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The electrode was adjusted until MSNA was recorded after insertion. Identification of MSNA was based on the following criteria: spontaneous burst discharge synchronized with the heartbeat and enhanced by Valsalva maneuver or breath holding, but showing no change in response to sensory stimuli, such as a loud noise or cutaneous touch (Delius et?al. 1972; Vallbo et?al. 1979; Fagius and Wallin 1980). Furthermore, we asked the subjects to hold their breath to identify MSNA at the middle phase during the exercise-spontaneous breathing and after exercise (at least 5?sec). The neurogram was fed to a differential amplifier and amplified 100,000 times through a band-pass filter (700�C2000?Hz). The neurogram was full-wave rectified and integrated JNJ26481585 by a capacitance-integrated circuit with a time constant of 0.1?sec. The mean voltage neurogram was continuously digitized through an analog-to-digital converter with a sampling frequency of 200?Hz for storage on a computer in a manner similar to that for used respiratory and cardiovascular variables. MSNA bursts were identified from the mean voltage neurogram using a customized computer program-assisted inspection (Katayama et?al. 2011, 2012, 2013, 2014), which learn more accounted for the latency from the ECG-R wave to the sympathetic burst (Fagius and Wallin 1980). MSNA was quantified as burst frequency (BF, bursts/min) and burst incidence (BI, bursts/100HR) (Katayama et?al. 2011, 2012, 2013, 2014). Since electromyographic efferent and afferent nerve activities altered the baseline of the integrated neurogram during dynamic leg cycling in most recordings (Saito et?al. 1993; Katayama et?al. 2011, 2012, 2013, 2014), we could not calculate MSNA burst amplitude and total activity. Statistical analysis Values are expressed as means?��?SE. The respiratory and cardiovascular variables, and MSNA BF values were averaged every 1?min throughout the experiment. The assumption of normal distribution for all data was verified using a Kolmogorov-Smirnov test. Changes in parameters during the experiment in each trial were analyzed using (-)-p-Bromotetramisole Oxalate a Dunnett's test, that is, vs. at 5?min at the rest-spontaneous breathing or vs. at 5?min during the exercise-spontaneous breathing. Comparisons of parameters between the nonresistance and resistance trials were performed using paired t-test (parametric test) when the distribution was regular. When the distribution was not regular, a Wilcoxon test (nonparametric test) was used. The StatView (5.0; SAS Institute, Tokyo, Japan) and the SPSS (11.5; SPSS, Tokyo, Japan) statistical packages were used for the analyses. A P?
