Deubiquitinating enzymes in skeletal muscle atrophy—An essential role for USP19

The ubiquitin proteasome system is well recognized to be involved in mediating muscle atrophy in response to diverse catabolic conditions. To date, almost all of the genes that have been implicated are ubiquitin ligases. Although ubiquitination is modulated also by deubiquitinating enzymes, the roles of these enzymes in muscle wasting remains largely unexplored. In this article, the potential roles of deubiquitinating enzymes in regulating muscle size are discussed. This is followed by a review of the roles described for USP19, the deubiquitinating enzyme that has been most studied in muscle wasting. This enzyme is upregulated in muscle in many catabolic conditions and its inactivation leads to protection from muscle loss induced by stimuli that are common in many illnesses causing cachexia. It can regulate both protein synthesis and protein degradation as well as myogenesis, thereby modulating the key processes that control muscle mass. Roles for other deubiquitinating enzymes remain possible and to be explored.     

The recruitment order of scapular muscles depends on the characteristics of the postural task

Previous studies show that the scapular muscle recruitment order could possibly change according to the characteristics of the postural task. We aimed to compare the activation latencies of serratus anterior (SA), upper, middle, and lower trapezius (UT, MT and LT, respectively) between an unpredictable perturbation (sudden arm destabilization) and a predictable task (voluntary arm raise) and, to determine the differences in the muscle recruitment order in each task. The electromyographic signals of 23 participants were recorded while the tasks were performed. All scapular muscles showed earlier onset latency in the voluntary arm raise than in the sudden arm destabilization. No significant differences were observed in the muscle recruitment order for the sudden arm destabilization (p > 0.05). Conversely, for voluntary arm raise the MT, LT SA and anterior deltoid (AD) were activated significantly earlier than the UT (p < 0.001). Scapular muscles present a specific recruitment order during a predictable task: SA was activated prior to the AD and the UT after the AD, in a recruitment order of SA, AD, UT, MT, and LT. While in an unpredictable motor task, all muscles were activated after the destabilization without a specific recruitment order, but rather a simultaneous activation.     

Effects of whole body vibration on muscle contractile properties in exercise induced muscle damaged females

Determining muscle contractile properties following exercise is critical in understanding neuromuscular function. Following high intensity training, individuals often experience exercise induced muscle damage (EIMD). The purpose of this investigation was to determine the effect of whole-body vibration (WBV) on muscle contractile properties following EIMD. Twenty-seven females volunteered for 7 sessions and were randomly assigned to a treatment or control group. Muscle contractile properties were assessed via voluntary torque (VT), peak twitch torque (TT), time to reach peak torque, half relaxation time of twitch torque, percent activation (%ACT), rate of rise (RR), rate of decline (RD), mean and peak electromyography during maximum voluntary isometric contraction. Two testing sets were collected each day, consisting of pre measures followed by WBV or control and post measures. A mixed factor analysis of variance was conducted for each variable. %ACT measures found baseline being less than day 1 in both measures in the control group. TT was found to be greater in the control group compared to WBV group. TT and VT baseline measures were greater than all other time points. RR showed control group had higher values than WBV group. These results indicate that WBV following EIMD had some positive effects on muscle contractile properties.     

Intracellular accumulation of trehalose and glycogen in an extreme oligotroph,Rhodococcus erythropolis N9T-4

An extreme oligotroph, Rhodococcus erythropolis N9T-4, showed intracellular accumulation of trehalose and glycogen under oligotrophic conditions. No trehalose accumulation was observed in cells grown on the rich medium. Deletion of the polyphosphate kinase genes enhanced the trehalose accumulation and decreases the intracellular glycogen contents, suggesting an oligotrophic relationship between among the metabolic pathways of trehalose, glycogen, and inorganic polyphosphate biosyntheses.     

