Search

Actin/myosin interactions in vertebrate striated muscles are believed to be regulated by the "steric blocking" mechanism whereby the binding of calcium to the troponin complex allows tropomyosin (TM) to change position on actin, acting as a molecular switch that blocks or allows myosin heads to interact with actin. Movement of TM during activation is initiated by interaction of Ca(2+) with troponin, then completed by further displacement by strong binding cross-bridges. We report x-ray evidence that TM in insect flight muscle (IFM) moves in a manner consistent with the steric blocking mechanism. We find that both isometric contraction, at high [Ca(2+)], and stretch activation, at lower [Ca(2+)], develop similarly high x-ray intensities on the IFM fourth actin layer line because of TM movement, coinciding with x-ray signals of strong-binding cross-bridge attachment to helically favored "actin target zones." Vanadate (Vi), a phosphate analog that inhibits active cross-bridge cycling, abolishes all active force in IFM, allowing high [Ca(2+)] to elicit initial TM movement without cross-bridge attachment or other changes from relaxed structure. However, when stretched in high [Ca(2+)], Vi-"paralyzed" fibers produce force substantially above passive response at pCa approximately 9, concurrent with full conversion from resting to active x-ray pattern, including x-ray signals of cross-bridge strong-binding and TM movement. This argues that myosin heads can be recruited as strong-binding "brakes" by backward-sliding, calcium-activated thin filaments, and are as effective in moving TM as actively force-producing cross-bridges. Such recruitment of myosin as brakes may be the major mechanism resisting extension during lengthening contractions.

The accurate measurement of the chemical activators of pain in skeletal muscle has proved to be a major challenge. This study examined the applicability of microdialysis to the measurement of pain-producing substances in skeletal muscle using a defined model of ischemia and reperfusion in the rat. Microdialysis probes were placed into muscle of anesthetized rats. Ischemia was induced for 4 h, followed by reperfusion for 1 h. Perfusates were analyzed for hypoxanthine, potassium, prostaglandin (PG) E(2) and histamine. A 20-fold increase in perfusate hypoxanthine concentration was seen prior to reperfusion (70.1 +/- 27.1 microM for ischemic versus 3.7 +/- 1.9 microM for control; P < 0.05). An initial increase in PGE(2) concentration was seen during ischemia (7.4 +/- 2.0 nM versus 3.4 +/- 1.4 nM; P < 0.05) and immediately post-reperfusion (17.9 +/- 5.2 nM versus 4.0 +/- 1.1 nM; P < 0.05). Potassium concentration was significantly increased following occlusion and reperfusion. This indicates the applicability of microdialysis to the measurement of pain-producing substances in muscle during ischemia and reperfusion. Further use will provide novel information on muscle pain both in defined model systems and in clinical situations in humans., (Copyright 1999 John Wiley & Sons, Inc.)

Information about changes in muscle composition has to date been primarily restricted to histological examination of biopsy samples or qualitative assessment of images obtained using a variety of techniques (e.g., ultrasound, CT, and MRI). We describe the development of a quantitative method for the analysis of muscle composition using MR T2 relaxation time mapping and image analysis. This approach provides an objective means of studying muscle and, when used in conjunction with force production measurements, may provide an accurate measure of response to muscle therapy.

Control C57Bl/10 and mutant, dystrophin-deficient mdx mice of different ages were used to study the permeability of the plasma membrane to cytosolic components, to a vital stain (procion orange) and to extracellular 45calcium. Prenecrotic, 14 +/- 2-day-old mdx mice had normal serum activities of creatine kinase (CK) and pyruvate kinase (PK). Muscles from these animals also had no increased permeability to procion orange or extracellular 45calcium. Serum activities of CK and PK had risen acutely in the 21-day-old mdx mouse compared with control and remained elevated up to 6 months of age. The influx of procion orange and 45calcium content were abnormally elevated in the 40 +/- 4-day-old mdx mouse. These data provide no evidence for an increase in muscle plasma membrane permeability as a primary pathogenic effect of a lack of dystrophin, but results suggest that some factor expressed or de-expressed during mouse development may be necessary for the full expression of the dystrophic process.

