key: cord-0029717-pyamofjk
authors: Sweeney, H. Lee; Masiero, Stefano; Carraro, Ugo
title: The 2022 On-site Padua Days on Muscle and Mobility Medicine hosts the University of Florida Institute of Myology and the Wellstone Center, March 30 - April 3, 2022 at the University of Padua and Thermae of Euganean Hills, Padua, Italy: The collection of abstracts
date: 2022-03-10
journal: Eur J Transl Myol
DOI: 10.4081/ejtm.2022.10440
sha: ad04069287e3bf55085cc4e67fb0a7c91255e83c
doc_id: 29717
cord_uid: pyamofjk

In the autumn of 2021, the 2022 Padua Days of Muscle and Mobility Medicine (PDM3) was planned to be held from March 30 to April 2, 2022. Despite the fact that Coronavirus COVID-19 outbreak continued to impose restrictions all over the world, the program was planned with Scientific Sessions to occur over three full days at either the University of Padua Aula Magna or the Hotel Petrarca on the Thermae of Euganean Hills (Padua), Italy. During the winter the epidemic worsened, but waned by early February 2022, allowing confirmation of the planned On-site Meeting. The success of submission of abstracts (over 100 abstracts, i. e., three times the previous years) is attributable to the fact that the 2022 On-site PDM3 is a combined meeting, hosting the spring Meeting of the Myology Institute and Wellstone Center of the University of Florida, USA. The first three days will include a large series of presentations of the University of Florida scientists and clinicians and of their Invited Speakers from Canada, France, Switzerland Italy and the U.K. European researchers and clinicians from France, Germany, Iceland, Ireland, Italy, Slovenia, Russia and UK fill the program of last two days. To provide slot times to young Speakers, the 2022 On-site PDM3 was extended to April 3, 2022. The Collection of Abstracts is e-published in the 32(1) 2022 Issue of the European Journal of Translational Myology (EJTM), together with the detailed Program organized in the Aula Magna of the University of Padua (March 30) and for March 31 - April 3, 2022 at the Conference Halls of Hotel Petrarca of the Thermae of Euganean Hills (Padua), Italy. The Program ends late on Sunday April 3, 2022 with an invitation to join the 2023 PDM3, March 27 – 31 at the Thermae of Euganean Hills (Padua), Italy. You are invited to join and to submit your Communications to the European Journal of Translational Myology and to a Special Issue of the Journal diagnostics, MDPI, Basel. Both journals will provide 50% discount to the first 15 accepted typescripts.

The ubiquitin proteasome system (UPS) is a major player of skeletal muscle wasting, a common characteristic of many diseases (cancer, sepsis, heart failure, kidney diseases, etc.) that negatively impacts treatment and life prognosis. In the early 2000, the notion of "atrogenes" (atrophy-related genes) was proposed for genes that were systematically up regulated during catabolic situations in rodents, which included 2 founding members, the E3 ubiquitin ligases MAFbx/Atrogin-1 and MuRF1/TRIM63. 1 Interestingly, MuRF1 is so far the only E3 ligase known for targeting several sarcomeric proteins (a-actin, MYHC, Troponin I, telethonin) and we recently identified the E2 ubiquitin conjugating enzymes that bring the catalytic activity for MuRF1-dependent protein degradation. 2, 3 Besides rodent models, we and others demonstrated that MAFbx and MuRF1 are also gold standard atrogenes in human pathologies and we recently found that numerous proteins may be part of a muscle atrophy program in chronic kidney disease and lung cancer patients. 4 In addition, we have identified blood markers that reflect skeletal muscle loss in patients suffering from several pathologies. Maintaining the integrity of organelles and proteins despite the cellular stresses that arise from environmental pressure is essential for life. The time-dependent accumulation of cellular damage is widely considered to be the general cause of cellular degeneration. Decline of the protein quality control, called the loss of proteostasis, contributes to impairment of macromolecule function and is considered among the primary hallmark of aging and disease onset. All cells take advantage of an array of mechanisms to preserve the stability and functionality of their proteins or to remove them when they are irreversibly damaged. The two the most important cellular housekeeping and pro-survival pathway are autophagy and ubiquitin proteasome (UPS) whose main action is to remove damaged proteins/organelles and generate molecules that sustain cellular core metabolism. Autophagy, like the ubiquitin-proteasome system, strictly regulates diverse cellular functions through its selectivity. I will describe the mutual regulation between the UPS and autophagy and their involvement in protein turnover and disease onset in post mitotic tissues.

***** Unfortunately, there are no currently available medical treatments for cancer-induced muscle pathology which is due, in part, to an incomplete understanding of the molecular mechanisms which are causative in the condition. The E3 ubiquitin ligase MuRF1 is required for muscle atrophy in several pathophysiological conditions which associate with muscle atrophy. Here we tested the requirement of MuRF1 for cancer-induced muscle atrophy using MuRF1 KO (MuRF1-/-) mice, or wild type (WT) controls, and an orthotopic pancreatic cancer (KPC) model. WT mice reached experimental endpoint 14 to 17 days following KPC cell injection, as defined by significant body weight loss (-12%) and deterioration of body condition score, with an average tumor mass of 1.9 ± 0.5 g. At this same time point, MuRF1-/-mice were weight stable (-0.6%), and showed ~84% lower tumor mass (0.3 ± 0.1 g) compared to WT mice. When allowing tumor growth to progress in MuRF1-/-mice, it took 21-36 days to reach a comparable tumor mass to that of WT mice at days 14-17. Moreover, when WT and MuRF1-/mice presenting matching tumor sizes were compared, MuRF1-/-mice showed preserved muscle mass, with no muscle wasting observed in the triceps surae (-0.1% in MuRF1-/-vs. -9% in WT mice) and attenuated wasting observed in tibialis anterior (-5% in MuRF1-/-vs. -18% in WT mice). In addition to preserving muscle mass, MuRF1 deletion also protected against cancer-induced muscle weakness. Indeed, the soleus and diaphragm of WT mice showed a 32% and 46% reduction in specific tetanic force, respectively, whereas no such muscle weakness was observed in MuRF1-/-mice (+20% and +3% in soleus and diaphragm respectively). Skeletal muscle proteome and ubiquitinome analyses were conducted to examine the mechanisms through which MuRF1 deletion conferred its protection against cancerassociated muscle wasting and dysfunction. These analyses revealed that KPC-tumor-burden induces, in a MuRF1-independent manner, an early and sustained increase in the abundance of proteins involved in the acute phase response as well as in extracellular matrix remodeling, while proteins involved in protein translation and oxidative phosphorylation showed a MuRF1-dependent reduction in their abundance at time points reflective of cachexia initiation and progression. Identification of proteins ubiquitinated in a MuRF1dependent manner further revealed an early and sustained enrichment in proteins related to muscle contraction, proteasome complex and glucose homeostasis. In conclusion, our work suggests that MuRF1 is required for the normal muscle wasting and weakness that occurs in KPC tumor bearing mice, and is also required for normal tumor growth. This suggests that MuRF1-dependent muscle protein breakdown may be required to supply the tumor with nutrients needed for its growth. Sarcopenia is defined as the loss of muscle mass and function and is considered a major contributor to loss of life quality at high age. The mammalian target of rapamycin complex 1 (mTORC1) is well known to regulate cell size by controlling cap-dependent protein translation in response to growth factors and nutrients. In skeletal muscle, the mTORC1 inhibitor rapamycin prevents load-induced hypertrophy. In contrast to expectations, mTORC1 signaling is high in sarcopenic muscle, its constant activation causes atrophy in most muscles and prolonged treatment with rapamycin (i.e. inhibition of mTORC1) prolongs lifespan in many species. Based on these observations, we have addressed the role of PKB/Akt-mTORC1 signaling in age-related sarcopenia. We find that i) mice depleted of the mTORC1 inhibitor TSC1 in skeletal muscle show a phenotype reminiscent of sarcopenia and ii) inhibition of mTORC1 by rapamycin has an overly anti-sarcopenic effect. Molecular analysis also shows a strong overlap in the gene signatures of aging and mTORC1 activation. We also show that caloric restriction (CR), the best-studied anti-aging intervention thought to act via inhibition of the nutrient-sensing mTORC1, has an anti-sarcopenia effect.

Interestingly, molecular analysis reveals that the CR effect is largely distinct from the effect of rapamycinmediated mTORC1 inhibition. Consistent with this, the combination of CR and rapamycin has additive effects on the prevention of sarcopenia. Finally, we also provide evidence that hyperactivation of PKB/Akt is detrimental to muscle function in skeletal muscle with high mTORC1 activity. In summary, our data indicate that great caution should be exercised with intervention strategies designed to increase muscle size by interfering with the pathways that control the balance between protein synthesis and protein degradation Purpose of this study was to assess the effect of a course of homeopathic preparations using acupuncture points on the severity of Cancer cachexia is a multifactorial condition characterized by skeletal muscle loss that impairs longevity and quality of life of the vast majority of cancer patients. 1, 2 However, the ability to develop therapeutic strategies to counter cachexia is impeded by the limited understanding of the underlying mechanisms of cachexia in human cancer patients. The purpose of this study was therefore to identify the proteomic signature of skeletal muscle obtained from cachectic pancreatic ductal adenocarcinoma (PDAC) patients, who exhibit one of the highest rates of cachexia. 3, 4 Muscle biopsies of the rectus abdominis were obtained from cachectic PDAC patients (n=8) undergoing tumor resection surgery as well as age and sex-matched non-cancer controls (n=6). Total proteome profiling of skeletal muscle protein isolate was performed using 10-plex tandem mass tag labeling and liquid chromatography tandem mass spectrometry (LC-MS/MS). MS/MS spectra were evaluated using SEQUEST and differentially expressed proteins were identified through the UniProt Homo Sapiens database [adjusted P value (FDR) ≤0.01; -1.25≥fold change≥1.25].

The total number of identified peptides and proteins are shown in Figure 1 , which further shows that 383 proteins were identified to be differentially expressed in the skeletal muscle of PDAC patients compared to controls. To identify altered biological processes within the skeletal muscle of cachectic PDAC patients, differentially expressed proteins (DEP) were analyzed using several bioinformatic platforms. The results of these bioinformatic analyses will be presented, along with immunohistochemical analyses to further support changes in select proteins of interest.

Muscle atrophy is an inevitable consequence of chronic disuse. Until recently, in non-pathological conditions, the main cause of muscle wasting induced by inactivity was considered to be the lack of mechanical loading, resulting in a depression of protein synthesis and an increase in protein breakdown. However, there is now increasing evidence that neurodegenerative processes, triggered by inactivity, contribute to the loss of muscle mass and function. Indeed, instability of the neuromuscular junction (NMJ) and onset of denervation/reinnervation processes have been recently found after short-term and long-term inactivity in humans caused by bed rest or (Fig. 1) . These changes are also accompanied by instability of the NMJ, as shown either by increased serum levels of c-terminal agrin fragment (CAF) after 10day bed rest and after 10-day ULLS, or by a decreased expression of Homer proteins in muscle biopsies, after 60-day bed rest (Salanova et al 2011). These findings of NMJ instability and partial fibre denervation were associated with an impairment of sarcoplasmic reticulum (SR) calcium handling, since a significant depression in the caffeine-induced release of Ca2+ by the SR was found after 10-day bed rest (Monti et al. 2021) , suggesting an impairment of excitation-contraction coupling. These findings support the view that the early onset of NMJ instability and impairment in SR function are eligible mechanisms contributing to the greater decline in muscle force than in muscle size observed in these bed rest studies and in most models of muscle disuse. Acknowledgements: Funding by ASI, MARS- Interleukin-6 (IL-6) is a soluble mediator with pleiotropic actions on different tissues and organs. The multifaceted nature of IL-6 action can be ascribed to the multi-level regulation of the different signaling pathways that it is able to induce. Although IL-6 is produced by several cell types and it can signal through different receptor complexes, inducing a variety of intracellular pathways, its expression and signaltransduction is strictly controlled (Figure 1 ). IL-6 expression is regulated at the transcriptional and posttranscriptional level, while the induction of the signaling cascade is negatively regulated by systems of internalization/degradation of receptor subunits and by the action of negative feedback molecules. Thus, acting as a warning molecule, the abundance and persistence of IL-6 need a fine regulation in order to limit the powerful action of the cytokine at the physiologic stimuli. IL-6 overexpression is able to exacerbate the dystrophic muscle phenotype, fostering pathologic changes in skeletal muscle and closely approximating the disease progression of DMD patients, 1,2 whereas short inhibition of IL-6 activities, by a neutralizing antibody directed against IL6R, is beneficial to dystrophic muscle decreasing inflammation−induced myonecrosis, favoring the maintenance of muscle function, and preventing exercise-induced fiber damage and loss of muscle strength. 3 In addition, overexpression of circulating IL-6 enhances the generation and accumulation of free radicals, influencing the local redox homeostasis, inducing the establishment of prooxidative conditions in skeletal muscle tissue. 4 Key Words: IL-6; muscle homeostasis; muscle regeneration. (DOX) is a highly effective chemotherapeutic agent used to treat a wide variety of cancers. However, its clinical use is associated with respiratory muscle weakness, resulting in fatigue, dyspnea and exercise intolerance. 1 While no therapeutic countermeasures exist to prevent the development of DOX-induced respiratory dysfunction, previous preclinical work has demonstrated that acute endurance exercise preconditioning prior to DOX administration was sufficient to preserve diaphragm muscle force production, maintain fiber cross-sectional area and prevent ventilatory deficits during a hypercapnic -23 -challenge. These protective effects were associated with the exercise-induced upregulation of the mitochondrialocalized ATP-binding cassette transporters in the diaphragm, including ABCB6, which is hypothesized to play a role in maintaining mitochondrial redox balance and heme homeostasis. 2 To determine if ABCB6 overexpression, independent of exercise, is sufficient to prevent DOX-induced respiratory dysfunction, female Sprague-Dawley rats were randomly assigned to receive either rAAV9-MHCK7-rABCB6 to overexpress ABCB6 in the diaphragm or saline. These groups were further divided to receive saline or DOX (20mg/kg i.p.) treatment. Performance of an exercise tolerance test forty-eight hours following treatment revealed significant reductions in run time to fatigue in both DOXtreated groups. However, ventilatory function in response to hypercapnia was only reduced in the saline-saline rats compared to the saline-DOX rats. Additionally, ABCB6 overexpression attenuated the DOX-induced reduction in diaphragm muscle force production. These findings support a role for ABCB6 in the preservation of respiratory function following acute exposure to DOX. Low extremity arterial disease (LEAD) is the most common form of peripheral vascular disease and is often a direct consequence of the atherosclerotic process. Characterised by chronic decreases in lower limb blood flow, this progressive disease results in symptoms ranging from pain, muscle weakness, to severe exercise intolerance. Several animal models have been used to study the angiogenesis and muscle remodelling process Skeletal muscle function is regulated by intracellular Ca 2+ levels. Two main mechanisms control movements of Ca 2+ ions from intracellular stores (i.e. the sarcoplasmic reticulum, SR) and from extracellular space: i) excitation-contraction (EC) coupling; and ii) store-operated Ca 2+ entry (SOCE). SOCE allows recovery of extracellular Ca2+ during prolonged muscle activity, when the SR undergoes depletion. We recently discovered that prolonged exercise leads to formation of Calcium Entry Units (CEUs),1 intracellular junctions located at the I band that are formed by two distinct elements: SR-stacks and transverse tubules (TTs)1,2. Assembly of CEUs during exercise promotes the interaction between STIM1 and Orai1, the two main proteins that mediate SOCE, and increases muscle resistance to fatigue in presence of extracellular Ca 2+ . The molecular mechanisms underlying the exercisedependent remodeling of SR and TT leading to CEU assembly remain to be fully elucidated. 3 Here, we first verified whether CEUs can assemble ex-vivo (in absence of blood supply and innervation) subjecting excised EDL muscles from mice to an ex-vivo incremental fatigue protocol (80Hz tetanus stimulation lasting 45 minutes): the data collected demonstrate that CEUs can assemble ex-vivo in isolated EDL muscles. We then evaluated if intracellular parameters that are affected by exercise, such as temperature and pH, may influence the assembly of CEUs. We found that higher temperature (36°C vs. 25°C) and lower pH (7.2 vs. 7.4) promotes formation of CEUs increasing the % of fibers containing SR stacks, the n. of SR stacks/area, and also the elongation of TTs at the I band. Importantly, increased assembly of CEUs at higher temperature (36 o C) or at lower pH (7.2) Skeletal muscle is a post-mitotic tissue, yet has the ability to regenerate through the activation of muscle satellite cells and the coordinated actions of multiple growth factors. A key player in the regeneration process is Insulinlike Growth Factor-I (IGF-I). 1 IGF-I helps to resolve damage by promotion of satellite cell proliferation and differentiation, suppression of proinflammatory cytokines, and fiber formation. A longstanding goal in our laboratory has been to optimize IGF-I forms to improve its actions in skeletal muscle. While the most well characterized form of IGF-I is the small mature 7kD protein, complexity of alternative splicing and posttranslational modification leads to several additional forms of this growth factor. These larger forms have been shown to have more efficient storage in muscle tissue as well as enhanced receptor activation, 2,3 and raise the possibility that improvements in therapeutic potential could occur through the use of a different IGF-I form. To understand the necessity of glycosylated and mature IGF-I forms, gene editing was employed to remove glycosylation sites in the C-terminal portion of IGF-IA, and to eliminate the ability for cleavage of the C-terminal extensions of all IGF-I forms ( Figure 1 ). 4 Mice were viable, with normal force generation. To determine the effects on regeneration after acute injury, cardiotoxin injections were performed in male and female mice from all genotypes, and monitored for changes in fiber size. In both 2ND and 3RA genotypes, there was an enhanced recovery of fiber size following injury. This suggests that the pro-IGF-I forms, common to both genotypes may contribute to accelerating the resolution of damage in muscle. Future studies will take advantage of these forms to develop more effective therapeutics for muscle. Impaired mitochondrial function and abnormal muscle bioenergetics are evident in several diseases and aging and have been associated with loss of muscle contractile function. However, a direct link between metabolic dysfunction and loss of muscle force or power has not been established. Mitochondrial respiration impairments lead to accumulation of metabolites of the tricarboxylic acid cycle that are upstream of the electron transport system such as succinate and succinyl-derivatives. 1 High fat diet, diabetes, hypoxia/ischemia, cancer, and even exercise disrupt the metabolism succinate and succinylderivatives resulting in several fold increase in succinate. [1] [2] [3] [4] [5] We have found that postmenopausal heart failure with preserved ejection fraction (HFpEF) causes muscle mitochondrial dysfunction and weakness, 6 which are accompanied by elevated plasma [succinate] that reflects increased cellular concentration and export from the most metabolically active tissues: adipose, liver, and skeletal muscle. This presentation will focus on preliminary findings showing that: 1) HFpEF disrupts the abundance of proteins involved in mitochondrial succinate transport and lysine succinylation, resulting in increased myofibrillar protein succinylation; 2) acute exposure of intact skeletal muscle and permeabilized single fiber to succinate, which causes succinylation of myofibrillar proteins, impairs isometric and isotonic contractile function; and 3) the effects of succinate in vitro are independent of ROS from mitochondrial complex II. Overall, our findings suggest that succinate and lysine succinylation are a novel link between metabolic and contractile dysfunction in health and disease.

Key Words: Heart failure; succinate; weakness; peak power.

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Higher abundance of deletions and strand break damage within specific mitochondrial ETC genes are associated with functional performance in older adults.

Sunil K. Saini The goal of this study was to investigate if higher abundance of deletions and strand break damage within specific Mt ETC genes are associated with functional performance in older adults. Healthy older adults (65.2 ± 6.7 years; n = 25) were enrolled in this study (1) . The participants completed a 6-minute walk test with an average 6-minute walking distance of 420 (± 149) meters and a usual-paced walking speed over 4 meters of 0.86 (± 0.28) meters per second. We quantified in muscle-biopsy specimen the abundance of mtDNA deletion and SB damage and associated these with the recorded walking speed. Specifically, mtDNA regions were selected based on previous findings from our group as potential hotspots for damage (2) . We determined (a) the abundance of the 4977 bp deletion, a common deletion present in human mtDNA, which results in loss of a mtDNA fragment that includes genes coding for ATPase6, ATPase8, cytochrome oxidase III, NADH dehydrogenase subunit 3 (ND3), ND4, ND4 subunit L (ND4L), and ND5; and (b) the abundance of mtDNA strand break damage within specific mtDNA regions that code for subunits of Complex I (ND1, ND2, ND4, ND5), Complex III (Cytochrome B6), Complex IV (Cytochrome Oxidase subunit II), and Complex V (ATPase subunit 6/8), as well as SB in the regulatory non-coding D-loop region. Please note that the highly sensitive mtDNA strand break method spans through regions coding for several ETC genes. We plotted the robust correlations of mitochondrial SBs in specific mtDNA regions with walking performance in 25 older adults (Fig). For example, a greater abundance of density of mtDNA SB damage (SB damage normalized to mtDNA copy number) within NADH dehydrogenase subunit 1/2 (ND1/2) and 4/5 (ND4/5) was strongly associated with poor walking performance (r = -0.59 and r = -0.70). Our data showed the powerful association and sensitivity of mtDNA SB in specific Mt ETC regions with walking performance. Conclusio n: This set of data indicates that mtDNA mutations in ETC components (strand breaks to ND1/2 and ND4/5) are strongly related to slower walking speed in older adults. inhibition of proliferation and blunted the onset of cell death in human myoblasts ( Figure 1 ). Functionally, DUX4 forces the nuclear translocation of β-CATENIN and disrupting canonical WNT signalling. However, concomitant overexpression of DUX4c restores β-CATENIN cytoplasmic/nuclear shuttling and the downstream transcriptional program (Figure 1 ). We identified a subset of genes involved in the β-CATENIN pathway that are differentially regulated between DUX4 and DUX4c and can separate FSHD from healthy muscle biopsies based on the expression pattern. Constitutive expression of DUX4c robustly suppresses expression of DUX4 target genes confirming molecular dueling. In line, FSHD myoblasts display upregulation of DUX4 and downregulation of DUX4c compared to aged-matched unaffected myoblast ( Figure 2 ). Finally, blockade of WNT/β-CATENIN pathway rescue proliferation defect of FSHD myoblasts ( Figure 2 ). Together, our study demonstrates that DUX4 alters cell viability via β-CATENIN activation and that DUX4c can counteract aspects of DUX4-mediated toxicity in human muscle cells, informing future interventions to ameliorate FSHD pathology. The p97 complex participates in the degradation of muscle proteins during atrophy upon fasting or denervation interacting with different adaptors. 1 We investigated whether and how it might also be involved in muscle wasting in cancer, where loss of appetite occurs, or amyotrophic lateral sclerosis (ALS), where motoneuron death causes muscle denervation and fatal paralysis. 2 The mRNA levels of p97 were induced in tibialis anterior (TA) of three distinct cancer cachectic mouse models but not in the non-cachectic 4T1 tumor. Similarly, p97 was high both in mRNA and protein in muscles from SOD1G93A mice. Electroporation of a shRNA for murine p97 into muscle reduced the fiber atrophy caused by colon adenocarcinoma C26 and ALS. When we interrogated a microarray we had previously generated for the expression of p97 adaptors, we found few of them induced in cachectic TA from C26-mice. By qPCR, we validated their inductions in TA of cachectic and ALS models and selected Nploc4 as the one most induced. Electroporation of a Crispr/Cas9 vector against Nploc4 into muscle reduced the fiber atrophy caused by C26 and ALS. Since Disulfiram (DSF) uncouples p97 from Nploc4, 3 we treated atrophying myotubes with DSF, and found accumulated polyubiquitinated proteins and reduced degradation of long-lived proteins. DSF halves Nploc4 in the soluble muscle fraction and given to C26-mice limited the body and muscle weight loss, with no effect on tumor. The p97/Nploc4 complex plays a crucial role in muscle atrophy during these disorders and disrupting this complex might serve as a novel drug strategy. A number of different microdystrophin constructs are at various stages of clinical development. Early data on microdystrophin protein production in the biopsies of individuals treated in the higher dose categories are encouraging, but as the programs proceed, a number of issues that the field needs to address are becoming more evident:

• Expectations. The degree of functional improvement following AAV gene therapy has attracted a lot of attention as if this was an essential aspect of the outcome, but in reality the protection of transduced muscle from progressive damage is a more relevant outcome. • Durability. As the transduced protein is a Becker-like microdystrophin, complete protection from further damage may be challenging, although the levels at which this AAV transgene is produced is significantly higher than those that BMD patients typically produce. • Efficacy in all tissues. Many of these microdystrophins lack domains that, in BMD patients with similar deletions, have been associated with higher risks of developing a more severe cardiomyopathy, suggesting different requirements between skeletal and cardiac muscles. • Dilution. As AAV does not integrate, and as muscle grows, and the number of myofiber nuclei increases between birth and early puberty, the optimal timing for using this approach requires careful assessment. • Immune responses. While AAV do not cause disease in the human, the doses that are administered systemically to these children are enormous and different types of innate and adaptive immune responses have been observed, together with the occurrence of autoimmunity against specific epitopes in rare genotypes In my presentation I will provide information regarding these key issues.

