In Situ Effects of Doxycycline on Neuromuscular Junction in Mice.

BACKGROUND: The antibacterial mechanism of doxycycline is known, but its effects on the nerve-muscle system are still not unclear. OBJECTIVE: The aim of the study was to combine molecular targets of the neuromuscular machinery using the in situ neuronal blocker effect of doxycycline, a semisynthetic second-generation tetracycline derivative, on mice neuromuscular preparations. METHODS: The effects of doxycycline were assessed on presynaptic, synaptic cleft, and postsynaptic neurotransmission, along with the muscle fiber, using the traditional myographic technique. Precisely, the effects of doxycycline were categorized into "all" or "nothing" effects depending on the concentration of doxycycline used; "all" was obtained with 4 µM doxycycline, and "nothing" was obtained with 1-3 µM doxycycline. The rationale of this study was to apply known pharmacological tools against the blocker effect of 4 µM doxycycline, such as F55-6 (Casearia sylvestris), CaCl2 (or Ca2+), atropine, neostigmine, polyethylene glycol (PEG 400), and d-Tubocurarine. The evaluation of cholinesterase enzyme activity and the diaphragm muscle histology were performed, and protocols on the neuromuscular preparation submitted to indirect or direct stimuli were complementary. RESULTS: Doxycycline does not affect cholinesterase activity nor causes damage to skeletal muscle diaphragm; it acts on ryanodine receptor, sarcolemmal membrane, and neuronal sodium channel with a postjunctional consequence due to the decreased availability of muscle nicotinic acetylcholine receptors. CONCLUSION: In conclusion, in addition to the neuronal blocker effect of doxycycline, we showed that doxycycline acts on multiple targets. It is antagonized by F55-6, a neuronal Na+-channel agonist, and Ca2+, but not by neostigmine.

The contemporary model of vertebral column joint dysfunction and impact of high-velocity, low-amplitude controlled vertebral thrusts on neuromuscular function.

PURPOSE: There is growing evidence that vertebral column function and dysfunction play a vital role in neuromuscular control. This invited review summarises the evidence about how vertebral column dysfunction, known as a central segmental motor control (CSMC) problem, alters neuromuscular function and how spinal adjustments (high-velocity, low-amplitude or HVLA thrusts directed at a CSMC problem) and spinal manipulation (HVLA thrusts directed at segments of the vertebral column that may not have clinical indicators of a CSMC problem) alters neuromuscular function. METHODS: The current review elucidates the peripheral mechanisms by which CSMC problems, the spinal adjustment or spinal manipulation alter the afferent input from the paravertebral tissues. It summarises the contemporary model that provides a biologically plausible explanation for CSMC problems, the manipulable spinal lesion. This review also summarises the contemporary, biologically plausible understanding about how spinal adjustments enable more efficient production of muscular force. The evidence showing how spinal dysfunction, spinal manipulation and spinal adjustments alter central multimodal integration and motor control centres will be covered in a second invited review. RESULTS: Many studies have shown spinal adjustments increase voluntary force and prevent fatigue, which mainly occurs due to altered supraspinal excitability and multimodal integration. The literature suggests physical injury, pain, inflammation, and acute or chronic physiological or psychological stress can alter the vertebral column's central neural motor control, leading to a CSMC problem. The many gaps in the literature have been identified, along with suggestions for future studies. CONCLUSION: Spinal adjustments of CSMC problems impact motor control in a variety of ways. These include increasing muscle force and preventing fatigue. These changes in neuromuscular function most likely occur due to changes in supraspinal excitability. The current contemporary model of the CSMC problem, and our understanding of the mechanisms of spinal adjustments, provide a biologically plausible explanation for how the vertebral column's central neural motor control can dysfunction, can lead to a self-perpetuating central segmental motor control problem, and how HVLA spinal adjustments can improve neuromuscular function.

Ischemic stroke-induced polyaxonal innervation at the neuromuscular junction is attenuated by robot-assisted mechanical therapy.

