.38%) had multiple organisms in various combinations. The incidence of premature rupture of membrane (PROM) was slightly higher in the study group (10 out of 116 : 8.62%) as compared to control (5 out of 84; 5.24%). The incidence of preterm labour was significantaly more in study group compared to control. There was no significant difference in the mean birhtweight of neonates in patients compared to control. Evidence of vertical transmission of infection ie neonates being colonized wht the same organism as the mother, was seen in 18.96 percent of the cases (22 of the 116). Clinical manifestations of neonatal infection in the form of oral thrush was significant difference was seen with conjunctivitis, pneumonia and septicaemia. Post partum fever occurred in 7.5 percent (9 out of 116) women in the study group and 5.95 percent in the control group, but this difference was not significant. Treatment resulted in the lowering of incidence of PROM, preterm labour, incidence of low birth weight, oral thrush in neonates and postpartum fever in mothers.

Characterization of the antigens of Helicobacter pylori and the specific serological response in asymptomatic colonization and infective upper gastrointestinal diseases. The study carried out on endoscopic gastric biopsies of patients with various gastrointestinal disorders to isolate, characterze immunologically and stdy the strain variation in antigenic structure of Helicobacter pylori. A total of 555 endoscopic gastric antral biospy samples from 504 consecutive patients with dyspeptic symptoms were studied for their H. pylori status. Smear microscopy, rapid urease test and culture isolation revealed 46,43 and 23 percent positivity respectively for H. pylori. SDS-PAGE protein analysis of 20 H. pylori clinical isolates consistently having nine major protein bands (molecular weights 90, 65, 60, 48, 35, 31, 28 and 21 kDa). There were detectable strain variations limited to the less prominant bands at 48-68 (1 band), 35-42 (3-4 bands) and 21-28 (3 bands) kDa regions. Western blot enzyme immunoasay (WB-EIA) of electrophoretically separated proteins of clinical isolattes of H. pylori and the reference strain using hyperimmune rabbit serum showed major common immunoreactive components at 21, 28, 42,48, 65, 70 and 116-120 kDa zones. Some minor immunoreactive bands which showed strain variations were limited to 26-32 (1-2 bands), 30-40 (4-5 bands) and > 120 kDa (1-2 bands). WB-EIA with serum samples from patients and controls showed four reactive bands in the molecular weight range of 45-65 kDa in all subjects irrespective of their H. pylori staus or histopathological severity of the lesion. In conrast 4-5 immunoreactive bands between 21 and 45 dDa were found only in patients with positive H. pylori status and evidence of active gastritis. A total of 120 family members of 40 dyspeptic patients (20 eachh age and sex matched H. pylori positive and H. pylori negative) were screened for seropositivity for H. pylori using an ELISA, developed in house and validated. wenty nine of the 69(42%) family members of the positive and 20 of 51(39%) of the negative group were found seropositive for H. pylori antibodies; with no significant difference between two groups. Seropositivity increased with age from 34 percent at < 20 years of age to > 80 percent in the 6th decade. No family clustering could be found on the basis of seropositivity. There was a strong correlation of seropositivity with upper gastrointestinal symptoms in the family members of the H. pylori positive group.

The study was carried out for identification and characterization of puri- fied antigens from hydatid cyst and for identification of immunoreactive hydatidfluid antigenic fractions by Western blotting which could ultimately be used forthe early diagnosis of hydatidosis and to study the kinetics of immune response in experimentally infected animal model. Hydatid cyst fluid collected from hydatid cysts of naturally infected sheepwas processed for the preparation of crude soluble hydatid fluid antigen by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGEanalysis revealed multiple protein bands of molecular weight ranging from 8 to 200 kDa. Of these, most demonstrable protein fractions occurred as a thick band in the range of 66-90 kDa. Western blot analysis of hydatid cyst fluid with serum samples from patientsof hydatidosis, neurocysticercosis, amoebiasis, intestinal helminthic infection and normal healthy individuals indicated the presence of multiple immunoreactiveantigenic components ranging from 8 to 200 kDa. Of these, 8 to 45 kDa antigenicfranctions were found to be specific for the diagnosis of human hydatidosis. Sensitivity and specificity of enzyme linked-immunosorbent assay (ELISA) for thedetection of specific hydatid antibodies in optimally diluted (1:1280) serum samples of patients clinically suspected to have hydatidosis was found to be 81.8 and 100 per cent respectively. Experimental infection could be established in mice by intraperitoneal inoculation of 2000 protoscolices of hydatid cysts obtained from naturally infected sheep. Kinetics of immune response to the crude soluble extract anti- gen revealed positive response by the second month post infection which persis- ted till the sixth month post infection (follow up for six months). Crude soluble extract antigen was found to be more suitable than the purified anti- genic fraction for assessing immune response in early hydatidosis.