Key Words: AAV gene therapy; Duchenne muscular dystrophy; tolerability and efficacy. 1 These clinical gene therapies aim to overexpress a truncated version of dystrophin in striated muscle capable of achieving partial correction of the disease. We have examined this strategy with two different micro-dystrophins in a severe mouse model of DMD, the D2.mdx mouse, 2,3 using doses of AAV comparable to those used in the clinical trials. While significant correction of the skeletal muscle disease is observed, which slows skeletal muscle disease progression, a lethal acceleration of cardiac disease progression occurs with one of the micro-dystrophins, while the second has little or no impact on the heart. The detrimental impact on the heart appears to be caused by high levels of micro-dystrophin that result in competition between micro-dystrophin and utrophin at the cardiomyocyte membrane and by excessive protein accumulation. The micro-dystrophin with no cardiac impact does not outcompete utrophin to the same extent as the deleterious construct. While the significance of these observations for patients currently being treated with AAV-micro-dystrophin therapies is unclear, it suggests that micro-dystrophin treatments for the heart may need to be carefully titrated, and potentially dissociated from treatment for skeletal muscle, in order to achieve optimal benefits for both types of muscles. Duchenne muscular dystrophy (DMD) results from mutations in the DMD gene and the subsequent loss of functional dystrophin protein. 1, 2 In its absence, utrophin -the autosomal paralogue of dystrophin -is upregulated and compensates, albeit incompletely, for the loss of dystrophin. 3, 4 Adeno-associated virus (AAV)-mediated gene delivery provides the means to restore the expression of missing proteins with the aim of reversing the pathology of such monogenic disorders. The size of the dystrophin gene, the largest in the human genome, however, makes it impossible to re-introduce full-length dystrophin. This limitation is partly overcome by the use of heavily truncated iterations of the dystrophin protein (micro-dystrophin, μDys). 5, 6 AAVs containing versions of micro-dystrophin are now being delivered to patients with DMD in clinical trials. Such micro-dystrophins, however, are unlikely to fully compensate for the loss of dystrophin, a protein now recognized for its scaffolding/signaling roles in addition to its traditional role in mechanical force transmission. However, there has been no consideration of the fact that the cardiac disease progression is greatly slowed by the high degree of utrophin expression in the heart, 7, 8 and that there could be a displacement of utrophin by micro-dystrophin expressed at high levels. Indeed, in examining the potential therapeutic efficacy of a micro-dystrophin, μDys ΔR3-R21_ΔCT (missing spectrin-like repeats 3 through 21 as well as the C-terminal end of the dystrophin protein) 5 in the D2.mdx, 9,10 mouse model of DMD, we find that AAV-mediated overexpression of micro-dystrophin leads to the displacement of utrophin from the cardiomyocyte sarcolemma. This appears to, in essence, phenocopy dystrophin/utrophin double mutants that exhibit an exaggerated cardiac dysfunction. 7 . As the heavily truncated micro-dystrophin, μDys ΔR3-R21_ΔCT, is unlikely to functionally replace full length utrophin, its overexpression may be detrimental to cardiac function. The muscular dystrophies (MDs) are a class of genetic muscle diseases often characterized by progressive degeneration and replacement of muscle with fibrotic tissue. Limited treatment options exist for MD patients, and no therapeutics specifically target the progression of intramuscular fibrosis. NAD(P)H oxidase (NOX) 4 has been previously implicated in the development of fibrosis in other organs, but not skeletal muscle. The specific aim of the current report was to investigate NOX4 as a potential anti-fibrotic target in dystrophic skeletal muscle, using the severe D2.mdx mouse model of Duchenne muscular dystrophy (DMD). It is shown that NOX4 is increased in the muscles of D2.mdx mice and localizes primarily to interstitial regions between muscle fibers. In agreement with the hypothesis that this increase in NOX4 contributes to muscle fibrosis, genetic ablation of Nox4 prevents the progressive accumulation of fibrosis in skeletal muscle. Furthermore, it is demonstrated that pharmacological targeting of NOX4 using the small molecule clinical candidate NOX1/4 inhibitor, GKT137831, also prevents the progression of muscular fibrosis in the diaphragm and gastrocnemius of D2.mdx mice. Mechanistically, NOX4 loss-of-function results in decreased accumulation of periostin and periostin-associated interstitial cells within dystrophic muscle. These data indicate NOX4 is an effective antifibrotic target to combat progression of muscle fibrosis in DMD and, potentially, other MDs. Duchenne muscular dystrophy (DMD) is a devastating, rapidly progressive neuromuscular disease caused by mutation of the dystrophin gene and is characterized by severe muscle weakness and fatigability, early loss of ambulation and premature death. The search for effective therapies is ongoing; understanding the potential of exercise alone, or as an adjuvant to developing therapies, is likely to be beneficial but is limited by (i) a lack of understanding of exercise prescription parameters (type, intensity, target muscle groups) that are safe and effective for patients with DMD; and (ii) a lack of accessibility to exercise equipment suited to the limitations of dystrophic muscle. High intensity, eccentric actions are known to be damaging, however our group recently reported moderate-intensity isometric exercise to be safe and increase muscle strength1 in boys with DMD. Dynamic cycling was shown to benefit boys and delay loss of muscle function2,3, however improvements in muscle strength and endurance were not detected likely relating to the low intensity of the cycling exercise which as motor-assisted. Our objective was to engineer a therapeutic exercise device for patients with DMD ( Fig.  1) , allowing for safe and appropriate overload of muscle (using both isometric strengthening and aerobic cycling with assistive and resistive capabilities) to induce Spinal muscular atrophy (SMA) is a genetic disorder characterized by motor neuron degeneration and progressive muscle weakness, atrophy and premature the survival of motor neuron 1 (SMN1) gene with an incidence of 1 in 10,000 live births representing the most common genetic cause of infant mortality. Loss of motor neuron function is thought to be the initial event which triggers muscle atrophy due to the lack of physical nerve-muscle contact at the neuromuscular junction (NMJ). But, recent findings reveal that muscle atrophy in SMA precedes motor neuron death. However, the crosstalk between the muscle and the motor neuron in the pathogenesis of the disease has not yet been deciphered. We aim to clarify the motor neuron function and its contribution to skeletal muscle pathology using the multi-omics analysis of DNA epigenetics and transcriptomic profiles. Our data intriguingly supports an alternative paradigm where paracrine crosstalk between motor neuron and skeletal muscle plays a key role in the survival and function of Oculopharyngeal muscular dystrophy (OPMD) is a rare, late-onset genetic myopathy (Trollet et al., 1993) 1 affecting mainly the eyelid and pharyngeal muscles, resulting in ptosis and dysphagia, respectively. OPMD is due to an alanine expansion in the Poly-A Binding Protein Nuclear 1 (PABPN1) gene which encodes a regulator of poly-A tail length of RNA. This mutation results in PABPN1 nuclear aggregates in OPMD myonuclei that sequester essential RNA and proteins and thus induce muscle alterations. Therefore, human OPMD muscles present a global proteostatis impairment [2] [3] [4] Endoplasmic reticulum is also a key actor of cell proteostasis assuring folding of one third of the cellular proteome. Interestingly, OPMD mouse model exhibits muscle endoplasmic reticulum (ER) stress (accumulation of misfolded proteins in the ER) and restoration of the homeostatic unfolded protein response (UPR) by guanabenz acetate treatment decreases PABPN1 nuclear aggregates and improves muscle function in OPMD mice (Malerba et al., 2019) . 5 To extend these results to humans, we analyzed muscle biopsies from OPMD and healthy donors. We found that UPR genes were all upregulated in human OPMD muscles, as revealed by RNAseq analysis, strongly indicating ER stress. Consistently, GRP78 protein, the key ER chaperone, was reduced in OPMD muscles. GRP78 protein was also trapped in PABPN1 nuclear aggregates, further reducing its amount available for ER protein folding. Consequently, OPMD muscles showed an accrual of large -sheet protein aggregates (called aggresomes) in the fibers, beside to the classical PABPN1 nuclear aggregates. Consistently, in culture, OPMD human muscle cells were more likely to develop aggresomes than healthy cells following ER stress specifically. Finally, in mouse cells overexpressing PABPN1, we found that ER stress induction leads to accumulation of aggresomes and aggravation of PABPN1 nuclear aggregates. Overall, our results demonstrated an ER protein folding disruption leading to ER stress and aggresome accumulation in human OPMD muscle that could reinforce pathogenesis of OPMD. Thus, restoration of ER folding and activation of protein aggregate clearance sounds promising therapeutic strategies against OPMD. to understand gene expression regulation can provide new insights for more efficient treatments. We analyzed gene expression in gastrocnemius muscle of SOD1(G93A) ALS mouse model during pathology progression. We identified 669 mRNAs and 85 miRNAs differentially expressed. Gene Ontology analysis revealed that different cellular functions are altered in ALS such as: apoptotic processes, negative transcription regulation and nervous system development. We focused our attention on this last function identifying 3 miRNAs (miR-193a-3p, 193b-3p and 152-3p) that regulate NMJ function targeting Chrna1, Chrnb1, Chrnd, Chrne, Chrng, Emb, Musk and Prima1 (Fig. 1 ). After the in vivo modulation of these miRNAs we evidenced a reduced expression of their targets and a reduction of gastrocnemius and tibialis anterior muscle mass. In association with these observations we evidenced a fast to slow fiber shift with a reduction in the total number of NMJs ( Fig. 2 ) and in the amplitude of evocated potentials. These results support the role of 3 miRNAs in X-linked Duchenne muscular dystrophy is due to mutations in the DMD gene, which causes the almost complete loss of the membrane cytoskeletal protein dystrophin. The diaphragm is severely affected in dystrophinopathy and is characterized by progressive muscle fibre degeneration, chronic inflammation and reactive myofibrosis. The major goal of this new study was to identify novel proteomic biomarker candidates of muscle weakness and respiratory deficiency in dystrophinopathy. In order to investigate proteome-wide changes in the dystrophic and aged diaphragm muscle, our laboratories have analysed 15 months old wild type mice versus age-matched mdx-4cv mice, which represent an established genetic model of Duchenne muscular dystrophy. Mass spectrometry-based proteomics was combined with systems biological analyses and verification studies using comparative immunoblotting. A variety of changes in proteins involved in the maintenance and organization of both the extracellular matrix and the intracellular cytoskeleton were identified by mass spectrometry. These new disease-related protein markers could be helpful to improve diagnostic procedures, as well as prognostic and therapy-monitoring approaches.

Key Words: Diaphragm, Duchenne muscular dystrophy, fibrosis, mass spectrometry, proteomics. A common form of limb girdle muscular dystrophy (LGMD) is dysferlinopathy presenting as LGMDR2/2B or distal Miyoshi myopathy, that show different clinical course. They are caused by mutations in the DYSF gene encoding the protein dysferlin, a transmembrane sarcolemmal protein which has a role in the repair of sarcolemma. Dysferlin is also a t-tubule protein. To the best of our knowledge (differential) dysferlin expression in skeletal muscle fibre types has not been studied so far. We investigated by western blotting human skeletal muscles, one proximal in the arm (biceps brachii) and the other distal in the leg (tibialis anterior) which are affected differently during the course of LGMDR2: usually first tibialis anterior (around the age of 20 years) and later on the biceps brachii, with clear involvement at the age of 45 years. Muscle specimen were collected after a few hours from sudden death or muscle biopsies of biceps brachii and tibialis anterior muscle of neuromuscular controls were analysed. Dysferlin to myosin heavy chain ratio differed in biceps brachii and tibialis anterior muscle. Preliminary results show a trend towards a higher "content" of dysferlin in fast skeletal muscle, i.e. biceps brachii, compared to slow, i.e. tibialis anterior (Figure1, Table1) which might be relevant for the different course observed in LGMDR2/2B and Miyoshi myopathy. Duchenne Muscular Dystrophy (DMD) is a severe rare pediatric disease that affects the skeletal and cardiac muscles, leading to progressive muscle wasting and premature death. DMD is caused by X-linked mutations in the dystrophin gene that results in lack of dystrophin, a crucial protein involved in the biomechanical support of the muscle fibers. As a consequence of the membrane instability, DMD muscle fibers are progressively more fragile, highly susceptible to damage and fibrotic. Up to date there is no cure for DMD. Although DMD is a suitable disease for gene replacement therapy, the extent of Dystrophin transcript exceeds by far the AAVs cargo capacity and only short versions of dystrophin can be accommodated and are currently being investigating in clinical trials. However, is still not clear whether some domains of dystrophin are really dispensable for its function and stability. Our strategy consists in the generation of a larger version of dystrophin that includes additional domains required for interactions with signaling and structural proteins. To overcome the AAV limit capacity, we developed a dual system dystrophin vector, encoding for a larger dystrophin protein (Quasidystrophin) after homologous recombination of the overlapping region of the 2 dystrophin vectors. We analyzed the therapeutic efficiency of dual-AAVs gene transfer by systemic injection in DBA2-mdx mice, an aggressive DMD mouse model. Our data show high transduction efficiency in the cardiac and skeletal muscles with significant reduction in fibrosis and calcification and overall amelioration of the dystrophic phenotype using the dual-AAV vector gene transfer.

Key words: Duchenne muscular dystrophy, Dual-AAV, Dystrophin. Duchenne muscular dystrophy (DMD) is the most frequently inherited neuromuscular disease of childhood and cardiopulmonary failure is the leading cause of death for this patient population. In this study, we used electrocardiograms (EKGs) and ultra high frequency echocardiography to longitudinally monitor the heart function of a severe mouse model of DMD, the D2.mdx mouse. By 10 months of age, D2.mdx mice have a left ventricular end diastolic volume (LV EDV) that is ~32% less than age-matched wild-type mice. This restricted LV results in a significant decrease in stroke volume (SV) and cardiac output (CO); however, parameters of systolic function, such as ejection fraction (EF) and fractional shortening (FS), are preserved. In line with this impaired relaxation, EKGs of D2.mdx mice reveal they have a prolonged QT interval, due to an increase in T wave duration and indicative of impaired repolarization. Duchenne muscular dystrophy (DMD) is a severe and relentlessly progres-sive myopathy caused by out-offrame or nonsense mutations in the X-linked DMD gene. DMD is a complex disease and multiple approaches are needed to target pathological processes, both the underlying genetic mutations and the secondary complications. Despite several therapeutic options have been developed with good results, more effective treatment options are essential and may be generated through the definition of novel therapeutic targets. A muscular metabolic dysregulation is an established DMD feature with mito-chondrial dysfunctions as one of the earliest deficits that arise from multiple cellular stressors. Among metabolic regulators, Sirtuin 1 (SIRT1) represents an intriguing candidate since it acts on different aspects of cellular metabo-lism, regulating energy homeostasis, mitochondrial biogenesis, and inflam-mation. SIRT1 overexpression represents an important counter-mechanism to alleviate the dystrophic phenotype and its pharmacological modulation could be relevant as well in DMD conditions. Consistently, SIRT1 activation by selective compound, i.e., SRT2104, has already been proven to reinforce muscular structure, mitochondrial functionality, and to reduce inflammation. SRT2104 has never been tested in muscular diseases; therefore, considering its metabolic and immunomodulatory effects, we tested SRT2104 as an attractive candidate for DMD treatment. Accordingly, in our preliminary data, long-term SRT2104 administration improved muscle force and stimulated oxidative capacity. This was paralleled by reduced fibrosis and inflammatory infiltrate and increased regeneration in mdx muscle. In conclusion our results demonstrate the efficacy of SRT2104 as a new SIRT1 activator in DMD and highlight that considering DMD also as a metabolic disease and treating it as such, could provide important therapeutic strategies additional to gene therapies.

Limb Girdle Muscular Dystrophy D2 (LGMD D2) is caused by a heterozygous mutation in the termination codon of the TNPO3 gene. This mutation gives a protein which is 15-aminoacids longer in its C-terminal domain. TNPO3 gene encodes for TNPO3, which normally mediates the translocation to the nucleus of SR proteins, a family of splicing factors and other proteins related to RNA metabolism. The goal of this work was to investigate the pathogenetic mechanism of LGMD D2 through a dual approach, in vitro and in vivo, focusing mainly on the role of TNPO3 in the myogenic process and in possible muscle-specific molecular pathways. In vitro murine C2C12 myoblasts were treated with CRISPR/Cas9 genome editing of TNPO3. Preliminary data suggest morphological, proliferative and expression changes of genes involved in myogenic differentiation in comparison to the C2C12 control line (Fig.1 ). For the in vivo approach, the microinjection of mRNA encoding human wild type (WT) and mutated TNPO3 in zebrafish embryos allowed to study its effects during embryonic development. We monitored the gene expression profiles of endogenous transportin 3, of myogenic regulatory factors (MRFs) and muscle-specific proteins, suggesting morphological and gene expression changes in embryos miscroinjected with the WT form and particularly in microinjected embryos with the mutated TNPO3 form. The approaches used are a first step to understand the role of TNPO3 in muscle physiology and in the pathogenetic mechanism underlying LGMD D2 which is still unknown. Adipose tissue can accumulate between muscle fibers, known as intramuscular fat (IMAT) and is strongly correlated with loss in muscle mass and function. IMAT infiltration is a hallmark of ageing and a diverse set of diseases such as muscular dystrophies, diabetes, and spinal cord injuries. No therapies exist currently that limit IMAT formation. We and others have shown that exogenous activation of the Hedgehog (Hh) pathway is beneficial for overall muscle regeneration and improves muscle health in a Duchenne muscular dystrophy (DMD) model (mdx mouse). We have also demonstrated that pharmacological and genetic activation of the Hh pathway blocks the formation of IMAT after an adipogenic injury and in mdx mice. To define the endogenous function of the Hh pathway during acute muscle regeneration, we evaluated which of its 3 possible ligands ( Skeletal muscle circadian clocks regulate a daily program of gene expression that is necessary for predictive homeostasis and tissue resilience. Aging is associated with progressive decline in skeletal muscle function. To date, our lab has found age associated changes in expression of the core clock gene, PER2, in mouse muscle. However very little is known about how aging alters the circadian transcriptome. To address this gap in the field, we collected skeletal muscle from Young (6mo), Aged (18mo), and Old (27mo) male C57BL/6J mice every 4h for 48h for RNA sequencing and circadian analysis. We identified that circadian expression of the core clock genes was unchanged from Young to Aged, however, from Aged to Old there were significant changes in several genes within the repressor limb (e.g. Per1/2, and Cry1/2). Consistent with age-associated changes in core clock genes we found an age-related decline in the circadian transcriptome that was evident by 18 months, with 941 oscillating genes in the Young mice, 699 oscillating genes in the Aged mice, and 474 oscillating genes in the Old mice. Comparing across the 3 ages there was less than 10% overlap in the circadian transcriptome between any age and only 3% of genes shared across all three ages, and these shared genes were composed primarily of the core circadian clock factors.

