Autophagy is a ubiquitous process through which damaged cytoplasmic structures are recycled and degraded within cells. Aging can affect autophagy regulation in different steps leading to the accumulation of damaged organelles and proteins, which can contribute to cell dysfunction and death. Motor neuron (MN) loss and sarcopenia are prominent features of neuromuscular aging. Previous studies on phrenic MNs showed increased levels of the autophagy proteins LC3 and p62 in 24 month compared to 6 month old mice, consistent with the onset of diaphragm muscle sarcopenia. In the present study, we hypothesized that aging leads to increased expression of the autophagy markers LC3 and p62 in single lumbar MNs. Expression of LC3 and p62 in lumbar MNs (spinal levels L1-L6) was assessed using immunofluorescence and confocal imaging of male and female mice at 6, 18 and 24 months of age, reflecting 100 %, 90 % and 75 % survival, respectively. A mixed linear model with animal as a random effect was used to compare relative LC3 and p62 expression in choline acetyl transferase-positive MNs across age groups. Expression of LC3 and p62 decreased in the white matter of the lumbar spinal cord with aging, with ~29 % decrease in LC3 and ~ 7 % decrease in p62 expression at 24 months of age compared to 6 months of age. There was no change in LC3 or p62 expression in the gray matter with age. LC3 expression in MNs relative to white matter increased significantly with age, with 150 % increase at 24 months of age compared to 6 months of age. Similarly, p62 expression in MNs relative to white matter increased significantly with age, with ~14 % increase at 24 months of age compared to 6 months of age. No effect of sex or MN pool was observed in LC3 and p62 expression in MNs. Overall, these data suggest autophagy impairment during elongation (increased LC3) and degradation (increased p62) phases with aging in lumbar MNs.

Muscle injuries make up the majority of injuries in popular sports. The causes for the development are multifactorial and can be divided into functional disorders and a lack of knowledge regarding training control. Insufficient warm-up and overtraining both increase the danger of the occurrence of the injury. Knowledge of the different types of muscle injuries and their incorporation into an exact classification enable the establishment of a correct treatment plan and can have a positive influence on the healing process. In addition to acute treatment the knowledge and application of preventive programs are necessary as these can significantly reduce the prevalence of muscle injuries in sports.
UNASSIGNED: Muskelverletzungen nehmen einen großen Teil der Verletzungen im Breitensport ein. Die Ursachen für die Entstehung sind multifaktoriell und lassen sich in funktionelle Störungen und falsche Trainingssteuerung gliedern. Sowohl ungenügendes Aufwärmen als auch ein Übertraining erhöhen die Gefahr in der Entstehung der Verletzung. Die Kenntnis der unterschiedlichen Verletzungsarten der Muskelverletzungen und die Eingliederung in eine exakte Klassifikation ermöglichen das Aufstellen eines korrekten Therapieplans. Neben der Akuttherapie sind die Kenntnis und die Anwendung von Präventionsprogrammen notwendig, da diese die Prävalenz von Muskelverletzungen im Sport reduzieren können.

Immobilization-induced Neuromuscular Dysfunction (NMD) increases morbidity and mortality of patients in Intensive Care Units. However, the underlying mechanism of NMD remain poorly elucidated which limited the development of therapeutic method for NMD. Here we developed an immobilization rat model and tested the hypothesis that decreased expression of NRG-1, abnormal expression and distribution of nicotinic acetylcholine receptors (nAChRs) in skeletal muscle caused by immobilization can lead to NMD. To investigate the role of NRG-1/ErbB pathway on immobilization-induced NMD, exogenous recombinant human neuregulin-1 (rhNRG-1) was used to increase the expression of NRG-1 in skeletal muscle during immobilization. It was observed rhNRG-1 significantly alleviated the muscle loss and enhanced the expression of ε-nAChR, while diminished the expression of γ- and α7-nAChR and NMD. Interestingly, ErbB inhibitor PD158780 blocked the protective effects of rhNRG-1. Collectively, the results of present study suggested that rhNRG-1 attenuated immobilization-induced muscle loss and NMD, suppressed γ- and α7-nAChR production, enhanced ε-nAChR synthesis via activating NRG-1/ErbB pathway. Taken together, our findings provide novel insights into NMD contribution, suggesting that the rhNRG-1 is a promising therapy to protect against immobilization-induced myopathy.

