In the field of ergonomics, biomechanics, sports and rehabilitation muscle fatigue is regarded as an important aspect since muscle fatigue is considered to be one of the main reasons for musculoskeletal disorders. Classical signal processing techniques used to understand muscle behavior are mainly based on spectral based parameters estimation, and mostly applied during static contraction and the signal must be stationary within the analysis window; otherwise, the resulting spectrum will make little physical sense. Furthermore, the shape and size of the analysis window also directly affect the spectral estimation. But fatigue analysis in dynamic conditions is of utmost requirement because of its daily life applicability. It is really difficult to consistently find the muscle fatigue during dynamic contraction due to the inherent non-stationary nature and associated noise in the signal along with complex physiological changes in muscles. Nowadays, in addition to linear signal processing, different non-linear signal processing techniques are adopted to find out the consistent and robust indicator for muscle fatigue under dynamic condition considering the high degree of non-linearity (caused by functional interference between different muscles, changes of signal sources and paths to recording electrodes, variable electrode interface etc.) in the signal. In this work, various linear and nonlinear-non-stationary signal processing methods, applied on surface EMG signal for muscular fatigue analysis under dynamic contraction are studied. In present study, surface EMG (sEMG) signals are recorded from Biceps Brachii muscles from eight (N=8) physically active college students during dynamic lifting 7 kg load at the rate of 20 lifts/min till they become fatigue. EMG data is processed in two ways -1. taking the whole EMG response and 2. breaking into three ranges of contraction (0-45)o, (45-90)o and >90o, to study better response region. It is observed that in spectral estimation techniques auto-regressive (AR) based spectral estimation technique gives better frequency resolution than periodogram for small epochs, as AR is based on parametric estimation. Both the previous methods provide only the frequency information in the signal. In order to estimate the time varying nature of frequency content in a signal various time-frequency signal processing techniques are used like – Short Time-Fourier Transform (STFT), Smoothed pseudo Wigner-Ville (SPWD), Choi-William distribution (CWD), Continuous Wavelet Transform (CWT), Huang-Hilbert Transform (HHT) and Recurrence Quantification Analysis (RQA) are used. The last two techniques are used by considering the EMG signal as non-linear and non-stationary signals. Among these techniques, STFT is the simplest time-frequency analysis technique. But tradeoff between time and frequency resolution is the major constraint in STFT, therefore, a window length of 256 samples are considered in this study. In order to tackle time-frequency resolution problem different Cohen-class distribution techniques are used like SPWD and CWD, where the result is severely affected by the presence of interference terms which make its interpretation really difficult. Different adaptive filters are used in SPWD and CWD to suppress these interference terms during analysis. Among these time-frequency analysis techniques continuous wavelet transform provides the most accurate results in comparison to other time-frequency analysis techniques. Similar result is obtained in present study. This fatigue response is further improved using non-linear and non-stationary techniques like HHT and RQA. HHT shows less variation in frequency response than CWT analysis result. Percentage of determinism calculated using recurrence quantification analysis method is found to be more sensitive than mean frequency estimation. Therefore, non-linear and non-stationary signal processing techniques are to be better indicator of muscle fatigue during dynamic contraction.