Of a total of 225 patients with suspected thoracic outlet syndrome, 37 (16.4%) underwent surgery. Some eight patients required bilateral operations. One patient had a cervical rib and one a prominent C7 transverse process. A total of 45 limbs were operated on. Thirty-nine procedures were performed as combined scalenectomy and transaxillary first rib resection, four as two-stage scalenectomy and transaxillary first rib resection and two as simple scalenectomy alone. Follow-up from 6 to 60 (mean 17) months was available for 39 operations. A two-tier assessment method was used to improve the accuracy of the results of surgery, including the patient's own evaluation of the benefit of operation communicated to an independent observer and the surgeon's clinical appraisal. Assessment of outcome by the physician was excellent in 54%, good in 28%, fair in 10% and 8% had recurrent symptoms. Similar results were achieved in the patients' subjective evaluation with approximately 50% reporting an excellent outcome, about 40% good and 10% fair. A poor result was not recorded in those who underwent combined scalenectomy and transaxillary first rib resection. A radical surgical approach combining scalenectomy and transaxillary first rib resection is advocated to minimize the recurrence rate and improve results.

We have previously discovered a common nonsense mutation in exon 1 of the myophosphosphorylase gene in patients with McArdle's disease, but this failed to explain all cases. We now report a second mutation (G-->TT) in one patient, also in exon 1. This mutation causes a shift in the reading frame which results in the replacement of Val15 by Phe. A further 10 amino acids of mis-sense protein are synthesised before a stop codon is reached.

To study the effect of regeneration on the release of creatine kinase (CK) and prostaglandin E2 from muscle, extensor digitorum longus muscles of mice were injected with 50 microliters of BaCl2 solution in saline (1.2% wt/vol). Injected muscles showed almost complete degeneration at 3 days postinjection but had regenerated to approximately the same fiber cross-sectional area as contralateral control muscles by 12 days postinjection. These muscles released reduced amounts of intracellular CK compared with contralateral control muscles in response to excessive isometric contractile activity or treatment with the calcium ionophore A-23187 (20 microM) in vitro. However, regenerating muscles contained a lower total CK activity than contralateral control muscles, which accounted for the reduced CK efflux after experimental damage. Regenerating muscles released an increased amount of prostaglandin E2 compared with control muscles after both damaging stimuli. We conclude that regenerated muscles show no reduced susceptibility to contraction-induced damage (as assessed by CK release) but show an elevated release of prostaglandin E2 in response to contractile activity or an increase in intracellular calcium.

To examine the role of delay in recovery of peripheral muscle function following exercise in the fatigue experienced by patients with the chronic fatigue syndrome (CFS) and to examine the influence of effort perception in limiting exercise performance in these patients, a study was carried out on a group of twelve patients with chronic fatigue syndrome and 12 sex and age-matched sedentary control subjects. Symptom limited incremental cycle exercise tests including measurements of perceived exertion were performed followed by examination of the contractile properties of the quadriceps muscle group for up to 48 hours. Muscle function was assessed by percutaneous electrical stimulation and maximum voluntary contractions. Muscle function at rest and during recovery was normal in CFS patients as assessed by maximum isometric voluntary contraction, 20:50 Hz tetanic force ratio and maximum relaxation rate. Exercise duration and the relationship between heart rate and work rate during exercise were similar in both groups. CFS patients had higher perceived exertion scores in relation to heart rate during exercise representing a reduced effort sensation threshold of 3.2 units on an unmodified Borg scale in CFS patients. Patients with chronic fatigue syndrome show normal muscle physiology before and after exercise. Raised perceived exertion scores during exercise suggest that central factors are limiting exercise capacity in these patients.

Chronic fatigue syndrome (CFS) is characterized by fatigue at rest which is made worse by exercise. Previous biopsy studies on small numbers of CFS patients have shown a range of morphological changes to which have been attributed fatigue and myalgia. We have now studied 108 patients with CFS or muscle pain and 22 normal volunteers by light and electron microscopy. There was no consistent correlation between symptoms and changes in fibre type prevalence, fibre size, degenerative or regenerative features, glycogen depletion, or mitochondrial abnormalities. Physiological contractile properties of quadriceps (maximal isometric force generation, frequency: force characteristics and relaxation rate) were also examined before and for up to 48 hours after a symptom-limited incremental cycle ergometer exercise test in 12 CFS patients and 12 normal volunteers. Voluntary and stimulated force characteristics were normal at rest and during recovery. Exercise duration was similar in the two groups although CFS patients had higher perceived exertion scores in relation to heart rate during exercise, indicating a reduced effort sensation threshold. On physiological and pathological grounds it is clear that CFS is not a myopathy. Psychological/psychiatric factors appear to be of greater importance in this condition.

