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Exciting perspectives for Translational Myology in the Abstracts of 
the 2018Spring PaduaMuscleDays: Giovanni Salviati Memorial – 
Chapter IV - Abstracts of March 17, 2018 

 

Ugo Carraro (1,2,3) 

(1) Laboratory of Translational Myology, Department of Biomedical Sciences, University of 

Padova; (2) A&C M-C Foundation for Translational Myology, Padova; (3) IRCCS Fondazione 

Ospedale San Camillo, Venezia-Lido, Italy 

This article is distributed under the terms of the Creative Commons Attribution Noncommercial License (CC BY-NC 4.0) which permits 

any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. 

Abstract 

Myologists working in Padua (Italy) were able to continue a half-century tradition of studies of 

skeletal muscles, that started with a research on fever, specifically if and how skeletal muscle 

contribute to it by burning bacterial toxin. Beside main publications in high-impact-factor 

journals by Padua myologists, I hope to convince readers (and myself) of the relevance of the 

editing Basic and Applied Myology (BAM), retitled from 2010 European Journal of 

Translational Myology (EJTM), of the institution of the Interdepartmental Research Center of 

Myology of the University of Padova (CIR-Myo), and of a long series of International 

Conferences organized in Euganei Hills and Padova, that is, the PaduaMuscleDays. The 

2018Spring PaduaMuscleDays (2018SpPMD), were held in Euganei Hills and Padua (Italy), in 

March 14-17, and were dedicated to Giovanni Salviati. The main event of the “Giovanni Salviati 

Memorial”, was held in the Aula Guariento, Accademia Galileiana di Scienze, Lettere ed Arti of 

Padua to honor a beloved friend and excellent scientist 20 years after his premature passing. 

Using the words of Prof. Nicola Rizzuto, we all share his believe that Giovanni “will be 

remembered not only for his talent and originality as a biochemist, but also for his unassuming 

and humanistic personality, a rare quality in highly successful people like Giovanni. The best 

way to remember such a person is to gather pupils and colleagues, who shared with him the 

same scientific interests and ask them to discuss recent advances in their own fields, just as 

Giovanni have liked to do”. Since Giovanni’s friends sent many abstracts still influenced by their 

previous collaboration with him, all the Sessions of the 2018SpPMD reflect both to the research 

aims of Giovanni Salviati and the traditional topics of the PaduaMuscleDays, that is, basics and 

applications of physical, molecular and cellular strategies to maintain or recover functions of 

skeletal muscles. The translational researches summarized in the 2018SpPMD Abstracts are at 

the appropriate high level to attract endorsement of Ethical Committees, the interest of 

International Granting Agencies and approval for publication in top quality international 

journals. The abstracts of the presentations of the March 16, 2018 Padua Muscle Day and those 

of the remaining Posters are listed in this chapter IV. The Author Index of the 2018Spring 

PaduaMuscleDays follows at page 78. 

 

Key Words: Giovanni Salviati, proof of concept, translational myology, PaduaMuscleDays 
Eur J Transl Myol 28 (1): 49-78, 2018 

 



2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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Abstracts of the 2018Spring PaduaMuscleDay, March 17, 2018 

Pain&Mobility, past, present, future: Topics to be 

discussed 

Helmut Kern 

Physiko- und Rheumatherapie, St. Poelten, Austria 

E-mail: helmut@kern-reha.at  

Key words:  Pain&Mobility, past, present, future: Topics to be 

discussed  

An incomplete list of topics follows that should be 

discussed between experts on still open scientific 

question,157-163 such as:  

A - 10 years post RISE 

1. Training parameters on DDM 

2. Improvement of exhausted muscle fibers in DDM by 

cell therapy 

3. Stimulation parameter depending on time from SCI, 

age, magnetic stim, stim during rest, etc  

B - New research for new solutions on old questions 

1. Common new research projects as ageing, pain, 

ostheo-articular-muscular mobility impairments, etc. 

2. Translational research of new topics 

3. New research markers (only from blood, saliva, skin, 

hair samples?) 

C - Working groups on:  

1. nerve regeneration 

2. training-parameters of seniors in different ages 

3. muscle adaptation 

4. connective tissue, cartilage 

157. Kern H, Carraro U, Adami N, et al. Home-based 

Functional Electrical Stimulation (h-b FES) recovers 

permanently denervated muscles in paraplegic patients 

with complete lower motor neuron lesion. Neurorehab 

Neur Rep 2010;24:709-21. 

158. Carraro U, Boncompagni S, Gobbo V, et al. Persistent 

Muscle Fiber Regeneration in Long Term Denervation. 

Past, Present, Future. Eur J Transl Myol 2015 Mar 

11;25:4832. doi: 10.4081/ejtm.2015.4832. eCollection 

2015 Mar 11. Review. 

159. Carraro U, Kern H. Severely Atrophic Human Muscle 

Fibers With Nuclear Misplacement Survive Many Years 

of Permanent Denervation. Eur J Transl Myol. 2016 Jun 

13;26:5894. doi: 10.4081/ejtm.2016.5894. PMID: 

27478559.  

160. Zampieri S, Mosole S, Löfler S, et al. Physical Exercise 

in Aging: Nine Weeks of Leg Press or Electrical 

Stimulation Training in 70 Years Old Sedentary Elderly 

People. Eur J Transl Myol 2015; 25: 237–42. doi: 

10.4081/ejtm. 2015.5374. 

161. Robbins WR, Staats PS, Levine J, et al. Treatment of 

intractable pain with topical large-dose capsaicin: 

preliminary report. Anesthesia and analgesia. 1998 

Mar;86:579-83. PubMed PMID: 9495419. Epub 

1998/03/12. eng. 

162. Derry S, Moore RA. Topical capsaicin (low 

concentration) for chronic neuropathic pain in adults. 

The Cochrane database of systematic reviews. 

2012;9:CD010111. PubMed PMID: 22972149. Epub 

2012/09/14. eng. 

163. Albertin G, Hofer Ch, Zampieri, et al. In complete SCI 

patients, long-term Functional Electrical Stimulation of 

permanent denervated muscles increases epidermis 

thickness. Neurol Res 2018, https://doi.org/10.1080/ 

01616412.2018.1436877. 

***** 

The physiopathologic dialogue between muscle and 

nerve 

Antonio Musarò 

Sapienza University, Rome, Italy 

E-mail: antonio.musaro@uniroma1.it 

Key Words: Aging, ALS, NMJ, PKC, muscle-nerve interplay, 

SOD1G93A 

A crucial system severely affected in several 

neuromuscular diseases is the loss of effective 

connection between muscle and nerve, leading to a 

pathological non-communication between the two 

 
Fig 26. A schematic model depicting the 

physiopathologic interplay between nerve 

and muscle. Alterations in structural, 

physiological and metabolic parameters in 

motor neurons and muscle might act 

synergistically to exacerbate the disease. 

Moreover, morphological and functional 

alterations in muscle (red star) can 

negatively impact motor neuron in a sort of 

"dying-back" process. 

 

mailto:helmut@kern-reha.at
https://doi.org/10.1080/
mailto:antonio.musaro@uniroma1.it


2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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51 

 

tissues. Aging-sarcopenia and several neuromuscular 

diseases, including Amyotrophic Lateral Sclerosis (ALS) 

and muscular dystrophies, are characterized by alteration 

in the functional connection between nerve and muscle, 

creating a sort of short-circuit that impinges the proper 

function between the two systems (Rudolf et al, 2014).164 

Neuromuscular junctions (NMJ) serve as the interface 

between the nervous and skeletal muscular systems, and 

thus they may receive pathophysiological input of both 

pre- and post-synaptic origin (Figure 26). However, 

controversy exists over whether NMJ dismantlement is a 

pathogenic event directly associated with the primary 

defects occurring in motor neurons or whether it occurs 

independently from motor neuron degeneration. To 

address this question, we made use of MLC/SOD1G93A 

transgenic mice (Dobrowolny et al, 2008),165 which 

represent an ideal model to separate the ubiquitous toxic 

effects of mutant SOD1G93A with that of tissue-specific 

effects. We recently provided evidence that increased 

oxidative stress, induced by muscle-specific 

accumulation of mutant SOD1G93A, is causally linked 

to morphological alterations of the neuromuscular 

presynaptic terminals, high turnover rate of 

Acetylcholine Receptor (AChR), and NMJ 

dismantlement (Dobrowolny et al, 2017).166 We then 

disclosed the molecular mechanisms by which oxidative 

stress, mediated by SOD1G93A gene expression, induces 

NMJ dismantlement. Interestingly, we found that muscle 

expression of toxic SOD1G93A gene induces the re-

activation of PKCθ, a serine/threonine kinase implicated 

in the clustering and stability of AChRs during 

development (Lanuza et al, 2001).167 We demonstrated 

that PKCθ selectively colocalizes with AChR in the 

muscle of MLC/SOD1G93A mice. To validate the 

hypothesis that the re-activation of PKCθ expression and 

activity was mechanistically associated with the 

dismantlement of NMJ, we pharmacologically interfered 

with PKCθ activity. We demonstrated that the inhibition 

of PKCθ activity was sufficient to reduce PKCθ - AChR 

co-localization, to restore mitochondrial functionality, to 

rescue the morphological complexity of NMJ, and to 

stabilize AChR turnover (Dobrowolny et al, 2017).166 

These results indicate that increased levels of oxidative 

stress and up-regulation of PKCθ are causally linked to 

NMJ dismantlement and suggest that primary muscle 

defects impact the functional connection between muscle 

and nerve at the level of NMJ. This might represent an 

early pathogenic signature of sarcopenia and 

neuromuscular diseases. In conclusion, our study 

discloses the molecular mechanism that triggers 

functional denervation associated with increased levels 

of oxidative stress within the muscle and suggests 

pharmacological intervention to attenuate muscle 

dysfunction, NMJ loss and eventually disease 

progression, typical of sarcopenia, ALS, and muscle 

diseases,168,169 in a sort of "saving-back" process. 

164. Rudolf R, Khan MM, Labeit S, Deschenes MR. 

Degeneration of neuromuscular junction in age and 

dystrophy. Front Aging Neurosci 2014;6:99. 

165. Dobrowolny G, Aucello M, Rizzuto E, et al. Skeletal 

muscle is a primary target of SOD1G93A-mediated 

toxicity. Cell Metab 2008;8:425-36.  

166. Dobrowolny G, Martini M, Scicchitano BM, et al. Muscle 

Expression of SOD1G93A Triggers the Dismantlement of 

Neuromuscular Junction via PKC-Theta. Antioxid Redox 

Signal 2017 Oct 30. doi: 10.1089/ars.2017.7054. [Epub 

ahead of print]. 

167. Lanuza MA, Garcia N, Santafe M, et al. Pertussis toxin-

sensitive G-protein and protein kinase C activity are 

involved in normal synapse elimination in the neonatal 

rat muscle. J Neurosci Res 2001 63:330-40. 

168. Ortolan P, Zanato R, Coran A, et al. Role of Radiologic 

Imaging in Genetic and Acquired Neuromuscular 

Disorders. Eur J Transl Myol 2015;25:5014. doi: 

10.4081/ejtm.2015.5014. eCollection 2015 Mar 11. 

Review. 

169. Pond A, Marcante A, Zanato R, et al. History, 

Mechanisms and Clinical Value of Fibrillation Analyses 

in Muscle Denervation and Reinnervation by Single Fiber 

Electromyography and Dynamic Echomyography. Eur J 

Transl Myol 2014;24:3297. doi: 

10.4081/ejtm.2014.3297. eCollection 2014 Mar 31. 

***** 

Mobility impairment in low back pain patients and 

its improvement by multi-modal physical therapies 

Nejc Sarabon (a,b,c), Stefan Löfler (c,d), Christian 

Hofer (c), Sandra Zampieri (e), Helmut Kern (c) 

(a) University of Primorska, Faculty of Health Sciences, 

Koper, Slovenia; (b) S2P, Science to Practice Ltd., 

Laboratory for Motor Control and Motor Behaviour, 

Ljubljana, Slovenia; (c) Physiko- und Rheumatherapie, 

St. Poelten, Austria; (d) Department of Physical 

Medicine and Rehabilitation, Wilhelminenspital, 

Vienna, Austria; (d) Interdepartmental Research Center 

of Myology, c/o Department of Biomedical Sciences, 

Padova, Italy 

E-mail: nejc.sarabon@fvz.upr.si 

Keywords: capsaicin, pain, sensation, inflammation. 

Low back pain (LBP) is one of the most common health 

problem of the musculoskeletal system. The aim of our 

clinical study was to test differences in asymptomatic 

participants vs. LBP patients from different perspectives 

of movement function. Additionally, we were interested 

in testing the efficiency of different multi-modal 

regimens of multi-modal therapy, primarily based on 

physical therapy modalities. Altogether, 260 subjects 

were included (30 asymptomatic) and underwent the 

same testing protocol consisting of: (1) questionnaires on 

daily movement function and symptoms, (2) isometric 

strength tests – maximal voluntary force during trunk 

flexion, extension and lateral flexion, (3) flexibility and 

repositioning error of the trunk during trunk bending 

from a standing position, (4) chair rising and stand-up-

mailto:nejc.sarabon@fvz.upr.si


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52 

 

and-go test as functional locomotion tests. Patients 

underwent the same testing protocol before and after 10 

consecutive physiotherapy visits (2 x/week = 5 weeks). 

The patients underwent 3 different multi-modal therapy 

groups that involved the following therapy modalities: 

massage, electrical stimulation, ultrasound, capsaicin-

based patches, and movement exercise. The results 

showed statistically significant differences (p < .05) 

between asymptomatic and LBP subjects in the tested 

parameters. 2-way RM ANOVA revealed significant 

time effects and in majority of cases non-significant time 

x group interaction effects. The outcomes of this study 

add important knowledge to the evidence based practice 

of LBP treatment. Additionally, it sets a foundation for 

future studies of this project aiming at optimization of 

clinical practice. 

