Balkan Journal of Medical Genetics

INCREASED EXPRESSION OF CARDIOTROPHIN-1 IN CARDIOMYOPATHY PATIENTS
Sharif S1,*, Saleem A1, Naz S1, Rashid F1, Iqtedar M2, Kaleem A2, Latif A1
*Corresponding Author: Dr. Saima Sharif, Department of Zoology, Lahore College for Women University, Jail Road, Lahore, Pakistan. Tel: +92-333-409-2232. Fax: +92-042-9920-3077. E-mail: saimasharif04@ gmail.com
page: 21

METHODOLOGY

The cross-sectional study was conducted at the Fatima Memorial Hospital (FMH) of Lahore, Pakistan. Permission for sampling was obtained from the Ethics Committee of the institution. Samples were collected from the Cardiology Department of FMH between January and June 2016. The sample size was calculated using sample calculator on Raosoft Inc. (http://www.raosoft.com/ samplesize.html) with 5.0% margin of error and 50.0% confidence interval with expected prevalence of cardiac diseases as 410/ 100,000 in our population. The study population was divided into two groups: the control group consisted of 20 healthy individuals and the CM group included 20 subjects. Inclusion and exclusion criteria were made for appropriate selection of patients which were as follows: patients with suspected HF, patients with left ventricular (LV) dilatation and dysfunction were included in the study. Other acquired or congenital cardiac diseases such as myocardial infarction, other coronary vascular disease, myocarditis, pericardial diseases (not mild pericardial effusion that may be secondary to HF) and patients of any cardiac/genetic disease were excluded from the study. Written informed consent was obtained from the subjects before their participation in the study. A structured questionnaire used for data collection regarding age, gender, habits, duration of disease and family history of disease. A blood sample (3 mL) was drawn from the enrolled subjects and transferred immediately to EDTAcontaining vacutainers and mixed gently for 1 min. to prevent blood clotting and inhibiting activity of nucleases. The sequence of human gene CT-1 was retrieved from the National Center for Biotechnology Information (NCBI) (www.ncbi.nlm.nih.gov) and the BLAST (basic local alignment tool) in the bioinformatics tool (www.genome. ucse.edu). Only exomic sequences (NM-001330.5) were used for designing the primers (www. genome.ucse.edu). RNA was isolated by the trizole method [10]. RNA quantity and quality was determined using the Nano Drop™ 2000/2000c spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Then, cDNA synthesis was done by the reverse transcriptase method. Furthermore, cellular gene expression was determined by reverse-transcriptasepolymerase chain reaction (RT-PCR). Statistical analysis was done using the Statistical Package for the Social Sciences (SPSS), version 23 (IBM Corporation, Armonk, NY, USA). The statistical difference between the groups was analyzed by the Student’s t-test. Pearson correlation was done to determine the relationship between the CT-1 gene and demographic characteristics.

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