The cross-sectional areas of the peroneal and anterior muscle compartments at the same level in the upper leg were measured using magnetic resonance imaging in 41 cases of forefoot pes cavus. The pes cavus group included idiopathic cases and pes cavus associated with Charcot-Marie-Tooth disease, Friedreich's ataxia, cerebral palsy, status postpoliomyelitis, nerve trauma, and spinal cord tethering. Thirty-nine of these cases were symptomatic. The results were compared with studies of 11 normal controls. It was found that in the majority of cases of forefoot cavus, the peroneal compartment was enlarged relative to the anterior compartment when compared with the normal controls. Biopsies of the tibialis anterior and peroneus longus muscles in 18 patients with forefoot pes cavus showed that any relative expansion of the peroneus longus was not due to pseudohypertrophy. Overaction of the peroneus longus in comparison to its antagonist the tibialis anterior is proposed as an important factor in the pathogenesis of the majority of symptomatic cases of forefoot pes cavus.

Magnetic resonance spectroscopy (MRS) and imaging (MRI) are now well established techniques for the study of cellular metabolism and gross structure of muscle. Using non-ferrous materials, we have constructed a system for the measurement of isometric force of quadriceps in response to percutaneous electrical stimulation and voluntary effort within the bore of a 48 cm diameter 1.5 T General Electric SIGNA whole body MR system. Using this system we have been able to study the relationship between electromechanical coupling and chemistry of muscle, with 31P MRS for the measurement of high-energy phosphates and pH, during electrically stimulated activity. Image analysis using the Context Vision system enables a distinction to be made, in 1H MRI by T1/T2 mapping, of muscle, fat and connective tissue to give force per unit cross sectional area of muscle. The combination of MR and functional measurements provide a valuable tool for further detailed analysis of human muscle weakness and fatigue.

1. Accumulation of calcium by extensor digitorum longus muscles from dystrophin-deficient mdx and control C57BL/10 mice has been studied in vitro by measurements of total muscle calcium and by following the retention of 45Ca resulting from the incubation of muscles with the isotope for up to 2 h. 2. The rate of influx of calcium, calculated from the retention of 45Ca, was linear over 2 h in muscles at rest with no significant difference between mdx and control muscles. 3. Repetitive tetanic stimuli caused a substantial increase in 45Ca flux into both mdx and control muscles. This elevated rate of influx was maintained by control muscle, but not by mdx muscle after stimulation resulting in a significantly smaller total calcium flux into mdx muscle compared with control muscle by 1 h after stimulation. Similar changes were also seen in the total muscle calcium content of mdx and control muscles. Comparison of these results with those for loss of cytosolic creatine kinase previously reported (McArdle, A., Edwards, R.H.T. & Jackson, M.J. Clin. Sci. 1991; 80, 367-71) [1] indicate that control and dystrophin-deficient muscles release equivalent amounts of intracellular creatine kinase in response to the same accumulation of intracellular calcium. 4. These results therefore do not support the hypotheses that dystrophin deficiency in muscle leads to increased calcium influx during contractile activity, or that dystrophin-deficient muscle shows any inherent increased permeability to cytosolic proteins.

Creatine kinase (CK) release in response to excessive electrically stimulated contractile activity has been studied in isolated rat soleus muscles. The exacerbation of CK release induced by contractile activity was found to be directly related to the length of time for which the muscle was stimulated and indirectly related to the recovery of force following the end of stimulation. 31P-NMR studies were undertaken using a recirculating superfused muscle preparation and demonstrated that muscles subjected to two different stimulation protocols (stimulation for 0.5 s every 2 s in oxygenated medium or for 1.5 s every 2 s in anoxic medium) had similar falls in ATP content and pH despite a substantially greater release of CK from the muscles stimulated under anoxia. However, stimulated muscles under anoxia showed a more rapid fall and reduced recovery of phosphocreatine and a greater sustained elevation of inorganic phosphate than muscles in oxygenated medium. It is concluded that only part of the increased loss of CK from muscles stimulated in anoxic medium can be explained by release from cells which have lost energy supplies and therefore that other mechanisms must exist which allow release of CK and other cytosolic enzymes from muscle cells.

Previous studies have demonstrated that supplemental alpha-tocopherol inhibited calcium-induced cytosolic enzyme efflux from normal rat skeletal muscles incubated in vitro and suggested that the protective action was mediated by the phytyl chain of alpha-tocopherol [1]. In order to investigate this further a number of hydrocarbon chain analogues of tocopherol (7,8-dimethyl tocol, 5,7-dimethyl tocol, tocol, alpha-tocotrienol, alpha-tocopherol [10], vitamin K1, vitamin K1 [10], vitamin K1 diacetate, vitamin K2 [20], phytyl ubiquinone and retinol) were tested for any ability to inhibit calcium ionophore, A23187, induced creatine kinase (CK) enzyme efflux. Some compounds were found to be very effective inhibitors and comparison of their structures and ability to inhibit TBARS production in muscle homogenates revealed that the effects did not appear related to antioxidant capacity or chromanol methyl groups, but rather the length and structure of the hydrocarbon chain was the important mediator of the effects seen.