170. Gore M, Sadosky A, Stacey BR, et al. The burden of 

chronic low back pain: clinical comorbidities, treatment 

patterns, and health care costs in usual care settings. 

Spine 2012;37:E668-E77. 

171. Hodges P, van den Hoorn W, Dawson A, Cholewicki J. 

Changes in the mechanical properties of the trunk in low 

back pain may be associated with recurrence. J Biomech. 

2009;42:61-6. doi: 10.1016/j.jbiomech.2008. 10.001. 

Epub 2008 Dec 4. 

172. Sarabon N, Löfler S, Jan Cvecka J, Hübl W, Zampieri S. 

Acute effect of different concentrations of cayenne pepper 

cataplasm on sensory-motor functions and serum levels 

of inflammation-related biomarkers in healthy subjects. 

Eur J Transl Myol 2018;28,in press. 

***** 

Histopathological analyses of skin in SCI. New 

results and hopes for h-bFES and beyond 

Giovanna Albertin (a), Helmut Kern (b), Sandra 

Zampieri (b-d), Diego Guidolin (a), Andrea Porzionato 

(a), Barbara Ravara (c-e), Ugo Carraro (c-e) 

(a) Department of Neuroscience, University of Padova, 

Italy; (b) Physiko- und Rheumatherapie, St. Poelten, 

Austria; (c) Department of Biomedical Science, 

University of Padova, Italy; (d) CIR Myo, 

Interdepartmental Research Centre of Myology, 

University of Padova, Italy; (e) A&CM-C Foundation 

for Translational Myology, Padova, Italy 

E-mail: giovanna.albertin@unipd.it  

Key Words: skin thickness, biomarkers, denervated muscle, 

recovery by FES. 

The skin is the body’s heaviest sensory organ, accounting 

for approximately 16% of the body’s weight. Other 

functions are protection from chemical, physical and 

biological insults and maintenance of the internal 

environment.173 Several pathologies are associated to 

skin changes affecting skin cells and other structural 

proteins, thickness of the various epidermal layers, 

inflammatory cells, and amount of water.174 

Simultaneous qualitative and quantitative analysis of 

several components and properties of the skin are 

necessary to understand these disorders and to follow-up 

eventual managements. Our previous studies have shown 

that denervated Quadriceps muscles of patients suffering 

with complete conus and cauda equina lesions were 

rescued by two years of home-based functional electrical 

stimulation (h-bFES) using a new electrical stimulator 

and very large skin electrodes.175-177 Muscle mass, force, 

and structure of the stimulated Quadriceps muscles were 

determined before and after 2 years of h-bFES, using 

Computed Tomography (CT), measurements of knee 

torque during stimulation, and muscle biopsies which 

were analyzed by light and electron microscopy. To 

harvest muscle biopsies the overlying skin was also 

collected and evaluated by histological morphometry of 

H&E and immuno-stained on paraffin-embedded section 

(Figure 27). Analysis of the structural characteristics of 

epidermis, i.e., thickness, morphology of the papillae and 

content of hairs together with some neural and 

inflammatory molecular markers were organized. 

Preliminary results178 are interesting and stimulate 

additional analyses to better describe skin adaptation to 

this peculiar type of electrical stimulation by surface 

electrodes. Our approaches offer also new opportunities 

to study adaptation of the skin to other physical and 

pharmacological therapies based on application of 

 
 
Fig 27. A and B are H&E stains of skin harvested 

from the left thigh before and after h-bFES, 

respectively. Male, aged 48.4 years, 175 cm 

high and 100 kg weight. Th12/L1 SCI. 

Before-FES  biopsy was harvested 3.3 years 

after SCI (A), and after 2.6 years of FES 

(B). Skin thickness increase: +35%, p < 

0.001. 

mailto:giovanna.albertin@unipd.it


2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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rehabilitation managements through the skin, in 

particular for pain relief. 

173. Hunckler J, de Mel A. A current affair: electrotherapy in 

wound healing. J Multidiscip Healthc. 2017:10:179-94. 

174. Kern H, Carraro U, Adami N, et al. Home-based 

functional electrical stimulation rescues permanently 

denervated muscles in paraplegic patients with complete 

lower motor neuron lesion. Neurorehabil Neural Repair 

2010;24:709-721. 

175. Mayr W. Neuromuscular Electrical Stimulation for 

Mobility Support of Elderly. Eur J Transl Myol 

2015;25(4):263-8. doi: 10.4081/ejtm.2015.5605. 

eCollection 2015 Aug 24. 

176. Carraro U, Kern H, Gava P, et al. Biology of Muscle 

Atrophy and of its Recovery by FES in Aging and Mobility 

Impairments: Roots and By-Products. Eur J Transl Myol 

2015;25(4):221-30. doi: 10.4081/ejtm.2015.5272. 

eCollection 2015 Aug 24. Review. 

177. Zampieri S, Mosole S, Löfler S, et al. Physical Exercise 

in Aging: Nine Weeks of Leg Press or Electrical 

Stimulation Training in 70 Years Old Sedentary Elderly 

People. Eur J Transl Myol 2015; 25: 237–242. doi: 

10.4081/ejtm. 2015.5374. 

178. Albertin G, Hofer Ch, Zampieri, et al. In complete SCI 

patients, long-term Functional Electrical Stimulation of 

permanent denervated muscles increases epidermis 

thickness. Neurol Res 2018, https://doi.org/10.1080/ 

01616412.2018.1436877. 

***** 

Discovery of Calcium Entry Units: role in skeletal 

muscle function and disease 

Simona Boncompagni*, Antonio Michelucci*, Laura 

Pietrangelo, Feliciano Protasi 

*Equally contributing Authors 

CeSI-Met, Center for Research on Ageing and 

Translational Medicine; University G. d'Annunzio of 

Chieti-Pescara, Italy 

Email: fprotasi@unich.it  

Key Words: muscle fatigue; sarcoplasmic reticulum (SR); 

skeletal muscle; store-operated Ca2+ entry 

(SOCE) 

Store-operated Ca2+ entry (SOCE) is a ubiquitous 

cellular Ca2+ influx mechanism, first described in non-

excitable cells, that is triggered by depletion of 

intracellular Ca2+ stores (endoplasmic reticulum, ER).179 

A major breakthrough in the field was the identification 

of the two essential molecular players in SOCE: STIM1, 

the Ca2+ sensor in the ER,180 and Orai1, a Ca2+ 

permeable channel in the plasma membrane (PM).181 

Question addressed by this study: SOCE is also well-

documented in skeletal muscle where it limits muscle 

fatigue during repetitive fatiguing stimulation.182,183 Also 

in muscle SOCE is mediated by interactions between 

STIM1 in the SR and Orai1 channels in the PM4. 

However, the precise subcellular location of STIM1-

Orai1 SOCE complexes in skeletal muscle was not yet 

been unequivocally identified. We used a combination of 

ultrastructural and biochemical analyses (electron 

microscopy, immunohistochemistry and confocal 

microscopy, immunogold, and western blot) to compare 

and carefully quantify STIM1 and Orai1 subcellular 

localization in skeletal muscle of adult mice under resting 

conditions and after a single bout of treadmill running 

designed to promote STIM1 aggregation and coupling 

with Orai1. Using this experimental approach, we 

discovered: a) previously unidentified intracellular 

junctions between the sarcoplasmic reticulum (SR) and 

invaginations of the external membrane, i.e. the 

transverse tubules (TTs); b) that this remodeling allows 

for the assembly of the molecular machinery required to 

activate SOCE (i.e. STIM1 and Orai1); c) that muscles 

containing these new SR-TT junctions are more resistant 

to fatigue in  presence of extracellular Ca2+;  d) that TTs 

are more dynamic than SR and retracts from the new 

junctions in a few hours following the exercise protocol. 

We propose that these previously unidentified SR-TT 

junctions function as Ca2+ Entry Units (CEUs) (Figure), 

providing a preferential pathway for rapid reuptake of 

Ca2+ into the SR during repetitive muscle activity.184 

Significance of our study. Our work represents a pioneer 

study that identifies exercise-driven dynamic formation 

of new intracellular structures, an endogenous 

mechanism potentially quite important for the delay of 

muscle fatigue. In addition, as: a) altered SOCE activity 

contributes to muscle dysfunction in ageing and b) 

mutations in STIM1 and Orai1 are linked to Tubular 

Aggregate Myopathy (TAM), our findings may also have 

important implications for a deeper understanding of 

mechanisms involved in muscular dysfunction in 

pathophysiological conditions. 

179. Parekh, A. B. & Penner, R. Store depletion and calcium 

influx. Physiol Rev 1997;77:901-30. 

180. Roos J, DiGregorio PJ, Yeromin AV, et al. STIM1, an 

essential and conserved component of store-operated 

Ca2+ channel function. J Cell Biol 2005;169:435-45. 

181. Vig M,Peinelt C, Beck A, et al. CRACM1 is a plasma 

membrane protein essential for store-operated Ca2+ 

entry. Science 2006;312:1220-3. 

182. Lyfenko AD, Dirksen RT. Differential dependence of 

store-operated and excitation-coupled Ca2+ entry in 

skeletal muscle on STIM1 and Orai1. J Physiol 

2008;586:4815-24. 

183. Wei-Lapierre L, Carrell EM, Boncompagni S, Protasi F, 

Dirksen RT. Orai1-dependent calcium entry promotes 

skeletal muscle growth and limits fatigue. Nat Commun 

2013;4:2805. 

184.  Boncompagni S, Michelucci A, Pietrangelo L, Dirksen 

RT, Protasi F. Exercise-dependent formation of new 

junctions that promote STIM1-Orai1 assembly in skeletal 

muscle. Sci Rep 2017;7:14286. 

***** 

Myofascial force transmission and muscle weakness 

Marco Narici (a), Robert Csapo (b), Usha Sinha (c) 

Shantanu Sinha (d) 

mailto:fprotasi@unich.it
https://www.ncbi.nlm.nih.gov/pubmed/29079778
https://www.ncbi.nlm.nih.gov/pubmed/29079778
https://www.ncbi.nlm.nih.gov/pubmed/29079778


2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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54 

 

(a) Institute of Physiology, Department of Biomedical 

Sciences, University of Padova, Italy; (b) University for 

Health Sciences, Medical Informatics and Technology, 

Institute for Sports Medicine, Alpine Medicine & Health 

(ISAG), Hall, Austria; (c)Physics Department, San 

Diego State University, San Diego, California; (d) 

Muscle Imaging and Modeling Lab, Department of 

Radiology, University of California, San Diego, 

California, USA 

E-mail: marco.narici@unipd.it  

Key Words: skeletal muscle, ageing, sarcopenia, muscle 

quality, extracellular matrix 

Sarcopenia, the age-related loss of muscle mass, affects 

>50% of the population aged 75 years and over and is a 

main cause of impaired physical performance and 

reduced mobility. Amongst the several factors 

contributing to sarcopenia neuroendocrine changes are 

regarded as primary drivers of this condition.185-187 They 

are responsible for -motoneuron- and neuromuscular 

junction (NMJ) degeneration as well as muscle fiber 

denervation that, also fueled by mitochondrial 

dysfunction and oxidative damage,188 leads to loss of 

motor units and muscle weakness. One of the major 

functional characteristics of sarcopenia is the 

disproportionate loss of muscle strength. At the age of 80 

years, the loss of muscle strength is about 4-fold greater 

than that of muscle size.189 This intrinsic muscle 

weakness, also known as a deterioration in ‘muscle 

quality’, has traditionally been attributed to changes in 

muscle fiber type composition, a decrease in fiber 

specific tension, reduced excitation-contraction coupling 

and reduced neural drive. However, new evidence 

suggests that this disproportionate loss of force also 

arises from changes in the extracellular matrix (ECM) 

and of associated proteins,190,191 which in young muscle 

normally contributes to over 50% of muscle force 

output.192 Indeed, as recently reported for rat muscle, 

lateral force transmission is reduced by up to 44% in old 

animals.193 Direct measurements of lateral force 

transmission involve highly invasive surgical procedures 

(tenotomies and myotomies), precluding human studies. 

In this pilot in-vivo study, we tested the hypothesis that 

MR imaging-based indices derived from strain rate (SR) 

tensor maps reflect lateral force transmission (Figure 

28).194 SR tensors were derived from velocity encoded 

magnetic resonance phase-contrast images in nine young 

(28 years) and eight senior (78 years) women. The central 

finding of this study was that the angle enclosed by the 

SR along the fiber (indicative of the principal axis of 

muscle shortening) and the muscle fiber axis was 

significantly smaller in older women (proximal: -19.2%, 

central: -17.6%). Under the assumption that the SR-fiber 

angle would be 0° if force was solely transmitted along 

the fiber, this finding indicates lower lateral transmission 

of force in older compared to younger women. This 

observation seems consistent with the hypothesis, and 

with observations in animal muscle, that a reduction in 

lateral force transmission may contribute to the intrinsic 

muscle weakness of older human muscle. 

185. Narici MV, Maffulli N. Sarcopenia: characteristics, 

mechanisms and functional  significance. Br Med Bull 

2010;95:139-59. doi: 10.1093/bmb/ldq008. 

 
 

Fig 28. Difference in angle enclosed by the fibre SR and the muscle fiber orientation 

 

mailto:marco.narici@unipd.it


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186. Barber L, Scicchitano BM, Musaro A. Molecular and 

Cellular Mechanisms of Muscle Aging and Sarcopenia 

and Effects of Electrical Stimulation in Seniors. Eur J 

Transl Myol 2015;25:231-6. doi: 

187. Power GA, Dalton BH, Gilmore KJ, et al. Maintaining 

Motor Units into Old Age: Running the Final Common 

Pathway. Eur J Transl Myol 2017;27:6597. doi: 

10.4081/ejtm.2017.6597. eCollection 2017 Feb 24. 