Simple reaction times (SRTs), measured in milliseconds (msec), are equal to the sum of subject-dependent latencies that occur during cognitive processing and neuromuscular responses to a preprogrammed stimulus presented to a subject. SRTs have the advantage of being a relatively pure, neurologically driven motor/sensory task that provides a clinician with a generalized assessment of functional deficits. SRTs are easily obtained, and studies have reported that the average number of finger-taps during a 10-s interval can be utilized to distinguish between patients with mild traumatic brain injury (TBI) and healthy controls.
A stimulus/response SRT protocol, utilizing an iPad-based tool, was utilized to see if differentiation and quantification of individual components of cognitive latency (CL) and neuromuscular latency (NL) from a subject\'s total SRT could be accomplished. The study hypothesis was that cognitive latencies, related to a specific cognitive challenge, would remain constant even as latencies due to neuromuscular challenges changed.
Two categories of SRT tests were utilized. The first was a simple finger tapping test (FTT), without any cognitive involvement, that was designed to quantify pure NL. The second test was a choice test (CT) that was characterized by the addition of a cognitive task to the FTT. The objective of the FTT was for the subject to simply tap a single target positioned on the iPad screen as fast as possible over an interval of 10 s. Measurement of the SRT began when a target was displayed and ended when the target was tapped. Two levels of challenge for the FTT and CT tests were presented to the subjects: a small random displacement (SD) of the target of up to 6 mm, and a large random displacement (LD) of the target of up to 24 mm. It was expected that the magnitude of the SRT (FTT) would be directly related to the magnitude of the displacement of the target due to kinematic response characteristics of the hand and arm.
To validate the study hypothesis, CL for both small and large displacements was calculated by subtracting the respective NL from the total SRT. Utilizing a repeated measures t-test analysis utilizing SPSS, a significant difference between CL (SD) and CL (LD) at p=0.696 was not observed.
Differentiation and quantification of individual components of CL and NL from a subject\'s total SRT was accomplished. An Apple iPad Pro was selected as a platform for our study because: (1) it is readily available, affordable, and programmable; (2) it meets a requirement for portability; and (3) it allows the modification/addition of test parameters to meet future needs. The ability to quantify the extent of cognitive and neuromuscular dysfunction in the TBI patient is an essential component of developing an effective treatment plan.

Motoneurons (MNs) control muscle activity by releasing the neurotransmitter acetylcholine (ACh) at the level of neuromuscular junctions. ACh is packaged into synaptic vesicles by the vesicular ACh transporter (VAChT), and disruptions in its release can impair muscle contraction, as seen in congenital myasthenic syndromes (CMS). Recently, VAChT gene mutations were identified in humans displaying varying degrees of myasthenia. Moreover, mice with a global deficiency in VAChT expression display several characteristics of CMS. Despite these findings, little is known about how a long-term decrease in VAChT expression in vivo affects MNs structure and function. Using Cre-loxP technology, we generated a mouse model where VAChT is deleted in select groups of MNs (mnVAChT-KD). Molecular analysis revealed that the VAChT deletion was specific to MNs and affected approximately 50% of its population in the brainstem and spinal cord, with alpha-MNs primarily targeted (70% in spinal cord). Within each animal, the cell body area of VAChT-deleted MNs was significantly smaller compared to MNs with VAChT preserved. Likewise, muscles innervated by VAChT-deleted MNs showed atrophy while muscles innervated by VAChT-containing neurons appeared normal. In addition, mnVAChT KD mice had decreased muscle strength, were hypoactive, leaner and exhibited kyphosis. This neuromuscular dysfunction was evident at 2 months of age and became progressively worse by 6 months. Treatment of mutants with a cholinesterase inhibitor was able to improve some of the motor deficits. As these observations mimic what is seen in CMS, this new line could be valuable for assessing the efficacy of potential CMS drugs.

