2.4. Metabolic Biomarkers
high-density lipoprotein, TG, TNF-α
STASIS
Venous blood samples were collected into one tube between 7:00 am and 9:00 am after a 12 h fast and at least 48 h since the last RT session. Five milliliters were withdrawn from a prominent superficial vein in the antecubital space using a clean venous puncture with minimal stasis and placed in a tube containing a dipotassiumethylenediaminetetra-acetic acid (EDTA) as an anticoagulant and preservative. All samples were centrifuged at 3000 rpm for 15 min, and plasma or serum aliquots were stored at −80 °C until assayed. Inter- and intra-assays coefficients of variation were <10% as determined in human plasma. Glucose, total cholesterol (TC), high-density lipoprotein (HDL-c), triglycerides (TG), and high-sensitivity C-reactive protein concentrations were determined by standard methods in a specialized laboratory at University Hospital. The LDL-c was calculated using the following equation: LDL-c = TC − (HDL-c) − (TG/5), with TG/5 = VLDL-c. The analyses were performed using a biochemical auto-analyzer system Dimension RxL Max (Siemens Healthcare Diagnostics, Malvern, PA, USA) according to established methods in the literature consistent with the manufacturer’s protocol. Interleukin-10 (IL-10), Interleukin-6 (IL-6), and TNF-α were determined by enzyme-linked immunosorbent assay (ELISA), according to the specifications of the manufacturer (R&D Systems, Minneapolis, MN, USA). Advanced oxidation protein products (AOPP) were determined in the plasma using a semiautomatic method described by Witko-Sarsat et al. [