Plasmid fingerprinting was applied for tracing the source of nosocomial infection with plasmid containing bacteria. In addition, bacterial chromosomal fingerprinting after restriction endonuclease digestion was done to determine the relationship between isolates of bacteria causing nosocomial infection but not containing plasmids. Forty seven strains of Klebsiella pneumoniae were subjected to klebocin typing by scrape and streak method using 6 indicator strains. All the strains were found to be positive to klebocin thereby offering no discrimination. By klebocin extraction method these 47 strains could be grouped into 11 distinct klebocin types. However, on repeated testing only 35 strains gave similar pattern indicating a reproducibility of 74.5 per cent. Three major categories of klebocin types predominated even in epidemiologically unrelated strains making discrimination difficult. The two outbreaks (August 1994, June 1995) investigated were caused by a klebocin type designated 444, a group indi- cating resistance to klebocin produced by all six producer strains and essentia-lly untypable by klebocin typing. Whole cell protein analysis (WCPA) by SDS-PAGEcould subdivide the 7 strains from the first outbreak into 5 groups while the 3 strains from the second outbreak were found to be homogenous. Ribotyping of thesame isolates indicated 5 groups in the first outbreak and 2 groups in the second outbreak were found to be homogenous. Ribotyping of the same isolates indicated 5 groups in the first outbreak and 2 groups in the second outbreak were found to be homogenous. Ribotyping of the same isolates indicated 5 groups in the first outbreak and 2 groups in the second. Ribotyping had better discrimiantion and indicated that these outbreaks were actually pseudo-outbreaksand not common source outbreaks as was implied by the use of klebocin typing. Forty two strains of Pseudomonas aeruginosa isolated from bronchoalveolar lavage from intubated patients between December 1993 to June 1994 were also ana-lysed. Pyocin typing of these strains by scrape and streak method using 22 indicator strans revealed excellent discrimination in 40 strains. But on repea-ted testing the group designation changed indicating that the system had low reproducibility. SDS-PAGE of whole cell protein profile indicated presence of 45 protein bands of different molecular weights, individual isolates had 37 to 42 prootein bands ranging from 340 to 143 kDa. On the basis of Dice Index ofsimilarity the strains could be grouped into 20 types. Since all strains could be typed the system had adequate typability. Similar results were obtained on repeated testing indicating good reproducibility. Ribotyping of these isolatescould group them into 18 groups; the major group comprised 8 isolates. It is thus clear, that pyocin typing had excellent discrimination but poor to no reproducibility while klebocin typing had adequate reproducibility but notsufficient discimination. Therefore, these methods cannot be of use for reliable fingerprinting of isolates leading to tracing of the source of infection in an outbreak. Whole cell protein analysis by SDS-PAGE offered a relaible, reproducible and fairly discriminating method for identification of clones of bacteria. Plasmid analysis was not found to be a very discriminatory or sensitive indicator for fingerprinting. But ribotyping, using a DIG labelled 18 mer oli- gonucleotide probe complementary to a conserved region of the 16 S rRNA gene showed good results. The probe was stable and hybridized with DNA extracted from a wide variety of bacteria. Since the rrn operon is present in multiple copies in the DNA of each bacteria, a number of bands would light up making determination of the clonal nature of the isolates easy. Discrimination could be improved by selection of more than one restriction endonuclease for cutting the bacterial DNA.

Prevalence of Mycoplasma pneumoniae in acute lower respiratory tract infections. The study was carried out in 249 patients (192 children <10 yr. of age, 43 adults between 18-30 Yr, and 14 elderly >60 Yr) with acute lower respiratory tract infections and 55 normal healthy individuals (matched for age) to find out the age related prevalence of Mycoplasma pneumoniae and correlate the clinical findings with M.pneumoniae to identify the risk factors. M.pneumoniae was found to be an important respiratory pathogen in patients of all ages; the prevalence being 34.89%(67/192) in children,46.5%(20/43) in adults and 14.28%(2/14) in the elderly. Coexistence of M.pneumoniae with other bacterial pathogens (both Gram positive and negative) was frequently seen in all age groups; 17.07,48.9 and 14.28% in children, adults and the elderly, respectively. A past history of parents having had upper and lower respiratory tract infections was identified as a risk factor in the children whose parents had at least 1-3 episodes of upper and lower respiratory tract infections per year. The high incidence of M.pneumoniae in patients of all age groups thus emphasizes the need for examination of specimens of this pathogen in all patients with acute lower respiratory tract infections. For the complete diagnosis of M.pneumoniae infection culture/antigen detection must be supplemented with serological examination so as to plan appropriate treatment strategies for the management of acute lower respiratory tract infections in these patients. Since Mycoplasma infection was seen along with other bacterial p pathogens, it is suggested that the therapy should be directed towards both pyogenic and a typical pathogens for a good clinical and therapeutic response.

The study was carried out to identify and purify immunodominant antigens from circulating antigens of Aspergillus fumigatus present in invasive aspergilliosis as also to evaluate the purified immunodominant antigens in antibody and antigen detection tests for the diagnosis of invasive aspergiolosis in experimentally infected mice and in patients suffering from invasive aspergillosis. Crude antigens of the two stains used in the study ( standard strain Aspergillus fumigatus ATCC 13077 and clinical isolate A.fumigatus MCC LS 77,0010) were prepared and rabbit hyperimmune sera raised against the respective crude antigens (CF-73, MYC-73 and C-10) were used for detection of circulating antigens. Acute invasive aspergillosis was established in cyclophosphamide treated adult Swiss albino mice After infection 100% mortality was observed in mice within four days of infection. After the establishment of experimental animal model of acute invasive aspergillosis circulating anitgens were detected by Western blotting. Prominant protein bands were found at 18 kDa positioin using anti CF-73 antibody.

