Annual Report 2006-2007

Life Sciences

Animal Sciences

The Programme for Animal Sciences aims to promote basic science research in the areas. During the year under report activities aiming at generating quality research manpower through SERC Schools and training-cum-brain storming sessions were organized. New research projects were sanctioned on subdisciplines such as taxonomy/biodiversity, ecology, behavior, chronobiology, toxicology, endocrinology and physiology and developmental embryobiology. A total of 20 projects were sanctioned at a cost of Rs.3.58 crores for 3 years and Rs.2.00 crores were released with a lion share of Rs.1.62 crores towards University sector. A brainstorming session-cum-conference was organized at Zoology Department of Lucknow University on, "Helminth taxonomy - revisited" to find out ways and means to strengthen the academic activities further.

Soil Biology: Heavy application of pesticides and fertilizers has resulted in degradation of soil productivity. There are various interfaces from different angles to restore soil health. A project to assess the suitability of fly ash as soil amendment with special emphasis on toxicity to earthworms and microflora has been sanctioned at Department of Chemistry, Pune University. Another proposal to evolve biomarkers of 16s rDNA, enzymes, microbes and animals as model systems has been sanctioned at Zoology Department of Delhi University.

Soil health is a major concern and earth worms play very important role in maintaining it. A project on earth worm resource and their utilization was sanctioned to study in rubber plantation at a college in Tripura. Water is an integral part of soil. A proposal to study synthesis, cloning and expression of organophosphorus hydrolase for molecular bioremediation of insecticide contaminated drinking water has been sanctioned. Division of Soil Science & Agricultural Chemistry, IARI, New Delhi was awarded research project to probe microbial diversity in relation to soil health and productivity of the Central Himalayan region.

Faunal Diversity: A proposal to study biodiversity of Pulicat lake sanctuary has been sanctioned to Sri Venkateswara University, Tirupati. Another proposal on amphibian and reptiles diversity has been sanctioned to study their adaptation and ecology in Northern part of Rajasthan at MDS University, Ajmer.

Behavior: The Centre for Ecological Sciences, IISc, Bangalore has been sanctioned a project to study acoustic communication in Asian elephants and its correlation with behavior. A similar study for optimal foraging behaviors of the Indian false vampire bat Megaderma lyra has been sanctioned at MK University, Madurai. Chemicals from the plant influence behavior of its pests and a project to study influence of plant chemicals and physical characteristics of Acheae janata on castor semilooper has been sanctioned to IICB, Hyderabad.

Chronobiology: Chronobiology is an area which is comparatively new but fairly vibrant in the country right now. The Animal Science programme has launched a multiprong programme to serve the area. A project to study circadian pattern of expression of dclock and vrille in brain and peripheral tissues of cryb and vg mutants of Drosophila has been sanctioned to Annamalai University. An IRHPA Unit, has been sanctioned at the Lucknow University to study rhythmic behavior of birds from molecule to behavior at a cost of Rs.2.50 crores.

Physiology: Research proposals related to physiology and endocrinology have also found proper representation. Research projects to study role of selected plant hormones on animal cell function, physiological significance of expression of different types of nitric oxide syntheses and production of nitric oxide under environmental constraints and pathological conditions in catfish; influence of stress and stress related peptides on ovarian functions in the fish; functional analysis of ecdysone regulated ecdysone receptor etc have been sanctioned.

Developmental Biology: Developmental Biology is emerging as an active area. A project on biology and embryonic development in Cynopterus sphinx in respect of various metabolic factors, growth factors, follicular growth and atresia has been sanctioned to study goat ovaries at Department of Zoology of Kurukshetra University; and heme oxygenase 1 in rodent embryogenesis has been sanctioned to the Department of Zoology, Delhi University.

Achievements

The research projects on mammalian ecology have been implemented and now majority of them are complete. The salient achievements are listed below;

Parental investment: The project on maternal parental investment patterns in relation to dominance, rank and sex among the lion-tailed macaques have come out with interesting findings. The pattern of investment indicated that the male lion-tailed macaques garner more maternal investment than females. However, the dominant females largely invested more on sons and subordinate females on daughters. Since infants inherit mothers rank socially, a son of a dominant female is likely to become dominant and produce larger number of offspring. A subordinate female's daughter may remain subordinate but all females produce with little variance in reproductive success. The parental investment patterns, therefore, are not random but products of natural selection as these are strategies of parents to maximize their own inclusive fitness.

Reproduction: The project on development of fecal steroid analysis as a non invasive method to monitor reproductive function in Indian endangered animals (big cats and ungulates) aims for successful captive breeding. Basic understanding of the reproductive potential such as fertility status, time of ovulation, optimal time for mating, pregnancy, time of spermatogenesis etc. of the female is necessary. The studies were undertaken to identify major fecal steroid metabolites so as to develop antibodies against the major metabolites and to develop a radioimmunoassay for the Indian endangered animals. Preliminary studies have been conducted to identify estrogen metabolites in fecal samples of spotted deer, swamp deer, hog deer and barking deer. The most predominant metabolites in all the deer species were estradiol sulphate (E2SO4), estrone glucuronide (E1-G), estrone sulphate (E1-SO4) and estriol (E3). Further studies are in progress to identify these metabolite in big cats such as lion and tigers and progesterone metabolites in ungulates.

