Differential calcineurin signalling activity and regeneration efficacy in diaphragm and limb muscles of dystrophic mdx mice

Calcineurin activity is essential for successful skeletal muscle regeneration in young mdx mice and in wild type mice following myotoxic injury and cryodamage. In mature myofibres of adult mdx mice, calcineurin stimulation can ameliorate the dystrophic pathology. The aim of this study was to test the hypothesis that the more severe dystrophic pathology of the diaphragm compared with hindlimb muscles of mdx mice could be attributed to aberrant calcineurin signalling and that due to ongoing regeneration calcineurin activity would be greater in muscles of adult mdx than wild type mice. Differences in markers of regeneration between tibialis anterior and diaphragm muscles were also characterised, to determine whether there was an association between regeneration efficacy and calcineurin activity in dystrophic muscles. In diaphragm muscles of adult mdx mice, the proportion of centrally nucleated fibres and developmental myosin heavy chain protein expression was lower and myogenin protein expression was higher than in tibialis anterior muscles. Calcineurin and activated NFATc1 protein content and calcineurin phosphatase activity were higher in muscles from mdx than wild type mice and calcineurin activation was greater in diaphragm than tibialis anterior muscles of mdx mice. Thus, despite greater calcineurin activity in diaphragm compared to hindlimb muscles, regeneration events downstream of myoblast differentiation and mediated by the injured myofibre were severely compromised.

Examination of EMG normalisation methods for the study of the posterior and posterolateral neck muscles in healthy controls

The purpose of this study was to examine the reliability of normalisation methods used in the study of the posterior and posterolateral neck muscles in a group of healthy controls. Six asymptomatic male subjects performed a total of 12 maximum voluntary isometric contractions (MVIC) and 60%-submaximal isometric contractions (60%-MVIC) against the torque arm of an isokinetic dynamometer whilst surface and intramuscular electromyography (EMG) was recorded unilaterally from representative posterior and posterolateral locations. Reliability was calculated using intra-class correlation coefficient (ICC), relative standard error of measurement (%SEM) and relative coefficient of variation (%CV). Maximal torque output was found to be highly reliable in the directions of extension and right lateral bending when the first of three MVIC contractions was excluded. When averaged across contraction direction, high reliability was found for both surface (MVIC: ICC = 0.986, %SEM = 7.5, %CV = 9.2; 60%-MVIC: ICC = 0.975, %SEM = 10, %CV = 13.7) and intramuscular (MVIC: ICC = 0.910, %SEM = 20, %CV = 19.1; 60%-MVIC: ICC = 0.952, %SEM = 16.5, %CV = 13.5) electrodes. Intramuscular electrodes displayed the least reliability in right lateral bending. The use of visual feedback markedly increased the reliability of 60%-MVIC contractions.

Reliability of normalisation methods for EMG analysis of neck muscles

Acceptable reliability of normalisation contractions in electromyography (EMG) is paramount for testing conducted over a number of days or if normal laboratory strength testing equipment is unavailable. This study examined the reliability of maximal voluntary isometric contractions (MVIC) and sub-maximal (60%) isometric contractions for use in neck muscle EMG studies. Surface EMG was recorded bilaterally from eight sites around the neck at C4/5 level from five healthy male subjects. Subjects performed MVIC and sub-maximal normalisation contractions using an isokinetic dynamometer (ID) and a portable cable dynamometer with attached strain gauge (PCD) in addition to a MVIC against a manual resistance (MR). Subjects were tested in flexion, extension, left and right lateral bending and were retested by the same tester within a two-week period. Intra class correlation co-efficients (ICC) were calculated for each testing method and contraction direction and a mean ICC was calculated across all contraction directions. All normalisation methods produced excellent within-day reliability (mean ICC >0.80) but only the MVICs using the ID and PCD had acceptable reliability when assessed between-days. This study confirmed the validity of using MVICs elicited using the ID and PCD as reliable reference contractions for the normalisation of neck EMG.

