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Animal Sciences

The Programme for Animal Sciences aims to promote basic research in organismal biology. During the year, individual projects as well as joint collaborative projects from different institutions were funded. To augment scientific manpower and to facilitate value addition to R&D proposals, training programmes, interaction meetings and SERC schools were organized.

Brain-storming sessions were organized on 'ehelminth taxonomy', and 'eornithological sciences' in India. A brain storming session on 'eHelminth taxonomy: revisited'f, 2nd in the series, was organized at Kashmir University, Srinagar during November, 3-5, 2007, to provide a forum for practicing helminthologists to take stock of state of the affairs of the subject, to identify bottlenecks in the progress of the subject, and to find out mechanism to remove them and also to enable incorporation of new tools and techniques in the field, to make it attractive, informative and competitive globally. Another brain storming session on ornithological sciences in India was organized at Gurukul Kangri University, Haridwar to thrash out activities to promote avian biology.

New research projects were sanctioned in the areas such as faunal diversity, parasitology, reproduction, developmental biology, immunology, neurobiology, ecology etc. A total of 35 projects were sanctioned. In addition to the above, a major project on sleep studies for a period of five years is also being implemented.

Faunal diversity: The projects sanctioned under the programme included Molecular assessment of genetic diversity in the only remaining population of Asiatic lions (Panthera leo persica) in India; Genetic characterization of tunas using DNA markers; Diversity of spiders in Nanda Devi Biosphere Reserve; Taxonomy and biology of entamopathogenic Rhabditis (Oscheius) sp. (Rhabditidae: Nematoda) and its bacterial symbionts; Biosystematics, molecular and ecological characterization of indigenous entomopathogenic nematode biodiversity; Biodiversity of subterranean ground waters of India, with special reference to Copepoda and Bathynellacea (Crustacea).

Parasitology: An ecological study of complex life cycle parasites of wild mammalian carnivores; Gastrointestinal parasitic fauna in the ruminants of Jammu & Kashmir state with emphasis on prophylaxis; Taxonomic evaluation of endemic monogenean parasites of freshwater fishes; and a joint collaborative project on the study of host-parasitoid interactions with special reference to Uzi fly (Diptera:Tachinidae) and Antheraea assama (Lepidoptera : Saturniidae) were the projects sanctioned in the area.

Reproduction: The projects sanctioned in the area included, Differential regulation by testosterone and FSH in rat testis; A study of bidirectional communication between Leydig and immune cells in the testis of wall lizard Hemidactylus flaviviridis; and PCB- induced disruption in the expression of selected genes in sertoli cells of adult rats; Molecular mechanism involved in the regulation of perch (Anabas testudineus) oocyte maturation; Studies on influence of stress and stress related peptides on ovarian functions in the fish, Tilapia mossambicus; Processing of photic signal in the pineal organ and ovarian functions in carp Catla catla; Localization of Nitric Oxide Syntheses and role of nitric oxide in ovarian steroidogenesis and maturation in catfish Heteropneustes fossilis (Bloch.); Molecular mechanism underlying nuclear chromatin condensation.

Developmental Biology: A joint collaborative project on molecular mechanism of action of ultraviolet radiation and lithium during regeneration and pattern formation in Hydra; Role of Basic fibroblast growth factor (FGF2) on epimorphic regeneration; Role of lysosomal enzymes, oxidative stress and hoxb8 gene in regulating of regenerating tail tissue of frog larvae; and an investigation on the role of tumor induced expression of hsp70/ hsp70 peptide complexes on activation of macrophage function and regression of tumor were the projects sanctioned.

Immunology: Studies on the Immune Responses of the Striped murrel, Channa striatus bloch; and Molecular characterization and transcriptional regulation of the Vitellogenin receptor from the cotton pest, Spodoptera litura, were sanctioned.

Neurobiology: Implication of Nitric oxide in Hyperammonemic Neurotoxicity; Importance of cartpeptide in learning and memory traces, reward and reinforcement, and hypophysical regulation in rodents; Isolating, characterizing and cloning suppressor mutations of the kinesin-like C. elegans motor UNC-104; An investigation of the role of retinoic acid signaling in development of the hippocampus; and a major project on Centre for Sleep Studies were sanctioned in the area.

Other projects sanctioned in diverse areas such as, ecology, chronobiology, communication etc., include, Ecology and adaptive radiations of some endangered reptiles and amphibians; Behavioral Ecology of the Rock Lizard, Psammophilus dorasalis; Reduviid salivary toxin and its zootoxic effects on polyphagous pests; Sensitivity of the entrainment pathway in a photoperiodic bird; and Acoustic communication in the Indian Chat Cercomela fusca and the pied bush chat Saxicola caprata: characterization and behavioural correlates of songs and calls.

