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Objective Although the relationship between religious practice and health is well established, the relationship between spirituality and health is not as well studied. The objective of this study was to ascertain whether participation in the mindfulness-based stress reduction (MBSR) program was associated with increases in mindfulness and spirituality, and to examine the associations between mindfulness, spirituality, and medical and psychological symptoms. Methods Forty-four participants in the University of Massachusetts Medical School's MBSR program were assessed preprogram and postprogram on trait (Mindful Attention and Awareness Scale) and state (Toronto Mindfulness Scale) mindfulness, spirituality (Functional Assessment of Chronic Illness Therapy—Spiritual Well-Being Scale), psychological distress, and reported medical symptoms. Participants also kept a log of daily home mindfulness practice. Mean changes in scores were computed, and relationships between changes in variables were examined using mixed-model linear regression. Results There were significant improvements in spirituality, state and trait mindfulness, psychological distress, and reported medical symptoms. Increases in both state and trait mindfulness were associated with increases in spirituality. Increases in trait mindfulness and spirituality were associated with decreases in psychological distress and reported medical symptoms. Changes in both trait and state mindfulness were independently associated with changes in spirituality, but only changes in trait mindfulness and spirituality were associated with reductions in psychological distress and reported medical symptoms. No association was found between outcomes and home mindfulness practice. Conclusions Participation in the MBSR program appears to be associated with improvements in trait and state mindfulness, psychological distress, and medical symptoms. Improvements in trait mindfulness and spirituality appear, in turn, to be associated with improvements in psychological and medical symptoms.