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Innovation Project : SVC 101

Survey of Trees in Delhi Region and Screening of Selected Medicinal Plants for Anti-cancer Activity
Project Hypothesis

India enjoys a rich natural biodiversity and offers unique opportunities for the discovery ofnew drugs. Medicinal plants play a dominant role in this discovery, as over 60% of approveddrugs or those in late stages of development are of natural origin. However, in recent years, ahuge loss of plants has been reported - a loss in forest cover by an area of 0.38 km22006 and 2011 - in Delhi, mainly due to development in urban areas. As this was a staggeringfigure, this prompted the investigation of what species of trees are actually present and whichones may have become extinct during the last 50 years. Therefore, there is an utmost need tosurvey plants, particularly trees, of Delhi.
Meanwhile, cancer is emerging as a major public health problem in India. While mostcancers are tobacco-related and are preventable, the cancers in women, such as cervical andbreast cancer are not easily preventable. There is considerable evidence of the potential ofmedicinal plants being used in various traditional, complementary and alternative systems forcancer treatment. The preliminary step in plant based anticancer drug development is screeningof the medicinal plants for anticancer activity.
Being an innovative project, it was appealing idea to conduct an interdisciplinary study onthe available tree species and to conduct tests on the plant parts to detect anticancer activity. Forthis, we have selected certain plant species that according to traditional knowledge or literature,such as Ayurveda, possess anti-cancer activity and on these plants we will carry out tests toverify the traditional claims.
The current study would be helpful to develop a list of tree species present in Delhi after theloss of biodiversity. The screening of the medicinal plants/ plant parts would be helpful for drugformulation against cancer. The crude extracts enriched with anti-cancer activity would befurther purified for the isolation of bioactive anti-cancerous compounds. After, isolation andcharacterization of active compounds would be studied with all the molecular targets responsiblefor cancer. Further, chemical analogs would be synthesized for all the identified anti-cancerouscompounds and used for commercial preparation of drugs for combating this deadly disease.
The three main objectives of the project are: to survey tree species in Delhi; to prepare thecrude extract from various parts (such as root, bark, stem, leaves, flowers, etc.) of selectedmedicinal plants using various solvents; and to evaluate these crude extracts for anti-canceractivity against certain cancer cells.

Innovation Project : SVC 102

Screening of Cyanobacteria for Compounds with Antimicrobial activity and explore its Biosynthesis Regulation by Heterochromatin

Cyanobacteria constitute a unique group of oxygenic photosynthetic prokaryotes. They occupy diverse habitats and produce an array of secondary metabolites including alkaloids, isoprenoids, polyketides and non-ribosomal peptides having broad spectrum of activities. To mention a few antitumor, antibacterial and antiviral effects and protease inhibition. Their potential as a good source of new therapeutic lead compounds has been realized during the last two decades. Various studies have focused on the typically encountered environmental factors (temperature, light, minerals such as nitrogen and phosphate) that may influence changes in the production of secondary metabolites. Recent research has identified the presence of several ncRNAs in cyanobacteria which are known to regulate stress response. In response to regulated changes in gene expression, the secondary metabolisms are redirected to cope with the undesired effects of the hostile situation. This is achieved by up-regulating the synthesis of proteins and metabolites. The Present work was under taken with two prime objectives viz. (i) Screening of the cyanobacteria for antimicrobial activity, and (ii) To study the relation between bioactive molecule production under stressed environment and stress regulating non-coding RNAs.

