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This study was conducted to determine the effect of physical education class enrollment and physical activity on academic achievement in middle school children.Participants were 214 sixth-grade students randomly assigned to physical education during either first or second semesters. Moderate and vigorous physical activity (MVPA) (number of 30-min time blocks) outside of school was assessed using the 3-d physical activity recall (3DPAR). The 3DPAR time blocks were converted to ordinal data with scores of 1 (no activity), 2 (some activity), or 3 (activity meeting Healthy People 2010 guidelines). Academic achievement was assessed using grades from four core academic classes and standardized test scores (Terra Nova percentiles).Grades were similar regardless of whether students were enrolled in physical education during first or second semesters. Physical education classes averaged only 19 min of MVPA. Students who either performed some or met Healthy People 2010 guidelines for vigorous activity had significantly higher grades (P < 0.05) than students who performed no vigorous activity in both semesters. Moderate physical activity did not affect grades. Standardized test scores were not significantly related to physical education class enrollment or physical activity levels.Although academic achievement was not significantly related to physical education enrollment, higher grades were associated with vigorous physical activity, particularly activity meeting recommended Healthy People 2010 levels.