Patients with postviral fatigue syndrome (PVFS) usually complain of the skeletal muscle-related symptoms of fatigue and myalgia. It is not surprising therefore that the muscles have recently been the object of intensive studies which have used a variety of biochemical and physiological techniques. The aim of this chapter is to review these findings, and to discuss their significance or otherwise to the presenting symptoms and course of the condition.

1. The release of glutathione has been studied in comparison with the release of creatine kinase from isolated rat soleus muscles subjected to certain forms of experimental damage. 2. Excessive electrically stimulated contractile activity or treatment of muscles with the mitochondrial inhibitor, 2,4-dinitrophenol, induced a substantial release of both creatine kinase and glutathione and a reduction in the total glutathione content of the muscle. The time course of this release and depletion indicates that the efflux of the two molecules is not directly related and that a reduction in muscle glutathione content does not occur before cytosolic enzyme release. 3. 2,4-Dinitrophenol-stimulated release of creatine kinase was significantly reduced by the omission of external calcium from the incubation media, but glutathione release and depletion was relatively unaffected by this. Deliberate elevation of the muscle intracellular calcium content with the calcium ionophore, A23187, induced a substantial loss of creatine kinase, but had no significant effect on the release of glutathione. 4. Muscle biopsies from patients with Duchenne muscular dystrophy were found to have an elevated content of glutathione and an equivalent protein-thiol content compared with control subjects. 5. We conclude that, although release of glutathione from skeletal muscle occurs after excessive contractile activity or inhibition of mitochondrial metabolism, this is not a key step in the damaging processes leading to cytosolic enzyme release, neither is it relevant to the ongoing damage to skeletal muscle which occurs in patients with Duchenne muscular dystrophy.

1. mdx mice do not express dystrophin, the product of the gene which is defective in Duchenne and Becker muscular dystrophy. We have previously shown that protein-synthetic rates (ks) are increased in mdx mouse muscles [MacLennan & Edwards (1990) Biochem. J. 268, 795-797]. 2. The tumour-promoting stereoisomer of phorbol 12,13-didecanoate (4 beta-PDD) acutely increased the ks of muscles from mdx and wild-type (C57BL/10) mice incubated in vitro in the absence of insulin. The effects of 4 beta-PDD are presumably mediated by activation of protein kinase C (PKC). 3. The muscle glycogen concentrations of mdx mice were higher than those of C57BL/10 mice. Studies performed in vivo and in vitro suggested that the effect might be at least partially due to increased rate of glycogen synthesis in mdx muscle. 4. 4 beta-PDD increased the glycogen-synthetic rates rates of C57BL/10, but not mdx, muscles incubated in vitro in the absence of insulin. 5. In muscles from both species incubated in the absence of insulin, treatment with 4 beta-PDD also induced increased rates of glucose uptake and lactate production. Kinetic studies of C57BL/10 and mdx muscles suggested that 4 beta-PDD raised the Vmax. of glucose uptake, but did not alter the Km for the process. 6. The possible role of PKC in controlling the protein and carbohydrate metabolism of normal and mdx mouse muscles is discussed.

Mdx mice have a genetic defect similar to that which causes Duchenne muscular dystrophy in humans. The influence of calcium on muscle protein metabolism of mdx and wild type (C57BL/10) mice was examined in vitro. Incubation of mdx muscles in a medium containing calcium at a concentration of 2.0 mM (but not 0.2 mM) resulted in proteolytic rates that were greater than those of C57BL/10 muscles. At 2.0 mM extracellular calcium, mdx muscle proteolysis was attenuated by thiol protease inhibitors but not by the weak base methylamine. Protein synthetic rates were higher in incubated mdx muscles than in incubated C57BL/10 muscles, but no effect of extracellular calcium concentration was observed in either strain. These data suggest that mdx mice have an abnormality of muscle calcium handling, which results in activation of nonlysosomal proteolytic processes but does not exert acute effects on protein synthetic rate.

1. Isolated extensor digitorum longus muscles from control C57BL/10 and mutant dystrophin-deficient C57BL/10 mdx mice have been studied in vitro to determine whether dystrophin deficiency influences the susceptibility of muscle to contractile activity-induced damage. 2. mdx muscles were found to release reduced amounts of intracellular creatine kinase compared with control tissue in response to excessive contractile activity with or without simultaneous stretching of the muscle to 130% of its resting length. 3. In contrast, prostaglandin E2 release from mdx muscle was elevated compared with control tissue in response to either form of contractile activity or to treatment with the calcium ionophore A23187. 4. These results do not support the hypothesis that dystrophin-deficient muscle is more susceptible to damage induced by contractile activity, but suggest that dystrophin deficiency influences the activity of muscle membrane phospholipase enzymes.

