Determination of fatigue in the electrically stimulated quadriceps muscle and relative effect of ischaemia

The creation of muscle fatigue using surface electrical stimulation represents a highly reproducible phenomenon in spinal cord injured patients. The torque output was recorded as a function of time. The fatigue curves recorded over 110s exhibited three main parts: first, a plateau of short duration, followed by a more or less steep slope and then a second plateau which was maintained for a long time. This phenomenon was fitted using an exponential equation which had been developed and four parameters introduced that outlined to the muscle behaviour. A set of fatigue indices was defined to characterize the asymptotic value, the slope, the coordinates of the inflexion point, the time constant and the vertical amplitude of the curve recorded. Two populations were studied; a group of 11 thoracic level of injury paraplegic patients and a group of 10 able-bodied control subjects. The computed coefficients of determination, r², were of very high values (0.99). Therefore, fatigue indices gave reliable information. Torque output did not differ between the two populations until 25 s had elapsed, but from 30s onwards it was markedly lower in paraplegics. The residual torque output was 21.1 ± 10.6% in the paraplegic group while it was 58.5 ± 8.9% in the control group. The effect of blood supply in the production of fatigue was also studied by repeating the same test using a tourniquet at the groin level. The residual torque became 14.7 ± 2.3% in the paraplegic group and 42.9 ± 6.3% in the control group. This test permitted the determination at a gross level of the different metabolic phases and therefore the recruitment of the different populations of muscle fibres within the quadriceps. Fast fatiguable fibres (type FF) produced the maximum output during the first phase, then fast resistant (type FR) explained the slope, and slow fibres (type S) were responsible for the second plateau. The effects of the tourniquet were obvious on the fatigue indices in both populations. In the spinal cord injured patients, there appeared to be a lack of slow fibres, probably due to disuse. Fatigue appeared to be independent from blood flow in paraplegics.     

Near infrared monitoring of human skeletal muscle oxygenation during forearm ischemia

Changes in tissue oxygenation of forearm muscles were measured by near infrared (NIR) spectrophotometry in 10 healthy adults during tourniquet ischemia and venous outflow restriction. Muscle O2 stores were depleted rapidly by forearm ischemia manifest by a progressive decrease in tissue oxyhemoglobin and oxymyoglobin over 4-5 min. Muscle ischemia significantly decreased the oxidation level of cytochrome aa3, to below resting base line after only 1.5 min, and the enzyme became fully reduced after 6.5 min. After 8 min of ischemia, tourniquet release was accompanied by a transient increase in muscle blood volume due to influx of oxyhemoglobin. The cytochrome aa3 oxidation level increased above resting base line within 1 min after tourniquet release. Transcutaneous PO2 measurements recorded simultaneously from the same forearm correlated poorly with the kinetics of O2 availability and cytochrome oxidation in the underlying muscle tissue; this was not unexpected because overlying skin did not contribute significantly to NIR muscle signals. Venous outflow restriction without inflow obstruction increased muscle deoxyhemoglobin and tissue blood volume but did not change muscle O2 stores or cytochrome aa3 oxidation level. The ability of the NIR technique to detect dynamic trends in tissue oxygenation reveals that muscle O2 is rapidly consumed during tourniquet ischemia and rapidly restored by hyperemic responses after brief ischemia.     

Mechanical properties of the latissimus dorsi muscle after cyclic training.

Cardiomyoplasty is a procedure developed to improve heart performance in patients suffering from congestive heart failure. The latissimus dorsi (LD) muscle is surgically wrapped around the failing ventricles and stimulated to contract in synchrony with the heart. The LD muscle is easily fatigued and as a result is unsuitable for cardiomyoplasty. For useful operation as a cardiac-assist device, the fatigue resistance of the LD muscle must be improved while retaining a high power output. The LD muscle of rabbits was subjected to a training regime in which cyclic work was performed. Training transformed the fiber-type composition from approximately equal proportions of fast oxidative glycolytic (FOG) and fast glycolytic (FG) fibers to one composed of almost entirely of FOG with no FG, which increased fatigue resistance while retaining rapid contraction kinetics. Muscle mass and cross-sectional area increased but power output decreased, relative to control muscles. This training regime represents a significant improvement in terms of preserving muscle mass and power compared with other training regimes, while enhancing fatigue resistance, although some fiber damage occurred. The power output of the trained LD muscle was calculated to be sufficient to deliver a significant level of assistance to a failing heart during cardiomyoplasty.     

Diaphragm muscle fatigue resistance during postnatal development.