188. Protasi F. Mitochondria Association to Calcium Release 

Units is Controlled by Age and Muscle Activity. Eur J 

Transl Myol 2015;25:257-62. doi: 10.4081/ejtm.2015. 

5604. eCollection 2015 Aug 24. 

189. Moore AZ, Caturegli G, Metter EJ, et al. Difference in 

muscle quality over the adult life span and biological  

correlates in the Baltimore Longitudinal Study of Aging. 

J Am Geriatr Soc 2014;62:230-6. doi: 

10.1111/jgs.12653. 

190. Lieber RL, Ward SR. Cellular mechanisms of tissue 

fibrosis. 4. Structural and functional consequences of 

skeletal muscle fibrosis. Am J Physiol Cell Physiol 

2013;305:C241-52. doi: 10.1152/ajpcell.00173.2013. 

191. Zhang C, Gao Y. Effects of aging on the lateral 

transmission of force in rat skeletal muscle. J Biomech 

2014;47:944-8. doi: 10.1016/j.jbiomech.2014.01.026. 

192. Huijing PA, Baan GC, Rebel GT. Non-myotendinous 

force transmission in rat extensor digitorum longus 

muscle. J Exp Biol 1998;201(Pt 5):683-91.  

193. Ramaswamy KS, Palmer ML, van der Meulen JH, et al. 

Lateral transmission of force is impaired in skeletal 

muscles of dystrophic mice and very old rats. J Physiol 

2011;589(Pt5):1195-208. doi: 10.1113/jphysiol.2010. 

201921.  

194. Sinha U, Malis V, Csapo R, et al. 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. Magn Reson 

Med 2015;73:1852-63. doi: 10.1002/mrm.25312. 

***** 

E-T Coupling of muscle trophism modulation 

Alessandra Nori (a), Simone Mosole (a,b), Sandra 

Zampieri (a,b), Sandra Furlan (c), Ugo Carraro (a,d), 

Stefan Löfler (b), Helmut Kern (b,e), Pompeo Volpe (a) 

(a) Department of Biomedical Sciences, University of 

Padova Interdepartmental Research Center of Myolgy 

(CIR-Myo), & IIM, Padova, Italy; (b) Ludwig 

Boltzmann Institute of Electrical Stimulation and 

Physical Rehabilitation, Vienna, Austria; (c) Institute of 

Neuroscience, CNR, Padova, Italy; (d) A&CM-C 

Foundation, Padova, Italy; (e) Institute of Physical 

Medicine and Rehabilitation, Physiko- und 

Rheumatherapie, St. Pölten, Austria. 

E-mail: alessandra.nori@unipd.it  

Key words: Aging, muscle remodeling, electrical stimulation, 

Ca-handling-proteins, NFATc1 

Physical activity plays an important role in preventing 

muscle atrophy in elderly. Calcium cycling and 

activation of specific molecular pathways are essential in 

contraction-induced muscle adaptation. Muscle sections 

and total homogenates were prepared from biopsies 

obtained before and after nine weeks of electrical 

stimulation (ES) on a group of volunteers. NFATc1 

nuclear localization, kinase activation and expression of 

Sarcalumenin, Calsequestrin and sarco/endoplasmic 

reticulum Ca2+-ATPase (Serca) isoforms were 

determined by immunofluorescence and western blot. 

Results support the conclusion that ES is able to 

modulate expression of key Ca-handling proteins 

promoting positive fiber remodeling in sedentary 

seniors.195-199 

195. Boncompagni S, d'Amelio L, Fulle S, et al. Progressive 

disorganization of the excitation-contraction coupling 

apparatus in aging human skeletal muscle as revealed by 

electron microscopy: a possible role in the decline of 

muscle performance. J Gerontol A Biol Sci Med Sci 

2006;61:995-1008. 

196. Salanova M1, Bortoloso E, Schiffl G, et al. Expression 

and regulation of Homer in human skeletal muscle during 

neuromuscular junction adaptation to disuse and 

exercise. FASEB J 2011;25:4312-25.  

197. Ehlers ML, Celona B, Black BL. NFATc1 controls 

skeletal muscle fiber type and is a negative regulator of 

MyoD activity. Cell Rep 2014;8:1639-48. 

198. Zampieri S, Mosole S, Löfler S, et al. Physical Exercise 

in Aging: Nine Weeks of Leg Press or Electrical 

Stimulation Training in 70 Years Old Sedentary Elderly 

People. Eur J Transl Myol 2015;25:237-42. doi: 

10.4081/ejtm.2015.5374. eCollection 2015 Aug 24. 

Review. 

199. Mosole S, Carraro U, Kern H, et al 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. Eur J Transl Myol 2016;26:5972. doi: 

10.4081/ejtm.2016.5972. eCollection 2016 Sep 15. 

***** 

Physiotherapy for knee osteoarthritis: effects on 

lower limb strength, function and gait 

Nathaly Gaudreault 

École de réadaptation, Faculté de médecine et des 

sciences de la santé de l’Université de Sherbrooke, 

Sherbrooke, Quebec, Canada. Centre de recherche du 

centre hospitalier universitaire de Sherbrooke (CR-

CHUS), axe inflammation/douleur, Quebec, Canada. 

Email: Nathaly.Gaudreault@usherbrooke.ca  

Key Words: Knee osteoarthritis, physiotherapy, gait 

mailto:alessandra.nori@unipd.it
mailto:Nathaly.Gaudreault@usherbrooke.ca


2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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56 

 

According clinical guideline recommendations, lower 

limb strengthening exercises are key components of knee 

osteoarthritis (OA) management.200. Exercise has shown 

to have beneficial effects on decreasing symptoms of 

pain and improving physical function in knee OA 

patients. However, its effects on knee biomechanics are 

still unclear. Although changes in knee biomechanics 

during gait have been recently reported following a 

physiotherapy treatment,201 other studies were not 

conclusive.202 Interpreting gait data is challenging due to 

intersubject variability observed in the gait pattern of 

both normal and pathological populations. In this study, 

we investigated the impact of using principal component 

analysis for clustering knee osteoarthritis (OA) patients' 

gait data when studying the effect of a physiotherapy 

treatment focusing on lower limb strengthening 

exercises. 3D knee kinematic and kinetic data were 

recorded during the gait of 29 participants diagnosed with 

knee OA before and after they received 12 weeks of 

physiotherapy treatment. Pain, stiffness, function and 

knee muscle strength were also measured. The 

physiotherapy program was standardized and it was 

mainly oriented towards muscle strengthening and 

stretching exercises, proprioceptive exercises and aerobic 

training. In most gait studies, dynamic joint angles and 

moment data as a function of the gait cycle are presented 

in the form of curves.203 Specific kinematic or kinetic gait 

parameters, such as the mean of peak values, are 

extracted at particular periods of the gait cycle and used 

for group comparison. However, limitations can be 

encountered using this technique. Although human gait 

is a cyclic and repeatable activity, every person has a 

fairly unique gait pattern, leading to intersubject 

variability in curve profiles. Averaging can collapse 

information to the point of removing important 

intersubject variability within a given group, whether 

before or after physiotherapy. In this study, we used 

principal component analysis to extract clusters of knee 

flexion/extension, adduction/abduction and 

internal/external rotation angle and moment data. The 

treatment’s effect was assessed using paired t-tests 

performed before and after clustering the knee kinematic 

data into subgroups. At the end of the physiotherapy 

program, patients had less knee pain, less stiffness and 

felt less disabled; isometric quadriceps and hamstring 

strength was also improved (p<0.05). Except for the knee 

flexion/extension angle, two different clusters (C1 and 

C2) were extracted from the angle and moment data. 

When pre- and post-treatment analyses were performed 

on the clustered data, participants exhibiting a C2 knee 

moment pattern demonstrated a greater first peak flexion 

moment (Figure 29), lower adduction moment impulse 

and smaller rotation angle range post-treatment (p<0.05). 

Pre- and post-treatment comparisons performed on 

unclustered data showed no treatment effect. The results 

of the present study suggest that the proposed 

physiotherapy exercices program is effective to improve 

the clinical status of patients with knee OA. Morevover, 

the results demonstrate that the effects of exercices on 

knee biomechanics may be masked or underestimated if 

the gait data are not clustered into more homogeneous 

subgroups when performing pre- and post-treatment 

comparisons. 

200. Zhang W, Moskowitz RW, et al. OARSI recommendations 

for the management of hip and knee osteoarthritis, part I: 

critical appraisal of existing treatment guidelines and 

systematic review of current research evidence. 

Osteoarthritis Cartilage 2007;15,981-1000. 

201. Turcot K, Aissaoui R, Boivin K, et al. The responsiveness 

of three-dimensional knee accelerations used as an 

estimation of knee instability and loading transmission 

 
Fig 29.  Sagittal plane knee moments curve profils, clustered data (C1 and C2) as a function of normalize gait cycle. 

When pre- and post-treatment analyses were performed using the clustered data, a significantly greater first 

peak flexion moment (M1) was observed for patients exhibiting C2 patterns (p<0.05). 

 



2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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57 

 

during gait in osteoarthritis patient's follow-up. 

Osteoarthritis Cartilage 2009;17,213-9. 

202. Thorstensson CA, Henriksson M, von Porat A, et al. The 

effect of eight weeks of exercise on knee adduction 

moment in early knee osteoarthritis--a pilot study. 

Osteoarthritis Cartilage 2007;15:1163-70. 

203. Winter DA. The biomechanics and motor control of 

human gait., Waterloo, ON, University of Waterloo 

Press. 1988. 

***** 

FES-cycling at the 1st CYBATHLON competition 

Benoît Sijobert (a), Charles Fattal (b), Christine 
Azevedo Coste (a) 

(a) INRIA-LIRMM, Montpellier; (b)  CRF La 

Châtaigneraie, France 

E-mail: christine.azevedo@inria.fr 

Key Words: adaptive control of movements, gait and cycling 

assistance, Functional Electrical Stimulation, 

FES leisure cycling for SCI, CYBATHLON 

Functional Electrical Stimulation (FES) can elicit 

muscular contraction and restore motor function in 

paralyzed limbs. FES can assist cycling for training 

purposes mainly in clinical environments using cycle 

ergometers. Different events have promoted FES-cycling 

as a recreational practice. In 2016, twelve teams 

participated in the first Cybathlon competition in the 

FES-cycling discipline for persons with motor-complete 

spinal cord injury. Different approaches have been 

considered by the different participating teams.204 The 

objective of the race was to travel 742 m in less than 8 

minutes. 7 pilots were able to cover the total distance in 

5.5mn on average.205 This communication intends to 

comment the results and performances during this 

competition and discuss the scientific challenges to 

improve FES-assisted cycling applications. Taking the 

example of the experience of Freewheels team we will 

also discuss the experience of participating in such an 

event.206 Online modulation of functional electrical 

stimulation (FES) parameters is necessary to adapt 

timings and levels of muscle activations. This requires 

information on the system state evolution and involves 

the design of robust closed-loop control approaches. 

Open-loop control should be limited to non functional 

situations and to motion control not requiring 

reproducibility or adaptability (e.g facing fatigue 

occurrence or external changes). Voluntary actions of the 

user through his/her healthy limbs or residual control of 

his/her deficient limbs should also be considered and 

integrated in the global functional task control as well 

and not only under the form of external disturbances. Our 

team has been working for several years on the 

observation of voluntary movements through artificial 

sensors in order to extract useful information about the 

on-going postural task.207-209 Indeed, depending on the 

considered function to be restored (posture, gait, 

grasping, cycling), different information can be extracted 

from embedded sensors, going from discrete events to 

continuous measurements of variables (joint angles, gait 

evolution…). As illustrated in Figure 30, this method 

involves combining a natural controller (i.e. the subject 

with his/her disability and his/her partial voluntary 

control) with an artificial FES-based controller fed with 

information from one or multiple sensors located on the 

subject. We will discuss possible improvements of FES-

cycling assistance, using at a deeper level sensors 

embedded either on the tricycle or on the user, paving the 

way to adaptive assistive solutions 

204. Azevedo Coste C, Bergeron V, Berkelmans R, et al. 

Comparison of strategies and performance of functional 

electrical stimulation cycling in spinal cord injury pilots 

for competition in the first ever CYBATHLON. Eur J 

Transl Myol 2017;27:251-4.  

205. Azevedo Coste C, Wolf P. FES-cycling at Cybathlon 

2016: Overview on Teams and Results. Artificial Organs 

2018, in press. 

206. Sijobert B, Fattal C, Daubigney A, Azevedo Coste C. 

Participation to the first Cybathlon: an overview of the 

FREEWHEELS team FES-cycling solution. Eur J Transl 

Myol 2017;27:265-71. 

207. Azevedo Coste C, Jovic J, Pissard-Gibollet R, Froger J. 

Continuous gait cycle index estimation for electrical 

stimulation assisted foot drop correction. J Neuroeng 

Rehab 2014;11:118-28. 

208. Jovic J, Azevedo Coste C, Fraisse P, et al. Coordinating 

Upper and Lower Body During FES-Assisted Transfers 

in Persons With Spinal Cord Injury in Order to Reduce 

Arm Support. Neuromodulation 2015;18:736-42. 

209. Azevedo Coste C, Sijobert B, Geny C, et al. The potential 

of inertial sensors in posture, gait and cycling FES-

assistance. ICNR: Int Conf on Neurorehabilitation 2016. 

***** 

Functional electrical stimulation for ageing laryngeal 

muscles 

 
 
Fig 30. Illustrative examples of a closed-loop approach of 

FES-assisted postural tasks restoration. 

