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This study examined the relation of motivational systems to patterns of autonomic responses to stress. Specifically, we examined patterns of physiological response resulting from differential activation of motivational systems for behavioral approach and behavioral inhibition. We also examined the relation of these motivational systems to threat and challenge responses to potential stress. Self-report, cardiac measures (i.e., pre-injection period, or PEP, and heart rate, or HR), and blood pressure (i.e., systolic blood pressure, or SBP, and diastolic blood pressure, or DBP) measures reliably distinguished between an experimental condition facilitating behavioral approach and an experimental condition facilitating coactivation of behavioral approach and behavioral inhibition systems. Vascular (e.g., total peripheral resistance, or TPR) and electrodermal (e.g., electrodermal response, or EDR) measures, however, did not vary by condition. We discuss the results in relation to other research on motivational factors as they relate to stress and cardiovascular reactivity....