Peptides play pivotal roles in the regulation of various physiological processes. As neuropeptides or peptide hormones, they can bind to a range of receptors and thereby trigger the activation of different pathways, including insulin signaling. Another central functionality is facilitated by the action of the as regulins summarized transmembrane micropeptides. By binding to the sarco/endoplasmic reticulum Ca2+ ATPase (SERCA), the regulins control Ca2+ homeostasis and muscle contraction. With the ongoing identification of novel modulatory micropeptides encoded by small open reading frames, the urgency to understand peptide-dependent regulatory networks rises. In this regard, especially impact and physiological relevance exerted by the enzymatic inactivation of the mature, biologically active peptides are far from completely understood. Neprilysins are metalloendopeptidases expressed throughout the animal kingdom. Based on their broad substrate specificity, the activity of neprilysins is crucial for the modulation of multiple peptide-dependent processes. This work aimed to identify new peptide substrates of the Drosophila melanogaster Neprilysin 4 (Nep4) and investigate the enzyme's physiological impact on the affected regulatory mechanisms. The first part of the work could identify 16 novel Nep4 peptide substrates that play essential roles in insulin signaling and the regulation of food intake: allatostatin A1-A4, adipokinetic hormone, corazonin, diuretic hormone 31, drosulfakinin 1 and 2, leucokinin, two short neuropeptide F peptides, and tachykinin 1-4. Thereby, aberrant expression of Nep4 leads to severe phenotypes linked to misregulation of insulin signaling, including reduced body size and weight, compromised food intake, and a characteristic shift in metabolomic composition. To further investigate and understand the complex functionality of the newly discovered Nep4 substrates, these peptides were tested for their ability to modulate the Drosophila heartbeat. A combined in vitro/in vivo screen revealed that the tested substrates exert chronotropic as well as inotropic effects, rendering the peptides as essential novel modulators of the heartbeat in Drosophila. The main project of this thesis was based on the initial finding that animals with Nep4 overexpression exhibit severe impairments of body wall muscle and heart functionality. By applying various experiments, including analyses of muscle and heart contraction, measurement of Ca2+ transients, pull-down studies, STED super-resolution microscopy, and mass spectrometry, Neprilysin 4 was identified as a novel modulator of SERCA activity. The molecular underpinning of this regulatory mechanism is the Nep4 mediated cleavage and inactivation of Drosophila SERCA-inhibitory Sarcolamban micropeptides SCLA and SCLB. Strikingly, cleavage experiments using the mammalian neprilysin and apparent colocalization of Neprilysin and SERCA in human heart tissue indicate evolutionary conservation of this mechanism. In summary, this work could identify a range of so far unknown Nep4 substrates and thereby point out the critical roles these class of enzymes plays in insulin signaling as well as the physiology of muscle and heart contraction.

<p>Asthma is a disease characterized by transient airway smooth muscle contraction leading to episodes of reversible airway narrowing. It affects over 300 million people worldwide and is implicated in over 250 000 deaths annually. The primary clinical features of asthma include airway inflammation, hyperresponsiveness, and remodeling. Generally, asthmatic patients experience exacerbations between periods of diminished symptoms. Interestingly, in addition to these above mentioned hallmarks, asthmatics have also been shown to react differently to ventilatory mechanical strain. This is most evident when assessing the effect of a deep inspiration (DI), clinically measured as a breath taken from functional residual capacity to total lung capacity, in healthy individuals <em>versus</em> asthmatics. These deep inspiratory efforts have been shown to produce a bronchodilatory response in healthy individuals, whereas in asthmatics, DIs are less effective in producing bronchodilation, can cause more rapid airway re-narrowing, and even bronchoconstriction in moderate to severe asthmatics. The mechanism by which a DI is able to cause bronchoconstriction remains ambiguous. Previous theories suggest that this phenomenon is intrinsic to airway smooth muscle (ASM) itself. However, the airway inflammation present in asthmatic airways may also add to the increased ASM contractility following stretch, by the release of mediators that can prime the contractile apparatus to react excessively in the presence of stretch.</p> <p>Thus, collectively, the studies contained in this thesis are linked to the general theme of greater characterization of the signalling mechanisms that regulate airway stretch-activated contractions using a pharmacological approach in intact bovine bronchial segments, with the hope of providing novel insights into the mechanisms that regulate the DI-induced bronchoconstriction seen in asthmatics.</p> / Doctor of Philosophy (Medical Science)

Les systèmes neuromusculaire et musculosquelettique sont des systèmes de systèmes complexes qui interagissent parfaitement entre eux afin de produire le mouvement. En y regardant de plus près, ce mouvement est la résultante d'une force musculaire créée à partir d'une activation du muscle par le système nerveux central. En parallèle de cette activité mécanique, le muscle produit aussi une activité électrique elle aussi contrôlée par la même activation. Cette activité électrique peut être mesurée à la surface de la peau à l'aide d'électrode, ce signal enregistré par l'électrode se nomme le signal Électromyogramme de surface (sEMG). Comprendre comment ces résultats de l'activation du muscle sont générés est primordial en biomécanique ou pour des applications cliniques. Évaluer et quantifier ces interactions intervenant durant la contraction musculaire est difficile et complexe à étudier dans des conditions expérimentales. Par conséquent, il est