Sodium dodecyl sulfate-polyacrylamide gel analysis of the incubation media surrounding isolated rat soleus muscles has been used to determine the nature of the proteins lost from skeletal muscle during damage. Following various forms of experimental trauma only selected proteins are lost to the medium when compared to the protein composition of the muscle cytosol. The proteins released do not appear to be released in a size dependent manner and their release is partly inhibited by an absence of calcium in the extracellular medium. alpha-Tocopherol entirely prevents the release of cytosolic enzymes induced by intracellular calcium overload.

We studied the release of creatine kinase (CK) activity and prostaglandin E2 (PGE2) from isolated strips of biceps muscle from patients with Duchenne muscular dystrophy and nondystrophic control patients. CK release was significantly higher from the dystrophic samples than controls during the initial period of incubation, but this difference reduced with time of incubation. Immediate immersal of muscle strips into calcium-free bathing medium reduced the initial difference between the efflux from dystrophic and nondystrophic samples, whereas treatment with the calcium ionophore maintained the difference between the groups throughout the period of incubation (150 minutes). These results support the hypothesis that the lack of dystrophin in Duchenne muscle leads to damage to the tissue via a failure of calcium homeostasis. PGE2 release from the muscle strips followed a similar pattern to CK activity, supporting the possibility that, at least partly, the calcium-mediated damage involves activation of phospholipid hydrolysis.

This paper considers an internal standard of lumbar muscularity. The cross-sectional areas (Acs) of the intervertebral disc and paraspinal muscles were measured in 147 working men from an axial magnetic resonance image passing through the L3-4 disc. Lumbar muscularity was expressed by two ratios; the ratio between the Acs of the right psoas and the Acs of the intervertebral disc (P:disc), and the ratio between the combined Acs values of the right erector spinae and quadratus lumborum and the Acs of the disc (ESQL:disc). When the subjects were divided into two age groups (76 aged 20-30 years and 71 aged 31-58 years) lumbar muscularity was found to be significantly greater (P less than 0.001) in the younger age group (P:disc = 0.8, SD 0.2; ESQL:disc = 2.0, SD 0.3) than in the older age group (P:disc = 0.7, SD 0.2; ESQL:disc = 1.8, SD 0.3). Lumbar muscularity was not significantly affected by occupation or by a history of low back pain.

(1) A recirculating isolated superfused skeletal muscle preparation has been developed for the study of rat soleus muscles at physiological temperature using 31P Nuclear Magnetic Resonance (NMR). (2) This system has been used to study intracellular muscle high energy phosphate content and pH during experimental damage to the muscle induced by 2,4-dinitrophenol, deoxycholate and the calcium ionophore, A23187. (3) Results indicate that release of intracellular cytosolic enzymes from damaged skeletal muscle may be induced by phosphocreatine (PCr) and adenosine trisphosphate (ATP) depletion, but under certain circumstances intracellular enzymes can be released from skeletal muscle without any fall in muscle PCr or ATP content.

Damage to skeletal muscles may be mediated via free radicals or intracellular calcium overload. To look for inter-relationships between these pathways we have examined the effect of intracellular Ca overload on muscles from rats fed on either a vitamin E-deficient or vitamin E-sufficient diet and assessed the non-enzymic lipid peroxidation in these muscles by examining the production of thiobarbituric acid reactive substances by homogenates. Vitamin E-deficient muscles were more susceptible to Ca-induced intracellular enzyme efflux and this was acutely corrected by supplementation of the external medium with 230 mumol alpha-tocopherol/l. Vitamin E-deficient muscles showed increased levels of basal lipid peroxides and were more susceptible to iron-catalysed lipid peroxidation. Addition of the Ca ionophore A23187 increased lipid peroxidation in vitamin E-deficient muscle homogenates, but had the opposite effect in vitamin E-sufficient muscles. These results demonstrate that vitamin E-deficient muscle has an increased susceptibility to intracellular Ca overload, but that this effect cannot be explained by a direct stimulatory effect of the ionophore on non-enzymic lipid peroxidation.

mdx mice lack the protein dystrophin, the absence of which causes Duchenne muscular dystrophy in humans. To examine how mdx mice maintain muscle mass despite dystrophin deficiency, we measured protein turnover rates in muscles of mdx and wild-type (C57BL/10) mice in vivo. At all ages studied, rates of muscle protein synthesis and degradation were higher in mdx than in C57BL/10 mice.

