Free erythrocyte protoporphyrin level and nerve conduction velocity in end-stage renal disease.

Increased free erythrocyte protoporphyrin concentrations and depressed motor nerve conduction velocities (MNCV) were observed in 45 patients on maintenance haemodialysis. Neither of these findings could be correlated with age, duration or frequency of dialysis, or the degree of uraemia present. A strong negative correlation (r=--0-53; P less than 0-001), however, existed between the free erythrocyte protoporphyrin level and the MNCV, which suggested either (a) a direct effect of iron status on nerve function, or (b) a toxic factor in "uraemia" that depresses both nerve conduction and haemsynthetase activity.     

A novel technique for nonvolitional assessment of quadriceps muscle endurance in humans.

Assessment of quadriceps endurance is of interest to investigators studying human disease. We hypothesized that repetitive magnetic stimulation (rMS) of the intramuscular branches of the femoral nerve could be used to induce and quantify quadriceps endurance. To test this hypothesis, we used a novel stimulating coil to compare the quadriceps endurance properties in eight normal humans and, to confirm that the technique could be used in clinical practice, in eight patients with advanced chronic obstructive pulmonary disease (COPD). To validate the method, we compared in vivo contractile properties of the quadriceps muscle with the fiber-type composition and oxidative enzyme capacity. We used a Magstim Rapid(2) magnetic nerve stimulator with the coil wrapped around the quadriceps. Stimuli were given at 30 Hz, a duty cycle of 0.4 (2 s on, 3 s off), and for 50 trains. Force generation and the surface electromyogram were measured throughout. Quadriceps twitch force, elicited by supramaximal magnetic stimulation of the femoral nerve, was measured before and after the protocol. Quadriceps muscle biopsies were analyzed for oxidative (citrate synthase, CS) and glycolytic (phosphofructokinase, PFK) enzyme activity and myosin heavy chain isoform protein expression. The time for force to fall to 70% of baseline (T(70)) was shorter in the COPD group than the control group: 55.6 +/- 26.0 vs. 121 +/- 38.7 s (P = 0.0014). Considering patients and controls together, positive correlations were observed between T(70) and the proportion of type I fibers (r = 0.68, P = 0.004) and CS-to-PFK ratio (CS/PFK) (r = 0.67, P = 0.005). We conclude that quadriceps endurance assessed using rMS is feasible in clinical studies.     

Myotonia dystrophica: unusual features in a Labrador family.

A large family with myotonia dystrophica has been recognized in an isolated area of Labrador. The complete family tree showed 29 of 108 members to be affected, including an infant with the congenital form of the disease. The propositus presented with epiphora and reduced frequency of blinking, with incomplete closure--features that have not previously been stressed. Ten of the younger affected persons had no lens opacities, although most had systemic muscle signs. Slit-lamp examination was therefore not a valuable method of early detection of the disease in the family. Many of the women affected by myotonia dystrophica had obstetric complications, particularly hydramnios, premature onset of labour, necessity for cesarean section, postpartum hemorrhage and neonatal death. Hydramnios was associated in each instances with perinatal death. The fetus in each case of hydramnios may have had the gene for myotonia dystrophica. Immunoglobulin A concentrations were reduced significantly in 27 affected persons in comparison with 77 unaffected family members. There were no such differences for the other immunoglobulin classes.     

Expression of Muscle-Specific MiRNA 206 in the Progression of Disease in a Murine SMA Model

Spinal muscular atrophy (SMA) is a severe neuromuscular disease, the most common in infancy, and the third one among young people under 18 years. The major pathological landmark of SMA is a selective degeneration of lower motor neurons, resulting in progressive skeletal muscle denervation, atrophy, and paralysis. Recently, it has been shown that specific or general changes in the activity of ribonucleoprotein containing micro RNAs (miRNAs) play a role in the development of SMA. Additionally miRNA-206 has been shown to be required for efficient regeneration of neuromuscular synapses after acute nerve injury in an ALS mouse model. Therefore, we correlated the morphology and the architecture of the neuromuscular junctions (NMJs) of quadriceps, a muscle affected in the early stage of the disease, with the expression levels of miRNA-206 in a mouse model of intermediate SMA (SMAII), one of the most frequently used experimental model. Our results showed a decrease in the percentage of type II fibers, an increase in atrophic muscle fibers and a remarkable accumulation of neurofilament (NF) in the pre-synaptic terminal of the NMJs in the quadriceps of SMAII mice. Furthermore, molecular investigation showed a direct link between miRNA-206-HDAC4-FGFBP1, and in particular, a strong up-regulation of this pathway in the late phase of the disease. We propose that miRNA-206 is activated as survival endogenous mechanism, although not sufficient to rescue the integrity of motor neurons. We speculate that early modulation of miRNA-206 expression might delay SMA neurodegenerative pathway and that miRNA-206 could be an innovative, still relatively unexplored, therapeutic target for SMA.     