nécessaire de développer un moyen pour pouvoir décrire et estimer ces interactions. Dans la littérature de la bioingénierie, plusieurs modèles de génération de signaux sEMG et de force ont été publiés. Ces modèles sont principalement utilisés pour décrire une partie des résultats de la contraction musculaire. Ces modèles souffrent de plusieurs limites telles que le manque de réalisme physiologique, la personnalisation des paramètres, ou la représentativité lorsqu'un muscle complet est considéré. Dans ce travail de thèse, nous nous proposons de développer un modèle biofidèle, personnalisable et rapide décrivant l'activité électrique et mécanique du muscle en contraction isométrique. Pour se faire, nous proposons d'abord un modèle décrivant l'activité électrique du muscle à la surface de la peau. Cette activité électrique sera commandé par une commande volontaire venant du système nerveux périphérique, qui va activer les fibres musculaires qui vont alors dépolariser leur membrane. Cette dépolarisation sera alors filtrée par le volume conducteur afin d'obtenir l'activité électrique à la surface de la peau. Une fois cette activité obtenue, le système d'enregistrement décrivant une grille d'électrode à haute densité (HD-sEMG) est modélisée à la surface de la peau afin d'obtenir les signaux sEMG à partir d'une intégration surfacique sous le domaine de l'électrode. Dans ce modèle de génération de l'activité électrique, le membre est considéré cylindrique et multi couches avec la considération des tissus musculaire, adipeux et la peau. Par la suite, nous proposons un modèle mécanique du muscle décrit à l'échelle de l'Unité Motrice (UM). L'ensemble des résultats mécaniques de la contraction musculaire (force, raideur et déformation) sont déterminées à partir de la même commande excitatrice du système nerveux périphérique. Ce modèle est basé sur le modèle de coulissement des filaments d'actine-myosine proposé par Huxley que l'on modélise à l'échelle UM en utilisant la théorie des moments utilisée par Zahalak. Ce modèle mécanique est validé avec un profil de force enregistré sur un sujet paraplégique avec un implant de stimulation neurale. Finalement, nous proposons aussi trois applications des modèles proposés afin d'illustrer leurs fiabilités ainsi que leurs utilité. Tout d'abord une analyse de sensibilité globale des paramètres de la grille HDsEMG est présentée. Puis, nous présenterons un travail fait en collaboration avec une autre doctorante une nouvelle étude plus précise sur la modélisation de la relation HDsEMG/force en personnalisant les paramètres afin de mimer au mieux le comportement du Biceps Brachii. Pour conclure, nous proposons un dernier modèle quasi­ dynamique décrivant l'activité électro-mécanique du muscle en contraction isométrique. Ce modèle déformable va actualiser l'anatomie cylindrique du membre sous une hypothèse isovolumique du muscle. / The neuromuscular and musculoskeletal systems are complex System of Systems (SoS) that perfectly interact to provide motion. From this interaction, muscular force is generated from the muscle activation commanded by the Central Nervous System (CNS) that pilots joint motion. In parallel an electrical activity of the muscle is generated driven by the same command of the CNS. This electrical activity can be measured at the skin surface using electrodes, namely the surface electromyogram (sEMG). The knowledge of how these muscle out comes are generated is highly important in biomechanical and clinical applications. Evaluating and quantifying the interactions arising during the muscle activation are hard and complex to investigate in experimental conditions. Therefore, it is necessary to develop a way to describe and estimate it. In the bioengineering literature, several models of the sEMG and the force generation are provided. They are principally used to describe subparts of themuscular outcomes. These models suffer from several important limitations such lacks of physiological realism, personalization, and representability when a complete muscle is considered. In this work, we propose to construct bioreliable, personalized and fast models describing electrical and mechanical activities of the muscle during contraction. For this purpose, we first propose a model describing the electrical activity at the skin surface of the muscle where this electrical activity is determined from a voluntary command of the Peripheral Nervous System (PNS), activating the muscle fibers that generate a depolarization of their membrane that is filtered by the limbvolume. Once this electrical activity is computed, the recording system, i.e. the High Density sEMG (HD-sEMG) grid is define over the skin where the sEMG signal is determined as a numerical integration of the electrical activity under the electrode area. In this model, the limb is considered as a multilayered cylinder where muscle, adipose and skin tissues are described. Therefore, we propose a mechanical model described at the Motor Unit (MU) scale. The mechanical outcomes (muscle force, stiffness and deformation) are determined from the same voluntary command of the PNS, and is based on the Huxley sliding filaments model upscale at the MU scale using the distribution-moment theory proposed by Zahalak. This model is validated with force profile recorded from a subject implanted with an electrical stimulation device. Finally, we proposed three applications of the proposed models to illustrate their reliability and usefulness. A global sensitivity analysis of the statistics computed over the sEMG signals according to variation of the HD-sEMG electrode grid is performed. Then, we proposed in collaboration a new HDsEMG/force relationship, using personalized simulated data of the Biceps Brachii from the electrical model and a Twitch based model to estimate a specific force profile corresponding to a specific sEMG sensor network and muscle configuration. To conclude, a deformableelectro-mechanicalmodelcouplingthetwoproposedmodelsisproposed. This deformable model updates the limb cylinder anatomy considering isovolumic assumption and respecting incompressible property of the muscle.

