Vastus lateralis single motor unit EMG at the same absolute torque production at different knee angles.

Single motor unit electromyographic (EMG) activity of the knee extensors was investigated at different knee angles with subjects (n = 10) exerting the same absolute submaximal isometric torque at each angle. Measurements were made over a 20 degrees range around the optimum angle for torque production (AngleTmax) and, where feasible, over a wider range (50 degrees ). Forty-six vastus lateralis (VL) motor units were recorded at 20.7 +/- 17.9 %maximum voluntary contraction (%MVC) together with the rectified surface EMG (rsEMG) of the superficial VL muscle. Due to the lower maximal torque capacity at positions more flexed and extended than AngleTmax, single motor unit recruitment thresholds were expected to decrease and discharge rates were expected to increase at angles above and below AngleTmax. Unexpectedly, the recruitment threshold was higher (P < 0.05) at knee angles 10 degrees more extended (43.7 +/- 22.2 N.m) and not different (P > 0.05) at knee angles 10 degrees more flexed (35.2 +/- 17.9 N.m) compared with recruitment threshold at AngleTmax (41.8 +/- 21.4 N.m). Also, unexpectedly the discharge rates were similar (P > 0.05) at the three angles: 11.6 +/- 2.2, 11.6 +/- 2.1, and 12.3 +/- 2.1 Hz. Similar angle independent discharge rates were also found for 12 units (n = 5; 7.4 +/- 5.4 %MVC) studied over the wider (50 degrees ) range, while recruitment threshold only decreased at more flexed angles. In conclusion, the similar recruitment threshold and discharge behavior of VL motor units during submaximal isometric torque production suggests that net motor unit activation did not change very much along the ascending limb of the knee-angle torque relationship. Several factors such as length-dependent twitch potentiation, which may contribute to this unexpected aspect of motor control, are discussed.

Vastus lateralis surface and single motor unit electromyography during shortening, lengthening and isometric contractions corrected for mode-dependent differences in force-generating capacity.

AIM: Knee extensor neuromuscular activity, rectified surface electromyography (rsEMG) and single motor unit EMG was investigated during isometric (60 degrees knee angle), shortening and lengthening contractions (50-70 degrees, 10 degrees s(-1)) corrected for force-velocity-related differences in force-generating capacity. However, during dynamic contractions additional factors such as shortening-induced force losses and lengthening-induced force gains may also affect force capacity and thereby neuromuscular activity. Therefore, even after correction for force-velocity-related differences in force capacity we expected neuromuscular activity to be higher and lower during shortening and lengthening, respectively, compared to isometric contractions. METHODS: rsEMG of the three superficial muscle heads was obtained in a first session [10 and 50% maximal voluntary contraction (MVC)] and additionally EMG of (46) vastus lateralis motor units was recorded during a second session (4-76% MVC). Using superimposed electrical stimulation, force-generating capacity for shortening and lengthening contractions was found to be 0.96 and 1.16 times isometric (Iso) force capacity respectively. Therefore, neuromuscular activity during submaximal shortening and lengthening was compared with isometric contractions of respectively 1.04Iso (=1/0.96) and 0.86Iso (=1/1.16). rsEMG and discharge rates were normalized to isometric values. RESULTS: rsEMG behaviour was similar (P > 0.05) during both sessions. Shortening rsEMG (1.30 +/- 0.11) and discharge rate (1.22 +/- 0.13) were higher (P < 0.05) than 1.04Iso values (1.05 +/- 0.05 and 1.03 +/- 0.04 respectively), but lengthening rsEMG (1.05 +/- 0.12) and discharge rate (0.90 +/- 0.08) were not lower (P > 0.05) than 0.86Iso values (0.76 +/- 0.04 and 0.91 +/- 0.07 respectively). CONCLUSION: When force-velocity-related differences in force capacity were taken into account, neuromuscular activity was not lower during lengthening but was still higher during shortening compared with isometric contractions.

Changes in force, surface and motor unit EMG during post-exercise development of low frequency fatigue in vastus lateralis muscle.

