Passive Lengthening Research Articles

Myosin in autoinhibited off state(s), stabilized by mavacamten, can be recruited in response to inotropic interventions

Mavacamten is a FDA-approved small-molecule therapeutic designed to regulate cardiac function at the sarcomere level by selectively but reversibly inhibiting the enzymatic activity of myosin. It shifts myosin toward ordered off states close to the thick filament backbone. It remains elusive whether these myosin heads in the off state(s) can be recruited in response to physiological stimuli when required to boost cardiac output. We show that cardiac myosins stabilized in these off state(s) by mavacamten are recruitable by 1) Ca2+, 2) increased chronotropy [heart rate (HR)], 3) stretch, and 4) β-adrenergic (β-AR) stimulation, all known physiological inotropic interventions. At the molecular level, we show that Ca2+ increases myosin ATPase activity by shifting mavacamten-stabilized myosin heads from the inactive super-relaxed state to the active disordered relaxed state. At the myofilament level, both Ca2+ and passive lengthening can shift mavacamten-ordered off myosin heads from positions close to the thick filament backbone to disordered on states closer to the thin filaments. In isolated rat cardiomyocytes, increased stimulation rates enhanced shortening fraction in mavacamten-treated cells. This observation was confirmed in vivo in telemetered rats, where left-ventricular dP/dtmax, an index of inotropy, increased with HR in mavacamten-treated animals. Finally, we show that β-AR stimulation in vivo increases left-ventricular function and stroke volume in the setting of mavacamten. Our data demonstrate that the mavacamten-promoted off states of myosin in the thick filament are at least partially activable, thus preserving cardiac reserve mechanisms.

Coronal As Well As Sagittal Fascicle Dynamics Can Bring About a Gearing Effect in Muscle Elongation by Passive Lengthening.

The amount of muscle belly elongation induced by passive lengthening is often assumed to be equal to that of fascicles. But these are different if fascicles shorter than the muscle belly rotate around their attachment sites. Such discrepancy between fascicles and muscle belly length changes can be considered as gearing. As the muscle fascicle arrangement is 3D, the fascicle rotation by passive lengthening may occur in the coronal as well as the sagittal planes. Here we examined the fascicle 3D dynamics and resultant gearing during passive elongation of human medial gastrocnemius in vivo . For 16 healthy adults, we reconstructed fascicles three-dimensionally using diffusion tensor imaging and evaluated the change in fascicle length and angles in the sagittal and coronal planes during passive ankle dorsiflexion (from 20° plantar flexion to 20° dorsiflexion). Whole muscle belly elongation during passive ankle dorsiflexion was 38% greater than the fascicle elongation. Upon passive lengthening, the fascicle angle in the sagittal plane in all regions (-5.9°) and that in the coronal plane in the middle-medial (-2.7°) and distal-medial (-4.3°) regions decreased significantly. Combining the fascicle coronal and sagittal rotation significantly increased the gearing effects in the middle-medial (+10%) and distal-medial (+23%) regions. The gearing effect by fascicle sagittal and coronal rotations corresponded to 26% of fascicle elongation, accounting for 19% of whole muscle belly elongation. Fascicle rotation in the coronal and sagittal planes is responsible for passive gearing, contributing to the whole muscle belly elongation. Passive gearing can be favorable for reducing fascicle elongation for a given muscle belly elongation.

Regulation of primary afferent depolarization and homosynaptic post-activation depression during passive and active lengthening, shortening and isometric conditions.