Achievements
Some of the findings of ongoing and completed projects under the programme are as follows:-

Chronobiology
The studies on the role of melatonin in synchronisation of avian circadian rhythms brought out the observations that it was the first study of the kind undertaken on a Palearctic-Indian avian migrant, and on temporal physiology of the spring migration in the Palearctic-Indian avian migratory system anywhere in the world. The photoperiodic clock is sensitive to light wavelength and light intensity. Bird'fs photoperiodic system has the capacity to discriminate between different light wavelengths and light intensities. The duration of food supply and/or the time of day at which food is available affect photoperiodic stimulation of fattening and gain in body mass as well as the growth and development of gonads in the migratory bunting etc.

The study on Circadian patterns of transcriptional clock genes (per, tim and dbt) at wavelengths of light in several mutants of Drosophila reported a conspicuous indication that the WT and mutants differ widely when they were exposed to different regimes of light-dark schedules leading to the conclusion that blue light photoreceptor, cryptochrome is very much essential for the synchronization of the clock. Further, defective organs, such as, greatly reduced wings, as in the case of vg mutants would lead to poorer locomotor activity when compared to wild type. The weaker expression patterns of per and tim I Cryb flies suggested the significant role of blue light photoreceptor, cryptochrome for a stronger synchronization of circadian clock. Furthermore, the expression patterns of clock genes in the salivary gland of larvae suggested the presence of peripheral oscillators during the developmental stages of Drosophila.

The study on circadian physiology of the high altitude Himalayan strains of Drosophila reported analysis of the effects of high altitude of origin on the circadian locomotor activity of D. helvetica. Parameters of entrainment and free-running rhythm of the haH strain of D.helvitica are unique among known Drosophila species. Its delayed but rigid phase of activity onset that refers to the lights-on transition of entraining photophase and the early but labile phase of activity termination that could be postponed by high temperature should be regarded as behavioral adaptations in response to the low temperature and other environmental conditions prevailing at the high altitude of its breeding site which have profoundly influenced the evolution of the pacemaker controlling its locomotor activity rhythm.

In vitro studies on adrenergic regulation of N-acetyl transferase (NAT) activity in fish pineal was undertaken to examine the molecular mechanism of the regulation of arylalkylamine-N-acetyl transferase (AA-NAT) activity by extensive experiments with adrenergic receptor agonists and antagonists in a teleost fish model, Clarias gariepinus. Exhaustive experimental evidences clearly indicate that (a) AA-NAT activity has a distinct circadian rhythm during all phases of the breeding cycle in the Clarias gariepinus; (b) AANAT activity is sensitive to NE at night unlike in day; and (c) AA-NAT activity is regulated by the Badrenergic signal transduction pathway (NE . B-AR . G-proteins . cAMP . PKA . PCREB . AA-NAT gene . AA-NAT m RNA . AA-NAT protein . AA-NAT activity) in the pineal gland of the Clarias gariepinus.

Herpetology

Study of distribution, ecology, reproduction and development of tree frogs of Meghalaya with special reference to Cherapunjee was undertaken to study the sacred groves as undisturbed and mined areas as disturbed eco-systems for studies. Six species of tree frogs were selected and their life history was studied taking into consideration various ecological parameters such as Ph, salinity, rainfall etc. including behavioral studies such as mating (amplexes), reproduction, parental care etc.

Vector studies

In a project on 'formulation of novel mosquito larvicide- An envisaged vector control technology'f, many soil fungi were screened, isolated and cultured, and out of fifteen fungal species, one was found to be significantly effective and useful as mosquito larvicidal candidate. A patent entitled - "A novel larvicide or insecticide has been filed. In another project, the data generated by genetic and biochemical studies of Anopheles stephensi - a malaria vector, is useful in the synthesis of transgenic strains of Anophelese stephensi which in turn could be used in the genetic control programme of the species.

Parasitology

Studies on epidemiology, intrinsic rate of natural increase and economic effects of parasitism of phthirapteran ectoparasites infesting common Indian birds, has lead to the observation of prevalence and infestation index of 24 Phirapteran species occuring on nine common indian birds. Intrinsic rate of natural increase of avian louse has been worked out. The lifetable statistics of lices were worked out and has concluded that egg morphology can be used for lice taxonomy purposes.