Muscle spindle formation and differentiation in regenerating rat muscle grafts

Muscle spindle development and function are dependent upon sensory innervation. During muscle regeneration, both neural and muscular components of spindles degenerate and it is not known whether reinnervation of a regenerating muscle results in reestablishment of proper neuromuscular relationships within spindles or whether sensory neurons may exert an influence upon differentiation of these spindles. Muscle spindle regeneration was studied in bupivacaine-treated grafts of rat extensor digitorum longus (EDL) muscles. Three types of EDL graft were performed in order to manipulate the extent to which regenerating spindles might be reinnervated: (1) grafts reinnervated following severance of their nerve supply (standard grafts); (2) grafts in which intact nerve sheaths appear to facilitate reinnervation (nerveintact grafts); and (3) grafts in which reinnervation was prevented (nonreinnervated grafts). Complete degeneration of muscle fibers occurred in all grafts prior to regeneration. Initial formation of spindles in regenerating EDL grafts is independent of innervation; intrafusal muscle fibers degenerate and regenerate within spindle capsules that remain intact and viable. The extent of spindle differentiation was evaluated in each type of graft using criteria that included nucleation and ATPase activity, both of which have been shown to be regulated by sensory innervation, as well as the number of muscle fibers/spindle and morphology of spindle capsules.While most spindles contained normal numbers of muscle fibers, most of these fibers were morphologically and histochemically abnormal. Alterations of ATPase activity occurred in all spindles, but were least severe in nerve-intact grafts. While fully differentiated nuclear bag and chain fibers were not observed in regenerated spindles, large, vesicular nuclei, similar to those of normal intrafusal fibers, were present in a small number of spindles in nerve-intact grafts. Sensory nerve terminations were observed only in those spindles that also contained the distinctive nuclei. This study suggests that a specific neurotrophic influence is necessary for regeneration of normal intrafusal muscle fibers and that this influence corresponds to the properly timed sensory neuron-muscle interaction which directs muscle spindle embryogenesis. However, the infrequent occurrence of characteristics unique to intrafusal muscle fibers indicates that reinnervation of regenerating muscle grafts by sensory neurons is inadequate and/or faulty.     

Acetylcholinesterase activity in nerve endings of tails of Rana japonica and R. catesbeiana during metamorphosis

Summary In anuran tadpole tails, the myelinated motor nerve fibers branch in the myoseptum to innervate both red and white muscle fibers at, or near, their ends. There are no significant ultrastructural differences between the nerve endings of the two types of muscle fibers.Intense acetylcholinesterase reaction product was observed in synaptic clefts and junctional folds, as well as in transverse tubules. As metamorphosis proceeded, the junctional folds of the nerve endings disappeared, however, acetylcholinesterase reaction product was still observed in the synaptic clefts. As muscle fibers began to degenerate, nerve endings began to separate from them. However, after nerve endings were completely separated from the surfaces, degenerated muscle fibers, synaptic and cored vesicles were still well preserved although no acetylcholinesterase reaction product was found. It seems clear that the mechanism of the muscle degeneration in the tadpole tail during metamorphosis is not the result of the degeneration of its nerve endings.     

