2.4. Measurements
tumor necrosis
BLOOD, TUMOR NECROSIS
Blood samples were collected at the beginning and end of each of the three breakfast interventions, following an overnight fast. Blood was drawn from the antecubital vein into 9 mL tubes. Following venipuncture, the samples were kept on ice and subsequently centrifuged at 10,000× Cytokines (interferon (IFN)-γ, interleukin (IL)-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor (TNF)-α), as well as growth factor vascular/endothelial growth factor (VEGF) and epidermal growth factor (EGF) were analyzed using an Evidence Investigator TM Cytokine I High-Sensitivity Array (following the protocol provided by Randox Laboratories Ltd., Crumlin, UK) utilizing a sandwich immunoassay technique. In brief, 25 µL of serum samples were applied to the biochip and incubated overnight under stable temperature conditions in a thermoshaker. After incubation, the biochips were manually inserted into the image station, and the light signal generated by each analyte was simultaneously detected using digital imaging software provided by the manufacturer. The sensitivity of each analyte, measured by the intra-assay coefficient of variation, was as follows: EGF: 4.6%, IFNγ: 12.7%, IL-1α: 9.7%, IL-1β: 8.2%, IL-2: 9.6%, IL-4: 10.7%, IL-6: 12.9%, IL-8: 10.1%, IL-10: 6.3%, MCP-1: 6.8%, TNFα: 10.2%, and VEGF: 7.3%. Additionally, C-reactive protein (CRP) was determined using an immunoassay with an Invitrogen