Isometric skeletal muscle force measurement in primary myopathies

Introduction: In myopathy patients, it is useful to measure skeletal muscle forces. Conventional methods require voluntary muscle activation, which can be unreliable. We evaluated a device for nonvoluntary force assessment. Methods: We tested 8 patients (unknown myopathy n = 2, inflammatory myopathy, facioscapulohumeral muscular dystrophy, mitochondrial myopathy, dysferlinopathy, multi‐minicore disease, Becker‐Kiener muscular dystrophy, n = 1 each). Isometric twitch torques of ankle dorsiflexors were measured after fibular nerve stimulation. Results: Six patients had decreased torques vs. 8 controls (men: median Newton‐meter 1.6 vs. 5.7, women: 0.2 vs. 3.9, both P < 0.0001). Values correlated with Manual Muscle Test results (r = 0.73; r² = 0.53; P < 0.0001). In weak dorsiflexors, torque could be measured despite lower signal‐to‐noise ratios. In 2 patients with hypertrophy, we measured increased torques. Conclusions: Nonvoluntary muscle force assessment can be used in patients with myopathies, and values correlate with voluntary forces determined by traditional methods. Muscle Nerve 53 : 913–917, 2016     

Effects of alcohol on skeletal and cardiac muscle

The acute and chronic toxic effects of alcohol on skeletal and cardiac muscle are clinically important. Muscle weakness and atrophy are the main manifestations of skeletal myopathy, and arrhythmias and progressive left-ventricular dysfunction are those of cardiomyopathy. Most patients remain asymptomatic from these effects for a long time. Myocyte atrophy and death are the main pathological findings. A clear dose-related effect has been established with ethanol consumption, with gender and some specific gene polymorphisms being factors of increased susceptibility to alcohol-induced muscle damage. Pathogenic mechanisms are pleiotropic, the most relevant being disturbances in carbohydrate, protein, and energy cell turnover, signal transduction, and induction of apoptosis and gene dysregulation. Ethanol abstinence is the only effective treatment, although controlled drinking is useful in patients who do not achieve abstinence. Persistent high-dose consumption results in deterioration of muscle and heart function, with a high mortality due to arrhythmias and progression of heart failure.     

The effects of strength,aerobic, and concurrent exercise on skeletal muscle damage in rats

Introduction: In this study we examined oxidative stress and skeletal muscle damage resulting from acute strength, aerobic, or concurrent exercise in rats. Methods: The animals were divided into control (C), strength (SE), aerobic (AE), and combined (CE) exercise groups. They were euthanized at 3 different time‐points (6, 24, and 48 h) after acute exercise. Results: SE exercise rats had increased dichlorofluorescein oxidation at 6 h post‐exercise and decreased superoxide dismutase activity at all time‐points. Glutathione peroxidase activity and sulfhydryl levels were increased in the AE group at 48 h post‐exercise. Serum lactate dehydrogenase activity was increased in the SE and CE groups at 24 h and in the AE group at 48 h. Echo intensity was elevated at 24 h for all groups. Conclusions: Forty‐eight hours was sufficient for complete recovery from oxidative stress and muscle damage in the SE and CE groups, but not in the AE group. Muscle Nerve 50 : 79–86, 2014     

Mitochondria of trained skeletal muscle are protected from deleterious effects of statins

Introduction: Statins are associated with adverse skeletal muscle effects. Our objective was to determine if muscular adaptations following exercise training prevented deleterious effects of atorvastatin in glycolytic skeletal muscle. Methods: Twenty rats were divided into 2 groups: a control group (n = 10; Cont) and a 10 days of training group (n = 10; Training). Using the permeabilized fibers technique, we explored mitochondrial function. Results: Exercise training increased V(max) and H(2) O(2) production without altering the free radical leak, and mRNA expression of SOD2 and Cox1 were higher in trained muscle. In the Cont group, atorvastatin exposure increased H(2) O(2) production and decreased skeletal muscle V(max) . The decreased V(max) effect of atorvastatin was dose dependent. Interestingly, the half-maximal inhibitory concentration (IC(50) ) was higher in the Training group. H(2) O(2) production increased in trained muscle after atorvastatin exposure. Conclusions: These results suggest that improvements in mitochondrial respiratory and antioxidant capacities following endurance training protected mitochondria against statin exposure. Muscle Nerve 46: 367-373, 2012.     