This study aimed to determine whether the modulation of primary afferent depolarization (PAD) and homosynaptic post-activation depression (HPAD) are involved in the lower efficacy of Ia-afferent-α-motoneuron transmission commonly observed during lengthening compared to isometric and shortening conditions. 15 healthy young individuals participated in two experimental sessions dedicated to measurement in passive and active muscle states, respectively. In each session, PAD, HPAD and the efficacy of Ia-afferent-α-motoneuron transmission were evaluated during lengthening, shortening and isometric conditions. PAD was evaluated with D1 inhibition technique. Posterior tibial nerve stimulation was used to study HPAD and the efficacy of the Ia-afferent-α-motoneuron transmission through the recording of the soleus Hoffmann reflex (H reflex). PAD was increased in lengthening than shortening (11.2%) and isometric (12.3%) conditions regardless of muscle state (P < 0.001). HPAD was increased in lengthening than shortening (5.1%) and isometric (4.2%) conditions in the passive muscle state (P < 0.05), while no difference was observed in the active muscle state. H reflex was lower in lengthening than shortening (-13.2%) and isometric (-9.4%) conditions in both muscle states (P < 0.001). These results highlight the specific regulation of PAD and HPAD during lengthening conditions. However, the differences observed during passive lengthening compared to shortening and isometric conditions seem to result from an increase in Ia-afferent discharge, while the variations highlighted during active lengthening might come from polysynaptic descending pathways involving supraspinal centres that could regulate PAD mechanism.

Myosin in autoinhibited off state(s), stabilized by mavacamten, can be recruited via inotropic effectors.

Mavacamten is a novel investigational small molecule designed to regulate cardiac function at the sarcomere level by selectively but reversibly inhibiting the enzymatic activity of myosin. It shifts myosin towards ordered off states close to the thick filament backbone. However, the likelihood of recruiting these mavacamten-induced, off-state myosin heads upon increasing physiological demands of the heart has not been systematically explored. Here, we show that cardiac myosins stabilized in these off state(s) by mavacamten are recruitable by 1) Ca2+, 2) stretch and 3) β-adrenergic receptor (β-AR) stimulation, three inotropic effectors. At the molecular level, we find that Ca2+ increases myosin ATPase activity by shifting mavacamten-stabilized myosin heads from the inactive super-relaxed (SRX) state to the active disordered relaxed (DRX) state. At the sarcomere level, both Ca2+ and passive lengthening can shift mavacamten-ordered off myosin heads from positions close to the thick filament backbone to disordered on states closer to the thin filaments. And finally, in vivo, we show that β-AR stimulation can increase left-ventricular function and stroke volume in the setting of mavacamten. In conclusion, these data demonstrate that the mavacamten-promoted off states of myosin in the thick filament are activable, thus leading to the preservation of the cardiac reserve. These results not only provide a potential mechanistic explanation for the clinical observation of increased peak oxygen uptake (pVO2) with exercise in HCM patients receiving mavacamten but also lay out a fresh outlook on the safety and reversibility of this drug. This project is supported by grant P30 GM138395 from the NIGMS of the NIH; the Ministry of Education, Science and Technological Development of the Republic of Serbia through Contracts No. 451-03-68/2022-14/200378; MyoKardia Inc., a wholly owned subsidiary of Bristol Myers Squibb.

Reliability of shear wave elastography for the assessment of gastrocnemius fascia elasticity in healthy individual

The mechanical properties of the deep fascia, particularly their stiffness, strongly affect the development of muscle pathologies (such as compartment syndrome) and the action of the muscles. However, the mechanical characteristics of the deep muscular fascia are still not clearly understood. The present study focuses on examining the reliability of ultrasonic shear wave elastography (USWE) devices in quantifying the shear modulus of the gastrocnemius fascia in healthy individuals, particularly their ability to measure the shear modulus of the deep fascia of the gastrocnemius during ankle dorsiflexion. Twenty-one healthy males (age: 21.48 ± 1.17 years) participated in the study. Using USWE, the shear moduli of the medial gastrocnemius fascia (MGF) and lateral gastrocnemius fascia (LGF) were quantified at different angles during passive lengthening. The two operators took turns measuring each subject’s MGF and LGF over a 1-h period, and operator B took an additional measurement 2 h later. For the intra-operator test, the same subjects were measured again at the same time of day 5 days later. Both the intrarater [intraclass correlation coefficient (ICC) = 0.846–0.965)] and interrater (ICC = 0.877–0.961) reliability values for measuring the shear moduli of the MGF and LGF were rated as excellent; the standard error of the mean (SEM) was 3.49 kPa, and the minimal detectable change (MDC) was 9.68 kPa. Regardless of the ankle angle, the shear moduli of the LGF were significantly greater than that of the MGF (p < 0.001). Significant increases in the shear moduli of both the MGF and the LGF were observed in the neutral position compared to the relaxed position. These results indicate that USWE is a reliable technique to assess the shear modulus of the gastrocnemius fascia and detect its dynamic changes during ankle dorsiflexion. USWE can be used for biomechanical studies and intervention experiments concerning the deep fascia.