Enteric bacteria are known to trigger reactive arthritis in HLA B 27 positive individual juvenile idiopathic arthritis (JIA) an immuno inflammatory arthritis of childhood has an association with HLA B 27. Thus we looked for role of enteric bacteria in patients with JIA. Synovial fluid was obtained from 28 patients with JIA and 10 patients with proliferation assay was done using crude lyaste of enteric bacteria Salmonella typhimurium, Yersinia enterocolitica, Shigella flexneri and Camphylabacter jejuni as antigen. Escherichia coli crude lysate was used as a control antigen. HLA B 27 typing was done by PCR using sequence speicific primers.homing of gut educated T cells by tricolor flow cytometry. To see the presence of bacterial DNA16 Sribosomal gene PCR and bacterial species speicific PCR and bacterial species specific PCR was done. Twenty three out of 28 patients were HLA B 27 positive. Synovial fluid cells from 14 of the 28 patients showed antigen specific response, 6 to Salmonella four to Campylobacter two each to Shigella, Yersinia. One of the 10 SF from patients with RA showed response against salmonella antigen. Expression of CD 103 was found to be higher on synovial fluid lymphocyte as compared to peripheral blood lymphocytes in paitents with JIA. Expression of CD 103 was higher on CD8+ T cells as compared to CD4+ T cells. Bacterial DNA was present in majority of samples in the synovial compartment as well as in peripheral blood. Bacterial species specific PCR was begative for all samples except one sample. In one patient with JIA sample were positive for Salmonella typhimurium DNA. To complete, the data indicate that enteric bacteria play a role in disease exacerbation in patients with JIA. These patients show evidence of selective homing of gut-primed cells in the synovial compartment. Our Our findings of specific cellular immune responses against enteric bacteria in a proportiion of patients with JIA suggests that despite absence of history of a symptomatic enteric infection , these patients have a disease resembling chronic ReA. Thus a proportion of patients with JIA may have a forme fruste of chronicReA.

Accurate identification of Genus/species of the fungi in the tissue section subjected to histopathological examination is important for the institution of speicfic therapy. Tissue forms of several fungi have similar appearance. Sometimes it is differentiate the fungi accurately in the tissue sections based on the morphological forms. So the present study was undertaken with the objective to develop fluorescent antibody reagents to differentiate some morphologically closely resembling fungi in the tissue sections such as Fusarium, Aspergillus and Pseudallescheria boydii and similarly betewwn yeasts like Candida and Cryptococcus. Antiserum was successfully raised against Aspergillus funigatus, Aspergillus niger, Aspergillus, Fusarium oxysporum,C. albicans, C. krusei, C.tropicalis, Cryptococcus neoformans and Pseudallescheria boydii. The cross reacting antibodies present in the antiserum was removed by adsorption with the cross reacting antigen/ antigens. However it was not possible to remove cross reaction between species under the Candida or Aspergillus. Initally all antiserum were evaluated by the indirect Immunofluorescence (IIF) test. Adsorbed pooled Aspergillus antiserum and Fusarium antiserum specifically identified the respective fungi in the mice tissue sections by IIF test. Both Cryptococcus neoformans and candida antiserum sepcifically recognized the respective yeasts in the mice tissue sections by IIF test. Direct immunofluorescent (DIF) reagents were prepared by purification and labeling the specific antiserum with the fluorescent isothiocyanate (FITC). Labeling was unsucessful of P. boydii antigen. The specific reagents were tested with the mice tissue sections containing specific fungi. The reagents prepared against C. albicans, Aspergillus, Cryptococcus neoformans and Fusarium oxysporum specirfically identified the respective fungi from all the mice tissue sections tested indicating that these reagents may be valuable in differentiating the fungi in the tissue sections of the patients. C. albicans, Aspergillus and C. neoformans reagents were also tested with the culture and/or smear proven tissue sections from patients sample contining the specific fungi. All these reagents gave the fluorescence ranging from 3+ to 4+ with the tissue sections of patients tested indicating that these reagents can specifically identify the fungi in the tissue sections processed for the histopathological examination. Though Fusarium reagents gave positive results with the tissue sections of mice origin, we could not test it on human tissue section due to non availability of such confirmed case in histopathology department. We now propose to evaluate these reagents in other centers. At the time of institution of this project, it was decided that Prof. S. Dutta Gupta of Histopathology Department, All india Institute of Medical Sceinces would evaluate the reagents in his pool of specimens.

The aim of the study was to determine the microbiological profile and antibiotic susceptibility patterns of organisms isolated from diabetic foot ulcers of different Wagners grade. Potential risk factors for infected ulcers with methicillin resistant Staphylococcus aureus (MRSA) and outcome infections were also studied. Pus sample for bacterial cultures were collected from 100 patients admitted with diabetic foot infections over a period 3 years. Sixty-two had co-existing (62%) osteomyelits. Staphylococcal isolates were tested for susceptibility to oxacillin by screen angar method, disk diffusion and mec A based PCR and susceptibility to vancomycin by screen agar. Gram negative bacilli were testedc for extended spectrum beta lactamse (ESBL) by double disk diffusion. Potential risk factors for MRSA- positive samples were explored. A total of 224 pathogens were isolated ; 187(89.5%) aerobes and anaerobes 37(16.5%). Gram-negative aerobes were most frequently isolated (48%) followed by gram-positive aerobes and anaerobes (32.2% and 10.2%, respectively). Methicillin- resistance and ESBL production was noted in 60.5% and 41.6% of bacterial isolates. MRSA status is not associated with patient demographic characteristics, ulcer type, ulcer duration, ulcer size or duration of hospital stay. The factors significantly associated with MRSA positive status were presence of hypertension (p= 0.001), neuropathy (p=0.003) and esteomyelitis (p=0.02). Significantly more number of patients with MRSA infections required amputation (p=0.002). It is concluded that infection with MRSA is common in diabetic foot ulcers and is associated with increased requirment for amputations. These results can guide appropriate empirical antibiotic therapy in these patients and reduce the risk of complications