Calibrated quantitative ultrasound imaging of skeletal muscle using backscatter analysis

We evaluated the ability of an ultrasound method, which can characterize cardiac muscle pathology and has reliability across different imaging systems, to obtain calibrated quantitative estimates of backscatter of skeletal muscle. Our procedure utilized a tissue-mimicking phantom to establish a linear relationship between ultrasound grayscale and backscatter levels. We studied skeletal muscles of 82 adults: 45 controls and 37 patients with hereditary myopathies. We found that skeletal muscle ultrasound backscatter levels varied with probe orientation, age, and muscle contraction and pathology. Reliability was greater with the probe in longitudinal compared with transverse planes. Backscatter levels were higher in those >40 years of age, in muscle extension than flexion, and in myopathic patients than controls. Calibrated measurements of muscle backscatter have sensitivity and specificity in identifying and reliably measuring levels of skeletal muscle pathology.     

Clinical heterogeneity associated with mitochondrial DNA depletion in muscle

We studied 10 patients with a variable degree of mtDNA depletion in muscle. Seven patients showed a clear-cut myopathic pattern, while the three remaining had brain involvement. There was no relationship between age at onset and relative mtDNA copy number in muscle, but we found an apparent correlation between clinical severity and degree of muscle mtDNA depletion. Muscle morphology showed that mtDNA depletion was associated with mitochondrial proliferation and cytochrome c oxidase negative fibers. Biochemical studies revealed single or combined defects of mtDNA-dependent respiratory chain complexes. Our data indicate that patients with mtDNA depletion may have a more variable age at onset and clinical evolution and wider phenotype than previously thought. The diagnosis of this condition, so far regarded as rare, may have been overlooked to some extent.     

Contractility of mdx skeletal muscle after denervation and devascularization.

To evaluate the regenerative capacity of mdx skeletal muscle, changes in contractile properties of the fast-twitch extensor digitorum longus (EDL) of normal and mdx mice were studied at 7 and 16 weeks of age, following denervation and devascularization (DD) at 4 weeks of age. At 7 weeks, DD EDL of both strains showed significantly decreased isometric twitch and tetanus tensions compared with their non-DD controls. By 16 weeks, normal operated muscle exhibited a recovery of 57% and 58% of absolute tetanus and twitch tensions while the mdx EDL recovered remarkably to 96% and 99% of 7-week values. At 7 weeks, the DD EDL of both strains exhibited significantly slower time-to-peak (TTP) and one-half relaxation time (1/2RT). By 16 weeks, TTP and 1/2RT of the mdx DD EDL no longer differed from non-DD controls, but the normal EDL showed slowed TTP. No differences were found in the maximum velocity of shortening (Vo) or in posttetanic potentiation (PTP). Following DD, there was an increase in resistance to fatigue in both strains at 7 weeks. This resistance persisted at 16 weeks in the normal mouse, but the operated mdx EDL returned to normal. It would appear that following a denervation/devascularization insult, the mdx EDL is able to recover contractile characteristics to a remarkably larger extent than normal EDL.     

Demonstration of mitochondrial ribosomal RNA in frozen and paraffin–embedded sections of skeletal muscle by in situ hybridization

We have used in situ hybridization with synthetic oligonucleotide probes to the 16S sub–unit of mitochondrial ribosomal RNA to detect mitochondrial accumulations in biopsies of skeletal muscle from 16 patients with mitochondrial myopathies. In all cases, the distribution of the hybridization signal matched the pattern of succinate dehydrogenase activity in adjacent cryostat sections. In situ hybridization also allowed visualization of mitochondrial accumulations in paraffin–embedded sections of skeletal muscle that had been fixed in either De Castro's solution or formalin. DNase pre–treatment did not significantly diminish the intensity of hybridization signal, suggesting that the hybridization is predominantly to RNA. The advantages of this technique are that it allows the investigation of mitochondrial disorders when only paraffin–embedded tissue is available and should also facilitate the demonstration of mitochondrial accumulations in tissues other than skeletal muscle.     