Communication : Communication among animals is very interesting area of study. The project on localization of prey by the Indian false vampire bat Megaderma lyra has progressed well. Studies show that the M. lyra in addition to passive listening, uses its echolocatory system also to detect prey on the ground. While pulling a freshly killed frog, the bat approaches it by emitting high frequency echolocation sounds containing 4 to 6 harmonics (peak at 45 kHz).

Conservation : There are two projects on ecology, status and conservation of lesser known mammals in neglected habitats throughout the state of Karnataka covering a few protected forests and all reserved forests and private lands and the other one on ecology and conservation of leopard in Gir National Park and Sanctuary, have been implemented. The ecological correlates and conservation status of wild mammals except burrowing rodents and bats was assessed taking each taluk as the basic unit for presence/absence/ density of a species. The pattern on human animal interaction has been illustrated and few flagship species of mammalian conservation in Karnataka were identified. These species are lion-tailed macaques for the rainforests, tiger and elephants for deciduous forests; slender loris for dry and scrub forests and wolf for the plains.

Slender Loris being studied for mamakian conservation in Karnataka.

Based on the data provided out of this project, strategic regions in Tamil Nadu were identified for loris conservation and all the data has been provided to Karnataka Forest Department to plan for conservation and management of mammals.

The leopard project in Gir National Park investigated aspects such as habitat use, ranging and movement pattern, seasonal food habits, social organization and leopard human conflict in and around Gir protected area. Four leopards and one leopardess were captured and fitted with radio collars.

A total of 318 kills of leopard were located during the study period from 2002 to 2005. 11 species were found killed by the leopard during the study period. Two species, chital (61.32%) and peafowl (29.56%), comprise the 90% diet of the leopard. The niche overlap in terms of food items of leopard with that of lion was 67% during the study period based on 1000 simulations performed in EcoSim.

Another project to study ecology and conservation of Indian wolf in some selected habitats in Maharashtra was implemented. Wolves from four packs were radio collared for collecting data and food habit, predatorprey relationship, i.e. wolf blackbuck interactions, ranging patterns, reproductive biology and conservation and management of the species. Analysis of 3947 scats collected from 14 wolf packs revealed 27 food items

Leopard being filted with nadio collar in Gir National Park

Territorial Male Blackbuck killed by wolues

in their dietary spectrum. There was prey switching between different food types consumed by wolves. Domestic livestock comprised 47.16% followed by wild prey 38% whereas vegetable matter 14.6% and some unidentified food material. Two of the four radio collared wolf packs that were monitored extensively for their movement patterns ranged from 213 to 229 sq.km. area whereas the other two that were monitored less extensively ranges from 129 to 163 sq.km. area. A sub-adult male and the same age group female during dispersal from their parental packs ranged within 724 and 480 sq.km area respectively before they settled in new areas and established new packs there.

Instrument : An equipment Electronic micromotility meter has been fabricated and submitted for patenting out of one project being implemented at a college. It has potential for commercial exploitation. The specific uses of the product will be;

• To record the motility of the microscopic larval nematodes, trematodes and the adult parasites.
  
• Quantitative measure of percentage reduction in the motility of drug-treated parasites.
  

Policy Initiatives : New guidelines for wildlife research by Ministry of Environment and Forest has been finalized and many enabling clauses for scientists were inserted in the documents, based on the inputs provided by Department of Science & Technology.

The input for Animal Sciences towards XIth Five Year Plan were provided with many new initiative such as freelance scientific workers, DST faculties at Universities and setting up institutions in identified areas etc.

Health Sciences

The Department continued to support Bio-Medical Sciences to strengthen the knowledge, 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 R&D activities in emerging and frontline areas of biomedical sciences. 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 frontline and emerging area of Biomedical Sciences.

Over the last decade in particular major advances have been made in our under standing of both normal biological process and molecular mechanism underlying a variety of medical diseases. Knowledge obtained from basic cellular molecular and genetic studies has enabled the development of strategies for the modification, prevention and potential cure of human diseases. To integrate research in molecular biology into the clinical setting, a centre of excellence in the field of molecular oncology being established .in the southern part of the country. The centre proposed to study some of the common cancer in southern India such as breast cancer, cervical cancer and gastric cancer. Gene expression studies using microarray will be done to identify specific gene/pathways involved in the maintenance of the malignant phenotype.

An advanced research programme has been initiated to develop and strengthen the research on neurotransmitters such as Acetylcholine, Dopamine, Epinephrine, Norepinepherine, Serotonin, Glutamate, GABA and their receptor subtypes and associated disorder by studying the molecular mechanisms of diseases and their management.