Eighty percent of the cases of acute exacerbation of chronic obstructive pulmanary disease (AECOPD) have an effective etiology; atypical bacteria including Mycoplasma pneumoniae account for 5 - 10% of these. However, some studies could not find evidence of association of M. pneumoniae with episodes of AECOPD at a referral hospital in Delhi. Sputum samples and throat swabs were collected from a total of hundred patients of AECOPd attending Vallabhbhai Patel Chest Institute and thirty five healthy control subjects. The samples were investigated for the presence of aerobic bacterial pathogens and Mycoplasma pneumoniae. Diagnosis of infection with Mycoplasma pneumoniae was based on culture serology, PI antigen detection and PCR for the PI antigen detection and PCR for the PI gene. Bacterial etiology could be established in 16 patients. The organism isolated most frequently was Pseudomonas from 4, Klebsiella sp. from 2; and Acinetobacter sp. and Moraxella catarrhalis from one case each. In three of these cases, the M. pneumoniae infection. However, one of the cases which was cultrue positive for Pseudomonas sp, also had m. pneumoniae specific 1gG titer>= 100 U. This patient, thus probably and a mixed infection of Pseudomonas sp and M. pneumoniae. Of the100 patients studied,39 were IgG positive. Ten of these showed boderline positivity.Of the thirty patients followed up, 5 of showed a four fold rise of antibiodies in 4 weeks. Amongst the 18 samples positive with GPA, three samples had a titers>= 1;320. The IgM ELISA assay in our study was positive in 4 serum samples. IgA antibodies were seen in 10 patients. None of the 8 borderline positive patients which followed up showed a significant change in IgA titer. Four patients had both positive IgG and positive Iga titers, indicative of a primary . pneumoniae infection. P1 antigen was deteced in 3 samples. Two of these patients had positive IgG and IgA responses, but no IgM responses. PCR for the P1 adhesin gene was negative in all the samples studied. Teh prevalence of M. pneumoniae infection in our study population was significantly higher than that in the control group. To conclude, on the basis of derology and antigen assay, at least 17% of the 100 patients of the exacerbation of COPD showed evidence of M. pneumoniae infection. Mycoplasma pneumoniae, therefore, emerged as an important etiological agent of AECOPD in our study population.

The study was carried out to understand the biochemical mechanisms which regulate the cyclic nucleotide metabolism in rat brain and physio - logical importance of this regulation. The effects of factors modulating adenylate cyclase which are not related to neurotransmitters, were studied. The possible influence of biogenic amines like dopamine (DA), serotonin (5-HT) norepinephrine(NE) and also acetylcholine (ACh) and other agents on adenylate cyclase (AC) and phosphodiesterase (PDE) in the rat brain was also evaluated. Dopamine NE and 5-HT stimulated rat brain AC activity A dose dependent stimulation of AC by DA (with 1-1000 muM) and NE (with 25-75muM) was observed with DA being more potent. Serotonin stimulated rat brain AC activity by 60 per cent at 10 muM, which increased to 100-160 per cent in presence of NaCl. But the dose dependent stimulation of AC by 5-HT was not as sensitive as by DA and at higher concentrations(more than 10 muM) inhibited AC. The presence of acetylcholine inhibited AC activity. Higher concentra- tion of ACh inhibited the DA sensitive AC and this inhibition reversed by increasing DA, thereby showing a competitive inhibtion of AC by ACh. It is thus probable that the action of biogenic amines in central nervous system (CNS) may be mediated (atleast in part), by cAMP formed in the post synapt receptor sites. Atropine and scopolamine both muscarinic receptor antagonists prevented the inhibition of AC by ACh without affecting basal activity. The action of atropine on DA sensitive Ac could be reversed by increasing the concentra- tion of DA. The stimulation of AC was effectively blocked by antipsychotic drugs known to block DA receptors. Phosphodiesterase (PDE) showed two Km(Kinetic constant) values; a high Km 0f 0.5x 10^-3 M and a low Km of 3.8x10-6M. Theophylline was found to be a most potent inhibitor of PDE with a inhibitor constant (Ki) of 1x10-4M. Both dibutyryl cAMP and isobutyl methyl xanthine (IBMX) also inhibited PDE with Ki of 1.2x10-6M and 2x10-4M respectively. Copper and zinc inhibited PDE whereas in absence of Mg both Ca and Li could activate PDE. Antipsy- chotic drugs like fluphenazine, mepazine being more potent. The relative potency (in decreasing order) being mepazine>trifluoperazine>phenothiazine> fluphenazine>haloperidol. It was observed that both calmodulin and calcium were required for the activation of PDE. Both Ca and calmodulin were found to stimulate Ca sensitive ATPase in synaptosomal membranes. The activation by calmodulin was dose dependent and 5 muM CaCl2 was optimal concentration for maximal activity of Ca ATPase in-synaptosomal membranes. It was concluded that a neurotransmitter sensitive AC is present in rat brain ; DA sensitive being more potent than NE and 5-HT sensitive AC. DA receptor appeared to be associated with AC in rat brain. It was established that ACh was a most potent competitive inhibitor of DA sensitive AC. DA and ACh act synergistically on AC. It appeared from the study that AC system may be a useful in vitro model for study of the functional role of DA and ACh receptors and the pharmacology of certain drugs.

