Venous blood samples were analyzed for markers of low-grade inflammation (High-sensitivity C-reactive protein (hsCRP), Interleukin 6 (IL-6), Interleukin 8 (IL-8), Interleukin 10 (IL-10)) and endothelial activation (Intercellular adhesion molecule (ICAM), P-Selectin) [10 (link)].
Each participant provided fasting blood samples in the morning following a 30-min rest in the supine position. These samples were drawn from the cubital vein into 4.5 mL vacuum tubes containing 0.11 mol/L sodium citrate (Becton Dickinson, Vacutainer System Europe, Heidelberg, Germany). Plasma was prepared within 30 min through centrifugation at 2000× g for 20 min. It was later divided into aliquots, rapidly frozen in liquid nitrogen, and stored at –75 °C until further analysis. Plasma levels of hsCRP, ICAM-1, IL-6, IL-8, IL-10, and P-Selectin were assessed using xMAP® Technology, which utilizes magnetic beads conjugated with specific antibodies (all from R&D Systems, Minneapolis, MN, USA), on a MagPix instrument (Luminex Corporation, Austin, TX, USA) [10 (link)].
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