Ischemic stroke is a leading cause of disability world-wide. Mounting evidence supports neuromuscular pathology following stroke, yet mechanisms of dysfunction and therapeutic action remain undefined. The objectives of our study were to investigate neuromuscular pathophysiology following ischemic stroke and to evaluate the therapeutic effect of Robot-Assisted Mechanical massage Therapy (RAMT) on neuromuscular junction (NMJ) morphology. Using an ischemic stroke model in male rats, we demonstrated longitudinal losses of muscle contractility and electrophysiological estimates of motor unit number in paretic hindlimb muscles within 21 days of stroke. Histological characterization demonstrated striking pre- and postsynaptic alterations at the NMJ. Stroke prompted enlargement of motor axon terminals, acetylcholine receptor (AChR) area, and motor endplate size. Paretic muscle AChRs were also more homogenously distributed across motor endplates, exhibiting fewer clusters and less fragmentation. Most interestingly, NMJs in paretic muscle exhibited increased frequency of polyaxonal innervation. This finding of increased polyaxonal innervation in stroke-affected skeletal muscle suggests that reduction of motor unit number following stroke may be a spurious artifact due to overlapping of motor units rather than losses. Furthermore, we tested the effects of RAMT - which we recently showed to improve motor function and protect against subacute myokine disturbance - and found significant attenuation of stroke-induced NMJ alterations. RAMT not only normalized the post-stroke presentation of polyaxonal innervation but also mitigated postsynaptic expansion. These findings confirm complex neuromuscular pathophysiology after stroke, provide mechanistic direction for ongoing research, and inform development of future therapeutic strategies. SIGNIFICANCE: Ischemic stroke is a leading contributor to chronic disability, and there is growing evidence that neuromuscular pathology may contribute to the impact of stroke on physical function. Following ischemic stroke in a rat model, there are progressive declines of motor unit number estimates and muscle contractility. These changes are paralleled by striking pre- and postsynaptic maladaptive changes at the neuromuscular junction, including polyaxonal innervation. When administered to paretic hindlimb muscle, Robot-Assisted Mechanical massage Therapy - previously shown to improve motor function and protect against subacute myokine disturbance - prevents stroke-induced neuromuscular junction alterations. These novel observations provide insight into the neuromuscular response to cerebral ischemia, identify peripheral mechanisms of functional disability, and present a therapeutic rehabilitation strategy with clinical relevance.

Functional, physiological and subjective responses to concurrent neuromuscular electrical stimulation (NMES) exercise in adult cancer survivors: a controlled prospective study.

The primary aim of this study was to investigate the functional, physiological and subjective responses to NMES exercise in cancer patients. Participants with a cancer diagnosis, currently undergoing treatment, and an had an Eastern Cooperative Oncology Group (ECOG) performance status (ECOG) of 1 and 2 were recommended to participate by their oncologist. Following a 2-week, no-NMES control period, each participant was asked to undertake a concurrent NMES exercise intervention over a 4-week period. Functional muscle strength [30 s sit-to-stand (30STS)], mobility [timed up and go (TUG)], exercise capacity [6-min walk test (6MWT)] and health related quality of life (HR-QoL) were assessed at baseline 1 (BL1), 2-week post control (BL2) and post 4-week NMES exercise intervention (POST). Physiological and subjective responses to LF-NMES were assessed during a 10-stage incremental session, recorded at BL2 and POST. Fourteen participants [mean age: 62 years (10)] completed the intervention. No adverse events were reported. 30STS (+ 2.4 reps, p = .007), and 6MWT (+ 44.3 m, p = .028) significantly improved after the intervention. No changes in TUG or HR-QoL were observed at POST. Concurrent NMES exercise may be an effective exercise intervention for augmenting physical function in participants with cancer and moderate and poor functional status. Implications for cancer survivors: By allowing participants to achieve therapeutic levels of exercise, concurrent NMES may be an effective supportive intervention in cancer rehabilitation.

Improved Human Muscle Biopsy Method To Study Neuromuscular Junction Structure and Functions with Aging.