Certain basic immune functins in intracranial tumour patients. In order to elucidate the role of neuralmodulation of immune response in patients of intracranial tumours, immune parameters were studied before and 30 days after treatment in these patients. Normal individuals matched for age, sex and socio-ecomomic status served as controls. Patients with benign tumours were treated by surgery whereas those with malignant tumours were treated by surgery whereas those with malignant tumouurs were treated by surgery, radiotherapy and chemotherapy. It was found that patients with benign and malignant tumours had higher total leukocyte count before treatment which decreased significantly after treatment. Lymphocyte percentage showed a reduction but absolute monocyte count and monocyte percentage showed an increase in both benign and malignant tumour patients. There was an elevation in neutrophil functions as evidenced by increased phagocytic index, avidity index, NBT (Nitroblue tetrazolium) reduction and SIC (serum soluble immune complex) index in brain tumour patients. As compared to controls total T cell and T4 counts were reduced in both benign and malignant tumour patients before and after treatment whereas there was an increase in T8 counts. Serum IgA and IgG levels showed a decrease and IgM level an increase. Preoperatively altered immune parameters like lymphocyte percentage, monocyte percentage, absolute T lymphocyte count, T8 count and IgA levels returned to base- line levels after treatment in benign tumour patients. The study confirmed the altered innate, cell mediated and humoral immunity in brain tumour patients before and after treatment. Most of the immune changes observed were related to cell mediated immunity ad were immunosupperssive. Though the pitutary region tumours are benign, patients with these tumours showed either marked increase or decrease in the levels of immune parameters studied than observed in those with tumours in other areas. The immunomodulatory effects of malignant tumours were more marked than those of benign tumours.

Study of the natural history and serizure outcome in patients with solitary cyticercus granuloma and seizure. Aprosepective study was carried out in 183 patients with seizures to determine the natural history of solitary cerebal cyticercus granuloma (SCG) and seizure outcome after the resolution of the granuloma and simultaneous or early withdrawal (2 to 3 months after resolution) of antiepileptic drags. Only those patients who fulfilled all the diagnostic criteria (clinical and computerised tomographic for SCG) were included in the study. One hundred and twenty nine (70.5%) pateints presented with patial seizures with or without secondary generalization, 53 (29%) with generalized seizure and 1(0.5%) with partial complex seizure. Of these 78.1% patients had 5 or less seizures, 16.9% had 6-10 and 4.9%>10 seizures at initial presentation. Most of the patients had their initial CT scan within one month of their first ictus. Majority (93%) of patients could be managed with a single antiepileptic drug (monotherapy). Breakthrough seizures occurred in 30(16.4%) patients who needed increased dose of the drug or an additional drug. ELISA for cysticercus antibodies in serum was of limited value as it was found to eb negative in a majority of patients. The CT lesions were typically between 5-15 mm in maximal dimension and were usually associated with mild to moderate oedema. The parietal love was the most favoured site of lodgement of the granuloma. SCG was found to have a varied natural history and the granulomas resolved spontaneously though at different rate in individual patients. Kaplan meier analysis of lesion resolution in the present study showed that at 3 months after first CT scna, 96.7% patients had persistent granulomas (though smaller in size in nearly 40% patients ), at 6 moths 75.5% patients and even after 12 months 46.5% patients had persistent granulomas. The shortest period recorded for the resolution of lesion was 64 days after the initial CT scan whereas the longenst duration of persistent lesion seen in the CT scan was 429 days. Antiepileptic drugs were withdrawn within 2-3 months, after resolution of granuloma was seen on the CT scan. O the 60 patients who could be followed after withdrawal of the drugs ( median follow up period 10 months), 57 were symptom free, and recurrence of seizures was noted in 3 patients at 3 to 10 months after withdrawal of drugs. It can be concluded thath SCG, a common cause of seizures in Indian patients, is a spontaneously resolving lesion with a variable rate of involution ranging from weeks to a years or more. The first follow up CT scan can be delayed for up to 6 months decrease the number of scan examinations. Patients with SCG generally have a bengin seizure prognosis aqnd the antiepileptic drugs can be withdrawn after resolution of granuloma. However, a longer follow up is necessary to arrive at a definite conclusion regarding the longterm seizure outcome in these patients.

Effect of low level prenatal and neonatal irradiation on the adult brain development and reproductive function is mouse. The study was carried out on inbred Swiss albino mice to evaluate the effect of gamma irradiation at selected critical developmental stages during gestation on the adult neurophsiology and to see, if the neurophysiological effects of prenatal irradiation persist to later part of life. Efforts were also made to find out if prenatal irradiation would impair the reproductive performance of mouse. Pregnant mice were whole body irradiated with 0.25,0.35 or 0.5 Gy of gamma radiation on days 11.5, 12.5 (late organogenesis days), 14.5 or 17.5 (foetal days) post coitus (pc) and allowed to complete gestation and parturate. Behavioural and haematological changes were studied in the adults at 6, 12 and 18 months of age. Reproductive performance was tested at 6-8 months of age after mating the irradiated F1 males with normal females of the same age. For neonatal exposure, pups were whole body exposed to 0.5 Gy of gamma radiation on days 4, 5 or 6 post-partum. Adult behaviour was not affected by prenatal irradiation with 0.25 Gy. Exposure to 0.5 Gy on days 11.5, 12.5 or 14.5 pc produced significant changes in the exploratory, locomotor and learning and memory performances at 6 months of age. The changes were more pronounced after irradiation on the late organogenesis days (11.5 and 12.5 pc) than on the foetal day (14.5 pc). Normal brain function was restored by 12 months in all animals, except those exposed on 11.5 day pc which the impaired locomotor activity and learning performance of the young adult mice. Animals born to mice irradiated on day 17.5 pc werre more resistant to these effects. The peripheral RBC counts and haemoglobin showed significant decrease at 6 months of age when the pregnant mouse was exposed to 0.5 Gy on the foetal gestation 914.5 or 17.5 pc) days, but not during the organogenesis period. Post-partum exposure did not have any significant effect on the adult behaviour or haematological parameters. None of the hippocampal biogenic amines (noradrenaline, dpamine, 5-hydroxytryptomine and its metabolite -5-hydrooxyindolacetic acid) studied showed any significant change after any level of exposure on any of the gestation days. The reproductive performance of the F1 males exposed to different doses of irradiation did not show any significant change from normal. It is clear from the study that the late organogenesis period, especially day 11.5 pc is a particularly sensitive phase in braain development and long lasting behavioural changes can be produced in the adult by exposure at this stage to doses below 1 Gy.

