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The concentration that produced 50% of maximal effect, the area under the curve, and the maximal effect of U46619, normalized with respect to a reference contraction in response to potassium chloride, were compared.
First, each segment was exposed to 124 mmol/L K+ Krebs solution to elicit a reference contraction.
At the outset of each experiment, K+-rich (80 mM) solution, obtained by substituting an equimolar amount of KCl for NaCl from Krebs-HEPES solution, was applied in order to obtain a reference contraction used to normalize subsequent contractile responses.
sEMG calibration by MVC or by sub-maximal reference contraction is however the preferred calibration procedure [ 26].
Using diaphragm length signal as reference, contraction times estimated with the other three signals were compared with the contraction time estimated with diaphragm length signal.
There was no change in the average EMG amplitude during the reference contraction (i.e., arms 90 degree abducted) from before to after the daily training session, showing that the EMG measurements were stable throughout the day.
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The mean (SD; min-max) reference contractions' signal-to-noise ratios were 48 (6; 38 61) dB for AN, 44 (5; 32 54) dB for UPN, and 31 (3; 22–36) dB for LPN.
Similarly, we found no group differences in absolute rmsEMG amplitude (μV) values during the reference contractions for the muscles (p-values > 0.11).
The EMG recordings were root-mean-square (RMS) converted in 0.1-s periods, adjusted for noise, normalized to submaximal reference contractions, and expressed in % RVE (reference voluntary electrical activity) (Mathiassen et al. 1995).
An important procedural difference between the two studies is that the previous study reported sEMG responses calibrated in absolute units (μV), due to the difficulty of achieving reliable reference contractions of forehead muscles.
EMG epochs covering the entire movement as defined by the kinematic start and stop procedures ± 200 ms were subsequently normalized to the median rmsEMG accomplished at reference contractions (see Additional files 1 and 2 for descriptions) and further analyzed.
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