Reduced mobility and physical independence of elders has emerged as a major clinical and public health priority with extended life expectancy. The impact of the neuromuscular function on muscle activity and properties has emerged as a critical factor influencing the progress and outcome of muscle changes with aging. However, very little is known about the neuromuscular junctions (NMJs) in humans, in part due to technical constraints limiting the access to healthy, fresh neuromuscular tissue. Here, we describe a method, called Biopsy using Electrostimulation for Enhanced NMJ Sampling (BeeNMJs) that improves the outcome of muscle biopsies. We used local cutaneous stimulation to identify the area enriched with NMJs for each participant at the right Vastus lateralis (VL). The needle biopsy was then performed in proximity of that point. The BeeNMJs procedure was safe for the participants. We observed NMJs in 53.3% of biopsies in comparison with only 16.7% using the traditional method. Furthermore, we observed an average of 30.13 NMJs per sample compared to only 2.33 for the traditional method. Importantly, high-quality neuromuscular material was obtained whereby pre-, postsynaptic, and glial elements were routinely labeled, simultaneously with myosin heavy chain type I. The BeeNMJs approach will facilitate studies of NMJs, particularly in human disease or aging process.

Creatine supplementation improves performance above critical power but does not influence the magnitude of neuromuscular fatigue at task failure.

NEW FINDINGS: What is the central question of this study? Does the magnitude of neuromuscular fatigue depend on the amount of work done (W') at task failure when cycling above critical power (CP)? What is the main finding and its importance? Creatine supplementation increases W' and enhances supra-CP performance, but induces similar magnitudes of neuromuscular fatigue at task failure compared to placebo. Increased W' does not lead to higher levels of neuromuscular fatigue. This supports the notion of a critical level of neuromuscular fatigue at task failure and challenges a direct causative link between W' depletion and neuromuscular fatigue. ABSTRACT: The present study examined the effect of creatine supplementation on neuromuscular fatigue and exercise tolerance when cycling above critical power (CP). Eleven males performed an incremental cycling test with four to five constant-load trials to task failure (TTF) to obtain asymptote (CP) and curvature constant (W') of the power-duration relationship, followed by three constant-load supra-CP trials: (1) one TTF following placebo supplementation (PLA); (2) one TTF following creatine supplementation (CRE); and (3) one trial of equal duration to PLA following creatine supplementation (ISO). Neuromuscular assessment of the right knee extensors was performed pre- and post-exercise to measure maximal voluntary contraction (MVC), twitch forces evoked by single (Qpot ) and paired high- (PS100) and low- (PS10) frequency stimulations and voluntary activation. Creatine supplementation increased TTF in CRE vs. PLA by ∼11% (P = 0.017) and work done above CP by ∼10% (P = 0.015), with no difference (P > 0.05) in reductions in MVC (-24 ± 8% vs. -20 ± 9%), Qpot (-39 ± 13% vs. -32 ± 14%), PS10 (-42 ± 14% vs. -36 ± 13%), PS100 (-25 ± 10% vs. -18 ± 12%) and voluntary activation (-7 ± 8% vs. -5 ± 7%). No significant difference was found between ISO and either PLA or CRE (P > 0.05). These findings suggest similar levels of neuromuscular fatigue can be found following supra-CP cycling despite increases in performance time and amount of work done above CP, supporting the notion of a critical level of neuromuscular fatigue and challenging a direct causative link between W' depletion and neuromuscular fatigue.

The Effects of Tai Chi Chuan Versus Core Stability Training on Lower-Limb Neuromuscular Function in Aging Individuals with Non-Specific Chronic Lower Back Pain.

Objectives: For this paper, we aimed to investigate the effects of Tai Chi Chuan (TCC) versus the Core Stability Training (CST) program on neuromuscular function (NF) in the lower extremities among aging individuals who suffered from non-specific chronic lower back pain (NLBP). Regarding the design, during a 12-week intervention, a single-blinded randomized controlled trial was used to compare two intervention groups with a control group on the parameters of NF. Methods: Forty-three Chinese community-dwellers were randomly assigned into two intervention groups (three sessions per week, with each session lasting 60 min in TCC and CST) and a control group. The patient-based Visual Analogue Scale (VAS) was used to measure the level of perceived pain, while parameters of NF as primary outcomes were measured by the Biodex System 3 Isokinetic Dynamometer. Results: For the knee joint, we observed significant differences in the endurance of left extension at a speed of 60°/s: (1) between TCC and control groups (p < 0.01); (2) between CST and control groups (p < 0.01). For the ankle joint, significant differences between CST and control groups were observed on the peak torque of left dorsiflexion (p < 0.05) and the endurance of the left plantar flexion at a speed of 60°/s (p < 0.05). In addition, we observed a significant difference between TCC and control groups in the endurance of the right plantar flexion (p < 0.05). Conclusions: Chen-style TCC and CST were found to have protective effects on NF in aging individuals with NLBP, while alleviating non-specific chronic pain.