These are the first data to demonstrate a progressive agerelated decline the skeletal muscle clock and the skeletal muscle circadian transcriptome. Functional cluster analysis of the transcriptomes suggest that altered Mitochondrial and metabolic disorders such as heart failure with preserved ejection fraction (HFpEF) cause skeletal muscle contractile dysfunction through mechanisms not fully understood. Abnormal mitochondrial metabolism causes cytosolic accumulation of metabolites of the tricarboxylic acid (TCA) cycle, such as succinate and its derivatives. 1 Therefore, we investigated whether succinate impairs skeletal muscle contractile function. We exposed mouse diaphragm bundles to dimethyl-succinate (0.5 to 10 mM, membranepermeable) ex vivo for 1h to 4h at 37°C and measured contractile properties. Dimethyl-succinate (0.5 and 1 mM, 4 h) decreased diaphragm twitch force (~15%), tetanic force (~10%), and peak power (~25%; Figure  1 .A). The greater decline in peak twitch force compared to maximum tetanic force allude to impairments in the excitation-contraction coupling (ECC) machinery, whereas the notable decrease in peak power implicates dysfunction in sarcomeric proteins. Myofibrillar and calcium-handling proteins are susceptible to hypersuccinylation at lysine residues, suggesting that elevated lysine succinylation contributes to the contractile impairments in conditions associated with succinate accumulation. We performed post-translation modification proteomics in diaphragm bundles exposed to dimethyl-succinate (10 mM, 1h: 75-90% decrease in force and power) and found increased lysine succinylation in nebulin (6-fold), titin (5-fold) and troponin-I (2-fold). Myosin heavy chain (fast), myosinbinding protein C (fast), troponin C, myomesin, and calsequestrin were among the proteins with lysine succinylation detected only in bundles exposed to dimethyl-succinate. Inhibition of complex II activity (dimethyl-malonate, 10 mM) had no effect on contractile function and did not prevent the loss of force and power induced by dimethyl-succinate. Exposure of permeabilized single fibers to dimethyl-succinate (10mM, 1h at 15°C) revealed a ~20% decrease in maximum isometric force in fast-twitch fibers and a ~20% decrease in submaximal force and calcium sensitivity in slow-twitch fibers. The high concentration used in single fibers accelerates mass action and elicits non-enzymatic effects 2 suggesting direct effects of succinate on sarcomeric proteins. Our study shows that membrane-permeable succinate causes isometric and isotonic contractile dysfunction via impairments in excitation-contraction coupling and sarcomeric protein function. Moreover, several myofibrillar proteins responsible for sarcomere structure and function emerged as being susceptible to lysine succinylation. The data suggest that cytosolic accumulation of succinate is a potential mediator of contractile dysfunction in mitochondria and metabolic disorders. Insulin-like growth factor I (IGF-I) contributes to glucose homeostasis and may glucose handling in skeletal muscle through activation of hybrid receptors, which contain insulin and IGF-I hemireceptors. Our previous work showed that loss of IGF-I in murine muscle can have negative effects on glucose metabolism and exercise tolerance in addition to muscle mass. AMP-activated protein kinase (AMPK) has a key role in maintaining the balance between anabolic and catabolic programs for cellular homeostasis in response to metabolic stress.Increased AMP/ATP ratio in muscle cells during repetitively contraction activates phosphorylation at AMPKα Thr172 , and this induces downstream signaling cascades to promote glucose uptake. Phosphorylation of AMPKα Ser485/491 can inhibit AMPK activity and insulin may be one of its upstream triggers in skeletal muscle. The influence of IGF-I in glucose uptake and its interplay with AMPK activation during exercise is not well understood. The aim of this study is to determine the acute response of AMPK signaling pathway after a single bout of aerobic exercise in muscle loss or overexpression of IGF-I ( Figure 1 ). We used two different mouse models in our study to determine the acute exercise response in muscle deficient or overexpressed IGF-I: Muscle IGF-I Deficient (MID) and mice harboring the rat Igf1a cDNA under the control of the fast myosin light chain promoter (mIgf1 +/+ , MLC/mIgf). We induced Igf1 deletion with chow containing doxycycline at postnatal day 21 for 5 days in MID and the controls without floxed exon 4 of Igf1 (Igf1 fl/fl ). Littermates without the transgene were used as wildtype controls for mIgf1 +/+ mice. We used 16-weekold MID and mIgf1 +/+ transgenic male mice, and relevant controls in our study. The mice were randomly assigned to No Exercise control (NO) or to exercise groups with three time points (0-hour, 2-hour, or 24-hour after exercise). For the exercise groups, mice ran at 20 m/min with 5% inclination for 30 minutes. Tibialis anterior muscles from each mouse were used to examine AMPKα phosphorylation. In our preliminary data, the phosphorylation of AMPKα Ser485/491 was higher at rest in mIgf1 +/+ transgenic muscle compared to wild type muscle, and the trend of phosphorylation was different immediately after exercise and 2-hours later. In contrast, phospho-AMPKα Ser485/491 seemed unchanged in MID mice. This suggests that high IGF-I levels may be needed -52 -sustaining heme biosynthesis. Being part of the heme synthesis-export functional axis, heme export by FLVCR1a has been reported to sustain the tricarboxylic acid cycle (TCA) flux and the electron transport chain (ETC) activity in tumors 3 . Here we generate skeletal muscle-specific Flvcr1a-null mice and analyze the impact of disrupted heme synthesis-export axis in this tissue. Metabolic data obtained in gastrocnemius homogenate show that, upon deletion of Flvcr1a, the activity of TCA cycle enzymes and ETC complexes is increased, along with glutaminolysis and fatty-acid beta oxidation. Conversely, the activity of glycolytic enzymes is reduced. Flvcr1a deletion also affects muscle morphology. The distribution of cross-sectional fibers' area (CSAs) is compromised in Flvcr1a knockout mice, showing a prevalence of fibers with small area. Along with these data, increased number of PAX7+ cells was also observed. In motor behavior tests, Flvcr1a knockout mice show worse performance compared to controls, a phenotype exacerbated by aging. Collectively, these results hint that FLVCR1a is involved in the regulation of energetic metabolism in skeletal muscle, and that the heme synthesis export system is important to maintain proper muscle function. Despite data over several decades showing that resistance exercise induces hypertrophy, the transcriptional mechanisms responsible for the process are still not fully understood. Recent meta-analyses have attempted to identify time-dependent patterns in the expression of genes that regulate the transcriptional response to resistance exercise. 1, 2 The objective of this study was to perform a detailed transcriptomic analysis through RNAsequencing of the acute response to resistance exercise (one hour after an exercise session), across a 30 day -53 -period of daily resistance training. We have developed and implemented a highly controllable resistance training model that uses implantable pulse generators in freeliving rats to induce hypertrophy in the tibialis anterior by co-contraction of the dorsiflexors and plantarflexors (3) . We assessed the skeletal muscle transcriptome (1hour post exercise) after 2, 10, 20 and 30 days of our exercise training paradigm and found 5 temporal clusters across our timecourse (Self-organizing maps (SOM) 1-5, Figure 1B ). SOM1 and SOM2 included known resistance-exercise responsive pathways including the ribosome, proteasome, and protein processing in the endoplasmic reticulum as well as genes associated with inflammation/macrophages. These genes were highly responsive while the muscles were growing, but their expression was blunted as muscle mass reached a new steady state. GeneMANIA network association analysis (Figure 1c) found that the genes within these clusters were centrally controlled by the transcription factor Myc, a well-known regulator of cell cycle, metabolism, growth, RNA polymerase activity and ribosomal biogenesis (4). However, the expression of many of these genes in skeletal muscle is not well understood. We propose that it will be informative to study the group of genes whose timecourse of expression is associated with the period of growth and whose response declines as the muscle adapts to a daily stimulus to achieve a new steady state. The skeletal muscle tissue is a primary metabolic activity site controlling almost 80% of insulindependent glucose uptake and storage. It is also the largest protein reservoir (75%) of our body, supplying aminoacids to support energy production in other organs during catabolic conditions. Thus, skeletal muscle is crucially important to control health and disease progression in distant tissues both for its role in balancing other organs' metabolic needs and its reserves -54 -to use in energy production. Its dysfunction is associated with poor prognosis and reduced quality of life. Peroxisomes are ubiquitous dynamic metabolic organelles that play an essential role in a variety of cellular events such as reactive oxygen species clearance and lipid metabolism. However, their role in muscle metabolism and function has largely been overlooked. To investigate the consequences of peroxisomal dysfunction in muscle we have generated a mouse model with impaired peroxisome biogenesis by disrupting specifically in skeletal muscle the Peroxisomal biogenesis factor 5 (PEX5), which serves as a shuttle receptor for the import of peroxisome matrix protein. We found the peroxisomal dysfunction in muscle causes several mitochondrial defects, and increased ROS production resulting in muscle ultrastructural alterations and reduced running performance. Thus, peroxisomal integrity is critical for skeletal muscle homeostasis. Key Words: Peroxisomes; skeletal muscle; ROS.

A healthy mitochondrial network is essential for postmitotic tissues as muscles. Mitochondria-shaping machinery is downregulated in sarcopenia and is maintained by lifelong exercise. OPA1 is a profusion protein that plays an important role in mitochondrial dynamics we have demonstrated. Interesting, OPA1 protein is significantly decreased in sedentary humans and old mice compared to young controls and, exercise is sufficient to restore the protein level. In conditional tissue-specific Opa1 ko is sufficient to induce a sarcopenic-cachectic phenotype in just 3 months of deletion and Fgf21 is a central myokine in this process. This work aims to find new specific players involved in aging sarcopenia. Particularly, that synergistically promotes the aging of the whole body and the secretome related. we are comparing different genetic mouse models to dissect the specific role of the different genes in the sarcopenia scenario. Mitochondrial dysfunction in muscle tissue is sufficient to directly drive metabolic changes and systemic inflammatory by increasing the expression and secretion of the myokines FGF21 and IL6. Despite the key role of FGF21, we have new evidence that is connecting mitochondrial dysfunction, autophagy alteration, muscular IL6, and aging. Opa1 is a "sensor" for the health of the muscle, the reduction of the protein level recapitulates the acute sarcopenia process. -55 -

The changes related to mitochondrial dysfunction are specifically correlated to the shape of these organelles. Different alteration induces some common but mainly specific metabolic and stress responses directly correlated to sarcopenia development. Transcription Factor EB (TFEB) is a master metabolic regulator in skeletal muscle, finely tuning fuel uptake to energy production during exercise. These findings strongly support TFEB activity as crucial for mediating the beneficial effects of exercise1. Starting from this evidence we decided to discover new uncharacterized genes that may mechanistically explain adaptive effects of TFEB on skeletal muscle. By crossing gene expression data of TFEB transgenic and trained muscles, we found a gene of unknown function belonging to the Riken cDNA collection2 that we called Exe-Riken. We showed that Exe-Riken is a real protein coding gene. We also confirmed its transcriptional dependence from TFEB and exercise, with a fine tuning in different muscles; interestingly, its transcription induction does not depend on extra muscular factors. Moreover, we showed its differential localization in muscle fibers, with a positive correlation between cross sectional area and immunostaining reactivity. Oxidative muscles such as soleus did not present cytoplasmic but more nuclear Exe-Riken localization. Cancer cachexia is a multi-factorial metabolic syndrome characterized by excessive body weight loss due to progressive fat and lean mass catabolism in spite of nutritional support, 1 occurring in more than half of cancer patients. 2 The frailty experienced by cachectic patients increases morbidity, reduces tolerance to treatments, complicates patient management and accounts for up to 30% of cancer deaths. 3, 4 The loss of skeletal muscle mass and strength are considered as the most relevant features of cancer cachexia and predictors of poor outcomes. 5, 6 Bone Morphogenetic Protein (BMP)/Smad1/5/8 pathway is a positive regulator of muscle mass repressing the transcription of MUSA1, an atrophy-associated E3 ubiquitin ligase that facilitates proteasome-dependent protein breakdown. 7, 8 We demonstrated that diminished BMP (Bone and Morphogenetic Protein)/Smad1/5/8 signalling in muscles plays a critical role for the onset of cancer cachexia. Re-activation of BMP signaling in muscles is sufficient to prevent tumor-induced muscle atrophy, and to extend the lifespan in tumor bearing mice despite tumor growth. Importantly, these protective effects are prevented by knocking down Smad1/5. 9 We aim to understand how the cancer stimulates transcriptional changes leading to BMP-Smad1/5/8 inhibition in skeletal muscles to favor cachexia onset in order to identify new potential therapeutic targets. Sarcoglycanopathies are recessive forms of muscular dystrophies in which the disruption of the sarcoglycan complex results in sarcolemma fragility and progressive muscle degeneration. Most of the reported cases are due to missense mutations originating a folding-defective sarcoglycan (SG) eliminated by the cells' quality control system, although potentially functional. 1 No treatment for sarcoglycanopathy is currently available. Cell and gene therapy-based approaches are under investigation, even if many challenges are present. Among others, there is the difficult of delivering the gene to skeletal muscle, which is the most abundant tissue of human body, the problem of immunogenicity of the viral vector and of the transgene (if the protein is totally absent in the patient), etc. An interesting alternative could be represented by a class of small molecules, called protein folding correctors, able to recover the mutants and avoid complex disruption. Furthermore, small molecules are easier to formulate, deliver and eventually optimize. Particularly, CFTR correctors are a subgroup of cystic fibrosis modulators able to enhance or even restore, through different mechanisms, the expression, function, and stability of type II CFTR mutants. 2 In α-sarcoglycanopathy, the effective rescue of different SG-mutants has been proved for a few of such molecules by using cell models and, importantly, myogenic cells from patients. 3, 4 In addition, their pharmacological efficacy has been successfully tested in novel mouse models, ad hoc generated, expressing the human mutated α-SG sequence in both hind-limbs, delivered by early AAV injection. As the proof of concept of the strategy has been established for LGMDR3 both in vitro and in vivo, we checked the broadness of such approach by testing the effect of a panel of different CFTR correctors in LGMDR4 patient's myotubes, carrying a missense mutation on the SGCB gene. The analysis revealed the increased expression of the mutated β-SG protein upon treatment with the C17 molecule, the most promising corrector in our hands. In particular, C17 induced a twofold increase in the β-SG membrane fraction in comparison to vehicle treated cells. The myotubes surface re-localization of the β-SG protein was further -59 -proven by immunofluorescence staining. These results confirm the hypothesis that CFTR correctors could be in principle applied to all the sarcoglycanopathies cases linked to missense mutations, regardless of the SGC gene. Remarkably, these data are the proof that CFTR correctors are effective not only in cystic fibrosis, but also in sarcoglycanopathy, suggesting a great potential for the protein conformational diseases, especially for those orphan diseases where no current therapeutics are available. and/or DOX prior to unilateral Cardiotoxin (CTX) injection into the TA(3), while the contralateral TA served as an internal control. Muscles were harvested at days 3, 5, 7, 11, and 14 post-CTX. TA masses from all groups at days 3-7 post-CTX were significantly lower (p<0.05) compared to contralateral non-damaged muscles. All groups were able to recover to non-damaged mass levels by day 14 post-CTX except SID animals. There were no differences when comparing CTRL and MID muscle fiber areas, instead, SID muscle fibers were smaller when compared to CTRL muscles, suggesting that satellite cell IGF-I are crucial for muscle regeneration. Surprisingly, SMID mice were able to recover to CTRL levels in fiber area, indicating a potential compensatory IGF-I supply from non-muscle sources. Next, in-situ functional testing(4) was performed comparing damaged vs. non-damaged TAs. At day 14 post-injury, normalized forces were lower in damaged TAs of CTRL, MID, and SMID animals when compared to contralateral non-damaged TA; however, there was no difference in force for SID animals, suggesting the importance of muscle-specific IGF-I for muscle function post-injury. In conclusion, IGF-I generated by satellite cells is critical for efficient skeletal muscle regeneration, while both satellite cells and myofibers provide integral sources of IGF-I for muscle development.

Key Words: Satellite cells; IGF-I; muscle regeneration; postnatal muscle growth.

Hotel Petrarca, Montegrotto, Euganean Hills, (Padua), Italy Energy balance maintenance is crucial to preserve astronaut's wellness during longduration-spaceflight. Nutritional intervention can counteract the detrimental effect of microgravity on body composition and metabolism. Methods and procedures: NutrISS is an Italian Space Agency biomedical experiment. This proofof-concept study aim to monitor the body composition of astronauts and to provide nutritional advice during longterm mission. The study has been executed on the ISS and sponsored by ESA. The payloads selected for both missions (i.e. "BEYOND" and "COSMIC KISS") result from a public-call funded and coordinated by ASI, in the Oxidative metabolism is responsible for most of ATP resynthesis during work, leisure or sport activities lasting longer than 1-2 minutes. A substantial impairment of oxidative metabolism during exercise is known to occur following inactivity. Whereas the "central" (mainly cardiovascular) factors contributing to this impairment have been thoroughly investigated, "peripheral"

impairments (related to microvascular and endothelial functions, intramuscular O2 delivery-O2 uptake coupling, peripheral O2 diffusion, skeletal muscle oxidative metabolism, mitochondrial respiration) have received relatively less attention. We recently investigated several central and peripheral factors potentially responsible for an impairment of oxidative metabolism during exercise in 10 young males undergoing 10 days of horizontal bed rest (BR) ( In conclusion, peripheral limitations to oxidative metabolism after a 10-day BR were "upstream" of mitochondrial function, at the level of microvascular O2 delivery/endothelial function, and at the intramuscular matching between O2 delivery and O2 uptake. On the other hand, mitochondrial content and mitochondrial respiration were unaffected or even improved (enhanced mitochondrial sensitivity to submaximal [ADP]). The decreased resting muscle V O2 following BR could represent an adaptive phenomenon in response to simulated microgravity-inactivity, attributable to the fact that muscle catabolic processes within muscles are less expensive, in terms of energy, than anabolic ones. These concepts, besides being of interest from a basic science point of view, may be relevant for patients with pathological conditions characterized by relatively short periods of profound inactivity, and could affect the definition of countermeasures or rehabilitative interventions. Disuse induced loss of muscle mass is a frequent phenomenon and a very relevant clinical problem. It exposes subjects to falls and fractures and favour further deconditioning. It is related to metabolic alterations, such as insulin resistance, and to low grade chronic inflammation which are among the major risk factors of chronic diseases. It worsens the prognosis of many chronic diseases 1, which benefit from exercise training. We studied structural, functional and proteomic adaptations of skeletal muscle and the underlying cellular and molecular mechanisms induced by spaceflight in humans and by simulated microgravity both in humans, bed rest (BR) and unilateral lower limb suspension (ULLS), and in mice, hindlimb unloading (HU). As expected, in both human and mice models we found muscle atrophy and a slow to fast myosin isoform shift. Moreover, a downregulation of most myofibrillar proteins was observed by 2D proteomic and of myosin by quantitative one-dimensional electrophoresis. To address the mechanisms of such adaptations, we studied intracellular signalling pathways controlling muscle mass and metabolism. In human bed rest and mice HU, we observed: upregulation of NRF2, a transcription factor sensing reactive oxygen species (ROS) in the cell; adaptations of major ROS buffering systems (e.g., SOD, catalase, peroxiredoxins, Hsp); protein oxidation. Such findings were consistent with a potential role of redox imbalance in causing muscle wasting as suggested by several previous findings. To test such hypothesis, we administered trolox, a potent antioxidant, to HU mice. Importantly, NRF2 was not up-regulated and no protein oxidation was found, indicating that redox imbalance was prevented by trolox. However, expressions of MuRF-1 and atrogin-1, major ubiquitin ligases of the ubiquitin proteasome system, were still up-regulated and muscle atrophy was not prevented by trolox. Therefore, redox imbalance was not likely to cause muscle atrophy, at least in mice. Importantly, in human BR and ULLS and in mice HU, proteomic analysis indicated downregulation of both aerobic and anaerobic metabolic enzymes, i.e., a general derangement of energy metabolism. In human BR and mice HU, PGC1α, an activator of transcription controlling mitochondrial dynamics and biogenesis and inhibiting the ubiquitin proteasome system, was downregulated and expression of proteins involved in mitochondrial dynamics was altered. On such ground, we hypothesized that a metabolic program of muscle atrophy could be activated in disuse. To test such hypothesis, we subjected transgenic mice selectively overexpressing PGC1α in muscle to HU. In both soleus and gastrocnemius muscles of such mice, no upregulation of atrogenes expression was observed following HU and muscle atrophy was almost completely prevented confirming a major role of a metabolic program in disuse muscle atrophy. Exercise is an attractive tool to prevent muscle wasting as it is known to cause mitochondrial adaptations and to enhance PCG1α expression. Few, but encouraging evidences on the use of exercise as a tool to prevent disuse atrophy were published. The mechanism of exercise-induced protection on muscle atrophy appears not clearly defined yet. We will present novel data on the mechanisms underlying prevention of muscle atrophy by exercise preconditioning.

A reduction in skeletal muscle activation (disuse), regardless the cause (i.e.: physical inactivity, disease and microgravity), can induce profound functional transformations affecting the metabolic, and neuromuscular properties of the the muscle tissue. It is also well known, both from animal and human studies, that although the main alteration caused by disuse is represented by muscle atrophy, there is evidence that similarly the quality of the muscle is altered, as demonstrated by the fact that the loss of muscle strength is considerably greater than that of muscle size. 1 The reduction of muscle quality can be ascribed to a complex interaction of various factors affecting neuromuscular transmission, 1 muscle architecture, 2 and contractile apparatus properties. 3 Short-term disuse is a common condition experienced by those individuals, that are bedridden because of an injury, surgery and/or chronic medical conditions. In the present study we focused our attention on the functional skeletal muscle alterations induced by short term disuse, consisting of 10 days of unilateral lower limb suspension (ULLS, walking with crutches), and the effect of a retraining period on the contractile properties of neuromuscular system investigated at the level of the single motor units (MUs). To this purpose, the electrical activity of the muscle vastus lateralis of 11 young males (22.1 ± 2.9 years) was recorded with high-density electromyography (HD-EMG), using a grid of 64 electrodes (5 columns × 13 rows; gold-coated; diameter of 1 mm; inter-electrode distance of 8 mm), during isometric trapezoidal contractions executed at 10, 25 and 50% of maximum voluntary contraction (MVC) and characterized by 3 phases: a linear increase (recruitment phase) in force at 5% MVC/s, a 20 s constant force at target (plateau phase) and a linear decrease (de-recruitment phase) in force at 5% MVC/s. These measures were performed at baseline (BLS0), and repeated immediately after the suspension period (LS10) and after 21 days of retraining (RT1). As expected, MVC decreased at LS10 (from 816.18 ± 97.22 to 573.20 ± 104.69 N; p<0.001) as did the ability of the participants to maintain the constancy (steadiness) of the isometric contraction, during the plateau phase, with a reduction observed at 25% MVC. After the HD-EMG decomposition and the removal of duplicated MUs, a total of 1422 unique MUs were identified. Moreover, we observed a reduction in absolute MUs recruitment and de-recruitment thresholds at all intensity of contractions. The MUs discharge rate decreased at 10 and 25% MVC, at all the points of evaluation (e.g., recruitment, de- recruitment and plateau), but not at 50%. The coefficient of variation of MUs interspike interval (COVisi), a marker of irregular firing pattern, increased during the steady state phase at 25% MVC. After the 21 days of retraining, MVC, recruitment threshold and discharge rate recovered to baseline levels while the COVisi and steadiness improved compared to baseline. In conclusion, these results highlight that 10 days of ULLS can induce significant alterations of the skeletal muscle function at the level of the single MUs and that 21 days of resistance training can restore these impairments. Additionally, there is a paucity of knowledge about molecular pathways dysregulated in aged motor neurons and PSCs. In this talk, I will present compelling evidence that aging does not result in the loss of motor neurons. I will instead show that motor neurons undergo subcellular and molecular changes during aging. These include loss of synaptic inputs and altered expression of genes with potential roles in synaptic maintenance and repair. I will also present data showing that PSCs accrue a myriad of age-related cellular and molecular features but do not decrease in number with advancing age. I will end by describing studies needed to further advance our understanding of aging NMJs, and its contribution to sarcopenia. The progressive loss of muscle mass (sarcopenia) leads to serious consequences such as a loss of mobility and autonomy. 1 Deterioration of the neuromuscular junction (NMJ) is an emerging factor in the development of sarcopenia. The JNM is a tripartite structure, composed of the presynaptic terminal, the postsynaptic element and by glial cells which are the perisynaptic Schwann cells (PSCs). PSCs regulate the maintenance, remodeling, and repair of JNMs in mouse models, 2 functions that are altered in aging purportedly to facilitate the reinnervation of the denervated NMJs. 3, 4 However, not only is human NMJ poorly characterized, but PSCs have never been investigated despite the essential role attributed to them in mice. Here we report the characteristic of NMJs at young and older active males. NMJ-enriched biopsies were performed using the BeeNMJ method5. In young individuals, the NMJ innervation was robust and showed little variability as no denervation was observed and only a small percentage of partially innervated NMJs was observed. In contrast, in older individuals, we observed more variability between individuals, showing a significantly larger percentage of partial innervation. We will discuss the relationship between the innervation status and the physiological performances of the individuals Key Words: Human neuromuscular junction; perisynaptic Schwann cells; aging.

