Triturus autoanalyzer

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.

The first spot urine was taken in the morning, and blood samples were obtained after 12 h overnight fasting. Biological specimens were stored frozen at a −80 °C according to approved protocols by trained technicians [56 (link)]. Biochemical analyses including glucose, hemoglobin A1c (HbA1c), triglyceride, HDL-c, total cholesterol, ALT, and AST were performed on fasting plasma by using specific kits according to manufacturer’s protocols [56 (link)]. The insulin was measured using specific ELISA kits in a Triturus autoanalyzer (Grifols, Barcelona, Spain). The Friedewald formula was used to calculate LDL-c and the VLDL-c [66 (link)].

Anthropometric variables (body weight, height and waist circumference) and body composition (Lunar iDXA, Encore 14.5, Madison, WI, USA) were assessed in fasting conditions at the Metabolic Unit of the University of Navarra following standardized procedures [45 (link)]. BMI was calculated as the body weight divided by the squared height (kg/m²). Blood samples were properly collected after overnight fasting of 8–10 h and processed at the Laboratory of Biochemistry of the University of Navarra Clinic (CUN, Pamplona, Spain). Blood glucose, triglycerides (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and gamma-glutamyltransferase (GGT) concentrations were determined on a Cobas 8000 autoanalyzer with specific commercial kits and following the instructions of the company (Roche Diagnostics, Basel, Switzerland). Insulin, fibroblast growth factor 21 (FGF-21), leptin and adiponectin concentrations were quantified with 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 computed as HOMA-IR = (insulin (μU/mL) × glucose (mmol/L))/22.5 [4 (link)]. The Triglycerides/Glucose index (TyG) (ln[triglycerides (mg/dL) × glucose(mg/dL)/2)]) was also calculated as a surrogate of glucose tolerance [46 (link)].