We investigated the effects of low frequency fatigue (LFF) on post-exercise changes in rectified surface EMG (rsEMG) and single motor unit EMG (smuEMG) in vastus lateralis muscle (n = 9). On two experimental days the knee extensors were fatigued with a 60-s-isometric contraction (exercise) at 50% maximal force capacity (MFC). On the first day post-exercise (15 s, 3, 9, 15, 21 and 27 min) rsEMG and electrically-induced (surface stimulation) forces were investigated. SmuEMG was obtained on day two. During short ramp and hold (5 s) contractions at 50% MFC, motor unit discharges of the same units were followed over time. Post-exercise MFC and tetanic force (100 Hz stimulation) recovered to about 90% of the pre-exercise values, but recovery with 20 Hz stimulation was less complete: the 20-100 Hz force ratio (mean +/- SD) decreased from 0.65+/-0.06 (pre-exercise) to 0.56+/-0.04 at 27 min post-exercise (P<0.05), indicative of LFF. At 50% MFC, pre-exercise rsEMG (% pre-exercise maximum) and motor unit discharge rate were 51.1 +/- 12.7% and 14.1 +/- 3.7 (pulses per second; pps) respectively, 15 s post-exercise the respective values were 61.4 +/- 15.4% (P<0.05) and 13.2 +/- 5.6 pps (P>0.05). Thereafter, rsEMG (at 50% MFC) remained stable but motor unit discharge rate significantly increased to 17.7 +/- 3.9 pps 27 min post-exercise. The recruitment threshold decreased (P<0.05) from 27.7 +/- 6.6% MFC before exercise to 25.2 +/- 6.7% 27 min post-exercise. The increase in discharge rate was significantly greater than could be expected from the decrease in recruitment threshold. Thus, post-exercise LFF was compensated by increased motor unit discharge rates which could only partly be accounted for by the small decrease in motor unit recruitment threshold.

Voluntary drive-dependent changes in vastus lateralis motor unit firing rates during a sustained isometric contraction at 50% of maximum knee extension force.

The purpose of the present study was to relate the expected inter-subject variability in voluntary drive of the knee extensor muscles during a sustained isometric contraction to the changes in firing rates of single motor units. Voluntary activation, as established with superimposed electrical stimulation was high (range: 91-99%, n=8) during a short maximal contraction, but was lower (range: 69-100%) in most subjects at the point of force failure during a sustained (49.1+/-10.1 s) fatiguing contraction at 50% of maximum force. On a different experimental day the firing behaviour of 27 single motor units was recorded with wire electrodes in the vastus lateralis muscle, 24 of which could be monitored from the time of recruitment to the point of force failure (53.6+/-9.8 s). Motor unit firing behaviour differed considerably among subjects. During the second half of the sustained, fatiguing contraction the changes in firing rate firing rate variability of early recruited units ranged from -10% to +100% and from -50% to +160% respectively among subjects. There were significant positive linear relations between voluntary activation, on the one hand, and rectified surface electromyogram (rsEMG, r=0.82), the changes in motor unit firing rate ( r=0.49) and firing rate variability ( r=0.50) towards the point of force failure on the other. The present data suggest that differences in voluntary drive that appear among subjects during fatigue may be an important determinant of motor unit firing behaviour.

Efficient identification of novel HLA-A(*)0201-presented cytotoxic T lymphocyte epitopes in the widely expressed tumor antigen PRAME by proteasome-mediated digestion analysis.

We report the efficient identification of four human histocompatibility leukocyte antigen (HLA)-A(*)0201-presented cytotoxic T lymphocyte (CTL) epitopes in the tumor-associated antigen PRAME using an improved "reverse immunology" strategy. Next to motif-based HLA-A(*)0201 binding prediction and actual binding and stability assays, analysis of in vitro proteasome-mediated digestions of polypeptides encompassing candidate epitopes was incorporated in the epitope prediction procedure. Proteasome cleavage pattern analysis, in particular determination of correct COOH-terminal cleavage of the putative epitope, allows a far more accurate and selective prediction of CTL epitopes. Only 4 of 19 high affinity HLA-A(*)0201 binding peptides (21%) were found to be efficiently generated by the proteasome in vitro. This approach avoids laborious CTL response inductions against high affinity binding peptides that are not processed and limits the number of peptides to be assayed for binding. CTL clones induced against the four identified epitopes (VLDGLDVLL, PRA(100-108); SLYSFPEPEA, PRA(142-151); ALYVDSLFFL, PRA(300-309); and SLLQHLIGL, PRA(425-433)) lysed melanoma, renal cell carcinoma, lung carcinoma, and mammary carcinoma cell lines expressing PRAME and HLA-A(*)0201. This indicates that these epitopes are expressed on cancer cells of diverse histologic origin, making them attractive targets for immunotherapy of cancer.