 

 

mailto:christine.azevedo@inria.fr


2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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58 

 

Markus Gugatschka (a), Jonathan C. Jarvis (b), Justin 

Perkins (c), Claus Gerstenberger (a), Michael 

Karbiener (a) 

(a) Dept. of Phoniatrics, Medical University Graz, 

Austria; (b) School of Sport and Exercise Science, 

Liverpool John Moores University, Liverpool, United 

Kingdom; (c) Department of Veterinary Clinical 

Sciences, Royal Veterinary College, North Mymms, UK 

E-mail: markus.gugatschka@medunigraz.at  

Key words: Evaluation, surface FES, voice rejuvenation 

The percentage of elderly people is increasing steadily 

during the last decades in most Western societies and 

large parts of Asia which is accompanied by a steady 

increase of age related diseases. These changes do not 

spare the larynx. Weakening of the voice has long been 

neglected, but gained consideration more recently, as 

vocal endurance is required in many professions up to 

higher ages. A large retrospective study identified vocal 

fold (VF) atrophy as the most prevalent finding. The 

noticeable glottal gap and VF bowing are the most 

prominent video-laryngoscopic findings in these 

patients, and are related to the atrophy of the 

thyroarytenoid muscle (TAM). In a recent trial we could 

demonstrate that functional electrical stimulation (FES) 

of the afferent nerves with specific parameters led to a 

muscle fiber hypertrophy.210 In subsequent trials we 

explored not only different stimulation patterns, but also 

different anatomical locations and outcome parameters 

such as 3D-volumetry.211 Aged sheep were used as 

animal model. FES was delivered in predesigned training 

patterns unilaterally at the common trunk of the recurrent 

laryngeal nerve, as well as at more distal ends of the same 

nerve (mere adduction fibers). After a maximum time of 

eleven weeks permanent stimulation we observed 

significant increases of mean fiber diameters and 

volumes of the TAM using different stimulation patterns. 

Noticeably, we did not elicit a fiber type change with any 

regime, as proved with RT-qPCR. 

FES is possible new treatment option to counteract 

laryngeal sarcopenia. Further work will comprise 

identification of optimized patterns in terms of training- 

and current-efficacy. 

210. Karbiener M, Jarvis JC, Perkins JD, et al. Reversing 

Age Related Changes of the Laryngeal Muscles by 

Chronic Electrostimulation of the Recurrent Laryngeal 

Nerve. PLoS One. 2016;11(11):e0167367 

211. Gerstenberger C, Döllinger M, Kniesburges S, et al. 

Phonation Analysis Combined with 3D Reconstruction 

of the Thyroarytenoid Muscle in Aged Ovine Ex Vivo 

Larynx Models. J Voice 2017;pii: S0892-

1997(17)30215-1. 

***** 

Selective surface stimulation of denervated muscles 

in the larynx 

Berit Schneider-Stickler (a), Matthias Leonhard (b), 

Winfried Mayr (b), 

(a) Department of Otorhinolarangology/Devision of 

Phoniatrics-Logopedics Vienna; (b) Medical Center for 

Medical Physics and Biomedical Engineering; 

University Vienna, Austria 

E-mail: berit.schneider-stickler@meduniwien.ac.at, 

matthias.leonhard@meduniwien.ac.at 

Keywords: unilateral vocal fold paralysis, electrotherapy, 

electrical stimulation, hoarseness 

The laryngeal innervation bears the risk of unilateral and 

bilateral vocal fold paralysis due to trauma, injury during 

surgery, and inflammation. Vocal fold paralysis is a 

pathological motion impairment of the vocal fold, mostly 

caused by damage of the N. vagus or the N. laryngeus 

recurrens. Bilateral vocal fold paralysis usually causes 

dyspnea with inspiratory stridor and requires immediate 

   
 

 
Fig 31. Upper left panel: Uni-polar electrode 

configuration.  
 Upper right panel: Two electrodes (same as 

bipolar set-up) are places above the larynx 

and two indifferent electrodes on the 

shoulder blades (large self-adhesive 

electrodes with 13 x 8 cm). 
 Lower panel: The principle of selective 

stimulation of denervated muscle utilizing a 

long exponentially progressive current 

form. The strength duration curves for 

normally innervated muscle, denervated 

muscle, and intact sensory nerves are 

illustrated. Selective stimulation of the 

denervated muscle without recruitment of 

either normally innervated muscle or the 

sensory axons is possible (shaded area). 

 

mailto:markus.gugatschka@medunigraz.at
mailto:berit.schneider-stickler@meduniwien.ac.at
mailto:matthias.leonhard@meduniwien.ac.at


2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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59 

 

an intervention to stabilize the airway. The unilateral 

vocal fold paralysis (ULVP) causes dysphonia, thus 

patients are in need for therapy to improve the voice 

quality. In temporary ULVP, direct superficial electrical 

muscle stimulation (EMS) can be used for activation of 

intrinsic laryngeal muscles as an alternative to 

conservative voice therapy.212-217 The EMS should 

promote the regeneration without negatively interfering 

with normal regeneration processes. Little is known 

about the effect of electric current on the larynx. In this 

study, 30 patients with ULVP (20 male/10 female, 

average age 57,4 y) underwent superficial electrical 

muscle stimulation (EMS) with flexible endoscopic 

laryngoscopy (Figure 31). Criteria for a selective 

laryngeal activation are an ab- or adduction of the 

immobile vocal fold. In 10/30 patients a selective 

laryngeal activity could be achieved using conventional 

stimulation parameters. The other patients did not present 

any selective vocal fold movement or the test had to be 

ended prematurely due to nonselective reaction (activity 

of the strap muscles, unintentional swallowing, pain) 

before a selective laryngeal activity could be achieved. 

Nevertheless, the superficial electrical muscle 

stimulation can be regarded as a therapy alternative to 

only “wait and see”-strategy and a sole voice therapy in 

early treatment of ULVP. In future studies, position and 

size of electrodes and the stimulation parameters need to 

be improved in order to reach a reliable and effective 

selective laryngeal stimulation. 

212. Hirano M, Kirchner J, Bless D. Neurolaryngology recent 

advances. SPG 1986. 

213. Sulica L, Blitzer A, Vocal Fold Paralysis, Springer 2006. 

214.  Kruse E. Die Reizstrombeh.andlung als integraler 

Bestandteil der logopadischen Stimmtherapie Sprache 

Stimme Gehör 1989;13:64-70. 

215. Mayr W, Hofer C, Bijak M, et al. Functional Electrical 

Stimulation (FES) of Denervated Muscles : Existing and 

Prospective Technological Solutions. Eur J Transl Myol 

2002;12:1–4. 

216. Martin F, Witt TN. Elektrodiagnostische und 

histometrische Untersuchungen über den Einfluß von 

Reizstrom auf die Atrophie der denervierten 

Kehlkopfmuskulatur im Tierexperiment. Laryngol. 

Rhinol. Otol (Stuttg) 1983;62. 

217. Schleier E, Streubel H-G. Beziehungen zwischen 

diagnostischer Aussage und therapeutischem Ergebnis 

bei Rekurrenslähmung. Folia phoniat 1980;32:323-33. 

***** 

Reinnervation is not prevented by surface electrical 

stimulation (ES): clinical experience of 17 patients 

with facial nerve paralysis 

Gerd Fabian Volk (a), Wiebke Puls (a), Wengelawit 

Misikire (a), Anja Greiner (a), Jonathan C. Jarvis (b), 

Orlando Guntinas-Lichius (a) 

(a) ENT-Department and Facial Nerve Center Jena, 

Jena University Hospital, Jena, Germany; (b) School of 

Sport and Exercise Sciences, Liverpool John Moores 

University, Liverpool, United Kingdom 

E-mail: fabian.volk@med.uni-jena.de  

Keywords: Electrostimulation; Reinnervation; Facial 

Paralysis; Facial Pacing; Surface electrodes; 

triangular pulses 

Facial nerve paralysis as a complete peripheral nerve 

injury results in neuromuscular atrophy or in a 

combination of muscle atrophy and aberrant 

reinnervation of facial muscles. The symptoms include 

significant aesthetic, functional and often life-altering 

consequences. Facial pacing systems show controversial 

but promising results to treat facial paralysis.218 Electrical 

stimulation (ES) can increase the mass and force 

generation of denervated lower extremity muscles.219 

Nevertheless, there is an ongoing discussion, whether the 

potential benefit of ES to maintain the muscle might be 

compromised by reducing the chance of reinnervation. 

The few studies published show no clear benefit for ES 

in the face.220 We present data after a new regime of 

home-based stimulation. Before analysing the potential 

positive effects of optimal ES, negative effects have to be 

excluded. From April 2016 till November 2017 17 

patients (11 women) with an age between 18 and 78 

years, with a complete denervation of one facial nerve 

(caused by benign tumor: 8, malign tumor: 4, idiopathic: 

1) confirmed by needle electromyography, have received 

home-based ES devices for exponential impulses training 

to induce contractions of the denervated muscles of the 

mid and lower face (Figure 32). These 17 patients have 

performed ES-training for on average 12months (min. 

4months, max. 21months). The average amplitude was 

14mA (min. 6mA, max. 25mA), the average phase-

duration of the biphasic triangular impulses was 100ms 

(min. 30ms, max. 500ms), the frequency 0.9Hz. Out of 

this heterogeneous group 7 patients got a Hypoglossal-

Facial-Jump-Anastomosis (HFJA), a surgical procedure 

to allow axons from the hypoglossal nerve to reinnervate 

the facial muscles using a nerve-bypass. While waiting 

 

 
 

Fig 32. Placement of two 6x4cm surface 

electrodes on the cheek of a patient 

suffering with postoperative complete 

denervation of the left facial muscles. 

mailto:fabian.volk@med.uni-jena.de


2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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60 

 

for reinnervation, these patients performed ES-training 

for on average 12 months (min. 9 months, max. 18 

months). The average amplitude was 14mA (min. 10mA, 

max. 25mA), the average phase-duration of the biphasic 

triangular impulses was 100ms (min. 70ms, max. 

500ms), the frequency 0.9Hz. Most patients performed 

the training twice a day for 10 minutes and on 5 days a 

week according to their diary. Reinnervation was 

confirmed by volitional movements and by needle EMG 

of the formerly paralysed muscles. 

None of our patients had to stop the ES training due to 

negative side-effects. In 6 patients there were first signs 

of reinnervation before beginning the ES-training. With 

ES, 5 of them showed a progressive reinnervation (more 

muscles innervated and stronger EMG activity per 

muscle). Only one had no improvement of the EMG. 

Reinnervation was observed in 7 patients that had no sign 

of innervation at the start of the training. In 1 patient, 

even 12 month after denervation, no signs of 

reinnervation were detected and in 3 patients, the follow 

up time without reinnervation is less than 12 month. 

Therefore these patients are excluded for our 

retrospective study. So, reinnervation or improvement of 

reinnervation was observed in 85% (12 from 14 patients 

with sufficient follow up) despite electrostimulation. On 

average, it took 8 months after the initial denervating 

infection or surgery until first signs of reinnervation 

could be detected by visual inspection and needle-EMG. 

In the sub-group of patients that received a HFJA, 

reinnervation was observed in 4 of the 7 patients. In one 

patient there are no signs of reinnervation yet due to the 

short period since the HFJA (surgery November 2017). 

Therefore this patient was excluded for our retrospective 

study of the 6 patients reinnervated within 6 months. In 

another patient it took 4 years for stable reinnervation 

with sufficient volitional movements. The defect in the 

cerebellum after extirpation of an astrocytoma could 

explain this unusual long reinnervation time. Two 

patients had first signs of reinnervation before they began 

the ES-training. In the follow up an improvement of 

reinnervation (more muscles innervated and stronger 

EMG activity per muscle) was shown. So reinnervation 

and improvement of reinnervation was observed in 100% 

of the patients with sufficient follow up, despite 

electrostimulation. In conclusions, surface stimulation of 

facial muscles with long triangular biphasic pulses did 

not prevent reinnervation in this series. Most of our 

patients reported a comfortable home based ES training 

recruiting visible muscle response, an improvement in 

facial symmetry and tone, and later on also in muscle 

movement. In the future, these subjective reports have to 

be quantified and examined in detail to differentiate the 

benefit of ES and reinnervation. Seeing no hint of severe 

side effects, facial pacing should be further investigated 

to prove therapeutic effects like reduction of muscle 

atrophy.221 

218. Gittins J, Martin K, Sheldrick J, et al. Electrical 

stimulation as a therapeutic option to improve eyelid 

function in chronic facial nerve disorders. Investig 

Ophthalmol Vis Sci 1999;40:547–54. 

219. Mayr W, Hofer C, Bijak M, et al. Functional Electrical 

Stimulation (FES) of Denervated Muscles : Existing and 

Prospective Technological Solutions. Eur J Transl Myol 

2002;12:1–4. 

220. Fargher KA, Coulson SE. Effectiveness of electrical 

stimulation for rehabilitation of facial nerve paralysis. 

Physical Therapy Reviews 2017. DOI: 

10.1080/10833196.2017.1368967 

221. Volk GF, Sauer M,  Pohlmann M, Guntinas-Lichius O. 

Reference values for dynamic facial muscle 

ultrasonography in adults. Muscle Nerve. 2014;50:348-

57. doi: 10.1002/mus.24204. 