An analysis of the interactions between sympathetic nerve fibers and smooth muscle cells in tissue culture

The interactions between sympathetic nerve fibers and smooth muscle cells and fibroblasts from the newborn guinea pig vas deferens were studied in tissue culture with phase contrast microscopy, time-lapse microcinematography, catecholamine fluorescence histochemistry and scanning and transmission electron microscopy. The amount of sympathetic nerve fiber growth, its catecholamine fluorescence reaction and the size of the nerve cell bodies and their nuclei all increased in the presence of vas deferens tissue. Specific growth of nerve fibers to large clumps of vas deferens tissue was seen from distances of up to 2 mm. In contrast, no specific growth from a distance occurred to single cells or small groups of cells. However, random contact with a muscle cell often led to close, extensive, and long-lasting associations. Contact with fibroblasts was always transitory.The rate of sympathetic nerve fiber growth over individual muscle cells was faster than over fibroblasts, which, in turn, was faster than over the collagen-coated surface of the coverslip. Palpation of a muscle cell by a nerve fiber growth cone increased the rate of spontaneous contraction of the muscle cell, the extent of the increase being dependent on the number of nerve fibers involved. Multiple innervation of a smooth muscle cell occurred if nerve fibers reached the cell at about the same time, but not if there was a close association already established. These results are discussed in relation to possible interactions of sympathetic nerve fibers with smooth muscle cells in vivo.     

C 肤对糖尿病大鼠坐骨神经结构和功能的影响

目的：探讨外源性C肽对Ⅰ型糖尿病大鼠坐骨神经结构及功能的影响。方法：选取Wistar大鼠40只,分为正常对照组（NC组）和糖尿病组（Dia组）,糖尿病组链脲佐菌素诱发大鼠成模后,再随机分为三组：糖尿病组（Dia组）、胰岛素治疗组（In组）和C肽治疗组（CP组）8周后,测定各组大鼠运动、感觉神经传导速度,并对病变大鼠的坐骨神经进行病理定量图像分析及超微结构分析,结果：1．In组、CP肽组与DM组相比：大鼠运动、感觉神经传导速度均明显增加（P〈0．01）;2．腓肠神经纤维的数量和总横截面面积也显著增加（P〈0.01）。3．CP组与In组相比运动、感觉神经传导速度也显著增加（P〈0．01）。4．电镜显示：Dia组有髓神经纤维髓鞘发生分离并有无颗粒囊胞状结构聚集现象,In组有髓神经纤维髓鞘分离现象明显减轻但仍有无颗粒囊胞状结构聚集现象．而CP组有髓神经纤维结构完全接近正常组。结论：C肽在改善糖尿病大鼠的神经结构和功能方面明显优于胰岛素．     

Effects of endurance training on activity and expression of AMP-activated protein kinase isoforms in rat muscles

The effects of endurance training on the response of muscle AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) to moderate treadmill exercise were examined. In red quadriceps, there was a large activation of alpha 2-AMPK and inactivation of ACC in response to exercise. This response was greatly reduced after training, probably because of reduced metabolic stress. In white quadriceps, there were no effects of exercise on AMPK or ACC, but alpha 2-activity was higher after training because of increased phosphorylation of Thr(172). In soleus, there were small increases in alpha 2-activity during exercise that were not affected by training. The expression of all seven AMPK subunit isoforms was also examined. The beta 2- and gamma 2-isoforms were most highly expressed in white quadriceps, and gamma 3 was expressed in red quadriceps and soleus. There was a threefold increase in expression of gamma 3 after training in red quadriceps only. Our results suggest that gamma 3 might have a special role in the adaptation to endurance exercise in muscles utilizing oxidative metabolism.     

Inspiratory muscle work in acute hypoxia influences locomotor muscle fatigue and exercise performance of healthy humans

