Дисертації з теми "Skeletal muscular system"

ZEB1 is a transcription factor best known for its role in cancer progression and metastasis. It is also expressed during embryonic development of different tissues although its function and mechanism of action have not always been elucidated. In this dissertation I show that ZEB1 is involved in muscle differentiation during embryonic development and it is also required for muscle response after injury and regeneration. We found that, in the nucleus of myoblasts, ZEB1 represses muscle differentiation genes through direct binding to G/C-centered E-boxes present in the regulatory regions of muscle differentiation genes. Albeit to different degrees depending on the target gene, transcriptional repression of these genes by ZEB1 is mediated by its recruitment of the corepressor CtBP. Binding of ZEB1 to E-boxes in differentiation genes displaces MyoD and prevents their transcriptional activation during the myoblast stage. As myoblasts fuse, MyoD displaces ZEB1 from its DNA binding sites and differentiation proceeds. Knockdown of Zeb1 induces muscle differentiation genes, thus accelerating the formation of myotubes. Muscle regeneration after damage depends on a timely regulated transition from pro- to anti-inflammatory signals. Injury of Zeb1-deficient mice results in increased recruitment of inflammatory macrophages and expression of pro-inflammatory cytokines, which delays the regenerative process. Adult muscle regeneration relies on a pool of functional SCs and we show that Zeb1-deficient SCs undergo premature activation after isolation and culture by downregulating Pax7 and quiescence-associated genes (Foxo3, Hes genes) and upregulating Myod1. Moreover, its regenerative potential when transplanted into mdx hosts is reduced compared to wild-type SCs and exhibit increased senescence in culture. These results establish ZEB1 as an important potential regulator of muscle differentiation and regeneration by modulating inflammatory response and SC myogenic progression in response to injury. They also set ZEB1 as a potential therapeutic target in muscle dystrophies or following muscle insult.
ZEB1 és un factor de transcripció conegut pel seu paper en progressió tumoral i metàstasi. També s’expressa durant el desenvolupament embrionari de diferents teixits tot i que la seva funció i mecanisme d’acció encara no han estat establerts. En aquesta tesi mostro que ZEB1 està implicat en la diferenciació muscular durant el desenvolupament embrionari i que es necessari en la resposta al dany i regeneració muscular. Hem trobat que, en els nuclis dels mioblasts, ZEB1 reprimeix gens de diferenciació muscular per unió directa a seqüències “E-box” amb nucleòtids G/C en posició central en les seves regions reguladores. Encara que en diferents graus, depenent del gen diana, la repressió exercida per ZEB1 es fa mitjançant el reclutament del seu corepressor CtBP. La unió de ZEB1 a aquestes “E-boxes” desplaça MyoD evitant la seva activació transcripcional. Un cop els mioblasts es fusionen, MyoD desplaça ZEB1 de la seva unió a l’ADN donant lloc al procés de diferenciació. D’aquesta manera, la inhibició de Zeb1 indueix els gens de diferenciació muscular accelerant la formació de miotubs. La regeneració desprès del dany muscular depèn de la transició de senyals proinflamatoris a antiinflamatoris. La lesió muscular de ratolins deficients per Zeb1 produeix un elevat nombre de macròfags inflamatoris i l’expressió de citocines pro-inflamatories que retarden el procés regeneratiu. La regeneració del teixit muscular adult requereix la participació d’una població de cèl·lules satèl·lit funcionals. Els nostres resultats demostren que les cèl·lules satèl·lit deficients per Zeb1 s’activen precoçment un cop aïllades i posades en cultiu. Aquesta activació succeeix per la inhibició de Pax7 i de gens associats a la quiescència d’aquestes cèl·lules (Foxo3, Hes) i la activació de Myod1. A més a més, presenten una més alta senescència i la seva capacitat regenerativa és reduïda quan es trasplanten en ratolins mdx en comparació a les wild-type. Aquests resultats situen ZEB1 com un important regulador de la diferenciació i la regeneració muscular per modulació de la resposta inflamatòria i de la progressió de les cèl·lules satèl·lit en la resposta al dany muscular. També suggereixen ZEB1 com una potencial diana terapèutica en distròfies musculars o en resposta a la lesió del múscul esquelètic.

