Background: Rotating and night-including shifts disrupt circadian alignment, impair sleep, and may reduce nurses’ physiological recovery. Objectives: This study aimed (1) to compare sleep quality and physical well-being across four shift schedules among hospital nurses; and (2) to examine whether the association between rotating shifts and physical well-being was statistically consistent with an indirect association via sleep quality. Methods: In this cross-sectional study, 173 nurses from a ter-tiary hospital in Zagreb, Croatia, completed validated measures of sleep quality and physical well-being. Four shift patterns were analyzed—fixed morning, morning–afternoon, extended 12-h, and rotating three-shift—using Welch ANOVA and regression models. A bootstrapped mediation analysis (10,000 resamples; BCa method), interpreted as a statistical decomposition, estimated an indirect association consistent with sleep quality. Results: Rotating-shift nurses reported the poorest sleep (PSQI = 10.2 ± 2.6; p = 0.003). Physical well-being did not differ significantly across shift types (p = 0.08), although rotating-shift nurses had the lowest mean physical scores (24.3 ± 4.4). The rotat-ing-shift subgroup was small (n = 16), limiting precision. The mediation analysis was statistically consistent with an indirect association between rotating shifts and physical well-being via sleep quality (ACME = −1.85, 95% CI −3.05 to −0.88; p < 0.001), while the proportion of the total association was imprecisely estimated. Conclusions: In this single-site cross-sectional sample, rotating night shifts were associated with poorer sleep and, on average, lower physical well-being; patterns were statistically consistent with an indirect association via sleep quality. Because exposure, mediator, and outcome were measured concurrently, these findings are hypothesis-generating and do not estab-lish causality
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