髁突-翼外肌解剖复位与游离复位治疗髁状突骨折的疗效比较

目的:比较髁突-翼外肌解剖复位与游离复位治疗髁状突骨折的疗效,促进髁突形态恢复。方法:收治的80例单侧髁状突骨折患者随机分为两组,每组40例,A组行髁突-翼外肌解剖复位术,B组行髁状突游离复位术,术后3个月、6个月观察髁突形态及下颌骨运动功能变化。结果:A组治愈率为90%,高于B组的70.00%(P0.05);术后3个月A组髁状突吸收、张口受限、开口偏斜、咬合关系紊乱、关节弹响发生率分别为12.50%、15.00%、15.00%、7.50%、12.50%,均低于B组的32.50%、35.00%、37.50%、25.00%、35.00%(P0.05);术后6个月A组张口受限、关节弹响发生率为5.00%、2.50%,均低于B组的20.00%、20.00%(P0.05);两组术后并发症发生率比较差异无统计学意义(P0.05)。结论:髁突-翼外肌解剖复位术保留髁状突骨折患者骨折断端血运,髁突形态及下颌骨运动能力恢复良好,疗效优于髁状突游离复位术。     

Expression of cardiac function genes in adult stem cells is increased by treatment with nitric oxide agents

Mesenchymal stem cells (MSCs) have received special attention for cardiomyoplasty because several studies have shown that they differentiate into cardiomyocytes both in vitro and in vivo. Nitric oxide (NO) is a free radical signaling molecule that regulates several differentiation processes including cardiomyogenesis. Here, we report an investigation of the effects of two NO agents (SNAP and DEA/NO), able to activate both cGMP-dependent and -independent pathways, on the cardiomyogenic potential of bone marrow-derived mesenchymal stem cells (BM-MSCs) and adipose tissue-derived stem cells (ADSCs). The cells were isolated, cultured and treated with NO agents. Cardiac- and muscle-specific gene expression was analyzed by indirect immunofluorescence, flow cytometry, RT-PCR and real-time PCR. We found that untreated (control) ADSCs and BM-MSCs expressed some muscle markers and NO-derived intermediates induce an increased expression of some cardiac function genes in BM-MSCs and ADSCs. Moreover, NO agents considerably increased the pro-angiogenic potential mostly of BM-MSCs as determined by VEGF mRNA levels.     

Seasonal changes in the activation of crossbridge motions of isolated thick filament from Limulus striated muscle

Summary In dynamic light scattering, measurements of the intensity-intensity time correlation function from a suspension of rod-like particles of length L could reveal dynamical information related to translational and internal motions of those particles. For a suspension of thick filaments isolated from the myosin-regulated, striated muscles of Limulus at KL>1 (where K is the scattering vector), the average characteristic linewidth ( ) increased with the addition of Ca²⁺ or with the depletion of ATP. The increase in the with the addition of Ca²⁺ could be due to the presence of energy-requiring, high-frequency motions of the crossbridges activated by Ca²⁺. The increase in which occurred with the depletion of ATP was assumed to be mainly due to the thermal motions of the crossbridges after they had moved radially away from the filament backbone. The percentage increase in following the addition of Ca²⁺ was found to be seasonal, i.e., values of obtained from thick filaments isolated between the middle of June and the middle of September were smaller than those obtained during the rest of the year. The effect of temperature on the percentage increase in was also different. The increase showed a maximum at about 35°C during the summer and at about 25°C at other times. However, the percentage increase in developed under ATP-depleted conditions showed no temperature-related maximum. The number of bound Ca²⁺ per myosin molecule was 1 during the summer and 2 at other times.Abbreviations DLS dynamic light scattering - L length - K scattering vector - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis - average characteristic line width Deceased     

The Superfast Human Extraocular Myosin Is Kinetically Distinct from the Fast Skeletal IIa,IIb, and IId Isoforms