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Department of Physical Therapy & Myology Institute, University of Florida, Gainesville, FL, USA. Russell T. Hepple, Email: rthepple@phhp.ufl.edu

Denervation of skeletal muscle fibers is amongst the most influential processes in aging skeletal muscles. It contributes to various aging muscle phenotypes, including muscle atrophy and fiber type shift. Notably, skeletal muscle denervation with aging can be exacerbated in various conditions, such as chronic obstructive pulmonary disease (COPD) 1 and physical frailty, 2 leading to an acceleration of muscle atrophy, weakness, and mobility impairment. COPD is most often precipitated by years of heavy smoking and interestingly, previous studies have shown that smoking can yield marked fiber type shift even in the absence of lung disease, consistent with the established impact of denervation. Furthermore, smoking accelerates muscle atrophy with aging, 3 and has recently been shown to alter neuromuscular junction morphology. 1 Physical frailty is another condition associated with exacerbated denervation and more severe muscle atrophy with aging, and recent studies link this with an elevated blood level of metabolites of tryptophan, such as kynurenine. 4 Interestingly, motoneurons in culture exposed to kynurenine exhibit axonal degeneration, 4 which could be a contributing mechanism to the exacerbated atrophy and muscle dysfunction in the frail elderly. In seeking a mechanistic explanation for the neuromuscular junction alterations seen with chronic tobacco smoke and kynurenine, we note that both cigarette smoke and kynurenine stimulate the aryl hydrocarbon receptor (AHR). The AHR is a ligand-activated transcription factor that upregulates detoxifying pathways which include cytochrome P450 enzymes (e.g., Cyp1A1) in response to a wide variety of endogenous and exogenous chemicals, and whereas short-term AHR activation can be adaptive, chronic activation can lead to toxicity via mitochondrial mechanisms. Notably, we find that knockin of a constitutively active mutant of the AHR in skeletal muscle also causes neuromuscular junction alterations, 5 suggesting chronic AHR activation may contribute to the exacerbated denervation seen in COPD and physical frailty. Future studies that manipulate AHR activation (e.g., via AHR inhibition or knockout) will be valuable in identifying whether AHR-targeting therapies may be beneficial in attenuating exacerbated denervation in aging muscle. Key Words: Aging; coexpression of fast and slow myosin heavy chains, denervation and reinnervation, fiber-type grouping, Human skeletal muscle; recreational sport activity. Several human studies have demonstrated an age-related reduction in the number of motor units (MU), yet the ability of a MU to undergo remodelling and to expand to rescue denervated muscle fibres is also a defining feature of their plasticity. Remodelling of MUs involves two coordinated processes; i) axonal sprouting and new branching growth from adjacent surviving neurons, and ii) the formation of key structures around the neuromuscular junction (NMJ) to resume muscle-nerve communication. Exercise in older age has demonstrated benefits in multiple physiological systems including skeletal muscle and Masters Athletes may be viewed as an effective model to study the effects of ageing dissociated from the effects of reduced activity. Several lines of evidence also suggest exercise into older age may also extend to peripheral MU remodelling. There is, however, a lack of research in humans due to methodological limitations which are easily surmountable in animal models. As such, the ability to accurately quantify remodelling and relate it to clinically relevant neuromuscular function has proven difficult. This talk will address the neuromuscular function and MU structure and function of elite Masters Athletes competing in endurance and power events, in comparison to their less-active counterparts, and crucially, to frail older adults. Although advanced age is typically associated with poor physical function that can adversely affect mobility and quality of life, some individuals retain high function well into their advanced years and can provide clues to mechanisms of healthy muscle aging. With this in mind, we have been conducting extensive studies of competitive octogenarian athletes. Herein, we present data on 15 world class track and field masters athletes (MA; 7 female), 8 of whom were the current world record holders for their age and discipline at the time of study, in comparison to a group of 14 age-and sex-matched healthy non-athlete individuals (NA; 6 female). As expected, the MA group had superior physical function on a variety of clinical function tests (e.g., 4 m fast walking speed, cardiopulmonary exercise testing to VO2peak, isokinetic knee extensor strength), and this was associated with higher thigh cross-sectional area, higher motor unit number estimates, smaller accumulation of very small muscle fibers, less denervation, fewer oxidatively compromised muscle fibers, and higher mtDNA copy number. Muscle proteomics analysis revealed that MA muscle was characterized by over-representation of proteins involved in mitochondrial biology (nuclear and mitochondrial DNA encoded proteins, mitochondrial Sirtuins, mitochondrial dynamics, mitochondrial cristae remodeling, mitochondrial proteostasis), and an underrepresentation of proteins involved with nuclear pore complexes and the spliceosome. Interestingly, when comparing our MA to previous studies examining exercise-responsive proteins, 1 and age-responsive proteins, 2 although there was substantial overlap between proteins over-represented in MA and those elevated in subjects with higher physical activity, there were 301 proteins that were uniquely over-represented in MA that had not previously been identified as responsive to exercise. As such, our results suggest that a high level of physical activity is likely not the only factor explaining the superior function in MA and provides exciting insights to novel mechanisms for promoting healthy muscle aging. Key Words: Aging; skeletal muscle; mitochondria, exercise, masters athlete. The number of marathon events increased during the last 30 years, and in particular increased those of the Master Athletes. 1, 2 There are evidences that long-distance running could provide considerable health benefits for older runners, specifically risk reduction of cardiovascular diseases, cancer, diabetes, depression, and falls. 3, 4 In this study we have analyzed data from several editions of the Venice marathon, a famous Italian race that attracts people from every corner of the world. The data analysis was done on age category distribution (left panel), nation (right panel) and performance in 13 editions (2007-2019) evaluating the performance of all the athletes vs. the top 10% fastest athletes (TOP 10%). The analysis has shown a steady increase in female participation, in particular for 40-50 years old, and a substantial reduction of performance in the younger category of 30-35 years old, both male and female, from 2003 to 2019. The intra-category differences of performances have shown that the trends in male and female were similar and a specific steeper decline in performance was identified at age 55 for the women and -70 -at 50 for the men with specific differences between the set of all athletes and the TOP10% one. The model of the Venice marathon can therefore be useful to analyze the gender participation and in particular the human model of successful aging . [5] [6] [7] Key Words: Aging performance decay; Masters; gender differences.

Paolo Gava, Barbara Ravara, Ugo Carraro Aging behaviors differ in females and males. Females are weaker, but survive longer. Paolo Gava described the gender decay of muscle performance by comparing the world records of Masters, who compete in age groups of 5 years from 35 to 100 or older. After normalizing -that is, taking the all-time world record of young sports athletes to 100% -the women's and men's world records are lists of 16 data showing declining trends in muscle performance as aging, indicating only minimal gender differences in the process. All trend lines tend to zero at around 110 years. The decline of short running disciplines is very similar. It is smooth from 30 to 50 years, it is almost linear from 50 to 70 years, then the decay is progressively steeper. Since gender hormones appear to have little influence on the decay of aging performance, other key bioregulators, such as those of cellular energy metabolism, appear to be more relevant. Prior studies of isometric contraction in calf muscle using velocity encoded phase contrast (VE-PC) imaging have revealed several interesting features in the medial gastrocnemius including strain heterogeneity along and across fibers, gear ratios decreasing with unloading, and constant in-plane areas during isometric contraction suggesting little or no deformation in the out-plane direction 1,2 . In addition to analyzing fiber strains, spatially localized strain and strain rate (SR) tensors can be computed from velocity encoded phase contrast images that have the potential to identify changes in tissue deformation along and perpendicular to the muscle fiber. Strain tensor analysis has revealed anisotropy of deformation in the cross-section of the muscle fiber, deviation of the principal strain direction from the muscle fiber orientation, heterogeneity of strains along the length of the muscle and across the muscle fiber and in study of age-related effects, maximum shear strain emerging as the most significant predictor of force loss with age 3 . Earlier studies identified fiber direction from fascicle directions on water suppressed MR images (fascicles appear bright due to fat infiltration at the fascicles). -72 -However, identification of fascicles in young subjects is difficult since the fascicles are relatively thin and do not have sufficient fat infiltration. We used the average of the lead DTI eigenvector in each region to extract the direction of fibers. Fiber strains, length and angle changes were calculated for three-foot positions and at two % MVC at each position. This study calculates regional fiber strains, and strain and strain rate (SR) tensor in the medial gastrocnemius muscle from 2D velocity encoded phase contrast (VE-PC) images acquired under isometric contraction at three-foot positions (representing different muscle fiber pennation angles/length) at two % MVC's at each position. The plantarflexed foot position had the highest normalized fiber strain while the dorsiflexed position had the lowest normalized fiber strains (to force and to torque) ( Table  1 ). In the strain and strain rate tensor analysis, plantarflexed foot position had the highest normalized strain and strain rate (to force) and strains decreased with %MVC's compared to the neutral and dorsiflexed foot positions ( Figure 1 ). Strain in the fiber cross-section showed radial expansion in the in-plane and radial contraction in the out-plane direction; radial deformation was lowest in the plantarflexed position.

Key Words: Dynamic muscle MRI; fiber architecture and function; fiber strain; strain tensor mapping.

Prior studies of isometric contraction in calf muscle using velocity encoded phase contrast (VE-PC) imaging have used either single slice imaging to extract 2D strain and strain rate tensors 1,2 or sequentially acquired single slice images which were concatenated to form a volume to extract 3D strain and strain rate tensors 3 . The advantage of 2D VE-PC is that it is fast but requires one to acquire the images in the plane of the fibers to leverage the fact that muscles exhibit planar deformation; a 3D acquisition circumvents the need for this positioning. A volume acquisition VE-PC sequence is usually too long for subjects to maintain consistent contractions through the scan duration. In this study, we present (i) sequentially acquired anatomical slices to extract 3D strain tensor and projections on the myofibrillar aggregate in a cohort of young and old subjects along with correlation of the strain indices to histology and (ii) initial results using a highly accelerated 4D Compressed Sensing flow sequence 4 to image the much smaller motion seen in muscle tissue. Significant age -related differences were identified in principal 3D strain (E) and strain rate (SR) indices (from sequential 2D slices) including in shear

strain in the medial gastrocnemius (MG) and in the soleus (SOL). The tensor projections in the muscle fiber direction and cross-section ( Figure 1 ) did not show any age-related differences but several principal and fiberaligned strain indices showed correlation with collagen 3 from histology. The second part of the study establishes the feasibility of 3D volume acquisition with 3 directional velocity-encoding for calf muscle dynamic imaging to monitor sub-maximal isometric contractions using a 4D Compressed Sensing Flow imaging sequence originally developed for blood flow imaging (Figure 2 ). Skeletal muscles consist of heterogeneous collections of fibres with different metabolic profiles. Due to the different neuronal innervation and fibre composition, each muscle fulfils specific functions and reacts differently to stimuli and pertubations. So far, myosin heavy chain molecules (MyHC) are currently the best marker proteins for characterization of individual fiber types, and several proteome profiling studies have helped to dissect the molecular signature of whole muscles and individual fibers. To precisely quantify fiber typespecific protein patterns, we developed a mass spectrometric workflow to measure skeletal muscle fiber type-specific proteomes. To bypass the limited protein levels in individual fibres, we have designed a proteomics high-throughput fibre typing method (ProFiT) that enables MyHC profiling in individual fibres. Aliquots of protein extracts from separated muscle fibers were subjected to capillary LC-MS gradients to profile MyHC isoforms in a 96-well format. Muscle fibers with the same MyHC protein expression were pooled and subjected to proteomic, pulsed-SILAC, and phosphoproteomic analysis. Our fiber type-specific quantitative proteome analysis confirmed the distribution of fiber types in the soleus muscle, substantiates metabolic adaptions in oxidative and glycolytic fibers, and highlighted significant differences between the proteomes of type IIb fibers from different muscle groups, including a differential expression of desmin and actinin-3. A detailed map of Lys-6 incorporation rates in muscle fibres revealed protein turnover dependent on fibre type, and labelling of mitochondrial respiratory chain complexes revealed a wide range of Lys-6 incorporation rates dependent on the localisation of subunits within different respiratory complexes. Overall, the ProFiT approach provides a versatile tool to rapidly characterize muscle fibers and obtain fiber-specific proteomes for different muscle groups Myonuclei transcriptionally regulate muscle fibres and are key regulators of muscle plasticity. Their cellular location and quantity are important when characterising phenotypes of myopathies, the effect of treatments and to understand the roles of satellite cells in muscle adaptation and muscle 'memory'. 1 We aimed to determine whether PCM1 is, as claimed, 2 a reliable specific marker of myonuclei in-vitro and in-vivo. Single isolated myofibres and transverse sections from mouse, rat and humans were studied from several models including Wild-type and Lamin A/C mutant mice after functional overload, 3 resistance training in rats, 4 and damage and recovery in humans following forced eccentric contractions. 5 Fibres were immuno-labelled for PCM1, Pax7 and DNA. C2C12 myoblasts and human primary muscle cells were also studied in vitro to investigate changes in PCM1 localisation during differentiation and fusion into myotubes. PCM1 labelled the nuclear envelope of myonuclei in mature myofibres and in newly formed myotubes but also labelled centrosomes in proliferating myogenic precursors which may or may not fuse to join the myofibre syncytium. It also labelled non-myogenic nuclei near the sarcolemma especially in regenerating areas of the LMNA+/ΔK32 mouse and damaged human muscle. Such nuclei were found in satellite cells, macrophages, and other interstitial cells. PCM1 is therefore not completely specific to myonuclei, and especially in damaged or regenerating muscle the impact of false-positive identification of interstitial cells on myonuclei counts would tend to cause overestimation of myonuclei per fiber compared with classical counting based on myonuclear positioning relative to the sarcolemma. PCM1 may further prove useful as a marker of satellite cell dynamics due to the distinct change in localisation during differentiation, revealing satellite cells in their quiescent (PCM1-), proliferating (PCM1+ centrosome), and pre-fusion states (PCM1+ nuclear envelope).

Key Words: PCM1; hypertrophy; regeneration; myonuclei. The HERG K + channel is upregulated in skeletal muscle atrophying in response to disuse 1 , cancer 1 , and denervation 2 . Using C2C12 myotubes transduced with HERG-encoded virus, we have shown that overexpression of the HERG channel increases basal intracellular calcium concentration and calpain activity 3 . However, the mechanism by which HERG modulates intracellular calcium levels is not known. To explore this further, we increased intracellular calcium levels by depolarization with 100 mM KCl and used Fura2 dyes and immunoblot to show that HERG does not alter myotube calcium levels by affecting L-type calcium channels (i.e., Cav1.1). Instead, using the SERCA blocking agent thapsigargin with our Fura2 assay, we discovered that the HERG-mediated increase in calcium occurs through modulation of intracellular calcium stores. 4 Therefore, we hypothesized that HERG may be modulating the phospholipase C (PLC)-PIP2-IP3

pathway. To investigate this, we produced an increase in intracellular calcium using bethanechol to activate muscarinic receptors, which activate the PLC-PIP2-IP3 pathway. Using single cell calcium imaging, we reveal that, relative to myotubes transduced with an appropriate control virus, bethanechol treatment of HERGexpressing myotubes produces a significant 2-fold increase (p<0.05) in intracellular calcium levels. The data suggest that HERG may indeed modulate the PLC-PIP2-IP3 pathway. Interestingly, however, this HERGmediated increase in bethanechol-induced calcium levels is not a result of increased PLC activity as demonstrated by IP1-Gq HRTF kit assays, which show no increase in the concentration of IP1 (an IP3 degradation product) in HERG-expressing myotubes relative to controls. However, single cell calcium imaging reveals that the HERG-mediated increase in calcium is inhibited by treatment with the IP3 receptor antagonist Xestospongin-C. The data suggest that HERG enhances calcium release through IP3 receptor activation without affecting PLC activity.

Key Words: HERG; IP3 signaling; intracellular calcium; myotubes. Calcium signaling is the most important pathway which regulates muscle contraction and relaxation. The knowledge of its spatial and temporal distribution among the major compartments in the muscle cells, namely cytosol, sarcoplasmic reticulum, but also mitochondria and T tubular extracellular space, is crucial to drive our understanding in physiological and pathological behavior in muscle mechanics. Despite a general agreement in the fluxes of calcium in skeletal muscle at the qualitative level, there are several open questions in the literature regarding a quantitative assessment of its concentration both in average and temporally/spatially resolved. We approach this problem through a three-dimensional diffusional model of calcium fluxes inside a half sarcomere, based on experimental data on FDB in WT mice. 1 We first estimate the citosolic [Ca2+] comparing the signal of our Fura-2 indicator during a twitch and during a 60Hz stimulation, accounting for the kinetic term through the Schneider equation, 2 with the simulated ratio of occupation of the two Ca-binding sites of the Troponin in the model, and the experimental tension developed by intact muscle fibers. Based on this data, we used the model to assess the relative amount of calcium which enter the mitochondria, which enter from T tubules through SOCE and which exit from the cell through PMCA and NCX. We extended our model to study the calcium fluxes in the parvalbumin knock-out mice, considering two different sets of experimental data recently published. 3, 4 In particular we estimated the possible fate of the calcium released from the sarcoplasmic reticulum in the absence of the major cytosolic buffer and showed the contribution of the export to extracellular space and the uptake into mitochondrial matrix to stabilize the cytosolic concentration.

Key Words: Cytosolic calcium; intracellular signal; local and average concentrations; variations in relation to release from SR. mood and affective symptoms like apathy, anhedonia, anxiet and depression, cognitive dysfunction, and psychotic symptoms such as hallucinosis and delusions. In this context, gait analysis allows to detect quantitative gait variables able to distinguish patients affected by nonmotor mental symptoms from patients without these symptoms. In this research, 68 PD patients were grouped in subjects with and without non-motor mental symptoms based on MDS-UPDRS I (item 1-6). All patients performed gait analysis according to the Davis protocol, 2 both in single and dual-task. Spatial and temporal parameters describing gait were extracted, as already done in previous researches. 3 The results of statistical analysis showed significant different gait patterns in patients with non-motor mental symptoms as compared with those without. Single-task variables showed that 9 out of 16 spatiotemporal features were statistically significant for the univariate statistical analysis (p< 0.05). In addition, a statistically significant difference was found in stance phase (p=0.032), swing phase (p=0.042) and cycle length (p=0.03) of the dual task. In conclusion, PD patients with non-motor mental symptoms versus PD patients without this cluster of symptoms showed a worse gait pattern in both single and dual task, thus supporting the notion of a more widespread disease involving neurotransmitters systems other than dopamine in patients with neuropsychiatric symptoms.

Key Words: Parkinson's Disease, non-motor mental symptoms, gait analysis Knee Osteoarthritis is one of the most common form of arthritis and affects a significant percentage of the world population. 1 Multiple indexes created to classify the cartilage status are present in the medical literature and are mainly based on physicians' image analysis and subjected to different and personal interpretations. 2, 3 In the frame of the European Project RESTORE a database of MRI and CT-scan images of degenerative, traumatic, and healthy patients is developed. Approximately 100 objective features are extracted from both 2D, and 3D In this research a novel and unique Motion Sickness moving platform is used to study different postural control reactions. 1, 2 The platform simulates a rough sea environment through virtual reality which visualize waves that coordinate the platform movements. Together with EEG, EMG, and hearth rate sensors a force platform is installed under the subject feet during the experiment. This research is focused on the extraction of multiple features from the force platform to study the postural movements of the different subjects before, during and after the sea simulation. These features are then selected using a machine learning approach focusing on the ones which allows to distinguish between motion sick and not sick subjects with higher accuracy. Then the different postural behaviors during the simulation are confronted between different classes of participants like elderly and young, sporty, and not and others. Finally, the selected features are correlated with the EMG data to study the relations between postural and muscle reaction to the induced motion sickness environment. Total hip arthroplasty (THA) is the most widely used surgery in case of severe damage of hip joints. However, the implant produces significant variations of stress distribution in the operated femur, determining the host bone remodeling, which, in turn, influences the bone quality of THA patient. The aim of the study is to create a prediction model, which allows determining significant parameters for THA femur quality during the remodeling, starting from patient clinical data, diagnostic x-ray computed tomography (XCT) and 3D high resolution imaging. Patient clinical data as age, gender, body mass index, type of operation (unilateral or bilateral), prosthesis (cemented or uncemented) and

diagnosed pathologies connected to bone tissue, are correlated to multi-scale parameters of femur. Information at macro level is obtained by the analysis of diagnostic X-ray computed tomography (d-XCT) scans of patient femurs. Data are segmented to collect the densitometry of regions of orthopedic interest (gruen zones). The densitometry of rectus femoris muscle is also considered, due to the fact that remains roughly recognizable also when it is degenerated. Micro and nano femur morphometric parameters are obtained by high resolution 3D imaging on femur biopsies, achieving information on bone microarchitecture and ultrastructure.

