Modular 700

After peripheral blood sampling, plasma glucose and serum triglyceride levels were measured automatically (Modular 700, Roche, Meylan, France). Serum insulin was measured using a radio-immunometric sandwich assay (CIS bio international, Gif-Sur-Yvette, France).

Heparinized plasma glucose, triglycerides, total and HDL-cholesterol concentrations in samples were determined using enzymatic methods and spectrophotometry (Modular 700, Roche Diagnostics, Meylan France). Low-density lipoprotein (LDL)-cholesterol was calculated by Fried-wald’s formula [Total cholesterol–(HDL cholesterol + triglycerides/5)]. Serum insulin was measured by immunoradiometric sandwich assay (Bis-Insulin IRMA^®, CisBio international, Gif-Sur-Yvette, France). The homeostasis model assessment resistance (HOMA-RI) index was calculated by the following equation: insulin (μIU/mL) x glucose (mmol/L)/22.5. Serum high-sensitive C-reactive protein (hsCRP) level was measured using automated immunonephelometry (Behring Nephelometer II Analyzer^®, Dade Behring, Germany). Leptin and TNFα were measured by commercially available multiplex beads immunoassays (Fluorokine MAP Multiplex Human Cytokine Panel and Obesity Panel, R&D Systems, Minneapolis, USA) and read by the Bioplex 200 array reader (Bio-Rad Laboratories, Hercules, CA, U.S.A.) which uses Luminex xMAPTM Technology (Luminex Corporation, Austin, TX, U.S.A.).