Introdução: Pacientes com cardiopatia chagásica têm hiperatividade do sistema nervoso simpático, piorando o prognóstico destes pacientes. Estão bem estabelecidos os benefícios do treinamento físico aeróbico (TF) no controle autonômico cardiovascular e na musculatura esquelética de pacientes com cardiopatia e disfunção ventricular. A hipótese da tese seria que o TF melhorasse a função autonômica cardiovascular e a estrutura e metabolismo muscular de pacientes com cardiopatia chagásica crônica (CCC) mesmo com função sistólica preservada, tendo em vista que parte destes pacientes evolui para a forma dilatada com disfunção ventricular e suas graves consequências. Objetivo: Avaliar o efeito do TF no controle autonômico cardiovascular e no tecido muscular esquelético em pacientes com CCC e função sistólica preservada. Métodos: Foram incluídos pacientes com duas reações sorológicas positivas para a doença de Chagas, alterações eletrocardiográficas, fração de ejeção do ventrículo esquerdo >= 55% e idade entre 30 e 60 anos. Vinte e quatro pacientes foram submetidos à primeira série de avaliações e foram randomizados em dois grupos: doze pacientes com CCC e função ventricular sistólica preservada submetidos ao TF além do seguimento clínico (ChT) e doze pacientes com CCC e função ventricular sistólica preservada não submetidos ao TF, apenas ao seguimento clínico (ChNT). Após quatro meses, oito pacientes finalizaram o protocolo de treinamento físico (ChT, n=08) e dez pacientes finalizaram o seguimento clínico (ChNT, n=10). A atividade nervosa simpática muscular (ANSM) foi avaliada pela técnica de microneurografia e o fluxo sanguíneo muscular (FSM) pela técnica de pletismografia de oclusão venosa. Variabilidade da frequência cardíaca e da pressão arterial foram analisadas utilizando sinais da frequência cardíaca captadas pelo eletrocardiograma e sinais da pressão arterial captados pelo finometer. A sensibilidade barorreflexa cardíaca foi avaliada com infusão de drogas vasotivas. A capacidade funcional foi avaliada pelo teste cardiopulmonar. A biópsia do músculo vasto-lateral foi realizada para as análises histológicas das fibras musculares e para avaliação da expressão gênica de Atrogin-1 e MuRF-1. O programa de TF foi realizado durante quatro meses, constando de 3 sessões semanais supervisionadas com duração aproximada de 60 minutos. Resultados: Como marcadores de TF, houve redução da frequência cardíaca de repouso e aumento do consumo de oxigênio pico. O TF diminuiu a hiperatividade simpática, colaborando para o aumento do FSM. O treinamento físico reduziu tanto a ANSM, quanto a atividade simpática cardíaca e vasomotora, e melhorou a sensibilidade barorreflexa cardíaca. A redução da ANSM esteve associada a redução da hiperatividade cardiovascular, melhora da sensibilidade barorreflexa cardíaca e redução da expressão gênica de Atrogin-1 e MuRF-1. Após período de quatro meses, o grupo ChT apresentou menor expressão gênica de Atrogin-1 em relação ao grupo ChNT. Conclusão: O TF provocou expressiva melhora na disfunção autonômica, no FSM e na capacidade funcional de pacientes com CCC e função sistólica preservada. Adicionalmente, a redução da ANSM esteve associada a melhora da sensibilidade barorreflexa cardíaca, redução do tônus simpático cardiovascular e redução da expressão gênica de Atrogin-1 e MuRF-1, genes envolvidos na atrofia muscular
[thesis]. São Paulo: Faculdade de Medicina, Universidade de São Paulo; 2017. Background: Patients with chagasic cardiomyopathy have sympathetic nervous system hyperactivity, worsening the prognosis of these patients. The benefits of aerobic training (ET) in cardiovascular autonomic control and skeletal muscle of heart failure patients are well established. The thesis hypothesis was that ET improves cardiovascular autonomic function and structure and metabolism muscle in chronic chagasic cardiopathy (CCC) patients even though preserved systolic function, considering that part of these patients develop the dilated form with ventricular dysfunction and its serious consequences. Objectives: To evaluate the effects of ET on cardiovascular autonomic control and skeletal muscle tissue in CCC patients and preserved systolic function. Methods: Patients with two positive serological reactions for Chagas disease, electrocardiographic alterations, left ventricular ejection fraction >= 55% and age between 30 and 60 years were included. Twenty-four patients underwent the first stage of evaluations and were randomized into two groups: Twelve CCC patients and preserved systolic ventricular function submitted to ET in addition to clinical follow-up (ET group) and twelve CCC patients and preserved systolic ventricular function submitted to only clinical follow-up not submitted to ET (NoET group). After four months, eight patients completed the ET protocol (ET, n = 08) and ten patients completed clinical follow-up (NoET, n = 10). Muscular sympathetic nerve activity (MSNA) was measured using microneurography technique and muscle blood flow (MBF) by the venous occlusion plethysmography technique. Heart rate and blood pressure variability were analyzed using heart rate signals captured by the electrocardiogram and blood pressure signals captured by the finometer. Cardiac baroreflex sensitivity was evaluated by infusion of vasoactive drugs. Functional capacity was determined by cardiopulmonary exercise test. Vastus lateralis muscle biopsy was performed for the histological analysis of muscle fibers and for the Atrogin-1 and MuRF-1 gene expression evaluation. ET program consisted of three 60-minute exercise sessions per week for four months. Results: As ET markers, there was a reduction in resting heart rate and an increase in peak oxygen consumption. ET reduced the sympathetic hyperactivity, contributing to the increase of the MBF. ET reduced both MSNA, as well as cardiac and vasomotor sympathetic activity, and improved cardiac baroreflex sensitivity. Reduction of MSNA was associated with a reduction in cardiovascular hyperactivity, improved cardiac baroreflex sensitivity, and reduced Atrogin-1 and MuRF-1 gene expression. After the four-month period, the ET group presented lower Atrogin-1 gene expression than the NoET group. Conclusion: ET improved significantly autonomic dysfunction, MBF and functional capacity of CCC patients and preserved systolic function. In addition, the reduction of ANSM was associated with improved cardiac baroreflex sensitivity, reduced sympathetic cardiovascular tone, and reduced Atrogin-1 and MuRF-1 gene expression, genes involved in muscle atrophy

Thesis (MSc (Physiological Sciences))--Stellenbosch University, 2008.
Introduction: Skeletal muscle atrophy is a mitigating complication that is characterized by a reduction in muscle fibre cross-sectional area as well as protein content, reduced force, elevated fatigability and insulin resistance. It seems to be a highly ordered and regulated process and signs of this condition are often seen in inflammatory conditions such as cancer, AIDS, diabetes and chronic heart failure (CHF). It has long been understood that an imbalance between protein degradation (increase) and protein synthesis (decrease) both contribute to the overall loss of muscle protein. Although the triggers that cause atrophy are different, the loss of muscle mass in each case involves a common phenomenon that induces muscle proteolysis. It is becoming evident that interactions among known proteolytic systems (ubiquitin-proteosome) are actively involved in muscle proteolysis during atrophy. Factors such as TNF-α and ROS are elevated in a wide variety of chronic inflammatory diseases in which skeletal muscle proteolysis presents a lethal threat. There is an increasing body of evidence that implies TNF-α may play a critical role in skeletal muscle atrophy in a number of clinical settings but the mechanisms mediating its effects are not completely understood. It is also now apparent that the transcription factor NF-κB is a key intracellular signal transducer in muscle catabolic conditions. This study investigated the various proposed signalling pathways that are modulated by increasing levels of TNF-α in a skeletal muscle cell line, in order to synthesize our current understanding of the molecular regulation of muscle atrophy. Materials and Methods: L6 (rat skeletal muscle) cells were cultured under standard conditions where after reaching ± 60-65% confluency levels, differentiation was induced for a maximum of 8 days. During the last 2 days, myotubes were incubated with increasing concentrations of recombinant TNF-α (1, 3, 6 and 10 ng/ml) for a period of 40 minutes, 24 and 48 hours. The effects of TNF-α on proliferation and cell viability were measured by MTT assay and Trypan Blue exclusion technique. Morphological assessment of cell death was conducted using the Hoechst 33342 and Propidium Iodide staining method. Detection of apoptosis was assessed by DNA isolation and fragmentation assay. The HE stain was used for the measurement of cell size. In order to determine the source and amount of ROS production, MitoTracker Red CM-H2 X ROS was utilised. Ubiquitin expression was assessed by immunohistochemistry. PI3-K activity was calculated by using an ELISA assay and the expression of signalling proteins was analysed by Western Blotting using phospho-specific and total antibodies. Additionally, the antioxidant Oxiprovin was used to investigate the quantity of ROS production in TNF-α-induced muscle atrophy. Results and Discussion: Incubation of L6 myotubes with increasing concentrations of recombinant TNF-α revealed that the lower concentrations of TNF-α used were not toxic to the cells but data analysis of cell death showed that 10 ng/ml TNF-α induced apoptosis and necrosis. Long-term treatment with TNF-α resulted in an increase in the upregulation of TNF- α receptors, specifically TNF-R1. The transcription factors NF-κB and FKHR were rapidly activated thus resulting in the induction of the ubiquitin-proteosome pathway. Activation of this pathway produced significant increases in the expression of E3 ubiquitin ligases MuRF-1 and MAFbx. Muscle fibre diameter appeared to have decreased with increasing TNF-α concentrations in part due to the suppressed activity of the PI3-K/Akt pathway as well as significant reductions in differentiation markers. Western blot analysis also showed that certain MAPKs are activated in response to TNF-α. No profound changes were observed with ROS production. Finally, the use Oxiprovin significantly lowered cell viability and ROS production. These findings suggest that TNF-α may elicit strong catabolic effects on L6 myotubes in a dose and time dependent manner. Conclusion: These observations suggest that TNF-α might have beneficial effects in skeletal muscle in certain circumstances. This beneficial effect however is limited by several aspects which include the concentration of TNF-α, cell type, time of exposure, culture conditions, state of the cell (disturbed or normal) and the cells stage of differentiation. The effect of TNF-α can be positive or negative depending on the concentration and time points analysed. This action is mediated by various signal transduction pathways that are thought to cooperate with each other. More understanding of these pathways as well as their subsequent upstream and downstream constituents is obligatory to clarify the central mechanism/s that control physiological and pathophysiological effects of TNF-α in skeletal muscle.