Effect of short latency stretch reflex on passive and active muscle stiffness in the soleus muscle in vivo

The purpose of the present study was to determine the effect of the short latency stretch reflex on passive and active muscle stiffness in the soleus muscle. Fourteen males participated in this study. Passive muscle stiffness was calculated from passive muscle force and fascicle length during passive lengthening at four different angular velocities (5, 50, 100, and 200degs-1). Active muscle stiffness was also calculated according to changes in muscle force and fascicle length during fast lengthening at five different angular velocities (100, 200, 300, 500, and 600degs-1) after submaximal isometric contraction (30% of maximal voluntary contraction). During the measurements of passive and active muscle stiffness, the amplitude of the short latency stretch reflex in the soleus muscle was measured. Change in passive torque and passive muscle stiffness significantly increased, whereas change in the fascicle length decreased, as angular velocity increased. At 100 and 200degs-1 (stretch reflex responses were not observed at 5 and 50degs-1), the amplitude of the short latency stretch reflex was highly correlated with passive muscle stiffness. Change in torque and active muscle stiffness were highest at 100degs-1 and decreased as angular velocity increased. At all angular velocities under active conditions, the amplitude of the short latency stretch reflex was not correlated with active muscle stiffness. These results suggest that the short latency stretch reflex affects passive muscle stiffness in the soleus muscle, but not active muscle stiffness.

Skeletal muscle titin IG domain refolding after passive lengthening

Titin is a structural protein which provides the majority of passive force in striated muscle sarcomeres. Evidence using titin fragments or in isolated I-band titin suggests that titin contributes to mechanical work in shortening muscle at physiologically relevant lengths by quick refolding of previously unfolded immunoglobulin domains. Quick refolding of Ig-domains has not been observed in sarcomeres, so this study tested whether rapid refolding of unfolded titin Ig-domains is observable in sarcomeres.

A new framework for analysis of three-dimensional shape and architecture of human skeletal muscles from in vivo imaging data.

A new framework is presented for comprehensive analysis of the three-dimensional shape and architecture of human skeletal muscles from magnetic resonance and diffusion tensor imaging data. The framework comprises three key features: 1) identification of points on the surface of and inside a muscle that have a correspondence to points on and inside another muscle, 2) reconstruction of average muscle shape and average muscle fiber orientations, and 3) utilization of data on between-muscle variation to visualize and make statistical inferences about changes or differences in muscle shape and architecture. The general use of the framework is demonstrated by its application to three case studies. Analysis of data obtained before and after 8 wk of strength training revealed there was little regional variation in hypertrophy of the vastus medialis and vastus lateralis and no systematic change in pennation angle. Analysis of passive muscle lengthening revealed heterogeneous changes in shape of the medial gastrocnemius and confirmed the ability of the methods to detect subtle changes in muscle fiber orientation. Analysis of the medial gastrocnemius of children with unilateral cerebral palsy showed that muscles in the more-affected limb were shorter, thinner, and less wide than muscles in the less-affected limb and had slightly more pennate muscle fibers in the central and proximal part of the muscle. Among other applications, the framework can be used to explore the mechanics of muscle contraction, investigate adaptations of muscle architecture, build anatomically realistic computational models of skeletal muscles, and compare muscle shape and architecture between species.NEW & NOTEWORTHY Muscle architecture is conventionally measured using simple scalar metrics such as muscle volume and average fascicle lengths. Here, a new framework is proposed for analysis of complex changes in three-dimensional architecture of whole human muscles from magnetic resonance and diffusion tensor imaging data. The general use of the framework is demonstrated through visualization, quantification, and statistical analysis of the effect of strength training, passive lengthening and cerebral palsy on three-dimensional muscle shape and architecture.

