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This study describes the developmental changes and individual stability in autonomic nervous system (ANS) resting and challenge responses for a cohort of primarily Latino, low-income children during the first 5 years of life.ANS measures of the parasympathetic nervous system (respiratory sinus arrhythmia [RSA]) and sympathetic nervous system (preejection period [PEP]) were collected on a representative sample of the full cohort at 6, 12, 42, and 60 months of age (N = 378). The children participated in a standardized protocol to elicit ANS responses during resting and challenging states. Reactivity profiles were created to summarize each child's combined RSA and PEP reactivity (i.e., change in response to challenges compared to a resting state).Results showed developmental changes in ANS measures from 6 to 60 months: heart rate decreased, RSA increased, PEP increased, and frequency of classic reactivity profiles of reciprocal sympathetic activation and parasympathetic withdrawal increased. Correlations showed moderate stability for resting and challenging conditions but not reactivity.These findings suggest that low-income Latino children, from 6 to 60 months of age, showed ANS developmental changes and moderate individual stability for resting and challenge responses but not for reactivity. There was a significant shift in the frequency of children with the classic reactivity profile from 6 by 60 months of age. This is the first cohort study to show the developmental changes in ANS and young children's increase in their biologic sensitivity to the environment during the first 5 years of life.