Endurance training adaptations modulate the redox–force relationship of rat isolated slow-twitch skeletal muscles

1. Studies have shown that, in isolated skeletal muscles, maximum isometric force production (Po) is dependent on muscle redox state. Endurance training increases the antioxidant capacity of skeletal muscles, a factor that could impact on the force-producing capacity following exogenous exposure to an oxidant. We tested the hypothesis that 12 weeks treadmill training would increase anti-oxidant capacity in rat skeletal muscles and alter their response to exogenous oxidant exposure.

2. At the conclusion of the 12 week endurance-training programme, soleus (slow-twitch) muscles from trained rats had greater citrate synthase (CS) and catalase (CAT) activity compared with soleus muscles from untrained rats (P < 0.05).
In contrast, CAT activity of extensor digitorum longus (EDL; fast-twitch) muscles from trained rats was not different to EDL muscles of untrained rats. The CS activity was lower in EDL muscles from trained compared with untrained rats (P < 0.05).

3. Equilibration with exogenous hydrogen peroxide (H2O2, 5 mmol/L) increased the Po of soleus muscles from untrained rats for the duration of treatment (30 min), whereas the Po of EDL muscles was affected biphasically, with a small increase initially (after 5 min), followed by a more marked decrease in Po (after 30 min). The H2O2-induced increase in Po of soleus muscles from trained rats was less than that in untrained rats (P < 0.05), but no differences were observed in the Po of EDL muscles following training.

4. The results indicate that 12 weeks endurance running training conferred adaptations in soleus but not EDL muscles. These adaptations were associated with an attenuation of the oxidant-induced increase in Po of soleus muscles from trained compared with untrained rats. We conclude that endurance training-adapted soleus muscles have a slightly altered redox - force relationship.

Oxygen dependence of metabolism and cellular adaptation in vertebrate muscles: a review

The key roles the cardiovascular system play in the complex distribution of blood, and consequently oxygen, have been extensively studied in vertebrates. Numerous studies have also revealed the complex and varied ways in which tissues cope with compromised oxygen supply. The links between these two processes are the subject of much current research. This article aims to review how blood supply influences tissue oxygenation and affects metabolism, and how this might have played a role in the evolution of the complex muscle arrangements which characterise vertebrates. Muscle tissue is the greatest proportion of body mass in most vertebrates and undergoes dramatic alterations in metabolism and associated oxygen flux. Special attention is given to the myotome of fishes, in which the partitioning of the fibre types contrasts with the mosaic arrangement of tetrapods. This gives us the opportunity to study pure whole vascularised muscle blocks, rather than single fibres, and further explore the interrelationship between oxygen supply and tissue energetics.

Oxygen-dependence of metabolic rate in the muscles of craniates

We present evidence that oxygen consumption (V_O2 ) is oxygen partial pressure (P_O2) dependent in striated muscles and P_O2 -independent in the vasculature in representatives of three craniate taxa: two teleost fish, a hagfish and a rat. Blood vessel V_O2 displayed varying degrees of independence in a P_O2 range of 15–95 mmHg, while V_O2 by striated muscle tissue slices from all species related linearly to P_O2 between 0 and 125 mmHg, despite V_O2 rates varying greatly between species and muscle type. In salmon red muscle, lactate concentrations fell in slices incubated at a P_O2 of either 30 or 100 mmHg, suggesting aerobic rather than anaerobic metabolism. Consistent with this finding, potential energy, a proxy of ATP turnover, was P_O2 -dependent. Our data suggest that the reduction in V_O2 with falling P_O2 results in a decrease in ATP demand, suggesting that the hypoxic signal is sensed and cellular changes effected. Viability and diffusion limitation of the preparations were investigated using salmon cardiac and skeletal muscles. Following the initial P_O2 depletion, reoxygenation of the Ringer bathing salmon cardiac muscle resulted in V_O2^s that was unchanged from the first run. V_O2 increased in all muscles uncoupled with p-trifluoromethoxylphenyl-hydrazone (FCCP) and 2,4-dinitrophenol (DNP). Mitochondrial succinate dehydrogenase activity, quantified by reduction of 3-(4,5-dimethylthiazol)-2,5-diphenyl-2H-tetrazolium bromide (MTT) to formazan, was constant over the course of the experiment. These three findings indicate that the tissues remained viable over time and ruled out diffusion-limitation as a constraint on V_O2.

