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Exercise is presumed to be a potentially helpful smoking cessation adjunct reputed to attenuate the negative effects of deprivation. The present study examined the effectiveness of moderate within-session exercise to reduce 4 key symptoms of smoking deprivation during 3 72-hr nicotine abstinence blocks in both male and female smokers. Forty-nine (25 male, 24 female) sedentary smokers abstained from smoking for 3 consecutive days on 3 separate occasions. At each session, smokers’ abstinence-induced craving, cue-induced craving, negative mood, and withdrawal symptom severity were assessed prior to and after either exercise (a.m. exercise, p.m. exercise) or a sedentary control activity (magazine reading). Abstinence-induced craving and negative mood differed as a function of condition, F(2, 385) = 21, p < .0001; and, F(2, 385) = 3.38, p = .03. Planned contrasts revealed no difference between a.m. and p.m. exercise, but exercise overall led to greater pre-post reduction in abstinence-induced craving, t(385) = 6.23, p < .0001, effect size Cohen’s d = 0.64; and negative mood, t(385) = 2.25, p = .03, d = 0.23. Overall exercise also led to a larger pre-post reduction in cue-induced craving in response to smoking cues, F(2, 387) = 8.94, p = .0002; and withdrawal severity, F(2, 385) = 3.8, p = .02. Unlike the other 3 measures, p.m. exercise reduced withdrawal severity over control, t(385) = 2.64, p = .009, d = 0.27, whereas a.m. exercise did not. The results support the clinical potential of exercise to assist smokers in managing common and robust negative symptoms experienced during the first 3 days of abstinence.