Neuromuscular efficiency in fibromyalgia is improved by hyperbaric oxygen therapy: looking inside muscles by means of surface electromyography.

OBJECTIVES: Neuromuscular efficiency (NME) is impaired in fibromyalgia (FM). Hyperbaric oxygen therapy (HBOT) is a medical treatment using 100% of oxygen through an oxygen mask. HBOT in FM induces changes in cortical excitability and a secondary reduction in pain and muscle fatigue. However, there are still no direct data indicating changes in muscle fatigue. The aim of this study was to assess whether the reduction in muscle fatigue so far attributed to a central effect of HBOT can be directly detected by means of non-invasive sEMG as a change in NME. METHODS: The study was an observational longitudinal study on changes in NME induced by 20 sessions of HBOT at 2.4 atmosphere, in 22 patients with FM (3M; 19F) (age 49.8±9.5; height 164.7±7.5; weight 63.8±12.7). sEMG was recorded in single differential configuration from the biceps brachii muscle during the 30-second fatiguing contractions using linear arrays of eight adhesive electrodes. RESULTS: Evaluations made before and immediately after the first session showed that maximal strength did not change (T0 49±20 N, T1 49±19 N, p=0.792), thus suggesting that HBOT did not induce muscle fatigue or potentiation. After 20 sessions of HBOT, NME increased from 1.6±1.1 to 2.1±0.8 (p=0.050), whereas maximal strength, EMG amplitude and muscle fibre CV did not change. CONCLUSIONS: HBOT did not improve muscle strength or change muscle fibre content, but improved the ability of the central motor command to generate the same effort (MVC) with fewer recruited fibres. Our sEMG findings underlined a modified central mechanism related to fibre type recruitment order, thus suggesting that muscle fatigue is not primarily a muscular problem, as also demonstrated by other authors with different methods.

Effects of the hybrid of neuromuscular electrical stimulation and noxious thermal stimulation on upper extremity motor recovery in patients with stroke: a randomized controlled trial.

BACKGROUND: Neuromuscular electrical stimulation (NMES) and noxious thermal stimulation (NTS) have been developed and incorporated in stroke rehabilitation. OBJECTIVE: This study aimed to compare the effects of NMES, NTS, and the hybrid of NMES and NTS ("Hybrid") on motor recovery of upper extremity (UE) for patients with stroke. METHODS: We conducted a prospective, single-blind randomized controlled trial with concealed allocation. Forty-three patients with chronic stroke (onset >6 months) were randomly assigned to three groups (NMES, NTS, and "Hybrid"). In addition to conventional rehabilitation, participants received 30 min of NMES or 30 min of NTS or 15 min of NTS followed by 15 min of NMES. The treatment period was 8 weeks, 3 days/week, 30 min/time. The UE subscale of Fugl-Meyer assessment (UE-FMA, the primary outcome), Motricity index, modified Ashworth scale, and Barthel index were administered by a blinded assessor at baseline, posttreatment, and one-month follow-up. RESULTS: Most of the participants had mild-to-moderate disability in activity of daily living. No significant differences in the outcome measures at posttreatment and one-month follow-up were found among the NMES group (n = 13), NTS group (n = 13), and the hybrid of NMES and NTS group (n = 17). However, significant score changes in UE-FMA (p < 0.025) from baseline to posttreatment and one-month follow-up were found for the "Hybrid" group. CONCLUSIONS: This study reveals that the hybrid of NMES and NTS therapy appears to be beneficial to UE recovery after stroke but is not superior to NMES or NTS alone.

Train-of-four guard-controlled sugammadex reversal in patients with multiple sclerosis.

