Gl364

Concentrations of glucose, cholesterol and triacylglycerol were measured spectrophotometrically using a commercially available enzymes assay kits (GL 364, Randox, Crumlin, UK, for glucose; CH 200, Randox, Crumlin, UK for cholesterol and TR 1697, Randox, Crumlin, UK for triacylglycerol). Concentrations of calcium and phosphate in serum were measured spectrophotometrically (using Arsenazo III method for calcium and UV for phosphate) with authomatic biochemical analyzer (Biosystem A15; Biosystems, Spain).

Blood glucose concentrations were determined in duplicate using a commercially available assay (GOD/PAP method, GL364, Randox, Ireland). Plasma insulin concentrations were determined using a commercially available ELISA, in line with the manufacturer's instructions (ELISA; Mercodia Ltd, Sweden). Blood glucose and plasma insulin total area under the curve (tAUC) following the standardized breakfast were calculated (GraphPad Prism 7, GraphPad Software, USA), using methods described previously (Wolever et al., 1991 (link)). Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) was calculated as an index of insulin resistance (Matthews et al., 1985 (link)). Plasma IL-6,−10,−15 and 1β concentrations were determined simultaneously with an automated SimplePlex immunoassay, using the anti-body-based Ella system (ProteinSimple, BioTechne, Oxford, UK), per the manufacturer's instructions (Aldo et al., 2016 (link)). All analytes were measured in triplicate and concentrations were determined using preloaded calibration curves. Serum BDNF concentrations were determined using commercially available methods, in accordance with the manufacturer's instructions (Quantikine ELISA, R & D Systems Europe Ltd, UK).

During day one of the main trials, capillary blood samples were taken at baseline, immediately post-exercise and 60-min post-exercise. Further capillary blood samples were taken 30-min, 60-min (2-h post-exercise) and 120-min (3-h post-exercise) following the standardised lunch. On day two of the main trials, a fasted capillary blood sample was taken. Following the consumption of the standardised breakfast, further blood samples were taken at 30 min, 60 min and 120 min.
For all samples, concentrations of blood glucose and plasma insulin were determined in duplicate using commercially available kits (glucose: GOD/PAP method, GL364, Randox, Crumlin, Ireland; insulin: ELISA, Mercodia Ltd, Uppsala, Sweden). Blood glucose and plasma insulin tAUC following the standardised lunch on day one and the standardised breakfast on day two were calculated using previously described methods [19 (link)]. HOMA-IR was calculated as fasted insulin (mU·L) × fasting glucose (nmol·L)/22.5 [20 (link)].