2.4. Analysis of Blood Samples
BLOOD, ADHESION
At baseline and week 4, participant blood samples were collected from the antecubital vein via standard phlebotomy techniques into appropriate vacutainers. Blood was collected in 4 mL dipotassium EDTA tubes and 8.5 mL serum separator tubes. After centrifugation, a portion of collected serum was aliquoted and stored at −80 °C until analyses of cytokines and adhesion molecules. Whole blood was transported and analyzed by Sonora Quest Laboratories (Arizona) for complete blood count with differentials, which was utilized for computation of neutrophil/lymphocyte ratios (NLR). Serum was also analyzed for high sensitivity C-reactive protein (hsCRP) and ferritin by Sonora Quest Laboratories. Soluble P-selectin (sP-selectin) and soluble intercellular adhesion molecule 1 (sICAM-1) levels in serum samples were analyzed via ELISA methods (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). Circulating serum cytokines and chemokines were analyzed via human-focused 15-plex discovery assay (Eve Technologies Corporation, Calgary, Canada). This biomarker panel provided quantitation of IL-6, IL1-β, IL-1receptor agonist (Ra), IL-2, IL-4, IL-5, IL-8, IL-10, IL-12p40, IL-12p70, IL-13, MCP-1, IFN-γ, granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-α. 