***** 

Opposite effects of tumor-derived cytokines and 

mechanical stimulation on muscle stem cell activity 

and muscle homeostasis 

Alexandra Baccam (a,b,e), Alexandra Benoni (a), Ara 

Parlakian (a), Medhi Hassani (a,b,e), Martina Ramella 

(c), Francesca Boccafoschi (c), Athanassia Sotiropoulos 

(d), Zhigang Xue (b), Viviana Moresi (a), Zhenlin Li (b), 

Sergio Adamo (a), Dario Coletti (a,b,e) 

(a) Sapienza University of Rome, Italy; (b) Sorbonne 

University, France; (c) Eastern Piedmont University, 

Italy; (d) Institut Cochin Paris, France; (e) 

Interuniversity Institute of Myology 

Email:  dario.coletti@uniroma1.it , 

dario.coletti@upmc.fr  

Key Words: skeletal muscle differentiation; myotube 

atrophy; mechanical stimulation; myokines; 

tumor-derived factors 

Cancer cachexia is a muscle wasting syndrome, 

characterized by muscle fiber atrophy and hampered 

satellite cell (SC) myogenic potential, ultimately leading 

to morbidity, lowered quality of life, and death. Exercise 

training improves quality of life and survival of cancer 

patients and its beneficial effects can be mimicked by 

wheel running in mice.222,223 In an animal model of 

cancer cachexia we demonstrated that wheel running 

counteracts cachexia by releasing the autophagic flux and 

by lowering Pax7 expression, which blocks SC myogenic 

progression.224,225. Exercise pleiotropic effects include 

the alteration of circulating factors in favor of an anti-

inflammatory environment and the activation of 

mechanotransduction pathways in muscle cells. Serum 

response factor (SRF) is a transcription factor of pivotal 

importance for muscle homeostasis, which is activated 

with its co-factor MRTF by mechanotransduction in a 

way dependent on actin polymerization.226 Our goal was 

to assess whether mechanotransduction per se is 

sufficient to elicit exercise effects in the presence of pro-

cachectic factors of tumor origin and to characterize the 

mechano-transduction signals involved. We used C26 

tumor-bearing mice, in the absence or presence of wheel 

running, and mixed cultures of C2C12 myotubes and 

mailto:dario.coletti@uniroma1.it
mailto:dario.coletti@upmc.fr


2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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61 

 

myoblasts treated with C26 conditioned medium (CM) in 

the absence or presence of cyclic stretch to mimic the 

mechanical stimulation occurring upon exercise (Figure 

33). In vivo both SRF expression and activity were 

differentially modulated by the C26 tumor, i.e. by 

humoral factors, and by exercise. In vitro we showed that 

CM had a negative effect on muscle cell cultures, both in 

terms of myotube atrophy and of myoblast recruitment 

and fusion, and that these effects were counteracted by 

cyclic stretch. We showed that CM repressed SRF-

MRTF transcriptional activity, while mechanical stretch 

rescued their transcriptional activity; in addition, loss of 

function experiments demonstrated that SRF was 

necessary to mediate the beneficial effects of mechanical 

stimulation on muscle cells. At least part of the observed 

effects was mediated by the balance of pro- and anti-

myogenic factor, such as IL-4 and members of the 

TGFbeta superfamily. in conclusion, we propose that the 

positive effects of exercise on cancer patients and mice 

may be specifically due to a mechanical response of 

muscle fibers affecting the secretion of myokines. 

222. He W, Berardi E, Cardillo VM, et al. NF-kB dependent 

Pax7 deregulation in the muscle microenvironment 

promotes wasting in cancer cachexia. J Clin Invest, 

2013;123:4821-35. 

223. Coletti D, Adamo S, Moresi V. How much a mouse is 

willing to run: having a hard time measuring spontaneous 

physical activity in different mouse sub-strains. Eur J 

Transl Myol 2017; 27(1):67-70. 

224. Pigna E, Berardi E, Aulino P, et al. Aerobic Exercise and 

Pharmacological Treatments Counteract Cachexia by 

Modulating Autophagy in Colon Cancer. Sci Rep 

2016;6:26991. 

225. Coletti D, Aulino P, Pigna E, et al. Spontaneous physical 

activity downregulates Pax7 in cancer cachexia. Stem 

Cells Int. 2016; 2016:6729268. 

226. Coletti D, Daou N, Hassani M, et al. Serum Response 

Factor in muscle tissues: from development to ageing. 

Eur J Transl Myol. 2016; 26(2):6008. 

***** 

Seeking economy of charge injection in activation of 

motor nerves 

Steffen Eickhoff (a),Martin Schmoll (a,b), Jonathan C 

Jarvis (a) 

(a) School of Sport and Exercise Science, Liverpool 

John Moores University, UK; (b) Center for Medical 

Physics and Biomedical Engineering, Medical 

University Vienna, Austria 

E-mail: J.C.Jarvis@ljmu.ac.uk  

Key Words: neural activation, nerve, pulse shaping, 

biphasic pulse, depolarisation, 

How does one choose a pattern of electrical stimulation 

for therapeutic effect? Often there is a useful guide from 

normal physiology, and many therapeutic strategies try to 

mimic or replace a natural activation pattern.227 Another 

strategy is to try to generate a numerical model of the 

excitable tissue to be stimulated so that trials can be 

achieved in silico.228 Many optimised activation 

strategies are based on the results of such simulations. 

They have been used to identify activating pulses that 

best answer the requirement of charge balance to 

minimize damage to electrode and tissue, and the 

requirement to activate nerve tissue with minimal use of 

energy. This latter requirement is critical to improve the 

lifetime of implantable neuromodulation devices. We 

have tested some of the conclusions of studies that have 

investigated the charge efficiency of activation both in 

numerical models and in nerve-muscle preparations.229-

234 We have used the simple experimental model of a 

single motor nerve trunk activated by two electrodes 

 
Fig 33. Tumor cell conditioned medium (CM) induces myotube atrophy and hampers myogenesis. 

Immunofluorescence analysis for MHC (red) on mixed cultures consisting of nascent myotubes and myoblasts 

(nuclei are counterstained in blue), following 4 d of culture (left panel). After additional 2 d of culture in 

differentiation medium (2% HS, Horse Serum) myotube size increases (central panel), while in the presence 

of C26 tumor cell conditioned medium (2% HS + 20% CM) the myotubes appear atrophic and no additional 

nuclei are present in the myotubes in respect to 4 d of culture. This experimental model to test the effect of 

tumor-derived factors on muscle cells, can be combined with mechanical stimulation when the cells are 

cultured in the FlexCell apparatus. 

mailto:J.C.Jarvis@ljmu.ac.uk


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placed near to the nerve (common peroneal in rats). The 

degree of activation has been monitored indirectly by 

measuring the isometric force of the extensor digitorum 

longus muscle because it has discrete proximal and distal 

tendons and can thus be mechanically isolated between a 

proximal clamp and a distal load sensor. Our latest results 

show that the relative benefit of Gaussian or sinusoidal 

shaping of biphasic pulses over rectangular shaping 

reduces with phase width (Figure 34). We will also 

present new data on the complex recruitment curves that 

are generated when the range of phase width and 

amplitude is extended, adding electromyographic 

recordings from the activated muscle to try to understand 

the effect of the primary, secondary and tertiary 

transitions in electrode voltage in a biphasic pulse. Such 

fine differences are important when designing low 

energy implanted stimulators such as may be used in 

retinal stimulation or brain stimulation or activation of 

fine autonomic nerves. 

227.  Henneman E, Somjen G, Carpenter DO. Excitability and 

Inhibitibility of Motoneurons of Different Sizes. J 

Neurophysiol 1965:28:599–620, . 

228. Wongsarnpigoon A, Woock JP, Grill WM. Efficiency 

Analysis of Waveform Shape for Electrical Excitation of 

Nerve Fibers. IEEE Trans Neural Syst Rehabil Eng, 

2010;18:319–28. 

229.  Shepherd RK, Javel E. Electrical stimulation of the 

auditory nerve: II. Effect of stimulus waveshape on single 

fibre response properties. Hearing Research 

1999;130:171–188. 

230. McKay CM, Henshall KR. The perceptual effects of 

interphase gap duration in cochlear implant stimulation. 

Hearing Research 2003;181: 94–9. 

231. Carlyon RP, van Wieringen A, Deeks JM, et al. Effect of 

inter-phase gap on the sensitivity of cochlear implant 

users to electrical stimulation. Hearing Research 

2005;205:210–24.  

232. Prado-Guitierrez P, Fewster LM, Heasman JM, et al. 

Effect of interphase gap and pulse duration on 

electrically evoked potentials is correlated with auditory 

nerve survival. Hear Res 2006;215: 47–55, 

233. Weitz AC , Behrend MR, Ahuja AK, er al. Interphase gap 

as a means to reduce electrical stimulation thresholds for 

epiretinal prostheses. J Neural Eng 2014;11:016007. 

234. Macherey O, van Wieringen A, Carlyon RP, et al. 

Asymmetric Pulses in Cochlear Implants: Effects of Pulse 

Shape, Polarity, and Rate. J Assoc Res Otolaryngol 

2006;7:253–66. 

***** 

Enhanced muscle coordination by transcutaneous 

spinal cord stimulation in brain injured people 

Thordur Helgason (a,b), Vilborg Gudmundsdottir (b), 

Gigja Magnusdottir (b), Rún Friðriksdóttir (a), Margrét 

Sól Ragnarsdóttir (a), Belinda Chenery (c), Gudbjorg 

Ludwigsdottir (b) 

(a) Reykjavik University, Iceland. (b) Landspitali – 

University Hospital, Reykjavik, Iceland. (c) 

University of Iceland 

E-mail: thordur@landspitali.is 

Key Words:  Transcutaneous spinal cord stimulation, 

treatment of spasticity, brain damage, EMG 

Transcutaneous spinal cord stimulation (tSCS) is used in 

our clinic as a therapy option as it has been shown to 

abbreviate spasticity in lower limbs in people with 

incomplete spinal cord injury (SCI) people.235,236 This is 

thought to be inhibiting effects of action potentials 

entering the neural network of the spine where the motor 

neuron for the muscle in question exits.237,238 By 

stimulating the posterior roots of sensory fibres this can 

be reached without stimulating the motor neuron of the 

same muscle. The sensory fibres have bigger diameter 

and therefore lower stimulation threshold compared to 

the motor neurons. The question arises how the tSCS 

influences the spinal cord circuitry. In order to 

investigate on that we looked at EMG in pendulum test 

and in Achilles tendon test by brain injured people. In this 

work four hemiplegic subjects with severe spasm are 

stimulated in the lumbar area, at the height of Th11-

Th12. One man (67) with brain damage after falling, a 

woman (48) after stroke, a woman (72) with brain tumour 

and a woman (68) with a bleeding aneurism. An 

assessment of the spasm was made in the morning 

followed by a 30 minutes posterior root sensory fibres 

stimulating treatment with 50 Hz with no EMG responses 

 
 

Fig 34. Recruitment Curve EDL Muscle. Force 

developed by the extensor digitorum longus 

(EDL) muscle following a biphasic 

rectangular stimulation (phase width 1ms) 

of the Common Peroneal Nerve. The 

Common Peroneal Nerve was isolated from 

the central nervous system. Insert a) EMG 

signal recorded on the EDL muscle at full 

muscle recruitment (first force plateau 400-

1300µA). Insert b) EMG signal recorded on 

the EDL muscle at double muscle activation 

(second force plateau 2000-2700µA) 

 

mailto:thordur@landspitali.is


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63 

 

of the corresponding muscles (sub motor threshold). The 

assessment consist of EMG measurements and 

evaluation of passive and active movements.236 

Stimulating electrodes where placed on the surface on the 

skin at the back and two big different electrodes where 

placed on the abdomen just beside the umbilicus.  

Immediately after the treatment a second assessment was 

made and a third one four to six hours later. Changes in 

the reflex responses of the muscles are recorded.  EMG 

activity during Wattenberg pendulum test and during 

Achilles tendon stretch test is changed towards a clear 

division between the antagonistic muscles. Plantar and 

dorsiflexion of the foot is done easier. EMG shows less 

co-contractions and therefore les effort for the 

movement. Three out of four subject report easier 

movements. The EMG recordings show clear changes 

towards coordination in the responses of the thigh 

muscles after tSCS treatment for 30 minutes showing. 

This is detected both by the Wartenberg pendulum test 

and by the Achilles tendon test. This is in line with the 

results obtained by SCI people. In the SCI group the 

reduction of spasticity was though obvious whereas in the 

brain injured group the effects is not as clear. The effects 

of tSCS on the spinal neural network has to be further 

investigated. 

235. Hofstoetter US; McKay, WB; Tansey KE; et al. 

Modification of Spasticity by Transcutaneous Spinal 

Cord Stimulation in Individuals with Incomplete Spinal 

Cord Injury. J Spinal Cord Med 2014;37:202-11. doi: 

10.1179/2045772313Y.0000000149. Epub 2013 Nov 26.. 

236. Vargas Luna JL; Gudfinnsdottir HK.; Magnusdottir G; et 

al:  Effects fo Sustained Electrical Stimulation on 

Spasticity Assessed by the Pendulum Test. Curren 

Directions in Biomedical Engineering 2016;2:405-7. 

237. Dimitrijevic MR: Restorative Neurology of Spinal Cord 

Injury. New York: Oxford University Press, 2012. 

238. Levins A, Moritz CT. : Therapeutic Stimulation for 

Restoration of Function after Spinal Cord Injury. 

Physiology 2017;32:391-8. . doi: 10.1152/physiol. 

00010.2017. 