A 3D model of the soleus reveals effects of aponeuroses morphology and material properties on complex muscle fascicle behavior

The soleus is an important plantarflexor muscle with complex fascicle and connective tissue arrangement. In this study we created an image-based finite element model representing the 3D structure of the soleus muscle and its aponeurosis connective tissue, including distinct fascicle architecture of the posterior and anterior compartments. The model was used to simulate passive and active soleus lengthening during ankle motion to predict tissue displacements and fascicle architecture changes. Both the model’s initial architecture and changes incurred during passive lengthening were consistent with prior in vivo data from diffusion tensor imaging. Model predictions of active lengthening were consistent with axial plane muscle displacements that we measured in eight subjects’ lower legs using cine DENSE (Displacement Encoding with Stimulated Echoes) MRI during eccentric dorsiflexion. Regional strains were variable and nonuniform in the model, but average fascicle strains were similar between the compartments for both passive (anterior: 0.18 ± 0.06, posterior: 0.19 ± 0.05) and active (anterior: 0.12 ± 0.05, posterior: 0.13 ± 0.06) lengthening and were two- to three-times greater than muscle belly strain (0.06). We used additional model simulations to investigate the effects of aponeurosis material properties on muscle deformation, by independently varying the longitudinal or transverse stiffness of the posterior or anterior aponeurosis. Results of model variations elucidate how properties of soleus aponeuroses contribute to fascicle architecture changes. Greater longitudinal stiffness of posterior compared to anterior aponeurosis promoted more uniform spatial distribution of muscle tissue deformation. Reduced transverse stiffness in both aponeuroses resulted in larger differences between passive and active soleus lengthening.

Muscle architecture and passive lengthening properties of the gastrocnemius medialis and Achilles tendon in children who idiopathically toe-walk.

Children who idiopathically toe‐walk (ITW) habitually operate at greater plantarflexion angles and thus, at shorter muscle‐tendon unit (MTU) lengths than typically developing (TD) children. Therefore, it is often assumed that habitual use of the gastrocnemius muscle in this way will cause remodelling of the muscle‐tendon architecture compared to TD children. However, the gastrocnemius muscle architecture of children who ITW has never been measured. It is essential that we gain a better understanding of these muscle‐tendon properties, to ensure that appropriate clinical interventions can be provided for these children. Five children who ITW (age 8 ± 2 years) and 14 TD children (age 10 ± 2 years) participated in this study. Ultrasound was combined with isokinetic dynamometry and surface electromyography, to measure muscle architecture at common positions and passive lengthening properties of the gastrocnemius muscle and tendon across full range of motion. Regardless of which common condition groups were compared under, both the absolute and normalised to MTU muscle belly and fascicle lengths were always longer, and the Achilles tendon length was always shorter in children who ITW than TD children (p < 0.05; large effect sizes). The passive lengthening properties of the muscle and tendon were not different between groups (p > 0.05); however, passive joint stiffness was greater in children who ITW at maximum dorsiflexion (p = 0.001) and at a joint moment common to all participants (p = 0.029). Consequently, the findings of this pilot study indicate a remodelling of the relative MTU that does not support the concept that children who ITW commonly experience muscle shortening. Therefore, greater consideration of the muscle and tendon properties are required when prescribing clinical interventions that aim to lengthen the MTU, and treatments may be better targeted at the Achilles tendon in children who ITW.

Muscle Fascicles Exhibit Limited Passive Elongation Throughout the Rehabilitation of Achilles Tendon Rupture After Percutaneous Repair.

