IERG Abstracrt Book.indd - LV Prasad Eye Institute

64 Basic Poster SessionsMethods: A prospective nonrandomized investigation was undertaken to evaluate 33 eyes of33 patients with clinically suspected post cataract endophthalmitits. Vitreous tap was collectedfrom these patients. Besides routine bacterial culture and sensitivity, vitreous samples wereevaluated by RT-PCR. Real Time PCR using Taqman based chemistries targeting the 16S rRNAgene using universal primer and universal probe was performed. The positive samples werefurther sequenced by using Big dye terminator V.3.1 followed by BLAST analysis for speciesidentification.Results: Of 33 presumed cases of postoperative endophthalmitis (33 untreated), RT-PCRshowed positivity in 60% (20 eyes) and culture in 45% (15 eyes). The sensitivity and specificityof RT-PCR was found to be 60% and 100%. Thirteen eyes showed negativity in 39% by cultureas well as by RT-PCR. The time taken for RT-PCR assay was 2 to 3 hours, whereas positivebacterial culture took atleast 1 to 2 days.Conclusions: The Real Time PCR not only proved an effective rapid method for the diagnosisof postoperative endophthalmitis, but also more sensitive than conventional methods. Barringthe potential limitations, RT-PCR can be a fast diagnostic tool and may be useful as an adjunctto identify potential pathogens.IBP 007Effect of Endoplasmic Reticulum Stress on the Lens Epithelial CellsDevki Sheth, AR Vasavada, SR Kaid JoharIladevi Cataract and IOL Research Centre, Ahmedabad, IndiaPurpose: To study the effect of Endoplasmic Reticulum (ER) stress on proliferation,differentiation and cell death of cultured Lens Epithelial Cells (LECs).Materials: Primary culture of LECs was carried out and ER stress was induced in 70%confluent LECs using 1 um tunicamycin for 24 hrs. Proliferation of cell was detected usingimmunofluorescence localization of BrdU. LECs differentiation was detected using antibodiesagainst pax6, alpha smooth muscle actin (aSMA), E-cadherin and beta-catenin. TUNEL assay wascarried out to detect the apoptotic cells and single cell gel electrophoresis (SCGE) assay tomeasure extent of DNA damage in individual LECs.Results: ER stress induced LECs were 3 times less responsive to proliferation compared tothe control group. The TUNEL positive cells and the extent of DNA damage as indicated bythe image analysis of tail length obtained from SCGE assay were more in the ER stress inducedcells than that of control cells. The differentiation studies indicate that the LECs markers weremore expressed in ER stress induced cells than control cells.Conclusions: ER stress maintains LECs in undifferentiated cells along with decreasedproliferation and increased apoptosis.