Introdução: A escolha de parâmetros mais eficientes na aplicação terapêutica de um recurso de eletroterapia garante uma efetividade clínica maior ao tratamento realizado. O modo contínuo de aplicação do ultrassom vem sendo pouco estudado nas fases agudas das lesões. Objetivo: Avaliar os efeitos do ultrassom contínuo nas fases inflamatória, proliferativa e de remodelação no processo de regeneração do tecido conjuntivo muscular de ratos. Material e Métodos: Trinta ratos foram submetidos à lesão muscular nos músculos tibial anterior de ambos os membros traseiros e divididos em dois grupos, sendo o grupo Lesado (L) composto pelos membros traseiros esquerdos dos animais e o grupo lesado tratado com ultrassom (LUS), composto pelos membros traseiros direitos. Cada grupo foi subdividido em seis pequenos grupos (n = 5), que foram formados de acordo com o número de dias após a lesão em que os animais foram mortos, sendo no 3º, 7º, 14º, 21º, 28º e 35º dias. Após 24 horas da realização da lesão cirúrgica, o Grupo LUS começou a receber a aplicação do ultrassom no modo contínuo, área de radiação efetiva de 1cm2, com freqüência de transdutor de 1MHz, intensidade de 0,2W/cm2, por 1 minuto, uma vez ao dia. Os membros traseiros esquerdos submetidos à lesão cirúrgica e não tratados serviram como grupo controle (L). Para análise morfológica das fibras musculares foram realizados cortes seriados de 10 ?m de espessura, obtidos através de em um micrótomo criostato. Os músculos foram congelados em uma posição onde os cortes pudessem fornecer imagens longitudinais das fibras musculares e da fenda incisional. Cada série de corte foi obtida a partir da região superficial do músculo, a cada 100 ?m de profundidade e forneceu duas lâminas que foram coradas com a técnica Hematoxilina Eosina. A contagem de células foi feita em uma área de 0,625mm2 (área de um campo microscópico), com auxílio de uma câmera acoplada ao microscópio e monitor de vídeo e tendo como ponto de referência os limites da fenda incisional, ou seja, a região com fibras musculares lesionadas de um lado e as células infiltradas na fenda incisional de outro. A determinação da densidade de leucócitos polimorfonucleares e mononucleares, e de fibroblastos foi feita através da contagem média de 10 campos por amostra, sendo cinco campos representativos do tecido muscular seccionado e cinco campos da fenda incisional, totalizando 300 campos por grupo experimental (30 ratos ao todo). As células com morfologia de células satélites ou mioblastos foram ignoradas. Resultados: A análise da contagem de células revelou diferenças significativas (p <= 0,05) entre o número de células polimorfonucleares no 3º, 7º, 21º e 28º dias de tratamento com as seguintes xiii médias: L3 = 27,0; L7 = 29,6; L21 = 20,0 e L28 = 12,8 e LUS3 = 43,2; LUS7 = 29,2; LUS21 = 8,8 e LUS28 = 3,6. O número de fibroblastos foi significativamente maior (p<=0,05) apenas no 21º dia com L21 = 14,2 e LUS21 = 27,0. Conclusão: O ultrassom contínuo promoveu um aumento na densidade de células polimorfonucleares e fibroblastos durante o processo de regeneração do músculo tibial anterior de ratos, incrementando a recuperação deste tecido. / Introduction: The choice of more efficient parameters in the ultrasound application allows a major clinical effectivity to the performed treatment. The therapeutic continuous ultrasound has been being poorly studied in injury acute phases. Objective: To assess the effects of the continuous ultrasound in the inflammatory, proliferative and remodeling phases of rats\' conjunctive muscular tissue regeneration process. Material and Methods: Thirty rats were submitted to muscular lesion in tibialis anterior muscle of both rear limbs. They were divided into two groups, the lesion group (L) consisting of the rats left rear limbs, and the lesion group treated with ultrasound (LUS) consisting of the right rear limbs. Each group was subdivided into six small groups (n = 5), which were formed according to period of time (in days) after the lesion in which the rats were killed, on the 3rd, 7th, 14th, 21st, 28th and 35th days. Twenty four hours after performing the surgical lesion, the LUS group received the first 1MHz continuous ultrasound treatment with effective radiating area = 1cm2, intensity of 0.2W/cm2, for one minute, once a day. The left rear limbs submitted to the surgical lesion and not treated at all composed the control group (L). To perform the muscle fibers morphological analysis, a series of 10?m thick slices were made using a microtome with criostate. The muscles were frozen in a position which the slices could provide longitudinal images of the muscle fibers and incision gap. A series of samples were collected from the lesion area surface, and further two deeper series, at 100?m distance from one another. Two slices were thus provided and stained with hematoxilin-eosin. Cell count was made on an area of 0.625 mm2, by means of a camera coupled to the microscope and to a video monitor. The reference point were the limits of the incision gap, which is a region with injured muscle fibers in one side and infiltrated cells of the incision gap on the other. The determination of the density of polymorphonuclear and mononuclear leukocytes, and fibroblasts was made by the exam of 10 fields per sample (five fields representing the cut muscular tissue and more five fields of the incision gap), totaling 300 fields per group. Cells with myoblast or satellite cells morphology were ignored. Results: The cells counting analysis revealed significant differences (p <= 0.05) in the density of polymorphonuclear cells on the 3rd, 7th, 21st and 28th treatment days and the means are: L3 = 27; L7 = 29.6; L21 = 20; L28 = 12.8 and LUS3 = 43.2; LUS7 = 29.2; LUS21 = 8.8; LUS28 = 3.6. The density of fibroblasts was significant greater (p <= 0.05) only in the 21st day with means of L21 = 14.2 and LUS21 = 27. Conclusion: The continuous ultrasound increased the density of polymorphonuclear and fibroblasts cells during the anterior tibialis muscle regenerating process of rats, enhancing the recovery of this tissue.

