The patients’ adipokine levels (chemerin, leptin, adiponectin, resistin and visfatin) were assessed in serum obtained after > 12 h of fasting. Serum adipokine concentrations were quantified by ELISA (Quantikine Human Immunoassay; R&D Systems, Minneapolis, MN, USA). Leptin sensitivity was 7.8 pg/mL, adiponectin sensitivity was 0.08 ng/mL, chemerin sensitivity was 1.08 pg/mL, resistin sensitivity was 0.01 ng/mL, and visfatin sensitivity was 1 ng/mL (Mybiosource San Diego California USA).

Additionally, serum pro-inflammatory cytokines (TNF-α, IL-6, IL-1β and IL-18) were measured by ELISA using commercial kits. TNF-α sensitivity was 0.5 pg/mL, IL-6 sensitivity was 0.7 pg/mL, and IL-1β sensitivity was 1 pg/mL (R&D Systems, Minneapolis, MN, USA). For IL-18, the sensitivity was 12.5 pg/mL (MBL International).

The assay for the adhesion molecule E-selectin had a sensitivity of 0.009 ng/mL (R&D Systems, Minneapolis MN USA). All procedures for the measurement of these molecules were conducted by researchers blinded to the patients’ clinical characteristics.

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