Neuromuscular dysfunction is common in old age. Damaged cytoplasmic structures aggregate with aging, especially in post-mitotic cells like motor neurons. Autophagy is a ubiquitous cell process that aids in the clearance of damaged aggregates. Accordingly, we hypothesized that autophagy is impaired in old age, contributing to neuromuscular dysfunction via an effect in motor neurons. Autophagy flux may be impaired as a result of deficits in the initiation, elongation or degradation phases. Changes in the expression levels of core proteins necessary for each of the autophagy phases were evaluated by Western blotting in the cervical spinal cord (segments C2-C6 corresponding to the phrenic motor pool) of adult male and female mice at 6-, 18-, and 24-months of age (reflecting 100%, 90% and 75% survival, respectively). There was no evidence of an effect of age on the expression of the autophagy markers Beclin-1 (Becn-1; initiation), ATG7 and ATG5/12 complex (elongation) or LC3 (elongation/degradation). Reduced p62 expression (a marker of degradation) was evident in the cervical spinal cord of adult mice at 18-months compared to 24-months. Accordingly, expression of LC3 and p62 in motor neurons was analyzed using immunofluorescence and confocal microscopy in separate animals. LC3 and p62 immunoreactivity was evident in the gray matter with minimal expression in the white matter across all age groups. A mixed linear model with animal as a random effect was used to compare relative LC3 and p62 expression in motor neurons to gray matter across age groups. Expression of both LC3 and p62 was higher in choline acetyl transferase (ChAT)-positive motor neurons (~2-3 fold vs. gray matter). Across age groups, there were differences in the relative expression of LC3 (F2,12 = 7.59, p < 0.01) and p62 (F2,12 = 8.00, p < 0.01) in cervical motor neurons. LC3 expression in motor neurons increased ~20% by 24-months of age in both male and female mice. p62 expression in motor neurons increased ~70% by 18-months compared to 6-months with no further changes by 24-months of age in male mice. p62 expression did not change across age groups in female mice, and was ~20% higher than in males. Our findings highlight important changes in autophagy pathways that likely contribute to the development of aging-related neuromuscular dysfunction in mice. At 18-months of age, increased autophagosome clearance (reduced p62 expression) appears to be a global effect not restricted to motor neurons. By 24-months of age, increased expression of LC3 and p62 indicates impaired autophagy with autophagosome accumulation in cervical motor neurons.

Transcutaneous electrical muscle stimulation (TEMS) has been progressively used as add-on therapy to reduce muscle atrophy in adults unable to carry out active mobilization in the intensive care unit (ICU). There are no studies addressing TEMS in the pediatric ICU. Therefore, we decided to develop a scoping review, a type of knowledge synthesis, which unlike systematic review, identify gaps in the literature to aid the planning and commissioning of future research.
To provide current perspectives on the application of TEMS for combating pediatric intensive care unit acquired weakness (PICUAW).
Online databases were used to identify papers published 2006-2016, from which we selected those used musculoskeletal and cardiorespiratory performance as a primary or secondary outcome variable in participants under 18 years.
The publications reported six clinical trials from 218 outpatients with 9.5 ± 8 years old. There were differences in current modulation and duration of TEMS sessions, with a predominance of high intensity and short duration in which a muscle contraction is triggered. The main use of TEMS was in pediatric neurological disorders. TEMS was more effective when compared with SHAM on spasticity, bone mineral density, disability, and gait. One study regarding spine injury showed improvement in VO2 (P = 0.035) when combined cycling with TEMS.
TEMS was an effective and safe treatment for musculoskeletal impairments and cardiorespiratory performance in children with neurological disorders. Although the physiopathology is different in outpatients, an individualized protocol with TEMS might be promising for preventing PICUAW. Its effectiveness, however, deserves further investigation.