OBJECTIVE: Multiple sclerosis (MS) is a chronic inflammatory and demyelinating autoimmune disease of the central nervous system. It is known that the disease, which is manifested by a wide variety of symptoms, may exacerbate after anesthesia and show different responses to muscle relaxants in the normal population. It is planned to measure train-of-four (TOF) values of MS patients to be operated under general anesthesia before sugammadex application. MATERIALS AND METHODS: With the approval of the local ethics committee of the University of Health Sciences Bagcilar Training and Research Hospital and with written consents of participants, we anesthetized ten patients (from April 2014 to March 2017) with MS and ten American Society of Anesthesiologists I-III patients without MS. Neuromuscular conduction was assessed by the acceleromyometric method using a TOF-Guard apparatus (Organon, Holland). The patients were extubated after recovery of TOF higher than 0.9. The primary efficacy variable was the time from the start of administration of sugammadex to recovery of the TOF ratio to 0.9. RESULTS: The demographic characteristics of both groups, the type and duration of surgery and anesthesia applied, and the temperature of the operation room were similar. Similar characteristics of both groups were of concern for postoperative residual paralysis, and therefore we did not notice any difference between time to TOF> 90/s for both groups. CONCLUSION: Sugammadex and TOF patients will increase patient safety in general anesthesia practice.

Wide-pulse, high-frequency, low-intensity neuromuscular electrical stimulation has potential for targeted strengthening of an intrinsic foot muscle: a feasibility study.

BACKGROUND: Strengthening the intrinsic foot muscles is a poorly understood and largely overlooked area. In this study, we explore the feasibility of strengthening m. abductor hallucis (AH) with a specific paradigm of neuromuscular electrical stimulation; one which is low-intensity in nature and designed to interleave physiologically-relevant low frequency stimulation with high-frequencies to enhance effective current delivery to spinal motoneurones, and enable a proportion of force produced by the target muscle to be generated from a central origin. We use standard neurophysiological measurements to evaluate the acute (~ 30 min) peripheral and central adaptations in healthy individuals. METHODS: The AH in the dominant foot of nine healthy participants was stimulated with 24 × 15 s trains of square wave (1 ms), constant current (150% of motor threshold), alternating (20 Hz-100 Hz) neuromuscular electrical stimulation interspersed with 45 s rest. Prior to the intervention, peripheral variables were evoked from the AH compound muscle action potential (Mwave) and corresponding twitch force in response to supramaximal (130%) medial plantar nerve stimulation. Central variables were evoked from the motor evoked potential (MEP) in response to suprathreshold (150%) transcranial magnetic stimulation of the motor cortex corresponding to the AH pathway. Follow-up testing occurred immediately, and 30 min after the intervention. In addition, the force-time-integrals (FTI) from the 1st and 24th WPHF trains were analysed as an index of muscle fatigue. All variables except FTI (T-test) were entered for statistical analysis using a single factor repeated measures ANOVA with alpha set at 0.05. RESULTS: FTI was significantly lower at the end of the electrical intervention compared to that evoked by the first train (p < 0.01). Only significant peripheral nervous system adaptations were observed, consistent with the onset of low-frequency fatigue in the muscle. In most of these variables, the effects persisted for 30 min after the intervention. CONCLUSIONS: An acute session of wide-pulse, high-frequency, low-intensity electrical stimulation delivered directly to abductor hallucis in healthy feet induces muscle fatigue via adaptations at the peripheral level of the neuromuscular system. Our findings would appear to represent the first step in muscle adaptation to training; therefore, there is potential for using WPHF for intrinsic foot muscle strengthening.

Efficiency of neuromuscular electrical stimulation: A comparison of elicited force and subject tolerance using three electrical waveforms.

OBJECTIVE: Efficacy of neuromuscular electrical stimulation (NMES) is limited by the discomfort of electrically elicited contractions. Most studies of tolerance to NMES have examined stimulation to maximal tolerance. NMES efficiency is the amount of elicited force at a specific level of tolerance. This study is the first to describe and examine such. DESIGN: A repeated measures design was used. Electrically elicited force (EEF) was measured using three waveforms: burst-modulated alternating current (BMAC), pulsed current (PC), and burst-modulated pulsed current (BMPC). EEF at a tolerance rating of 5/10 on a visual analog scale (VAS) was recorded. The dependent variables were EEF up to 5/10 VAS, current amplitude at 5/10, and percent maximal isometric force at 5/10. RESULTS: EEF and percent maximal voluntary isometric force were significantly greater with BMPC versus BMAC (p = 0.001 and 0.004). No differences were noted between PC and BMAC or BMPC and PC. Amplitude was significantly greater with BMAC compared to BMPC and PC (p = 0.003 and 0.015). No difference in amplitude was noted between PC and BMPC. CONCLUSION: For the same level of discomfort, BMPC yielded one-third greater muscle force than BMAC and at a lesser current amplitude. These data evidence a greater efficiency for BMPC than BMAC.