Biochemistry Biophysics, Microbiology and Molecular Biology

The new projects were sanctioned in the following sub disciplines:

In addition, the following inter-institutional research projects were also initiated to strengthen collaborative efforts:

Figure 2.4: SPX-40 (Signalling Protein expressed during involution/breast cancer) co-crystallized with various designed peptides

Figure 2.5: SPX-40 co-crystallized with Oligosaccharides (a) Trisaccharide, (b) Tetrasaccharide, (c) Pentasaccharide and (d) Hexasaccharide

Figure 2.6: Structure of Ternary Complexes of SPX-40 Proteins co-crystallized with both sugars and peptides together

Health Sciences

The programmes in this area were continued to be supported to strengthen the knowledge base, generate baseline data, develop newer diagnostic methods, devices, processes, and drug delivery systems, study mechanism of action, to develop work force and motivate them to engage in R&D activities in emerging and frontline areas of biomedical sciences. The sequencing of the human genomes combined with high throughput gene and protein expression technologies have revolutionized our ability to understand complex diseases in a comprehensive way. This has lead to an increasing number of challenging researches in molecular medicine and a greater demand for training in these areas. The department has been making special effort in designing training programmes/workshops for the scientific and medical professionals with special attention to latest developments in innovative frontline and emerging area of Biomedical Sciences. During this year, two workshops have been organized.

Oncology/Cancer Biology

Molecular profiling-the comprehensive analysis of genes, RNA’s and proteins- is having radical effects on our understanding of cancer. The scale of cancer research projects has increased enormous over the past decade or so, presenting scientists with the opportunity to investigate the role of human genes in cancer. During the year, several new projects were initiated in the area of cancer biology. Prevalence of hypothyroidism following treatment of head-Neck cancer and its impact on quality of life; Studies on the association of polymorphism, methylation and expression of various genes with hepatocellular carcinoma in India.

Differential expression of matrix metallo-proteins and collagen IV in oral sub mucous fibrosis; Elucidation of the role of alpha-synuclein in dopamine mediated damage to shsy5y human neuroblastoma cells; Implications for Parkinson’s disease; Assessment of the role of tyrosine kinaseEphB4 in the wnt pathways by RNA interference and its significance in colon cancer; towards novel cancer therapeutics; Development of azole derived steroidal hybrids as potent aromatase inhibitors for the management of breast cancer; Phase III trial of surgery followed by conventional radiotherapy (5 Fractions/Week) Vs. Concurrent Chemo-Radiotherapy Vs. accelerated therapy (6 Fractions/Week). In locally advanced, stage III 7 Iva, respectable, squamous cell carcinomas of oral cavity; Gene-environment interactions and susceptibility to breast cancer.

More was learnt about the genes and signaling pathways that promote the proliferation of cancer cells, prevent their death or allow their spread to other organs, to facilitate in developing new methods of diagnosis as well as treatments. Dopamine (DA) strongly and selectively inhibits vascular permeability factor/vascular endothelial growth (VPF/VEGF) - induced angiogenesis and also found to be an important regulator of IGFI (Insulin-like growth factor-I) mediated stomach tumor cell proliferation. It has been shown for the first time that there is an inverse correlation between endeavors stomach dopamine and gastric cancer. This result can be transferred from bench to clinics as an anti-angiogenic therapeutics modality of gastric cancer either alone or in combination with chemotherapy, radiotherapy or surgery. The results of the study revealed that Integrins and MMPs are important cell signaling molecules present on tumor cell surface. By blocking integrin & MMPs activity with natural products (like EGCG-Epigallo chatechin gallate extracted from tea) and curcumin, the tumor cell migration and invasion could be regulated.

The first randomized evidence suggesting that events at the time of surgery have bearing on the longterm survival of breast cancer. The hormonal milieu in the host can modulate metastatic potential of breast cancer. Studies have demonstrated significant survival benefits of progesterone treatment before surgery in patients with operable breast cancer as circulating progesterone counteract the detrimental effect of estrogen .Some new gene identified TC21/R-ras2, a small GTP binding protein, MEMD/ALCAM and G protein coupled receptor 56 in esophageal squamous cell carcinomas by differential display. These genes are associated with early stage of development of esophageal cancer. This study will be useful in designing molecular diagnostic/ pro-diagnostic strategies for esophageal cancer.

Butyric acid (BuA) has beendemonstrated as a potent antiangi-ogenic compound for the first time. BuA anti-proliferative and pro-apoptotic effect studied on endothelial cells. Curcumin ameliorates tumor-induced toxicity. Curcumin neutralizes tumor-induced oxidative stress. Curcumin replenishes tumor-induced depression of CD4+, & CD8+ T cell population. Studies shows that MRS can be used to distinguish normal, benign and malignant breast diseases and locally advanced breast cancer (LABC) patients showed that choline containing metabolites (TCho) can be used as marker of malignancy .The sensitivity of detecting TCho is 78% with a sensitivity of 86%.