Effects of high-frequency initial pulses and posttetanic potentiation on power output of skeletal muscle.

The effects of high-frequency initial pulses (HFIP) and posttetanic potentiation on mechanical power output during concentric contractions were examined in the in situ medial gastrocnemius of the rat with an intact origin on the femur and blood supply. Stimulation of the muscle was performed via the severed sciatic nerve. In the experiments, HFIP or the potentiating tetanus was followed by a stimulation of 80, 120, or 200 Hz. The results showed that both HFIP and the tetanus increased power output at high contraction velocities (>75 mm/s) when followed by a train of 80 or 120 Hz (200 Hz resulted in no effects). Mechanical power output was increased maximally by HFIP to 120 and 168% by the tetanus. Furthermore, when HFIP or the tetanus were followed by a train of 80 Hz, the peak power in the power-velocity curve tended to be shifted to a higher velocity.

Stimulation frequency-dependent reductions in skeletal muscle force and speed in creatine kinase-deficient mice.

Force and speed parameters were obtained from isometric contractions at different stimulation frequencies of creatine kinase-deficient and wildtype in situ mouse medial gastrocnemius muscles. The absence of creatine kinase did not affect force production at higher stimulation frequencies. However, at frequencies below 140 Hz, forces were lower than the controls (P < 0.05); at the lowest frequency applied (80 Hz) the force was reduced to approximately 60% compared with the wildtype muscles. In contrast, twitch force was not affected. When the contractions were preceded by a brief tetanus (50 ms), the effects of lacking creatine kinase on force production were more pronounced; at 80 Hz stimulation isometric force was further reduced to 66.5 +/- 6.2% (mean +/- SD; n=5) of the single contractions of the deficient muscles and to approximately 42% of the wildtype muscles. Twitch force was now also reduced (by approximately 50%) after the tetanus. The speed of the muscles was not affected in the single contractions. However, after a preceding tetanus, the rate of force rise was reduced by approximately 14% at high frequencies of stimulation. With decreasing frequencies (below 250 Hz), the reduction in speed became more pronounced; at 80 Hz the rate in the creatine kinase-deficient muscles was only 55.2 +/- 3.9% (mean +/- SD; n=5) of the wildtype muscles. No effects of the deficiency were found for the half relaxation times. The data suggest that an impaired creatine kinase system leads to lower activation levels at submaximal stimulation frequencies, possibly by a reduction in Ca2+-release during repetitive stimulation. Similar effects may be expected in normal fatigued muscle when phosphocreatine is depleted.

Muscle function in a patient with Brody's disease.

Adductor pollicis muscle function of a 21-year-old man with genetically confirmed Brody's disease (sarcoplasmic reticulum [SR] -Ca2+ATPase deficiency) was investigated to study the possible effects of reduced SR-Ca2+ATPase activity on muscle relaxation and force production. Following maximal electrical activation of the ulnar nerve, tetanic muscle half-relaxation time was greater in the patient (246 +/- 10 ms) than control subjects (97 +/- 4 ms, n = 8). During repetitive activation, there was a similar decline in maximal shortening velocity in the patient and controls, indicating a comparable reduction in cross-bridge cycling rate. The finding that the slowing of relaxation was greater in the patient (329 ms versus 138 +/- 20 ms) suggests that there was a further reduction of SR-Ca2+ATPase activity in the patient's muscle during fatigue. Following a voluntary contraction, involuntary activity of the antagonist muscles facilitated force decline and masked the impaired relaxation in the patient. This antagonist-induced relaxation indicates that it might be difficult to establish impaired muscle relaxation with voluntary contractions.