Introdução. A insuficiência cardíaca (IC) é caracterizada por exacerbação da atividade nervosa simpática muscular (ANSM), baixa tolerância ao esforço e dispneia. Além disso, é característico nessa população o desequilíbrio entre o anabolismo e catabolismo, favorecendo dessa maneira uma acentuada perda da massa magra muscular, o que agrava ainda mais a qualidade de vida nos pacientes com IC. Dentre as alterações anabólicas observadas na IC avançada destaca-se a diminuição dos hormônios GH, IGF-1 e testosterona. A testosterona, um importante hormônio para as características masculinizantes e na manutenção da massa muscular, apresenta acentuada redução com o avançar da doença. Esta perda da massa magra, leva ao processo de caquexia muscular e consequente atrofia, com diminuição da força e da capacidade funcional do paciente com IC. A reposição de testosterona nesses pacientes tem sido estudada e se mostra uma importante terapêutica para melhorar a capacidade funcional e força muscular. Porém, não se conhece claramente o papel deste tratamento medicamentoso sobre o processo anabólico muscular, bem como na melhora da composição corporal. O exercício físico como tratamento não medicamentoso tem sido amplamente recomendado na IC por reduzir a ANSM, melhorar o fluxo sanguíneo periférico, aumentar a força muscular e melhorar a qualidade de vida. Entretanto, a combinação das estratégias do exercício físico associado à terapia de reposição de testosterona, não é conhecido em pacientes com IC. Métodos. 24 pacientes com IC foram randomizados em 3 grupos: Treinamento (TR, n=9), Testosterona (T, n=8) e Treino+Testosterona (TRT, n=7). A ANSM foi avaliada pela técnica de Microneurografia. O fluxo sanguíneo do antebraço foi avaliado pela pletismografia de oclusão venosa. A composição corporal foi avaliada pela densitometria (DEXA). A biópsia do músculo vasto-lateral foi feita para avaliarmos a área de secção transversa da fibra e a tipagem de fibras musculares. A qualidade de vida foi avaliada pelo questionário de Minnesota. O treinamento físico aeróbio em bicicleta foi realizado 3 vezes por semana, com 40 minutos de exercício por sessão, pelo período de 4 meses. A reposição de testosterona foi realizada pela administração intramuscular de undecilato de testosterona pelo período de 4 meses. Resultados. Após 4 meses de intervenção, observamos restauração dos níveis de testosterona em todos os grupos. A ANSM reduziu nos grupos TR e TRT. Não houve aumento do fluxo sanguíneo entre os grupos. O consumo de oxigênio aumentou em todos os grupos, porém apenas o grupo TRT aumentou a potência máxima ao exercício. A massa magra apresentou aumento significativo apenas no grupo TRT. Não observamos mudança no conteúdo mineral ósseo entre os grupos. Apenas o grupo TRT aumentou de maneira significativa a área de secção transversa das fibras tipo I (oxidativas). A qualidade de vida melhorou apenas nos grupos TR e TRT. Conclusões. O exercício físico associado à terapia de reposição de testosterona se mostrou mais eficaz em reduzir a ANSM, aumentar a capacidade funcional, a força muscular, a massa magra com um importante aumento das fibras do tipo I. Nossos resultados enfatizam a importância do exercício físico em pacientes com IC e traz uma nova perspectiva com a associação da testosterona para pacientes com hipogonadismo
Introduction. Heart failure (HF) is characterized by exacerbation of muscle sympathetic nerve activity (MSNA), exercise intolerance and dyspnea. Furthermore, is characteristic in this population the imbalance between anabolism and catabolism which lead to loss of skeletal muscle mass worsening quality of life in HF patients. Prior studies have demonstrated decrease in anabolic hormones such as GH, IGF-1 and testosterone. Testosterone, an important hormone for masculinization feature and maintenance of muscle mass, shows sharp decline in advanced HF. Loss muscle mass leads to cachexia and atrophy which decrease strength and functional capacity in HF patients. Testosterone replacement in these patients has been studied and shows an important therapeutic to enhance functional capacity and muscle strength. However it is not known the role of this medical treatment on muscle anabolic process as well as on body composition. Physical exercise as a non-medication treatment has been widely recommended to reduce MSNA, enhance peripheral blood flow, increase muscle strength and improve quality of life. However, the combination of the strategies of physical exercise associated with testosterone replacement therapy is not known in HF patients. Methods. 24 HF patients were randomized in 3 groups: Training (TR, n=9), Testosterone (T, n=8) and Training+Testosterone (TRT, n=7). MSNA was recorded by microneurography technic. Forearm blood flow was evaluated by venous occlusion plethysmography. Body composition was measured by densitometry (DEXA). Muscle biopsy was done in vastus lateralis to evaluate the cross-sectional area and type of fibers. Quality of life was assessed by Minnesota living with heart failure questionnaire. Aerobic exercise training on a bicycle was performed 3 times per week, with 40 minutes of exercise per session, for a period of 4 months. Testosterone replacement was performed by intramuscular administration of testosterone undecylate for a period of 4 months. Results. After 4 months testosterone levels were restored in all groups. MSNA decreased in TR and TRT groups. There was no increase in blood flow between groups. Oxygen consumption increased in all groups, but only the TRT group showed increase in maximum power to exercise. Lean body mass increased significantly only in the TRT group. We did not observe changes in bone mineral content between groups. Only TRT group significantly increased the cross-sectional area of type I fibers (oxidative). The quality of life improved only in TR and TRT groups. Conclusions. Exercise training associated with testosterone replacement therapy was more effective in reducing MSNA, increase functional capacity, muscle strength, lean mass with a significant increase in type I fibers. Our results emphasize the importance of physical exercise in patients with HF and bring a new perspective to association testosterone for patients with hypogonadism