骨骼肌细胞线粒体通透性转换孔及bcl-2和bax mRNA表达与运动

背景：运动影响骨骼肌细胞的凋亡,而线粒体途径是介导细胞凋亡的一个重要途径。 目的：研究运动对大鼠骨骼肌线粒体通透性转换孔、凋亡调控基因bcl-2和bax表达的影响。 方法：将24只成年雄性SD大鼠随机分为3组：对照组正常饲养,6周游泳训练组进行6周的游泳训练,每周6次,一次性游泳力竭组于第6周进行一次力竭性游泳运动。应用紫外分光光度仪检测各组大鼠骨骼肌线粒体通透性转换孔的开放情况,应用RT-PCR测定大鼠骨骼肌bcl-2和bax mRNA的表达。 结果与结论：与对照组比较,6周游泳训练组大鼠骨骼肌线粒体通透性转换孔的开放程度变化不明显,bcl-2 mRNA的表达显著增加,bax mRNA的表达显著减少,bcl-2/bax mRNA比值显著增大(P < 0.01)。与对照组比较,一次性游泳力竭组大鼠骨骼肌线粒体通透性转换孔开放程度明显增加(P < 0.01),bcl-2 mRNA的表达显著减少,bax mRNA的表达显著增加,bcl-2/bax mRNA比值显著减小(P < 0.01)。说明运动训练可通过改变线粒体通透性转换孔的开放、调节bcl-2/bax表达,调控骨骼肌细胞凋亡。     

Statin-associated myopathy and its exacerbation with exercise

3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) are a common and effective treatment for hypercholesterolemia, with a low overall rate of side-effects. The most common complication is some degree of skeletal muscle myopathy, ranging from painless serum creatine kinase elevations to rhabdomyolysis. Unfortunately, the likelihood and/or severity of complications increases with the combination of statin treatment and physical activity. The specific pathways that mediate statin-associated myopathy are unclear, and research directly addressing the exacerbation with exercise is limited. Potential mechanisms include the induction of skeletal muscle fiber apoptosis, alterations in ubiquitin-proteasome pathway activity, mitochondrial dysfunction, and terpenoid depletion. In this review we provide an overview of research that specifically addresses the combination of statin-associated myopathy and physical activity and highlight some deficiencies in the available literature, as well as future directions for this important subset of statin-associated myopathy.     

Relation between in vivo and in vitro measurements of skeletal muscle oxidative metabolism.

The relationships between in vivo (31)P magnetic resonance spectroscopy (MRS) and in vitro markers of oxidative capacity (mitochondrial function) were determined in 27 women with varying levels of physical fitness. Following 90-s isometric plantar flexion exercises, calf muscle mitochondrial function was determined from the phosphocreatine (PCr) recovery time constant, the adenosine diphosphate (ADP) recovery time constant, the rate of change of PCr during the initial 14 s of recovery, and the apparent maximum rate of oxidative adenosine triphosphate (ATP) synthesis (Q(max)). Muscle fiber type distribution (I, IIa, IIx), citrate synthase (CS) activity, and cytochrome c oxidase (COX) activity were determined from a biopsy sample of lateral gastrocnemius. MRS markers of mitochondrial function correlated moderately (P < 0.05) with the percentage of type IIa oxidative fibers (r = 0.41 to 0.66) and CS activity (r = 0.48 to 0.64), but only weakly with COX activity (r = 0.03 to 0.26, P > 0.05). These results support the use of MRS to determine mitochondrial function in vivo.     

Skeletal muscle disorders associated with selenium deficiency in humans

Skeletal muscle disorders manifested by muscle pain, fatigue, proximal weakness, and serum creatine kinase (CK) elevation have been reported in patients with selenium deficiency. The object of this report was to review the conditions in which selenium deficiency is associated with human skeletal muscle disorders and to evaluate the importance of mitochondrial alterations in these disorders. A systematic literature review using the Medline database and Cochrane Library provided 38 relevant articles. The main conditions associated with selenium deficiency fell into three categories: (1) insufficient selenium intake in low soil-selenium areas; (2) parenteral or enteral nutrition, or malabsorption; and (3) chronic conditions associated with oxidative stress, such as chronic alcohol abuse and human immunodeficiency virus (HIV) infection. In low soil-selenium areas, reversibility of muscle symptoms was similar after selenium supplementation and placebo administration, suggesting a role for other factors in the development of disease. In parenteral or enteral nutrition, or malabsorption, muscle symptoms improved after selenium supplementation in 18 of 19 patients (median delay: 4 weeks). The reason that only a minority of selenium-deficient patients present with skeletal muscle disorders is unclear and is possibly related to cofactors, such as viral infections and drugs. Prospective studies of selenium-deficient myopathies would be useful in critically ill patients, alcohol abusers, and HIV-infected patients.     