Changes in the contractile and fatigue properties of the cat diaphragm muscle were examined during the first 6 wk of postnatal development. Both twitch contraction time and half-relaxation time decreased progressively with age. Correspondingly, the force-frequency curve was shifted to the left early in development compared with adults. The ratio of peak twitch force to maximum tetanic force decreased with age. Fatigue resistance of the diaphragm was highest at birth and then progressively decreased with age. At birth, most diaphragm muscle fibers stained darkly for myofibrillar adenosinetriphosphatase after alkaline preincubation and thus would be classified histochemically as type II. During subsequent postnatal development, the proportion of type I fibers (lightly stained for adenosinetriphosphatase) increased while the number of type II fibers declined. At birth, type I fibers were larger than type II fibers. The size of both fiber types increased with age, but the increase in cross-sectional area was greater for type II fibers. On the basis of fiber type proportions and mean cross-sectional areas, type I fibers contributed 15% of total muscle mass at birth and 25% in adults. Thus postnatal changes in diaphragm contractile and fatigue properties cannot be attributed to changes in the relative contribution of histochemically classified type I and II fibers. However, the possibility that these developmental changes in diaphragm contractile and fatigue properties correlated with the varying contractile protein composition of muscle fibers was discussed.     

Capillary and fiber size interrelationships in regenerating rat soleus muscle after ischemia: a quantitative study.

The present study examines the influence of ischemia on the muscle fibers and capillarization in rats. Muscle ischemia was achieved by a pneumatic tourniquet at a pressure of 300 mm Hg for 2, 4 and 6 h (groups I, II and III, respectively) to the right hindlimb above the knee. Numerous regenerative fibers were seen at 4 and, especially, 8 and 12 days after ischemia in groups II and III. The quantitative data revealed a significant decrease in the size of muscle fibers (regenerative fibers) in ischemic skeletal muscle, with a concomitant increase in fiber density. The capillary to fiber ratio shows a decrease at 4, 8 and 12 days after ischemia in the three experimental groups: in group I because of a decrease in capillary density; in groups II and III because of an increase in fiber density with respect to capillary density.     

Ischemia reperfusion injury of the skeletal muscle after selective deafferentation

The present study analyzes the effect of selective deafferentation on the reperfusion injury of the skeletal muscle when nociceptive sensory fibers of the left sciatic nerve are selectively damaged by capsaicin pretreatment in a rat model following tourniquet ischemia (ISC) applied for 30 min, 1 h, and 2 h on the left hind limb. The isometric tetanic contractile force of the extensor digitorum longus (EDL) muscle was measured after 1 h, and 1, 3, or 7 days of reperfusion. Contractile force of the damaged muscle was compared to the intact contralateral muscle. In another group, ISC was used without capsaicin pre-treatment. After 30 min of ISC, there was no difference between deafferented and non-pretreated groups. Following 1 h ISC, with the exception of 1 h reperfusion, the non-pretreated group produced stronger contractions than the deafferented group. After 2 h ISC, the contractile force of the deafferented muscle was significantly stronger compared to the non-deafferented muscle force at all reperfusion times. In conclusions, it was found that the absence of peptidergic sensory fibers after long-lasting (2 h) ischemia is beneficial in reperfusion injury, whereas the absence of vasodilator peptides has unfavorable effects if tissue damage is milder (after 1 h ischemia).     

Assessment of muscle fatigue in low level monotonous task performance during car driving.

Knowledge of the muscle activation and the development of muscle fatigue may provide more inside in the effects of long-term driving in the occurrence of health problems in the neck/shoulder/back area. The basic assumption behind fatigue detection with electromyography (EMG) is an increase in the EMG amplitude and a decrease of the mean frequency (MF). This study aimed at checking this assumption in monotonous task performance with low level activity during car driving. Surface electromyography was captured from left and right trapezius and deltoid muscles, during a repetitive, non-continuous, driving task (gearing and steering) and the active parts were separated from the non-active parts. Muscle stiffness was reported by more than half of the subjects after a 1 h drive. Only for the active parts a significant decrease of the MF was seen. But also the EMG amplitude decreased significantly. Two possible mechanisms are posted in literature for this finding: no extra recruitment of motor units (MU) and potentiation of muscle fibers. Literature also hypothesizes that low-force occupational work engages only a fraction of the MU available for recruitment and that these units are selectively type I muscle fibers (Cinderella fibers). Initiators of this phenomenon are probably the time lag between activations and the stress from driving and vibration exposure.     