Introdução. A hiperativação nervosa simpática é característica marcante da insuficiência cardíaca. Estudos apontam alterações no controle ergorreflexo muscular (mecano e metaborreflexo) como mecanismos potenciais para explicar esta modificação autonômica. Os mecanorreceptores (fibras do grupo III), que são ativadas pelo aumento no tônus muscular e modulados por metabólitos da via das ciclooxigenases, encontram-se hipersensibilizadas na insuficiência cardíaca. Ao contrário, a sensibilidade dos metaborreceptores (fibras do grupo IV), que são ativados pelo acúmulo de metabólitos durante as contrações musculares e modulados pelos receptores TRPV1 e CB1, encontra-se diminuída na insuficiência cardíaca. Por outro lado, o treinamento físico tem se mostrado uma importante ferramenta no tratamento da insuficiência cardíaca. Ele reduz os níveis de atividade nervosa simpática muscular (ANSM) no repouso e durante o exercício em pacientes portadores desta síndrome. Dessa forma, neste estudo, nós testamos a hipótese de que o treinamento físico melhoraria o controle mecano e metaborreflexo da ANSM em pacientes com insuficiência cardíaca, e se essa melhora está associada às alterações na via das ciclooxigenases e na expressão dos receptores TRPV1 e CB1, respectivamente. Métodos. Pacientes com insuficiência cardíaca foram consecutivamente e aleatoriamente divididos em dois grupos: insuficiência cardíaca não treinado (ICNT, n=17) e insuficiência cardíaca treinado (ICT, n=17). A ANSM foi avaliada pela técnica de microneurografia e o fluxo sanguíneo muscular (FSM) pela pletismografia de oclusão venosa. A frequência cardíaca (FC) e a pressão arterial (PA) foram avaliadas por medida não invasiva a cada batimento (Finometer). Foi realizada biopsia muscular do vasto lateral para análise de expressão gênica. O treinamento físico aeróbio foi realizado em ciclo ergômetro, em intensidade moderada, por 40 minutos, três vezes por semana, durante 16 semanas. A sensibilidade mecanorreflexa foi calculada pelo delta absoluto entre o pico do exercício passivo, realizado na perna esquerda, e a média do registro basal. A sensibilidade metaborreflexa foi calculada pelo delta absoluto entre o 1º minuto de oclusão circulatória pós-exercício na perna esquerda e a média do registro basal. Resultados. O treinamento físico reduziu a ANSM e aumentou o FSM no repouso. O treinamento físico diminuiu significativamente as respostas de ANSM durante o exercício passivo no grupo ICT. As repostas de PA média também foram menores no grupo ICT quando comparado ao grupo ICNT. Não houve alterações significativas nas repostas de FC, PA sistólica, PA diastólica e FSM durante o exercício passivo no grupo ICT. Em relação à sensibilidade metaborreflexa, o treinamento físico aumentou expressivamente as respostas de ANSM no 1º minuto de oclusão circulatória no grupo ICT. As respostas de FC, PA e FSM não foram alteradas neste grupo. Não foram observadas alterações significativas nos controles mecano e metaborreflexo musculares no grupo ICNT. Além disso, o treinamento físico reduziu significativamente a expressão gênica da enzima COX-2 e do receptor EP4 e aumentou significativamente a expressão dos receptores TRPV1 e CB1 no grupo ICT. Não foram verificadas alterações significativas nas expressões gênicas do grupo ICNT. Conclusões. O treinamento físico normaliza os controles mecano e metaborreflexo da ANSM em pacientes com insuficiência cardíaca. Estas alterações podem estar associadas às alterações na expressão gênica da enzima COX-2 e receptor EP4, e dos receptores TRPV1 e CB1, respectivamente. Em conjunto, estes achados podem explicar, pelo menos em parte, a diminuição da atividade nervosa simpática e a melhora na tolerância aos esforços em pacientes com insuficiência cardíaca
Introduction. Sympathoexcitation is the hallmark of heart failure. Studies suggest changes in ergoreflex muscle control (mechanoreflex and metaboreflex) as potential mechanisms to explain this autonomic alteration in heart failure. Mechanoreceptors (group III fibers) that are activated by mechanical stimuli and modulated by cyclooxygenase pathway metabolites are hypersensitive in heart failure. In contrast, the sensitivity of metaboreceptors fibers (group IV) that are activated by increases in ischemic metabolites during muscle contractions and modulated by TRPV1 and CB1 receptors is blunted in heart failure. On the other hands, exercise training has been shown to be an important strategy in the treatment of heart failure. It reduces the levels of muscle sympathetic nerve activity (MSNA) at rest and during exercise in patients suffering of this syndrome. Thus, we tested the hypothesis that exercise training would improve the mechanoreflex and metaboreflex control of MSNA in heart failure patients. In addition, we investigated whether the improvement in the mechanoreflex and metaboreflex control is related to changes in the cyclooxygenase pathway and expression of TRPV1 and CB1 receptors, respectively. Methods. Patients with heart failure were consecutively and randomly divided into two groups: heart failure untrained (HFUT, n = 17) and heart failure exercise-trained (HFET, n = 17). MSNA was measured by microneurography technique and muscle blood flow (MBF) by venous occlusion plethysmography. Heart rate (HR) and blood pressure (BP) were assessed by noninvasive measure on a beat-to-beat basis (Finometer). Gene expression analysis was investigated by vastus lateralis muscle biopsy. Aerobic exercise training was performed on a cycle ergometer at moderate intensity, three 40-min session/wk for 16 weeks. Mechanoreflex sensitivity was evaluated by means the absolute difference in MSNA at peak passive exercise and baseline. Metaboreflex sensitivity was calculated by means the absolute difference in MSNA at 1st min after exercise period with muscle circulatory arrest and baseline. Results. Exercise training reduced MSNA and increased MBF. Exercise training significantly decreased MSNA responses during passive exercise. The mean BP response was lower in HFET group when compared to HFUT group. There were no significant changes in HR, systolic and diastolic BP and MBF responses during passive exercise in HFET group. Regarding metaboreflex sensitivity, exercise training significantly increased the MSNA responses at 1st minute of post exercise circulatory arrest. The responses of HR, BP and MBF were unchanged after exercise training. No significant changes were observed in mechanoreflex and metaboreflex control in the HFUT group. Furthermore, exercise training significantly reduced gene expression of COX-2 and EP4 receptor and significantly increased expression of TRPV1 and CB1 receptors. There were no significant changes in the gene expressions in the HFUT group. Conclusions. Exercise training improves mechanoreflex and metaboreflex control of MSNA in heart failure patients. These changes may be associated with changes in gene expression of COX-2 and EP4 receptor and TRPV1 and CB1 receptor, respectively. Together, these findings may explain, at least in part, the decrease in sympathetic nerve activity and the improvement in exercise tolerance in patients with heart failure