After a learning, based on above-mentioned THA patients' data at 24 hours, 1 years and 6 years after the implant, the prediction model can predict the bone quality and the better-regenerated zones. The prediction model of THA femur quality works also as support of further clinical decisions on post-surgery treatments and rehabilitation, by providing a projection on patient mobility. Neck pain is an increasing healthcare problem throughout the developed world and is the fourth leading cause of disability globally. It can be difficult to diagnose and treat effectively because of the complex pathophysiology and variable presentation between patients. Misdiagnosed and improperly treated neck pain can develop into chronic conditions that last for years after initial onset. Despite a rise in the incidence of neck pain (21% globally between 2005 and 2015), 1 there is no standardized methodology to objectively define the underlying pathology that is the cause of the pain. The Icelandic startup company, NeckCare, had developed a system, measuring the kinematic variables of the head and neck. 2 The three main pillars of clinical assessment are: i) Proprioception: Measures the deviation in degrees from the initial neutral position of the head after a blindfolded subject has rotated the head to the left/right/flexion/extension and back to find again the initial position of the head; ii) Neuromuscular control (The Butterfly test): Measures the movement control of the head. The subject controls a cross on the screen and the objective is to chase a moving target. The outcome measures are average distance away from the target, the percentage of time spent on the target and the smoothness of the movement. The Butterfly trajectories are divided into three difficulty levels that come up in random order and the patient will only see a single point moving and not the overall trajectory. The trajectories along the measurements for the two groups can be seen in Fig. 1 ; iii) Range of motion: Measures the overall range of motion in each plane. The outcome measures are in degrees. Measurements were taken of a control group and patients suffering from whiplash associated disorders (WAD). The measurements are carried out so that the subject was sitting in a chair 90cm in front a computer monitor that was adjusted so that the middle of the screen was at eye level. A movement sensor was placed on the head that connected via Bluetooth to a computer. The movement sensor used an IMU sensor which calculates the 3D Euler angles of the head and projects them onto the monitor for a visual feedback. The results demonstrated that there was a significant difference between the neuromuscular control of the control group and the whiplash group in the Butterfly test. The whiplash group was on average further away from the moving target and spent less time in the vicinity of it than the control group. The whiplash group had less range of motion in all planes and poorer proprioceptive control. These tests can help to identify the muscular impairment for patients suffering from whiplash associated disorders and can be used in the rehabilitation of these patients to than treatment. This leads to large variability and confounds the observed outcomes: "Garbage In, Garbage Out", whereby the data quality determines the quality of the results. Therefore, it is becoming increasingly important to find the right input and critical success factors to describe medical quality outcomes in musculoskeletal rehabilitation, where translational myology can make important contributions. We present clinical results for relevant moderating variables from rehabilitation practice and the performance score stratification (T2D). Associations between clinicianreported and patient-reported outcome measures are low to moderate. Overall, musculoskeletal rehabilitation in pre-post designs leads to an outcome with a large effect size. Women, the elderly, and patients with obesity and low physical activity often have poorer baseline scores (t1); therefore, they sometimes appear to respond better to interventions in an inpatient setting. Because the improvement depends on the patient's baseline status (t1), the simple formula T2D = t2 + (t2 -t1) better reflects outcomes, taking into account the functional status at the beginning of rehabilitation ( Figure 1 ). We believe that the T2D concept is highly relevant for many studies and allows better clinical discrimination. T2D may be an appropriate external selection and validation criterion for new opportunities and applications, such as artificial intelligence for the diagnosis and management of myopathology. The interest in potential of eccentric training in older adults seems to be increasing in the recent years. Two very recent systematic reviewers focused on the effect of eccentric training in healthy older adults (Kulkarni et that eccentric exercises can be as effective as conventional exercises in older adults for improving functional performance. We performed a broader review and included 18 studies that compared eccentric and concentric exercise interventions. Our analyses suggest that eccentric exercise is actually superior to concentric exercise in terms of the effect on functional performance. Specifically, larger improvements after eccentric compared to concentric exercise were seen for timed up and go test, 2-min stepping test and 30-s sit-stand test (moderate effect sizes). There was also a tendency for eccentric exercise to induce larger improvements in muscle strength and size, but the overall effects were not statistically significant. Together with previous reviews, our analyses suggest that eccentric training may evoke superior effects to conventional/concentric training, wherein functional performance outcomes were most illustrative.

Key Words: Flywheel training; eccentric exercise; elderly, functional outcomes. One of the effects of the global warming is the increase of heatwaves in terms of intensity, frequency, and duration. While global warming is a worldwide concern, heatwaves is a local issue depending on land use change, urbanization, orography, and meteorological conditions. 1,2 Even the local specificity of heatwave, in almost all regions a measure of cumulative heat shows significant increases since the 1950s, mainly driven by heatwave days. 3 Understanding past decades heatwave trends and future projection have strong implications to know the impact on human health, on ecosystems and on infrastructure planning. 4 Here we will show how temperature is changing at different scales from global to local, 5 focusing on the formation and evolution of urban heat islands. The analysis will cover the last 200 years of data and projections of future trends. Finally, will be discuss the impacts of heatwaves on ecosystems and human health pointing out drivers, mitigation, and adaptation strategies.

Key Words: Climate Change, heatwaves, radiative forcing.

Climatic changes are increasing the incidence of heat waves, periods in which the temperature raise above the average temperatures for three or more days. Heat waves are very dangerous for human health, as it is now clear that the mortality rate increases dramatically during those days. 1 Even if several factors may contribute to sudden death in high environmental temperatures, the most common cause of death attributable to heat is dehydration, heat cramps and exhaustion, and hyperthermia, i.e. in one word heat stroke (HS). 2,3 The cellular and molecular mechanisms underlying HS are multiple and still debated; as a result: a) a cure for acute treatment of HS is still missing; b) patients are usually treated only with supportive measures (cooling and hydration); and finally c) clear guidelines for proper lifestyle habits before and during heat waves are not well defined. We have collected compelling evidence that HS shares common symptoms with malignant hyperthermia (MH) susceptibility (Figure) , a life-threatening syndrome caused by mutations in proteins deputed to Ca 2+ handling in muscle and triggered by administration of halogenated anaesthetics. There is general agreement that the main mechanism triggering MH crises is an abnormal SR Ca 2+ leak through mutated RYR1, causing uncontrolled contraction and rhabdomyolysis. However, other factors must be taken in consideration: A) Ca 2+ leak from the SR triggers a feed-forward mechanism leading to overproduction of reactive species of oxygen and nitrogen (ROS and RNS) by mitochondria, which in turns causes further Ca 2+ release through RYR1-nitrosylation. 4 B) MH responses do not occur in the absence of extracellular Ca 2+ (5) suggesting that Ca 2+ influx from the extracellular space is an essential component to be considered. The Central Hypotheses guiding our recent work are: a) skeletal muscle plays a central role in HS; b) lifestyle habits may affect proper muscle function, hence cause either increased or diminished risk of the normal population to HS in challenging environmental conditions.

Heat-stroke (HS) is a life-threatening response to heat characterized by an abnormal increase in body temperature (>40°C) that causes dysfunction of organs, central nervous system and may end in death. 1 Environmental heat-stroke (EHS), often triggered by a hot and humid environment, is caused by excessive heat production in muscle, which in turn is the result of abnormal Ca2+ leak from the sarcoplasmic reticulum (SR) and oxidative stress. 2 See Figure for a Mechanistic Model. As high fat diet is known to increase oxidative stress, 3 the objective of the present study was to investigate the effects of high-fat diet in the heat-stroke susceptibility of c57bl/6 wild type (WT) mice of 4 months of age (adult). Our results show that, in comparison with mice fed with a control diet, mice after 3 months of high-fat: a) increased heat generation and energy expenditure (assessed by indirect calorimetry) during heat stress; b) elevated oxidative stress in both EDL and Soleus muscles; and c) enhanced sensitivity to caffeine and temperature of isolated EDL and Soleus muscles during in vitro contracture test (IVCT, the gold standard procedure to test in-vitro EHS susceptibility). Our data suggest that high-fat diet predispose mice to EHS, possibly as a result of increased oxidative stress and excessive release of Ca2+ from SR. This study may have important implications for guidelines regarding food intake during periods of intense environmental heat.

Key Words: High-fat diet, heat-stroke, oxidative stress.

-86 -CAST, Center for Advanced Studies and Technology; University G D'Annunzio of Chieti-Pescara, Chieti, Italy. Barbara Girolami, Email: barbara.girolami@unich.it Exertional/Environmental Heat Strokes (EHSs) are hyperthermic crises triggered by strenuous physical exercise and/or exposure to environmental heat, which are caused by an altered intracellular Ca 2+ homeostasis in muscle. 1, 2 We recently demonstrated that a single bout of exercise on treadmill leads to formation of Calcium Entry Units (CEUs), intracellular junctions that promote interaction between STIM1 and Orai1, the two proteins that mediate Store-Operated Ca 2+ Entry (SOCE). SOCE is a mechanism that is activated during muscle fatigue and that allows recovery of extracellular Ca 2+ during prolonged activity. 3, 4 The hypothesis underlying this project is that assembly of CEUs during prolonged exercise may predispose to EHS, when exercise is performed in challenging environmental conditions. To test this hypothesis, we used 4 months old WT mice that were: a) first, divided in 3 experimental groups: control, trained-1m (1 month of voluntary running in wheel cages), and exercised-1h (1 hour of incremental treadmill run); and b) second, subjected to an incremental treadmill run of 45 min at 34°C and 40% humidity. We then: i) measured the internal temperature of mice, which was higher in the pre-exercised groups (trained-1m: 38.9°C ± 0.33; exercised-1h: 38.7°C ± 0.40) compared to control (37.9°C ± 0.17); ii) applied an ex-vivo exertional stress protocol to isolated EDL muscles (tetanic stimulation performed at 30°C) and verified that samples from trained-1m and exercised-1h mice generated a tension significantly greater than control; iii) Analyzed CEUs by electron microscopy (EM) and verified that EDL muscles of exercised-1h ad trained-1m mice contained a greater number of elements forming CEUs. The data collected suggest that assembly of Calcium Entry Units during exercise could predispose to EHS when exercise is performed in challenging environmental conditions. Movement is encoded by spinal motor units. The motor unit is formed by a motoneuron, and the muscle fibres innervated by its axon. The motoneuron receives efferent and afferent synaptic inputs that modulate the force output of the muscle. There is very little knowledge on the changes in motor unit behaviour during natural dynamic movements consisting of changes in muscle length and neural drive. Moreover, the associations between kinematics, kinetics and motor unit dynamics are largely unknown. Here we aimed to explore these associations with a linear and nonlinear method. With the linear method, we decomposed high-density EMG signals into constituent motor unit action potentials during dynamic and isometric hand movements. With the nonlinear method, we built a framework with digital cameras that recorded the movements of the hand as well as recording isometric forces with an instrumented circular object. Deep neural networks were trained to understand the exact position of the hand from the digital cameras as well as from the high-density EMG signals. The movements consisted of more than 20 degrees of freedom, ranging from individual and combined finger flexion/extensions, and different hand and wrist gestures at different speeds (0.5 and 1.5 Hz). Four hundred EMG channels recorded the activity of the forearm muscles. The non-linear method was able to identify with 99.6% accuracy the exact movement of the fingers. The decomposition of the electromyogram revealed unique motor unit firing patterns that were significantly different than the control during isometric contractions. The delays between the onset of neural activity and movement kinematics were similar across all fingers and decreased with the speed of force contraction suggesting that the central nervous system compensates the muscle dynamics with unique control strategies for each finger. Because the hand digits were flexed and extended at various speeds, our interface shows that the dynamical output of the hand is fully explained by the neural strategies and that the musculotendinous unit has only a passive role for the rate of speed production. Successful performance of motor tasks requires the adequate modulation of force output, which depends, in part, on the integration of information from various mechanoreceptors located in muscles, muscle-tendon interfaces, and joints. These signals form the basis for the perception of self-movement and body position -termed proprioception -which is defined as the conscious or unconscious perception of the position of different parts of the body in space. Along with the kinaesthetic sense (the senses of position and movement), the sense of force is another aspect of proprioception. The sense of force represents the ability to correctly perceive and reproduce a given level of force. However, it is still debated whether the sense of force really relies on proprioceptive inputs. This may depend on the experimental approach used for its assessment. Indeed, it can be evaluated by asking the participants to match with one limb the force produced by the contralateral limb (two-arm matching task), or by asking to reproduce a force with the same limb (force reproduction task). It has been suggested that the twoarm matching task does not require proprioceptive feedback but rather a centrally generated signal. In contrast, the force reproduction task is suggested to mainly rely on proprioceptive inputs, including signals from muscle spindles and Golgi tendon organs, in addition to sensory inputs from skin mechanoreceptors. By manipulating proprioceptive and visual inputs within different phases of the force reproduction task, we provide relevant elements supporting the role of proprioceptive signal from muscle and tendons sensors during the force reproduction task. Furthermore, we show a decrease in the sense of force with ageing, which could reflect age-related decline in sensorimotor integration of proprioceptive and somaesthetic inputs. Age-related changes in the neuromuscular system functions may affect profoundly high-level athletes' performance across their career, and potentially determining its length. Therefore, the present study aimed to explore the role of age on neuromuscular functions in a group of competitive athletes, and in particular, to analyse the fatiguing effect of a maximal intensity sprint session (MISS). Thirty-one competitive endurance athletes completed a knee extensors and flexors' maximal-voluntary-isometric-contraction (MVC) test (3x5s, 60s recovery) before and after a maximal intensity sprint session (MISS) consisting of 4x15s Wingate-tests. The data have been stratified considering three age categories (18-28, n=11, 29-38; n=10; 39-43, n=10). Non-significant differences in neuromuscular performance markers emerged between the three age groups at baseline (p>.05). Overall, both quadricep and hamstring muscles early and late RTD dropped significantly more than the MVT (p<.05). Age had a significant effect on early RTD with older athletes exhibiting greater RTD drop compared to younger athletes (p<.05). A significant effect of age emerged also for the changes in surface electromyography (sEMG) variables, in which the frequency spectrum variables (MDF and MNF) dropped significantly more than the sEMG amplitude (RMS) (p<.05). In addition, age was largely significantly correlated with early RTD drop. On the contrary, no significant effects of age emerged for the MVT and late RTD drop (p>.05). The dynamics of changes in neuromuscular performance markers after a MISS suggested that getting older, competitive athletes may potentially experience greater lost in early explosive strength compared to maximal or late explosive strength, due to neuromuscular fatigue. After 6 month 68% of COVID-19 survivors, and still 49% after 12 months, show one or more long-COVID symptoms. 1 The symptoms persist regardless of COVID-19 severity. 2 The public attention to Long-COVID contributed to shed light on a similar but neglected syndrome. Chronic fatigue syndromes after acute infections (e.g., Ebstein-Barr virus) are known since decades, but mostly are without adequate recognition from medical professionals. 3 Based on established diagnostics, there is still no satisfying approach to understand the phenomena of post-infectious syndromes and, consequently, to apply purposeful therapies. In view of the immense and increasing number of Long-COVID patients, diagnostical approaches are urgently needed. The most common symptoms of post-infectious diseases are fatigue and muscle weakness. 1 In clinical practice, a considerably reduced ability to resist external forces is regularly seen in those patients. Therefore, it is questioned whether the newly inaugurated Adaptive Force (AF) could be a promising approach to measure post-infectious states. The AF reflects the capacity of the neuromuscular system to adapt adequately to external increasing forces in an isometric holding manner. Healthy persons show a high maximal isometric AF (AFisomax (N)) assessed during manual muscle test (MMT) objectified by a handheld device. 3, 4 Thereby AFisomax is close to the maximal AF (AFmax (N)) reached in the same measurement, reflecting a proper adaptability of the neuromuscular system. AF pilot measurements of Long-COVID patients gathered in clinical routine (no clinical study) will be presented. The AF of the hip and elbow flexors of one side was assessed manually (MMT) each three times (alternating) by wellskilled examiners objectified by the handheld device. The maximal holding capacity (AFisomax) and the total maximal AF (AFmax) were evaluated (Fig. 1) as well as the ratio of AFisomax/AFmax (%). Long-COVID patients show a substantial and significantly lower AFisomax and AFisomax/AFmax compared to healthy subjects. After successful treatment the holding capacity is immediately improved and the ratio is significantly higher compared to the initial measurements. This suggests the holding capacity is essentially lower in postinfectious conditions and could possibly be a sensitive biomarker for post-infection fatigue. These preliminary data need to be confirmed by future clinical studies. In case of verification it will be used to optimize individual diagnostics and to apply purposeful treatments.

Key Words: Adaptive Force; holding isometric muscle function; long-COVID; post-COVID. Electrical stimuli play an important role in many clinical diagnostic and therapeutic methods as well as in experimental and clinical studies. Key for understanding elicited physiological mechanisms is understanding of interaction of the induced artificial electrical fields with electrically excitable neural and muscular structures. Adjustable variables in stimulation setups are size and position of electrodes, 1 shape of stimuli, sequence of their delivery and variation of their size and rate. Goals are selective activation of neurons or, in case those are denervated, muscle fibers, and safe delivery of currents, without harm to skin and other tissues. Coupling of the technical systems to the physiological structures is made via contacting electrode surface, at the skin or embedded in soft tissue, with a transition from a technical electron conductor to a physiological ion conductor. Careful provisions are necessary for keeping associated electrolytic processes safe and avoiding tissue damage risks, acutely and in long-term perspective. Functional coupling happens at the electrically excitable membranes, which form the envelope of nerve or muscle -90 -fibers. At such membranes, induced electrical fields can elicit traveling action potentials, starting at a trigger point, where a discharge condition is reached, in addition, there are also control options, where the induced field influences resting potentials and increases or lowers excitability for reaching a discharge threshold. 2 Basic principles of fiber recruitment are the size dependence, the bigger the fiber in diameter, the lower the discharge threshold, and distance dependent, threshold increases with distance between electrode and neuron. 3, 4 In the mixed peripheral nerve, preferred recruitment of groups of large proprioceptive afferent neurons is reached with lowest stimulation intensity, followed by co-recruitment of cutaneous afferents, efferent neurons of type 2 motor units and then type 1 motor units, and finally smaller afferent pain fibers, when stimulation intensity is increased (Fig. 1) . It is important to consider this as limitation, when physiological responses are interpreted, as recruitment has strong impact in central interneuron processing of afferent input signals as well as in motor control and metabolic processes in activated muscles. In addition to stimulus size dependent recruitment, frequency of trains of stimuli plays a major role in both central neural processing and peripheral activation of muscles 4 .

Key Words: Functional Electrical Stimulation, neuron recruitment, fiber size, threshold project (RISE) has demonstrated that a stimulation period of two years led to significant changes in muscle growth and reduction in adipose tissue in the lower limb muscles. 4 It is still unclear after what time the transformation process from connective and fatty tissue to contractile muscle tissue begins in chronic paraplegia with damage to the lower motor neuron. Likewise, the process of this conversion and how it occurs remains unclear. In other words, is there an immediate visible increase in muscle mass or does connective and adipose tissue break down first and when do these changes in tissue composition start to occur? Kern and Carraro have visualized and emphasized on the importance of life-long stimulation in people with lower motor neuron lesion in order to maintain proper blood circulation and to produce a so-called "cushioning effect" in people with spinal cord injury (SCI) to prevent them from skin injuries. 5 The aim of the present study is to investigate the effect of electrical stimulation of chronic denervated gluteal muscles in the first six months after stimulation onset. Changes in muscle as well as adipose tissue thickness were assessed over the stimulation period of six months using standardized magnetic resonance imaging (MRI) measurements of the gluteal area on the projected line of the sacroiliac joint (SIJ), the acetabulum and the coccyx. The study participants (four participants with AIS A and one with AIS C) stimulated the denervated gluteal muscles five times a week during a period of six months. The stimulation was composed of a warm-up phase and a training phase lasting a total of 33 minutes per session. The electrical stimulation was administered with the Stimulette den2x (Schuhfried GmbH Vienna, Austria) with the sponge electrodes being placed on the buttocks to generate a horizontally oriented electrical field to cover the entire gluteal area. Key Words: Denervation, Electrical stimulation, SCI, Tissue composition, Magnetic resonance imaging.

Epidural stimulation of the spinal cord (eSCS) has for several decades been used successfully for the treatment of pain and spasticity. It has also been used to facilitate voluntary control of walking by individuals with partial lesion. Newly (Feb. 2022) walking in complete paraplegic individuals has been demonstrated using eSCS. eSCS has the advantage compared to tSCS that the electrodes are placed near to the stimulation target and can activate these with less coactivation of other muscles. This though given that the placement of the electrodes can be done during an operation and that they are stable in place. Using leads with an electrode array makes it possible to adjust for misplacement and capsuling through connective tissue to some limit. tSCS has, following the experience with eSCS, been used with similar results for spasticity treatment and facilitation of voluntary controlled walking in spinal cord injured (SCI) individuals. This is achieved using common gel electrodes on the surface of the skin above T11 -T12. Being further away from the targeted cells the specificity of tSCS through localisation of the current field is lower than by eSCS but sensory fibres in the posterior roots have a larger diameter than lower motor neuron fibres and can be targeted with lower current intensity without activating the latter. In resent years progress has been made in evaluating spasticity although there is a lack of consensus in the definition of spasticity. Also, our group has developed a new parameter for quantifying spasticity based on Wartenberg pendulum test targeting the excitability of the stretch reflex. It is defined as the time span from 50% of the maximum velocity of the first leg swing, determined from the pendulum test, to the activation of the muscle contraction in the quadriceps, determined from the EMG. The result is a parameter reflecting changes in excitability of the stretch reflex. Spinal cord stimulation effect on the brains or the spine plasticity has not been in focus of research so far. Our group has developed a methodology to evaluate EEG, EMG and joint angle measurements made during several common neurological and physio therapeutical tests as well as SCS in order to analyse the reaction of brain and muscles to these tests. Pipeline for brain network analysis and time-frequency analysis of the neurological signals has been developed. This enables studies of changes in the brain due to different condition of the subject. Epidural spinal cord stimulation (eSCS) has shown to be an effective way for neuromodulation in the spinal cord. Treatment of pain with eSCS has a long time been a clinical routine. Also, has the treatment of spasticity in SCI been proven to be effective although it has not found its way into a daily clinical application, mostly due to the fact that it is an invasive treatment. In recent years several groups, including ours, have shown that tSCS can be used to treat spasticity and facilitate walking in SCI in a similar way as eSCS. This is not the case for brain insulted individuals. It is believed that the tSCS affects the spinal cord neural network with inhibiting signals partly replacing the brain's function. This process is poorly understood. Also, the interaction of the spinal cord with the brain or the effect of tSCS on the spine and brain has not been the focus of research. Plastic processes in the spinal cord and in the brain as a result of SCI are known to some extent but not the effect of tSCS on the same organs. The neural structures influencing the activity on the lower motor neuron are the target of the treatment of spasticity in the lower extremities. These effects and the structures are to be studied in this work through the monosynaptic reflex and tSCS effects mainly through EMG of the thigh and the connection to the brain through EEG. In this project a measurement system for recording of H-reflex and m-wave as well as EEG and joint angles during patellar reflex test and tSCS is constructed and tested. Especially exact time measurements (Δt ≤ 0,5 ms) between events are targeted. Additionally, EEG, EMG and joint angles registrations during three standard tests are evaluated. They are 10 m walk, stand up and go and five times stand up (if movement ability is there). This is done by healthy, SCI and brain insulted individuals. These tests are standard for evaluating movement ability in individuals with lesions in the central nervous system (CNS). A tested and evaluated measurement system enabling exact time measurements between events for research on the monosynaptic reflex arc including H-reflex and m-wave. Maps and statistics on individual variability between healthy subjects as well as SCI and brain insulted. Also, a tested and evaluated measurement system for research on EEG, EMG, and joint angle signals. Recordings, processing, and evaluation of brain potentials evoked by patellar tendon test and tSCS and statistical analysis on brain activity between groups. Skeletal muscle (SKM) loss is considered to be largely irreversible and therapeutic options to restore muscle mass have been limited. Frequent reasons for SKM loss are injuries, surgery, aging, metabolic diseases, as diabetes, and inherited genetic diseases. SKM has good regenerative capacity due to the presence of resident stem cells, the satellite cells, located between the cell membrane and the basal lamina, however in vitro studies demonstrated their poor ability to migrate and to selfrenewal in situ: that means their intrinsic proliferative capacity is lost in normal or suitable cell culture. New strategies are required to overcome these limitations; recently, various studies have focused on recreating the Models adequately mimicking skeletal muscle injuries and diseases are important tools to understand and study mechanisms underlying initiation and progression of diseases as well as tissue regeneration and remodeling after injuries. 1 3D tissue models help increase significance of generated results due to their similarity to in vivo conditions compared to conventional cell culture without the obvious drawbacks of animal tests. Herein, we present how an established tissue engineering approach to create biomimetic skeletal muscle-like tissue can be used to model pathologies. Healthy tissue constructs created through mechanical stimulation of murine myoblasts with a bioreactor, 2 and subsequently subjected to different types of stress present the basis of this study. Stress was induced either by 1.) mechanical overstimulation with high frequency or high tensile stress with the same bioreactor; 2 2.) complete deprivation of mechanical stimulation or 3.) by exposure to oxidative stress with another bioreactor system based on reactive oxygen species generated through hydrostatic pressure. 3 Mechanical overstimulation resulted in an immediate stress response (Fig. 1 ) as well as prolonged downstream signaling activity. Furthermore, high frequency overstimulation led to increased gene expression of markers of proliferation and early-and mid-stage myogenesis, and changed morphology of myotubes (Fig.  2) . Deprivation of mechanical stimulation for seven days led to severely impaired integrity of myotubes on a morphological level (Fig. 2) , but also to downregulation of signaling pathways involved in stress response and myogenic development. Oxidative stress induced decreased viability (Fig. 3) and loss of structural integrity of myotubes. Thus, we conclude that the first steps towards the establishment of a biomimetic tissueengineered model for skeletal muscle pathologies were successful. Nevertheless, further refinement and investigation of the mechanisms involved are still needed. 