Three novel mutations (G159D, L29Q and E59D/D75Y) in cardiac troponin C (CTnC) associate their clinical outcomes with a given cardiomyopathy. Current paradigms propose that sarcomeric mutations associated with dilated cardiomyopathy (DCM) decrease the myofilament calcium sensitivity while those associated with hypertrophic (HCM) cardiomyopathy increase it. Therefore, we incorporated the mutant CTnCs into skinned cardiac muscle in order to determine if their effects on the calcium regulation of tension and ATPase activity coincide with the current paradigms and phenotypic outcomes. This required the development of new calculator programs to solve complex ionic equilibria to more accurately buffer and expand the free calcium range of our test solutions. In accordance with the DCM paradigms, our result show that G159D and E59D/D75Y CTnC decrease the myofilament calcium sensitivity and force generating capabilities which would likely increase the rate of muscle relaxation and weaken the contractile force of the myocardium. Alternatively, the lack of myofilament change from L29Q CTnC (associated with HCM) may explain why the only proband is seemingly unaffected. Notably, the changes in the calcium sensitivity of tension (in fibers) do not necessarily occur in the isolated CTnC and vice versa. These counter-intuitive findings are justified through a transition in calcium affinity occurring at the level of cardiac troponin (CTn) and higher, implying that the true effects of these mutations become apparent as the hierarchal level of the myofilament increases. Despite these limitations, the regulated thin filament (RTF) retains its role as the calcium regulatory unit and best indicates a mutation's ability to sensitize (+) or desensitize (-) the muscle to calcium. Since multiple forms of cardiomyopathies exist, the identification of new drugs that sensitize (+) or desensitize (-) the calcium sensitivity could potentially reverse (+ or -) these aberrant changes in myofilament sensitivity. Therefore, we have developed an RTF mediated High Throughput Screening assay to identify compounds in libraries of molecules that can specifically modulate the calcium sensitivity of cardiac contraction. The knowledge gained from these studies will help us and others to uncover new pharmacological agents for the investigation and treatments of cardiomyopathies, hypertension and other forms of cardiovascular diseases.

Diss. (sammanfattning) Göteborg : Univ., 2003. / Härtill 5 uppsatser. Del av upplagan utan uppsatser.

This thesis presents the development and in-vivo applications of wearable and portable devices for the investigation of light interaction with tissue involved in Photodynamic therapy (PDT) and during contraction of muscles. A hand-held device and a clinical method were developed for time course in-vivo imaging of the fluorescence of the photosensitizer Protoporphyrin IX (PpIX) in healthy and diseased skin with the aim to guide improvement of PDT protocols. The device was used in a small clinical study on 11 healthy volunteers and 13 patients diagnosed with non-melanoma skin cancer (NMSC). Two types of PpIX precursors were administered: Ameluz gel and Metvix® cream. The fluorescence was imaged with a 10 minute time step over three hours which was the recommended metabolism time before commencing PDT treatment at Ninewells Hospital, Dundee. The fluorescence time course was calculated by integrating the areas with the highest intensity. The fluorescence continued to grow in all subjects during the three hours. The time course varied between individuals. There was no statistical significance between either healthy volunteers or patients in Ameluz vs Metvix® groups; nor was there statistical difference between the three lesions groups (Actinic keratosis (AK) Ameluz vs AK Metvix® vs Basal cell carcinoma (BCC) Metvix®). The p-value was larger than 0.05 in a two sample t-test with unequal variances for all the groups. However, there was strong body site dependence between the head & neck compared to the lower leg & feet, or the trunk & hands body site groups (p-value < 0.01). One of the possible explanations for this was temperature and vasculature variation in skin at different body sites: the temperature is higher and the vasculature structure is denser at the head and the neck compared to the lower leg or the trunk. The temperature was not measured during the study. So in order to support this hypothesis, typical skin temperatures at the lesion sites were taken from the IR thermal images of healthy skin available in literature. PpIX fluorescence had a positive correlation to temperature. If this hypothesis is true, it will be highly important to PDT treatment. Increasing the temperature could speed up the metabolism and reduce the waiting time before starting the treatment; ambient temperature should be taken into account for daylight PDT; cooling air as pain management should be administered with caution. Potential improvements for wearable PDT light sources were investigated by modelling light transport in skin for the current LED-based Ambulight PDT device, a commercial OLED for future devices and a directional OLED developed in the group. The optical models were implemented in commercial optical software (with intrinsic Monte Carlo ray tracing and Henyey-Greenstein scattering approximation) which was validated on diffuse reflectance and transmittance measurements using in-house made tissue phantoms. The modelling was applied to investigate the benefits from diffusive and forward scattering properties of skin on light transmission in treatment light sources. 