Achilles tendon rupture (ATR) results in long-term functional and structural deficits, characterized by reduced ankle mobility and plantarflexor muscle atrophy. However, it remains unclear how such functional impairments develop after surgical repair. While it is known that this injury negatively affects the tendon’s function, to date, limited work has focused on the short-term effect of ATR on the structure of the muscles in series. The aim of this study was to characterize changes in medial gastrocnemius architecture and its response to passive lengthening during the post-surgical rehabilitative period following ATR. Both injured and contralateral limbs from 10 subjects (1 female, BMI: 27.2 ± 3.9 kg/m2; age: 46 ± 10 years) with acute, unilateral ATR were assessed at 8, 12, and 16 weeks after percutaneous surgical repair. To characterize the component tissues of the muscle-tendon unit, resting medial gastrocnemius muscle thickness, fascicle length, and pennation angle were determined from ultrasound images with the ankle in both maximal plantarflexion and dorsiflexion. The ankle range of motion (ROM) was determined using motion capture; combined ultrasound and motion capture determined the relative displacement of the musculotendinous junction (MTJ) of the AT with the medial gastrocnemius. The ATR-injured gastrocnemius muscle consistently exhibited lower thickness, regardless of time point and ankle angle. Maximal ankle plantarflexion angles and corresponding fascicle lengths were lower on the injured ankle compared to the contralateral throughout rehabilitation. When normalized to the overall ankle ROM, both injured fascicles and MTJ displacement exhibited a comparably lower change in length when the ankle was passively rotated. These results indicate that when both ankles are passively exposed to the same ROM following ATR surgery, both ipsilateral Achilles tendon and gastrocnemius muscle fascicles exhibit limited lengthening compared to the contralateral MTU tissues. This appears to be consistent throughout the rehabilitation of gait, suggesting that current post-operative rehabilitative exercises do not appear to induce muscle adaptations in the affected MTU.

Corticospinal responses during passive shortening and lengthening of tibialis anterior and soleus in older compared to younger adults.

What is the central question of this study? Are there age-related differences in corticospinal responses whilst passively changing muscle length? What is the main finding and its importance? In contrast to young, older adults exhibited no modulation of corticospinal excitability in tibialis anterior during passive ankle movement. These data show impaired sensorimotor response in older adults during length changes of tibialis anterior, thus contributing to our understanding of age-related changes in sensorimotor control. Corticospinal responses have been shown to increase and decrease with passive muscle shortening and lengthening, respectively, as a result of changes in muscle spindle afferent feedback. The ageing sensory system is accompanied by a number of alterations that might influence the processing and integration of sensory information. Consequently, corticospinal excitability might be modulated differently whilst changing muscle length. In 10 older adults (66±4years), corticospinal responses (MEP/Mmax ) were evoked in a static position, and during passive shortening and lengthening of soleus (SOL) and tibialis anterior (TA), and these data were compared to the re-analysed data pool of 18 younger adults (25±4years) published previously. Resting motor threshold was greater in SOL compared to TA (P<0.001), but did not differ between young and older (P=0.405). No differences were observed in MEP/Mmax between the static position, passive shortening or lengthening in SOL (young: all 0.02±0.01; older: 0.05±0.04, 0.03±0.02 and 0.04±0.01, respectively; P=0.298), and responses were not dependent on age (P=0.090). Conversely, corticospinal responses in TA were modulated differently between the age groups (P=0.002), with greater MEP/Mmax during passive shortening (0.22±0.12) compared to passive lengthening (0.13±0.10) and static position (0.10±0.05) in young (P<0.001), but unchanged in older adults (0.19±0.11, 0.22±0.11 and 0.18±0.07, respectively; P≥0.867). The present experiment shows that length-dependent changes in corticospinal excitability in TA of the young are not evident in older adults. This suggests impaired sensorimotor response during muscle length changes in older age that might only be present in ankle flexors, but not extensors.