Oxidative stress with the help of drug therapy to improve and normalize metabolic processes, as well as the development of an adaptation system for the changes caused by the pathological process. It is important to take into account theoretical aspects of the organization of physiological and pathological movements, as well as the formation of short-and long-term compensatory processes of damaged structures and the impaired functions of the body systems in the course of neuro-rehabilitation. The formation of compensatory mechanisms is greatly influenced by signaling from the external and internal environment of the body, i.e. feedback. 1 A reliable brain function is ensured by dynamic inter-neuronal activity, resulting in morphologically redundant connections. Neuro-rehabilitation is also based on the systemic organization of the brain functions, the reserve capacity of the brain in reorganizing its structures under the conditions of the pathological process. This reorganization causes the patient's motor retraining, which leads to the restoration or compensation of the impaired functions. When conducting neurorehabilitation, it is necessary to strive for a true restoration of the disturbed functions of the patient's body. This level of HP seems to be the highest and lies in the fact that the functions of the body completely return or are as close as possible to the original state. The main mechanism ensuring a true functional recovery is the disinhibition of inactivated nerve elements by means of techniques aimed at stimulating them. 2 With brain damage, recovery occurs due to intact functional systems. This process is ensured by the plasticity of the nervous system and the anatomical connections between its departments. The plasticity of the brain lies in the ability of the nervous tissue to change its structural and functional organization under the influence of external and internal factors. It is due to the following mechanisms: the functioning of the previously inactive connections; sprouting of the fibers of the remaining neurons (renewed of growth of the affected axon, changes in fiber branching, the area and density of the dendritic spines); formation of new synapses; synaptic remodeling (changes in the configuration and properties of synapses); reorganization of neuronal circuits; extrasynaptic transmission of excitation; a change in the structure of astrocytes, an increase in the number of contacts between synapses and astrocytes. During the medical and social rehabilitation, one must bear in mind that severe disabling neurological diseases cause permanent disability and social maladjustment. Thus, regarding the main aspects and principles of neurorehabilitation, it is important to understand the complexity, aims and problem orientation of this process, which is primarily aimed at increasing the degree of recovery of neurological functions of patients after a stroke, their level of social adaptation and, ultimately, improving the quality of life of patients and their relatives. 3 

Words: Neuro-rehabilitation; stroke; neuroplasticity.

possibility of measuring only ONE or more aspects, and not the whole complexity. The same stimulus can in fact elicit different responses in the same individual in relation to time and environment. It is essential to associate the measurement of the phenomenon with a series of numbers that allow its interpretation, temporal evolution and comparison with other phenomena. In the scientific field, the numerical description of biological functions needs reliable instruments that provide accurate and repeatable values of one or more characterizing parameters. The response to the stimulus is not solely determined: the input-output relationship can lead to the same result by exploiting different paths. Hence, despite the knowledge of the input and the output, the biological phenomenon is not necessarily known. Each individual owns a specific adaptability to the different stimuli that undergoes fluctuations over time. When an input exceeds the adaptability threshold, unexpected and previously undetectable problems may be generated. In the clinical dental field, it is necessary to quantify the function and/ or dysfunction and the need of intervention to restore the functional alteration, by means of objective measures and not only of patient subjective symptoms. It is crucial to set instrumental tests with dedicated protocols that allow to measure the stability of the occlusal relationships both in static and dynamic conditions and to uniquely study the experimentally-induced variations. The evaluation should integrate the functional aspects with the morphological ones at micro and macrostructural levels. The research activity of the Functional Anatomy Laboratory of the stomatognathic apparatus (Functional Anatomy Research Center-FARC-University of Milan) has been developed aiming at defining standardized indices that measure the overall activity of the chewing muscles. Strength, recruitment and coordination of muscles are gathered in order to identify the impact of various occlusal conditions on the masticatory synergism and possible effects on other districts, primarily on the neck. The FARC 30-years-experience applied innovative non-invasive technologies (electro magnetic and electromechanical digitizers, optical morphometric methods, surface electromyographs, dynamometers, etc.) for the creation of protocols that clinician could use to measure the chewing function in each individual patient in a simple way, testing its fluctuations over time.

Key Words: Physiological functions; scientific instrumentation; biological measures; masticatory muscles activity.

The purpose of orthodontic treatment is to guide the development of the upper and lower jaw during childhood and adolescence to establish proper eruption and arrangement of teeth in the alveolar bone in balance with oral functions. 1 Centripetal forces are exerted on the dental arches by the perioral muscles and centrifugal forces by the tongue. These structures exert slight pressures at rest for prolonged periods of time and are therefore fundamental in determining the dental position. Alteration in these forces influence the orientation of the skeletal structures thus resulting in malocclusion. 2 The development of the masticatory system is mostly determined by hereditary and environmental factors. In this context the activity of masticatory and perioral muscles is determinant not only in the setting of malocclusion but also in the definition of the treatment plan and in the long-term retention phase. 3 During childhood the highest risk to develop malocclusion is linked to alterations in the normal pattern of oral functions (atypical swallowing, oral breathing, bad oral -97 -habits) with a consequent modification of the skeletal relations between the two jaws. In this period orthopedic and myofunctional treatment are pursued to restore a correct balance in the jaw's growth. 4 In adolescence patients presenting skeletal class II malocclusion (most frequent malocclusion in Europe) are mostly treated by using functional appliance to correct the discrepancy between the upper and lower jaw (increased overjet). To do so the removable appliance stress out the soft tissue and muscles fibers to allow a forward repositioning of the mandible. 5 In adults' patients the lack of residual growth affects the treatment toward a surgical procedure or a camouflage treatment. Even in this period, muscles activity represents a determinant element in the treatment planning and in the long-term stability. 6 Key Words: Orthodontics; malocclusion; muscles activity; myofunctional therapy.

Dental occlusion changes may be needed for several reasons. Osseous and dental static and dynamic relationships alterations often result from severe tooth wear, crowding, loss of masticatory units and oncologic surgical resections that require reconstruction of dental and/or skeletal occlusal relationships. Any dental work that changes the occlusal surfaces and/or the tooth position could potentially modify oral proprioception. Kogawa., et al. 1 demonstrate how the interdental perception threshold in healthy subjects is less than two hundredths of a millimeter. Neuromuscular system could be forced to develop functional adaptations to dental treatments involving occlusal surfaces that change oral proprioception. It has been clearly demonstrated how dental afferents play a role in the masticatory muscles recruitment. Occlusal modifications could change masticatory muscles global contraction intensity but also their functional relationship. 2 Moreover muscle imbalances may originate from several factors including interference in the working or balancing side and loss of posterior vertical support. 3 Indeed, it has been demonstrated how proprioceptive dental alterations due to iatrogenic occlusal disturbances (200 μm thick) may require also neck muscles functional adaptations. 4 The relationship between abnormal muscle recruitment and symptoms such as pain or signs of dysfunction like movement limitation does not appear linear, underlining that a great number of subjects has a wide functional adaptability. 5 The absence of clinical symptoms (mainly pain) following an occlusal intervention does not correspond directly to a procedure free from imperfections and/or anomalies. The nervous tissue can adapt the muscles rectruitment to new oral conditions (without causing symptoms such as pain), 6 "masking" changes in other structures, for example, teeth, bones and joints. Occlusal proprioception requiring (asymptomatic) muscle adaptations could causes changes in the distribution of the occlusal forces causing the following major complications: 1) Functional. The adaptation capability is not constant between individuals (some patients may develop symptoms as a result of altered occlusion) and over time; 2) Biological. Mechanical forces play a role on bone biology; the bone apposition and resorption mechanisms are managed by chemical processes initiated by mechanical stimuli. It has been demonstrated how the bone modifies in order to support functional loading needs; 3) Mechanical. The prosthetic (crowns and implants) reliability is the result of the stress developed in artificial products. In this context the use of masticatory function instrumental evaluations (before and after therapies that modify dental occlusion) are suggested to support the clinician to quantify the effect of occlusal changes on the oral biology. In fact, the biomechanical studies show that each muscle has its own specific action vectors (or more than one) and that changes in the muscle forces distribution cause alterations of the mechanical stresses on the hard structures. 7 In conclusion, a reliable dental treatment include masticatory functional evaluation to ensure greater reliability and durability of homogeneously stressed oral rehabilitations. Osteoarthritis (OA) is a degenerative pathology of the joint, 1 characterized by progressive damage to articular cartilage, which currently represents an important cause of disability, 2 with an estimated prevalence ranging from 1 to 10% in the general population. 3, 4 Despite several mechanisms having been proposed to explain its onset and progression, the aetiopathogenesis of osteoarthritis is still doubtful. This study hypothesizes an association between fascia lata alterations and hip OA; in particular, changes in the production of collagen and HA could result in altered fascial structure and behaviour in OA patients. The structure and composition of fascia lata in healthy subjects and in patients with hip OA were compared with the aim to evaluate any differences in the amount of Collagen type I (COL I), Collagen type III (COL III), and Hyaluronan (HA). Ten samples from healthy subjects and 11 samples from OA patients were collected. COL I was significantly more abundant in the OA group (p = 0.0015), with a median percentage positivity of 75.2 % (IQR 13.11), while representing only 67 % (IQR: 8.71) in control cases. COL III, with median values of 9.5 % (IQR 3.63) (OA group) and 17.10 % (IQR 11) (control cases), respectively, showed significant reduction in OA patients (p = 0.002). HA showed a median value of 10.01 % (IQR 8.11) in OA patients, denoting significant decrease (p < 0.0001) with respect to the control group median of 39.31 % (IQR 5.62). The observed differences suggest a relationship between fascial pathology and hip OA: the increase in COL I in OA patients, along with the reduction of COL III and HA, could lead to fascial stiffening, which could alter fascial mechanics and be linked to the development and symptoms of OA. Key Words: Fascia; hip osteoarthritis; hyaluronan; collagen; stiffness; myofascial pain. is the most abundant polysaccharide of the extracellular matrix of connective tissues and the key element that underlies the fascial gliding, lubricating the loose connective tissue between the densely packed collagen layers, between the deep fascia and muscle, and within the muscle itself 1 . It plays also an important role in cell proliferation and mobility, inflammation and angiogenesis, and it is involved in various diseases such as cancer and diabetes 2 . The mean amount of HA in muscular deep fasciae changes according to the various gliding functions to the anatomical site: we found about 43 μg/g of HA in aponeurotic fasciae, which are free to glide over the muscles; there was a drastic decrease (about 6 μg/g) in epimysial fasciae, completely adherent to the underlying muscles, and an increase in the retinacula surrounding joints (90.4 μg/g) 3 . Many physical, mechanical, hormonal and pharmacological factors can influence the production of the various fibrous and glycosaminoglycans components of the fascial extracellular matrix: we found that human fascial fibroblasts are able to produce HA-rich vesicles in vitro within a few hours of cannabinoid CB2 receptor agonist treatment, leading to greater tissue fluidity 4 . In the same way, extracorporeal shock-waves also produce a change in the extracellular matrix and an increase in the HA production, with a reduction in tissue stiffness. Furthermore, viscosity of HA is increased by changes in body temperature (cold) and immobilization. These data can have many clinical implications. For example, an increase in temperature, even of only 2 degrees centigrade, caused by a thermal care, sauna or a deep massage, brings a progressive break-up of the threedimensional superstructure of HA chains, with a consequent decrease in viscosity. At the contrary, cold environment and immobility can increase viscosity of the loose connective tissue, leading to less flexibility of the fasciae and more friction among muscle bundles. Understanding the physicochemical characteristics of HA can help to understand which are the behaviors and the targeted treatments for a healthy fascia. A large number of diseases can affect the peripheral nervous system, with different etiologies and different relative needed managements. The proper diagnosis is necessary for the correct indication of treatment bases on drugs and surgery. Besides these, rehabilitation plays a fundamental role to get the amelioration of functions and quality of life. Among the several approaches, physical agents represent an effective group of conservative methods, which may positively influence nerve recovery. These agents are based on different types of energies (mechanical, electrical, magnetic, thermal) with peculiar indications and targets aimed at pain relief, tissue restoration, neuromuscular stimulation, antiinflammatory action. We performed a multiperspective literature evaluation to present the state of art about the use of physical agents in the rehabilitation of nerve injuries. We collected data about the major studied diseases and the relative effects, the most common outcome measures, the most involved countries in the world about this topic and the modality of the description of the physical agents. We searched in PubMed the terms "physical agents rehabilitation AND peripheral nerv*", finding 131 papers. Our literature review showed the potential benefits of the physical agents even if further studies should be needed for confirmation. Key Words: Rehabilitation; peripheral nerve; physical agents; literature review.

-103 -Extracorporeal Shock Wave Therapy (ESWT) is considered part of the Regenerative Therapy, that represents one of the most challenging branches of rehabilitation field and, more generally, in modern medicine. Basic science studies demonstrated the effectiveness of the ESWT in stimulating biological activities, by the mechanotransduction of the acoustic signal into a biological response, that involves both intra-cell and cell-matrix interactions. According to the Consensus Statement elaborated by ISMST (International Society for Medical Shockwave Treatment) in 2016 and subscribed by SITOD (Italian Society for Shockwave Therapy), some muscular pathologies are among the indications based on clinical experience. Indeed, despite the few studies in the literature, ESWs are widely used in clinical practice in many acquired muscle diseases. The rationale for the use of this treatment is related to its well-known antiinflammatory, analgesic, anti-fibrotic and myorelaxant action. Moreover, ESWT could act on Muscular Satellite Cells, myogenic progenitors, by stimulating them to proliferate, and eventually differentiate through fusion with each other or to damaged fibers to reconstitute fiber integrity and functionality. This hypothesized mechanism may play an important role in helping muscle regeneration, if ESWT is properly inserted in a multimodal rehabilitative approach including therapeutic exercise. However, there is still a lot of confusion about the different types of shock waves (focal, radial), the different generators (electrohydraulic, electromagnetic, piezoelectric), the parameters and treatment protocols. Therefore, this presentation aims to provide an update on the efficacy of extracorporeal shockwave treatment for muscular acquired pathologies by focusing on the evidence.

Key Words: Extracorporeal shock wave therapy (ESWT); muscle; evidence; regenerative medicine.

Can home-based rehabilitation be effective to counteract skeletal muscle atrophy and to ameliorate physical functioning of elderly patients? Elderly subjects with musculoskeletal and neurological diseases spend less time training on a regular basis, worsening their mobility concerns and undergoing muscular atrophy. As a result of the pandemic, access to regular physical activity has reduced even in healthy older individuals, due to mobility restrictions, quarantine, and lockdown. It might be beneficial to provide all these individuals, both affected by chronic disorders and healthy, with rehabilitation program to be admistered at home or even in bed (Full-Body In-Bed Gym). Homebased exercise programs are increasingly being employed in order to improve physical functioning of elderly patients with orthopedic (e.g., hip and knee osteoarthritis) and neurological disorders (e.g., Parkinson's disease, spinal cord injury, etc.). Through a comprehensive review of the literature, we investigated what is the evidence of the efficacy of home-based exercise programs for different subjects and what are the most appropriate modalities to perform these programs. The evidence in literature seems to be promising: homebased treatment seems to be effective, safe and usable by elderly patients suffering from different diseases as well as healthy ones. In the future, home-based exercise programs may represent a valuable option to counteract muscle atrophy and reduced motor function in the elderly, taking also advantage of modern devices such as those employed in telerehabilitation. Purpose of this study was to assess the effect of a course of homeopathic preparations using acupuncture points on the severity of motor impairment against the background of early rehabilitation of the ischaemic stroke (IS). More than 60 patients with different localisation of ischaemic stroke (vertebro-basilar and carotid territories) were treated. All the patients were treated according to the standards of medical care for ischaemic stroke. The degree of impairment was assessed using National Institutes of Health Stroke Scale (NIHSS), which included impairment of consciousness, degree of paresis (0 -no paresis, 4 -no active movements), sensory disturbance and speech disorders manifested in the form of aphasia and dysarthria. The control group consisted of 10 patients with ischaemic stroke. All the patients underwent a computer-assisted tomography of the brain to rule out the hemorrhagic nature of the stroke. Treatment was given during the acute period of IS within 1 to 3 days after onset. Exclusion criteria: hemorrhagic and mixed strokes, thrombolysis, history of cancer. The basic therapy of IS was carried out according to the standards. HEEL homeopathic medicines with antiinflammatory, metabolic and antioxidant effects were used, which were administered on the common action points of the Yang meridians of the paretic limbs. The control group received acupuncture according to the same scheme on the paretic limbs. The treatment duration was 10 sessions; the observation lasted for 21 days, according to the standards of hospitalization for stroke patients. The degree of functional impairment of patients treated with medicinal acupuncture and the patients of the control group assessed by the National Institutes of Health Stroke Scale (NIHSS), (Fig. 1,2) . The outcomes of the medicinal acupuncture treatment: positive effect in the form of a reduction in the degree of paresis (Fig. 3,4) were observed in patients, who received medicinal acupuncture. The lowest treatment outcome was observed in patients with IS in the vertebro-basilar Gym exercises to be practiced in hospital or at home to improve the motor function system. Medicinal acupuncture can be used in early rehabilitation for patients with mild and moderate IS, since its possible mechanism is the enhancement of neuroplasticity in the conditions of brain damage. The use of homeopathic medicines can increase the positive effect compared with acupuncture. The most promising "candidates" for this treatment are the patients with IS in the carotid territory.

Key Words: ischaemic stroke; medicinal acupuncture .

extensive ischemia and micro-thrombosis, myocardial and brain lesions, will require a long period for recovery. 2,3 However, it is possible to optimize this process through sanatorium and spa resort treatment. When the patient arrives at the sanatorium-and-spa institution, his or her condition should be stabilized. The main objectives of spa treatment are to provide general strengthening therapy, symptomatic treatment, prevention of disability and improvement of the quality of life. When organizing sanatorium and spa treatment for patients who have had COVID-19, there are two main components: 1. A spa location characterized by natural healing factors; 2. Areas and methods of sanatorium and spa treatment, in which a nutritional therapy plays an essential role. Particularly, in the case of rehabilitation of patients after a severe and extremely severe course of the disease, who have a weight deficit. In conclusion it should be noted that the sanatorium and spa resort treatment of patients after COVID-19 is defined by the following main areas and methods used for the recovery of this category of patients: 1. Nutritional therapy; 2. Active lifestyle; 3. Natural healing factors; 4. Physiotherapy; 5. Terrainkur; 6. Massage; 7. Aerosol therapy;

8. Photohaemotherapy; 9. Electroneurostimulation of the lymphatic drainage; 10. Lowfrequency magnetic therapy; 11. SMC therapy; 12. Interference therapy; 13. Pulsed low frequency electrostatic field or electrostatic massage; 14. Decimetre-wave band electromagnetic radiation; 15. Vibrotherapy; 16. Laser therapy; 17. Oxygenbarotherapy; 18. Ultrasonic therapy; 19. General magnetotherapy; 20. Carbon dioxide baths; 21. Transcranial electrotherapy; 22. Psychological correction and Psychotherapy; 23. Biofeedback (biofeedback therapy). 4 All of the above-mentioned methods of treatment are effectively used both individually and as part of comprehensive sanatorium and spa resort treatment and medical rehabilitation programmes.

Key Words: Coronavirus infection; rehabilitation treatment; sanatorium and spa resort.