Cyanobacterial samples collected from different sites of Delhi (Hauz khas, Indiagate, Satyaniketan, Nazafgarh, Ranikhera) and NCR (Manesar, Noida) along with purified strains of Anabaena and Microcystis were screened for antimicrobial activity. Since production of secondary metabolites is a factor of age, the exact cell age was estimated by growth behavior of cyanobacterial strains on the basis protein estimation. The specific growth rate was calculated according to Kartz and Mayer. Extracts from oven dry cyanobacterial biomass at 30 days, produced larger inhibition zones, reflecting that the duration was optimal for isolating the intracellular metabolites. During preliminary findings for solvent potential it was observed that methanolic extract had more bactericidal property than dimethyl sulfoxide and water. Bioassay for antibacterial property of methanolic extracts, using both pathogenic (Escherichia coli) and non-pathogenic (Enterobactar aerogenes) bacteria were done. It was observed that Anabaena collected from a water pond (Manesar) showed two times more bactericidal activity than the extract of purified strain aginst E. aerogenes. Microcystis from Manesar and Ranikhera johad (water pond used for multipurpose) expressed maximum activity while same genus from RaniKhera Devali (a temple compound water pond) and Huaz Khas pond did not show significant effect on E. aerogenes, suggesting that may be the production of bioactive molecules is niche and species specific. Against pathogenic bacteria (E. coli) maximum bactericidal potential was observed with purified Anabaena extract while lowest for Oscillatoria from Satya niketan. Studies on stress response of cyanobacterial biomass under different concentrations of nitrate, phosphate and sodium chloride were carried out by adding the mentioned component to the culture media (BG11 and CHU-10) and cells were cultured at 28oC under continuous irradiance with periodic shaking for 30 days. It was observed that there was significant effect on biomass production. The cyanobacterial biomass (stressed) samples were collected and further studies on expression level of along with interaction of regulating non-coding RNA under different stress condition are yet to be carried out at a later date.

Innovation Project : SVC 103

Project Title: Comparative Anti-oxidant Profiling of Various Indian Rice Cultivars in response to Salinity Stress

Salinity in soil or water is of increasing importance to agriculture because it causes a stress condition to crop plants. Particularly for rice (Oryza sativa L.), a species native to swamps and freshwater marshes,secondary Stalinization is becoming an increasingly serious production constraint (Akbar and Ponnamperuma, 1980). Because of the inherent sensitivity of rice plant to salt stress (Francois and Mass, 1994), there has been a great interest in developing varieties that are resistant to salinity. Defining salt tolerance,however, is quite difficult because of the complex nature of salt stress and the wide range of plant responses. Present proposal was designed to determine the effect of salt-stress on anti-oxidant enzyme activities,electrolyte leakage and Na+/K+ content of various Indian rice cultivars. In the present study, comparative effects of salt-stress have been investigated for rice [Indian cultivars] by considering anti-oxidant responses of young seedlings together with biochemical and physiological analyses like proline,chlorophyll estimation, electrical conductivity and Na+/K+ content. Results from this study have supplied information on the possible involvement of different anti-oxidative enzymes under salt-stress. Understanding the biochemical and physiological basis of salinity could help in selection and improvement of rice, which is an agronomically important plant.

Oryza sativa L. is one of the most important crops all over the world and is considered to be the primary staple food for half of world's population. Extensive research is being carried out worldwide for further improvement of rice cultivars to adapt according to the environment with high yield. Rice plant is known to be a glycophyte hence susceptible to salt stress. Exploring the physiological and biochemical mechanism of salinity could be helpful in the selection of rice cultivar for the agriculturists as well as breeders. There are few reports where salt tolerance mechanism in rice cultivars has been studied but these varieties have not been studied so far. Our preliminary findings indicate that there were substantial differences between the morphological, biochemical and physiological parameters of all Indian varieties studied. Salt sensitive varieties exhibited high Na+ accumulation, and increased activity of antioxidant enzymes like superoxide dismutase, glutathione reductase, catalase, peroxidise and proline as compared to salt tolerant varieties. In order to see the effect of salt stress in rice, various rice cultivars, Pusa sugandha, HBL24, IR 64 and Pokkali etc have been selected for this study. The present study indicated that salinity stress caused significant changes in morphological parameters including decreased root length, shoot length and fresh weight and activity of antioxidant enzymes in different varieties of Oryza sativa.L. Significant enhancement was detected in activities of all 4 enzymes as compared to control or Untreated plants. But there were substantial differences in Enzyme activity when compared between different varieties. Rice cultivars viz., PB-1, Pusa Sugandha, HBL-24, Parmal, CSR 30, PNR-162, Pusa-44 were screened for salt stress to measure electrolyte leakage and Na+ / K content. . Among these varieties we found that CSR 30 and HBL were semi-tolerant while rest were sensitive to salt stress because in these two varieties the Na+/K+ concentration were higher as compared to other and very similar to that of controlled samples. In case of electrolyte leakage, it was found that the change in the electrolyte leakage in these two varieties were very less as compared to the sensitive varieties. In case of sensitive varieties the sodium uptake was high as compared to the semi-tolerant varieties.This suggest that Plants of VarietyPokkali also known to be a Salt tolerant variety exhibited highest adaptive potential under salinity stress as evident by the changes in growth parameters and antioxidant defense mechanism as compared to other varieties. The above findings will be beneficial for agriculturists and biotechnologists to select the appropriate variety, either to be grown or genetically engineered.This project has cogitated deeply on raising awareness about a major problem the staple food of India is facing which is salinity. If the farmers are made aware of the correct classification between a salt sensitive rice variety and a salt tolerant rice variety, they can improve their farming techniques yielding better productivity. The plant biotechnologists can savoir the taste of research by following a reliable data and conceptualizing a way to convert a salt sensitive rice variety to a salt tolerant rice variety which will be of much profit to the agriculture and economic world. The Indian society would benefit from it because with the growing population the net rice productivity of the country will increase hence stepping closer to solving food crisis.

