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Amplification and digestion were carried out by triplicate in all cases. Restriction fragments of ADH1B characterizing each genotype were as follows: a nondigested fragment of 161?bp for *1/*1 genotype; 161, 98, and 63?bp for *1/*2 genotype; 98 and 63?bp for *2/*2 genotype. For ALDH2, 112 and 23?bp corresponded to *1/*1 genotype; 135, 112, and 23?bp to *1/*2 genotype; and selleck compound a nondigested fragment of 135?bp to *2/*2 genotype. CYP2E1 exhibited 143 and 423?bp for the common genotype c1/c1; 566, 423, and 143?bp for the heterozygous genotype c1/c2; and a nondigested fragment of 566?bp for the polymorphic genotype c2/c2. Samples were taken in a fasting state of 12?hours. Serum determination of total cholesterol, cholesterol bound to high density lipoproteins (HDL-c), cholesterol bound to low density lipoproteins (LDL-c), cholesterol bound to very low density lipoproteins (VLDL-c), triglycerides, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TB), and albumin were performed using commercially available kits (Merck Co., Whitehouse Station, NJ). Genotype and allele frequency differences between groups were assessed through the Pearson chi-squared test or Fisher��s exact test when n values lesser than 10. Quantitative parameters were compared through ANOVA. Probability values less than 0.050 were considered GRB10 statistically significant. Written informed consent was obtained from all subjects and the study protocol conformed to the ethical guidelines according to the 1975 Declaration of Helsinki. We studied 101 healthy HUI, 65 women and 41 men, aged 20 to 78?years from this website the HUI community living in the Indigenous reservation ��Jes��s Mar��a,�� located at the Nayar region of Nayarit state at the Sierra Madre Occidental; and 331 healthy subjects, 193 women and 138 men, aged 31 to 75?years, from a MES population living in the city of Tepic, Nayarit, in western Mexico. The PCR amplification protocol for ADH1B, ALDH2, and CYP2E1 utilized in this study provided an efficient reproducible time saving alternative that can be widely recommended, since in a separate tube but in the same running of PCR the 3 polymorphic segments can be obtained. Restriction patterns according to genotype were observed (Fig.?1). Genotype distribution and allele frequency of ADH1B, ALDH2, and CYP2E1 polymorphisms are shown in Table?1. Similar distribution of ADH1B genotypes were observed both in HUI and MES individuals, although HUI subjects were monomorphic for ADH1B*1/1 genotype. Meanwhile, frequency of the ADH1B*2 polymorphic allele in MES (3.4%) attained statistical significance (p?=?0.008) when compared with HUI (0.0%). Conversely, MES were monomorphic for ALDH2*1/1 genotype and polymorphic allele frequency in HUI (0.5%) and MES (0.0%) did not show statistically significant difference. Regarding CYP2E1, significantly higher (p?