27Acetaminophen has been combined with caffeine for therapeutic purpose, but the effect 28 of co-ingestion of acetaminophen and caffeine on exercise performance has not been 29 investigated. The aim of this study was to determine the effect of isolated and combined 30 ingestion of caffeine and acetaminophen on performance during a 4-km cycling time-trial. In a 31 double-blind, crossover design, eleven men, accustomed to cycling recreationally, completed a 32 4-km cycling time-trial one hour after the ingestion of cellulose (PLA), acetaminophen (20 33 mg . kg -1 body mass, ACT), caffeine (5 mg . kg -1 body mass, CAF) or combined acetaminophen 34 and caffeine (20 and 5 mg . kg -1 body mass, respectively, ACTCAF). The perception of pain and 35 rating of perceived exertion were recorded every 1-km, and electromyography and oxygen 36 uptake were continually recorded and averaged each 1-km. Plasma lactate concentration was 37 measured before and immediately after the trial. The time and mean power during the 4-km 38 cycling time-trial was significantly improved (P < 0.05) in CAF (407.9 ± 24.5 s, 241.4 ± 16.1 39 W) compared to PLA (416.1 ± 34.1 s, 234.1 ± 19.2 W) and ACT (416.2 ± 26.6 s, 235.8 ± 19.7 40 W). However, there was no difference between ACTCAF (411.6 ± 27.7 s, 238.7 ± 18.7 W) and 41 the other conditions (P > 0.05). The perception of pain, rating of perceived exertion, 42 electromyography, oxygen uptake, and plasma lactate were similar across the conditions (P > 43 0.05). In conclusion, caffeine but not acetaminophen increases power output ultimately 44 increasing performance during a 4-km cycling time-trial. 45 46 47 48 49 50 51 52 53 3 54 Introduction 55 56During a self-paced, high-intensity cycling time-trial (e.g., 4-km cycling TT), the 57 exercise intensity must be strictly regulated to avoid an exacerbated early accumulation of 58 metabolites that can lead to fatigue [1][2][3]. A disturbance in the intramuscular metabolic milieu 59 (i.e., accumulation of the H + , ADP, AMP and Pi) at the beginning of a high-intensity cycling 60 TT, provoked by increased muscle recruitment (as inferred from electromyography signals, 61 EMG) ultimately increasing power output (PO) [2,3], will activate peripheral sensory nerve 62 terminals [4,5]. These increased afferent signals might result in increased perception of pain 63 [6], leading to a reduction in PO [2,3]. 64 65Acetaminophen (commonly known as paracetamol) has recently been introduced as a 66 potential pharmacological agent to increase exercise performance due to its analgesic 67 proprieties [7-9]. The mechanism by which acetaminophen reliefs pain feelings in humans is 68 not fully known, but it has been attributed to the inhibition of the cyclooxygenase enzymes [10-69 13], potentiation of descending serotoninergic pathways [14,15], and modulation of opioid and 70 cannabinoid CB 1 receptors [11,16]. Although there is not a consensus [17,18], several studies 71 have reported improved performance during high-intensity exercises after a single clinical dose
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