Mechanical factors tune the sensitivity of mdx muscle to eccentric strength loss and its protection by antioxidant and calcium modulators

BackgroundDystrophin deficiency sensitizes skeletal muscle of mice to eccentric contraction (ECC)-induced strength loss. ECC protocols distinguish dystrophin-deficient from healthy, wild type muscle, and test the efficacy of therapeutics for Duchenne muscular dystrophy (DMD). However, given the large lab-to-lab variability in ECC-induced strength loss of dystrophin-deficient mouse skeletal muscle (10–95%), mechanical factors of the contraction likely impact the degree of loss. Therefore, the purpose of this study was to evaluate the extent to which mechanical variables impact sensitivity of dystrophin-deficient mouse skeletal muscle to ECC.MethodsWe completed ex vivo and in vivo muscle preparations of the dystrophin-deficient mdx mouse and designed ECC protocols within physiological ranges of contractile parameters (length change, velocity, contraction duration, and stimulation frequencies). To determine whether these contractile parameters affected known factors associated with ECC-induced strength loss, we measured sarcolemmal damage after ECC as well as strength loss in the presence of the antioxidant N-acetylcysteine (NAC) and small molecule calcium modulators that increase SERCA activity (DS-11966966 and CDN1163) or lower calcium leak from the ryanodine receptor (Chloroxine and Myricetin).ResultsThe magnitude of length change, work, and stimulation duration ex vivo and in vivo of an ECC were the most important determinants of strength loss in mdx muscle. Passive lengthening and submaximal stimulations did not induce strength loss. We further showed that sarcolemmal permeability was associated with muscle length change, but it only accounted for a minimal fraction (21%) of the total strength loss (70%). The magnitude of length change also significantly influenced the degree to which NAC and small molecule calcium modulators protected against ECC-induced strength loss.ConclusionsThese results indicate that ECC-induced strength loss of mdx skeletal muscle is dependent on the mechanical properties of the contraction and that mdx muscle is insensitive to ECC at submaximal stimulation frequencies. Rigorous design of ECC protocols is critical for effective use of strength loss as a readout in evaluating potential therapeutics for muscular dystrophy.

What causes increased passive stiffness of plantarflexor muscle-tendon unit in children with spastic cerebral palsy?

The term 'stiffness' is commonly used in the literature to refer to various components of 'hyperresistance' by which spastic muscles oppose to their passive lengthening, especially in children with cerebral palsy (CP). Originally, stiffness consists of mechanical resistance to passive movement in the absence of any muscle activation. Increased muscle stiffness in CP therefore refers to alterations to the mechanical properties of the tissue. It is closely linked to muscle shortening, yet the two phenomena are not equivalent. Both increased stiffness and shortening are present early in childhood in the plantarflexor muscles of children with spastic CP. This narrative review provides a comprehensive overview of the literature on passive stiffness of the plantarflexor muscles measured at the joint, muscles, fascicles, and fiber level in children with CP. Articles were searched through the Pub'Med database using the keywords "cerebral palsy" AND "stiffness". The ambiguous use of the term 'stiffness' has been supported by discrepancies in available results, influenced by heterogeneity in materials, methodologies and characteristics of the participants among studies. Increased stiffness at the joint and muscle belly level may be explained by altered structural properties at the microscopic level. This thorough investigation of the literature suggests that the pathophysiology and the time course of the development of stiffness and contracture remain to be elucidated. A consideration of both morphological and mechanical measurements in children with CP is important when describing the alterations in their plantarflexors.