A proposta deste estudo foi verificar o efeito da sonoforese com Arnica montana sobre a fase inflamatória aguda de uma lesão muscular. Para isso, 40 ratos Wistar machos (300±50 g), lesados cirurgicamente, foram divididos em 4 grupos: grupo controle (C), 10 ratos lesados, não submetidos a tratamento algum; grupo ultra-som (US), 10 ratos lesados, tratados com US; grupo ultrasom+ arnica (US+A), 10 ratos lesados, tratados com sonoforese com gel de arnica; e grupo arnica (A), 10 ratos lesados, tratados com massagem de gel de arnica. O ultra-som com freqüência de 1 MHz e 0,5 W/cm2 de intensidade foi aplicado 24 h após a lesão uma vez ao dia durante 3 minutos, por 3 dias; o grupo US+A recebeu aplicação de ultra-som idêntica, mas com gel de arnica; o grupo A recebeu massagem de gel de arnica nos mesmos tempo e período. O terço médio do músculo tibial anterior lesado foi removido, montando-se séries de lâminas para análise quantitativa (contagem de células mononucleares e polimorfonucleares) e qualitativa (morfologia das fibras musculares). Os resultados foram tratados estatisticamente, com nível de significância p<0,05. Foi encontrada maior densidade de células mononucleares nos grupos US, US+A e A, sem diferença entre estes, mas com diferença significativa (p<0,0001) destes para o grupo controle, onde a densidade de células polimorfonucleares também foi significativamente diferente (p=0,0134) dos demais. Ainda quanto a essa densidade, não houve diferença entre os grupos US e US+A, mas houve entre o grupo A (p=0,0134) e os demais. A análise qualitativa revelou estágio mais avançado de regeneração nos tecidos dos grupos US e US+A igualmente. A similaridade de resultados entre os grupos US e US+A leva a concluir que a sonoforese com arnica não foi mais eficaz que o ultra-som na recuperação da lesão muscular. Sugere-se que o US pode ter anulado ou minimizado o efeito regenerativo da Arnica montana por sonoforese na fase inflamatória aguda da lesão muscular. / This study aimed at verifying the effects of ultrasound associated to Arnica montana (phonophoresis) onto the acute phase of an inflammatory muscle lesion. Forty Wistar male rats (300±50 g), of which the Tibialis Anterior muscle was surgically lesioned, were divided into 4 groups (n=10 each): control group received no treatment; the ultrasound group (US) was treated with 1-MHz, 0.5W/cm2-intensity US during 3 minutes, once a day, for 3 days; the US+A group was treated with arnica phonophoresis (the same US parameters plus arnica gel); and the arnica group (A) was submitted to massage with arnica gel, also during 3 minutes, once a day, for 3 days. Treatment started 24 h after the surgical lesion. On the 4th day after lesion animals were sacrificed and sections of the lesioned, inflamed muscle were removed for quantitative (mononuclear and polymorphonuclear cell count) and qualitative analysis (muscle fiber morphology). Collected data from the 4 groups were statistically analysed and the significance level set at p<0.05. Results show higher mononuclear cell density in all three treated groups with no significant difference between them, but values were significantly different (p<.0001) when compared to control group\'s. As to polymorphonuclear cell density, significant differences were found between control group (p=0.0134) and the other three groups; the arnica group presented lesser density of polymorphonuclear cells when compared (p=0.0134) to the other groups, and no significant differences were found between US and US+A groups. Qualitative analysis showed a more advanced regeneration stage in tissues from both US and US+A groups. The similarity of results between US and US+A groups points to ineffectiveness of Arnica montana phonophoresis, US having seemingly checked or minimized its anti-inflammatory effect.