With the increasing incidences of cancer in the country, several new projects sanctioned to individual scientists to study the various aspects of different types of cancer e.g., role of Estrogen and 17 Beta Hydroxy Steroid dehydrogenases as tissue specific factors in the etiology of BRCA1 associated Breast Cancer; Antisense technology based development of stable Insulin-like growth factor II (IGF II) phosphorothioate antisense oligomers and study of their efficacy as a future drug for liver cancer; Evaluation of therapeutic effect of Dopamine on human stomach tumor growth in nude mice; Molecular markers of oral submucous fibrosis by gene expression profiling; Real time PCR quantitation of minimal residual disease (MRD) and detection of SNPs in drug metabolizing enzymes in ALL patients.

There is increasing trend observed in various life style diseases like Cardiovascular disorder, Genetic disorder, Diabetes, Renal Disorder, Respiratory diseases, Reproductive Biology, Gastroenterology, Neurological disorder and Hematological disorder. The Department has sanctioned various projects in these areas e.g., evaluation of the Mechanism of the Synergism between Purinergic and Adrenergic receptors and search for Platelet based markers of ACS (Acute Coronary Syndrome); Intravascular Ultrasound for Severe Pulmonary Artery Hypertension in Congenital Heart Diseases; Studies on mechanisms operative in development of diabetic cataract; Biomolecules from Tamarind indicus and Terminalia arjuna influencing mineralization I demineralization processes and their role in the management of Renal Calculosis; Pathogenesis of nonsteroidal anti-inflammatory drug (NSAID) induced renal damage; Investigations on artificially produced Urinary Stones composed of natural materials; A study of methods for reducing exert ional breathlessness and increasing exercise capability; Assessment of nutritional and antioxidant enzyme status in mechanically ventilated critically ill patients. Effects on mortality and strategies for improving nutritional support; Effect of capsaicin on endothelial cell function and angiogenesis; Structural and Molecular Signatures of Early Pregnancy on Primate Endometrium; Defining endometrial receptivity for blastocyst implantation using cDNA based array technology; A Genetic Analysis of Polycystic Ovary Syndrome with Special Emphasis on Genes involved in Insulin Resistance; An Evaluation of Genetic and Molecular Factors contributing to premature ovarian failure in Indian population; The effect of chronic immobilization stress on electrophysiological properties of the hippocampus and amygdala. Probing the GRIKI / GluR5 sequence for polymorphisms that confer susceptibility to mental retardation in Down syndrome; Gene mapping for Myopia and identification of environmental risk factors for myopia progression in subjects with genetic susceptibility; Investigating the role of BLM helicase and signal transducer, 53BP1 during homologous recombination in human

Vector born diseases, viz., Malaria, Filariasis, Kala-azar, Dengue and Japanese Encephalitis (JE) are major public health concerns and impede socio-economic development. The Department has sanctioned several projects in the area of Communicable and Vector Born Diseases to study indepth the vector biology, genomics and proteomics of the causative organisms e.g., Molecular Mechanism involved in M. tuberculosis entry/survival within human macrophages; Molecular analysis of Environmental Vibrio cholerae Strains: Relationships with clinical isolates; Toll Like Receptor 2 (TLR 2) Polymorphism in patients with Mycobacterial diseases; Role of Interferon-gamma and Interleukin-12 transformed Dendritic cells in preventing and / or clearing Mycobacterium tuberculosis infection; Pathogenesis of tropical sprue : Study of antroduodenal manometry duodenocecal transit time, fat-induced ileal brake and small intestinal permeability; Development of molecular diagnostic assays for members of the Anopheles culicifacies sensu lato complex for improved malaria vector surveillance; Development of molecular and genetic markers of virus transmission competence of dengue vector species in Rajasthan; Prevalence of Wolbachia infection in the filariasis vector Culex quinquefasciatus in Villupuram District; Evaluation of Innate and Adaptive Immune response of endemic normal and Asymptomatic Microfilaremic individuals to recombinant parasite antigens in lymphatic filariasis; Identification, characterization and role of proteinases during the pathogenesis of Leptospirosis; Isolation and characterization of Proteophos- phoglycans of Leishmania donovani;

Drug discovery and development of new drug molecules needs the study of what the drug does on the body (Pharmacodynamics), curing the disease without causing harmful side effects, and of what the body does on the drug (Pharmacokinetics), namely ADME absorption, distribution, metabolism and excretion of the drug. The department has sanctioned several projects to explore activity of indigenous medicinal plants and studying activity of the various drugs e.g., antioxidant Phyto-chemicals in Edible Plants and their effects on Lipid Oxidation in Food and Biological systems; Characterization of immunotherapeutic potential of extracts prepared from native and in-vitro cultivated high altitude antimicrobial plants; Evaluation of the chemopreventive potential of neem (Azadirachta indica) leaf fractions in the hamster buccal pouch carcinogenesis model; Studies on the aphrodisiac properties of Vanda tessellate flower extract for determining its utility as a commercially viable standardized phytomedicine; Discovery of Potential Antileishmanial Chemo-therapeutics and Ethnotherapeutics from Medicinal Plants; Phytotherapy as a Remedy for Calcium oxalate induced Kidney Stone - A Biochemical study in Rat Urolithiasis model.