Effect of neural activity on skeletal muscle phosphoproteins

A number of phosphoproteins were found in the soluble fraction of rat skeletal muscle by two-dimensional polyacrylamide gel electrophoresis (PAGE). The pattern of some of these phosphoproteins differed in fast (extensor digitorum longus, EDL) or slow (soleus) muscles, was dependent on normal innervation, and was altered with denervation. In order to determine if the pattern was maintained by electrical activity or trophic factors, we compared the effect of electrical block by local neural application of tetrodotoxin with the effect of complete nerve section. Both methods produced similar alterations in phosphoproteins, indicating that the pattern is dependent on nerve activity, not trophic factors. Such phosphoproteins are possible mediators of neural activity on gene expression.     

An experimental model of mitochondrial myopathy: Germanium-induced myopathy and coenzyme Q10 administration

In skeletal muscles from rats treated with germanium for 23 weeks, there were numerous ragged-red fibers and cytochrome-c oxidase (COX)-deficient fibers. Biochemically, germanium reduced the enzyme activities in the mitochondrial respiratory chain. Rotenone-sensitive NADH–cytochrome-c reductase as well as COX activities were markedly reduced, while succinate–cytochrome-c reductase was less severely, but significantly, affected. The histopathological findings in these muslces were similar to those seen in patients with mitochondrial encephalomyopathy, suggesting that germanium-induced myopathy may be a useful experimental model. Coenzyme Q₁₀ administration appeared to be ineffective in preventing this experimental myopathy. © 1992 John Wiley & Sons, Inc.     

Aerobic exercise and muscle metabolism in patients with mitochondrial myopathy

Exercise therapy improves mitochondrial function in patients with mitochondrial myopathy (MM). We undertook this study to determine the metabolic abnormalities that are improved by exercise therapy. This study identified metabolic pathology using (31)P-magnetic resonance spectroscopy and magnetic resonance imaging (MRI) in a group of patients with MM compared to a control group matched for age, gender, and physical activity. We also observed the effect of exercise therapy for 12 weeks on muscle metabolism and physical function in the MM group. During muscle activity, there was impaired responsiveness of the mitochondria to changes in cytosolic adenosine diphosphate concentration, increased dependence on anaerobic energy pathways, and an adaptive increase in proton efflux in patients with MM. Following exercise therapy, mitochondrial function and muscle mass improved without any change in proton efflux rate. These metabolic findings were accompanied by improvements in functional ability. We conclude that there are significant metabolic differences between patients with MM and a control population, independent of age, gender, and physical activity. Exercise therapy can assist in improving mitochondrial function in MM patients.     

Major growth reduction and minor decrease in mitochondrial enzyme activity in cultured human muscle cells after exposure to zidovudine.

Zidovudine-induced mitochondrial myopathy in AIDS patients reported recently might be due to inhibition of mitochondrial DNA polymerase gamma. We investigated the effect of zidovudine on proliferation, differentiation, activity of mitochondrial- and nuclear-encoded enzymes, and mitochondrial DNA (mtDNA), in cultured human muscle cells. Marked inhibition of cell proliferation was found, even in the presence of low (10 mumol/L) zidovudine concentrations. Enzyme activity of the nuclear-encoded mitochondrial citrate synthase was not affected, and the partially mitochondrial-encoded cytochrome c oxidase was not decreased, except only after exposure to high concentrations (5 mmol/L) zidovudine. No decrease of mtDNA content and no mtDNA deletions were found in zidovudine-exposed muscle cells. We propose that the effect of zidovudine on muscle, seen in zidovudine-treated AIDS patients, results mainly from decrease in proliferation of muscle cells rather than inhibition of mtDNA replication.     

Activities of mitochondrial complexes correlate with nNOS amount in muscle from ALS patients

The pathogenesis of amyotrophic lateral sclerosis (ALS) is poorly understood. Increased levels of free radicals derived from nitric oxide (NO), the product of nitric oxide synthase (NOS), may damage mitochondrial function leading to motor neurone death. Previous studies demonstrated a specific impairment of mitochondrial function in skeletal muscle of ALS patients. In order to verify a pathogenetic relationship between neuronal NOS (nNOS) and mitochondrial function, we studied nNOS expression by Western blot and mitochondrial enzyme activity by spectrophotometric assays in muscle biopsies of 16 sporadic ALS patients and 16 controls subjects. We observed a reduced activity of respiratory chain complexes with mitochondrial encoded subunits and a lower nNOS amount in ALS muscles. There was a direct correlation between levels of nNOS and values of mitochondrial enzymes function. In ALS muscles we found normal levels of manganese superoxide dismutase (SOD2) that is assumed as related to mitochondrial DNA abnormalities. Our data suggest a beneficial role for NO to mitochondrial function and lead to the hypothesis of a common oxidative damage in motor neurones and skeletal muscle in sporadic ALS patients.