Our aim was to isolate the independent effects of 1) inspiratory muscle work (W(b)) and 2) arterial hypoxemia during heavy-intensity exercise in acute hypoxia on locomotor muscle fatigue. Eight cyclists exercised to exhaustion in hypoxia [inspired O(2) fraction (Fi(O(2))) = 0.15, arterial hemoglobin saturation (Sa(O(2))) = 81 +/- 1%; 8.6 +/- 0.5 min, 273 +/- 6 W; Hypoxia-control (Ctrl)] and at the same work rate and duration in normoxia (Sa(O(2)) = 95 +/- 1%; Normoxia-Ctrl). These trials were repeated, but with a 35-80% reduction in W(b) achieved via proportional assist ventilation (PAV). Quadriceps twitch force was assessed via magnetic femoral nerve stimulation before and 2 min after exercise. The isolated effects of W(b) in hypoxia on quadriceps fatigue, independent of reductions in Sa(O(2)), were revealed by comparing Hypoxia-Ctrl and Hypoxia-PAV at equal levels of Sa(O(2)) (P = 0.10). Immediately after hypoxic exercise potentiated twitch force of the quadriceps (Q(tw,pot)) decreased by 30 +/- 3% below preexercise baseline, and this reduction was attenuated by about one-third after PAV exercise (21 +/- 4%; P = 0.0007). This effect of W(b) on quadriceps fatigue occurred at exercise work rates during which, in normoxia, reducing W(b) had no significant effect on fatigue. The isolated effects of reduced Sa(O(2)) on quadriceps fatigue, independent of changes in W(b), were revealed by comparing Hypoxia-PAV and Normoxia-PAV at equal levels of W(b). Q(tw,pot) decreased by 15 +/- 2% below preexercise baseline after Normoxia-PAV, and this reduction was exacerbated by about one-third after Hypoxia-PAV (-22 +/- 3%; P = 0.034). We conclude that both arterial hypoxemia and W(b) contribute significantly to the rate of development of locomotor muscle fatigue during exercise in acute hypoxia; this occurs at work rates during which, in normoxia, W(b) has no effect on peripheral fatigue.     

Quadriceps and Respiratory Muscle Fatigue Following High-Intensity Cycling in COPD Patients

Exercise intolerance in COPD seems to combine abnormal ventilatory mechanics, impaired O₂ transport and skeletal muscle dysfunction. However their relatie contribution and their influence on symptoms reported by patients remain to be clarified. In order to clarify the complex interaction between ventilatory and neuromuscular exercise limiting factors and symptoms, we evaluated respiratory muscles and quadriceps contractile fatigue, dynamic hyperinflation and symptoms induced by exhaustive high-intensity cycling in COPD patients. Fifteen gold II-III COPD patients (age = 67±6 yr; BMI = 26.6±4.2 kg.m^-2) performed constant-load cycling test at 80% of their peak workload until exhaustion (9.3±2.4 min). Before exercise and at exhaustion, potentiated twitch quadriceps strength (Q_tw), transdiaphragmatic (P_di,tw) and gastric (P_ga,tw) pressures were evoked by femoral nerve, cervical and thoracic magnetic stimulation, respectively. Changes in operational lung volumes during exercise were assessed via repetitive inspiratory capacity (IC) measurements. Dyspnoea and leg discomfort were measured on visual analog scale. At exhaustion, Q_tw (-33±15%, >15% reduction observed in all patients but two) and P_di,tw (-20±15%, >15% reduction in 6 patients) were significantly reduced (P<0.05) but not P_ga,tw (-6±10%, >15% reduction in 3 patients). Percentage reduction in Q_tw correlated with the percentage reduction in P_di,tw (r=0.66; P<0.05). Percentage reductions in P_di,tw and P_ga,tw negatively correlated with the reduction in IC at exhaustion (r=-0.56 and r=-0.62, respectively; P<0.05). Neither dyspnea nor leg discomfort correlated with the amount of muscle fatigue. In conclusion, high-intensity exercise induces quadriceps, diaphragm and less frequently abdominal contractile fatigue in this group of COPD patients. In addition, the rise in end-expiratory lung volume and diaphragm flattening associated with dynamic hyperinflation in COPD might limit the development of abdominal and diaphragm muscle fatigue. This study underlines that both respiratory and quadriceps fatigue should be considered to understand the complex interplay of factors leading to exercise intolerance in COPD patients.     

What controls the outer mitochondrial membrane permeability for ADP: facts for and against the role of oncotic pressure

In our study 10% of bovine serum albumin was added to the physiological incubation medium to mimic the oncotic pressure of the cellular cytoplasm and to test for its effect on the respiration of isolated rat heart mitochondria, saponin- or saponin plus crude collagenase (type IV)-treated heart muscle fibers and saponin-treated rat quadriceps muscle fibers. Pyruvate and malate were used as substrates. We found that albumin slightly decreased the maximal ADP-stimulated respiration rate only for saponin-treated heart muscle fibers. The apparent Km ADP of oxidative phosphorylation increased significantly, by 70-100%, for isolated heart mitochondria, saponin plus collagenase-treated heart muscle fibers and for saponin-treated quadriceps muscle fibers but remained unchanged for saponin-treated heart muscle fibers. The saponin-treated heart muscle fibers were characterized by a very high control apparent Km ADP value (234+/-24 microM ADP) compared with other preparations (14-28 microM ADP). The results suggest that in vivo the oncotic pressure is not the relevant factor causing the low outer mitochondrial membrane permeability for ADP in cardiomyocytes, in contrast to quadriceps muscle cells. It is likely that the outer mitochondrial membrane-bound protein(s) which is supposed to remain in saponin-treated heart muscle fibers is responsible for this property of the membrane.     