Results of the studies on some of ongoing projects have reported vital information related to some of the genetic diseases such as Alzheimer's diseases, Cystic fibrosis etc. In a study on expression and regulation of PS gene involved in Alzheimer's disease and its regulation by estrogen hormone in aging mice brain revealed that estrogen and testosterone play important role in PS1 and PS2 gene expression. The level of PS protein decreases with age and is regulated by estrogen and testosterone.

Cystic fibrosis is a hereditary disease caused by defected gene and is an autosomal recessive disorder. Results of studies on identification & characterization of mutant Cystic fibrosis transmembrane conductor gene (CFTR) suggested that delta F 508 mutation was detected in 13 out of 64 chromosomes studied. A novel mutation lle in exon11 of two patients identified. A study on differential expression of gene of esophageal cancer yielded interesting results. Recently some new gene identified TC 21 / 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, probably mediating cell -cell interaction promoting cell clustering. This study will be useful in designing molecular diagnostic / pro-diagnostic strategies for esophageal cancer, based on the validation of these candidate molecular markers.

von Willebrand disease ( vWD) is another autosomal inherited bleeding disorder. The genome for vWD is located at chromosome 12p 13.2. Studies are under progress to under stand the underlying pathogenesis of the von Willebrand disease using molecular techniques and are likely to elucidate the optimal therapy for common variants. To understand the transcriptional regulation of thyroid hormone (T3) induced cardiac hypertrophy, cDNA microarray analysis of heart tissue were performed. The T3 altered gene encodes various types of proteins related to metabolism, matrix and cytoskeletal structure, growth factors, transcriptional receptors, Calcium channel etc. Physiological significance of one of such altered protein, insulin-responsive glucose transporter type 4 (GLUT 4) and annexin A6 in hyperthyroid heart was studied in details. The expression of GLUT 4 was drastically reduced in the ventricular tissue of hyperthyroid heart. Insulin - induced glucose uptake in hyperthyroid cardiomyocetes was reduced significantly. Annexin A6 is upregulated in hyperthyroid rat heart and is localized to the Ca2+ release channel in cardiomyocyetes and functionally coupled to the enhanced Ca2+ oscillation by thyroid hormone (T3 ).

Several projects have been implemented to develop new diagnostic methods, devices, drug delivery system as well as synthesis of new compound as new drug molecule. In the field of pharmaceutical technology, there is an urgent need to search for non- parental route of administration of potentially active peptides / proteins and some synthetic drugs because of their degradation by gastric acid and extensive first pass metabolism by liver. In an attempt to deliver the drug through transmucosal route, novel muco-adhesive buccal tablets along with penetration enhancers were developed to deliver the drug unidirectional and directly to the systemic circulation. Natural mucoadhesive substances were extracted from plant sources (fruits, seeds and leaves)

For the first time an attempt has been made to study relationship between delusional traits and HLA gene. Data obtained so far implicates HLA -A *03 gene is associated with the disease significantly and leads to our understanding of pathophysiologic mechanism of this complex diseases. So HLA*03 gene may be used as a biological marker of delusional disorder. Developments of new non-invasive diagnostic methods are very important to validate existing invasive diagnostic methods. Result of the studies on evaluation of the role of MRI and MR spectroscopy in non-invasive detection, assessment of treatment response and tumour metabolism in breast cancer indicate that MRS methodology can be integrated with routine MRI investigation to aide further diagnosis and assessment of tumor response to therapy . In as another study a prospective randomized controlled trail assessing the impact of primary progesterone therapy in women with operable breast cancer being conducted, to evaluate the effect of pharmacological level of progesterone at the time of surgery on survival in women with operable breast cancer. A meta-analysis suggested 15 % difference in favour of women who had surgery in luteal face, which was further confirmed by circulating progesterone levels at the time of surgery showing a protective effect against mortality.

Development of bioactive polymer composite material and characterize it for its suitability as a bone graft substitute in spinal fusion surgery is a recently evolved concept. Result of the study on development of biocomposite polymaterial showed excellent bioactivity for the composite system that contains 30 and 35 vol percentage bioactive glass in the polymer matrix. Pre-drying of the PBT matrix is found to be very important to avoid degradation of mechanical properties of composite produced. Since the composite material is, promoting osteoblast cell proliferation it may be assumed that composite would be bioactive and promote bone growth on implantation. Composite exhibits adequate mechanical properties too and has potential to be used for the reconstruction of bones in maxillofacial, cranial, middle ear, vertebral areas. It may be used for replacing bones after multiple bone fracture, tumour removal etc. Composite can also be used as screw for bone fracture fixation. Since the screws are expected to be bioactive and non-degradable, there is no need of removing the new screw after fixation.

In a project on process optimization and development of biodegradable and disposable poly (propylene fumarate) polymer based bone cement for the minimally invasive radiological procedure for treatment of the bone diseases, new procedure has been followed for the synthesis of injectable and biodegradable poly (propylene fumarate) resin. Optimization for the degradation has been carried out by controlling the crosslink density of cured poly (propylene fumarate) polymer. The degradation and release of acidic components was also optimized to control the pH required or the biosafety of this cement. Studies are under progress to find out inflammatory cells around implants to predict chronic immune response to biomaterials.

Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation. It occurs in about 1% of the population and is characterized by destruction of cartilage and bone. The presently available disease modifying anti-rheumatic drug (DMARD) have their limitation in that all of them are effective 50 to 65 % of patients. To understand the mechanism of action of the DMARD and designing therapies that is more effective, the effect of in-vitro treatment of fibroblast-like synoviocyetes (FLS) with sulphasalazine (SSZ), methatrexate (MTX) and gold sodium thiomalate (GSTM) on the capacity of IL-1â stimulated FLS to produce pro - inflammatory cytokines , metalloprotease 1 and 3 and their inhibitor TIMP have been evaluated. While STM were able to suppress IL -6 and IL -8 productions, MTX was only able to suppress IL -6 and not IL -8 levels. None of the drugs had any effect on IL 10 levels. All the three drugs could suppress MMP -1 and MMP -3 productions with no effect on TIMP levels. The effect of SSZ was many fold more than of GSTM in all instances.

Multiple antibiotics resistance has emerged as a problem in the treatment of infections caused by Gramnegative pathogens like Vibrio cholerae, the causative agent of cholera. Fluroquinolones normally possess excellent activity against V. cholerae. Studies are under progress to understand the mechanism of drug resistance in Vibrio cholerae. A detailed mutational and topological analysis of N or M of V.chloerae with particular aim of identifying residues crucial for imparting FQ resistance. Results demonstrate that the important amino acid residues of E124, G184, K185, G187, P189, E190 on periplasmic loops, D155 and Y384 on cytoplasmic loops and C196 in a transmembrane segment in the NFX resistance imparting property of N or M. N or M was expressed in E.coli to evaluate the contribution of N or M to drug resistance.

Beneficial influences of dietary garlic and onion on the induction of cholesterol gallstone disease was evaluated in experimental mice. Dietary garlic and onion countered hyper cholesterolemia 29-34 %, by the decreasing LDL-cholesterol by 41-49 %.The decrease in phospholipids content in serum brought about by lithogenic diet was also significantly countered by dietary garlic and onion , dietary fenugreek seeds significantly lowered the incidence of cholesterol gallstone in mice .The drug of choice at present for the control of lymphatic filariasis is DEC (Diethyl carbamazine ) citrate, which is only microfilaricidal and its action on adult worms is doubtful. The procedure for synthesizing (1-N-Methyl-4(substituted)benzoyl piperazides has been standardized. From the study so far out of 26 compound, six compound DSTD11,12,13,14,15,and 24 were found to be promising for macrofilaricidal activity in the screening against Setaria digitata worms under in-vitro condition. Antibody against HEV replicase was generated in the rabbit, which are used for replicase immunoflorescence and immunoprecipitation analysis.

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. Retinal endothelial cell culture system has been established by isolating the primary cells from cadaveric eye of human donors. The HREC system appears as a unique cell model to test the effect of high glucose on various molecular processes that regulate cell proliferation such as apoptosis, oxidative stress and VEGF expression and the substances that inhibit proliferation. Monensin is very effective in treatment of chloroquine- resistance Plasmodium faliciparum malaria. It has been demonstrated for the first time that liposome can be used for the delivery vehicle for monensin in vivo. In a study on diagnosis of myocardial cell damage by novel radiotracer have demonstrated that 99m Tc-cysteine exhibits much better uptake than 99m Tc- glucaric acid in globally damaged heart model. The uptake of 99m Tc-cysteine acutely infracted rat heart model is also much higher than 99mTcglucaric acid .Thus 99mTc-cysteine could be used for the diagnosis of myocardial infraction in critical early hours.

Significant results were also obtained from projects implemented in the area of communicable disease. Studies shows that the biolarvicides Bacillus sphaericus and Bacillus thuringiensis treatment/toxin adversely affected the development of the filarial parasite (Wauchereria boncrofti) in the Culex.quinquefasciatus. They induced production/enhancement of humoral defense proteins or lectins in the mosquitoes, which affected the development of the parasite adversely. The mosquito pupicidal factor (proteins) has already been purified from the culture filtrate of Pseudomonas fluorescens mw~35kDa and 55 kDa. These protein were active against pupae of the three species of mosquitoes tested viz. Culex quinequefasciautus, Anopheles stephensi and Aedes Aegypti. Characterization of mosquitocidial proteins showed that they are thermostable glycoproteins. The pathogenesis of anemic condition with an emphasis on the factors leading to the reduced survival of red cells during visceral leishmaniasis infection has been elucidated. Results of a study on the role of Ureaplasma urealyticum in prematurity and diseases in newborn shows that the presence of U. urealyticum in lower respiratory tract at birth is associated with significant neonatal morbidity and the conformation of U. urealyticum in lower respiratory tract infection.