This study is about dairy products such as milk and ghee contain saturated lipids and cholesterol which are considered to be atherogenic in nature. However an earlier studies indicated that feeding rats a diet containing dairy lipids exhibited hypocholesterolemic effect (1). Studies conducted under similar conditions with diets containing egg yolk lipids did not show any benefical effects in modulating serum lipid profile in rats. These studies were conducted in normal diet. However cholesterol enriched diet are known to elevate serum cholesterol levels and the influence of diary lipids under hypercholesterolemic conditions is known. To evaluate this, the present investigation was undertaken to study the influence of diary lipids and egg yolk lipids in rats rendered hypercholesterolemic by feeding a diet enriched in cholesterol. The hypocholesterolemic properties of dairy fat cross examined when rats were rendered hypercholesterolemic by feeding a diet containing. Supplementing the diet with cholesterol at 0.5% level incrased serum cholesterol level cholesterol level by 40% compared to rats fed diet without cholesterol. Rats fed diet containing milk or ghee lipids had 9-11% lower levels of serum total cholesterol compared to control rats fed with cholesterol enriched diet but devoid of milk lipids. The present experimental observation indicates that milk lipids, either in normal cholesterolemic cnditions or in hypercholesterolemic condition will lower serum cholesterol concentration in rats. It is tharefore concluded that diary lipids can beneficailly modulate serum and tissue lipids and hence it is not a cause for concern in terms of increasing risk factors for cordiovascular diseases.

Combined Antidiabetic effect of Sodium orthovanadate and other antidiabetic compounds such as Trigonella seed powder and Momordica fruit extract a new propective therapy: Hyperglycemia druing diabetes results in majority of the metabolic derangements and clinical complications that contributes towards the morbidity and mortality of diabetic patients, the latter may be prevented or reversed with an effective control of increased blood glucose. Repeated insulin administration controls the short term diabetic symptoms, but it fails to prevent the serious vascular and other complications of diabetes. Moreover, exogenous insulin treatment fails to provide a regulated glycemic conditions in association with variable dietary intake and variable physical activity in Type 1 diabetes. Episodes of severe hypoglycemia leading to a deleterious cerebral impact are common during tinsulin administratioin (30). Other drugs available also have similar problems in addition to their toxic side effects. Therefore, appropriate antidiabetic compounds without or less toxic effects are required to primarily control the blood glucose, which can subsequently, effectively reverse the altered biochemical and physiological changes during diabetes. The insulin mimetic action of vanadate has been explored since the last decade in various diabetic model systems including IDDM and NIDDM juman subjects (31). Trigonella seed powder and Momordica charantia fruits are well known antidiabetic agents and have been shown to rejuvenate and Beta cells in the islets of Langerhans, thus increasing the capacity of insulin secretin in type 1 diabetes (32). The insulinotropic property of 4 hydroxyisoleucine, and amino acid extracted from trigonella is also suggestive of insulin secretion modulation in its therapeutic action (33). Similarly insulin like peptide (polypeptide-P) has been isolated from Momordica fruits and has been shown to act like insulin (34). However, no detailed study is available to establish whether the plant extracts follow similar signaling biochemical routes such as as those taken by insulin and vanadate. Being a natural product and a part of regular diet with multitude of antidiabetic effects, Trigonella seeds and Momordica fruits can possibly be used as insulin replacement or adjuvant in the management of diabetes. Vanadium has a poor therapeutic index inspite of its significant biological potential and well known insulin mimetic mechanism (13). However ligands, when complexed with vanadium, potentiate its insulinomimetic acitivity, both in-vivo and in-vitro. Many perspective involving biochemistry and bioinorganic chemistry of vanadium and its complexed with several types of ligands have been proposed as useful for treating diabetes mellitus in experimental diabetic animals. It has been shown by Shinde et al, 2001(35) that chronic treatment with an organic complex of vanadium such as bis(maltolato) oxovanadium (IV) (BMOV) was effective in improving glucose and lipid homeostasis. Hence, the attempt and emphasis on the use of vanadium complexes in treatment of diabetes melliuts is emerging as a new concept. Earlier works from our laboratory and in the present work an attempt has been made to prevent the toxic effects of vanadium by reducing the dose of vanadium administered (0.2 mg/ml) and combining its administration with that of plant products for the treatment of the diabetic animals. This combined treatment of vanadium with plant products was observed to be more beneficial with respect to biochemical stablization as well as improvement in the physiological parameters of experimental diabetic animals, besides establishing normal glucose and lipid homeostasis. It can be suggested that there may be some in-vivo formation of organometallic compounds by sodium orthovanadate with organic compounds, may be active constituents, made available by plant extracts that are responsible for bringing the improved management of blood glucose level and at the same time also reducing the toxicity of sodium orthovanadate. The findings of the present work suggest the effectiveness of the combined therapy of vanadate and plant products on the control of glucose homeostasis and lipid metabolism druing experimental diabetes and can be sonsidered as a better alternative for the amelioration of diabetes. In the present study diabetes was produced by ginving alloxan injection to the rats, all diabetic rats showed severe hyperglycemia, hyperphagia, polydipsia, glycosuria and severe body weight lose. After giving treatment with insulin, trigonella, Momordica, higher dose of vanadate alone and lower dose of vanadate with trigonella and momordica restored high blood glucose level to the normal values. Combined treatments were found to be much more effective in normalizing the diabetic complications in short and long term diabetes. However, further investigation compounds formed in-vivo, to find the active constituents of these plant products and to work out their exact molecular mechanism in combination with sodium orthovanadate.

