The load during the test was 7.5% of the volunteer’s body mass. Participants were instructed to remain seated throughout the test. The electromyographic activity of each muscle was examined between the second and eighth seconds of each maximum bout, and the highest peak amplitude found, expressed in root mean square (RMS), was used as the normalization factor. Electromyographic activity was monitored continuously during the tests in both experimental conditions (CAF or PLA) using an eight-channel electromyograph (TeleMyo 2400 T G2 – Noraxon Inc., USA). The sampling frequency for EMG records was 2000 Hz and the factor of common-mode rejection CUDC-907 price ratio was greater than 95 dB. The muscles examined SGC-CBP30 cell line were the

superficial quadriceps femoris (QF), RF, VM and VL. The signal was recorded following the recommendations by ISEK. After site preparation by shaving,

cleansing with alcohol and curettage to reduce skin impedance, active electrodes (TeleMyo 2400 – Noraxon Inc., USA) were fixed to the skin, with inter-electrode distance (center to center) of two centimeters. The reference electrode was Integrin inhibitor positioned over the iliac crest. The location of the anatomical landmarks for electrode placement followed the standardization proposed by SENIAM [19]. Analysis and processing of the EMG signal RMS (μV) values were averaged for each 30-s period and were used for the analysis of electromyographic signals from RF, VM, and VL muscles and the integrated

QF [(RF + VM + VL) / 3]. Data were processed using a mathematical simulation environment (Matlab 7.0 – MathWorks ®, South Natick, MA, USA). To obtain the values expressed in RMS, raw EMG signals were digitally filtered, using a band-pass filter of 20Hz and 500Hz, according to the procedures proposed by Dantas et al. [20]. Measurement of perceived exertion All subjects were instructed to report their perceived exertion according to the 6–20 point Borg scale [21] at each 2 km of exercise. From these data, we determined the intercept on the y axis (y-intercept), the Y-27632 datasheet coefficient of determination (R2) and the slope between the time and the individual perceived exertion values attributed during each test obtained by linear regression analysis. Psychological-motivational changes On test days, subjects responded to the Brunel Mood Scale (BRUMS) when they arrived and after the experimental trial. This questionnaire was used for the detection of mood based on 24 questions, stratified into six areas, namely: confusion, anger, depression, fatigue, tension and vigor. Each domain score was normalized by the score obtained prior to the exercise by subtracting the scores at the end of the trial from the scores before the trial. Heart rate During all testing protocols HR was monitored and recorded in RR intervals (ms) and beats per minute (bpm), using a heart rate monitor (Polar RS800CX – Polar®, Kempele, Finland).