1 mm thick skin can only compensate approximately 10% of non-uniform irradiance. It means that uniform illumination is crucial for the treatment light sources. Forward scattering in skin showed a 10% improved light transmission from a collimated emission compared to a wide angle Lambertian emission. However, depth-dependent transmission measurements of directional vs Lambertian emission from organic light emitting films (a nano-imprinted grating was fabricated to provide directional emission in one of the films), collimated vs diffused HeNe laser light through fresh porcine skin did not show the expected improvement. This could be explained by skin roughness which was previously found to change the optical properties and may also affect light coupling. The modelling was applied to guide an optical design of another wearable device – a muscle contraction sensor. Muscle is fibrous and because of that scatters light differently in different directions. The sensor detects the change in backscattered light in parallel and perpendicular directions with respect to muscle fibres. The sensor was implemented on a wearable bandage on fully flexible substrate with flexible OLED and organic photodiodes. The major advantages of organic optoelectronic sensing compared to conventional electromyography (EMG) sensors are the ability to distinguish two types of contractions (isotonic and isometric), insensitivity to electromagnetic interference and the absence of an immune response due to non-invasive electrode-free sensing. Optical modelling was performed to understand the operation of the sensor. A 3D anisotropic optical model of scattering in muscle was created by geometrical manipulations with the standard Henyey-Greenstein scattering volumes. The penetration depth from the Super Yellow OLED was found to be 20-25 mm; the optimal separation between the source and the detector was found to be 20 mm. This distance provided a still detectable signal along with the best discrimination between the two backscatterings. When a 2 mm thick layer of skin and a 2 mm thick layer of adipose tissue were added to the model, the signal was hugely diffused. The discrimination between the two backscatterings decreased by three orders of magnitude, the penetration depth in muscle was reduced, and the intensity of the signal dropped down but was still detectable. With 5 mm thick adipose tissue and 2 mm thick skin the signal was too diffused and interacted with very shallow layers of muscle which approached the limits of the optical sensing of muscle activity.

No / When descending stairs bodyweight becomes supported on a single limb while the forwards-reaching contralateral limb is lowered in order to make contact with the step below. This is associated with lowering of the centre of mass (CoM), which in order to occur in a controlled manner, requires increased ankle and knee joint torque production relative to that in overground walking. We have previously shown that when descending steps or stairs older people operate at a higher proportion of their maximum eccentric capacity and at, or in excess of the maximum passive reference joint range of motion. This suggests they have reduced and/or altered control over their CoM and we hypothesised that this would be associated with alterations in muscle activity patterns and in the CoM vertical acceleration and velocity profiles during both the lowering and landing phases of stair descent. 15 older (mean age 75 years) and 17 young (mean age 25 years) healthy adults descended a 4-step staircase, leading with the right limb on each stair, during which CoM dynamics and electromyographic activity patterns for key lower-limb muscles were assessed. Maximum voluntary eccentric torque generation ability at the knee and ankle was also assessed. Older participants compared to young participants increased muscle co-contraction relative duration at the knee and ankle of the trailing limb so that the limb was stiffened for longer during descent. As a result older participants contacted the step below with a reduced downwards CoM velocity when compared to young participants. Peak downwards and peak upwards CoM acceleration during the descent and landing phases respectively, were also reduced in older adults compared to those in young participants. In contrast, young participants descended quickly onto the step below but arrested their downward CoM velocity sooner following landing; a strategy that was associated with longer relative duration lead-limb plantar flexor activity, increased peak upwards CoM acceleration, and a reduced landing duration. These results suggest that a reduced ability to generate high eccentric torque at the ankle in the forward reaching limb is a major factor for older participants adopting a cautious movement control strategy when descending stairs. The implications of this CoM control strategy on the incidences of falling on stairs are discussed.

