Variation in the APOE, lipoprotein lipase (LPL) and cholesterol ester transfer protein (CETP) genes has been consistently associated with variation in lipid levels in adults [6] and [7]. However, genetic variation in these gene loci explain only a modest proportion of inter-individual variability in fasting lipid levels [8]. We

genotyped the GENDAI cohort for ten variants in the APOE, LPL, CETP genes and the APOA5/A4/C3 cluster, to examine if the reported Screening Library in vitro effects could be replicated in children and assess if these associations could be further modulated by body mass index (BMI). Participants were drawn from the children recruited in the GENDAI study. Briefly, a random sample of 2492 children attending school in the Attica region in Greece were invited to join the study. A total of 1138 children were recruited from November 2005 to June 2006. Due to the heterogeneity in allele frequencies between Greek and non-Greek Caucasians, only children of Greek nationality, mean age: 11.2 ± 0.7 years (n = 882; 418 males and 464 females), were included in the present study. Details of recruitment and data collection have been previously described [3].

All persons gave their informed consent prior to inclusion in the study. The study was approved by the Institutional Review Board of Harokopio University and the Greek Ministry learn more of Education [3]. A salting-out procedure [9] was used to isolate DNA samples from whole blood. Ten single nucleotide polymorphisms (SNPs) in six candidate genes; LPL S447X (rs328), CETP Taq1B (rs708272), APOE (rs7412, rs429358), APOA5 −1131C > T (rs662799) and S19W (rs3135506), APOA4 S347T (rs675) and APOC3 −482C > T (rs2854117), 1100C > T (rs4520) and 3238C > G (rs5128) were genotyped using TaqMan technology

(Applied Biosciences, ABI, Warrington UK). Reactions were performed on 384-well microplates and analysed using ABI TaqMan 7900HT software. Primers and MGB probes are available upon request. Hardy–Weinberg equilibrium (HWE) CYTH4 was examined by chi-square goodness of fit test. A p value of <0.05 was taken as deviation from HWE. Plasma levels of insulin, TG, total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) and all anthropometric measures were natural log-transformed. For association studies a p value of <0.01 was taken as statistically significant. Setting a threshold of significance was the chosen method above Bonferroni corrections, since the candidate genes studied had been selected for based on a priori hypothesis and biological plausibility. A p value of <0.05 was taken as statistically significant in Principal Component Analysis (PCA). The majority of statistical analyses were performed using Intercooled Stata 8.2 for Windows (StataCorp LP, Texas, USA). Haplotype association analysis was carried out using Thesias [10]. PCA was carried out using SAS (SAS Institute Inc., Cary, NC).