International Workshop on Molecular Biology Techniques Related to Infectious/Genetic Diseases & Human Identification
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Item Molecular diagnosis of Down syndrome cases(Faculty of Medicine, University of Kelaniya, 2015) Podder, G.; Madhusnata, D.Introduction: Down Syndrome (DS), the most frequent form of mental retardation, is characterized by well-defined and distinctive phenotypic features. Research DS-related genes is based on studying the genes located on chromosome 21. Understanding the genes involved may help to target medical treatment to individuals with DS. It is estimated that chromosome 21 contains 200 to 250 genes. It has been identified major genes involved in DS characteristics are normally in the region 21q21–21q22.3. Methodology: Blood samples from 85 DS cases and 30 normal cases were analyzed for D21S11 marker gene. Fluorescent dye-labeled primers were used in PCR amplification of this marker. The PCR amplified product was analyzed by GeneScan software version 4.1 automatically. Results: The short tandem repeat marker frequency was estimated in triallelic, diallelic, and monoallelic patterns. Seventy nine (92.94%) of 85 DS cases showed 3 distinct peaks for D21S11 marker and 5 (5.88%) DS cases showed two uneven peaks. One DS (1.17%) sample showed only one large peak. For healthy individuals, two peaks were observed in all cases. Discussion: Polymorphic STR DNA markeris useful for determining the numbers of chromosome 21. The high sensitivity and automation of the procedures suggest a good prospect for use of this method in detection of DS. However, this is a preliminary investigation and a large-scale study is necessary to validate the clinical application of this protocol.Item Study on predisposing factors of type 2 diabetes mellitus(Faculty of Medicine, University of Kelaniya, 2015) Mukherjee, P.B.; Madhusnata, D.Introduction: Type 2Diabetes Mellitus (T2DM) is a chronic progressive disease. There is increasing evidence that T2DM has been associated with elevated levels of DNA damage and a decreased efficacy of DNA repair causing genomic instability and consequently cancer. The frequency of Micro Nuclei (MN) has been mainly used as a biomarker to evaluate genotoxic risks in the environment. Oxidative Stress (OS) plays a critical role in the pathogenesis of both types of DM and its complications. OS causes oxidative degradation of lipids in cell membranes (lipid peroxidation) resulting in cell damage. In order to combat the cell damage, body has antioxidant property as a defence mechanism. Over production of ROS (OS) alters the OS biomarkers. Vascular Endothelial Growth Factor (VEGF) plays a key role in the pathogenesis of different complications of T2DM. Polymorphisms in VEGF gene contribute to the risk of diabetic complications. Objectives: To determine lipid peroxidation and antioxidant (catalase) activities, frequency of MN and molecular analysis for diagnosis for the progression of diabetic complications in association with genetic polymorphisms in VEGF gene. Methodology: Lipid peroxidation and catalase activity are assayed spectrophotometrically in both diabetic cases and non diabetic controls. The slides for MN analysis in buccal cells are stained with giemsa and scored randomly. DNA from peripheral blood are isolated and amplified by PCR followed by RFLP. Results: Lipid peroxidation is higher than the catalase activity in diabetic samples rather than the non-diabetic controls. The frequency of MN is also elevated in diabetic samples. Polymorphisms in VEGF gene have been found in diabetic cases. Discussion: Elevated lipid peroxidation than the catalase activity signifies increase in cellular susceptibility to oxidative stress. Increase in MN frequency in diabetic patients denotes a risk for causing cancer. Polymorphisms in VEGF gene state its association for the progression of late diabetic complications.