Blood Sampling and Adipokines Measurements

Blood samples were collected at pre- and post-intervention from an antecubital vein in sitting position. Participants were required to meet the following criteria for blood sampling: (1) no exercise other than the prescribed exercise regime of the study at least 72 h before blood sampling, and (2) No drink or consume specific beverage or food such as coffee, dark tea, banana, and cereal 24 h before blood sampling. Blood samples were placed into EDTA (plasma) and sterile (serum) tubes. All samples were centrifuged and then were stored at −70°C until analysis. Following adipokines were measured using commercially available assay kits: (1) plasma visfatin (Cat. No. V0523EK, enzyme-linked immunosorbent [ELISA] assay kit, AdipoGen, Seoul, Korea); (2) serum omentin-1 (Cat. No. APO-54N-034, sandwich ELISA, ELISA kit, Apotech Corp., Switzerland); (3) plasma vaspin (Cat. No. CSB-E09771h, ELISA kit, Cusabio Biotech, Wuhan, China); (4) plasma FGF-21 (Cat. No. RD191108200R, sandwich ELISA kit, Biovendor, Heidelberg, Germany); (5) plasma adiponectin (Cat. No. RD191023100, sandwich ELISA kit, Biovendor, Heidelberg, Germany); (6) plasma leptin (Cat. No. RD191001100, sandwich ELISA kit, Biovendor, Heidelberg, Germany); and (7) plasma ghrelin (Cat. No. 171B7104M, ELISA kit - Bio-Plex 200 System, Bio-Rad Laboratories, Gurgaon, India). The Elisa machine used in the current project was made in Biohit Company in Finland. It was programmed to CV = 7.59 in terms of qualitative control. Moreover, in order to boost the measurement accuracy; all the tests were double checked.