IERG Abstracrt Book.indd - LV Prasad Eye Institute

Basic Poster Sessions67when treated with SERCA inhibitor, Thapsigargin, calcium patterns are affected when thedistribution of the intracellular RyR, is perturbed. We showed that the specific activities ofRyR markedly increased in the low-density region of the gradient compared with controls. Incontrast Cyclopiazonic acid (CPA), another SERCA inhibitor does not affect RyR or calciumdynamics when co-treated with methyl-cyclodextin (ßMCD), which depletes cholesterol anddisrupts membrane. Disruption by ß-MCD altered the distribution of caveolin-1, a markerfor lipid raft, and RyR to non-raft fractions with higher densities. ßMCD- induced disruptionof rafts inhibited agonist-evoked Ca2+ wave propagation in primary bovine Muller glia andattenuated wave speed.Conclusions: These results indicate that in glial cells, Ca2+ signaling proteins exist in organizedmembrane microdomains, and these complexes include proteins from different cellularmembrane systems. Such an organization is essential for Ca2+ wave propagation.IBP 012Biophysical Characterization of Human Myocilin and the C-Term RegionK Rangachari, P J Eswari Pandaranayaka, J Jeyalaxmi, S Krishnaswamy, Prasanthi Namburi, PSundaresan, S R KrishnadasDept. of Genetic Engineering, School of Biotechnology, Madurai Kamaraj University, Madurai,Tamilnadu, India.Purpose: To study, using biophysical methods the human myocilin and the C-term region.Methods: Myocilin (55 kDa mol wt, 504 aa, Swissprot Q99972) is localized both intracellularlyand extracellularly at multiple sites and may exert diverse biological functions. Based on amodel of myocilin built by us, four regions have been identified. These regions are N-term,coiled coil, hinge and C-term. The C-term region contains the olfactomedin domain. This regionhas been cloned in E.coli for over expression.Result: In order to study the structural features of the olfactomedin domain containing regionwe have over expressed it in E.coli by using pET-20b+ vector, purified, optimized refoldingconditions. Biophysical studies have been done on full length human myocilin which has alsobeen purified.Conclusions: The C-term region of Myocilin has been purified and characterized biophysicallyand needs to be further studied. The characterization of the full length myocilin usingspectroscopic and light scattering studies suggests the presence of beta sheet region and thetendency to aggregate.IBP 013Identification of Fusarium Species by Molecular Methods and their AntifungalSusceptibility from Patient’s with Corneal UlcerLakshmi Priya Jeganathan, N V Prajna, Lalitha PrajnaDr G Venkataswamy Eye Research Institute, Aravind Medical Research Foundation,Anna Nagar, Madurai, India.Purpose: To investigate the fungal pathogens by molecular techniques and to analyze theantifungal susceptibility from clinical isolates. Identification of a pathogen at the species levelwill be helpful in better diagnosis which aid in therapy and also for the clinical outcome andsource of infection.