To delineate the molecular mechanics behind such differential activities of curcumin, it was found that curcumin selectively increases p53 expression at G2 phase of carcinoma cells and release cytochrome c from mitochondria, which is essential requirement for apoptosis. In addition, curcumin significantly up-regulates cell cycle inhibitory protein (p21 Waf-1) in normal cells and arrest them in G0 Phase of cell cycle. These observations together suggest that curcumin may have a possible therapeutic potential in cancer patients. Polymorphism at N-Acetyl transferse 2 locus (NAT2) leads to slow, intermediate and rapid acetylating properties of the enzymes. Improper acetylation of heterocyclic and aromatic amines, present in tobacco, might cause DNA adduct formation. Generally, DNA repair enzymes removes these adduct to escape malignancy. However, tobacco users carrying susceptible NAT2 and DNA repair loci might be at risk of oral leukoplakia and cancer. Polymorphism in CYP2E1 gene might play important roles in the process of leukoplakia. Risk of cancer was also modified by GSTT1 polymorphism among “heavy” chewers and GSTM3 polymorphism among “smokers”.GSTM3 (A/A) genotype of the leukoplakia patient might become a possible marker in the process of development of cancer from leukoplakia.

Molecular iodine induces cell death in all the breast cancer cell line irrespective of their estrogen responsive and non-responsive[ER/PR status], molecular iodine activates a caspases-independent and mitochondriamediated apoptotic pathways, recent observation shows that iodine causes cell death in MDA-MB231 through autophagy and this cell death mode changes to apoptotic cell death by treatment with chloroquine an antimalarial drug.

Communicable Diseases/Parasitology/Virology

With the changes in the pattern of communicable diseases, Scientists are studying various genetic, immunological approaches to unravel the signaling mechanism, the mode of transmission, drug resistance etc.

During the year, various projects initiated included Mechanism of T cell signaling in leprosy patients by M.leprae antigen(s); Study of immune responses to common recall antigens and super antigens in autoimmune skin diseases before and after immunosuppressive intervention; In-vitro inhibition of T cell cytokine production by methotrexate: Can it predict response to methotrexate in patients with rheumatoid arthritis; Electrophysiological study of peripheral nerves in Leprosy diabetic and alcoholic polyneuropathies.

Molecular and proteomic approach for monitoring drug resistant malaria parasite in the malaria endemic zone in West Bengal; Filarial glutathione S-Transferase as a possible biochemical target for macrofilaricidal drug development – Synthesis and evaluation of substituted 1,4-Naphthoquinones. Lipid immunity to vaccine generation: identification, protective, efficacy, and mechanism of action of Leishmanial glycolipid in murine model of Visceral Leishmaniasis; Cloning and Expression of NS-1 and prM proteins for the diagnosis of dengue infections. Efficacy of some plant extracts against preadult vector mosquitoes: Isolation, characterization and field evaluation of active ingredients.

An attempt has been made to generate tetracycline regulatable null mutants of ORFF. Leishmania cells expressing the tetracycline repressor were transfected with tetracycline inducible vector expressing ORFF gene and having hygromycin, as the selectable marker. The first endogenous genomic copy of ORFF was knockout by homologous replacement with an inducible copy. Study also demonstrates that ORFF gene might be an important gene for the survival of the parasite and it may be involved in various functions like attachment of leishmania to the macrophages.

A proton –motive force (PMF) driven efflux pump which is exploited by the Vibrio cholerae to confer resistance towards fluoroquinolones have been identified and one such pump known as NorM. NorM was cloned and over expressed in E.coli. TGIAcrAB, which is sensitive to fluoroquolones due to the deletion of the gene encoding AcrAB efflux pump. Over expression of NorM in this stain, increased in MIC towards norfloxacin and ciprofloxacin specially and the increase in MIC could be correlated with the efflux activity towards these drugs. FLICE inhibitory protein (FLIP) was initially identified as a virus-encoded apoptosis inhibitory protein and its cellular homologue(c-FLIP) was subsequently found to be present in several other systems including macrophages. (c-FLIP) can be expressed either as a long(c-FLIP) or as a short(c-FLIPs) form of protein through alternative splicing. Mycobacterium avium (opportunistic pathogenic mycobacteria) induces apoptosis in macrophages. Both the FLIP constructs were able to inhibit M.Avium mediated-celldeath as measured by cell death ELISA kit. However, c-FLIP was found to be more effective. It could block cell death almost completely (>90%).It was observed that transfection with FLIP could down regulate M.aviuminduced phosphorylation of p38MAPK.

