Comprehensive Lipid and Inflammation Profile

Lipid profile, C-reactive protein (CRP), total apoJ, Lp-PLA₂ activity, LDL size, and HDL subfraction proportion were determined in plasma obtained in EDTA-containing Vacutainer tubes. The lipid profile included total cholesterol (Roche), triglycerides (Roche), apoB (Roche), and VLDL, LDL, and HDL cholesterol. Cholesterol of lipoprotein fractions was quantified using a direct HDL-cholesterol method (HDL-C plus, Roche) or by ultracentrifugation when the TG concentration was higher than 3 mmol/L, according to the National Cholesterol Education Program [25 (link)]. Non-esterified fatty acids (NEFA) were quantified using a commercial kit (Wako Pure Chemical, Osaka, Japan). All these determinations and CRP (Roche Diagnostics, Basel, Switzerland) were performed in a Cobas 6000/c501 autoanalyzer. ApoJ plasma levels were measured with commercial ELISA kits (Mabtech, Stockholm, Sweden). LDL size and HDL subfraction proportions were evaluated from plasma by non-denaturing polyacrylamide gradient (2.5%–16%) gel electrophoresis, as described previously [26 (link)]. Lp-PLA₂ activity was determined using 2-thio-PAF (Cayman Chemicals, Ann Arbor, MI, USA) as a substrate [27 (link)] according to the manufacturer’s instructions. The distribution of Lp-PLA₂ between lipoprotein fractions was measured by precipitating apoB-containing lipoproteins from plasma with dextran sulfate [28 (link)].