Innovation Project : SVC 104

Screening of Indian population for possible polymorphisms in candidate genes of extracellular matrix proteins that could lead to Disc degeneration leading to herniation

Department of Biochemistry
Project Investigators : Kavitha Rajsekar,Dr.Meenakshi Kuhar,Dr.Nandita Narayanasamy,Dr.Shalini Sen
Mentors: Dr. S. Rajasekaran (MS Ortho, PhD), Dr.Sandeep Vaishya (MS, Mch)
Students : Arshiya Bhatt, Parth Kapoor,Pallavi Deolal,Urmila,Mamta,Kashish Singh,Jasmeet Kaur,Pooja Badhwar,Palak,Anupam and Sonali

Project Summary

Basic science research has demonstrated that the intervertebral disc is largely an avascular tissue occupied by inadequately characterized cells in an extensive extracellular matrix network. The central gelatinous nucleus pulposus is contained within the more collagenous anulus fibrosus laterally and the cartilage end plates inferiorly and superiorly. The annulus consists of concentric rings or lamellae, with fibers in the outer lamellae continuing into the longitudinal ligaments and vertebral bodies. This arrangement allows the discs to facilitate movement and flexibility within what would be an otherwise rigid spine. Biochemically, the important constituents of the disc are collagen fibers, elastin fibers, aggrecan, glycoaminoglycans and other associate adhesion proteins like integrins and connexins.

Lumbar disc degeneration occurs commonly in humans. There are a variety of factors that contribute to this condition. The disc itself is an active tissue that contains significant mechanisms for self-repair. The strength of the lumbar disc is related to the fluid and proteoglycan content of the disc.

The proposed study aims to understanding the importance of ECM and cell adhesion molecules in disc cell integrity. The genetic variants will help us identify populations groups which can be susceptible to disc degeneration (if any) .The study also aims in understanding stress related factors associated with disc degeneration which could be an underlying cause for oxidative stress by checking levels of TNF-alpha, IL-6 and IL-1 in the plasma.

The biochemical events that occur with the 'aging spine' and in particular, the role of inflammatory mediators in intervertebral disc degeneration have not been studied assertively.No study till date has been reported in the Indian population. There are no molecular, biochemical, or biologic markers available for various levels of disc degeneration and serious study of disc-cell biology at this level would be most rewarding to start new ventures with progenitor stem cell research for disc cell replacements and replacements of extracellular matrix components. Stress is another major factor which could be a part of disc degeneration leading to herniation. Screening the normal and the diseased population for the presence of polymorphic variants of collagen and to assess the association of apoptotic induced disc herniation in relation to proinflammatory stress condition could be a new beginning to the therapeutic interventions for spinal disc problems.