***** 

Effect of ageing on the myosin heavy chain 

composition of the human sternocleidomastoid 

muscle 

Marija Meznaric (a), Ida Eržen (a), Petr Karen (b), 

Erika Cvetko (a) 

(a) Institute of Anatomy, Faculty of Medicine, 

University of Ljubljana, Ljubljana, Slovenia; (b) 

Department of Biomathematics, Institute of Physiology, 

Academy of Sciences of the Czech Republic, Czech 

Republic 

E-mail: marija.meznaric@mf.uni-lj.si  

Key words: Human, Ageing, Sternocleidomastoid muscle, 

Immunohistochemistry, Myosin heavy chain 

isoforms, Muscle fiber types 

The underlying cause of sarcopenia, or the age-related 

loss of muscle mass, in limb muscles appears to be slowly 

progressive denervation due to age related motor neuron 

loss in the lumbar and cervical  spinal cord with 

incomplete reinnervation of muscle fibres.239,240 In 

ageing limb muscles morphological studies  

demonstrated a slower phenotype,241 in accordance with 

the preferential loss of fast motor neurons.242 

Concomitant muscle fiber atrophy, mainly of type 2,243 is 

thought to be related to disuse. Thus disuse seems to act 

in the opposite direction to partial denervation, that either 

determines a shift towards the faster phenotype or no 

change in the phenotype.243 The Sternocleidomastoid 

(SCM) muscle is an interesting skeletal muscle to 

investigate since SCM activity is essential for everyday 

tasks and its activity during life is relatively unchanged, 

avoiding to some extent inactivity-related atrophy to 

which the limb muscles are prone. By analyzing 

immunohistochemically myosin heavy chain (MyHC) 

composition we have demonstrated that a similar fiber 

type shift, i.e. towards a slower phenotype, as in ageing 

limb muscles occurs also in an ageing SCM (Figure 

36),244 despite a significant inactivity atrophy is avoided. 

In the ageing, SCM the percentage of slow-twitch fibres 

was nearly equal to the share of fast-twitch fibres (44.6 

vs. 45.8% for numerical proportion and 57.2 vs. 50.6% 

for area proportion); in comparison, the SCM of young 

males was a fast-twitch muscle, of which approximately 

one-third was slow-twitch fibres and two-thirds were 

fast-twitch fibres (31.5 vs. 60.8% for numerical 

proportion and 38.4 vs. 63.5 % for areal proportion). 

Regarding the subtypes of fast fibres, the numerical 

proportion of hybrid 2a-2x was statistically significantly 

diminished in ageing SCM (14.1 vs. 26.8 %), due to 

decreased expression of MyHC-2x (19 vs. 34.5 % for 

area proportion). The remaining 10% of muscle fibres 

 
 

Fig 35.Pendulum test EMG recordings from a female 

stroke subject. First line before tSCS treatment, 

second line immediately after tSCS treatment and 

third line four to five hours after treatment. 

Second line shows lighter knee angel oscillations 

and the EMG coordinated responses of 

quadriceps and hamstring muscle after tSCS 

therapy. The effect lasts less than five hours. 

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were either 1/2a fibres, fibres co-expressing MyHC-neo 

with adult myosin heavy chain isoforms or 

untraceable/unclassified fibres and were present in 

similar percentages in the ageing and young male SCM. 

A trend towards a smaller fiber diameter was noticed in 

all fiber types in the ageing SCM, but the differences 

were not statistically significant. A slower muscle 

phenotype with the preferential loss of the fibres co-

expressing the fastest myosin isoform MyHC-2x in the 

ageing SCM 6 provide a circumstantial evidence for the 

selective loss of fast-twitch motor neurons in the cervical 

spinal cord and the reinnervation process in the ageing 

SCM. The ageing SCM appears to be an excellent subject 

for studying the effects of ageing on MyHC composition 

without significant inactivity atrophy. 

239. Tomlinson BE, Irving D. The numbers of limb motor 

neurons in the human lumbosacral cord throughout life. 

J Neurol Sci 1977;34:213–9. 

240. Mittal KR, Logmani FH. Age-related reduction in 8th 

cervical ventral nerve root myelinated fiber diameters 

and numbers in man. J Gerontol 1987;42:8–10. 

241. Lexell J, Taylor CC. Variability in muscle fibre areas in 

whole human quadriceps muscle: effects of increasing 

age. J Anat 1991;174:239–49. 

242. Campbell MJ, McComas AJ, Petito F. Physiological 

changes in ageing muscles. J Neurol Neurosurg 

Psychiatry 1973;36:174–82. 

243. D'Antona G, Pellegrino MA, Adami R, et al. The effect of 

ageing and immobilization on structure and function of 

human skeletal muscle fibres. J Physiol 2003;552:499–

511. 

244. Meznaric M, Erzen I, Karen P, Cvetko E. Effect of ageing 

on the myosin heavy chain composition of the human 

sternocleidomastoid muscle. Ann Anat 2018;216:95–9. 

***** 

Fiber typing and isomyosin analyses of rat 

sternomastoideus muscle: Constrains to muscle 

sampling from its peculiar distribution of muscle 

fiber types 

Barbara Ravara1,2, Ugo Carraro 1,2 

1. A&C M-C Foundation, Padova, Italy; 2. CIR Myo, 

Myology Center, University of Padova, Italy  

Email: barbara.ravara@unipd.it 

Key words: neck muscles, rat, histochemistry, isomyosins, 

sternomastoideus 

The sternomastoid muscle (SM) in rodents is known to 

have a peculiar distribution of fiber types with a steep 

gradient from surface to deep region.245 We better 

characterized this peculiar regional distribution by 

quantitative histochemical analyses (Figure 37). 

Transverse sections of the SM muscle stained by SDH 

reaction clearly show two distinct regions, toward the 

deep surface of the muscle a 40% area that contains 

packed SDH-positive myofibers, while the other 60% 

area of the SM toward the external surface presents with 

a more checker-board appearance. In the deep region of 

SM the type 1 (slow type) muscle fibers identified by 

positive acidic ATPase pH 4.35 are only the 24.5% of the 

fiber in the deep area of SM muscle and they are 

 

 
 

Fig 36. Numerical proportion (%) of slow- and fast-twitch fibres and their subtypes based on the expression of 

myosin heavy chains.  

mailto:barbara.ravara@unipd.it


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65 

 

restricted in the deepest region. The 75.5% of the 

myofibers in the deep region are of the fast contracting 

types (48.4% 2A, SDH –positive fibers and 27.1% 2B, 

SDH-negative fibers, respectively). Based on present and 

previous observations changes in absolute and relative 

number of fiber types, in any experimental model,246 will 

ask for morphometry of the whole muscle cross-section, 

while after muscle sampling only the size of the different 

types of muscle fibers could be computed.247 

245. Polican Ciena A, Yokomito de Almeida SR, de Matos 

Alves PH, et al. Histological and ultrastructural changes 

of sternomastoid muscle in aged wistar rats. Micron 

2011;42:871-6.. 

246.  Kern H, Loefler S, Hofer C, et al. FES Training in Aging: 

interim results show statistically significant 

improvements in mobility and muscle fiber size. Eur J 

Transl Myology 2012;22:61-7. 

247. Mosole S, Carraro U, Kern H, et al 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. Eur J Transl Myol 2016;26:5972. doi: 

10.4081/ejtm.2016.5972. eCollection 2016 Sep 15. 

Fighting muscle weakness in premature and 

advanced aging by take-home strategies: Full-body 

In-Bed Gym 

Ugo Carraro (a,b), Karma Gava (c) 

(a) IRCCS Fondazione San Camillo Hospital, Venice, 

Italy; (b) CIR-Myo University of Padova, Italy; A&C 

M-C Foundation for Translational Myology, Padova; 

(d) Documentary filmaker, Padova, Italy 

Email: ugo.carraro@unipd.it  

Key Words: Fighting muscle weakness, premature and 

advanced aging, take-home strategies, Full-

body In-Bed Gym 

All progressive muscle contractile impairments need 

permanent managements, including aging-related 

muscle-strength decline. Beside the eventual 

pharmacology therapy, a home-based physical exercise 

approach is helpful and education of hospitalized patients 

to take-home physical exercise managements is an 

effective low cost alternative. Frail elderly persons due to 

advanced age or associated diseases are often 

hospitalized for long periods of time. There, their already 

modest amount of daily physical activity is reduced, 

contributing to limit their independence up to force them 

to bed. Immobility is associated with neuromuscular 

weakness, functional limitations, thromboembolism and 

high costs.248-250 Inspired by the proven capability to 

recover skeletal muscle strength by home-based 

Functional Electrical Stimulation even in the worse cases 

of neuromuscular traumatic injuries,251 we suggest a brief 

(15-20 minutes) daily routine of easy-to-perform 

 
 
Fig 38. Active persons, able to make 20 consecutive push-

up in 3 minutes (Figure 1, V), do not need In-Bed-

Gym, but sedentary people may gradually start with 

5 repetition, and add weekly  up to 20.  

 
 

 
 

Fig 37. Transverse cross-section of normal rat 

Sternomastoid muscle 

 Upper panel: SDH reaction 

 Lower panel: ATPase pH 4.35. 

mailto:ugo.carraro@unipd.it


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66 

 

physical exercises that may be performed in bed: Full-

body In-Bed Gym.252,253 Full-body In-bed Gym is an 

extension to all body muscles of well-established 

physiotherapy approaches of in-bed cardio-circulation-

ventilation workouts (Figure 38). If sedentary borderline 

persons challenge them-self, in hospital Full-body In-

Bed Gym may increase muscle strength and fatigue 

resistance. In surgical units this will grant standing of 

patients soon after operation, a mandatory measure to 

prevent risk of thromboembolism. Full-body In-Bed 

Gym helps also to mitigate the bad mood that 

accompanies mobility limitations, strengthening 

patients’ confidence in recovering partial or total 

independence. Continued regularly, Full-body In-Bed 

Gym may help to maintain independence of older people 

reducing the risks of the possible serious consequences 

of accidental falls. 

248. Hopkins RO, Mitchell L, Thomsen GE, et al. 

Implementing a Mobility Program to Minimize Post–

Intensive Care Syndrome. AACN Adv Crit Care 

2016;27:187-203. 

249. Camillo CA, Osadnik CR, van Remoortel H, et al. Effect 

of "add-on" interventions on exercise training in 

individuals with COPD: a systematic review. 

250. Carraro U, Kern H, Gava P, et al. Recovery from muscle 

weakness by exercise and FES: lessons from Masters, 

active or sedentary seniors and SCI patients. Aging Clin 

Exp Res 2016 Sep 3. [Epub ahead of print] Review. 

251. Zampieri S, Mosole S, Löfler S, et al. Physical Exercise 

in Aging: Nine Weeks of Leg Press or Electrical 

Stimulation Training in 70 Years Old Sedentary Elderly 

People. Eur J Transl Myol 2015; 25: 237–242. doi: 

10.4081/ejtm.2015.5374. 

252. Carraro U, Gava K, Baba A, Piccione F, Marcante A. 

Fighting muscle weakness in advanced aging by take-

home strategies: Safe anti-aging full-body in-bed gym 

and functional electrical stimulation (FES) for mobility 

compromised elderly people. Biol Eng Med 2016 1; 1-4. 

doi: 10.15761/BEM.1000106. 

253.  Carraro U, Gava K, Musumeci et al. A Safe Antiaging 

Full-Body In-Bed Gym and FES for Lazy Persons: Home 

In-Bed Exercises for Fighting Muscle Weakness in 

Advanced Age. . In: Masiero S., Carraro U. (eds) 

Rehabilitation Medicine for Elderly Patients. Practical 

Issues in Geriatrics. Springer, Cham 2018; pp 213-221. 

https://doi.org/10.1007/978-3-319-57406-6_6. 

***** 

Altered differentiation of primary myocytes in an 

ALS mouse model 

Caterina Peggion (a), Roberto Stella (b), Kelly Nies (a), 

Maria Catia Sorgato (a,c), Alessandro Bertoli (a,c,d), 

Maria Lina Massimino (c) 

(a) Dept. of Biomedical Sciences, University of Padova, 

Italy; (b) Istituto Zooprofilattico Sperimentale delle 

Venezie, Legnaro (PD), Italy; (c) Neuroscience 

Institute, CNR, Padova, Italy; (d) Padova Neuroscience 

Center, University of Padova, Italy 

Email: alessandro.bertoli@unipd.it  

Key Words: Amyotrophic lateral sclerosis, differentiation, 

primary myocytes, ALS mouse model 

Amyotrophic lateral sclerosis (ALS) is a neuro-

degenerative disease characterized by motor neuron 

(MN) degeneration, muscle atrophy, paralysis and, 

ultimately, death. While the great majority of ALS cases 

are sporadic (sALS), 10% are inherited (fALS), and 

approximately 20% of fALS are caused by mutations in 

the Cu/Zn superoxide dismutase 1 (SOD1) gene. 

Accumulating evidence suggests that non-cell-

autonomous processes involving the interaction with 

neighbouring cells contribute to MN pathology in ALS, 

possibly through “dying-back” mechanisms. In addition, 

numerous studies indicate that skeletal muscle is one of 

the primary targets for mutant SOD1 (mSOD1) toxicity 

both in human sALS and fALS,254,255 and in transgenic 

(Tg) mice.256,257 Little is known about muscle fibre 

regeneration capacity in ALS. Muscle skeletal 

regeneration is an important homeostatic process that 

guarantees maintenance of muscle integrity and 

 
 

Fig 39. After 4 days of differentiation in vitro, 

primary myocytes from neonatal 

hSOD1(G93A) (upper panel) and 

hSOD1(WT) (lower panel) Tg mice, were 

fixed, permeabilised, immuno-stained with 

an antibody to embryonic myosin heavy 

chain (eMyHC) (red signal), and then 

counter-stained with the nuclear fluoro-

probe Hoechst-33342 (blue signal). It is 

evident that cultures from hSOD1(G93A)-

expressing mice contain less eMyHC-

positive myotubes compared to the 

hSOD1(WT) control. Scale bar = 20 μm. 

