## 2 Materials and Methods  
A total of 76 competitive athletes, consisting of 16 females (mean \(\pm\) SD: age 35.4 \(\pm\) 9.67 years, body mass 58.1 \(\pm\) 5.31 kg, height 1.66 \(\pm\) 0.08 m) and 60 males (mean \(\pm\) SD: age 40.2 \(\pm\) 10.9 years, body mass 77.8 \(\pm\) 7.95 kg, height 1.78 \(\pm\) 0.16 m), who were actively involved in either cycling (n = 47) or triathons (n = 20), volunteered to participate in this study. These athletes were chosen because of their competitive nature and familiarity with high-intensity performance environments and their involvement in cycling for 3-20 years (male--8.82 \(\pm\) 7.46 years; female--7.06 \(\pm\) 5.80 years). To ensure the reliability of the test results, participants were asked to refrain from engaging in any form of strenuous physical activity within the 24 h leading up to the test in order to minimize fatigue. Additionally, participants were instructed to avoid consuming caffeine or alcohol for at least three hours before reporting to the laboratory, as these substances could potentially affect physiological responses during the test. All participants were fully informed about the objectives of the study, the procedures involved, as well as any potential risks associated with the tests. Each participant provided written informed consent prior to participation. The study protocol was approved by the University of Toronto Review Ethics Board in accordance with the Declaration of Helsinki. Participants were screened using the PAR-Q and athlete consent form in person. PAR-Q is a physical activity readiness questionnaire that is used to determine the safety and possible risks for an individual to begin an exercise program ([PERSON], 2014). The questionnaire has been used in several studies that involve maximum efforts ([PERSON], 2014; [PERSON], 2014). Once the written consent was obtained, all protocols and procedures were thoroughly explained to the participants to ensure full understanding and compliance. To ensure adherence to pre-test guidelines, participants were asked to avoid caffeine and alcohol for at least 24 h prior to the test. Compliance was verified through self-reporting during the pre-test briefing, and participants were asked to confirm their adherence before beginning the test. While no biochemical validation was conducted, this self-report approach is commonly used in similar exercise physiology studies and has been shown to effectively minimize the influence of caffeine and alcohol on performance during maximal tests. Participants exercised on a computer-controlled, electromagnetically braked cycle ergometer (Excalibur Sport; Lode, Groningen, The Netherlands) set in isokinetic mode, which maintained a constant cadence regardless of power output. The cadence was fixed at each participant's preferred racing cadence, with a range of 85-120 rpm, based on their individual preferences and racing habits. The participant's preferred cadence was determined during a standardized warm-up phase, where they were instructed to cycle at a self-selected pace for a few minutes. The cadence at which they felt most comfortable and could sustain was recorded and then used as their preferred cadence for the trial. This self-selected cadence was maintained during the baseline phase to ensure consistency across participants. The use of self-selected cadence allows for individual optimization and is commonly employed in cycling and endurance sports protocols [17]. Power output data were sampled by the Lode ergometer at 6 Hz and subsequently exported from the ergometer software for analysis. Prior to each trial, the participant completed their regular race warm-up routine, which lasted between 10 and 20 min at an intensity of 50-100 W. This warm-up period was followed by 5 min of rest to ensure that the participant was in a rested state before beginning the test. The trial itself began with 1 min of light cycling at less than 100 W, during which the participant was asked to gradually increase their effort in the final 5 s of this phase.