La mutagenèse chimique aléatoire par l’Ethyl-Nitroso-Urée (ENU), dont la puissance a été largement démontrée au cours de ces dernières décennies dans la création de modèles murins constitue un outil remarquable et essentiel en génomique fonctionnelle. Cette approche est en effet d’un apport considérable pour la compréhension de la fonction des gènes et de leur régulation et constitue un accélérateur pour identifier des éléments clés dans une voie de signalisation. Cette approche systématique, basée sur le criblage simultané d’un grand nombre de souris ne nécessite aucune connaissance préalable sur l’identité et la fonction des gènes étudiés. Les mutants sont identifiés au travers de cribles phénotypiques spécifiques, hiérarchisés et non-invasifs. L’identification du gène et de la mutation causale responsable du phénotype sont réalisés par un travail de cartographie génique en utilisant une série de marqueurs polymorphes ou par séquençage nouvelle génération. L’objectif de ma thèse a été de caractériser au niveau phénotypique et moléculaire, 3 lignées murines indépendantes issues de deux cribles ENU, un crible récessif et un crible dominant sensibilisé. Le premier crible visait à développer des modèles de pathologies humaines, à partir duquel a été isolée la lignée vdb, qui présente des défauts du système vestibulaire dus à une mutation dans le gène otog codant pour l’otogeline, et constitue un modèle de surdité chez l’homme. Le deuxième crible avait été mis en place dans le but d’approfondir nos connaissances fondamentales sur le développement du système squeletto-musculaire chez les mammifères, plus particulièrement chez l’homme et le bovin. Les analyses de deux lignées issues de ce crible, GMA24 et GMA06, ont permis d’identifier la mutation dans le gène Phex pour les souris GMA24 qui présentent un retard de croissance et constituent un modèle pour la maladie XLH (X-Linked Hypophosphatemic Rickets) chez l’homme. Pour les souris GMA06 présentant une hypermusculature, la mutation a été localisée sur le chromosome 2 et son identification est en cours. Ces trois modèles murins constituent des outils intéressants qui peuvent s’ajouter aux modèles déjà existants pour la surdité, la maladie XLH et les maladies type myopathies pour mieux appréhender les mécanismes moléculaires impliqués dans ces pathologies et les interactions génétiques mises en jeu dans l’objectif de tester de nouvelles thérapies
The random chemical mutagenesis with the Ethyl-Nitroso-Urea (ENU), whose power has been widely demonstrated during these last decades in the murine models creation, is a remarkable and essential tool in functional genomics. This approach is indeed a significant contribution to the understanding of the genes’ function and regulation; it also establishes an accelerator to identify the key elements in a signaling pathway. This systematic approach, based on the simultaneous screening of a large number of mice, requires no prior knowledge on the identity and the function of the studied genes. The mutants are identified through specific, hierarchical and non-invasive phenotypic screens. The identification of the gene and the causal mutation responsible for the mutant phenotype are achieved by gene mapping by using a series of polymorphic markers or by new generation sequencing. The objective of my thesis has been to characterize at the phenotypic and molecular level 3 independent murine lines from two ENU screens, a recessive and a sensitized dominant one. The first screen aimed to develop models for human diseases, from which has been isolated the vbd murine line, presenting vestibular system defects due to a mutation in the Otog gene coding for the otogelin. This mouse line presents a model for human deafness. The second screen had been established in order to deepen our fundamental knowledge on the skeletto-muscular system development in mammals, more particularly in humans and cattle. Analyzes of two lines from this screen, GMA24 and GMA06, have allowed to identify the mutation in Phex gene for GMA24 mice showing a growth retardation and modeling XLH (X-linked Hypophosphatemic rickets) disease in humans. For the GMA06 mice presenting an increase muscle mass, the mutation has been localized on chromosome 2 and its identification is in progress. These three murine models are interesting added tools to the existing models for deafness, XLH and myopathies diseases for a better understanding of the molecular mechanisms and genetic interactions involved in these pathologies and so testing new therapies

Thesis (Ph. D.)--University of Adelaide, Dept. of Applied Mathematics, 1994.
Copies of author's previously published articles inserted at back. Includes bibliographical references (leaves 211-220).

The aim of this work was to study the apparent asymmetry in the breast muscles in some individual broiler chickens strain "Cobb 500", with a view to revealing the cause(s) of this asymmetrical growth. Birds showing apparent asymmetry in the breast were selected to study their breast muscles anatomically and histochemically in comparison with unselected birds as controls. The skeleton was studied in these birds too. Selected chickens had heavier body and muscle weight, and higher growth rate than the controls. However, no significant differences were obtained between the two sides of the pectoralis or supracoracoideus muscles in either group of chickens, although the degree of asymmetry of the pectoralis muscle in selected chickens was higher than in the controls. The distribution of pectoralis muscle weight (degree of asymmetry) was normally distributed in both groups of chickens. The histochemical study on the pectoralis muscle revealed that there were significant differences in fibre number and diameter between the anterior (region A) and mid part (region B) of the pectoralis muscle in both groups of chickens, in that there were more FG fibres in region B, whereas region A has more FOG and SO fibre number. The diameter of fibre type in region A was significantly larger than in region B. Differences in fibre diameters were obtained between the right and left side of the pectoralis muscle. FG and SO fibres in the left anterior side of pectoralis muscle were growing significantly faster than in the right side in selected chickens, and control (against body weight or muscle weight). However, no significant differences were obtained for fibre numbers per square millimeter either between the two sides in control or in selected chickens. Many measurements on the skeleton were taken to study the shape of the sternum and the rib-cage in both groups of chickens. The essential differences were the depth of the keel, shape of the rib-cage, and the shape of the ribs. In selected chickens, depth of the keel at the right side was significantly deeper than the left, consequently the width and height of the keel in the left side were significantly greater than in the right side. As a result, the breast angle at the right side was significantly larger than the left one. Statistical analysis revealed significant differences in the shape of the ribs between the two sides. Ribs at the left side had significantly greater: arc and chord length, enclosed area and height; than the right side. In addition the orientation dorsal angle of the left ribs was significantly greater than the right. As a result, selected birds had faster bone growth, shorter, and less bone weight than the controls, in addition to the deformities in the shape of the keel, rib-cage, and ribs. From the results, it would appear that the asymmetry in the shape of the keel and rib-cage could be the consequence of the high growth rate in body weight and increased breast muscle weight, without increase of the growth of skeletal mass. Such disproportionate change in body parts could be the result of direct selection for increased amount of breast muscles. This problem could be reduced by restriction of early growth and include the concept of the skeletal growth in the selection-programme indices.

