Objective: Inconsistent findings reported for the effect of physical activity on lipids, lipoproteins, and blood pressure in children may be due to errors inherent in the methods used to measure physical activity, lack of control for other cardiovascular risk factors, or both. The purpose of this study was to evaluate the association between physical activity assessed using direct measures of energy expenditure and cardiovascular risk factors, controlling for dietary intake and percent body fat., Methods: Nonresting energy expenditure was determined in 49 8- to 11-year-old girls from measurements of daily energy expenditure (using doubly-labeled water, 2H2(18)O) and resting metabolic rate (using indirect calorimetry). Self-reported recall of the hours of participation in physical activities of at least moderate intensity (energy expenditure at least four times the resting metabolic rate, METS > or = 4) during the previous year was also obtained. Percent body fat was estimated from the measurement of total body water with H2(18)O. Concentrations of total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol (LDL-C), apolipoprotein B (apo B), apo A-I, lipoprotein (a), insulin, and estradiol, as well as the waist-to-hip ratio, systolic blood pressure, diastolic blood pressure, and dietary intake from 7-day food records were measured. Data were analyzed using Pearson product-moment correlation and stepwise multiple regression., Results: Self-reported hours of participation in activities with METS (metabolic equivalents) of 4 or greater significantly predicted LDL-C and apo B concentrations, even after adjustment for percent body fat and percentage of dietary energy from saturated fat. Nonresting energy expenditure adjusted for weight, a measure of the energy spent on physical activity, did not predict LDL-C or high-density lipoprotein cholesterol concentrations. Body mass index and insulin concentration predicted systolic and diastolic blood pressure, respectively., Conclusions: These findings suggest that the intensity of physical activity may be a more important determinant of LDL-C in children than the energy spent on physical activity.