Humans express five distinct myosin isoforms in the sarcomeres of adult striated muscle (fast IIa, IId, the slow/cardiac isoform I/β, the cardiac specific isoform α, and the specialized extraocular muscle isoform). An additional isoform, IIb, is present in the genome but is not normally expressed in healthy human muscles. Muscle fibers expressing each isoform have distinct characteristics including shortening velocity. Defining the properties of the isoforms in detail has been limited by the availability of pure samples of the individual proteins. Here we study purified recombinant human myosin motor domains expressed in mouse C₂C₁₂ muscle cells. The results of kinetic analysis show that among the closely related adult skeletal isoforms, the affinity of ADP for actin·myosin (K_AD) is the characteristic that most readily distinguishes the isoforms. The three fast muscle myosins have K_AD values of 118, 80, and 55 μm for IId, IIa, and IIb, respectively, which follows the speed in motility assays from fastest to slowest. Extraocular muscle is unusually fast with a far weaker K_AD = 352 μm. Sequence comparisons and homology modeling of the structures identify a few key areas of sequence that may define the differences between the isoforms, including a region of the upper 50-kDa domain important in signaling between the nucleotide pocket and the actin-binding site.     

Isolation of Pulmonary Artery Smooth Muscle Cells from Neonatal Mice

Pulmonary hypertension is a significant cause of morbidity and mortality in infants. Historically, there has been significant study of the signaling pathways involved in vascular smooth muscle contraction in PASMC from fetal sheep. While sheep make an excellent model of term pulmonary hypertension, they are very expensive and lack the advantage of genetic manipulation found in mice. Conversely, the inability to isolate PASMC from mice was a significant limitation of that system. Here we described the isolation of primary cultures of mouse PASMC from P7, P14, and P21 mice using a variation of the previously described technique of Marshall et al.²⁶ that was previously used to isolate rat PASMC. These murine PASMC represent a novel tool for the study of signaling pathways in the neonatal period. Briefly, a slurry of 0.5% (w/v) agarose + 0.5% iron particles in M199 media is infused into the pulmonary vascular bed via the right ventricle (RV). The iron particles are 0.2 μM in diameter and cannot pass through the pulmonary capillary bed. Thus, the iron lodges in the small pulmonary arteries (PA). The lungs are inflated with agarose, removed and dissociated. The iron-containing vessels are pulled down with a magnet. After collagenase (80 U/ml) treatment and further dissociation, the vessels are put into a tissue culture dish in M199 media containing 20% fetal bovine serum (FBS), and antibiotics (M199 complete media) to allow cell migration onto the culture dish. This initial plate of cells is a 50-50 mixture of fibroblasts and PASMC. Thus, the pull down procedure is repeated multiple times to achieve a more pure PASMC population and remove any residual iron. Smooth muscle cell identity is confirmed by immunostaining for smooth muscle myosin and desmin.     

Splicing of the Muscle-Specific Plasma Membrane Ca2+-ATPase Isoform PMCA1c Is Associated with Cell Fusion in C2 Myocytes

Abstract: The regulation of intracellular calcium is essential for proper muscle function. Muscle cells have several mechanisms for dealing with the rapid and large changes in cytosolic calcium level that occur during contraction. Among these is the plasma membrane Ca²⁺-ATPase (PMCA), which pumps calcium from the cytosol to the extracellular space. We have previously shown that in human fetal muscle the PMCA1 isoforms present are PMCA1a-d, with PMCA1b and c predominating. Alternative splicing of mRNAs encoding proteins involved in muscle contraction is common in developing muscle. Therefore, we examined the expression of muscle-specific PMCA mRNAs in pre- and postfusion mouse C2 myoblasts. The housekeeping form of the Ca²⁺-ATPase, PMCA1b, was found at all times and under all conditions. However, the other predominating isoform found in muscle, PMCA1c, was expressed on myotube formation. Simple cell-cell contact was not sufficient to induce PMCA1c expression, as cells plated at confluence but harvested before myotube formation did not express PMCA1c. The induction of this muscle-specific Ca²⁺-ATPase at myotube formation suggests that it may play an important role in muscle function.