The interactive effects of fatigue and pH on the ionic conductance of frog sartorius muscle fibers

The effects of fatigue on the membrane conductance of frog sartorius muscle at the resting potential and during an action potential were studied. When muscles were exposed to an extracellular pH of 8.0 the membrane conductance at the resting potential increased during fatigue by about 20% and returned to prefatigue level in about 20 min. The membrane conductance of muscle fibers exposed to pH 6.4 was about three times less than that of pH 8.0 and decreased further during fatigue. Furthermore, the recovery of a normal membrane conductance was slow at pH 6.4. Both the inward, depolarizing and the outward, repolarizing currents during the action potential are reduced in fatigue. In each case the effect is greater at pH 6.4 than at 8.0 and recovery towards normal values is slower at pH 6.4. It is concluded that the ionic conductance of the sarcolemmal membrane at the resting potential and during an action potential are modified by fatigue and that these changes are modulated by pHo.     

Effects of fatigue on sarcoplasmic reticulum and myofibrillar properties of rat single muscle fibers.

Force decline during fatigue in skeletal muscle is attributed mainly to progressive alterations of the intracellular milieu. Metabolite changes and the decline in free myoplasmic calcium influence the activation and contractile processes. This study was aimed at evaluating whether fatigue also causes persistent modifications of key myofibrillar and sarcoplasmic reticulum (SR) proteins that contribute to tension reduction. The presence of such modifications was investigated in chemically skinned fibers, a procedure that replaces the fatigued cytoplasm from the muscle fiber with a normal medium. Myofibrillar Ca(2+) sensitivity was reduced in slow-twitch muscle (for example, the pCa value corresponding to 50% of maximum tension was 6.23 +/- 0.03 vs. 5.99 + 0.05, P < 0.01, in rested and fatigued fibers) and not modified in fast-twitch muscle. Phosphorylation of the regulatory myosin light chain isoform increased in fast-twitch muscle. The rate of SR Ca(2+) uptake was increased in slow-twitch muscle fibers (14.2 +/- 1.0 vs. 19.6 +/- 2. 5 nmol. min(-1). mg fiber protein(-1), P < 0.05) and not altered in fast-twitch fibers. No persistent modifications of SR Ca(2+) release properties were found. These results indicate that persistent modifications of myofibrillar and SR properties contribute to fatigue-induced muscle force decline only in slow fibers. These alterations may be either enhanced or counteracted, in vivo, by the metabolic changes that normally occur during fatigue development.     

Generation, in activation and level of blood kinins during tourniquet shock in rabbits

Traumatic shock was induced by the tourniquet method compressing one thigh during 10 hours. Venous blood samples were taken from control animals, as well as twice in the nervous phase of shock - after application and before removal of the tourniquet, and in the humoral-toxic phase - 1, 3 and 5 hours after tourniquet removal, in groups of 10 animals. Determinations included blood kinin level, and plasma kininogen level, and the activity of kallikreins and kininases in the plasma. It was found that during tourniquet shock a significant change occurred in the whole blood kinin system. Proportionally to the severity of shock the level of free kinins and kallikrein activity increased 3-4, times and the level of kininogen and the activity of kininases decreased, especially 3 hours after tourniquet removal.     

Denervation syndrome in the rat digastric muscle

Excitability parameters of m. digastricus muscle fibers were investigated in anesthetized (40 mg/kg of nembutal) rats 3-5 days after denervation. The number of fibers with high polarization level was increased as was the number of fibers with low and medium level in both bellies of m. digastricus. The differences in the level of polarization recorded in the muscle fibers of the abdominoposterior m. digastricus disappeared after denervation. It is suggested that denervation syndrome in m. digastricus deprived of spindle receptors was similar to that observed in other skeletal muscles.     

Inhibition of shortening velocity of skinned skeletal muscle fibers in conditions that mimic fatigue

The mechanisms responsible for the inhibition of shortening velocity that occurs during muscle fatigue have not been completely elucidated. Phosphorylation of the myosin regulatory light chain (RLC) occurs during heavy use; however, previous reports on its role in affecting velocity have been equivocal. To further understand the process of fatigue, we varied the levels of myosin RLC phosphorylation (from 10 to >50%) and the concentrations of protons (from pH 7 to 6.2) and phosphate (from 5 to 30 mM), all of which change during fatigue. We measured the mechanics of permeable rabbit psoas fibers at a temperature closer to physiological (30 degrees C), using a temperature jump protocol to briefly activate the fibers at the higher temperature to preserve sarcomere homogeneity. Although lowered pH alone had an effect on velocity, it was the three factors together, i.e., high phosphorylation, low pH, and high phosphate, that acted synergistically to inhibit fiber velocity by approximately 40%. Our data demonstrate that in conditions that simulate physiological muscle fatigue, myosin phosphorylation does contribute to the inhibition of contraction velocity of fully activated fast muscle fibers.     