Long-term activity in upper- and lower-limb muscles of humans.

Despite limited data on humans, previous studies suggest that there is an association between the duration of daily muscle activity and the proportion of type I muscle fibers. We quantified the activity of limb muscles in healthy men and women during normal use and compared these measurements with published reports on fiber-type proportions. Seven men (age range = 21-28 yr) and seven women (age range = 18-26 yr) participated in two 10-h recording sessions. Electromyogram (EMG) activity of four muscles in nondominant upper (first dorsal interosseus and biceps brachii) and lower limbs (vastus medialis and vastus lateralis) was recorded with surface electrodes. Hand and arm muscles were active for 18% of the recording time, whereas leg muscles were active for only 10% of the recording time. On average, upper-limb muscles were activated 67% more often than lower-limb muscles. When lower-limb muscles were activated, however, the mean amplitude of each burst was greater in leg muscles [18 and 17% maximum voluntary contraction (MVC)] compared with hand (8% MVC) and arm (6% MVC) muscles. Temporal association in activity between pairs of muscles was high for the two lower-limb muscles (r2 = 0.7) and relatively weak for the two upper-limb muscles (r2 = 0.09). Long-term muscle activity was only different between men and women for the biceps brachii muscle. We found no relation between duration of muscle activity in 10-h recordings and the reported values of type I fibers in men and women.     

Ultrastructural changes in the red muscle fibers of the quadriceps muscle of the rat femur during increased motor activity

Experiments on rats were made to study the effect of physical exercise running in a treadbahn on ultrastructure of skeletal muscle fibers. The biphasic mechanism of muscle contractile activity was shown. The processes of destruction occurring in red muscle fibers of the intensely working quadriceps femoris were manifested by enlargement of T system and sarcoplasmic reticulum elements, mitochondrial matrix swelling, cryst destruction, vacuolar degeneration of part of the mitochondria, and destruction of individual myofibrils. In addition to destructive changes, the muscle exhibited the recovery processes--physiological regeneration. Those processes included large accumulations of the mitochondria beneath the plasma membrane of muscle fibers, the presence of small mitochondria, division of the mitochondria, transformation of myosatellitocytes to myoblasts, the presence of centrioles in endotheliocytes, and so forth.     

Treatment of diabetic polyneuropathy with the antioxidant thioctic acid (alpha-lipoic acid): a two year multicenter randomized double-blind placebo-controlled trial (ALADIN II). Alpha Lipoic Acid in Diabetic Neuropathy.

Short-term trials with the antioxidant thioctic acid (TA) appear to improve neuropathic symptoms in diabetic patients, but the long-term response remains to be established. Therefore, Type 1 and Type 2 diabetic patients with symptomatic polyneuropathy were randomly assigned to three treatment regimens: (1) 2 x 600(mg of TA (TA 1200), (2) 600)mg of TA plus placebo (PLA) (TA 600) or (3) placebo and placebo (PLA). A trometamol salt solution of TA of 1200 or 600 mg or PLA was intravenously administered once daily for five consecutive days before enrolling the patients in the oral treatment phase. The study was prospective, PLA-controlled, randomized, double-blind and conducted for two years. Severity of diabetic neuropathy was assessed by the Neuropathy Disability Score (NDS) and electrophysiological attributes of the sural (sensory nerve conduction velocity (SNCV), sensory nerve action potential (SNAP)) and the tibial (motor nerve conduction velocity (MNCV), motor nerve distal latency (MNDL)) nerve. Statistical analysis was performed after independent reviewers excluded all patients with highly variable data allowing a final analysis of 65 patients (TA 1200: n = 18, TA 600: n = 27; PLA: n = 20). At baseline no significant differences were noted between the groups regarding the demographic variables and peripheral nerve function parameters for these 65 patients. Statistically significant changes after 24 months between TA and PLA were observed (mean +/- SD) for sural SNCV: +3.8 +/- 4.2 m/s in TA 1200, +3.0+/-3.0m/s in TA 600, -0.1+/-4.8m/s in PLA (p < 0.05 for TA 1200 and TA 600 vs. PLA); sural SNAP: +0.6+/-2.5 microV in TA 1200, +0.3+/-1.4 microV in TA 600, -0.7 +/- 1.5 microV in PLA (p = 0.076 for TA 1200 vs. PLA and p < 0.05 for TA 600 vs. PLA), and in tibial MNCV: +/- 1.2 +/- 3.8 m/s in TA 1200, -0.3 +/- 5.2 m/s in TA 600, 1.5 +/- 2.9 m/s in PLA (p < 0.05 for TA 1200 vs. PLA). No significant differences between the groups after 24 months were noted regarding the tibial MNDL and the NDS. We conclude that in a subgroup of patients after exclusion of patients with excessive test variability throughout the trial, TA appeared to have a beneficial effect on several attributes of nerve conduction.     

