Circulating Biomarkers
Morning fasting blood samples were collected between 8 and 10 AM at baseline and at treatment completion. Serum aliquots for insulin and high-sensitive C-reactive protein (CRP) were measured on frozen samples stored at −80°C until assayed, whereas glucose and total cholesterol were measured on fresh samples. Insulin was measured with an electrochemiluminescence immunoassay by Cobas e411 (Roche Diagnostics, Mannheim, Germany). Serum concentrations of glucose, total cholesterol, and CRP were determined by COBAS INTEGRA 800 (Roche Diagnostics). The HOMA index more than 2.8 was the cutoff of insulin resistance based on a population study conducted in Italy.24