Effect of varied extracellular PO2 on muscle performance in Xenopus single skeletal muscle fibers.

The purpose of this study was to examine the development of fatigue in isolated, single skeletal muscle fibers when O2 availability was reduced but not to levels considered rate limiting to mitochondrial respiration. Tetanic force was measured in single living muscle fibers (n = 6) from Xenopus laevis while being stimulated at increasing contraction rates (0.25, 0.33, 0.5, and 1 Hz) in a sequential manner, with each stimulation frequency lasting 2 min. Muscle fatigue (determined as 75% of initial maximum force) was measured during three separate work bouts (with 45 min of rest between) as the perfusate PO2 was switched between values of 30 +/- 1.9, 76 +/- 3.0, or 159 Torr in a blocked-order design. No significant differences were found in the initial peak tensions between the high-, intermediate-, and low-PO2 treatments (323 +/- 22, 298 +/- 27, and 331 +/- 24 kPa, respectively). The time to fatigue was reached significantly sooner (P < 0.05) during the 30-Torr treatment (233 +/- 39 s) compared with the 76- (385 +/- 62 s) or 159-Torr (416 +/- 65 s) treatments. The calculated critical extracellular PO2 necessary to develop an anoxic core within these fibers was 13 +/- 1 Torr, indicating that the extracellular PO2 of 30 Torr should not have been rate limiting to mitochondrial respiration. The magnitude of an unstirred layer (243 +/- 64 micron) or an intracellular O2 diffusion coefficient (0.45 +/- 0.04 x 10(-5) cm2/s) necessary to develop an anoxic core under the conditions of the study was unlikely. The earlier initiation of fatigue during the lowest extracellular PO2 condition, at physiologically high intracellular PO2 levels, suggests that muscle performance may be O2 dependent even when mitochondrial respiration is not necessarily compromised.     

Depressed contractile performance and reduced fatigue resistance in single skinned fibers of soleus muscle after long-term disuse in rats

Long-term disuse results in atrophy in skeletal muscle, which is characterized by reduced functional capability, impaired locomotor condition, and reduced resistance to fatigue. Here we show how long-term disuse affects contractility and fatigue resistance in single fibers of soleus muscle taken from the hindlimb immobilization model of the rat. We found that long-term disuse results in depression of caffeine-induced transient contractions in saponin-treated single fibers. However, when normalized to maximal Ca(2+)-activated force, the magnitude of the transient contractions became similar to that in control fibers. Control experiments indicated that the active force depression in disused muscle is not coupled with isoform switching of myosin heavy chain or troponin, or with disruptions of sarcomere structure or excessive internal sarcomere shortening during contraction. In contrast, our electronmicroscopic observation supported our earlier observation that interfilament lattice spacing is expanded after disuse. Then, to investigate the molecular mechanism of the reduced fatigue resistance in disused muscle, we compared the inhibitory effects of inorganic phosphate (Pi) on maximal Ca(2+)-activated force in control vs. disused fibers. The effect of Pi was more pronounced in disused fibers, and it approached that observed in control fibers after osmotic compression. These results suggest that contractile depression in disuse results from the lowering of myofibrillar force-generating capacity, rather than from defective Ca(2+) mobilization, and the reduced resistance to fatigue is from an enhanced inhibitory effect of Pi coupled with a decrease in the number of attached cross bridges, presumably due to lattice spacing expansion.     

Effect of corticosteroids on diaphragm fatigue, SDH activity, and muscle fiber size.

