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Frequencies of isolated twitches, maximum tetanic contraction and tetanus during fatigue protocol were 0.2, 200 and 100 Hz respectively.
Based on Chan and Head (2010), isolated twitches (0.2Hz) were generated over a 2-min period, followed by a maximum tetanic contraction (induced for 2-s) in each muscle (at 200Hz) [ 33].
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Muscles were adjusted to the optimum length for the development of maximum isometric tetanic contraction.
Square wave pulses (0.2 ms duration) generated by a personal computer with a Labview board (Model PCI-MIO-16E-4, LabvInc.Inc., Austin, TX, USA) were amplified (Acurus Power Amplifier Model A250, Dobbs Ferry, NY, USA) to increase and sustain current intensity to a sufficient level to produce a maximum isometric tetanic contraction.
Maximum isometric tetanic contractions were produced by square wave pulses of 0.2-ms duration, a voltage slightly greater than that required to produce a maximum twitch (usually ∼70 V), and a frequency of 100 Hz (McArdle et al., 2001).
Maximum isometric tetanic contractions were produced by square wave pulses of 0.2-ms duration, a voltage slightly greater than that required to produce a maximum twitch (usually ∼ 70 V), and a frequency of 100 Hz (Brooks & Faulkner, 1991; McArdle et al., 2001).
Stimulation intensity was individually adjusted for each limb in order to obtain tetanic contraction or maximum tolerated intensity.
Further, nNOS has been shown to regulate basal microvascular tone in humans [ 52] and knockout of muscle specific nNOS in mice caused reductions in maximum tetanic force and increased susceptibility to contraction induced fatigue [ 35].
Optimum length of the muscle-tendon complex (the length of the muscle-tendon complex at which maximum tetanic force was produced) for isometric contractions was first estimated using twitches, then determined using a protocol consisting of two twitches and one tetanic contraction (pulse duration 100 μsec, tetanic stimulation frequency: 100 Hz, train duration: 400 ms [ 61].
The maximum tetanic force generated immediately prior to the initiation of the subsequent lengthening contraction was recorded and normalized.
The data presented in Fig. 1 and Table 1 show that exposure of muscle to oxypurinol induced a significant depression in maximum tetanic force generation by EDL and soleus muscles and a reduction in contraction-induced extracellular superoxide anion.
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