CNPq / Introdução: As pessoas com lesão medular (LM) podem ter seus músculos paralisados ativados por meio da estimulação elétrica funcional (FES) sobre vias neurais presentes próximas à pele. Estas estimulações elétricas são importantes para a recuperação do trofismo neuromuscular ou durante o controle de movimento por próteses neurais. No entanto, ao longo da aplicação da FES, a fadiga ocorre, diminuindo a eficiência da contração, principalmente devido à hipotrofia neuromuscular presente nessa população. A aquisição da vibração das fibras musculares como indicador de fadiga é registrada por meio da técnica de mecanomiografia (MMG), que não sofre interferências elétricas decorrentes da aplicação da FES. Objetivo: Caracterizar a vibração do músculo reto femoral durante protocolo de fadiga neuromuscular eletricamente evocada em pessoas com lesão medular completa. Método: 24 membros (direito e esquerdo) de 15 participantes (idade: 27±5 anos) do sexo masculino (A e B na American Spinal Injury Impairment Scale) foram selecionados. Um estimulador elétrico operando como fonte de tensão, desenvolvido especialmente para pesquisa, foi configurado com: freqüência de pulso em 1 kHz (20% de ciclo de trabalho) e trem de pulsos (modulação) em 70 Hz (20% período ativo). O sinal triaxial [X (transversal), Y (longitudinal) e Z (perpendicular)] da MMG foi processado com filtro Butterworth de terceira ordem e banda passante entre 5 e 50 Hz. Previamente ao protocolo, a tensão de saída do estimulador foi incrementada (~3 V/s evitando-se a adaptação/habituação dos motoneurônios) até alcançar a extensão máxima eletricamente estimulada (EMEE) da articulação do joelho. Uma célula de carga foi usada para registrar a resposta de força, onde após a sua colocação, a intensidade da FES necessária para alcançar a EMEE foi aplicada e registrada pela célula de carga como 100% da força (F100%). Durante o protocolo de fadiga neuromuscular, a intensidade do estímulo foi incrementada durante o controle para manter a força em F100%. Quatro instantes (I - IV) foram selecionados entre F100% e a incapacidade da FES manter a resposta de força acima de 30% (F30%). O sinal foi processado nos domínios temporal (energia), espectral (frequência mediana) e wavelet (temporal-espectral com doze bandas de frequência entre 5 e 53 Hz). Os dados extraídos foram normalizados pelo instante inicial (I) gerando unidades arbitrárias (u.a.), e testados com estatística não paramétrica. Resultados: A frequência mediana não apresentou significância estatística. Em relação aos eixos de deslocamento da MMG, o eixo transversal mostrou o maior número de resultados estatisticamente significantivos. A energia da vibração das fibras musculares (domínio temporal) indicou diminuição entre os instantes I (músculo fresco) e II (pré-fadiga), como também entre os instantes I e IV (fadigado) com redução significativa. O domínio wavelet teve como foco o eixo transversal, especialmente as bandas de frequência de 13, 16, 20, 25 e 35 Hz, por terem indicado redução significativa durante a fadiga neuromuscular; principalmente, a banda de 25 Hz, que indicou redução significativa entre o instante I (valor da mediana dos dados de 0,53 u.a.) e os demais instantes [II (0,30 u.a), III (0,28 u.a.) e IV (0,24 u.a.)]. Conclusão: A fadiga neuromuscular é caracterizada pela redução da energia do sinal no eixo de deslocamento transversal (X) da vibração do músculo reto femoral, em pessoas com lesão medular completa, tanto no domínio temporal quanto principalmente no domínio wavelet, sendo a banda de frequência de 25 Hz a mais relevante, porque sua energia diminui com a ocorrência da fadiga neuromuscular. Estes achados abrem a possibilidade de aplicação em sistemas de malha fechada durante procedimentos de reabilitação física utilizando FES ou no controle de próteses neurais. / Introduction: People with spinal cord injury (SCI) may have the paralyzed muscles activated through functional electrical stimulation (FES) on neural pathways present below the skin. These electrical stimulations are important to restore the neuromuscular trophism or during the movement control using neural prostheses. However, prolonged FES application causes fatigue, which decreases the contraction strength, mainly due the neuromuscular hypotrophy in this population. The acquisition of myofibers’ vibration is recognized by mechanomyography (MMG) system and does not suffer electrical interference from the FES system. Objective: To characterize the rectus femoris muscle vibration during electrically evoked neuromuscular fatigue protocol in complete spinal cord injury subjects. Methods: As sample, 24 limbs (right and left) from 15 male participants (age: 27±5 y.o.) and ranked as A and B according to American Spinal Injury Impairment Scale) were selected. An electrical