Novel HIV-I associated kinase have been discovered, which will serve as potential HIV therapeutic targets. Topoisomerase II inhibitors can form new class of HIV-I inhibitors. Mechanism action of Topoisomerase II poisoning organ metallic derivatives has been elucidated. ATopoisomerase II poisoning ferrocene derivatives has been identified as lead molecules for development of anti-HIV therapeutics. Topoisomerase beta phosphorylating kinase has been isolated to the homogeneity. Organic molecules of distinct structures shown to inhibit kinase activities, further work is in progress to synthesize new molecules. Bacillus sphaericus and Bacillus thuringenesis treatment/toxin adversely affect the development of the filarial parasite (Wauchereria boncrofti) in the Culex quinquefasciatus.The mosquito pupicidal factor (proteins) has already been purified from the culture filtrate of Pseudomonas fluorescence mw~35kDa and 55kDa. These protein were active against pupae of the three species of mosquito tested viz Culex quinequefasciautus, Anopheles Stephensi and Aedes Aegypti.

Herbal Medicine/Indigenous drugs/Drug Design

Recent years have seen considerable advances in our understanding of natural-product biosynthesis coupled with improvement in approaches for natural-product isolation, characterization and synthesis.

During the year, several new projects were initiated including Exploring the immunomodulatory potential of edible Mushroom glucan/proteoglucan as Biological Responses modifier (BRM) in cancer therapy; Phytosomes in Improving Hepatoprotective Activity of phyto-constituents; Search for Novel Bioactive Molecules as radio protectors from Ganoderma Mushrooms occurring in southern parts of India; Potential Tuberculosis Drugs from Marine Actinomycetes. Screening of herbal drugs used in treatment of hepatocellular carcinoma; A study on Antimutagenic potential of curcumin on chromosomal aberration in Allium cepa revealed that curcumin is effective at 5ìg/ml in Allium cepa root meristem cells. Study on development and evaluation of phytosomes-a value added herbal drug delivery system describes the success of phytosomes as drug delivery system may open a new avenue in the field of herbal drug research by delivering the active constituent effectively as well as improving its bioactivity. New herbal drugs – Herbal potential leads were identified in treatment of ulcer from the group of plants consisting of rhizomes of Utleria salicifolia, stem of Tinospora cardifolia ,fruits of Musa paradisca, fruits of Ficus racemosa, seeds of Maringa oleifera and leaves/fruits of Annona squamosa.

Hypoglycemic & hypolipodemic herbal drug has been formulated from the aqueous methanol mixture of the extract of seed of Tamarindus indica, which contain sugar, phenolic compound and flavinoids. Tomato in combination with garlic and turmeric demonstrated anti-genotoxic/ant carcinogenic potential by decreasing lipid peroxidation and enhancing the antioxidant status by decreasing Bcl-2/ Bax ratio and upregulation of Bim, P53 and capsases. Tomato and garlic can act synergistically to inhibit HBP carcinogenesis. New Arecoline derivatives have been synthesized to investigate their affinity and potency towards M1 receptor in relation to learning and memory for the applicability of these molecules in Alzheimer’s dementia.

Diagnostic Methods

A method to label a cohort of erythrocytes that is made and released in blood stream during a defined window of time and then track age related change as this erythrocytes cohort ages in blood have been devised. A simple, rapid, cost effective sensitive, reproducible laboratory method for the early diagnosis of tuberculous meningitis (TBM) by the direct immuno-cytochemical methods for the demonstration of microbacterial antigen in the cytospin smear of CSF specimen has been evaluated.

A blood-based assay for detection of acute lymphocytic leukemia (ALL) developed and patented. This is based on expression of 9-O-AcSGs (O-acetylated sialoglycoconjugate) on cell surface of erythrocytes 9-OAcSGs. These sialoglycoconjugates induced on erythrocytes can also serve as diseases specific biomarkers. O-acetylated siologlycoconjugate have shown altered expression in various patho-physiological conditions. 9-O-AcSa Derivatives are selectively present on leukemic blast of children with ALL. 9-O-ACS is total absent in normal erythrocytes. These sialoglycoconjugates induced on erythrocytes can also serve as diseases
specific biomarkers

Life style Disorder: Diabetes/ Ocular diseases/Neurological disorders/Respiratory disorders

Life style disorder has genetic predisposition. Emerging genome resources and technologies are enabling system identification of genes underlying these complex traits.