Focal vibration (FV), when applied with specific intensities and frequencies on belly muscle, can improve muscle strengthening inducing peripheral and, in the long term, central modifications. Therefore, this approach is useful when subjects need a strengthening treatment after o during prolonged immobility or disabling diseases that prevent them from performing traditional protocols. Similarly, action observation (AO) and "motor imagery" techniques are used to intensify the effect of rehabilitation on muscle strength and to reduce recovery time. The synergy effect of the two approaches was not demonstrated. The aim of this study is to assess the effectiveness of the integration of a strengthening protocol based on FV associated with an AO paradigm. The study involved healthy subjects, who were randomly assigned to a specific treatment: a group underwent 100 Hz FV treatment on the right femoral quadriceps and a second group underwent the same protocol associated with an AO paradigm of the muscle training (AO+FV) for two weeks, five days/week. Before (T0), after the first week (T1), at the end of treatment (T2), and after a week from the end (T3) for the follow-up, we assessed the maximum voluntary contraction (MVC) expressed at knee extension bilaterally with dynamometer and at T0, T2 and T3 we assessed changes in cortical excitability by transcranial magnetic stimulation (Resting Motor Threshold -RMT; Silent Period -SP). Preliminary data seems to show that MVC increases bilaterally in both groups at T1 and T2. A major increase is evident in the AO+FV group, this increase is maintained even at T3. RTM is reduced at T3 in AO + FV. The SP is reduced only in group AO + FV at T2 and T3. In conclusion, AO seems to be able to enhance the effect of muscle reinforcement induced by FV on maximum strength. This effect seems to be mediated by peripheral and central modifications. More evidence is needed to support treatment effectiveness, which could be a valuable addition to existing protocols as well as an alternative treatment when the subject is not able to perform traditional reinforcement protocols.

Key Words: Parkinson's Disease; postural deviation; Pisa Syndrome; trunk dystonia. successful in reducing the postural abnormality and posture related pain, as suggested by the significative variation of TFA and the major improvement seen in clinical-functional scales .

Key Words: Fascia; hip osteoarthritis; hyaluronan; collagen; stiffness; myofascial pain.

In multiple sclerosis (MS) patients, symptoms such as: fatigue, lack of physical energy, spasticity, difficulty in performing movements, and motor coordination disorders, and with tremors, dizziness and postural instability are among the most common complications. Cattaneo et al. (2007) studied the effects of stability training on MS patients and described it as an effective intervention in reducing falls risk, improving stability, and strength. Therefore, the present study aimed to confirm early observations by the application of a proprioceptive rehabilitation program targeting balance and strength on MS patients. 13 subjects with MS, 5 male and 8 female, volunteered in the study. A MAP hand dynamometer was used to determine the handgrip strength, as the maximum isometric grip (MIG), recognized as an important health indicator for determining musculoskeletal function, as well as weakness and disability. Sensor Medica® systems allowed us to carry out baropodometric and stabilometric measurements using the associated program (Freestep® by Sensor Medica®). The device consists in a pressure platform with resistive sensors with conductive rubber. The rehabilitation protocol included: A) 10 minutes of Motomed; B) 10 minutes of Human Tecar proprioceptive path; C) 15 minutes in total of physical exercises; D) 15 minutes of massage therapy of the whole spine All patients wore the photon emission devices TAOPATCH by TAO technologies, according to a protocol used in a previous study (Amato et al., 2021) . Testing procedures were carried out before and after the rehabilitation protocol. The paired sample t-test revealed statistically significant improvements for the baropodometric measures (p<0.05) associated with the intervention. In particular, plantar surface pressure, forefoot and hindfoot pressure improved significantly, as well as the maximal and mean foot pressures and the forefoot and hindfoot loads ( figure 1a ). In addition, the intervention induced a statistically significant improvement in the right (p = 0.023) and left (p = 0.021) handgrip strength (figure 1b). The combination of an adequate and specific rehabilitation protocol combined with the application of the photon emission devices Taopatch, emerged as a valid protocol to improve handgrip strength of the upper limbs and the expression of the body weight on the ground in contrast with the force of gravity. Thus, determining improvements in a cluster of parameters severely decompensated in MS patientsIn multiple sclerosis (MS) patients, symptoms such as: fatigue, lack of physical energy, spasticity, difficulty in performing movements, and motor coordination disorders, and with tremors, dizziness and postural instability are among the most common complications. Cattaneo et al. (2007) studied the effects of stability training on MS patients and described it as an effective intervention in reducing falls risk, improving stability, and strength. Therefore, the present study aimed to confirm early observations by the application of a In multiple sclerosis (MS) patients, symptoms such as: fatigue, lack of physical energy, spasticity, difficulty in performing movements, and motor coordination disorders, and with tremors, dizziness and postural instability are among the most common complications. Cattaneo et al. (2007) studied the effects of stability training on MS patients and described it as an effective intervention in reducing falls risk, improving stability, and strength. Therefore, the present study aimed to confirm early observations by the application of a proprioceptive rehabilitation program targeting balance and strength on MS patients. 13 subjects with MS, 5 male and 8 female, volunteered in the study. A MAP hand dynamometer was used to determine the handgrip strength, as the maximum isometric grip (MIG), recognized as an important health indicator for determining musculoskeletal function, as well as weakness and disability. Sensor Medica® systems allowed us to carry out baropodometric and stabilometric measurements using the associated program (Freestep® by Sensor Medica®). The device consists in a pressure platform with resistive sensors with conductive rubber. The rehabilitation protocol included: A) 10 minutes of Motomed; B) 10 minutes of Human Tecar proprioceptive path; C) 15 minutes in total of physical exercises; D) 15 minutes of massage therapy of the whole spine All patients wore the photon emission devices TAOPATCH by TAO technologies, according to a protocol used in a previous study (Amato et al., 2021) . Testing procedures were carried out before and after the rehabilitation protocol. The paired sample t-test revealed statistically significant improvements for the baropodometric measures (p<0.05) associated with the intervention. In particular, plantar surface pressure, forefoot and hindfoot pressure improved significantly, as well as the maximal and mean foot pressures and the forefoot and hindfoot loads ( figure 1a ). In addition, the intervention induced a statistically significant improvement in the right (p = 0.023) and left (p = 0.021) handgrip strength (figure 1b). The combination of an adequate and specific rehabilitation protocol combined with the application of the photon emission devices Taopatch, emerged as a valid protocol to improve handgrip strength of the upper limbs and the expression of the body weight on the ground in contrast with the force of gravity. Thus, determining improvements in a cluster of parameters severely decompensated in MS patientsproprioceptive rehabilitation program targeting balance and strength on MS patients. 13 subjects with MS, 5 male and 8 female, volunteered in the study. A MAP hand dynamometer was used to determine the handgrip strength, as the maximum isometric grip (MIG), recognized as an important health indicator for determining musculoskeletal function, as well as weakness and disability. Sensor Medica® systems allowed us to carry out baropodometric and stabilometric measurements using the associated program (Freestep® by Sensor Medica®). The device consists in a pressure platform with resistive sensors with conductive rubber. The rehabilitation protocol included: A) 10 minutes of Motomed; B) 10 minutes of Human Tecar proprioceptive path; C) 15 minutes in total of physical exercises; D) 15 minutes of massage therapy of the whole spine. All patients wore the photon emission devices TAOPATCH by TAO technologies, according to a protocol used in a previous study (Amato et al., 2021) .

Testing procedures were carried out before and after the rehabilitation protocol. The paired sample t-test revealed statistically significant improvements for the baropodometric measures (p<0.05) associated with the intervention. In particular, plantar surface pressure, forefoot and hindfoot pressure improved significantly, as well as the maximal and mean foot pressures and the forefoot and hindfoot loads ( figure 1a ). In addition, the intervention induced a statistically significant improvement in the right (p = 0.023) and left (p = 0.021) handgrip strength (figure 1b). The combination of an adequate and specific rehabilitation protocol combined with the application of the photon emission devices Taopatch, emerged as a valid protocol to improve handgrip strength of the upper limbs and the expression of the body weight on the ground in contrast with the force of gravity. Thus, determining improvements in a cluster of parameters severely decompensated in MS patients.

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The state of helplessness in preschool children with mental retardation and its correction The presentation deals with the problem of helplessness in preschool children with mental retardation (ANW). Became widely known studies of so-called "learned helplessness" Seligmena, in the national psychology studies "of personal helplessness" Ciring, etc., but work in assessing the role and place of helplessness (just helplessness, learned, personal) in preschool children in general and delayed mental development in particular is virtually non-existent. At the base of the research based on the ideas of Vygotsky about the features of development of children, as well as whole-value approach and the biopsychosocionoetic model of human nature, its health and development, proposed by one of the authors of the article. The authors put forward a hypothesis to test done quite voluminous study of the phenomenon of helplessness, which brought together 140 children and their families (mothers). Of these, 90 children with CRA and 50 without such a diagnosis. 45 children with CRA entered the pilot group, which on the second stage of the study, participated in the psychocorrective program proposed by the authors of the study. The monitoring group comprised 45 children with CRA and 50 children without the CRA. Based on the quantitative and qualitative analyses of the results obtained was differentially 3 groups of children with varying degrees of severity, the condition of helplessness: no signs of the condition of helplessness, with moderately expressed signs of helplessness and the lack thereof. The results of the second cut after the implementation of the program of psychocorrective indicate the possibility, under certain conditions, absorb any movement state of helplessness.

Key Words: Helplessness; learned helplessness; personal helplessness; children of pre-school age; development; techniques; attitude; family; parents; psychological support.

2021 semi-virtual PDM3 from Thermae of Euganean Hills

30 Years of Translational Mobility Medicine: 2020 Padua Muscle Days go virtual from Euganean Hills

30-years of Translational Mobility Medicine at the time of COVID-19 outbreak: Last-minute forewords from the editor

Thirty years of translational research in Mobility Medicine: Collection of Abstracts of the 2020 Padua Muscle Days

Cancer-associated cachexia. Nat Rev Dis Primers

Definition and classification of cancer cachexia: an international consensus

Skeletal Muscle Pathology in Peripheral References 1. Vassilakos G, Barton ER. Insulin-Like Growth Factor I Regulation and Its Actions in Skeletal Muscle

The pro-forms of insulin-like growth factor I (IGF-I) are predominant in skeletal muscle and alter IGF-I receptor activation

E-peptides control bioavailability of IGF-1

Mutational analysis of the insulin-like growth factor I prohormone processing site

Ischaemic accumulation of succinate controls reperfusion injury through mitochondrial ROS

High-fat diet reduces the formation of butyrate, but increases succinate, inflammation, liver fat and cholesterol in rats, while dietary fibre counteracts these effects

Influence of diabetes mellitus duration on the efficacy of ischemic preconditioning in a Zucker diabetic fatty rat model

pH-Gated Succinate Secretion Regulates Muscle Remodeling in Response to Exercise

The wasting-associated metabolite succinate disrupts myogenesis and impairs skeletal muscle regeneration

Skeletal myopathy in a rat model of postmenopausal heart failure with preserved ejection fraction

PDM3 On-site •

Mitochondrial DNA damage in calf skeletal muscle and walking performance in people with peripheral artery disease. Free Radic Biol Med

Associations of Peripheral Artery Disease With Calf Skeletal Muscle Mitochondrial DNA Heteroplasmy

Associations of Poly (ADP-Ribose) Polymerase1 abundance in calf skeletal muscle with walking performance in peripheral artery disease

The p97/VCP ATPase is critical in muscle atrophy and the accelerated degradation of muscle proteins

Amyotrophic Lateral Sclerosis: An Update for 2018

Alcohol-abuse drug disulfiram targets cancer via p97 segregase adaptor NPL4

PDM3 On-site •

Definition and classification of cancer cachexia: an international consensus

Cancer-associated cachexia

Cachexia as a major underestimated and unmet medical need: facts and numbers

Cancer cachexia and fat-muscle physiology

Comparing two

How much dystrophin is enough: the physiological consequences of different levels of dystrophin in the mdx mouse

Changes in Myonuclear Number During Postnatal Growth -Implications for AAV Gene Therapy for Muscular Dystrophy

Proteomic analysis identifies key differences in the cardiac interactomes of dystrophin and micro-dystrophin

AAV Vector Immunogenicity in Humans: A Long Journey to Successful Gene Transfer

PDM3 On-site •

Absence of Significant Off-Target Splicing Variation with a U7snRNA Vector Targeting DMD Exon 2 Duplications. Hum Gene Ther

Pre-clinical dose-escalation studies establish a therapeutic range for U7snRNA-mediated DMD exon 2 skipping

Lack of Toxicity in Nonhuman Primates Receiving Clinically Relevant Doses of an AAV9.U7snRNA Vector Designed to Induce DMD Exon 2 Skipping. Hum Gene Ther

Italian DMD Network and the United Dystrophinopathy Project. Phenotypic Spectrum of Dystrophinopathy Due to Duchenne Muscular Dystrophy Exon 2 Duplications

Dystrophin: the protein product of the Duchenne muscular dystrophy locus

Dystrophin protects the sarcolemma from stresses developed during muscle contraction

Association of dystrophin-related protein with dystrophin-associated proteins in mdx mouse muscle

A-and B-utrophin have different expression patterns and are differentially up-regulated in mdx muscle

Adeno-associated virus vector carrying human minidystrophin genes effectively ameliorates muscular dystrophy in mdx mouse model

Modular flexibility of dystrophin: implications for gene therapy of Duchenne muscular dystrophy

Skeletal and cardiac

Oculopharyngeal Muscular Dystrophy (OPMD)

Involvement of the ubiquitinproteasome pathway and molecular chaperones in oculopharyngeal muscular dystrophy

Deregulation of the ubiquitin-proteasome system is the predominant molecular pathology in OPMD animal models and patients. Skelet Muscle

Dysfunctional transcripts are formed by alternative polyadenylation in OPMD

Pharmacological modulation of the ER stress response ameliorates oculopharyngeal muscular dystrophy

2022 PDM3 On-site •

Improving clinical trial outcomes in amyotrophic lateral sclerosis

Skeletal Muscle in ALS: An Unappreciated Therapeutic Opportunity? Cells

Amyotrophic lateral sclerosis is a distal axonopathy: evidence in mice and man

2022 PDM3 On-site •

Dieter Swandulla 3, Kay Ohlendieck 1,2 References 1. Ohlendieck K, Swandulla D. Complexity of skeletal muscle degeneration: multi-systems pathophysiology and organ crosstalk in dystrophinopathy

Mass Spectrometric Profiling of Extraocular Muscle and Proteomic Adaptations in the mdx-4cv Model of Duchenne Muscular Dystrophy

Proteomic profiling of the interface between the stomach wall and the pancreas in dystrophinopathy

Proteome-wide Changes in the mdx-4cv Spleen due to Pathophysiological Cross Talk with Dystrophin-Deficient Skeletal Muscle

Proteomic profiling of the mouse diaphragm and refined mass spectrometric analysis of the dystrophic phenotype

Duchenne Muscular Dystrophy

Very mild muscular dystrophy associated with the deletion of 46% of dystrophin

Muscular dystrophies. Lancet

Gene Therapy for Duchenne Muscular Dystrophy

Expanding AAV packaging capacity with trans-splicing or overlapping vectors: a quantitative comparison

2022 PDM3 On-site •

Developmental and regenerative paradigms of cilia regulated hedgehog signaling

Ciliary Hedgehog Signaling Restricts Injury-Induced Adipogenesis

Sonic hedgehog therapy in a mouse model of age-associated impairment of skeletal muscle regeneration

Hedgehogdependent regulation of angiogenesis and myogenesis is impaired in aged mice

2022 PDM3 On-site •

Neutral Lipid Storage Diseases as Cellular Model to Study Lipid Droplet Function

MiRNAs as biomarkers of phenotype in neutral lipid storage disease with myopathy

Neutral lipid storage disease with subclinical myopathy due to a retrotransposal insertion in the PNPLA2 gene

Metabolic lipid muscle disorders: biomarkers and treatment

PDM3 On-site •

Inter-tissue communication in cancer cachexia

Cancer-associated cachexia. Nat Rev Dis Primers

Mop3 is an essential component of the master circadian pacemaker in mammals

Early aging and agerelated pathologies in mice deficient in BMAL1, the core componentof the circadian clock

Dissecting the functions of the mammalian clock protein BMAL1 by tissue-specific rescue in mice

Defining the Independence of the Liver Circadian Clock

Intrinsic muscle clock is necessary for musculoskeletal health

PDM3 On-site •

Deletion of muscle IGF-I transiently impairs growth and progressively disrupts glucose homeostasis in male mice

Localized Igf-1 transgene expression sustains hypertrophy and regeneration in senescent skeletal muscle

Insulin inhibits AMPK activity and phosphorylates AMPK Ser⁴⁸⁵/⁴⁹¹ through Akt in hepatocytes, myotubes and incubated rat skeletal muscle

2022 PDM3 On-site •

Transcriptomic profiling of skeletal muscle adaptations to exercise and inactivity

Time trajectories in the transcriptomic response to exercise -a meta-analysis

SpillOver stimulation: A novel hypertrophy model using co-contraction of the plantar-flexors to load the tibial anterior muscle in rats

Master Regulator of the Cancer Epigenome and Transcriptome. Genes (Basel)

2022 PDM3 On-site •

Peroxisome biogenesis disorders. Transl Sci Rare Dis

Structural biology of the import pathways of peroxisomal matrix proteins

Skeletal Muscle Regulates Metabolism via Interorgan Crosstalk: Roles in Health and Disease

PDM3 On-site •

Consensus definition of sarcopenia, cachexia and precachexia: joint document elaborated by Special Interest Groups (SIG) "cachexia-anorexia in chronic wasting diseases" and "nutrition in geriatrics

Fig 1. Perturbation of BMP signaling pathway leads to muscle wasting in cancer cachexia

Definition and classification of cancer cachexia: an international consensus

Cancer-associated cachexia. Nat Rev Dis Primers

Cachexia as a major underestimated and unmet medical need: facts and numbers

Cancer cachexia and fat-muscle physiology

Comparing two classifications of cancer cachexia and their association with survival in patients with unresected pancreatic cancer

BMP signaling controls muscle mass

The bone morphogenetic protein axis is a positive regulator of skeletal muscle mass

Perturbed BMP signaling and denervation promote muscle wasting in cancer cachexia

PDM3 On-site •

Deletion of muscle IGF-I transiently impairs growth and progressively disrupts glucose homeostasis in male mice

Actions and interactions of IGF-I and MMPs during muscle regeneration

Matrix Metalloproteinase 13 from Satellite Cells is Required for Efficient Muscle Growth and Regeneration

Gamma-sarcoglycan is required for the response of archvillin to mechanical stimulation in skeletal muscle

PDM3 On-site •

Skeletal muscle V o2 kinetics by the NIRS repeated occlusions method during the recovery from cycle ergometer exercise

Peripheral impairments of oxidative metabolism after a 10-day bed rest are upstream of mitochondrial respiration

PDM3 On-site •

Neuromuscular junction instability and altered intracellular calcium handling as early determinants of force loss during unloading in humans

Skeletal muscle adaptations to physical inactivity and subsequent retraining in young men

Effects of immobilization on contractile properties, recruitment and firing rates of human motor units

2022 PDM3 On-site •

α-motor are spared from aging while their synaptic inputs degenerate in monkeys and mice

Pharmacologic treatment with OKN-007 reduces alpha-motor neuron loss in spinal cord of aging mice

Molecular changes associated with spinal cord aging

Sarcopenia: Neurological Point of View

Perisynaptic Schwann Cells at the Neuromuscular Synapse: Adaptable, Multitasking Glial Cells

Attenuation of agerelated changes in mouse neuromuscular synapses by caloric restriction and exercise

Opposite Synaptic Alterations at the Neuromuscular Junction in an ALS Mouse Model: When Motor Units Matter

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

PDM3 On-site •

Smoke-induced neuromuscular junction degeneration precedes the fibre type shift and atrophy in chronic obstructive pulmonary disease

Fidelity of muscle fibre reinnervation modulates ageing muscle impact in elderly women

Is age-related decline in lean mass and physical function accelerated by obstructive lung disease or smoking? Thorax

Kynurenines link chronic inflammation to functional decline and physical frailty

Chronic aryl hydrocarbon receptor activity phenocopies smoking-induced skeletal muscle impairment. J Cachexia Sarcopenia Muscle

PDM3 On-site •

Exercisemediated reinnervation of skeletal muscle in elderly people: An update

Reinnervation of Vastus lateralis is increased significantly in seniors (70-years old) with a lifelong history of high-level exercise

Use it or Lose It: Tonic Activity of Slow Motoneurons Promotes Their Survival and Preferentially Increases Slow Fiber-Type Groupings in Muscles of Old Lifelong Recreational Sportsmen

Long-term high-level exercise promotes muscle reinnervation with age

Recovery from muscle weakness by exercise and FES: lessons from Masters, active or sedentary seniors and SCI patients

PDM3 On-site •

Cardiovascular and skeletal muscle health with lifelong exercise

What is the effect of ageing on type 2 muscle fibres?