stimulator operating as voltage source, specially developed for research, was configured as: pulse frequency set to 1 kHz (20% duty cycle) and burst (modulating) frequency set to 70 Hz (20% active period). The triaxial [X (transverse), Y (longitudinal) and Z (perpendicular)] MMG signal of rectus femoris muscle was processed with a third-order 5-50 Hz bandpass Butterworth filter. A load cell was used to register the force. The stimulator output voltage was increased (~3 V/s to avoid motoneuron adaptation/habituation) until the maximal electrically-evoked extension (MEEE) of the knee joint. After the load cell placement, the stimuli magnitude required to reach MEEE was applied and registered by the load cell as muscular F100% response. Stimuli intensity was increased during the control to keep the force in F100%. Four instants (I - IV) were selected from F100% up to the inability to keep the FES response force above 30% (F30%). The signal was processed in temporal (energy), spectral (median frequency) and wavelet (temporal-spectral with twelve band frequencies between 5 and 53 Hz) domains. All data were normalized by initial instant, creating arbitrary units (a.u.), and non-parametric tests were applied. Results: The median frequency did not show statistical significance. Regarding the MMG axes, the transverse axis showed most statistical differences. The MMG energy (temporal domain) indicates the decrease between the instants I (unfatigued) and II (pre-fatigue), as well as instants I and IV (fatigued). The wavelet domain focused on the transverse axis, especially on 13, 16, 20, 25 and 35 Hz frequency bands, for having shown significant reduction proven during neuromuscular fatigue. In focus on 25 Hz band frequency that showed a constant decrease between instants I (median value from data de 0.53 a.u.) with subsequent instants [II (0.30 a.u.), III (0.28 a.u.) and IV (0.24 a.u.). Conclusion: Neuromuscular fatigue is characterized by energy decrease in MMG X-axis (transverse) signal of vibration on the rectus femoris muscle for complete spinal cord injured subjects, in the temporal domain but mainly in the wavelet domain. The 25 Hz is the most important band frequency because its energy decreases with neuromuscular fatigue. These findings open the possibility of application in closed-loop systems during physical rehabilitation procedures using FES or in the control of neural prostheses.

CNPq / Introdução: As pessoas com lesão medular (LM) podem ter seus músculos paralisados ativados por meio da estimulação elétrica funcional (FES) sobre vias neurais presentes próximas à pele. Estas estimulações elétricas são importantes para a recuperação do trofismo neuromuscular ou durante o controle de movimento por próteses neurais. No entanto, ao longo da aplicação da FES, a fadiga ocorre, diminuindo a eficiência da contração, principalmente devido à hipotrofia neuromuscular presente nessa população. A aquisição da vibração das fibras musculares como indicador de fadiga é registrada por meio da técnica de mecanomiografia (MMG), que não sofre interferências elétricas decorrentes da aplicação da FES. Objetivo: Caracterizar a vibração do músculo reto femoral durante protocolo de fadiga neuromuscular eletricamente evocada em pessoas com lesão medular completa. Método: 24 membros (direito e esquerdo) de 15 participantes (idade: 27±5 anos) do sexo masculino (A e B na American Spinal Injury Impairment Scale) foram selecionados. Um estimulador elétrico operando como fonte de tensão, desenvolvido especialmente para pesquisa, foi configurado com: freqüência de pulso em 1 kHz (20% de ciclo de trabalho) e trem de pulsos (modulação) em 70 Hz (20% período ativo). O sinal triaxial [X (transversal), Y (longitudinal) e Z (perpendicular)] da MMG foi processado com filtro Butterworth de terceira ordem e banda passante entre 5 e 50 Hz. Previamente ao protocolo, a tensão de saída do estimulador foi incrementada (~3 V/s evitando-se a adaptação/habituação dos motoneurônios) até alcançar a extensão máxima eletricamente estimulada (EMEE) da articulação do joelho. Uma célula de carga foi usada para registrar a resposta de força, onde após a sua colocação, a intensidade da FES necessária para alcançar a EMEE foi aplicada e registrada pela célula de carga como 100% da força (F100%). Durante o protocolo de fadiga neuromuscular, a intensidade do estímulo foi incrementada durante o controle para manter a força em F100%. Quatro instantes (I - IV) foram selecionados entre F100% e a incapacidade da FES manter a resposta de força acima de 30% (F30%). O sinal foi processado nos domínios temporal (energia), espectral (frequência mediana) e wavelet (temporal-espectral com doze bandas de frequência entre 5 e 53 Hz). Os dados extraídos foram normalizados pelo instante inicial (I) gerando unidades arbitrárias (u.a.), e testados com estatística não paramétrica. Resultados: A frequência mediana não apresentou significância estatística. Em relação aos eixos de deslocamento da MMG, o eixo transversal mostrou o maior número de resultados estatisticamente significantivos. A energia da vibração das fibras musculares (domínio temporal) indicou diminuição entre os instantes I (músculo fresco) e II (pré-fadiga), como também entre os instantes I e IV (fadigado) com redução significativa. O domínio wavelet teve como foco o eixo transversal, especialmente as bandas de frequência de 13, 16, 20, 25 e 35 Hz, por terem indicado redução significativa durante a fadiga neuromuscular; principalmente, a banda de 25 Hz, que indicou redução significativa entre o instante I (valor da mediana dos dados de 0,53 u.a.) e os demais instantes [II (0,30 u.a), III (0,28 u.a.) e IV (0,24 u.a.)]. Conclusão: A fadiga neuromuscular é caracterizada pela redução da energia do sinal no eixo de deslocamento transversal (X) da vibração do músculo reto femoral, em pessoas com lesão medular completa, tanto no domínio temporal quanto principalmente no domínio wavelet, sendo a banda de frequência de 25 Hz a mais relevante, porque sua energia diminui com a ocorrência da fadiga neuromuscular. Estes achados abrem a possibilidade de aplicação em sistemas de malha fechada durante procedimentos de reabilitação física utilizando FES ou no controle de próteses neurais. / Introduction: People with spinal cord injury (SCI) may have the paralyzed muscles activated through functional electrical stimulation (FES) on neural pathways present below the skin. These electrical stimulations are important to restore the neuromuscular trophism or during the movement control using neural prostheses. However, prolonged FES application causes fatigue, which decreases the contraction strength, mainly due the neuromuscular hypotrophy in this population. The acquisition of myofibers’ vibration is recognized by mechanomyography (MMG) system and does not suffer electrical interference from the FES system. Objective: To characterize the rectus femoris muscle vibration during electrically evoked neuromuscular fatigue protocol in complete spinal cord injury subjects. Methods: As sample, 24 limbs (right and left) from 15 male participants (age: 27±5 y.o.) and ranked as A and B according to American Spinal Injury Impairment Scale) were selected. An electrical stimulator operating as voltage source, specially developed for research, was configured as: pulse frequency set to 1 kHz (20% duty cycle) and burst (modulating) frequency set to 70 Hz (20% active period). The triaxial [X (transverse), Y (longitudinal) and Z (perpendicular)] MMG signal of rectus femoris muscle was processed with a third-order 5-50 Hz bandpass Butterworth filter. A load cell was used to register the force. The stimulator output voltage was increased (~3 V/s to avoid motoneuron adaptation/habituation) until the maximal electrically-evoked extension (MEEE) of the knee joint. After the load cell placement, the stimuli magnitude required to reach MEEE was applied and registered by the load cell as muscular F100% response. Stimuli intensity was increased during the control to keep the force in F100%. Four instants (I - IV) were selected from F100% up to the inability to keep the FES response force above 30% (F30%). The signal was processed in temporal (energy), spectral (median frequency) and wavelet (temporal-spectral with twelve band frequencies between 5 and 53 Hz) domains. All data were normalized by initial instant, creating arbitrary units (a.u.), and non-parametric tests were applied. Results: The median frequency did not show statistical significance. Regarding the MMG axes, the transverse axis showed most statistical differences. The MMG energy (temporal domain) indicates the decrease between the instants I (unfatigued) and II (pre-fatigue), as well as instants I and IV (fatigued). The wavelet domain focused on the transverse axis, especially on 13, 16, 20, 25 and 35 Hz frequency bands, for having shown significant reduction proven during neuromuscular fatigue. In focus on 25 Hz band frequency that showed a constant decrease between instants I (median value from data de 0.53 a.u.) with subsequent instants [II (0.30 a.u.), III (0.28 a.u.) and IV (0.24 a.u.). Conclusion: Neuromuscular fatigue is characterized by energy decrease in MMG X-axis (transverse) signal of vibration on the rectus femoris muscle for complete spinal cord injured subjects, in the temporal domain but mainly in the wavelet domain. The 25 Hz is the most important band frequency because its energy decreases with neuromuscular fatigue. These findings open the possibility of application in closed-loop systems during physical rehabilitation procedures using FES or in the control of neural prostheses.