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Sleep and sedentary and active behaviors are linked to cardiovascular disease risk biomarkers, and across a 24-hour day, increasing time in 1 behavior requires decreasing time in another. We explored associations of reallocating time to sleep, sedentary behavior, or active behaviors with biomarkers. Data (n = 2,185 full sample; n = 923 fasting subanalyses) from the cross-sectional 2005–2006 US National Health and Nutrition Examination Survey were analyzed. The amounts of time spent in sedentary behavior, light-intensity activity, and moderate-to-vigorous physical activity (MVPA) were derived from ActiGraph accelerometry (ActiGraph LLC, Pensacola, Florida), and respondents reported their sleep duration. Isotemporal substitution modeling indicated that, independent of potential confounders and time spent in other activities, beneficial associations (P < 0.05) with cardiovascular disease risk biomarkers …