The study also included the enrollment of healthy children with BMI appropriate for age and gender, normal liver ultrasound, and normal values for biochemical analyses. BTK inhibitor libraries They were recruited during the study period from two elementary and three middle schools in the Rome area in a pilot program to prevent cardiovascular disease (CVD) in childhood. Siblings of the study population and subjects with a history of smoking (where appropriate) or a family history of premature CVD were excluded.13 All study subjects underwent physical

examination including measurements of weight, standing height, BMI, waist circumference (WC), determination of the stage of puberty, the degree of obesity, and systolic blood pressure (BP) and diastolic BP, as reported in detail.8 The study was approved by the Hospital Ethics Committee and informed consent was obtained from subjects’ parents before assessment. Blood samples were taken after an overnight fast from each subject. Insulin, high-sensitivity C-reactive protein (CRPHS), apolipoprotein (APO) A-1 and

B were measured on a COBAS 6000 immunometric analyzer (Roche Diagnostics). Insulin concentrations were determined by an electrochemiluminescent method, CRPHS by an immunoturbidimetric method, and APO A-1 and APO B by an immunoturbidimetric method. The remaining analytes were measured on a COBAS INTEGRA 800 analyzer (Roche Diagnostics). Total NSC 683864 price cholesterol, high-density lipoprotein (HDL) cholesterol, and triglyceride concentrations were assessed by enzymatic colorimetric methods; ALT, aspartate aminotransferase (AST), and γ-glutamyl transferase

(GGT) by the enzymatic UV method; and glucose concentration by a hexokinase method. Measurements of cIMT and FMD were performed by two blinded investigators (V.C., A.M.). Longitudinal ultrasonographic scans of the carotid artery were obtained on the same day as the studies of the brachial artery reactivity and included evaluation of the right and left common carotid Thymidylate synthase arteries near the bifurcation during end diastole. We measured four values on each side and the maximum and mean cIMT were calculated. The coefficient of variation was less than 3%.8 Assessment of FMD was performed according to the guidelines of the International Brachial Artery Reactivity Task Force.5 The brachial artery was scanned above the antecubital fossa of the right arm using high-resolution vascular ultrasonography (Mylab 70 XVision Gold, 7-15-MHz linear-array transducer, Esaote, Genova, Italy). Longitudinal, electrocardiogram-gated, end-diastolic images were acquired of the brachial arterial diameter over a 1- to 2-cm segment and computer-assisted edge detection brachial analysis software was used to measure the brachial artery diameters.