In muscular dystrophy and metabolic myopathy there is wasting and replacement of the muscle bulk with fat and fibrous tissue. A method for estimating the fat content of muscle and the cross-sectional area from computerised tomography (CT) scans is presented. Data from the quadriceps muscle of fifty patients show the effect of fat replacement on muscle strength. The CT scans have shown that the pattern of wasting in individual muscle groups is characteristic of the type of myopathy. In Duchenne muscular dystrophy, hypertrophic and pseudohypertrophic muscle have hitherto been indistinguishable, but this method allows a clear distinction to be made. Quantitative CT offers a new approach to the investigation of muscle diseases and can also be of considerable value in selecting sites for needle biopsy of muscle.

The ability of muscle fibres to sustain force can be related to their economy of energy utilization and to their capacity to regenerate energy under the prevailing conditions (aerobic or anaerobic) of contraction. The pathophysiology of muscle fatigue is analysed in patients with thyroid dysfunction and with impaired glycogenolysis, and in a patient with abnormal mitochondrial function. Muscle from hypothyroid patients, like cooled muscle, is slow in relaxing and shows a reduced energy requirement (energy economy) and reduced fatiguability, whereas muscle of hyperthyroid patients may show the opposite features. In myophosphorylase deficiency the energy economy is normal in the fresh state and increases as contraction proceeds; however, fatigue is premature and associated with impaired excitation rather than an overall depletion of energy stores. With abnormal mitochondrial function the muscle tends to be effectively anaerobic and fatigue is associated with impaired excitation-contraction coupling. This appears to result from either muscle ischaemia or the dominant use of anaerobic metabolism for energy regeneration. Fatigue in these disorders of energy metabolism may ultimately be due to a reduced supply of ATP but direct evidence of this is lacking and, if it occurs, its physiological expression is probably variable.

Muscle pain is the presenting symptom in patients with a wide variety of conditions. Such patients often pose problems in diagnosis and management and a rational scheme for their investigation is needed. The results of muscle biopsy, electromyography, exercise and strength testing and blood measurements in 109 consecutive patients presenting with myalgia are reported. By determining the sensitivity and specificity of the tests in identifying patients with specific muscle abnormalities, a rational investigative protocol has been constructed. Creatine kinase and erythrocyte sedimentation rate are the most useful screening tests and if either is abnormal, muscle biopsy, electromyography, muscle strength and exercise testing are then performed. Despite exhaustive investigation, specific muscle abnormalities were found in only one-third of the patients; whilst many of the remaining patients undoubtedly had a psychogenic component to their pain, it is likely that a number of unidentified specific metabolic defects remain as causes of myalgia.

To examine fatigue mechanisms in an unselected series of patients with excess fatigue ("effort syndromes") their muscle function was compared with that of normal subjects. Voluntary performance was assessed with a cycle ergometer to exhaustion and by maximal isometric contractions of the quadriceps femoris. The mean maximal heart rate in patients during ergometry was 89% of the predicted rate, and quadriceps strength was either normal or was inappropriate for the available muscle, which suggested submaximal effort. Contractile performance was examined in the absence of volition with stimulated contractions of the adductor pollicis. During stimulated fatiguing activity patients were neither weaker nor more fatigable than controls; thus the excess fatigue experienced by the patients was not due to a defect of the contractile apparatus. The increased perception of effort must therefore be due to impairment of central rather than peripheral mechanisms. The optimal approach to treatment of effort syndromes combines physical and psychological techniques.

The relaxation rate from electrically stimulated isometric contractions of human adductor pollicis has been measured during rewarming following cooling, during ischaemia and during ischaemic contractile activity. The Q10 for relaxation rate (25-37 degrees C intramuscular temperature) was estimated as 2 . 3. Relaxation rate was found to decline at a rate of 1 . 5% initial value per minute of ischaemia. Relaxation rate declined more rapidly during ischaemic contractile activity than during ischaemia alone. The rate of decline was more closely related to the force X time performed than the number of excitatory impulses. During a supramaximal 20 Hz tetanus, relaxation rate declined markedly at a time when only slight force fatigue had occurred; hence the decline could not be explained by selective fast muscle fibre fatigue. No recovery occurred during ischaemic rest following ischaemic activity but, following restoration of the circulation, recovery occurred with a half time of one minute being virtually complete at 5-7 min. Changes in relaxation rate during fatiguing contractions and recovery from them follow different time courses from muscle excitability and force production. Change in relaxation rate cannot be simply related to changes in the concentration of major energy metabolites. It is proposed that relaxation rate is related to the rate of energy turnover in the contracting muscle.