Several chemical compounds like captopril, Analapril etc are employed as antihypertensive drugs, which are known ACE inhibitors. The most common adverse effect associated with ACE inhibitor is a presistent non productive cough. Hence there have been attempts to develop ACE inhibitors sans the side effects. Recently several ACE inhibitory peptides have been isolated from plant foods. Enzymatic digensts of various food materials, including casein, zein, gelatin, sake, soyabean fermented milk, sardine muscle, tuna muscle, dried salted fish, fish sauce and onion. These peptides are reported to inhibit the ACE both in vitro and in vivo studies. However limited reports exist regarding isolation and employment of ACE inhibitory peptides from cereals and pulses. Hence this project mainly aims at Our earlier studies have also shown that chymotryptic digested fractions of Sorghum storage protein, alpha-kafirin, possess strong ACE inhibitory activity. We characterized the inhibition from a kinetic point of view. We determined the IC50 of the potent fractions were found to be 24.3 muegram/ml to 1.3 muegram/ml while the IC50 of Captopril was 0.0067 mueM. The inhibition by teh fractons was found to be competitive and uncompetitive with the substrate. Our results offer a biochemical mode of inhibitino by the hydrolytic fractons were aslo found to inhibit rat tissue ACE (in vitro)to a significant extent. The results also suggest the potential mechanism that might occur in vivo, to act as a potential antihypertensive agent. Studies were conducted to measure the basal angiotensin converting enzyme (ACE) activity in rat tissues. Comparative evaluation of the enzyme activities was carried out in different tissues of rat of different age groups of rats. Heighest ACE activity was seen in lung of adult rats, the major site of its, synthesis. However circulating ACE activity, in serum, was minimal in rats of all the age groups. Cadmium chloride (CdCl2), a well established inducer of hypertension in animal models was found to alter ACE activity in different rat tissues to varying extent. CdCl2 induced a significant increase in ACE acitvity in the rat lung, while it decreased the enzyme activity in serum. Pre treatment with Cap;topril, a known ACE inhibitor was found to reserve the Cd induced alterations in tissue ACE activity. Further in vitro studies reveled tha serum, small intestine and brain ACE were more susceptible to inhibition by captopril. The above studies have clearly yielded steps and procedures for studies employing ACE for its deploymen in evaluation of ACE inhibitory compounds. isolation and further studies on ACE inhibitory activities of peptides from cereals and pulses common to Indian diet in perticular such sorghum, ragi and black gram dal. In our preliminary studies were prepared and evaluated protein hydrolysates from sorghum and rgai flour. Both total proteins as well as storage proteins ( prolamins- the alcohol soluble proteins) were subject to proteolysis with selected emzymes viz. trypsin, chymotrypsin, pronase, papain etc. The digests obtained were analyzed by SDS-PAGE and those showing maximum digestions were subjected to further studies. Trypsin and chymotrypsin digests of sorghum total prolamins and pronaase digest of ragi tatal prolamins were found to yield peptide fractions possessing signicant ACE inhibitory activities (50% and 62% respectively)

Immunocytochemical localization and biochemical estimation of peetide hormone binding capacity of breast tissue. The study was carried out to examine immunohistochemically detectable prolctin (PRL), follicular stimulating hormone (FSH) and leutinising hormone (LH) binding sites in various breast lesions using a highly sensitive modified dinitrophenyl hapten sandwich staining procedure and biochemically estimating PRL binding in particulate membrane preparations of breast tissue. One hundred and seven formalin fixed paraffin embeded breast tissue sections were immunostained for localisation of prolactin binding sites in benign and malignant breast lesions. Of these, 35 and 28 sections were also stained for FSH and LH binding sites respectively. Eighty tow percent of breast carcinoma gave an intracellular positive reaction in tissue sections for prolactin binding sites but the positivity was 72 percent in fine needle aspiration smears. In an attempt to biochemically quantitate prolactin receptors, both purified and crude plasma membranes of tissues from benign and malignant lesions, were prepared. Breast carcinoma showed more consistent staining as compared to benign breast lesions. Also, prolactin receptors could be located biochemically in malignant breast tissue membrane preparations. Prolactin binding sites were present in greater proportion in peri or post menopausal breast cancer patients. It is thus concluded that the more consistent immunostaining in breast carcinoma as compared to benign breast tissues as well as presence of PRL receptor in crude membrane of brest carcinoma tissue suggest the role of PRL in mammary tumourigenesis. The results of immunocytochemistry on FNAC being ecoomical, quick and nonoperative procedure, couls be more easily used for study of hormone binding sites in breast lesions.