Le vieillissement de la population mondiale amène à se poser la question : comment bien vieillir ? La chute de personnes âgées est connue pour être le point de départ d’un cercle vicieux conduisant trop souvent à la perte d’autonomie avec un réel coût de prise en charge pour la société et le patient. La chute est la conséquence de la perte d’un équilibre efficace et économe en énergie, résultant d’une synergie entre le squelette, les muscles et les capacités cognitives. L’objectif de cette thèse est de rechercher les premiers signes d’un vieillissement pathologique afin de repérer, en amont, les personnes à risque. La première partie présente la caractérisation de l’alignement postural d’adultes jeunes et vieillissants, par l’analyse de reconstructions 3D du squelette obtenues à partir de radiographies bi-planaires. Elle permet d’identifier un invariant et des stratégies de compensation chez les sujets vieillissants. La deuxième partie s’intéresse au système musculaire via la reconstruction 3D de muscles à partir d’images IRM, pour des adultes jeunes et vieillissants avec déformations rachidiennes. Elle permet de mettre en évidence des changements musculaires en lien avec l’altération de la posture. La dernière partie de cette thèse aborde l’adaptation et la personnalisation d’un modèle biomécanique musculo-squelettique calculant les efforts résultants au niveau inter-vertébral. Une boucle de contrôle vise à limiter ces efforts en activant les muscles requis. L’utilisation de la géométrie 3D personnalisée dans ce modèle, ouvre à la voie à une compréhension fine des mécanismes de compensation se produisant au cours du vieillissement, normal ou pathologique. Cette partie révèle l’influence de l’altération de la posture sur le pattern de recrutement musculaire. Cette thèse met en évidence des altérations au cours du vieillissement, ce qui pourrait ouvrir la voie à l’identification de biomarqueurs permettant une meilleure prise en charge en amont des personnes vieillissantes à risque
Aging of the global population challenges scientific community in finding ways to live longer, but also in better condition. Falling of the elderly is known to be the start of a vicious cycle leading to loss of autonomy, involving great costs for the society as well as for the patient. Falling is the consequence of the loss of an efficient balance involving skeleton, muscles and cognitive capacities. The objective of this PhD was to search for early changes leading to pathological aging, in order to detect people at risk. The first part reports the characterization of the postural alignment, of both young and older asymptomatic adults, from 3D reconstruction of their skeleton based on bi-planar X-rays. An invariant was found and compensatory strategies have been identified for aging adults. The second part focuses on the 3D reconstructions of the muscles, based on MRI images, both for young adults and older adults with spinal deformities. This part identifies muscular changes in relation with postural alterations. The last part of this PhD tackles the adjustment and personalization of a biomechanical musculo-skeletal model computing the resulting load in the inter-vertebral joint. The control loop approach of the model aims to limit these loads by activating appropriate muscles. The use of 3D personalized geometry as input of the model allows a better understanding of specific compensatory mechanisms occurring during aging or pathological evolution. This part reveals the influence of the postural alteration on the muscular recruitment pattern. This PhD brings to light aging alterations of the skeleton and muscles; this could lead to biomarkers’ identification allowing a better identification of aging people at risk

L’objectif de cette thèse était le développement d'un substitut bio-hybride pour la reconstruction du continuum tendon-os sur le principe de l’ingénierie tissulaire. Après une analyse bibliographique exhaustive des structures natives et de leur environnement, nous avons d'abord proposé la réalisation de chaque système séparément en utilisant des scaffolds en polycaprolactone réalises par electrospinning. Dans un premier temps, nous avons combiné l’electrospinning et l’electrospraying pour produire un scaffold composé de polycaprolactone et d’hydroapatite avec une structure en forme de nid d'abeille. Notre hypothèse était de doter le substitut d'une structure biomimétique favorisant l'adhésion, la colonisation et la différenciation cellulaire. L'analyse mécanique et biologique in vitro réalisée avec une lignée cellulaire progénitrice et des tests organotypiques a confirmé notre approche originale. Ensuite, le matériel ensemencé avec des cellules souches de moelle osseuse a été implanté avec succès par nos collaborateurs d'Amiens dans le but de traiter un défaut maxillo-facial chez un modèle de rongeur. Parallèlement, pour la reconstruction du tendon, nous avons réalisé différents scaffolds d'electrospinning, dont la taille et l'organisation (aléatoire/alignée) des fibres varient. Dans une perspective bio-inspirée, nous avons combiné les scaffold avec l'étirement dynamique pour reproduire l'entraînement physique. Sous ces stimulations mécaniques, établies d'abord avec la même lignée cellulaire progénitrice, nous avons démontré dans une deuxième étude que les CSM s'alignaient sur l'axe d'étirement et produisaient une matrice extracellulaire, ce qui a permis de conserver les propriétés mécaniques de la matrice biohybride pendant les deux semaines de la culture. Nous avons démontré que la différenciation cellulaire vers la lignée tendineuse et osseuse a été réalisée avec succès en l'absence de tout facteur de différenciation, étant spécifiquement lié aux propriétés des matériaux et à la mécanotransduction. Par conséquent, l'étape suivante, qui consiste à assembler les deux échafaudages avec une zone de transition, devrait conduire à la reconstruction de ce continuum osseux-tendon
The objective of this thesis was the development of a biohybrid substitute for the reconstruction of the bone-tendon continuum based on tissue engineering strategies. After an exhaustive bibliographic analysis of the native structures and their environment, we first proposed the realization of each system separately using electrospun polycaprolactone scaffolds. At first, we combined electrospinning with electrospraying techniques to produce a PCL-hydroapatite scaffold with honeycomb cavities. Our hypothesis was to provide the substitute with a biomimetic structure favoring cell adhesion, spreading and differentiation. The in vitro mechanical and biological analysis performed with a progenitor cell line and withorganotypic assays confirmed our original approach. Then, the material seeded with bone marrow stem cells was successfully implanted by our collaborators in Amiens with the objective of treating a maxillofacial defect in a rodent model. In parallel, for the tendon reconstruction, we investigated several electrospinning processes, varying fibers’ size and organization (random/aligned). In a bioinspired perspective, we combined the choice of the scaffold with dynamic stretching to reproduce physical training. Under those mechanical stimulations, established first with the same progenitor cell line, we demonstrated in a second study that MSCs aligned with the stretching axis and produced extracellular matrix, which in turn allowed to keep the mechanical properties of the biohybrid scaffold all over the 2 weeks of culture. We demonstrated that cell differentiation towards tendon and bone lineage was successfully achieved in the absence of any differentiation factor, being specifically related to materials properties and mechanotransduction. Therefore, the next step consisting in the assembly of both scaffolds with a transition area should lead to this bone-tendon continuum’s reconstruction

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

Musculo – skeletal stress in nursing staff and its association with health disorders Aim of the study was to investigate health complaints and work pressure among nursing staff, and to evaluate the associations of musculo - skeletal stress with their health disorders. Methods. During October - December, 2003, 300 employees of one Kaunas hospital were interviewed. In this case - control study, 100 respondents were included into the case group, and 200 were controls. The main criterion for the selection of controls was no engagement in activities characteristic of a nurse and an assisting nurse. The questionnaire was anonymous. Statistical analysis of the obtained findings was performed using SPSS 12 statistical software package. Results. Nurses whose work was not related to lifting weights (the control group) more frequently complained of pains in the neck (87,5 %), the shoulder girdle (91,7 %), and the back (93,2 %), whereas those whose work was related to lifting weights, more frequently complained of pains around the waist (73,0 %) and in the legs (94,9 %). The study showed that the pain syndrome in the arms by two times more frequently occurred in the control group (12,1 % compared to 6,0 % in the case group), which is characteristic of the sedentary job, especially computer work. Pain syndrome in the legs by four times more frequently occurred in the case group, compared to controls, which is typical of work related to weight lifting. The evaluation of the activities... [to full text]

