Biomarkers of inflammation under study were interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), fibrinogen, CRP, WBC count, and cystatin C (Cys C). Blood was collected during the in-home examination after an 8- to 10-h fast; sample processing and validation of the laboratory data have been previously reported [42 (link)]. CRP and Cys C were measured by particle-enhanced immunonephelometry using the BNII nephelometer (N High Sensitivity CRP, N Latex Cystatin C; Dade Behring) [43 (link)] and WBC count was measured using an automated analyzer (Beckman Coulter, Inc., Fullerton, CA, USA) [44 (link)] at the University of Vermont Laboratory for Clinical Biochemistry Research.
IL-6, IL-8, IL-10, and fibrinogen were measured in the cohort random sample in August 2012 at the University of Vermont Laboratory for Clinical Biochemistry Research. IL-6 was measured by ultra-sensitive ELISA (Quantikine HS Human IL-6 Immunoassay; R and D Systems, Minneapolis, MN, USA), IL-8 was measured by the Human Serum Adipokine Panel B LINCOplex Kit (Linco Research, Inc.; St. Charles, MO, USA), IL-10 was measured using the Milliplex MAP Human Cardiovascular Disease (CVD) Panel 3 (Millipore Corporation; Billerica, MA, USA) run as a single-plex assay, and fibrinogen was measured using the BNII nephelometer (N Antiserum to Human Fibrinogen; Siemens Healthcare Diagnostics, Deerfield, IL, USA).