New diagnostic methods have been evaluated using ELISA, flow cytometery, RTPCR for the diagnosis of Chlamydia trachomatis and other STD pathogens. Cytokine levels in lower and upper genital tract in Chlamydia trachomatis infected women has been evaluated. Results of studies on development on novel anti-HIV molecules based on characterization of mechanism of action of Topoisomerase II inhibitor during HIV replication reveals that a novel class of ferrocene compound show significant Topo II poisoning and anti-HIV activity and can be exploited for their therapeutic potential in chronically HIV-1 infected patients.

An attempt has been made for the first time to characterize the PKDL parasite of Indian origin using APPCR fingerprinting methods. The study demonstrated that polymorphic AP-PCR fragments incurred from PKDL isolates are being transcribed. Four of the polymorphic AP-PCR fragments corresponded to genes that showed differential expression in pro and amastigote stages. Two of genes coding for Phosphodiestrase and DEAD box RNA helicases have been studied for the first time in Leishmaina donovani and both of these molecules are the parasite survival and can be a good drug target and / or vaccine candidate. One of the poliymorphic gene fragments coded for FeSOD, which was cloned and the recombinant protein product expressed in E.coli. It was shown to generate humoral immune response in the KA and PKDL patients as the antibodies against this molecule were detected in the patients' sera.

Plant Sciences

In the area of Plant Sciences several projects have been supported during the year. These include basic research and some application oriented R&D projects. Some of the projects funded during the year were:

Diseases of Certain Medicinal Herbs in Bhadra Wild Life Sanctuary, Karnataka; Histological, Biochemical & Control Studies on Timber Degrading Fungi of Gujarat; Search for bioactive molecules from genus Ganoderma (polyporaccae) occurring in south India; Cultured diversity of anoxygenic phototrophic rhizobacteria of paddy: Their geographical distribution, localization of species and plant growth promoting activity; PCR finger printing and rDNA sequencing to reveal genetic variations between the arbuscular mycorrhizal fungi from heavy metal contaminated soils; Physical mapping of simple sequence repeate (SSRs) in bread wheat; Quantitative Assessment of Status of Endemic, Rare, Endangered and Threatened Plants (ERETs): Developing a Protocol using Niche Modelling; Further studies on Indian Zingiberaceae; Generation of genetic variability through somclonal variation and induced in vitro mutagenesis for improvement of Rose-scented Geranium an exclusively vegetatively propagated aromatic plant; Oxidative stress responses to water deficit in different mulberry cultivars; A role for ATP- sulfurylase in the protection of photosynthesis and oxidative stress in mustard under salinity stress; Standardization of conditions for exploitation of spent substrate for decolorization of coloring dyes; Oxidative stress and Inducible tolerance mechanisms in rice plants under Aluminum toxicity; Molecular systematics of the genus Sapindus L. (Sapindaceae) in India using PCR- based techniques etc.

Some Important Results in ongoing projects were as follows:

In a project on "Molecular analysis of differentiation in the cotyledons of Vigna radiata", a unique differential regeneration response of the two cotyledon types, 'Cot E' (attached to the embryonal axis) and 'Cot' has been reported earlier in Vigna radiata (Fig. 5). During the tenure of the project the differential regeneration response in these two cotyledon types at the biochemical level and differential gene expression in 'Cot' and 'Cot E' during in vitro differentiation have been demonstrated. It has been predicted that perhaps the invertase enzyme (EC 3.2.1.26) expression during in vitro meristem/shoot differentiation mimics the event that takes place during seed germination.

In another ongoing project on "Molecular basis of unique develomental biology of Podostemads", the studies have been done on Dalzellia zeylanica and Polypleurum stylosum which belong to highly enigmatic family of aquatic angiosperms - the Podostemaceae. The plants grow attached to rocks in the river water with the help of adhesive hair present on the ventral surface. The vegetative plant body is rootless, highly crustose, dichotomously branched and bears marginal and dorsal leaves (Fig. 6). The solitary, terminal, trimerous flowers arise in cupules. The plants are predominantly self-pollinated and the pollen germinate immediately upon landing on the stigmatic papillae. The pollen tubes travel through the short stigma and inconspicuous style into the ovary. Although the pollen is shed at two-celled stage, the generative cell divides within the pollen tube to form two male gametes. After traversing through the placenta, the pollen tubes reach the mature ovules. A mature embryo sac is 4-nucleate, 4-celled. Prior to fertilization, the egg apparatus consists

Fig. 5. Northern blot of differentiating ‘Cot’ and ‘Cot E’, showing differential expression of invertase, corresponding stages of differentiation under the induction of cytokinin at the 6 and 9 days.

of two synergids and an egg cell. The three antipodals and a polar nucleus are absent. Out of the two sperms, one of them fuses with the egg cell whereas the second one does not fuse with the single polar nucleus, thus bringing about only syngamy or single fertilization, a feature unique to the family Podostemaceae.

