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Shortened leukocyte telomere length (LTL), a purported marker of cellular aging, is associated with morbidity and mortality. However, the association of physical activity, a modifiable lifestyle behavior, with LTL has not been adequately studied among older adults.In this cross-sectional study, we examined associations of various intensity levels of leisure-time physical activity with LTL among 1476 older white and African American women from the Women's Health Initiative Objective Physical Activity and Cardiovascular Health study. Self-reported physical activity was assessed by questionnaire, and LTL was measured by Southern blot. The association between physical activity and LTL was evaluated using multiple linear regression models adjusted for demographic characteristics, lifestyle behaviors, and health-related variables.Women were on average aged 79.2 (standard deviation 6.7) years old. In the final model adjusted for age, race/ethnicity, education, marital status, smoking, alcohol, body mass index, a history of chronic diseases, and hormone therapy use, LTL was on average 110 (95% confidence interval, 20–190) base pairs longer among women in the highest (≥17.00 MET-hours/week) compared with the lowest (<1.25 MET-hours/week) level of total leisure-time physical activity (P for trend = 0.02). Higher levels of moderate-to-vigorous physical activity (P for trend = 0.04) and faster walking speed (P for trend = 0.03) were also associated with longer LTL in the fully-adjusted models.Older women participating in greater amounts of total leisure-time physical activity and moderate-to-vigorous physical activity had longer LTL.A cross-sectional of physical activity and LTL in older women was performed.Greater total leisure-time physical activity was associated with longer LTL.Greater moderate-to-vigorous physical activity was associated with longer LTL.Faster walking speed was associated with longer LTL.