The association of passive medial gastrocnemius muscle tendon unit lengthening behaviour with ankle gait pattern in children with cerebral palsy

The association of passive medial gastrocnemius muscle tendon unit lengthening behaviour with ankle gait pattern in children with cerebral palsy

Corticospinal excitability of tibialis anterior and soleus differs during passive ankle movement

The purpose of this study was to assess corticospinal excitability of soleus (SOL) and tibialis anterior (TA) at a segmental level during passive ankle movement. Four experimental components were performed to assess the effects of passive ankle movement and muscle length on corticospinal excitability (MEP/Mmax) at different muscle lengths, subcortical excitability at the level of lumbar spinal segments (LEP/Mmax), intracortical inhibition (SICI) and facilitation (ICF), and H-reflex in SOL and TA. In addition, the degree of fascicle length changes between SOL and TA was assessed in a subpopulation during passive ankle movement. Fascicles shortened and lengthened with joint movement during passive shortening and lengthening of SOL and TA to a similar degree (p < 0.001). Resting motor threshold was greater in SOL compared to TA (p ≤ 0.014). MEP/Mmax was facilitated in TA during passive shortening relative to the static position (p ≤ 0.023) and passive lengthening (p ≤ 0.001), but remained similar during passive ankle movement in SOL (p ≥ 0.497), regardless of muscle length at the point of stimulus (p = 0.922). LEP/Mmax (SOL: p = 0.075, TA: p = 0.071), SICI (SOL: p = 0.427, TA: p = 0.540), and ICF (SOL: p = 0.177, TA: p = 0.777) remained similar during passive ankle movement. H-reflex was not different across conditions in TA (p = 0.258), but was reduced during passive lengthening compared to shortening in SOL (p = 0.048). These results suggest a differential modulation of corticospinal excitability between plantar and dorsiflexors during passive movement. The corticospinal behaviour observed might be mediated by an increase in corticospinal drive as a result of reduced afferent input during muscle shortening and appears to be flexor-biased.

Repeated anodal trans-spinal direct current stimulation results in long-term reduction of spasticity in mice with spinal cord injury.

Spasticity is a disorder of muscle tone that is associated with lesions of the motor system. This condition involves an overactive spinal reflex loop that resists the passive lengthening of muscles. Previously, we established that application of anodal trans-spinal direct current stimulation (a-tsDCS) for short periods of time to anaesthetized mice sustaining a spinal cord injury leads to an instantaneous reduction of spasticity. However, the long-term effects of repeated a-tsDCS and its mechanism of action remained unknown. In the present study, a-tsDCS was performed for 7 days and this was found to cause long-term reduction in spasticity, increased rate-dependent depression in spinal reflexes, and improved ground and skill locomotion. Pharmacological, molecular and cellular evidence further suggest that a novel mechanism involving Na-K-Cl cotransporter isoform 1 mediates the observed long-term effects of repeated a-tsDCS. Spasticity can cause pain, fatigue and sleep disturbances; restrict daily activities such as walking, sitting and bathing; and complicate rehabilitation efforts. Thus, spasticity negatively influences an individual's quality of life and novel therapeutic interventions are needed. We previously demonstrated in anaesthetized mice that a short period of trans-spinal subthreshold direct current stimulation (tsDCS) reduces spasticity. In the present study, the long-term effects of repeated tsDCS to attenuate abnormal muscle tone in awake female mice with spinal cord injuries were investigated. A motorized system was used to test velocity-dependent ankle resistance and associated electromyographical activity. Analysis of ground and skill locomotion was also performed, with electrophysiological, molecular and cellular studies being conducted to reveal a potential underlying mechanism of action. A 4week reduction in spasticity was associated with an increase in rate-dependent depression of spinal reflexes, and ground and skill locomotion were improved following 7days of anodal-tsDCS (a-tsDCS). Secondary molecular, cellular and pharmacological experiments further demonstrated that the expression of K-Cl co-transporter isoform 2 (KCC2) was not changed in animals with spasticity. However, Na-K-Cl cotransporter isoform 1 (NKCC1) was significantly up-regulated in mice that exhibited spasticity. When mice were treated with a-tsDCS, down regulation of NKCC1 was detected, and this level did not significantly differ from that in the non-injured control mice. Thus, long lasting reduction of spasticity by a-tsDCS via downregulation of NKCC1 may constitute a novel therapy for spasticity following spinal cord injury.

Exercising Bioengineered Skeletal Muscle In Vitro: Biopsy to Bioreactor.