Neuromuscular electrical stimulation reduces sludge in the popliteal vein.

BACKGROUND: The common peroneal nerve stimulator (CPNS) is a UK-approved device for reducing venous thromboembolism (VTE) risk. It resembles a wrist watch and is placed over the common peroneal nerve, discharging electricity at a rate of 1 impulse/s. It has been presumed that as blood flow slows, erythrocytes aggregate into ultrasound-detectable echogenic particles, described as venous sludge. The aim of the study was to determine whether the CPNS reduces venous sludge by using an ultrasound-derived gray-scale (0-255) venous sludge index (VSI). METHODS: Twenty-five healthy volunteers had their right popliteal vein video recorded using B-mode ultrasound at 22 frames/s in longitudinal and transverse views, standing and lying. This was performed first with the CPNS off and then with the CPNS on. The CPNS impulse intensity used was set from 1 to 7 for each individual, and the level was sufficient to cause an outward jerking movement of the foot. A single frame of the possible 154 frames, lasting 7 seconds, was selected using a random number generator for the image analysis. The "brightness" of the erythrocyte aggregates (pixels) within a circular sampling area was quantified using the VSI. The brighter the sample, the greater the sludge. RESULTS: Values are expressed as median (interquartile range). On standing with the device off, there was a significantly higher VSI (P < .0005) compared with lying (longitudinal view, 27.7 [18.8-41.4] vs 11.7 [5.5-17.5]; transverse view, 20.7 [13.6-32.2] vs 11.4 [6.3-15.9]). Activation of the CPNS significantly reduced all the VSI values (P < .0005) shown (longitudinal view, 2 [1.1-3.2] and 1.5 [0.5-3.1]; transverse view, 1.1 [0.6-2.7] and 0.8 [0.5-2.1]). CONCLUSIONS: The CPNS device significantly reduces venous sludge within the popliteal vein irrespective of whether the subject is standing or lying down or of the longitudinal or transverse position of the ultrasound transducer. The principal mode of action of the device in the claim that it may reduce venous thromboembolism risk may be through a reduction of venous sludge. However, the relationship between erythrocyte aggregation, venous stasis, and venous thromboembolism risk requires more investigation.

[Claude Bernard and nicotinic receptors: from the neuromuscular junction to tobacco weaning]. / Claude Bernard et les récepteurs nicotiniques : de la jonction neuromusculaire au sevrage tabagique.

Claude Bernard (1813-1878) was fascinated by the pharmacological mechanisms of poisons. In particular, using a huge amount of ingenious and robust experiments, he demonstrated the peripheral toxic action of the natural compound curare. His work generated controversies in a period where scientific methodology and technical development followed the progression of concepts and ideas. From his intense debates with Albert Vulpian emerged the location of curare's toxicity at the neuromuscular junction. These two fascinating scientists could not imagine how important were these discoveries which allowed John Langley to propose the concept of receptor early in the 20th century. At the same time, the German immunologist Paul Ehrlich suggested that these receptors could be targeted by so-called "magic bullets", i.e., drugs that act on receptors, in order to treat patients. The molecular substrate of curare's activity was identified many years later as the nicotinic receptor of the motor end-plate. We now have curare molecules belonging to various chemical families that block receptors during anaesthesia. Suggamadex is the antidote for two of them, a drug that Claude Bernard perhaps dreamt of. We also have the recently marketed varenicline that acts as a partial agonist of nicotinic receptors in the central nervous system to treat patients from tobacco addiction. This rich story shows that biomedical research needs collaborations, imagination, perspicacity but also all results that it can have many years later, therefore challenging researchers about consequences of their discoveries.

Bioreactor model of neuromuscular junction with electrical stimulation for pharmacological potency testing.

