Balkan Journal of Medical Genetics

POLYMORPHISM OF ANGIOTENSIN-CONVERTING ENZYME (rs4340) AND DIABETIC NEPHROPATHY IN CAUCASIANS WITH TYPE 2 DIABETES MELLITUS
Šeruga M, Makuc J,, Završnik M, Cilenšek I, Ekart R, Petrovič D M. Šeruga and J. Makuc contributed equally to this study.
*Corresponding Author: Professor Dr. Daniel Petrovič, M.D., Ph.D., Institute of Histology and Embryology, Faculty of Medicine, University of Ljubljana, Korytkova 2, 1000 Ljubljana, Slovenia. Tel: +386-1-543-7360. Fax: +386-1-543-7361. E-mail: daniel.petrovic@mf.uni-lj.si
page: 29

MATERIALS AND METHODS

We enrolled 651 unrelated Caucasians with T2DM from outpatient clinics of the University Medical Centre Maribor and the General Hospitals in Murska Sobota and Slovenj Gradec, Slovenia. Two hundred and seventy-six patients also had DN (cases) and 375 subjects with T2DM of more than 10 years duration but no clinical signs of DN (controls) were enrolled in the study. Diagnosis of DN was made according to the World Health Organization 1999 diagnostic criteria [16]. We excluded patients with overt nephropathy, poor glycemic control, significant heart failure New York Heart Association classification II-IV (NYHA II-IV), alcoholism, infection and other causes of renal disease. The study was approved by the National Medical Ethics Committee and was performed in compliance with the Helsinki declaration. After informed consent for participation in the study was obtained from all patients, a detailed interview was made. Biochemical Analyses. Total serum cholesterol, low-density lipoproteins (LDL), high-density lipoproteins (HDL), triglycerides (TGs), serum cystatin C, fastingserum glucose, serum glycemic Hb (Hb A1c), serum urea, serum creatinine were determined by standard biochemical methods. Albumin-to-creatinine ratio was also determined for each patient in three urine samples, according to the diagnostic criteria. To assess the kidney function, we used the modification diet in renal disease (MDRD) equation and serum-cystatin C [21,22]. Genotyping. Genomic DNA was extracted from 100 μL of whole blood using a Qiagen isolation kit (Qiagen GmbH, Hilden, Germany) following the blood and body fluid spin »V3« protocol. The protocol was supported by five different reagents (QIAgene DNA Blood Mini Kit; Qiagen GmbH): buffer AL, 96.0% ethanol, buffer AW1, buffer AW2, buffer AE and appropriate amount of proteases (285 μL of proteases/200 μL of blood). From 200 μL of blood, 3-12 mg of genomic DNA, i.e., 30-40 ng/ mL were extracted according to the instructions of the manufacturer. The protocol for rs4340 polymorphism of the ACE gene was as follows: one cycle at 94 °C for 5 min. followed by 30 cycles (94 °C for 60 seconds, 58 °C for 60 seconds and 72 °C for 60 seconds) and finishing with one cycle at 72 °C for 5 min. Statistical Analyses. Statistical analyses were conducted with the use of the Statistical Package for the Social Sciences program for Windows, version 20 (SPSS Inc., Chicago, IL, USA). Continuous clinical data were compared by unpaired Student’s t-test, while the χ2 test was used to compare discrete variables. Data were expressed as mean ± SD (standard deviation) (continuous variables) or as the number and percent of patients (categorical variables). All variables that showed significant differences by univariate analysis (with a p values of <0.05 were considered significant) were analyzed together using a logistic regression method. A p value of <0.05 was considered statistically significant. Deviation from Hardy-Weinberg equilibrium (HWE) was assessed by the exact test (http://ihg. gsf.de/).

Number 27 VOL. 27 (2), 2024	Number 27 VOL. 27 (1), 2024
Number 26 Number 26 VOL. 26(2), 2023 All in one	Number 26 VOL. 26(2), 2023
Number 26 VOL. 26, 2023 Supplement	Number 26 VOL. 26(1), 2023
Number 25 VOL. 25(2), 2022	Number 25 VOL. 25 (1), 2022
Number 24 VOL. 24(2), 2021	Number 24 VOL. 24(1), 2021
Number 23 VOL. 23(2), 2020	Number 22 VOL. 22(2), 2019
Number 22 VOL. 22(1), 2019	Number 22 VOL. 22, 2019 Supplement
Number 21 VOL. 21(2), 2018	Number 21 VOL. 21 (1), 2018
Number 21 VOL. 21, 2018 Supplement	Number 20 VOL. 20 (2), 2017
Number 20 VOL. 20 (1), 2017	Number 19 VOL. 19 (2), 2016
Number 19 VOL. 19 (1), 2016	Number 18 VOL. 18 (2), 2015
Number 18 VOL. 18 (1), 2015	Number 17 VOL. 17 (2), 2014
Number 17 VOL. 17 (1), 2014	Number 16 VOL. 16 (2), 2013
Number 16 VOL. 16 (1), 2013	Number 15 VOL. 15 (2), 2012
Number 15 VOL. 15, 2012 Supplement	Number 15 Vol. 15 (1), 2012
Number 14 14 - Vol. 14 (2), 2011	Number 14 The 9th Balkan Congress of Medical Genetics
Number 14 14 - Vol. 14 (1), 2011	Number 13 Vol. 13 (2), 2010
Number 13 Vol.13 (1), 2010	Number 12 Vol.12 (2), 2009
Number 12 Vol.12 (1), 2009	Number 11 Vol.11 (2),2008
Number 11 Vol.11 (1),2008	Number 10 Vol.10 (2), 2007
Number 10 10 (1),2007	Number 9 1&2, 2006
Number 9 3&4, 2006	Number 8 1&2, 2005
Number 8 3&4, 2004	Number 7 1&2, 2004
Number 6 3&4, 2003	Number 6 1&2, 2003
Number 5 3&4, 2002	Number 5 1&2, 2002
Number 4 Vol.3 (4), 2000	Number 4 Vol.2 (4), 1999
Number 4 Vol.1 (4), 1998	Number 4 3&4, 2001
Number 4 1&2, 2001	Number 3 Vol.3 (3), 2000
Number 3 Vol.2 (3), 1999	Number 3 Vol.1 (3), 1998
Number 2 Vol.3(2), 2000	Number 2 Vol.1 (2), 1998
Number 2 Vol.2 (2), 1999	Number 1 Vol.3 (1), 2000
Number 1 Vol.2 (1), 1999	Number 1 Vol.1 (1), 1998

About the journal ::: Editorial ::: Subscription ::: Information for authors ::: Contact