 

mailto:alessandro.bertoli@unipd.it


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plasticity. In normal muscles, satellite cells became 

activated, proliferate and differentiate in response to 

tissue damage to regenerate muscle fibers. A satellite cell 

impairment, with consequent less efficient regeneration 

of skeletal muscles, however, has been recently 

demonstrated in an ALS mouse model expressing the 

SOD1(G93A) mutant.258 Such a reduced regenerative 

potential could limit the efficacy of compensatory 

processes, thereby contributing to the progression and/or 

severity of muscle atrophy and weakness. To determine 

if mSOD1 perturbs the myogenic program, we carried 

out an in vitro study using primary cultured myocytes 

from neonatal Tg mice expressing human (h) 

SOD1(G93A) or the wildtype (WT) hSOD1 counterpart. 

We found that, 4 days after switching to a differentiative 

culture medium, hSOD1(G93A) cells show less 

differentiated myotubes, and reduced expression of 

embryonal myosin heavy chain, compared to 

hSOD1(WT) cultures (Figure 39). The finding that 

expression of both Pax7 and MyoD is decreased in 2 day-

old cultured mSOD1 myocytes, and preliminary results 

from proliferation assays suggest that SOD1(G93A) 

myoblasts suffer from defective proliferation in the early 

stages of the myogenic program. As a future perspective, 

we envisage to get insight into the molecular mechanism 

that may perturb adult myogenesis in SOD1(G93A) mice 

by analysing miRNAs (and targets thereof) that are 

known both to play a leading role in the myogenic 

program, and to be altered in ALS (i.e., miR1, miR206, 

miR133a).259 

254. Corti S, Donadoni C, Ronchi D, et al. Amyotrophic lateral 

sclerosis linked to a novel SOD1 mutation with muscle 

mitochondrial dysfunction. J Neurol Sci 2009;276:170–4. 

255. Vielhaber S, Kornblum C, Heinze HJ, et al. Mitochondrial 

changes in skeletal muscle in amyotrophic lateral sclerosis 

and other neurogenic atrophies: a comment. Brain 

2005;128:E38. 

256. Dobrowolny G, Aucello M, Rizzuto E, et al. Skeletal 

muscle is a primary target of SOD1G93A-mediated toxicity. 

Cell Metab 2008;8:425–36. 

257. Wong M, Martin LJ. Skeletal muscle-restricted 

expression of human SOD1 causemotor neuron 

degeneration in transgenic mice. Hum Mol Genet 

2010;19:2284–302. 

258. Manzano R, Toivonen JM, Calvo AC, et al. () Altered in 

vitro proliferation of mouse SOD1-G93A skeletal muscle 

satellite cells. Neurodegener Dis 2013;11:153–64. 

259. Martini M, Dobrowolny G, Aucello M, Musarò A 

Postmitotic Expression of SOD1(G93A) Gene Affects the 

Identity of Myogenic Cells and Inhibits Myoblasts 

Differentiation. Mediators Inflamm 2015;2015:537853. 

***** 

MicroRNA expression in genetic and sporadic ALS 

Laura Giaretta, Valentina Pegoraro, Corrado Angelini 

San Camillo Hospital IRCCS, Venice, Italy 

Email: corrado.angelini@ospedalesancamillo.net  

Key Words: MicroRNA expression, genetic and sporadic ALS 

Amyotrophic Lateral Sclerosis (ALS) is a progressive 

neurodegenerative disease characterized by the 

degeneration of upper and lower motor neurons and the 

progressive loss of synaptic connection between nerve 

and muscle. The disease leads to a decline in strength, 

severe muscle atrophy, paralysis and death within 3-5 

years after its diagnosis. While the majority of ALS cases 

are sporadic (SALS), about 10% of ALS cases have a 

familial inheritance (FALS). Among the familial cases 

the most frequent genetic cause is associated with an 

abnormal repeat in the 3’ untranslated region of C9orf72 

gene. The role of this expansion in ALS disease is still 

unclear: it is possible that the expansion causes a 

reduction of C9orf72 protein that in turn contributes to 

the disease; another more convincing hypothesis is that 

the abnormal repeats causes the accumulation of 

intranuclear RNA-foci sequestering RNA-binding 

proteins as a consequence; a third possible mechanism 

proposes that the expansion causes the accumulation of 

toxic dipeptide repeat (DPR) protein.260,261 Another 

frequent genetic cause leading to ALS is due to mutation 

in the gene SOD1, coding for a superoxide dismutase 

enzyme with an important antioxidant function. 

Although the exact mechanism that leads to motor 

neurons degeneration is unknown, the general consensus 

is that the toxicity arises from the accumulation of the 

mutant SOD1 protein leading to motor neurons 

degeneration.262 MicroRNAs are small non-coding 

RNAs that regulate the expression of specific genes by 

binding to the 3’ untranslated region of the target mRNA 

(Figure 40). The binding leads to the cleavage of the 

target mRNA or to translation impairment, therefore an 

increase of a specific microRNA leads to the decrease of 

the corresponding mRNA and to the down-regulation of 

the corresponding protein. Some microRNAs are 

expressed in a tissue-specific manner. In muscle a group 

of microRNAs called myomiRNA regulate specific 

myogenic processes such as skeletal muscle 

proliferation, myogenic differentiation and atrophy.263 

 
 
Fig 40. MicroRNA in ALS muscle are diversely 

expressed in SALS and FALS patients, with 

a significant increase in muscle-specific 

miRNAs, miR-206 and miR-27a, and 

inflammatory miRNAs, miR-155 and miR-

221 in FALS samples. 

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2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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68 

 

Other microRNAs, such as miR-155, miR-146a and miR-

221, are involved in the regulation of the immune system 

and in the inflammatory response during muscle damage 

or disease promoting the regeneration process.264,265 In 

this study we analyzed the expression of five muscle-

specific miRNA (miR-1, miR-206, miR133a, miR-133b, 

miR-27a) and of three inflammatory miRNA (miR-155, 

miR-146a and miR-221) in muscle biopsies of 18 ALS 

patients with different genetic form of ALS disease: four 

carried the C9orf72 hexanucleotide repeat expansion 

(C9-ALS), four carried the SOD1 mutation (SOD1-ALS) 

and other 10 were considered SALS. Our results show 

different expression of miR-206 and miR-27a in genetic 

(C9-ALS and SOD1-ALS) against SALS. Inflammatory 

miRNAs were also up-regulated in FALS cases, in 

particular there was a significant increase in miR-155 and 

miR-221 in SOD1-ALS against SALS and miR-221 in 

C9-ALS. Our data suggest that a diverse regulatory 

mechanism may exist between different forms of ALS. 

260.  Taylor JP, Brown RH, Cleveland DW. Decoding ALS: 

from genes to mechanism. Nature 2016;539:197–206, 

doi:10.1038/nature20413. 

261.  DeJesus-Hernandez M, Mackenzie IR, Boeve BF,et al. 

Expanded GGGGCC Hexanucleotide Repeat in 

Noncoding Region of C9ORF72 Causes Chromosome 9p-

Linked FTD and ALS. Neuron 2011;72:245–56, 

doi:10.1016/j.neuron.2011.09.011. 

262.  Bruijn LI, Houseweart MK, Kato S,et al. Aggregation and 

motor neuron toxicity of an ALS-linked SOD1 mutant 

independent from wild-type SOD1. Science 

1998;281:1851–4. 

263.  Horak M, Novak J, Bienertova-vasku J. Muscle-specific 

microRNAs in skeletal muscle development. Dev Biol 

2016;410:1–13, doi:10.1016/j.ydbio.2015.12.013. 

264.  Nie M, Liu J, Yang Q, et al. MicroRNA-155 facilitates 

skeletal muscle regeneration by balancing pro- and anti-

inflammatory macrophages. Cell Death Dis 

2016;7:e2261, doi:10.1038/cddis.2016.165. 

265.  Cardinali B, Castellani L, Fasanaro P,et al. Microrna-

221 and microrna-222 modulate differentiation and 

maturation of skeletal muscle cells. PLoS One 2009;4, 

doi:10.1371/journal.pone.0007607. 

***** 

Distinct pattern of microRNA expression in Becker 

dystrophy and LGMD 

Roberta Marozzo, Valentina Pegoraro, Laura Giaretta, 

Corrado Angelini 

San Camillo Hospital IRCCS, Venice, Italy 

Email: corrado.angelini@ospedalesancamillo.net 

Key Words: MicroRNA expression, Becker dystrophy, LGMD 

Becker muscular dystrophy (BMD) is an X-linked 

recessive inherited disorder, due to a mutation in 

dystrophin gene that encodes for the sarcolemmal protein 

dystrophin. This mutation leads to translation of a 

truncated dystrophin, which is expressed at lower and 

more variable levels than full-length dystrophin.  

Individuals with this disorder typically present 

progressive weakness and wasting of the skeletal and 

cardiac muscles.266,267 However, some BMD cases have 

no overt muscle weakness and wasting and their 

independent activity is maintained until late adulthood.268 

The limb-girdle muscular dystrophies (LGMDs) are an 

heterogeneous group of rare progressive genetic 

muscular disorders that are characterized by a 

progressive weakness with onset in the proximal limb 

girdle muscles.269 Two subgroups of limb-girdle 

muscular dystrophies (LGMDs) are calpainopathy 

caused by mutations in the CAPN3 gene which encodes 

the skeletal muscle–specific member of the calpain 

family,270 and sarcoglycanopathies in which the primary 

defect is in one sarcoglycan (SG) transmembrane 

glycoprotein (alpha, beta, gamma, delta) and results in a 

deficiency of the whole sarcoglycan complex. We 

studied sarcoglycanopathies due to mutations in genes 

encoding two of these glycoproteins, more precisely the 

beta-sarcoglycan protein and gamma-sarcoglycan 

protein. MicroRNAs (miRNAs) are small non-coding 

RNA molecules approximately 22 nucleotides in length, 

which are involved in gene expression at the post-

transcriptional level and regulate many cellular 

functions. A group of miRNAs are highly expressed in 

skeletal and cardiac muscle and they are called myomiRs. 

The myomiR family includes miR-1, miR-133a, miR-

133b, miR-206 which are used as non-invasive 

biomarkers in neuromuscular diseases.271 We analysed 

miRNA expression level by Real Time PCR in the serum 

of 4 BMD patients (Table 3), 6 LGMD patients (4 female, 

2 male) and 10 control subjects. The most highly 

dysregulated serum miRNA in BMD was miR-206, a 

skeletal muscle–specific miRNA. One patient affected by 

BMD, who presented a major clinical and MRI 

alterations with calf  hypertrophy, showed a marked 

increase of miR-206 and miR-133b and a slight up-

regulation of the other miRNA, compared to the control 

group. In LGMD cases it was possible to observe an 

increase of miR-133a and miR-206 in two patients 

affected respectively by beta-sarcoglycanopathy and 

gamma-sarcoglycanopathy while miR-1 and miR-133b 

levels exhibited no significant dysregulation. These data 

highlight the potential use of miRNA as biomarkers of 

BMD and LGMD. 

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2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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69 

 

266. Barp A, Bello L, Politano L, et al. Genetic Modifiers of 

Duchenne Muscular Dystrophy and Dilated 

Cardiomyopathy. PLoS One 2015;10:e0141240.  

267  Melacini P, Fanin M, Angelini A, et al. Cardiac 

transplantation in a Duchenne muscular dystrophy 

carrier. Neuromuscul Disord. 1998;8:585-90. 

268  Bello L, Campadello P, Barp A, et al. Functional 

changes in Becker muscular dystrophy: implications for 

clinical trials in dystrophinopathies. Sci Rep 

2016;6:32439.  

269  Angelini C, Tasca E, Nascimbeni AC, Fanin M. Muscle 

fatigue, nNOS and muscle fiber atrophy in limb girdle 

muscular dystrophy. Acta Myol 2014;33:119-26. Review  

270 Fanin M, Angelini C. Protein and genetic diagnosis of 

limb girdle muscular dystrophy type 2A: The yield and the 

pitfalls. Muscle Nerve. 2015;52:163-73. 

271  Alexander MS, Kunkel LM. Skeletal Muscle 

MicroRNAs: Their Diagnostic and Therapeutic Potential 

in Human Muscle Diseases. J Neuromuscul Dis 

2015;2:1-11. 

***** 

Rehabilitation in DM1 patients: observation in 

circulating myomiRNAs and myostatin 

Valentina Pegoraro, Laura Giaretta, Paola Cudia, 

Alfonc Baba, Corrado Angelini 

San Camillo Hospital IRCCS, Venice, Italy 

Email:corrado.angelini@ospedalesancamillo.net 

Key Words: Rehabilitation, DM1 patients,: circulating 

myomiRNAs, myostatin 

Myotonic Dystrophy type 1 (DM1), a multi-systemic 

autosomic dominant disorder caused by a trinucleotide 

(CTG) expansion, is the most common inherited 

muscular dystrophy in adulthood. DM1 is characterized 

mainly by progressive muscular weakness, myotonia, 

respiratory failure, cardiac defects, cataracts and cerebral 

involvement. MicroRNAs (miRNAs) are small non-

coding RNAs that regulate post-transcriptional mRNA 

typically by binding to the 3’-untranslated region of the 

complementary mRNA sequence, resulting in 

translational repression and gene silencing. Therefore, an 

increase in a specific miRNA results in a decreased 

expression of the corresponding protein product.  