Since the members of the family Podostemaceae represent novel developmental abnormalities, it was to analyze them at the molecular level and identify the genes that define their unique pattern of development. In order to characterize pattern formation genes, a cDNA library was made from young seedlings of Polypleurum stylosum, another member of the Podostemaceae. A total of 4253 high-quality sequences were obtained by ABI3700 automated sequencer. These ESTs could be assembled in to 429 contigs and 1654 singletons, representing unigenes. The BLASTX search led to identification of putative Arabidopsis orthologs for 1456 Polypleurum unigenes. The genes encode proteins involved in metabolism, response to stress, regulation of physiological processes, photosynthesis, and embryonic development. One of these is homologous to Arabidopsis MONOPTEROS (an auxin response factor, ARF5) protein sequence, a transcription factor mediating embryo axis formation. Further work is in progress to identify genes that express differentially and regulate Polypleurum development.

In another ongoing project on "Regulation Of Gene Expression In Light Signal Transduction Pathways In Arabidopsis thaliana", to understand the molecular basis of photomorphogenesis, a bHLH transcription factor (ZBF1/MYC2) has been characterized earlier that acts as a negative regulator of blue light mediated seedling development in Arabidopsis. Recently ZBF2 (a bZIP transcription factor; GBF1), another Z-box binding factor obtained in the ligand binding screen has been characterized. The DNA-protein interaction studies reveal that ZBF2/GBF1 interacts with the Z- and G-box light responsive elements of light regulated promoters. Genetic analyses of gbf1 mutants and over-expression studies demonstrate that GBF1/ZBF2 acts as a repressor of blue light mediated inhibition in hypocotyl elongation (Fig. 7), however, it acts as a positive regulator of cotyledon expansion during photomorphogenic growth. Furthermore, whereas GBF1/ZBF2 acts as a positive regulator of lateral root formation, it differentially regulates the expression of light inducible genes. Taken together, these results demonstrate that GBF1/ZBF2 is a unique transcriptional regulator of light signaling in Arabidopsis.

Fig. 6. A - Part of the thalloid plant of Dalzellia zeylanica showing flower buds.
B - Part of the thalloid plant of Dalzellia zeylanica showing flower buds with elongated pedicels.
C - Flowers of Dalzellia zeylanica depicting various stages of pollination.
D - Part of the thalloid plant of Dalzellia zeylanica with numerous pollinated flowers.

Fig. 7. Mutation in ZBF2/GBF1 results in growth defects in Arabidopsis seedling. Wild type (Col) and zbf2-1/gbf1-1 (Col) mutants are shown on the left and right, respectively, in each panel. A-F, Six-day-old seedlings were grown in constant darkness, White (5 ?mole m-2 s-1), White (30 ?mole m-2 s-1), Blue (30 ?mole m-2 s-1), Red (95 ?mole m-2 s-1), or Far-red (90 ?mole m-2 s-1), lights, respectively. The mutant seedlings display short hypocotyls and small cotyledons compared to corresponding wild type seedlings grown in white and blue lights specifically.

To decipher whether the components of light signaling defined by the constitutively photomorphogenic (pho) mutants identified influence the Z-box activity, Z-box containing synthetic (Z/NOS101-GUS, Z-GATA/ NOS101-GUS), GATA-box containing (GATA/NOS101-GUS) and native (CAB1-GUS) promoters were introduced into pho1 and pho3 mutant background by genetic crosses. The F1 plants were selfed in successive generations and the F3 and F4 seedlings grown in light and dark (for 6 days) analyzed for GUS activity. These results indicate that, for the Z-box and GATA-box promoters, GUS activity was high in both the pho mutant backgrounds when compared with the wild-type seedlings; in dark, the Z-box and GATA-box activity was relatively low. Z-GATA and native CAB1 promoters did not reveal much significant difference in GUS activity in the mutant background. These results indicate that the products of PHO genes are negative regulators and influence the Z-box and GATA-box activity.

In a on-going project on "Assessment of genetic variability in Tamarind (Tamarindus indica L) by using morphological traits and molecular markers", it was observed that all the characters varied significantly, except thickness of pod, seed to pod ratio, number of seeds per pod, seed weight per pod, stamen length, filament length, and style length. The variability noticed was narrow at molecular level compared to the variability at phenotypic level. Four elite tamarind genotypes are identified which are productive and regular bearing.

Fig. Assessment of Genetic Variability in Tamarind using molecular makers

Fig. Assessment of Gentic variability in Tamarind using morphological traits.

Variation pod bearing habit

Fig. Variations in Tamarind colour of pulp, seed size and pod length.

Biochemistry, Biophysics, Molecular Biology and Microbiology

As part of promotion of basic research in the area, 62 new research projects, with commitment of approx. Rs 14 crore for a period of 3 years, were granted to various organizations. Grants of approx. Rs. 7 crore were also disbursed for these projects. Out of these 62 projects, 35 research projects (with commitment of approx. Rs 8 crore for 3 years) were sanctioned to the academic sector which constitutes approx. 57% of the total research projects/grants provided during the year 2006-07.