The influence of dexamethasone on diaphragm (DIA) fatigue, oxidative capacity, and fiber cross-sectional areas (CSA) was determined in growing hamsters. One group received dexamethasone by daily subcutaneous injection for 21 days (D animals), while pair-weight (P) and free-eating controls (CTL) received saline subcutaneously. Isometric contractile properties of the DIA were determined in vitro by supramaximal direct muscle stimulation in the presence of curare. DIA fatigue resistance was determined through repetitive stimulation at 40 pulses/s for 2 min. A computer-based image-processing system was used to histochemically determine muscle fiber-type proportions, CSA, and succinate dehydrogenase activities. The medial gastrocnemius muscle (MG) was used as a limb muscle control, with histochemical studies being performed on both the superficial (s) and deep/red (r) portions. Dexamethasone markedly attenuated the normal increment in body weight over the 3-wk period. DIA fatigue resistance was significantly reduced in the D compared with CTL and P animals. Dexamethasone had no effect on fiber-type proportions of the DIA or MGr (MGs contained only type II fibers). In the DIA, the CSA of type II fibers was reduced 33% in D and 18.5% in P animals compared with CTL. Although no significant atrophy was noted in the type I DIA fibers of either D or P animals, a trend toward significance was noted in D animals compared with CTL. In the MGs, the CSA of type II fibers was reduced 33% in D and 16.5% in P animals compared with CTL. Significant atrophy of type I and II fibers of the MGr was noted in D animals compared with CTL (33.8 and 35% reductions, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship between numbers and frequencies of stimuli in human muscle fatigue

The effect of stimulus frequency on the rate of muscle fatigue has been studied on dorsiflexor muscles of the human ankle. It was found that significantly fewer stimuli were required to abolish twitch and tetanic torque when the stimuli were delivered at 15 Hz rather than 30 Hz. At both stimulus frequencies twitch torque disappeared before tetanic torque. The difference in numbers of stimuli required for fatigue was not due to impaired excitation of muscle fibers at either of the two frequencies. At both stimulating frequencies, twitch fatigue appeared to be due to a defect in excitation-contraction coupling and/or the contractile machinery.     

Microelectrophysiological analysis of the fiber components of the rat digastric muscle

Membrane potential at rest (MP), action potential (AP), critical level of depolarization (CLD) and latent period (LP) of different muscle fibers were studied in two bellies of digastric muscle. Even and chaotic distribution of different muscle fibers was observed in the anterior and posterior belly, respectively. It is believed that electrophysiological data correspond to the results of histological analysis of muscle fibers in digastric muscle.     

Intracellular pH during sequential, fatiguing contractile periods in isolated single Xenopus skeletal muscle fibers.

The purpose of the present study was to test the hypothesis that a preceding contractile period in isolated single skeletal muscle fibers would attenuate the decrease in pH during an identical, subsequent contractile period, thereby reducing the rate of fatigue. Intact single skeletal muscle fibers (n = 9) were isolated from Xenopus lumbrical muscle and incubated with the fluorescent cytosolic H+ indicator 2',7'-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) AM for 30 min. Two identical contractile periods were performed in each fiber, separated by a 1-h recovery period. Force and intracellular pH (pHi) fluorescence were measured simultaneously while fibers were stimulated (tetanic contractions of 350-ms trains with 70-Hz stimuli at 9 V) at progressively increasing frequencies (0.25, 0.33, 0.5, and 1 contraction/s) until the development of fatigue (to 60% initial force). No significant difference (P < 0.05) was observed between the first and second contractile periods in initial force development, resting pHi, or time to fatigue (5.3 +/- 0.5 vs. 5.1 +/- 0.6 min). However, the relative decrease in the BCECF fluorescence ratio (and therefore pHi) from rest to the fatigue time point was significantly greater (P < 0.05) during the first contractile period (to 65 +/- 4% of initial resting values) compared with the second (77 +/- 4%). The results of the present study demonstrated that, when preceded by an initial fatiguing contractile period, the rise in cytosolic H+ concentration in contracting single skeletal muscle fibers during a second contractile period was significantly reduced but did not attenuate the fatigue process in the second contractile period. These results suggest that intracellular factors other than H+ accumulation contribute to the fall in force development under these conditions.     

Effects of muscle perfusion pressure on fatigue and systemic arterial pressure in human subjects.

The effects of changes in arterial perfusion across the physiological range on the fatigue of a working human hand muscle were studied in seven normal subjects. With the hand above heart level, subjects made repeated isometric contractions of the adductor pollicis muscle at 50% of maximal voluntary contraction in a 6-s on, 4-s off cycle. To assess fatigue, a maximal isometric twitch was elicited in each "off" period by electrical stimulation of the ulnar nerve. The experiment was repeated at least 2 days later with the hand at heart level. Five subjects showed faster fatigue with the arm elevated, and two subjects showed little difference in fatigue for the two conditions. Central blood pressure rose in proportion to fatigue for the subjects overall and returned quickly to its initial level afterwards. We conclude that human muscle fatigue can be increased by physiological reductions in perfusion pressure. Central blood pressure increases as the muscle fatigues, a response that may partially offset declining muscle performance.