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Background  A recent paradigm shift has highlighted the importance of considering how sleep, physical activity and sedentary behaviour work together to influence health, rather than examining each behaviour individually. We aimed to determine how adherence to 24-h movement behavior guidelines from infancy to the preschool years influences mental health and self-regulation at 5 years of age.    Methods  Twenty-four hour movement behaviors were measured by 7-day actigraphy (physical activity, sleep) or questionnaires (screen time) in 528 children at 1, 2, 3.5, and 5 years of age and compared to mental health (anxiety, depression), adaptive skills (resilience), self-regulation (attentional problems, hyperactivity, emotional self-control, executive functioning), and inhibitory control (Statue, Head-Toes-Knees-Shoulders task) outcomes at 5 years of age. Adjusted standardised mean differences (95% CI) were determined between those who did and did not achieve guidelines at each age.    Results  Children who met physical activity guidelines at 1 year of age (38.7%) had lower depression (mean difference [MD]: -0.28; 95% CI: -0.51, -0.06) and anxiety (MD: -0.23; 95% CI: -0.47, 0.00) scores than those who did not. At the same age, sleeping for 11–14 h or having consistent wake and sleep times was associated with lower anxiety (MD: -0.34; 95% CI: -0.66, -0.02) and higher resilience (MD: 0.35; 95% CI: 0.03, 0.68) scores respectively. No significant relationships were observed at any other age or for any measure of self-regulation. Children who consistently met screen time guidelines had lower anxiety (MD: -0.43; 95% CI: -0.68, -0.18) and depression (MD: -0.36; 95% CI: -0.62, -0.09) scores at 5. However, few significant relationships were observed for adherence to all three guidelines; anxiety scores were lower (MD: -0.42; 95% CI: -0.72, -0.12) in the 20.2% who adhered at 1 year of age, and depression scores were lower (MD: -0.25; 95% CI: -0.48, -0.02) in the 36.7% who adhered at 5 years of age compared with children who did not meet all three guidelines.    Conclusions  Although adherence to some individual movement guidelines at certain ages throughout early childhood was associated with improved mental health and wellbeing at 5 years of age, particularly reduced anxiety and depression scores, there was little consistency in these relationships. Future work should consider a compositional approach to 24-h time use and how it may influence mental wellbeing.    Trial registration  ClinicalTrials.gov number  NCT00892983

Adherence to 24-h movement behavior guidelines and psychosocial functioning in young children: a longitudinal analysis