The bioengineering of skeletal muscle tissue in-vitro has enabled researchers to more closely mimic the in-vivo skeletal muscle niche. The three-dimensional (3-D) structure of the tissue engineered systems employed to date enable the generation of highly aligned and differentiated myofibers within a representative biological matrix. The use of electrical stimulation to model concentric contraction, via innervation of the myofibers, and the use of mechanical loading to model passive lengthening or stretch has begun to provide a manipulable environment to investigate the cellular and molecular responses following exercise mimicking stimuli in-vitro. Currentlyavailable bioreactor systems allow either electrical stimulation or mechanical loading to be utilized at any given time. In the present manuscript, we describe in detail the methodological procedures to create 3-D bioengineered skeletal muscle using both cell lines and/or primary human muscle derived cells from a tissue biopsy, through to modeling exercising stimuli using a bioreactor that can provide both electrical stimulation and mechanical loading simultaneously within the samein-vitro system.

Transient Increase in Cortical Excitability Following Static Stretching of Plantar Flexor Muscles.

Spinal excitability in humans is inhibited by both passively holding a static position with the muscle lengthened (static stretching) and by a single non-active lengthening movement. However, whilst immediately after a passive lengthening movement the inhibition persists for several seconds, there seem to be an immediate recovery following static stretching. This result is counter intuitive and could be attributed to methodological procedures. Indeed, differently to what has been done until now, in order to study whether static stretching has a transient effect on the neuromuscular pathway, the procedure should be repeated many times and measurements collected at different time points after stretching. In the present study we repeated 60 times 30 s static stretching of ankle plantar flexors and measured tap reflex (T-reflex), Hoffman reflex (H-reflex), and motor evoked potentials (MEPs) from the Soleus muscle at several time points, starting from immediately after until 30 s following the procedure. T-reflex was strongly inhibited (range 31–91%, p = 0.005) and the inhibition persisted for 30 s showing a slow recovery (r = 0.541, p = 0.037). H-reflex was not affected by the procedure. Stretching increased the size of the MEPs (p < 0.0001), differences at times 0 and 2 s after stretching (p = 0.015 and p = 0.047, respectively). These results confirm that static stretching reduces muscle spindle sensitivity. Moreover it is suggested that post-activation depression of Ia afferents, which is commonly considered the cause of H-reflex depression during both dorsiflexion and static stretching, vanished immediately following stretching or is counteracted by an increased corticospinal excitability.

Apophyseal avulsion fractures of the pelvis. A review.

Background and aim of the work: Fractures of the pelvis classically occur in adolescent during sports activities with forceful and repetitive contractions or passive lengthening acting on not yet ossified growth plates. Their misdiagnosis lead to disability, chronic pain and decrease of performances. Evidence based treatment guidelines do not exist; aim of this paper is to point out clinical outcomes, return to sport rates and complications of surgical and conservative approach. Methods: A systematic search based on MEDLINE database was performed in August 2017 to identify all published articles from 2010 to 2017 reporting outcomes, return to sport and complications rates after surgical and non-operative treatment of avulsion fractures of the pelvis. Results: Mean age was 14,5 years with anterior inferior iliac spine avulsion representing the most common injury (46%), followed by anterior superior iliac spine avulsion (32%), ischial tuberosity avulsion (12%) and iliac crest avulsion (11%). Rates of excellent outcome and return to sports at pre-injury levels were higher after surgical treatment; surgery has a higher risk of heterotopic ossification (9%) compared to conservative treatment (1,8%), whereas the risk of non-unions is lower (0% versus 2,5%). Conclusions: Surgery is preferred for major dislocations and fragment sizes, providing a faster return to pre-injury level of activity, decreasing the risk of pseudoarthrosis. Conservative treatment is advisable for minimally displaced fractures when a rapid recovery is not required; patient and his family should be informed on the risk of non-unions and the eventuality of a delayed surgical approach. (www.actabiomedica.it)