Blood samples were collected from the antecubital vein under fasting conditions and stored at -80° C until use. Blood count, lipid profile (TC, LDL-c, HDL-c, and TG), serum glucose, and renal and hepatic function were assessed using standard laboratory techniques. Lp(a) levels were evaluated using a standard immunochemical-based assay with reference ranges < 30 mg/dl or <75 nmol/l [16 , 17 (link)].
The LDL peak buoyancy and cholesterol distribution across the lipoprotein classes were assessed by density gradient ultracentrifugation (DGUC) for apo B containing lipoproteins, optimized for LDL particles. Thirty-seven 0.45 ml fractions were then collected from the bottom of the centrifuge tube. Cholesterol was measured in each fraction. The relative flotation rate (Rf), which characterizes LDL peak buoyancy, was obtained by dividing the fraction number containing the LDL-cholesterol peak by the total number of fractions collected. Each lipoprotein subclass elution range was defined as previously published [18 (link)]. Circulating levels of osteoprotegerin (OPG) were assessed by using dedicated ELISA kits (R&D System), whereas serum levels of high-sensitivity CRP (hs-CRP) have been measured through a sensitive immunonephelometric method (Dade Behring).
The LDL peak buoyancy and cholesterol distribution across the lipoprotein classes were assessed by density gradient ultracentrifugation (DGUC) for apo B containing lipoproteins, optimized for LDL particles. Thirty-seven 0.45 ml fractions were then collected from the bottom of the centrifuge tube. Cholesterol was measured in each fraction. The relative flotation rate (Rf), which characterizes LDL peak buoyancy, was obtained by dividing the fraction number containing the LDL-cholesterol peak by the total number of fractions collected. Each lipoprotein subclass elution range was defined as previously published [18 (link)]. Circulating levels of osteoprotegerin (OPG) were assessed by using dedicated ELISA kits (R&D System), whereas serum levels of high-sensitivity CRP (hs-CRP) have been measured through a sensitive immunonephelometric method (Dade Behring).