Anthropometric and Biochemical Measurements

The determinations of anthropometric measurements (body weight, height, neck and waist circumference), body composition by DXA (Lunar iDXA, encore 14.5, Madison, WI, USA) and blood pressure (Intelli Sense. M6, OMRON Healthcare, Hoofddorp, the Netherlands) were carried out under fasting conditions at the Metabolic Unit of the University of Navarra following standardized procedures. Blood samples were collected, processed and stored at −80 °C for further analyses [13 (link)]. Body Mass Index (BMI) was calculated as the body weight divided by the squared height (kg/m²). Biochemical determinations, including blood glucose, aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-c) and triglyceride (TG) concentrations were measured on an autoanalyzer Pentra C-200 (HORIBA ABX, Madrid, Spain) with specific commercial kits. Insulin was measured using specific ELISA kits (Demeditec; Kiel-Wellsee, Germany) in a Triturus autoanalyzer (Grifols, Barcelona, Spain). Insulin resistance was estimated using the Homeostasis Model Assessment Index (HOMA-IR), which was calculated using the formula elsewhere described [14 (link)]. The low-density lipoprotein cholesterol (LDL-c) levels were estimated using the following formula: LDL-c = TC − HDL-c − TG/5.