The broad subject areas supported were:

• Structural biology: crystal structure determination of proteins; protein structure, function, folding and stability; protein-protein interactions to explore novel function(s) of bacterial haemoglobin, studies on plant lectins; folding of sweet protein monellin, structural proteomics of seed proteins with allergenic activity, characterization of silk protein, structural and functional studies for crystalline super fold, structural analysis of FAD synthease from yeast etc.
• Molecular and Cell biology: molecular basis of protein expression; regulation of protein synthesis;plant and stress biology; studies on non coding genes; mining of human genome for global regulatory genes, nature and mechanism of genomic imprinting, studies on pancreatic cancer genes, chromosome recombination studies using yeast etc.
• Microbiology: microbial biodegradation; isolation of enzymes from microbes; molecular cell biology of pathogens: Plasmodium, Leishmania, Pseudomonas, mycobacterium etc. apoptotic and anti-apopotic signalling in Mycobacterium tuberculosis infected macrophages and dendritic cells, Regulation of Carbon & nitrogen metabolism in Saccharomyces cerevisiae etc.

A number of collaborative research projects were also funded, which are:

• "Molecular mechanism of cold acclimation in a Cyanobacterium synchocystis: role of molecular chaperons during cold acclimation" at Centre for Cellular and Molecular Biology, Hyderabad and University of Hyderabad, Hyderabad;
• "Mining the human genome for global regulatory genes: identification, characterization and functional analysis" at Dr B R Ambedekar Centre for Biomedical Research, Delhi and Centre for Cellular and Molecular Biology, Hyderabad.
• "Molecular characterization of genes causing primary angle glaucoma in Indian population" at L V Prasad Eye Institute and CCMB, Hyderabad.
• "To study the effect of small interfering RNA (siRNA) on the EpCAM expression of Y79 retinoblastoma cell line and its correlation with invasion and migration properties of cell line" at Vision Research Foundation, Sankara Nethralaya, Chennai and Indian Institute of Technology, Madras, Chennai.
• "Design, synthesis and characterization of á,â-dehydrophenylalanine containing peptide based inhibitors of human islet amyloid polypeptide fibrillogenesis" at International Centre for Genetic Engineering and Biotechnology(ICGEB) New Delhi and Indian Institute of Science, Bangalore.

Accomplishments: The programme has been successful in consolidation of Indian researchers' competence in the area of structural biology, molecular and cell biology and microbiology by enabling training in emerging areas and publication of research papers which will provide new knowledge and vital research leads in the field.

Research highlights and achievements from some of the successfully completed /ongoing projects are:

Inhibitors of the enzymes essential for infectivity and survival of protozoan parasite leishmania were identified.

• Calcium mediated intersubunit interactions having a vital role in stability of Sesbania mosaic virus capsid were demonstrated.
• A Transposon Display (TD) method for identification of strain variation in Entamoeba histolytica was demonstrated..
• Role of cAMP in growth, morphogenesis and cell cycle regulation in Candida albicans was proposed.
• In a study on mechanism of action of the DNA repair gene rhp6, a mediator of Rhp6- called Uhp1 (ubiquitinated histone like protein) was identified that seemed to play a transient role in reestablishment of the chromatin structure during switching.
• Mechanism of apoptotic induction by S29 ribosomal protein and possibility of S 29 ribosomal protein as a target for novel anticancer therapy was suggested.
• Expression level of p14, p15, p16 and p53 tumor suppressor genes and oncogenes was determined in some eastern India population.
• Several genes (CvSox9 and CvDmrt1), were identified with strong indications that these genes may be involved in testis differentiation in the lizard, Calotes versicolor. A candidate gene, CvWnt4, for female pathway was identified.
• A substantial amount of free soluble p-hydroxybenzoic acid was detected from the hairy root cultures of D. carota, providing important new clues for generating phenolic antioxidant and flavor precursors.
• Structural studies on plant lectins: crystallographic and modeling studies on unusual sugar specificity of banana lectin, specificity of basic winged bean lectin for Tn antigen, galactose specific lectin from Dolichos lablab were undertaken.
• A number of genes and their promoters involved in signalling under salinity stress in Oryza sativa were identiifed .
• Winged bean chemotrypsin/trysin inhibitor (WBCTI) gene was cloned for the first time and sequence submitted to the GeneBank.

Future plan: With a view to improving the overall performance and consolidation of Indian researchers' competence in the area - by rectifying weaknesses; building on our strengths; and initiating new lines of endeavors - the following areas of research during the XI plan were identified:

• The areas requiring special attention were identified as: classical genetics and evolutionary biology; taxonomy; physiology; immunology; microbiology; plant biology; cancer biology, neurobiology and reproductive biology.
• The areas of our strength which need further strengthening were identified which include: structural biology; biophysical chemistry; and conventional biochemistry.
• Emerging areas which need to be vigorously pursued were identified as: Genomics and Proteomics; structure based ligand design for biological macromolecules including drug targets; systems biology; nanoscience; nanotechnology and single molecule biology; stem cell biology
• Industry-academia interaction in applications such as biodegradation and biofuels also need to be addressed.

To facilitate and strengthen research, setting up of new facilities during the 11th plan were recommended which include: High power photon source, Central microbial stock and identification facility including those for pathogens, Central resource facility for animals including transgenic ones, Centre for nanobioscience and nanobiotechnology, Centre for molecular medicine, National library of bioactive compounds, Joint venture with industry for biologicals.