With the increasing trend in life style disease, various new projects were initiated which included Biochemical and Molecular Basis of Type-II Diabetes-induced Cataract: Evaluation of a Suitable Animal Model and Role of Dietary Agents; Molecular Pathology of Hypertrophic Cardiomyopathy; Study on Mutations of Genes of Monogenic Forms of Diabetes in the Young-Onset Type 2 Diabetes mellitus in Eastern India; Regulation of endothelial barrier function by chronic hyperinsulinemia and lipotoxicity: Implications in diabetic vasculopathy; Silencing of ER stress response genes using RNA interference to protect hyperglycemia induced pancreatic B-cell death; To bioengineer a corneal stromal substitute using crosslinked connective tissue; Molecular analysis of the retinoblastoma (RB1) tumor suppressor gene product in human neurofibromatosis type 2 (NF2) and related intracranial tumors; Gene expression profiling and development of microarray-based biomarker for diabetic retinopathy; Assessment and validation of Body composition using different techniques and development of regression models in Indian Population; Early detection of polarized cytokine producing allo-reactive T cells and the role of chemokines in their selective homing among patients undergoing renal transplantation; Non-linear dynamics and time series analysis of respiratory rhythms; Study of genetic polymorphism of dihydropteroate synthase (DHPS) gene in pneumocytis jiroveci isolates and its correlation with treatment failure & clinical outcomes in patient with pneumocytis carinii pneumonia; Validation of the Physiological Cost Index by comparison to oxygen consumption and it is potential as an outcome measure of ambulation in children with cerebral palsy; To investigate the effects of maternal hypothyroidism on transcription factor sequel in early neurogenesis and neuronal migration; the effects of continuous positive airways pressure therapy for prevention of new vascular events in patients with sleep disordered breathing and stroke: A randomized controlled study.

CYP2C enzymes of cytochrome P450 group plays an important role in drug metabolism and CYP2C8,CYP2C9 and CYP2J2 genes are shown to be highly polymorphic and frequency of allelic variants differ based on ethnicity. Studies are under-progress to delineate association between CYP2C8, CYP2C9, CYP2J2, and eNOS gene polymorphism and susceptibility to acute myocardial infarction in the south Indian population. Agmatine induced eNOS phosphorylation is reported for the first time .Presence of nitric oxide synthase in the nucleus and different intracellular organelles ha also been reported for the first time in neutrophils. Anti-diabetic activity of various fraction of Enicostemma litorale (Whole Plant) has been established using various animal models and “Swertiamarin” an active compound have been isolated. A novel process of isolation of swertiamarin compound from plant Enicostemma Litorale has been patented.

Results provide strong support to the contention that Diabetic Retinopathy is an inflammatory disease. On the basis of analysis of well-defined homogenous set of patients, inverse relationship was observed between VEGF and PEDF, indicating the importance of imbalance between these angiogenic and antiangiogenic factors in pathogenic neovascularization. Studies are under progress to modulate MMPs activity, in the environment of the ECm of proliferating retinal cells through addition of MMPS, as on alternative to releasing MMPs from fibronectin-bound states. This will help us to establish whether higher availability of mmps/collagenolytic proteases disrupt the ECM surrounding proliferative retinal cells to reduce membrane formation through ECM breakdown or facilitative ECM remodeling.

Results of the study on Anthropometric, Biochemical and genetic contribution to Visceral and subcutaneous adiposity in diabetic and Non-diabetic subject’s shows that central abdominal fat measured by DEXA is also observed to be higher in Diabetic subjects. The observation that only younger diabetic subjects had significantly higher visceral and central abdominal fats suggest that, with age, even non-diabetic subjects accumulate some visceral adiposity and in younger individuals, visceral adiposity probably predisposes to diabetes. Some dietary agents like curcumin (as inhibitors of aldose reductase and proteinglycation) have modulating role on á-crystalline chaperone- like activity, which may be exploited to prevent or arrest the progression of diabetic cataract.

Three SNPs in GRIK1/GluR5 gene in control and parental samples have been characterized and their magnitude significance of linkage disequilibrium between these loci has been determined for studying polymorphism that confers susceptibility to mental retardation in Down syndrome.

Further study determines that the 522(A/C) and 1173(C/T) markers may be informative in our population samples as the alleles frequencies are significantly different from those reported in other asian population samples. Bio-informatics analyses indicate that alternative splicing of the primary GRIK1/GluR5 transcript generates two distinct mRNA species In Eale’s Diseases, involving retinal changes such as perivascular phlebitis followed by neovascularization. the study demonstrate for the first time the presence of increased levels of VEGF in vitreous. It is significant that in this idiopathic inflammatory condition, neovascularization is also mediated by VEGF.