MiRNAs have recently gained attention for their 

potential as minimally invasive and cost-effective disease 

biomarkers. MiRNAs are known to be secreted by 

various cell types and, unlike most mRNAs, they are 

markedly stable in circulating body fluids due to proteic 

protection from ribonucleases. Because of their stability 

in plasma and serum, they can be reliably detected even 

at low concentration and used not only as markers of 

disease but also to measure the effectiveness of novel 

drug therapies and rehabilitation. MiR-1, miR-206, miR-

133a and miR-133b are called “myomiRNAs” and are 

involved in myogenesis, manteinement and recovery of 

muscles. In DM1 patients miRNA have been extensively 

studied in both muscle and serum/plasma samples 

demonstrating that their expression level correlate with 

loss of muscle strength and with the different disease 

stage.272-274 Perfetti and collaborators found an up-

regulation of myomiRNA in DM1 patients. Myostatin, 

also known growth factor 8 (GDF8), belongs to the TGF-

β (transforming growth  factor-β) superfamily and it acts 

as negative regulator of muscle mass and it could be used 

as an important indicator of muscle atrophy. Recently it 

was reported that myostatin is involved in regulation of 

pathways of skeletal muscle mass induced by exercise 

training.275 MyomiRNAs and myostatin are considered 

possible biomarkers of muscle atrophy.276 Rehabilitative 

intervention is useful to optimize muscle trophism and to 

 

Table 3. Becker muscular dystrophy patients: clinical, radiological and miRNAs data 
 

 

  = Up-regulation 

 Age MRI changes Treatment miRNAs changes 

Patient 1 41 +++ steroids 

 

miR-1 

miR-206 

miR-133a 

miR-133b 

 

Patient 2 39 ++ steroids 
 

miR-206 

Patient 3 32 no changes 

 

no significant 

dysregulation 

Patient 4 41 + 
  

miR-206 

 

 

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2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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70 

 

prevent additional disuse muscle atrophy. Aerobic and 

resistance training as well as Functional electrical 

stimulation (FES) combined with cycling improved 

muscle function and strength in patients with DM1.277 In 

this study we collected serum and plasma of 10 DM1 

patients (9 male, 1 female) before (T0) and after (T1) a 

period of 3-6 weeks of physical FES/aerobic 

rehabilitation. We measured circulating muscle-specific 

microRNAs by qRT-PCR and myostatin by ELISA.  To 

evaluate endurance and gait speed we used the 6-minute 

walking test and the time 10-m walk test respectable. 

Both myomiRNAs (miR-1, miR-206, miR-133a and 

miR-133b) and myostatin significantly decreased after 

aerobic/FES rehabilitation and the DM1 patients showed 

an improvement of muscle strength and fitness 

performance Our study suggest that myomiR and 

myostatin levels could support clinical data. MicroRNAs 

should be considered a good serum biomarkers of 

functional response to rehabilitation. 

272. Perfetti A, Greco S; Cardani R, et al. Erratum: Validation 

of plasma microRNAs as biomarkers for myotonic 

dystrophy type 1. Sci Rep 2017;7:43074, 

doi:10.1038/srep43074. 

273. Fritegotto C, Ferrati C, Pegoraro V, Angelini C. Micro-

RNA expression in muscle and fiber morphometry in 

myotonic dystrophy type 1. Neurol Sci 2017;38:619-25, 

doi:10.1007/s10072-017-2811-2 

274. Gambardella S, Rinaldi F, Lepore SM, et al. 

Overexpression of microRNA-206 in the skeletal muscle 

from myotonic dystrophy type 1 patients. J Transl Med 

2010;8:48. doi: 10.1186/1479-5876-8-48. 

275. Allen DL, Hittel DS, McPherron AC. Expression and 

Function of Myostatin in Obesity, Diabetes and Exercise 

Adaptation. Med sci Sport Exerx 2011;43:1828-35, 

doi:10.4238/2011. 

276. Tasca E, Pegoraro V, Merico, A, Angelini C. Circulating 

microRNAs as biomarkers of muscle differentiation and 

atrophy in ALS. Clin Neuropathol 2016;35:22–30, 

doi:10.5414/NP300889. 

277. Cudia P, Weis L, Baba A, et al. Effects of Functional 

Electrical Stimulation Lower Extremity Training in 

Myotonic Dystrophy Type I: A Pilot Controlled Study. Am 

J Phys Med Rehabil 2016;95:809–17, 

doi:10.1097/PHM.0000000 000000497. 

***** 

Role of nutrition in neurodegenerative diseases 

Vittorio Emanuele Bianchi (a) , Mauro Tercon (b), 

Laura Rizzi (c), Matteo Toneatto (d), Stefano Inglese (e) 

(a) Endocrinology and Metabolism, Clinical Center 

Stella Maris, Falciano, San Marino; (b) Medical Center 

“Riabilita”, Forlimpopoli (FO); (c) Molecular Biology, 

School of Medicine and Surgery, University of Milano-

Bicocca, Monza Brianza; (d) Academy Manual 

Therapies, Busto Arsizio (MI); (e) Fisiogym Clinik, 

Rehabilitation Center, Metauro (PU), Italy. 

E-mail: dott.vbianchi@gmail.com 

Neurodegenerative diseases are characterized by the 

progressive loss of neuronal function in the brain causing 

cognitive impairment. The most common form is 

Alzheimer disease but is also included amyotrophic 

lateral sclerosis (ALS) and Parkinson disease (PD). 

Despite inflammation and hormonal deficiencies play an 

essential role in the pathogenesis of neurodegeneration, 

nutrition is one of the modifiable factors that has been 

included in the physiopathology of these diseases. A 

systematic literature search was performed using 

PubMed Medline and Cochrane Central Register of 

Controlled Trials. A combination of the following 

keywords was used: “nutrition” with “Alzheimer’s’ 

disease,” “nutrition” with Amyotrophic lateral sclerosis,” 

and “nutrition” with “neurodegeneration.” The search 

included the filter “clinical trials” and “humans.”  

Uncompleted studies that did not evaluate the mental 

impairment have been excluded. We found 124 articles, 

of these 27 have been selected including 9058 patients 

with a mean age of 71,5±7,7. A significant difference 

between the duration of the studies, (varying from 3 

weeks until 51 months) and the number of patients (from 

24 until 2911) has been observed. Furthermore, in 

various studies, information about the type of diet, 

macronutrients, and calories ingested by the patients are 

lacking.  The most common date emerged is that 

malnutrition and low body mass index correlated with the 

higher development of dementia and mortality, showing 

that nutrition is involved in the neurodegenerative 

process and SLA.278 The administration of 

 
 

Fig 41. Experimental flow-chart of the rehabilitation protocol: the DM1 patients make aerobic/FES rehabilitation: 

before and after training blood sample were collected to analyze myomiRNA and myostatin. Both 

myomiRNAs (miR-1, miR-206, miR-133a and miR-133b) and myostatin significantly decreased after 

aerobic/FES rehabilitation 

mailto:dott.vbianchi@gmail.com


2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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71 

 

polyunsaturated fatty acids, either alone or in 

combination, had no significant effects on cognitive 

decline. High-dose B vitamin supplementation seemed 

beneficial in patients with cognitive dysfunction. Also, 

the administration of ginkgo biloba did not support any 

improving cognitive function A high-glycemic diet was 

associated with the higher cerebral amyloid burden and 

AD development. Dietary supplementation with protein 

showed a positive effect on cognitive function. The 

pathogenesis of motoneuron degeneration is not yet 

entirely understood, but nutrition represents a critical co-

factor regulating the development of the disease. High 

protein diet and ketogenic diet seem to be the most 

effective increasing the IGF-1 plasma level and 

stimulating the IGF-1 receptor expression in the brain.279 

Circulating IGF-1 level has a protective effect on the 

brain and mediates the formation of new neurons in the 

adult hippocampus.280 Insulin signaling is a specific 

independent inhibitor of neurons regeneration in aging,281 

while the insulin/IGF-1 signaling has a neuroprotective 

effect.282 So that favoring lower insulin and higher IGF-1 

plasma level has a protective effect on the neuron, and a 

low carbohydrates-high protein diet seems to support this 

hormonal attitude while a high-fat diet predisposes to 

neuroinflammation in central and peripheral nervous 

systems. Nutrition plays a fundamental role in 

maintaining brain health and reduce neurodegenerative 

diseases development. However, an insufficient number 

of the clinical trial has investigated the interaction of 

macronutrients, hormonal impact and the decline of brain 

function. The drawback is represented by the small group 

of patients evaluated, too short time of observation and 

limited protocols with insufficient information about 

macronutrients ingested hormonal evaluation and 

cognitive evaluation. 

278. Shimizu T, Nagaoka U, Nakayama Y, et al. Reduction rate 

of body mass index predicts prognosis for survival in 

amyotrophic lateral sclerosis: a multicenter study in 

Japan. Amyotroph Lateral Scler 2012;13:363-6. 

279. Calikoglu A, Karayal A, .D'Ercole A. Nutritional 

regulation of IGF-I expression during brain development 

in mice. Pediatr Res 2001;49:197-202. 

280. Trejo JL, Carro E, Torres-Aleman I. Circulating insulin-

like growth factor I mediates exercise-induced increases 

in the number of new neurons in the adult hippocampus. 

J Neurosci 2001;21:1628-34. 

281. Byrne AB, Walradt T, Gardner KE, et al. Insulin/IGF1 

signaling inhibits age-dependent axon regeneration. 

Neuron 2014;81:561-73. 

282. Mishra N, Lata S1, Deshmukh P, et al. Insulin signaling 

pathway protects neuronal cell lines by Sirt3 mediated 

IRS2 activation. Biofactors 2018 Feb 7. doi: 

10.1002/biof.1413. [Epub ahead of print] 

***** 

Regional regulation of mechano-sensitive proteins is 

related to human skeletal muscle remodelling in 

response to concentric vs. eccentric loading 

Martino V Franchi (1), Severin Ruoss  (1), Jonathan I 

Quinlan (2), Martin Flück (1)*, Marco V Narici (3)* 

 (1) Laboratory for Muscle Plasticity, Balgrist 

University Hospital, University of Zurich, Switzerland 

(2) MRC-ARUK Centre of Musculoskeletal Ageing 

Research, University of Nottingham, Derby, UK 

(3) Institute of Physiology, Department of Biomedical 

Sciences, University of Padua, Italy 

*= co-senior authors 

E-mail: martino.franchi@balgrist.ch 

Keywords: Mechanotransduction, Muscle Architecture, 

Focal Adhesion Kinase 

Concentric (CON) vs. eccentric (ECC) resistance 

training (RT) can lead to similar hypertrophy but with 

distinct changes in muscle architecture (i.e., fascicle 

length and pennation angle).283 To gain insights into the 

molecular mechanisms underlying these remodelling 

patterns, we collected biopsies at mid-belly and 4 am 

above distal myotendinous junction (MTJ) of the vastus 

lateralis (VL) muscle at 0, 4 and 8 weeks of either CON 

or ECC RT.284,285 We specifically targeted focal adhesion 

kinase (FAK) and Vinculin isoforms (gamma- and meta-

), mechanosensitive costameric proteins, involved in the 

modulation of muscle remodelling and protein 

synthesis.286,287, The different adaptations to CON and 

ECC RT were reflected by a higher y397-FAK/FAK 

activation and meta-vinculin at the MTJ compared to 

mid-belly.285 Furthermore, changes muscle architecture 

were positively correlated to y397FAK activity only after 

ECC RT and only at the MTJ site.3 In the present study, 

we investigated, using Extended Field of View (EFOV) 

ultrasonography the regional changes in VL architecture 

after 8-weeks of ECC-RT or CON-RT at 60% one 

repetition maximum (1RM ECC and 1 RM CON). 

Sixteen males were randomized into 3 groups: ECC 

(n=6), CON (n=6), and non-training controls (CTRL). 

Longitudinal EFOV images were collected at baseline 

and at 8-wks to assess fascicle length (Lf) and pennation 

angle (PA) at 0-20%, 20-40%, and 40-60% of muscle 

length (Lm), 0 representing the distal MTJ. Changes in 

Lf were heterogeneous along the VL in the ECC RT 

group: 9.6±1.9% at 0-20% Lm (P<0.001), 6±1% at 20-

40% Lm (P<0.05) 3.9±1% at 40-60% Lm (P<0.01), while 

PA showed little change. No changes were observed for 

the CON group. Instead, PA specifically increased after 

CON training and showed distinct regional variations: 

9.4±1.4% at 0-20% Lm (P<0.05), 5±0.6% at 20-40% Lm 

(P<0.05) and 4.3±1% at 40-60% Lm (N.S.). The present 

data provide evidence that the architectural remodelling 

strategies of human muscle to ECC and CON loading are 

region-specific. Together with evidence of activation of 

region and contraction-specific mechanosensitive 

pathways, such data may suggest that pY397FAK and 

meta-vinculin may play a role in orchestrating the pattern 

of muscle remodelling. 

mailto:martino.franchi@balgrist.ch


2018SpPMD: Giovanni Salviati Memorial, March 15-17 

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72 

 

283. Franchi MV, Atherton PJ, Reeves ND, et al. 

Architectural, functional and molecular responses to 

concentric and eccentric loading in human skeletal 

muscle. Acta Physiol (Oxf) 2014;210:642-54. doi: 

10.1111/ apha.12225. 

284. Franchi MV, Wilkinson DJ, Quinlan JI, et al. Early 

structural remodeling and deuterium oxide-derived 

protein metabolic responses to eccentric and concentric 

loading in human skeletal muscle. Phys Reports 

2015;3(11). pii: e12593. doi: 10.14814/phy2.12593. 

285. Franchi MV, et al. Acta Physiol 2018 (in press) 

286. Klossner S, Durieux AC, Freyssenet D, Flueck M. Eur J 

Appl Physiol. 2009;106:389-98. doi: 10.1007/s00421-

009-1032-7. Epub 2009 Mar 18. 

287. Li J, Wilson N, Zelazny A, et al. Assessment of diffraction-

enhanced synchrotron imaging for cartilage 

degeneration of the human knee joint. Clin Anat 

2013;26:621-9. doi: 10.1002/ca.22106. Epub 2012 Jun 4. 

***** 

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Received for publication: February 20, 2018 

Accepted for publication: February 21, 2018.