2.4. Circulating Biomarkers Assessment
TNF-α
BLOOD
Fasting blood samples were obtained at baseline and the end of the study after 4–6 weeks of intervention. Blood samples were processed into serum and aliquots were frozen at −80 °C until biomarkers measurements.We measured hs-CRP, glycemia, and insulin using the Abbott Alinity analyzer (Abbott Diagnostics, Abbott Park, IL, USA). We then calculated the HOMA-IR index as an indicator of metabolic status [Total IGF-I, IGFBP-3, and 25OHD concentrations were assessed by chemiluminescent immunometric assay on the IDS-iSYS analyzer (Immunodiagnostic Systems Limited, East Boldon, UK).Adiponectin, leptin, IL-10, IL-6, and TNF-α were measured on the automated platform ELLA (ProteinSimple, BioTechne, Minneapolis, MN, USA), as previously described [Samples from the same participant were analyzed in the same batch. The laboratory staff were blinded to the arm assignment.