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

A maioria dos estudos explora o enriquecimento ambiental (EA) como uma forma de estudar o comportamento do animal, função cerebral e memória. Além disso, o EA pode estimular a atividade física voluntária. No entanto, pouco se sabe sobre o impacto deste estímulo sobre o perfil de estresse oxidativo no coração, assim como nos parâmetros cardiometabólicos. O objetivo do presente estudo é verificar os efeitos da atividade física voluntária sobre os parâmetros cardiometabólicos e sobre o perfil de estresse oxidativo no coração e músculo sóleo de ratos jovens e velhos. Foram utilizados 32 ratos, machos, Wistar, separados em 4 grupos: jovens controle (JC), jovens com enriquecimento ambiental (JEA), velhos controle (VC), velhos com enriquecimento ambiental (VEA). Os animais foram mantidos em seus grupos por 6 semanas, sendo que após esse período todos os animais foram avaliados (teste de esforço, peso, glicemia). Os resultados demonstraram alterações na composição corporal: aumento da gordura marrom no grupo VEA comparado ao grupo VC e redução da gordura periepididimal no grupo VEA comparado ao grupo VC. Houve também uma melhora no desempenho físico (tempo de corrida e velocidade) observado no teste de esforço do grupo VE comparado ao VC. A enzima citrato sintase é menor no VC comparado ao JC, sendo que o EA faz desaparecer essa diferença, deixando o grupo VEA semelhante ao JEA. Em relação ao perfil de estresse oxidativo, houve redução dos pró-oxidantes no grupo JEA tanto no coração quanto no músculo sóleo; já no grupo VEA houve aumento da enzima antioxidante superóxido dismutase (SOD) e foi possível observar uma tendência de aumento na glutationa peroxidase (Gpx). O EA é capaz de alterar a composição corporal, melhorar a performance e aumentar a SOD no grupo de ratos velhos. No grupo de ratos jovens reduz o dano oxidativo tanto no músculo sóleo quanto no coração, mas não provoca adaptações relevantes no sistema de defesa antioxidante enzimático / Most studies exploring environmental enrichment (EE) as a way to study the animal\'s behavior, brain function and memory. In addition the EE can stimulate voluntary physical activity however, little is known about the impact of stimulus Profile oxidative stress in the heart, as in cardiometabolic parameters. The aim of this study was to access the effects of voluntary physical activity (EE) on cardiometabolic parameters and the oxidative stress profile in soleus muscle and heart of young and old male rats. Thirty-two male Wistar rats were randomly assigned into four groups: young control (YC), young environmental enrichment (YE), old control (OC), old environmental enrichment (OE). After six weeks, the weight, stress test and blood glucose were evaluated in all groups. The data showed body composition changes: increased in the brown fat in OE group (0.072 ± 0.032) compared to OC group (0.051 ± 0.012); and reduced fat periepydidimal in OEA group (1.25 ± 0.48) compared to OC group (1.87 ± 0.63). We also observed a physical performance improvement (running time and speed) in the stress test of OE group (232.14 ± 102.50) compared to OC (51.67 ± 44.74). The enzyme citrate synthase was lower in OC compared to YC ( P= 0.0020), indicating a deleterious effect of aging,. EE has removed this difference, leaving the OE group similar to YE (P= 0.1529). Regarding the oxidative stress profile, there was a reduction of pro-oxidants in the heart (H202, carbonyls, lipoperoxidation) and in the soleus muscle (carbonyls and lipoperoxidation) of YE group. In the OE there was a significant increase of the antioxidant superoxide dismutase enzyme (SOD) and we observed a increase trend in glutathione peroxidase (GPx). EE was capable of altering body composition (brown and periepydidimal fat), improved physical performance and increased SOD in old mice group. In young group, oxidative damage reduced both in soleus muscle as in the heart, but does not caused significant adjustments in the enzymatic antioxidant defense system