Genetic Disorders

Genetic screenings are powerful tools for identifying the gene involved in specific biological processes. Gene expression profiling is already being introduced clinical as a diagnostic procedure. During the year several new project such as Lupus Nephritis: Studies on Immunogenetic profile of Patients from Southern India; Genetic Polymorphism in Indian Thyroid cancer patients; Genomic instability in Bystander Human Cells and their Progeny exposed to Radiomimetic Drug using 3-dimensional cultures; Typing of Pseudomonas aeruginosa in cystic fibrosis patients in Northern India; A study on prognostic impact of molecular genetic alterations in Hepatoblastoma in Asian Indian Children; Molecular study of fragile X Syndrome in UP Population; Direct mutation analysis of FVIII gene in Indian Hemophiliacs; and Genetic analysis of apoptotic pathways involved in macrophases from bruton’s tyrosine kinase-deficient X linked immuno-deficient mice were initiated.

It has been documented for the first time, that the disease causing mutations of ATP7B gene in Wilson’s diseases patient, predominantly from south India.11 novel mutations were identified and shown to have pathological significance in causing Wilson’s disease in our population and these mutations have been indexed the human genetics mutation database as “Novel pathological mutations”.

Studies have shown strong over representation of alleles in kidneys that work initially normal but progressively exhibited renal dysfunction. The I/D polymorphism of the ACE gene has shown to modulate circulating and tissue activity of angiotensin converting enzymes, that is individuals with DD genotype have the highest serum ACE levels, those with II genotype have the lowest and those with DI genotype have intermediate levels. The D alleles may be one of the genetic susceptibility factors contributing to adverse renal prognosis and patients with congenital uropathies, especially if the anomaly is Vesico-ureteric reflux or posterior urethral valves The potential usefulness of genotype evaluation extends beyond prognostication of parenchymal damage, to identification of patients who may benefits from angiotensin converting enzymes inhibitors and angiotensin II antagonist.

Reproductive Biology

To meet challenges posed by changing health scenario and incidence of infertility and to improve reproductive health several new projects including Cloning, expression and functional characterization of a cyclin-like protein (CLP-1) from human germ cells; Elucidating the Role of Progesterone in Cervical Ripening using Rat Model; Testicular germ cell apoptosis in diabetic albino rat: Corrective effect of a formulated herbal drug – MTEC and its clinical trial; Modulation of neural-immune signaling by estrogen in reproductive aging; An investigation on the molecular basis of male infertility in tribal people of north eastern India; Proteomic approach for mapping and characterizing domain specific epididymal proteins involved in post testicular sperm maturation were initiated.

A sensitive way for superoxide detection and estimation in the spermatozoa was established. A molecule designated as Cyclin-like protein-1(CLP-1) expressed differentially in fertile males, which play distinct role in cell cycle regulation was identified. In an ongoing project on defining endometrial receptivity for blastocyst implantation using cDNA based array technology, a set of genes have been identified that expressed deferentially in monkey endometrium and human endometrium epithelial cells under progesterone dominance as compared to control and antiprogestrone treated tissues. Further studies are needed to achieve space and time contexts of functional clusters of transcripts. Validated and optimized the cDNA based transcript array technology. The protocol for designing primers and probes for performing real time PCR in multiplex format using the Beacon Designer software obtained for the present project was standardized.

Considering the fact, that sperm maturation is a prerequisite before it can fertilize the egg many groups are interested in understanding the basis of sperm maturation. One approach is to identify the sperm specific proteins and attempts to understand their role. A step towards this is the establishment of the proteome of the

sperm and to identify domains specific epididymal proteins by subjecting the epididymal sperms from different regions two classical 2D Maldi TOF and compare with testicular sperms. Following, this the gene for identified specific proteins will be cloned and recombinant proteins will be used to ascertains the function Studies are under progress on development of newer contraceptive approach as well as identifying the cause for infertility.

Occupational Hazards

In a project, for physiological and metabolic studies on cycle rickshaw pullers before and after strenuous exercise an Instrument has been designed for cycle rickshaw pullers for assessing precision grip force using a battery-powered device. This will be used on patients to quantify their recovery after intervention. This can assess sensory, motor and their integration with respect to hand function. The virtual loading methods can be used to develop a model for system behavior.

Stem Cells

Studies are under progress for developing method for creating niche for expansion of long-term multilineage engraftable hematopoietic cells. New and indigenous methods have been developed for studying the age related changes in erythrocytes in blood circulation. Technology for establishment of hES cell lines derivation in a xeno-free environment using human feeders is being developed. 4 cells lines have been developed from blastocyst and are in early stage of cultures.

