Balkan Journal of Medical Genetics

LACK OF ASSOCIATION BETWEEN VARIANTS WITHIN THE AHSG, HCRT AND NPY2R GENES AND ANTHROPOMETRICAL PARAMETERS IN CZECH POST-MONICA STUDY
Jurcikova L*, Adamkova V, Lanska V, Suchanek P, Hubacek JA
*Corresponding Author: Ing. Lucie Jurčíková, Institute for Clinical and Experimental Medicine, DEM, Videnska 1958/9, Prague 4, 14021, Czech Republic; Tel.: +420-261-362-229; Fax: +420-241-721-574; E-mail: jurl@ikem.cz
page: 63

MATERIALS AND METHODS

We analyzed a randomly selected representative sample of adult individuals (1191 males and 1368 females), aged 25-64 years at the time of first examination (Table 1). All individuals had participated in the Czech post-MONICA study, performed according to the World Health Organization MONICA Project protocol, which was aligned to examine risk factors of cardiovascular disease development, including BMI and plasma lipids, after overnight fasting [24,25]. The subjects were examined in nine Czech districts (Kromeriz, Chrudim, Cheb, Jindrichuv Hradec, Pardubice, Litomerice, Plzen, Prague East, and Benesov) in 1997/1998 and were reexamined in 2000/ 2001 and 2007/2008. For our purposes, the most recent data (2007/2008) were used. Written informed consent was given by all individuals. The study was approved by the Institutional Ethics Committee and is in agreement with the Helsinki Declaration of 1975. Polymorphisms were genotyped using the the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (for more details see Table 2) in a total volume of 25 mL. All PCR chemicals and restriction enzymes used were from Fermentas International Inc., Burlington, Ontario, Canada. The PCR reactions were performed on the PCR cyclers MJ Research DYAD Disciple. Restriction fragments were separated via horizontal electrophoresis on 10.0% polyacrylamide gel using the MADGE platform. To check our PCR-RFLP results, we repeated 5.0% of the samples in each polymorphism. The results of SNPs in the AHSG, HCRT and NPY2R genes were identical in 100.0% of cases. Hardy-Weinberg equilibrium was proved by the c2 test to confirm the independent segregation of the alleles. Differences between genotypes were calculated by analysis of variance (ANOVA), logarithmic transformation was used for triglyceride levels. Mean values ± SD were calculated, two-tailed p values of less than 0.05 were considered to be significant (Table 3).

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