Multiple Sclerosis Affects Skeletal Muscle Characteristics

Background

The impact of multiple sclerosis (MS) on skeletal muscle characteristics, such as muscle fiber cross sectional area (CSA), fiber type proportion, muscle strength and whole muscle mass, remains conflicting.

Methods

In this cross sectional study, body composition and muscle strength of the quadriceps were assessed in 34 MS (EDSS: 2.5±0.19) patients and 18 matched healthy controls (HC). Hereafter a muscle biopsy (m.vastus lateralis) was taken.

Results

Compared to HC, mean muscle fiber CSA of all fibers, as well as CSA of type I, II and IIa fibers were smaller and muscle strength of the quadriceps was lower in MS patients. Whole body composition was comparable between groups. However, compared to HC, the biopsied leg tended to have a higher fat percentage (p = 0.1) and a lower lean mass (p = 0.06) in MS patients.

Conclusion

MS seems to negatively influence skeletal muscle fiber CSA, muscle strength and muscle mass of the lower limbs of mildly affected MS patients. This emphasises the need for rehabilitation programs focusing on muscle preservation of the lower limb.

Trial Registration

ClinicalTrials.gov NCT01845896     

Motor innervation of the pharynx levator muscle of the snail, Helix pomatia: physiological and histological properties

(1) Motor innervation of the pharynx levator muscle of Helix pomatia was investigated with intracellular recording and axonal iontophoresis of cobalt chloride. (2) Muscle fibers respond to direct electrical stimulation of the muscle with active graded responses or non-overshooting spike potentials. (3) Each fiber is innervated via the external and internal lip nerves by several (mostly 3) excitatory nerve fibers each. Two types of EPSPs can be distinguished according to amplitude, duration, and facilitation. (4) Axonal CoCl2-staining via an external lip nerve branch revealed many nerve fibers entering the muscle and branching there into a rich network of blebbed fibers of various diameters.     

Morphometric comparison between human and rat abducens and oculomotor nerves

A morphologic and morphometric comparison between normal human and rat extraocular muscle nerves was performed using a computer-assisted method to obtain scatter diagrams of relative sheath thickness (g ratio = quotient axon diameter/fiber diameter). Human and rat extraocular muscle nerves (nervus abducens and ramus medialis n. oculomotorii) were excised immediately before the nerve branching at the entering point into the muscle. There was no difference in the absolute number of myelinated fibers between the oculomotor and abducens nerves in both species. The distribution of myelinated fibers was classified according to their g ratios into a two-stage density cluster analysis. Two main populations of nerve fibers for human oculomotor and rat oculomotor and abducens nerves and three main populations for human abducens nerve were differentiated morphometrically and mathematically, differing in their relative sheath thicknesses. There are distinct differences between scatter diagrams of human and rat extraocular muscle nerves, in correlation with the basically different oculomotor functions of these two species. The morphometric differences between human and rat extraocular muscle nerves suggest a difference in the myelination process and the presence of functionally different nerve fibers, strongly indicated by the populations and subpopulations of myelinating nerve fibers peculiar to extraocular muscle. The existence of more than two different types of myelinated fibers in the human nerves implies that the traditional classification based on fiber caliber must be reviewed and a comparison of different classes of nerve and muscle fibers should be performed.