Plant Sciences

Some of the projects funded during the year were:

Fine mapping of fertility restorer genes for WA-CMS system of rice and its application for identification of restorers; Iron metabolism in rice (Oryza sativa L) plant with emphasis on its translocation and assimilation; Identification and cloning of miRNA that are dis-regulated under biotic and abiotic stress in rice; Transfer of traits for salinity tolerance from Leptochloa fusca (kallar grass) to Oryza sativa (rice)-A non conventional strategy; Oxidative stress and inducible tolerance mechanisms in rice plants under aluminium toxicity; Metabolic engineering of gibberellins signal transduction pathway for increasing the yield potential of indigenous aromatic rice cultivar; Role of serpin, a conserved protease inhibitor, in regulating rice disease defense responses; Decaffeinated Tea: Through gene silencing; Isolation and characterization of heterotrimeric G-proteins (subunits: alpha, beta and gamma) and G-protein coupled receptor (GPCR) from Pisum sativum: study of their regulation under stress; Genetic transformation of Ginger to improve fungal resistance against Pythium aphanidermatum; A proteomic approach to identify the proteins involved in in vitro differentiation of Vigna radiata; Isolation and characterization of starch and protein from different legumes; Cloning, characterization and expression of drought stress responsive genes from horsegram (Macrotyloma uniflorum Lam. Verdc.), a dry land legume grain crop.; Identification and regulation of the key gene involved in bixin bixin biosynthetic pathway through differential expression analysis in Bixa orellana L.; Physiological responses of crop plants to water deficit under elevated CO2; Molecular Characterization and Development of diagnostics for Virus(es) and Viroids Infecting Apples; Molecular characterization of Pongamia pinnata germplasm; Screening of molecules of plant origin as inhibitors of solid surface colonization by Candida albicans; Targeting of the bioactive molecule hapalindole-T isolated from cyanobacterium Fisherella sp etc.

The important results of the ongoing projects are given below:

In the project “Screening of Aloe vera L. germplasms for cosmetic gel and micro-propagation of elite clones” the Germplasms of Aloe vera L. collected from different agro-climatic zones of India were screened for cosmetic properties in relation to UV- opacity, antioxidant, antiobesity, tyrosinase, and immuno-modulatory activities along with physical characterization of the gel. Among the germplasms, there were considerable variations in cosmetic attributes. UV-opacity potential was documented for the first time in aloe gel with a range of sun protection factor (SPF) 1.88- 3.49 among the germplasms (Figure 2.7typical peaks of DMPO-OH adduct formation (Figure 2.7 b) and presence of selenium suggests the antioxidant potential. Aloe gel had potent inhibitory effect on mushroom tyrosinase. Lineweaver-Burk plots showed competitive inhibition. Inhibition of pancreatic lipase both by colorimetric and spectrofluorometric assays was evident with aloe gel. Aloe gel also reduced the body weight, adipose tissue weight, liver weight, and cholesterol and triglycerides of mice fed with high fat diet. The findings suggest the antiobesity effect of gel. a). EPR signals with


Figure 2.7 a) Absorption coefficient of lyophilized gel (4 mg/ml) of Aloe germplasms b) EPR spectra showing scavenging activity against hydroxyl radical from water extract of aloe gel (germplasms S24) at 1mg / ml concentration.

Figure 2.8: The zbf1 Mutants Show Multiple Phenotypes. In each panel segregated wild type (Col) and zbf1(atmyc2) mutants are shown on the left and right side, respectively. A: Six-day-old constant dark grown seedlings. B: Six-day-old constant RL (95 mole/sec/m2) grown seedlings. C: Six-day-old constant BL (30 mole/sec/m2) grown seedlings. The zbf1 mutants have shorter hypocotyls. D: The root growth of 16-day-old wild type and atmyc2-3 mutant plants. The zbf1 mutant plants have lees lateral roots formed compared to wild type

In the project on “Investigating the molecular mechanism of auxin dependent stress tolerance in plants using an Arachis hypogea cell suspension culture” it was investigated that ABA responsive transcription factor ABI3 (ABA insensitive 3) was differentially expressed in presence of auxin in an auxin autotrophic stressed Arachis cell culture. It was also demonstrated that by virtue of having a DNA binding domain that is similar to ARFs (auxin response factors), ABI3 can bind Auxin responsive elements (AuxRE). Binding of ABI3s and ARFs with the AuxRE was mutually exclusive (Figure 2.9).