Abnormal expression and distribution of nicotinic acetylcholine receptors (nAChRs) in skeletal muscle caused by sepsis can lead to neuromuscular dysfunction. Here, we asked whether neural agrin regulates nAChRs to ameliorate muscle function, which could be associated with the agrin/muscle-specific kinase pathway.
Rats were subjected to cecal ligation and puncture (CLP) group, sham group, or control group to observe the alteration caused by sepsis. To verify the effect of improving function, rats were injected with agrin or normal saline intramuscularly after CLP. Electromyogram was used to measure neuromuscular function. Cytokines levels of serum and the expression of related proteins and mRNA were tested after treatment.
Compared with the rats in control or sham group, CLP-treated rats showed an acute inflammatory status and a reduction of neuromuscular dysfunction in tibialis anterior muscle, which was associated with abnormal expression in agrin/muscle-specific kinase pathway and increased expression of γ- and α7-nAChR. Exogenous agrin alleviated neuromuscular dysfunction and decreased the expression of γ- and α7-nAChR through agrin-related signaling pathway.
The decreased expression of agrin may lead to skeletal muscle dysfunction. Early enhancement of intramuscular agrin levels after sepsis may be a potential strategy for the treatment of sepsis-induced muscle dysfunction.

Intensive care unit-acquired weakness (ICU-AW) is associated with poorer outcome of critically ill patients. Microcirculatory changes and altered vascular permeability of skeletal muscles might contribute to the pathogenesis of ICU-AW. Muscular ultrasound (MUS) displays increased muscle echogenicity, although its pathogenesis is uncertain.
We investigated the combined measurement of serum and ultrasound markers to assess ICU-AW and clinical patient outcome.
Fifteen patients and five healthy controls were longitudinally assessed for signs of ICU-AW at study days 3 and 10 using a muscle strength sum score. The definition of ICU-AW was based on decreased muscle strength assessed by the muscular research council-sum score. Ultrasound echogenicity of extremity muscles was assessed using a standardized protocol. Serum markers of inflammation and endothelial damage were measured. The 3-month outcome was assessed on the modified Rankin scale.
ICU-AW was present in eight patients, and seven patients and the control subjects did not develop ICU-AW. The global muscle echogenicity score (GME) differed significantly between controls and patients (mean GME, 1.1 ± 0.06 vs. 2.3 ± 0.41; p = 0.001). Mean GME values significantly decreased in patients without ICU-AW from assessment 1 (2.30 ± 0.48) to assessment 2 (2.06 ± 0.45; p = 0.027), which was not observed in patients with ICU-AW. Serum levels of syndecan-1 at day 3 significantly correlated with higher GME values at day 10 (r = 0.63, p = 0.012). Furthermore, the patients\' GME significantly correlated with mRS at day 100 (r = 0.67, p = 0.013).
The combined use of muscular ultrasound and inflammatory biomarkers might be helpful to diagnose ICU-AW and to predict long-term outcome in critical illness.

Herpes simplex virus type 1 (HSV-1) is a widespread neurotropic pathogen responsible for a range of clinical manifestations. Inflammatory cell infiltrate is a common feature of HSV-1 infections and has been implicated in neurodegeneration. Therefore, viral recognition by innate immune receptors (i.e., TLR2) and the subsequent inflammatory response are now deemed key players in HSV-1 pathogenesis. In this study we infected with HSV-1 the enteric nervous system (ENS) of wild-type (WT) and TLR2 knock-out (TLR2ko) mice to investigate whether and how TLR2 participates in HSV-1 induced neuromuscular dysfunction. Our findings demonstrated viral specific transcripts suggestive of abortive replication in the ENS of both WT and TLR2ko mice. Moreover, HSV-1 triggered TLR2-MyD88 depend signaling in myenteric neurons and induced structural and functional alterations of the ENS. Gastrointestinal dysmotility was, however, less pronounced in TLR2ko as compared with WT mice. Interesting, HSV-1 caused up-regulation of monocyte chemoattractant protein-1 (CCL2) and recruitment of CD11b+ macrophages in the myenteric ganglia of WT but not TLR2ko mice. At the opposite, the myenteric plexuses of TLR2ko mice were surrounded by a dense infiltration of HSV-1 reactive CD3+CD8+INFγ+ lymphocytes. Indeed, depletion CD3+CD8+ cells by means of administration of anti-CD8 monoclonal antibody reduced neuromuscular dysfunction in TLR2ko mice infected with HSV-1. During HSV-1 infection, the engagement of TLR2 mediates production of CCL2 in infected neurons and coordinates macrophage recruitment. Bearing in mind these observations, blockage of TLR2 signaling could provide novel therapeutic strategies to support protective and specific T-cell responses and to improve neuromuscular dysfunction in pathogen-mediated alterations of the ENS.