Introdução: A suplementação de creatina tem surgido na literatura como uma potencial estratégia terapêutica não farmacológica em diversas condições caracterizadas por disfunções musculares e baixa massa muscular, incluindo as doenças reumatológicas pediátricas. O objetivo deste estudo foi avaliar a eficácia e a segurança da suplementação de creatina em pacientes com lúpus eritematoso sistêmico de início juvenil (LESJ). Métodos: Trata-se de um estudo duplo-cego, crossover, balanceado e controlado por placebo. Os voluntários (n = 15) foram randomizados em duas condições que receberam creatina ou dextrose por 12 semanas, interpassadas por um período de washout de 8 semanas. A função muscular foi avaliada por testes de uma repetição máxima (1 RM), Timed-Up-And-Go, Timed-Stands e de preensão manual. Ainda, foram avaliados a composição corporal, os marcadores bioquímicos do remodelamento ósseo, a aptidão aeróbia, os parâmetros de qualidade de vida e a capacidade funcional dos voluntários. As possíveis alterações no consumo alimentar foram avaliadas por três recordatórios alimentares de 24h, enquanto o conteúdo de fosforilcreatina muscular foi avaliado por meio de espectroscopia de fósforo por ressonância magnética (31P-ERM). A segurança da intervenção foi avaliada por parâmetros laboratoriais e por clearance de 51Cr-EDTA e, por fim, os eventos adversos foram registrados durante todo o estudo. Resultados: Não houve diferença significativa no conteúdo intramuscular de fosforilcreatina entre as condições, antes e após as intervenções (creatina - Pré: 20,5 ± 2,6/ Pós: 20,4 ± 4,1; placebo - Pré: 19,8 ± 2,0/ Pós: 20,2 ± 3,2 mmol/kg peso úmido; p = 0,70 para interação entre condições). Ainda, provavelmente, como consequência do conteúdo intramuscular ter se mantido inalterado, não houve diferença significativa entre as condições para todos os parâmetros analisados (p > 0,05). Além do clearance de 51Cr-EDTA não ter sido alterado com a suplementação de creatina, nenhum efeito adverso foi observado. Conclusão: O protocolo de suplementação de creatina (0,1 g/kg/d) por 12 semanas foi bem tolerado e livre de efeitos adversos. Entretanto, a suplementação de creatina não foi eficaz no aumento do conteúdo intramuscular de fosforilcreatina, na melhora da função muscular, aptidão aeróbia, composição corporal e parâmetros de qualidade de vida em pacientes com LESJ
Introduction: Creatine supplementation has emerged as a promising non-pharmacological therapeutic strategy to counteract muscle dysfunction and low lean mass in a variety of conditions, including in pediatric and rheumatic diseases. The objective of this study was to examine the efficacy and safety of creatine supplementation in childhood systemic lupus erythematosus (C-SLE). Methods: C-SLE patients with mild disease activity (n=15) received placebo or creatine supplementation in a randomized fashion using a crossover, double-blind, repeated-measures design. The subjects were assessed at baseline and after 12 weeks in each arm, interspersed by a 8-week washout period. The primary outcomes was muscle function, as assessed by a battery of tests including one-maximum repetition (1-RM) tests, the Timed-Up-And-Go test, the Timed-Stands test, and the handgrip test. Secondary outcomes included body composition, biochemical markers of bone remodeling, aerobic conditioning, quality of life, and physical capacity. Possible differences in dietary intake were assessed by three 24-h dietary recalls. Muscle phosphorylcreatine content was measured through phosphorus magnetic resonance spectroscopy (31P-MRS). The safety of the intervention was assessed by laboratory parameters and kidney function was measured by the 51Cr-EDTA clearance. Additionally, self-reported adverse events were recorded throughout the trial. Results: Intramuscular phosphorylcreatine content was not significantly different between creatine and placebo before or after the intervention (creatine - Pre: 20.5 ± 2.6, Post: 20.4 ± 4.1, placebo - Pre: 19.8 ± 2.0; Post: 20.2 ± 3.2 mmol/kg wet muscle; p = 0.70 for interaction between conditions). In addition, probably as a consequence of the lack of change in intramuscular phosphorylcreatine content, there were no significant changes between placebo and creatine for any muscle function and aerobic conditioning parameters, lean mass, fat mass, bone mass, and quality of life scores (p > 0.05). The 51Cr-EDTA clearance was not altered by creatine supplementation and no side effects were noticed. Conclusion: a 12-week creatine supplementation protocol at 0.1 g/kg/d is well tolerable and free of adverse effects but did not affect intramuscular phosphorylcreatine, muscle function, free-fat mass or quality of life in C-SLE patients with mild disease activity