The study was carried out to isolate estrogen receptor (ER) protein from ER human breast cancer cell lines (like MCF-7) and also from ER human breast tumours and generate monoclonal antibodies to the purified ER protein. Immunohistochemical studies were carried out on human breast cancer tissues and fine needle aspiration biopsy (FNAB) specimens using monoclonal anti- bodies specific to ER. Estrogen receptor protein was isolated from MCF-7 cell lines and breast tumour tissues by pulverization of the material snap frozen in liquid nitorgen, polytron homogenization, high speed centrifuga- tion. The ER was characterized by Scatchard analysis and sucrose density gradient centrifugation. SDS PAGE identified the ER as a single discrete protein with a molecular weight of approximately 66-67 k Da. Three monoclonal antibodies (MAbs) were generated by hybridization of immune spleen cells of BALB/c mice immunized with ER, with Sp2/O myeloma cells using 50 per cent PEG as fusing agent. The positive clones were screened by ELISA and selected by PAP technique using ER positive frozen breast tumour tissue sections. These MAbs, designated as ER E5, ERE10 and ERD7 were found to recognize a cytosolic ER. These MAbs were purified by ammonium sulphate precipitation and protein A sepharose chromatography Specificity of these MABs to ER was ascertained by dot blot analysis, western immunoblot, immunoprecipitation, radioimmuno uptake studies and immunofluorescence study. Analysis of large number of normal breast tissues, benign and malignant breast lesions of different histological types, breast lesions of different histological types, breast FNAB specimens and other ER tissues of carcinomas of the ovaries and endometrium by ABC and PAP immunocyto- chemistry revealed that these ER MAbs exhibited specific staining of cytosolic ER. Most of the tumour cells were strongly positive in the case of infiltrating ductal carcinomas and mucoid adenocarcinomas. The ER values of 50 malignant breast tumour cytosols determined by DCC assay and by enzyme immunoassay (EIA) using MAb ER E5 were found to correlate. For 15 typical breast tumours with ER values in the range of 3.5-270fmol/mg protein, the staining paterns were proportionate to ER values. It was concluded that MAbs ER E5, ERE10 and ERD7 might serve as a valuable diagno- stic tools. Besides, these may be highly useful in understanding the molecular mechanism by which estrogens stimulate cell proliferation.

The study was carried out for investigating the blood flow patterns and rates in facial artery in patients with cancer cheek and age and sex matched patients with leukoplakia cheek, individuals addicted to tobacco chewing and smoking separately and normal healthy subjects to correlate the pattern of blood flow to (i) histopathological grading of cancer cheek, (ii) responsee to chemotherapy,(iii) occurance of cervical lymph node metastasis and (iv) survival. There was abnormality in blood flow in the facial artery in chronic tobacco chewers in the form of partial obstruction. In cancer cheek also similar findings were noted. In addition the doppler study revealed formation of collatrals in cancer cheek. Khaini was found to be the most common form of addition in 71.42%. The risk ratio (Odd's ratio) due to Khaini chewing was 1.036, to smomking 1.088, and addition of alcohol increased the risk to 1.097. Night quid keeping habit had odd's ratio of 3.73 (P<0.001). Buccal vestibular mucosa was found to be the commonest site of cancer (54.46%). Ulceration and pain were prominent clinical findings in cancer cases. There was severe nutritional derangement in cancer patients (P<0.001). DNA content was significantly raised in cancer patients in comparison to tobacco addict and normal controls.

The study was carried out to determine the frequency of hepatocytic of preneoplastic nature in livers of population groups with different incidences of hepatocellular carcinoma and Wister strain albino rats rhesus monkeys exposed to different hepatocarcinogenic agents. Morphological and cytochemical profiles of preneoplastic cells were also studied. Rats were treated with dimethylnitrosamine(single dose of .4 mg/100 g bodywt. intraperitoneally 24 h after partial hepatectomyl) or acetylaminofluorene (3 doses of 3 mg/100 g body wt. i.p. at 24 h intervals followed by partial hepatectomy 24 h after the last dose). A total of 5 rats and 4 monkeys were treated with aflatoxin b1. Qualitatively and quantitatively the sequence of preneoplastic and neoplastic events were similar in rats irrespective of the carcinogen used. Structurally altered hepatocytes occured usually as small foci which later evolved into larger areas as nodules of altered liver cells. In monkeys treated with aflatoxin B1 changes were mild and the lesions few. Histochemical localization of enzymes of carbohydrate metabolism viz glycogen synthetase glycogen phosphorylase or glucose-6 phsphate dehydrogenase revealed no significant change in these enzymes. No significant changes were also noticed with the other enzymes. The acidophilic cells showed a variable pattern of staining for various enzyme It appears that chemical carcinogenesis induces multifocal hepatic glycogenosis which may be related to development of multicentric tumours. It is probable that metabolic disturbances leading to hepatocellular glycogenosis might be related to the neoplastic transformation of the hepatocytes. Liver sections prepared from livers obtained from medico-legal autopsies of 114 cases (55 from Madras and 59 from New Delhi) were studied for the presence of altered cell foci were observed in 19 (34.5%) cases from Madras compared to 14 (23.7%) in NEw Delhi. The commonest cell type was the clear (glycogen rich) cell and some of these foci had acidophil cells admixed. The number of foci with acidophil cells were more in livers obtained from Madras compared to New Delhi.