Artificial Muscles Based on Polypyrrole/Carbon Nanotube Laminates

A novel method for the periodical assembly of laminates of forest-drawn carbon nanotube (CNT) sheets and polypyrrole (PPy) is described. The method produces composite films in which the volume fraction and orientation of CNTs can be controlled. Actuator stroke and strength is increased and work capacity per cycle doubled when nanotube orientation is perpendicular to the actuation direction. Most importantly, these PPy/CNT laminates have dramatically decreased creep during actuation, which has been a major barrier for the application of PPy actuators.

Alterations in Notch signalling in skeletal muscles from mdx and dko dystrophic mice and patients with Duchenne muscular dystrophy.

New Findings What is the central question of this study? The Notch signalling pathway plays an important role in muscle regeneration, and activation of the pathway has been shown to enhance muscle regeneration in aged mice. It is unknown whether Notch activation will have a similarly beneficial effect on muscle regeneration in the context of Duchenne muscular dystrophy (DMD). What is the main finding and its importance? Although expression of Notch signalling components is altered in both mouse models of DMD and in human DMD patients, activation of the Notch signalling pathway does not confer any functional benefit on muscles from dystrophic mice, suggesting that other signalling pathways may be more fruitful targets for manipulation in treating DMD. Abstract In Duchenne muscular dystrophy (DMD), muscle damage and impaired regeneration lead to progressive muscle wasting, weakness and premature death. The Notch signalling pathway represents a central regulator of gene expression and is critical for cellular proliferation, differentiation and apoptotic signalling during all stages of embryonic muscle development. Notch activation improves muscle regeneration in aged mice, but its potential to restore regeneration and function in muscular dystrophy is unknown. We performed a comprehensive examination of several genes involved in Notch signalling in muscles from dystrophin-deficient mdx and dko (utrophin- and dystrophin-null) mice and DMD patients. A reduction of Notch1 and Hes1 mRNA in tibialis anterior muscles of dko mice and quadriceps muscles of DMD patients and a reduction of Hes1 mRNA in the diaphragm of the mdx mice were observed, with other targets being inconsistent across species. Activation and inhibition of Notch signalling, followed by measures of muscle regeneration and function, were performed in the mouse models of DMD. Notch activation had no effect on functional regeneration in C57BL/10, mdx or dko mice. Notch inhibition significantly depressed the frequency-force relationship in regenerating muscles of C57BL/10 and mdx mice after injury, indicating reduced force at each stimulation frequency, but enhanced the frequency-force relationship in muscles from dko mice. We conclude that while Notch inhibition produces slight functional defects in dystrophic muscle, Notch activation does not significantly improve muscle regeneration in murine models of muscular dystrophy. Furthermore, the inconsistent expression of Notch targets between murine models and DMD patients suggests caution when making interspecies comparisons.

Sociocultural influences on strategies to lose weight, gain weight, and increase muscles among ten cultural groups

This study determined how sociocultural messages to change one's body are perceived by adolescents from different cultural groups. In total, 4904 adolescents, including Australian, Chilean, Chinese, Indo-Fijian, Indigenous Fijian, Greek, Malaysian, Chinese Malaysian, Tongans in New Zealand, and Tongans in Tonga, were surveyed about messages from family, peers, and the media to lose weight, gain weight, and increase muscles. Groups were best differentiated by family pressure to gain weight. Girls were more likely to receive the messages from multiple sociocultural sources whereas boys were more likely to receive the messages from the family. Some participants in a cultural group indicated higher, and others lower, levels of these sociocultural messages. These findings highlight the differences in sociocultural messages across cultural groups, but also that adolescents receive contrasting messages within a cultural group. These results demonstrate the difficulty in representing a particular message as being characteristic of each cultural group.