Hintergrund und Zielsetzung: DMD ist die häufigste Form der Muskeldystrophie im Kindesalter und bis heute unheilbar. Sie wird durch das Fehlen des Proteins Dystrophin verursacht, welches verschiedene Signaltransduktionswege beeinflusst. Das Anliegen der Arbeit ist die Untersuchung und Modulation von Signaltransduktionswegen, die als alternative Therapiestrategie den Verlust von Dystrophin kompensieren könnten. Experimentelle Strategie: Für die Charakterisierung von Dystrophin nachgeschalteten Prozessen wurden mRNA-Expressionsanalysen in Muskelgeweben von DMD-Patienten und einem DMD-Brüderpaar mit einem infrafamiliär unterschiedlichen Verlauf der DMD durchgeführt. Aus diesen Expressionsdaten wurde erstmalig ein Petri-Netz entwickelt, welches Dystrophin mit in diesem Zusammenhang bisher unbekannten Signaltransduktionswegen verknüpft. Das Petri-Netz wurde auf Netzwerkintegrität und –verhalten mittels Invarianten- (INA) und theoretischen Knockout- (Mauritius Maps) Analysen untersucht. Durch beide Methoden läßt sich der maßgebliche Teilsignalweg bestimmen. In diesem Signalweg wurden die Proteinaktivität und die Genexpression durch siRNA, Vektor-DNA und chemische Substanzen in humanen SkMCs moduliert. Anschließend wurden die Proliferation und die Vitalität der Zellen sowie auch die Expression auf mRNA- und Protein-Niveau untersucht. Ergebnisse: RAP2B und CSNK1A1 waren in dem DMD-Brüderpaar differentiell exprimiert und konnten erstmalig in einem neuen, komplexen Signalweg in Zusammenhang mit Dystrophin nachgeschalteten Prozessen dargestellt werden. Mittelpunkt dieses Signalweges ist die De- und Aktivierung des Transkriptionsfaktors NFATc. Seine Zielgene umfassen neben anderen den negativen Proliferationsfaktor p21, das Dystrophin homologe UTRN und den Differenzierungsfaktor MYF5. Folglich würde ein Anstieg von UTRN eine unerwünschte Reduktion der Proliferationsrate von Myoblasten implizieren. Letzteres konnte bereits nachgewiesen werden und stellte das Motiv für weitere Studien dar. Jedoch zeigten siRNA- und Vektor-DNA-Experimente, daß NFATc nicht der ausschlaggebende Faktor für diese Zielgene ist. Die Substanzen Deflazacort (DFZ) und Cyclosporin A (CsA) wurden dagegen beschrieben, die Aktivierung von NFATc zu beeinflussen. Die Ergebnisse zeigten, daß beide Substanzen die Proliferation von Myoblasten erhöhen können. Die gleichzeitige Applikation von DFZ und CsA führte zu einem Anstieg der UTRN-Expression. Schlußfolgerung: Die Modulation der Proliferation und UTRN-Expression ist unabhängig von einander möglich. Entsprechend der Grundidee der Arbeit zeichnet sich eine neue Therapiestrategie ab, welche Dystrophin nachgeschaltete Prozesse einbezieht.
Background and aim: DMD is the most common muscular dystrophy in childhood and incurable to date. It is caused by the absence of dystrophin, what influences several signal transduction pathways. The thesis is interested in the investigation and modulation of signal transduction pathways that may compensate the lack of dystrophin as an alternative therapy strategy. Experimental strategy: To study Dystrophin downstream pathways the mRNA expression of DMD patients and two DMD siblings with an intra-familially different course of DMD were analysed in muscle tissue. On the basis of these expression data a Petri net was first developed implicating signal transduction pathways and Dystrophin downstream cascades. Invariant (INA) and theoretical knockout (Mauritius Maps) analyses were applied for studying network integrity and behaviour. Both methods provide information about the most relevant part of the network. In this part modulation of protein activity and of gene expression using siRNA, vector-DNA, and chemical substances were performed on human SkMCs. Subsequently, the cells were studied by proliferation and vitality tests as well as expression analyses at mRNA and protein level. Results: RAP2B and CSNK1A1 were differently expressed in two DMD siblings, and first are part of a signal transduction pathway implicating Dystrophin downstream processes. The central point of this pathway is the de- and activation of the transcription factor NFATc. Its target genes are, among others, the negative proliferation factor p21, the Dystrophin homologue UTRN, and the differentiation factor MYF5. Consequently, an increase in UTRN implicates an undesirably reduced myoblast proliferation rate. Latter was found in DMD patients and was target for further studies. But, siRNA and vector DNA experiments showed that NFATc is not the decisive factor for the target genes. Deflazacort and cyclosporin A are known to influence the activation of NFATc. The results first showed that both substances do induce myoblast proliferation. The use of deflazacort in combination with cyclosporin A resulted in an increase of UTRN expression. Conclusion: The modulation of proliferation and UTRN-expression independently of each other is possible. According to the basic idea of this study, a new therapeutic strategy becomes apparent, which considers Dystrophin downstream processes.

Chemotherapy is an effective first-line cancer-treatment to slow or even cure cancer. Despite it being widely used to treat a variety of cancers, the majority of agents used induce a myriad of serious sequalae. Recently, chemotherapy emerged as a key contributing factor to the induction of devastating wasting condition, cachexia. Cachexia involves the progressive loss of body mass, underscored by severe skeletal muscle wasting and dysfunction (skeletal myopathy). Unravelling the molecular mechanisms involved in the onset and persistence of chemotherapy-induced cachexia represents a complex scientific challenge and is of great clinical interest to identify novel drug targets and efficacious adjuvants. This thesis characterised the impact of individual chemotherapeutic agents on the skeletal muscular system of mice [doxorubicin (DOX) and irinotecan (IRI), 5-fluorouracil (5FU)] and evaluated the therapeutic efficacy of mitoprotective adjuvant candidates, sodium nitrate (with DOX) and BGP-15 (for 5FU and IRI) to protect body mass and skeletal muscle during chemotherapy. Additionally, since chemotherapeutic agents are usually administered to cancer patients in combination regimens which might escalate cachexia, we also characterised the impact of the ‘7+3’ (cytarabine and daunorubicin) chemotherapy induction regimen (CIR) utilised as standard treatment against acute myeloid leukemia. In this regard, we developed and characterised a novel murine model of AML CIR-induced cachexia. We also used this model to trace the course of cachexia during and after treatment and to evaluate whether voluntary exercise could be protective. The major findings of thesis were that the onset and severity of chemotherapy-induced cachexia is agent/regimen specific. While DOX, an anthracycline and topoisomerase-II inhibitor, and IRI, a topoisomerase- I inhibitor, induced a cachectic phenotype characterised by diminished body composition indices, and skeletal myopathy, 5FU, an anti-metabolite, did not cause cachexia. Interestingly, the multi-agent CIR induced severe cachexia. The recovery post-CIR was mixed with skeletal muscle mass returning to normal levels, while body and lean mass not completely recuperating in the 2-week recovery period. At the molecular level, the expression of key structural cytoskeletal proteins, i.e. dystrophin, were impacted by IRI and 5FU whether skeletal myopathy was observed or not. These data suggest that loss of dystrophin might be an early event in the myopathy associated with cachexia. With regard to the adjuvant candidates evaluated, sodium nitrate was not protective against DOX-induced cachexia, despite protecting against early signs of cardiomyopathy. BGP-15 displayed modest protection against IRI-induced cachexia but was not afforded the opportunity when evaluated in combination with 5FU. Alongside the CIR voluntary activity was not protective against cachexia, rather it potentiated CIR-induced cachexia, likely driven through enhanced loss of fat mass. Overall, these findings highlight that further investigation is required regarding the efficacy of mitoprotective adjuvant therapies against chemotherapy-induced cachexia.

Copies of author's previously published articles inserted at back
Bibliography: leaves 211-220
viii, 220 leaves : ill ; 30 cm.
Summary: Investigates various problems relating to scoliosis and manual load handling. Introduces the idea of deducing optimal load lifting techniques which take into into account multiple joint/muscle complexes
Thesis (Ph.D.)--University of Adelaide, Dept. of Applied Mathematics, 1994

Title: Musculoskeletal assessment analysis of triathletes Objectives: The main aim of this master degree thesis is the diagnostics of the current condition of the complex kinesiological examination of musculo-skeletal system in elite performance triathtletes of the Czech National Team level (Junior and U23 categories), including compensational exercises specifically devised for triathlets. Futhermore with the help of the selected data this study is expected to draw some basic conclusions and establish the typical postural characteristics of athletes related to the sport of triathlon. Methods: In this thesis we have used following testing methods: complex musculo skeletal examintation including evaluation of medical history, an antropometrical screening, standing static postural assessment, gait assessment, soft-tissue palpation exam, assessment of the fundamental movement patterns of an individua, examination of neurological reflexes, flexibility and range of motion screening. Based on the above mentioned tests we have received a complete functional and musculo skeletal system profile of each triathlete's. For collection we have used the SPSS 22 data analysis software. For the gender diferentiation we have used non-pairred T-Test and for perofrmance variability within the 3 levels we have used...