In vitro models of the neuromuscular junction (NMJ) are emerging as a valuable tool to study synaptogenesis, synaptic maintenance, and pathogenesis of neurodegenerative diseases. Many models have previously been developed using a variety of cell sources for skeletal muscle and motoneurons. These models can advanced by integrating beneficial features of the native developmental milieu of the NMJ. We created a functional in vitro model of NMJ by bioreactor cultivation of transdifferentiated myocytes and stem cell-derived motoneurons, in the presence of electrical stimulation. In conjunction with a coculture medium, electrical stimulation resulted in improved maturation and function of motoneurons and myocytes, as evidenced by mature cellular structures, increased expression of neuronal and muscular genes, clusterization of acetylcholine receptors (AChRs) in the vicinity of motoneurons, and the response to glutamate stimulation. To validate the model and demonstrate its utility for pharmacological testing, we documented the potency of drugs that affect key pathways during NMJ signal transduction: (i) acetylcholine (ACh) synthesis, (ii) ACh vesicular storage, (iii) ACh synaptic release, (iv) AChR activation, and (v) ACh inactivation in the synaptic cleft. The model properly responded to the drugs in a concentration-dependent manner. We thus propose that this in vitro model of NMJ could be used as a platform in pharmacological screening and controlled studies of neuromuscular diseases.

Effects of neuromuscular electrical stimulation on cytokines in peripheral blood for healthy participants: a prospective, single-blinded Study.

INTRODUCTION: The effect of exercise on cytokines may improve muscle strength. Neuromuscular electrical stimulation (NMES) is a muscle-preserving therapy that benefits patients unable to participate in active exercise. How NMES alters cytokines is unclear. The aim of this study was to study the effects of 1 NMES session on cytokines associated with protein metabolism during exercise. METHODS: We evaluated the effects of NMES on IL-1, IL-6, IL-10 and TNF-α levels in peripheral blood. Participants received NMES to bilateral lower extremity muscles (quadriceps, tibialis anterior, gastrocnemius) for 30 min. Blood samples immediately pre- and post-NMES were drawn at 15-min intervals to 2-h follow-up, and the mean values of pre-NMES levels were compared to peak and trough post-NMES levels. For cytokines with significant changes, we conducted a repeated-measures linear regression analysis. We also measured post-NMES lactate and creatine kinase levels. RESULTS: We enrolled nine eligible participants. There was a significant increase in peak IL-6 from the mean pre-NMES value [0·65 (0·89) to 1·04 (0·89) pg ml-1 , P = 0·001] and a significant decrease in trough IL-1 [0·08 (0·07) to 0·02 (0·02) pg ml-1 , P = 0·041] and TNF-α [2·42 (0·54) to 2·16 (0·59) pg ml-1 , P = 0·021]. In repeated-measures regression analysis, we identified significantly higher mean IL-6 values throughout the full 120 min post-NMES period, and a significantly higher mean IL-1 value at 30 min post-NMES. There were no significant differences in peak IL-10, trough IL-6, lactate, or creatine kinase values. CONCLUSIONS: In nine healthy humans, 30 min of NMES was temporally associated with changes in cytokines similar to the effects of active exercise and may mediate NMES' observed effects on reducing muscle weakness.

Optimal duration of ultra low frequency-transcutaneous electrical nerve stimulation (ULF-TENS) therapy for muscular relaxation in neuromuscular occlusion: A preliminary clinical study.

OBJECTIVES: The primary aim of this work was to determine the duration of ultra-low-frequency transcutaneous electrical nerve stimulation (ULF-TENS) application necessary to achieve sufficient relaxation of the masticatory muscles. A secondary aim was to analyze the influence of stimulation on muscle relaxation in pathological subjects and determine whether ULF-TENS has a noteworthy impact on muscle relaxation. METHODS: Sixteen adult subjects with temporomandibular disorders (TMD) and muscle pain and a group of four control subjects were included in this study. ULF-TENS was applied, and muscular activities of the masseter, temporal, and sternocleidomastoid muscles (SCM) were recorded for 60 min. RESULTS: Significant relaxation was achieved in the TMD group from 20, 40, and 60 min for the temporal, masseter, and SCM muscles (p < 0.05), respectively. Maximum relaxation was achieved in 12.5% of the subjects after 20 min, in a further 12.5% after 40 min, and in the remaining 75% after 60 min. Significant relaxation was achieved in the control group from 20 to 40 min for the masseter and temporal muscles, respectively (p < 0.05). DISCUSSION: Taken together, the results suggest that an ideal ULF-TENS application would last 40 min to obtain sufficient muscle relaxation both in patients with masticatory system disorders and healthy subjects, a time constraint that is consistent with everyday clinical practice.

