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All parents gave their written consent, and children gave their verbal and written assent. Anthropometric Measurements and Blood Pressure Weight was measured to the nearest 0.1 kg and height to the nearest 0.1 cm. BMI was calculated as weight (kg)/height (m2); percentage body fat was assessed by bioelectrical impedance23 and body surface area (BSA) was calculated with Haycock formula.24 Blood pressure was measured with a mercury sphygmomanometer after 20 minutes rest, in a supine position. Three sizes of cuff were used (9��21, 11��36, and 12��41 cm); the cuff width was required to cover two-thirds of the length of the [http://en.wikipedia.org/wiki/Laccase Laccase] child��s arm. Blood Sampling Blood samples of all children were collected after 12 hours fasting from a vein in the antecubital fossa, without venous occlusion. Before collecting the blood, we asked parents of each child the hour of last food for fasting confirmation. Blood samples were separated into aliquots and frozen immediately at ?70��C until analysis to avoid interassay variability. Glucose, UA, cholesterol, triglycerides (TG), and high density lipoprotein (HDL), and LDL concentrations were measured using an automatic analyzer (Roche Diagnostics, Mannheim, Germany). Blood count was determined by hematologic analyzer (Nihon Kohdem Corporation, Japan). Leptin and sP-selectin were determined by an enzyme immunoassay (Invitrogen Corporation, CA, USA). vWF:Ag was determined by enzyme-linked immunosorbent assay (ELISA) (Immubind, American Diagnostica, USA). PAI-1 was determined by ELISA (Prepro-Tech Inc, [http://www.selleckchem.com/TGF-beta.html TGF-beta inhibitor] [http://www.selleckchem.com/products/Thiazovivin.html Thiazovivin supplier] USA). In all determinations, the intra- and interassay variability was  samples. Pearson��s correlation and linear regression coefficients were used to analyze the relation between variables. The independence of association of leptin with platelet activation was assessed by logistic regression analysis (when dependent variable, ie. platelet activation [sP-selectin] was entered as a categorical variable stratifying into two groups according to the 75th percentile observed (44.32 ng/mL). Univariate linear regression analyses and stepwise regression models were used to test the predictive association of biochemical parameters (Leptin, PAI-1, vWF, UA, glucose, cholesterol, triglycerides, LDL, HDL, and PAI-1) with platelet activation. A P