Blood samples were taken between 8:00 and 9:00 a.m. after overnight fasting. Fasting plasma glucose (FPG), hemoglobin A1c (HbA1c), insulin, lipid profile (total cholesterol, high-density lipoprotein (HDL) cholesterol, and triglycerides), uric acid, and liver markers were analyzed in serum. Using duplicate samples, the radioimmunoassay method (Behring, Scoppito, Italy) was applied to measure the serum insulin concentrations. A competitive luminometric method based on the solid-phase antigen luminescent technology (SPALT) principle was employed to assess the serum concentrations of TSH, FT3, and FT4 (LIAISON FT3, FT4, TSH, DiaSorin, Saluggia, Italy). Fasting plasma lipid concentrations (triglycerides, total cholesterol, and HDL cholesterol) were measured using an automated colorimetric method, and fasting plasma glucose concentrations were determined using the glucose oxidase method (Sclavus, Siena, Italy) (Hitachi; Boehringer Mannheim, Mannheim, Germany). An Architect c8000 chemical analyzer was used to measure glycated hemoglobin (HbA1c) (Abbott).
The URICASE/POD method was employed to quantify the amount of uric acid in the blood using an autoanalyzer (Boehringer Mannheim). Standard laboratory procedures were used to measure amino transferase, -glutamyl transpeptidase (GT), and creatinine with an automated system (UniCel Integrated Workstations DxC 660i, Beckman Coulter, Fullerton, CA, USA). The Friedewald equation [30 (link)] was used to calculate LDL cholesterol. DxI/Access was used to perform a quantitative analysis of serum ferritin using Access Ferritin Reagent Packs (Beckman-Coulter AB, Bromma, Sweden). Radioimmunoassay (Behring, Scoppito, Italy) was used to measure the levels of serum insulin, and chemiluminescence was applied to determine the levels of serum 25(OH) vitamin D (Diasorin Inc., Stillwater, OK, USA). Insulin resistance was calculated using the Homeostasis Model Assessment-Insulin Resistance (HOMA-IR) method: ((fasting insulin × fasting glucose)/405, normal range 0.23–2.5) [31 (link)].
The URICASE/POD method was employed to quantify the amount of uric acid in the blood using an autoanalyzer (Boehringer Mannheim). Standard laboratory procedures were used to measure amino transferase, -glutamyl transpeptidase (GT), and creatinine with an automated system (UniCel Integrated Workstations DxC 660i, Beckman Coulter, Fullerton, CA, USA). The Friedewald equation [30 (link)] was used to calculate LDL cholesterol. DxI/Access was used to perform a quantitative analysis of serum ferritin using Access Ferritin Reagent Packs (Beckman-Coulter AB, Bromma, Sweden). Radioimmunoassay (Behring, Scoppito, Italy) was used to measure the levels of serum insulin, and chemiluminescence was applied to determine the levels of serum 25(OH) vitamin D (Diasorin Inc., Stillwater, OK, USA). Insulin resistance was calculated using the Homeostasis Model Assessment-Insulin Resistance (HOMA-IR) method: ((fasting insulin × fasting glucose)/405, normal range 0.23–2.5) [31 (link)].
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