What is the cause of the ageing atrophy? Total number, size and proportion of different fiber types studied in whole vastus lateralis muscle from 15-to 83-year-old men

Specific contributions of various muscle fibre types to human muscle performance: an in vitro study

Skeletal muscle signature of a champion sprint runner

Single muscle fibre contractile characteristics with lifelong endurance exercise

MRC-Versus Arthritis Centre of Excellence for Musculoskeletal Ageing Research

Running performance differences between men and women:an update

Do older athletes reach limits in their performance during marathon running? Age (Dordr)

Rate and mechanism of maximal oxygen consumption decline with aging: implications for exercise training

Cardiovascular and skeletal muscle health with lifelong exercise

Age-associated power decline from running, jumping, and throwing male Masters World Records

Recovery from muscle weakness by exercise and FES: lessons from Masters, active or sedentary seniors and SCI patients

Master World Records show minor gender differences of performance decline with aging

In vivo intramuscular fascicle-aponeuroses dynamics of the human medial gastrocnemius during plantarflexion and dorsiflexion of the foot

Asymmetric deformation of contracting human gastrocnemius muscle

Age-related differences in strain rate tensor of the medial gastrocnemius muscle during passive plantarflexion and active isometric contraction using velocity encoded MR imaging: potential index of lateral force transmission

PDM3 On-site •

Age-related differences in strain rate tensor of the medial gastrocnemius muscle during passive plantarflexion and active isometric contraction using velocity encoded MR imaging: potential index of lateral force transmission

Relationship of changes in strain rate indices estimated from velocity-encoded MR imaging to loss of muscle force following disuse atrophy

Compressed sensing velocity encoded phase contrast imaging: Monitoring skeletal muscle kinematics

Aortic 4D flow MRI in 2 minutes using compressed sensing, respiratory controlled adaptive k-space reordering, and inline reconstruction

PDM3 On-site •

Exercise-dependent formation of new junctions that promote STIM1-Orai1 assembly in skeletal muscle. Sci Rep

STIM1, an essential and conserved component of store-operated Ca2+ channel function

A mutation in Orai1 causes immune deficiency by abrogating CRAC channel function

Depletion of Ca 2+ in the sarcoplasmic reticulum stimulates Ca 2+ entry into mouse skeletal muscle fibres

Pre-assembled Ca2+ entry units and constitutively active Ca2+ entry in skeletal muscle of calsequestrin-1 knockout mice

Transverse tubule remodeling enhances Orai1-dependent Ca 2+ entry in skeletal muscle. Elife

PDM3 On-site •

How Long Does Muscle Memory Last? The Role of Epigenetics

Specific labelling of myonuclei by an antibody against pericentriolar material 1 on skeletal muscle tissue sections

Lamin-Related Congenital Muscular Dystrophy Alters Mechanical Signaling and Skeletal Muscle Growth

SpillOver stimulation: A novel hypertrophy model using co-contraction of the plantar-flexors to load the tibial anterior muscle in rats

The breaking and making of healthy adult human skeletal muscle in vivo. Skelet Muscle

PDM3 On-site •

Merg1a K+ channel induces skeletal muscle atrophy by activating the ubiquitin proteasome pathway

MERG1A Protein Abundance Increases in the Atrophied Skeletal Muscle of Denervated Mice, But Does Not Affect NFκB Activity

The ERG1a potassium channel increases basal intracellular calcium concentration and calpain activity in skeletal muscle cells. Skelet Muscle

Investigation of the mechanism by which the ERG1a K + channel increases intracellular calcium. Presented at 2018 Spring Padua Muscle Days

A 3D diffusional-compartmental model of the calcium dynamics in cytosol, sarcoplasmic reticulum and mitochondria of murine skeletal muscle fibers

Simultaneous recording of calcium transients in skeletal muscle using high-and low-affinity calcium indicators

Parvalbumin affects skeletal muscle trophism through modulation of mitochondrial calcium uptake

Ryanodine receptor leak triggers fiber Ca2+ redistribution to preserve force and elevate basal metabolism in skeletal muscle. Sci Adv

PDM3 On-site •

Aetiology and natural history of Parkinson's disease

A gait analysis data collection and reduction technique

Gait analysis may distinguish progressive supranuclear palsy and Parkinson disease since the earliest stages. Sci Rep

PDM3 On-site •

A Novel Knee Bone and Cartilage Osteoarthritis Index Extracted from a Patient-Specific Image Feature Analysis

Epidemiology of osteoarthritis

Radiological assessment of osteo-arthrosis

Osteoarthrosis of the knee. A radiographic investigation

PDM3 On-site •

Postural Control Assessment through Visual Induced Motion Sickness and a Moving Force Platform Andrea Colacino 1,2, Romain Aubonnet 1

Toward Predicting Motion Sickness Using Virtual Reality and a Moving Platform Assessing Brain, Muscles, and Heart Signals

Postural Control Adaptation and Habituation During Vibratory Proprioceptive Stimulation: An HD-EEG Investigation of Cortical Recruitment and Kinematics

PDM3 On-site •

CT-and MRI-Based 3D Reconstruction of Knee Joint to Assess Cartilage and Bone. Diagnostics (Basel)

A CTbased method to compute femur remodelling after total hip arthroplasty

PDM3 On-site •

Global, regional, and national incidence, prevalence, and years lived with disability for 301 acute and chronic diseases and injuries in 188 countries, 1990-2013: a systematic analysis for the Global Burden of Disease Study

The Fly": a new clinical assessment and treatment method for deficits of movement control in the cervical spine: reliability and validity. Spine (Phila Pa 1976)

Comparison of patient-and clinician-reported outcome measures in lower back rehabilitation: introducing a new integrated performance measure (t2D)

Machine learning approaches to predict rehabilitation success based on clinical and patient-reported outcome measures

General and Disease-Specific Health Indicator Changes Associated with Inpatient Rehabilitation

Dynamics of Vagal Activity Due to Surgery and Subsequent Rehabilitation. Front Neurosci

Rehabilitation and Health": URL: www.researchgate.net/project/Rehabilitation-and-Health

PDM3 On-site •

Effectiveness of eccentric-biased exercise interventions in reducing the incidence of falls and improving functional performance in older adults: a systematic review

Effects of Eccentric-Focused Versus Conventional Training on Lower Limb Muscular Strength in Older People: A Systematic Review With Meta-Analysis

PDM3 On-site •

Mimicking disuse and rehabiliation in a mouse model Feliciano Protasi 2022 PDM3 On-site •

Email: sanzamp@unipd.it 2022 PDM3 On-site •

Avoided heat-related mortality through climate adaptation strategies in three US cities

Changes in hot days and heat waves in China during 1961-2007

Increasing trends in regional heatwaves

Cities must protect people from extreme heat

Homogenization of instrumental time series of air temperature in Central Italy

PDM3 On-site •

Environmental and Exertional Heat Stroke: the role of skeletal muscle Feliciano Protasi CAST

Death toll exceeded 70,000 in Europe during the summer of Fig 1. Environmental and Exertional Heat Stroke: Rational of the role of skeletal muscle

Heat stroke

Heat stroke: a comprehensive review

RyR1 S-nitrosylation underlies environmental heat stroke and sudden death in Y522S RyR1 knockin mice

The importance of calcium ions for in vitro malignant hyperthermia testing

High-fat diet increases the risk of environmental heatstroke in mice

Email: matteo.serano@unich.it References 1. Bouchama A, Knochel JP. Heat stroke

RyR1 S-nitrosylation underlies environmental heat stroke and sudden death in Y522S RyR1 knockin mice

Increased oxidative stress precedes the onset of high-fat dietinduced insulin resistance and obesity

2022 PDM3 On-site •

Heat stroke

RyR1 S-nitrosylation underlies environmental heat stroke and sudden death in Y522S RyR1 knockin mice

Exercise-dependent formation of new junctions that promote STIM1-Orai1 assembly in skeletal muscle. Sci Rep

Calcium entry units (CEUs): perspectives in skeletal muscle function and disease

Age-related changes in the rate of muscle activation and rapid force characteristics

The aging neuromuscular system and motor performance

Rate of force development: physiological and methodological considerations

Rate of Force Development as an Indicator of Neuromuscular Fatigue: A Scoping Review. Front Hum Neurosci

2022 PDM3 On-site •

1-year outcomes in hospital survivors with COVID-19: a longitudinal cohort study

Solve ME. Long Covid Alliance

Adaptive Force and emotionally related imaginations -preliminary results suggest a reduction of the maximal holding capacity as reaction to disgusting food imagination. Heliyon

Disgusting odours affect the characteristics of the Adaptive Force in contrast to neutral and pleasant odours. Sci Rep

PDM3 On-site •

Bipolar transcutaneous spinal stimulation evokes short-latency reflex responses in human lower limbs alike standard unipolar electrode configuration

Comparison of Twitch Responses During Currentor Voltage-Controlled Transcutaneous Neuromuscular Electrical Stimulation

Electrical Nerve Stimulation: Theory, Experiments, and Applications

Neurophysiology of epidurally evoked spinal cord reflexes in clinically motor-complete posttraumatic spinal cord injury

Structural differentiation of skeletal muscle fibers in the absence of innervation in humans

Recovery of long-term denervated human muscles induced by electrical stimulation. Muscle Nerve

Persistent Muscle Fiber Regeneration in Long Term Denervation. Past, Present, Future

Electrical stimulation of denervated muscles: first results of a clinical study

Home-Based Functional Electrical Stimulation of Human Permanent Denervated Muscles: A Narrative Review on Diagnostics, Managements, Results and Byproducts Revisited 2020. Diagnostics (Basel)

2022 PDM3 On-site •

Making muscle: skeletal myogenesis in vivo and in vitro

Towards stem cell therapies for skeletal muscle repair

Satellite cell characterization from aging human muscle

Current Methods for Skeletal Muscle Tissue Repair and Regeneration

Future research and therapeutic applications of human stem cells: general, regulatory, and bioethical aspects

Nanodecoration of electrospun polymeric fibers with nanostructured silver coatings by ionized jet deposition for antibacterial tissues

2022 PDM3 On-site •

Cell-microenvironment interactions and architectures in microvascular systems

Fibrin-based delivery strategies for acute and chronic wound healing

Hydrostatic pressure-generated reactive oxygen species induce osteoarthritic conditions in cartilage pellet cultures

PDM3 On-site •

References 1. Kovalchuk VV. Basic Theoretical and Practical Aspects of Neuro-rehabilitation

Relationship between Movement Stereotype and Focus Location in the Early Recovery Period after Mild Ischemic Stroke

The Importance of Physical Activity in the Rehabilitation of Patients with Cardiovascular Diseases

PDM3 On-site •

Head and neck functional analysis: the Functional Anatomy Research Center experience

Immediate effect of a stabilization splint on masticatory muscle activity in temporomandibular disorder patients

Fatigue in the sternocleidomastoid muscle and hip dysplasia: a surface electromyographic assessment in adult women

Electromyographic assessment of jaw muscles in patients with All-on-Four fixed implantsupported prostheses

Abnormal variations in the facial soft tissues of individuals with Down Syndrome: Sudan versus Italy

Preliminary approach for the surface electromyographical evaluation of the oral phase of swallowing

The efficacy of myofunctional therapy in patients with atypical swallowing

PDM3 On-site •

Equilibrium theory revisited: factors influencing position of the teeth

The role of soft tissues in the aetiology of malocclusion. Dent Update

Follow-up study of functional and morphological malocclusion trait changes from 3 to 12 years of age

The efficacy of myofunctional therapy in patients with atypical swallowing

Orthodontic treatment for prominent upper front teeth (Class II malocclusion) in children and adolescents. Cochrane Database Syst Rev

Contemporary orthodontics

2022 PDM3 On-site •

Evaluation of minimum interdental threshold ability in dentate female temporomandibular disorder patients

SEMG activity of jaw-closing muscles during biting with different unilateral occlusal supports

Occlusal features and masticatory muscles activity. A review of electromyographic studies

Evidence of an influence of asymmetrical occlusal interferences on the activity of the sternocleidomastoid muscle

The dental occlusion as a suspected cause for TMDs: epidemiological and etiological considerations

Effect of artificial occlusal interferences depends on previous experience of temporomandibular disorders

Biomechanical model of the human mandible in unilateral clench: distribution of temporomandibular joint reaction forces between working and balancing sides

PDM3 On-site •

Clinical guidelines for the prevention and treatment of osteoporosis: summary statements and recommendations from the Italian Society for Orthopaedics and Traumatology

The burden of musculoskeletal conditions at the start of the new millennium

Contributions to lateral balance control in ambulatory older adults

Physical Exercise in Aging: Nine Weeks of Leg Press or Electrical Stimulation Training in 70 Years Old Sedentary Elderly People

PDM3 On-site •

Osteoarthritis and inflammation: A serious disease with overlapping -101 -phenotypic patterns

Systematic Review of the Prevalence of Radiographic Primary Hip Osteoarthritis

Study of Osteoporotic Fractures Research Group. Hip Osteoarthritis and the Risk of All-Cause and Disease-Specific Mortality in Older Women: A Population-Based Cohort Study

Prevalence of knee and hip osteoarthritis and the appropriateness of joint replacement in an older population

PDM3 On-site •

Caterina Fede 1, Ilaria Fortunati 2, Carmelo Pirri 1

Hyaluronan and the Fascial Frontier

A Closer Look at the Cellular and Molecular Components of the Deep/Muscular Fasciae

Quantification of hyaluronan in human fasciae: variations with function and anatomical site

Sensitivity of the Fasciae to the Endocannabinoid System: Production of Hyaluronan-Rich Vesicles and Potential Peripheral Effects of Cannabinoids in Fascial Tissue

2022 PDM3 On-site •

Neurological Manifestations in COVID-19 Patients: A Meta-Analysis

Non-invasive neuromodulation effects on painful diabetic peripheral neuropathy: a systematic review and meta-analysis. Sci Rep

Peripheral Neuropathies Seen by Ultrasound: A Literature Analysis through Lexical Evaluation, Geographical Assessment and Graph Theory. Brain Sci

PDM3 On-site •

The effect of low intensity shockwave treatment (Li-SWT) on human myoblasts and mouse skeletal muscle

Extracorporeal shock wave therapy in musculoskeletal disorders: a review

Satellite Cells and Skeletal Muscle Regeneration

PDM3 On-site •

Age-associated power decline from running, jumping, and throwing male masters world records

Skeletal muscle weakness in older adults homerestricted due to COVID-19 pandemic: a role for fullbody in-bed gym and functional electrical stimulation

Skeletal muscle weakness in older adults homerestricted due to COVID-19 pandemic: a role for fullbody in-bed gym and functional electrical stimulation

To Contrast and Reverse Skeletal Muscle Atrophy by Full-Body In-Bed Gym, a Mandatory Lifestyle for Older Olds and Borderline Mobility-Impaired Persons

Early rehabilitation of ischaemic stroke with medicinal acupuncture: A clinical study

Oleynikova 1, Oganes L. Badalyan 1, Georgiy G. Avakyan 1, Kirill V. Terentev 2

Reflexotherapy for Diseases of the Nervous System

Traditional and Modern Aspects of Oriental Reflexotherapy

Part 2 Basic Instruction. Department of Health and Human Services, National Institute of Neurological Disorders and Stroke. The National Institute of Neurological Disorders and Stroke (NINDS) Version 2

Effect of Acupuncture on Neuroplasticity of Stroke Patients with Motor Dysfunction: A Meta-Analysis of fMRI Studies Neural Plast

Can aqua exercises in fresh water improve the gait stereotype function in patients with a neurological disease?

Ivanova National medical research center of rehabilitation and balneology

Vestibular and corticospinal control of human body orientation in the gravitational field

Arterial hypertension in adults

Headache and arterial hypertension

The Accuracy of the Microsoft Kinect V2 Sensor for Human Gait Analysis. A Different Approach for Comparison with the Ground Truth

Temporary guidelines "Prevention, diagnosis and treatment of new coronavirus infection (COVID-19)

Rehabilitation of COVID-19 patients

The role of physical and rehabilitation medicine in the COVID-19 pandemic: the clinician's view

ZhonghuaJie He He Hu Xi Za Zhi Recommendations for respiratory rehabilitation of coronavirus disease 2019 in adult

Focal Muscle Vibration and Action Observation: a novel approach for muscle strengthening

Physical Medicine and Rehabilitation School, Department of Translational Research and New Technologies in Medicine and Surgery

Efficacy of a combined therapeutic approach in the management of Pisa Syndrome

Pisa Syndrome in Parkinson's Disease: Electromyographic Aspects and Implications for Rehabilitation

Long-term effect of repeated lidocaine injections into the external oblique for upper camptocormia in Parkinson's disease

Martina Cafalli 1, Elisa Doro 1, Fulvio Vercillo 1, Stefania Dalise 2, Carmelo Chisari 1

Falls Efficacy Scale (FES-I). Numerical Rating Scale (NRS); -Truncal flexion angle (TFA) measurement with "Image J" system. One week after T0, patients underwent a threeweek specific high intensity rehabilitation program (trunk muscles stretching, balance and postural control) structured in 5 session per week. At the end of treatment (T1) initial evaluations were repeated. Nine patients were enrolled and completed the protocol (age 71± 4,1 y; M: F=5:4). The comparison of the T0 and T1 data displayed a significant reduction of TFA

In conclusion, the results seem to confirm the importance of EOM in PS. Our protocol was well tolerated and showed to be Fig 1. The comparison of the T0 and T1 data displays a significant reduction of Truncal flexion angle (TFA)

Osteoarthritis and inflammation: A serious disease with overlapping phenotypic patterns

Systematic Review of the Prevalence of Radiographic Primary Hip Osteoarthritis

Study of Osteoporotic Fractures Research Group. Hip Osteoarthritis and the Risk of All-Cause and Disease-Specific Mortality in Older Women: A Population-Based Cohort Study

Prevalence of knee and hip osteoarthritis and the appropriateness of joint replacement in an older population

Diego Genua 1,2, Vincenzo Francavilla 4, Angelo Iovane 1, Giuseppe Messina 1,2 1 Department of Psychological, Pedagogical and Educational Sciences

Effects of balance exercises on people with multiple sclerosis: a pilot study

Taopatch® combined with home-based training protocol to prevent sedentary lifestyle and biochemical changes in MS patients during COVID-19 pandemic

Physical activity and multiple sclerosis: a meta-analysis

Psychiatric disorders and stress factors experienced by staff members in cancer hospitals: a preliminary finding from psychiatric consultation service at National Cancer Center Hospitals in Japan

Evaluating the cross-cultural validity of the Polish version of the Four-Dimensional Symptom Questionnaire (4DSQ) using differential item functioning (DIF) analysis. Fam Pract

Theoretical and methodological contradictions of the formation of the biopsychosocial paradigm

Assessing psychological health in midwifery practice: a validation study of the Four-Dimensional Symptom Questionnaire (4DSQ), a Dutch primary care instrument. Midwifery

Disutility of illness for caregivers and families: a systematic review of the literature

The concept of the reflex in the description of behavior

Effectiveness of a psychoeducation program on learned helplessness and irrational beliefs. Educational Sciences: Theory and Practice

Some exercises with the learned helpessness scale (lh/scale) in hindi

Learned helplessness: unique features and translational value of a cognitive depression model

Attributional style for negative events: A proposition for a more valid and reliable measaure of attributional style

Accepted for publication

The authors thank Attendees, Chairs and Organizers, who succeed in dealing with problems of the COVID pandemics. Acknowledgements: Funding by ASI, NUTRISS and 

*****

The effect of direct muscle stimulation on denervated gluteal muscles and tissue composition in people with chronic spinal cord injury -preliminary results

International FES Centre ® -Swiss Paraplegic Centre, Nottwil, Switzerland Ines Bersch, Email: ines.bersch@paraplegie.ch Electrical stimulation of denervated muscles with long pulses to convert connective and adipose tissue into contractile muscle tissue has been investigated in persons with chronic spinal cord injury. 1 niche-like environment combining biomaterials (tissue engineering) and cells (cell-based therapy). Tissue engineering (TE), in particular, has been recognized as a promising tool and a new therapeutic approach for the renewal of SKM tissue. Biomaterials for SKM TE designed in this work are able to persist long enough to support functional SKM tissue formation, while degrading gradually. The device developed is a biocompatible scaffold composed of aligned nanofibers obtained through the electrospinning technique. The alignment provides the anisotropy of SKM tissue mimicking its natural morphology. Moreover, the scaffold acts as carrier for cell vitality, proliferation and myogenic differentiation. Nanofibrous scaffold endows SKM tissue morphogenesis into parallel-oriented myotubes, promoting early cell cycle exit and contributing to an early myoblast differentiation and fusion into myotubes in comparison to conventional monolayer myoblast culture. The physiological functions of the human body represent the expression of biological phenomena easily observable, but hardly quantifiable in their entirety as they result from the dynamic interaction of multiple factors. Each phenomenon may present in different ways through a different combination of the same parameters that characterize it at different times. Therefore, the evaluation of a biological phenomenon implies the An open descriptive study was carried out, which involved 12 patients (7 men, 5 women) aged between 40 and 62 years, with disorders of maintaining the vertical posture, which correspond to the functional diagnosis coded according to ICF "Gait Stereotype Function" b770.1 -minor disorders (5-24%). The test group (n = 12), in addition to basic therapy and exercises with a physiotherapy instructor, did aqua exercises in fresh water for two weeks (30 minutes, 6 days a week). Evaluation technique: the study before and after the rehabilitation course was carried out using "Habilect" video analysis system. To assess the significance of differences between the groups of qualitative variables χ2 test was used. To assess the significance of differences of quantitative variables of the two test groups Wilcoxon rank sum test was used. The significance of differences is considered to be established at p<0, 05. The study calculated the amplitudes of deviations of the body and the head along Х, Y, Z axes before and after the exercises, as well as deviations of the body motion vector before and after the exercises. While calculating the increase of deviation (deviations of the main body axes from the reference value) using Wilcoxon rank sum test, statistically significant deviations along the X-axis were found (an increase by 306,5%, р=0,0504), along the Zaxis (an increase by 112,68%, р=0,0225) and the Body Angle parameter (an increase by 1973,86% р=0,0323). While calculating the increase of deviations of the head axis from the reference value using Wilcoxon rank sum test, statistically significant deviations along the X-axis were found (an increase by 163,04%, р=0,0280), along the Y-axis (an increase by 85,71%, р=0,0199) and Zparameter (an increase by 173,34% р=0,0292) Thus, aqua exercises, due to a decrease of the deviations amplitude along the three axes (Z, Y, X), contribute to the adjustment of disorders of maintaining the vertical posture, a statistically significant decrease in the amplitude of head and neck movements. 1 There is no reason to doubt that for COVID-19 survivors, too, the possibilities of recovery in sanatorium and spa resort environment will be successfully realized. Although the effects of the SARS-CoV-2 virus on the human organism are, in a sense, unique, the pathological processes that occur undergo the "traditional" stages of regression. The procedures offered by sanatoriums for the treatment of diseases of the respiratory, nervous and cardiovascular systems usually have broad indications and can be offered for recovery to patients who have had COVID-19. The provision of spa treatment and rehabilitation activities is crucial for the effective completion of the recovery process and rehabilitation after COVID-19. Rehabilitation in a sanatorium and spa resort environment enables a comprehensive and effective recovery process for the patient, and for the healthcare system it is an opportunity to relieve the burden on specialised in-patient rehabilitation facilities and polyclinic structures. A study of pathogenesis of pathological processes in COVID-19, morpho-functional and metabolic changes shows that the regression of multi-organ lesions caused by SARS-CoV-2: fibrosis and granulation changes in lungs, respiratory failure, The aim of the study is to study the features of the mental health of the closest relatives of oncohematological patients. The sample consisted of 274 subjects aged 18 to 65 years. The experimental groups were patients and their closest relatives at different periods of the course of the disease: at the onset, at three and six months. The number of people in the sample under consideration progressively decreased due to the mortality rate of patients with the underlying disease (92 at the onset of the disease to 70 people on each side at the end of the study). The following research methods were used: clinical and psychological (a conversation aimed at collecting psychological anamnesis and identifying biosocial characteristics of patients, assessment of the current mental status, assessment of the severity of the condition-the Karnovsky index); testing (methods SF-36, MFA-20, 4DSQ, coping behavior in stressful situations by Norman S.); instrumental studies (assessment of the Kerdo vegetative index); mathematical and statistical. The co-dependent relationships and the influence of cancer patients and their close relatives on the mental health of the latter during the six months of the development of hemoblastosis were studied. First identified predictors of mental health of the next of kin hematooncology at different stages of the disease of acute leukemia. New data on the structure and dynamics of the influence of various factors on the mental health of relatives of oncohematological patients through latent variables were obtained. The variants of disadaptation of the closest relatives of oncohematological patients during the first six months of the disease course are indicated. For the first time, a comprehensive approach to the problem of providing medical and psychological care is justified, including the mental health of the closest relatives of oncohematological patients in relation to the psychoemotional status of patients with hemoblastoses. The mechanisms of functioning of relatives of oncohematological patients, which have an impact on their mental health, are revealed, which is of differential importance in correctional and psychotherapeutic practice.Key Words: Relatives of cancer patients; distress; coning strategies; mental health; anxiety; depression; codependent relationships; quality of life; oncology; psychotherapy.