A new therapeutic application of brain-machine interface (BMI) training followed by hybrid assistive neuromuscular dynamic stimulation (HANDS) therapy for patients with severe hemiparetic stroke: A proof of concept study.

BACKGROUND: Hybrid assistive neuromuscular dynamic stimulation (HANDS) therapy improved paretic upper extremity motor function in patients with severe to moderate hemiparesis. We hypothesized that brain machine interface (BMI) training would be able to increase paretic finger muscle activity enough to apply HANDS therapy in patients with severe hemiparesis, whose finger extensor was absent. OBJECTIVE: The aim of this study was to assess the efficacy of BMI training followed by HANDS therapy in patients with severe hemiparesis. METHODS: Twenty-nine patients with chronic stroke who could not extend their paretic fingers were participated this study. We applied BMI training for 10 days at 40 min per day. The BMI detected the patients' motor imagery of paretic finger extension with event-related desynchronization (ERD) over the affected primary sensorimotor cortex, recorded with electroencephalography. Patients wore a motor-driven orthosis, which extended their paretic fingers and was triggered with ERD. When muscle activity in their paretic fingers was detected with surface electrodes after 10 days of BMI training, we applied HANDS therapy for the following 3 weeks. In HANDS therapy, participants received closed-loop, electromyogram-controlled, neuromuscular electrical stimulation (NMES) combined with a wrist-hand splint for 3 weeks at 8 hours a day. Before BMI training, after BMI training, after HANDS therapy and 3month after HANDS therapy, we assessed Fugl-Meyer Assessment upper extremity motor score (FMA) and the Motor Activity Log14-Amount of Use (MAL-AOU) score. RESULTS: After 10 days of BMI training, finger extensor activity had appeared in 21 patients. Eighteen of 21 patients then participated in 3 weeks of HANDS therapy. We found a statistically significant improvement in the FMA and the MAL-AOU scores after the BMI training, and further improvement was seen after the HANDS therapy. CONCLUSION: Combining BMI training with HANDS therapy could be an effective therapeutic strategy for severe UE paralysis after stroke.

Effect of neuromuscular electrical stimulation on motor cortex excitability upon release of tonic muscle contraction.

The aim of the present study was to investigate the neurophysiological triggers underlying muscle relaxation from the contracted state, and to examine the mechanisms involved in this process and their subsequent modification by neuromuscular electrical stimulation (NMES). Single-pulse transcranial magnetic stimulation (TMS) was used to produce motor-evoked potentials (MEPs) and short-interval intracortical inhibition (SICI) in 23 healthy participants, wherein motor cortex excitability was examined at the onset of voluntary muscle relaxation following a period of voluntary tonic muscle contraction. In addition, the effects of afferent input on motor cortex excitability, as produced by NMES during muscle contraction, were examined. In particular, two NMES intensities were used for analysis: 1.2 times the sensory threshold and 1.2 times the motor threshold (MT). Participants were directed to execute constant wrist extensions and to release muscle contraction in response to an auditory "GO" signal. MEPs were recorded from the flexor carpi radialis (FCR) and extensor carpi radialis (ECR) muscles, and TMS was applied at three different time intervals (30, 60, and 90 ms) after the "GO" signal. Motor cortex excitability was greater during voluntary ECR and FCR relaxation using high-intensity NMES, and relaxation time was decreased. Each parameter differed significantly between 30 and 60 ms. Moreover, in both muscles, SICI was larger in the presence than in the absence of NMES. Therefore, the present findings suggest that terminating a muscle contraction triggers transient neurophysiological mechanisms that facilitate the NMES-induced modulation of cortical motor excitability in the period prior to muscle relaxation. High-intensity NMES might facilitate motor cortical excitability as a function of increased inhibitory intracortical activity, and therefore serve as a transient trigger for the relaxation of prime mover muscles in a therapeutic context.