Cell adhesion is essential to cell motility and tissue integrity and is regulated by the Integrin family of transmembrane receptors. Integrin binds to ligand extracellularly and provide anchor to the intracellular cytoskeleton via adhesion scaffolding proteins. In order to link cell to the surrounding matrix Integrin needs to be activated. Intracellular activation signals induce perturbations in Integrin cytoplasmic domain that are translated into a conformational change in extracellular region for high affinity ligand binding. Integrin engagement by matrix, in turn, triggers the assembly of adhesion complexes. Such early adhesions promote cytoskeletal organization with subsequent contractile activity that exerts forces against initial Integrin-matrix adhesions. In response to force, Integrin strengthens the interaction with matrix through its clustering and successive recruitment of additional adhesion components. These bidirectional regulatory loops mediated by such interactions are largely dependent on the unique function of Integrin adhesion components. / We demonstrate a novel role for the PDZ/LIM domain protein Zasp as a core component of Integrin adhesions. Specifically, Zasp colocalizes with Integrins at focal adhesion in cultured cells and myotendinous junctions in Drosophila embryos. In both cases elimination of Zasp modifies Integrin function causing consequently defects in cell spreading and muscle attachment. Zasp supports Integrin adhesion to the extracellular matrix that is required to withstand tensile forces exerted during cell spreading and muscle contraction. Furthermore, we found that the distribution of Zasp in muscle Z-lines is essential to orchestrate the cross-linking of alpha-Actinin and Actin filaments. Disruption of Zasp leads to loss of muscle cytoarchitecture, pointing to a larger role for Zasp in sarcomere assembly. Finally, we demonstrate that Zasp, in addition to alpha-Actinin, physically interacts with the Integrin- and Actin-bound cytoskeletal protein Talin. / Collectively, our results point to a dual role for Zasp as a structural scaffold. First it regulates Integrin adhesion to the extracellular matrix by interacting with the head domain of Talin at the myotendinous junctions. Second, Zasp controls sarcomere assembly by tethering the presarcomeric alpha-Actinin component to the tail domain of Talin. Zasp finding as a crucial adhesion component provides further insights on the mechanism underlying Integrin-mediated adhesion.

The fast-skeletal-muscle-fiber-specific expression of the troponin I(fast) (TnIfast) gene is driven by an Intronic Regulatory Element (IRE) located within the first intron of the gene. The IRE is a 148 bp transcriptional enhancer that contains several known and suspected cis-regulatory elements. These include the E-box, the closely-spaced MEF2 site and CACT box, the CACC site, and the CAGG element. Previous loss-of-function studies performed using the quail TnIfast IRE suggest that its activity depended on the MEF2 and CACT elements. The goal of my thesis research was to determine whether the MEF2 and CACT sites were not only necessary, but also sufficient, to support IRE activity. I prepared head-to-tail multimers of a 27-bp IRE segment that consisted largely of the near-adjacent MEF2 and CACT elements and did not contain any other known/suspected elements. These multimers were cloned upstream of a reporter gene consisting of the minimal promoter of the quail TnIfast gene linked to sequences encoding human placental alkaline phosphatase. The transcriptional capabilities of the constructs were assessed by gene transfer into the mouse soleus muscle in vivo by intramuscular injection/electroporation, and histochemical analysis of reporter enzyme plap expression including quantitative microdensitometry. I found that expression of these constructs was readily detectable and that it was markedly reduced by prior mutation of the CACT and, especially, of the MEF2 sites. These data indicate that the short DNA segment containing MEF2 and CACT elements is sufficient to drive expression in skeletal muscle and confirms the functional importance of these specific elements. / Although constructs containing the wild-type IRE 27-bp region were expressed, there was little preferential expression in fast fibers, in contrast to expression driven by the complete 148-bp IRE. Thus my results indicate that the MEF2 and CACT elements are not sufficient to drive fast fiber-type-specific expression, and suggest that additional elements outside of the 27-bp region tested are also necessary for fiber-type-specificity.

Lic. (sammanfattning) Göteborg : Univ., 2004. / Härtill 2 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 6 uppsatser.

Thesis (M.S.)--University of Missouri--Columbia, 2004. / Typescript. Vita. Includes bibliographical references (leaves 70-75). Also issued on the Internet.

Thesis (M.S.)--University of Tennessee Health Science Center, 2006. / The electronic version of this thesis is available at http://etd.utmem.edu/WORLD-ACCESS/2006-002-yan.pdf Includes bibliographical references (leaves 128-136).