Balkan Journal of Medical Genetics

ASSOCIATION OF GLUTATHIONE-S-TRANSFERASE (GSTM1 and GSTT1) AND FTO GENE POLYMORPHISMS WITH TYPE 2 DIABETES MELLITUS CASES IN NORTHERN INDIA
Raza ST, Abbas S, Ahmad A, Ahmed F, Zaidi ZH, Mahdi F
*Corresponding Author: Syed Tasleem Raza, Ph.D., Molecular Biology Laboratory, Department of Biochemistry, Era’s Lucknow Medical College and Hospital, Hardoi Road, Lucknow, Uttar Pradesh, India 226025. Tel.: +91-522- 240-8122; 240-8123. Fax: +91-522-240-7824. E-mail: tasleem24@gmail.com
page: 47

MATERIALS AND METHODS

Patient Selection. A total of 101 blood samples from T2DM patients and 97 healthy controls were collected at the Diabetic Clinic, Department of Medicine of Era’s Lucknow Medical College & Hospital, Lucknow, Uttar Pradesh, India. Written informed consent was obtained from all participants. Data collection was done for each patient on clinical variables including age, alcohol consumption, body mass index (BMI), height, weight, cigarette smoking, family history, etc., patients with overnight fasting plasma glucose of more than 126.0 mg/dL on two consecutive events were included in the T2DM category, while samples having a fasting blood glucose level below 110.0 mg/ dL without a family history of diabetes were included in the study as controls. Patients with type 1 presentation, defined as diabetic ketoacidosis, acute presentation with heavy ketonuria (>3+), or uninterrupted requirement of insulin within 1 year of diagnosis, were excluded. Ethical Committee clearances were obtained from the respective departments, prior to the recruitment of subjects in this study. Biochemical Estimations. The BMI was calculated according to Quetelet equation by using weight in kg, height in m2. Fasting plasma glucose and random blood sugar (glucose oxidase-peroxidase method). Serum creatinine concentration was assessed by a kinetic Jaffe’s method. Serum cholesterol (cholesterol oxidase-peroxidase), serum triglyceride (glycerol phosphate oxidase-peroxidaseamidopyrine method), high-density lipoprotein (HDL) cholesterol (immunoinhibition) were assessed by XL-300 Transasia Fully Auto Analyzer; Transasia, Mannheim, Germany. Low-density lipoprotein (LDL) cholesterol levels were calculated using the Friedewald formula [19]. Very low-density lipoprotein (VLDL) cholesterol was determined by an enzymatic method. Hb A1C was measured using a semi auto analyzer (Transasia). DNA Extraction. Peripheral blood (5 mL) was collected from all the subjects in 0.5 M vacutainers with EDTA as anticoagulant. Genomic DNA was isolated from whole blood using the standard phenol- chloroform extraction method [20]. The DNA concentration was determined by spectrophotometer and stored at –20 °C.

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