Ischaemic muscle pain was studied in a total of 78 experiments on 3 normal subjects. The development of pain induced by intermittent voluntary isometric contractions was recorded whilst force and frequency of contraction were varied. As the muscle progressively expends energy, pain (P) develops in an exponential fashion and can be expressed by the function P = (F x t)k/C where (F x t) is the integral of force with time, k, an exponent, and C, a constant. The exponent is fairly constant despite changes in frequency. The time of pain onset and energy expenditure at the point of intolerable pain depend on frequency. At a given frequency, energy expenditure at intolerable pain is independent of the force of contraction.

The calcium, magnesium, sodium and potassium content of needle biopsy samples of human muscle from control subjects and patients with various neuromuscular disorders has been analysed by atomic absorption spectrometry. The calcium content of muscle from both patients with Duchenne Muscular Dystrophy and other myopathies was significantly elevated compared to both normal and patient control subjects. Muscle sodium content was not significantly altered in any group compared to control subjects. Decreased magnesium and potassium levels were found in muscle from patients with Duchenne Muscular Dystrophy, although the muscle magnesium/potassium ratio was unchanged, the most likely explanation for this being non-specific loss of intracellular components from the cell.

Seventeen volunteers performed unilateral strength-training of the quadriceps with high-resistance, low-repetition, dynamic exercise, thrice weekly for an average of 5 weeks. Both before and after the training period, bilateral measurements were made of isometric quadriceps strength, quadriceps cross-sectional area (by ultrasound scanning), and thigh circumference. There were no significant changes in the untrained thighs. The trained quadriceps increased their isometric strength by more than they changed their cross-sectional area (mean increments = 15% and 6% respectively). Quadriceps hypertrophy was underestimated by measurements of thigh circumference and could not be predicted from them. We conclude that studies of localized muscle growth require direct measurements of the size of the muscle(s) concerned. Nevertheless, these may still underestimate the improvements in strength produced by high-resistance training.

1. After severe muscular contraction in man recovery of force is largely complete in a few minutes, but is not wholly so for many hours. The long-lasting element of fatigue is found to occur primarily for low frequencies of stimulation (e.g. 20/sec), and is much less pronounced, or absent, at high frequencies (80/sec). The twitch force is an unreliable measure of the state of fatigue. 2. The long-lasting element of fatigue is not due to depletion of high-energy phosphate nor is it due to failure of electrical activity as recorded from surface electrodes. It is probably the result of an impairment of the process of excitation-contraction coupling. Its practical importance for man could be significant as an explanation of the subjective feelings of weakness following exercise.

1. The force produced by isometric contractions of the quadriceps muscle have been studied during maximal voluntary contractions and when a substantial part of the muscle was electrically stimulated via surface electrodes. 2. In normal children and adults, the force of a maximal voluntary contraction of the quadriceps was proportional to body weight. 3. The function of the quadriceps has been described in terms of the force/frequency curve, speed of relaxation and the rate of loss of force during 18 s stimulation at 30 Hz and 100 Hz. 4. The functional characteristics of adductor pollicis when stimulated via the ulnar nerve were essentially similar to those of the quadriceps. 5. Studies of the function of these two muscles are complementary since quadriceps femoris is amenable to needle biopsy investigations of its structure and chemistry whereas adductor pollicis is more suitable for electrophysiological studies.

1. A technique has been devised to measure the effect of stimulation frequency on the contraction force in an accessory muscle of respiration, the sternomastoid, in man. The frequency/force curve was found to be very similar to that of the quadriceps and adductor pollicis muscles. 2. Fatigue of the sternomastoid due to inspiratory loading or sustained maximum voluntary ventilation resulted in reduced force generation at low stimulation frequencies compared with maximum force. This type of fatigue frequently persisted for several hours. 3. If low frequency fatigue were to develop in patients with pulmonary disease it could have important consequences for the development of respiratory failure.

Needle biopsies from vastus lateralis in untreated hypothyroid patients with muscle weakness confirmed by quadriceps force measurements (n = 11) were repeated when the patients had taken L-thyroxine for a mean period of 9.2 months (range 5.3-13.3 months, n = 8) and had been continuously biochemically euthyroid for a mean period of 4.9 months (range 2-11 months). Biopsies were analysed biochemically for mitochondrial function. On light microscopy, histochemical examination, mean fibre areas and fibre percentages of type I and type II fibres were determined. Electronmicroscopy was also performed. Abnormalities on light microscopy occurred in eight patients of which type II fibre atrophy was the commonest and of the remainder two patients showed a myopathic electromyogram (EMG) and a raised plasma creatine kinase activity and one ultrastructural change on biopsy. After treatment resolution of pathological changes was often slow and half the patients had persistent abnormalities when rebiopsied. The type I mean fibre area was significantly increased in the eight hypothyroid females (p less than 0.05) and type II mean fibre areas tended to be low and in females this was significant (p less than 0.05). After treatment the type I mean fibre area was significantly reduced (p = 0.05). The type II mean fibre area also tended to fall but this was not significant (p greater than 0.05). No change in the fibre percentages occurred. A myopathic EMG, a raised plasma creatine kinase activity, ultrastructural changes and low mitochondrial enzyme activities on needle biopsy were other common findings and their significance is discussed.

An isolated skeletal muscle preparation is described which has been used to study the efflux of enzymes in response to contractile activity, metabolic poisons and detergent treatment. In both fast and slow muscles contractile activity caused a release of lactate dehydrogenase and creatine kinase that reached a peak 1-2 h after the end of stimulation. There was very little release during the 30 min stimulation period whether the muscles were under aerobic or hypoxic conditions. Incubation of the muscles with cyanide and iodoacetate caused a similar delayed release of enzyme. Disruption of cell membranes with detergent treatment caused an entirely different and very rapid pattern of enzyme release. Enzyme release from the fatigued isolated muscle preparations appears to be initiated as a consequence of phosphorylcreatine or ATP depletion. The relevance of this to release of muscle enzymes after activity in vivo is discussed.

The light microscopic appearance of the human tibialis anterior muscle is described based on conchotome biopsy specimens from seven healthy volunteers and 20 patients who presented with myalgia but who had no evidence of neuromuscular disease. The morphometric characteristics of these normal muscles are documented and the similarities and differences between the appearances of the tibialis anterior and other muscles discussed.

Ultrasound techniques have been used for the non-invasive, quantitative characterisation of muscle tissue in normal subjects and volunteer patients. Radio frequency (RF) echoes from a volume of tissue have been digitised and analysed using computer techniques. Attention has been given to the correct positioning and orientation of the transducer during examination due to the importance of the angle dependence of the interaction of ultrasound with muscle fasciculi. Several different muscles in the leg, arm and back of normal subjects have been examined, whereas patient studies have concentrated on the vastus intermedius. Ultrasonic data from patients with muscular dystrophy have been correlated with measurements of muscle tissue density obtained using X-ray computerised tomography (X-ray CT). The technique shows that ultrasound can be used to differentiate between normal and diseased muscle quantitatively. Results indicate that pathological change can be detected and monitored earlier with ultrasound than with X-ray CT. These quantitative methods are now in use as a guide to the staging and monitoring of pathological change in muscle.

Plasma pyruvate kinase (EC 2.7.1.40) activity contains two major isoenzymes. The "L" type originates from liver (erythrocytes contain a similar isoenzyme), the "M" type predominantly from skeletal muscle. The diagnostic use of pyruvate kinase as an index of muscle leakage requires an assay of only the M type. An M-type-selective assay for mixtures of the two isoenzymes was devised, based on the differences that the L form has a higher Km for phosphoenolpyruvate and is inhibited by about 95% by a high concentration of ADP. All relevant conditions of the assay have been examined and the reaction system has been optimized at 30 degrees C. The assay yields the expected Km values for M type pyruvate kinase. An apparent increase in activity in hemolysed plasma samples was found to be derived from leukocytes.

Abnormalities in skeletal muscle function were characterized in 19 hypothyroid subjects and the extent and time course of recovery in these abnormalities after treatment determined. Slow muscle relaxation was observed on examination in 11 patients and 12 out of 16 patients had a reduced quadriceps maximum relaxation rate (MRR) and 11 out of 15 had a prolonged ankle jerk relaxation time (AJRT). Measurements of quadriceps force revealed significant muscle weakness when comparison with 15 age matched healthy subjects was made (P less than 0.001). This weakness was often not evident on clinical examination. A raised plasma creatine kinase (CK) activity was characteristic. With treatment the plasma CK activity and the AJRT rapidly became normal often long before the serum thyroid-stimulating hormone (TSH) was normal, but the quadriceps MRR was slower to recover. It was rarely normal before the serum TSH became normal. Weakness was slow to recover and seven patients remained weak at the end of the study despite being euthyroid for a mean period of 1 year, but strength increased modestly overall (P less than 0.01). The urinary creatinine/height index was unchanged, indicating that total muscle mass does not alter with therapy.