Painful conditions having the origin within the musculoskeletal system is a common cause for people to seek medical care. Between 20-40% of all visits to the primal care in Sweden are coupled to pain from the musculoskeletal system. Muscle pain and impaired muscle function can be caused by muscles being repetitively overused and/or via heavy load. Skeletal muscle is a dynamic tissue which can undergo changes in order to fulfill what is best for optimal function. However, if the load is too heavy, morphological changes including necrosis, as well as pain can occur. The extension of the skeletal muscle is the tendon. Tendinopathy refers to illness and pain of the tendon. The peritendinous tissue is of importance in the features related to tendon pain. Common tendons/origins being afflicted by tendinopathy/pain are the Achilles tendon and the extensor origin at the elbow region.    Tumor necrosis factor alpha (TNF-alpha) is a cytokine that is involved in several biological processes. It is well-known for its involvement in the immune system and is an important target for inflammatory disorders such as rheumatoid arthritis. It is not known to what extent the TNF-alpha system is involved in the process of muscle inflammation and damage due to overuse.    Studies were conducted on rabbit and human tissue, tissues that either had undergone an excessive loading activity or tissue that was removed with surgery due to painful conditions. The tissues were evaluated via staining for morphology, in situ hybridization and immunofluorescence.    Unilateral experimental overuse of rabbit muscle (soleus muscle) led to morphological changes in the soleus muscle tissue bilaterally. The longer the experiment extended, the more was the tissue affected. This included infiltration of white blood cells in the tissue (myositis) and abnormal muscle fiber appearances. TNF-alpha mRNA was seen in white blood cells, in muscle fibers interpreted to be in a reparative stage and in white blood cells that had infiltrated into necrotic muscle fibers.  There was an upregulation in expressions of TNF receptor type 1 (TNFR1) and TNF receptor type 2 (TNFR2) in muscles that were markedly overused, with expressions in white blood cells, fibroblasts, blood vessel walls and muscle fibers. Immunoreactions for the receptors were seen in nerve fascicles of markedly overused muscles but only occasionally in normal muscles. The upregulations were seen for both experimental and contralateral sides. Overall the two receptors showed somewhat different expression patterns. Tendinopathy is associated with an increase in blood flow and infiltration of white blood cells in the tissue adjacent to the tendon. It is called the peritendinous tissue and is also richly innervated. The white blood cells and the blood vessels walls in this tissue were showing immunoreaction for TNFR1 and TNFR2. Two types of nerve fascicles were found in this tissue, one normally appearing when staining for nerve markers and one type with signs of axonal loss. The latter had clearly strong immunoreactions for TNFR1 and TNFR2.    The findings suggest that the TNF-alpha system is involved in both myopathies occurring due to overuse and in features in the peritendinous tissue in the tendinopathy situation. TNF-alpha and its receptors seem to be involved in degeneration but also in regeneration and healing of the tissue. The findings also suggest that TNF-alpha has effects on nerves showing axonal loss. The changes in the TNF-alpha system were seen both on the experimental side and contralaterally. / Smärta och funktionsbortfall från rörelseapparaten är vanligt förekommande. Mellan 20-40% av alla besök i primärvården är kopplade till smärta från rörelseapparaten. Det är också en vanlig orsak till sjukfrånvaro. Överansträngning inklusive repetitivt enformigt muskelarbete kan leda till muskelsmärta och bristande muskelfunktion (ex nedsatt styrka och uthållighet, inskränkt rörlighet). Muskelvävnad är en dynamisk vävnad som kan ändras utefter vilka påfrestningar den utsätts för och därigenom vilka behov den ställs inför. Men om belastningen blir för hård, alternativt återhämtningen blir för kort, kan negativa förändringar i vävnadsstrukturen uppstå, inklusive celldöd och vävnadsskada. Förlängningen av muskeln är senan. Senan är den vävnad som förbinder muskeln med skelettet. Tendinopati innefattar smärtsamma sjukdomstillstånd i senan. När sjukdom i en sena uppstår, exempelvis en smärtande hälsena, har man sett att den lösa bindväven som omger senan är av betydelse. Den genomgår morfologiska förändringar och man tror att det är den som är med och bidrar till smärtan vid tillståndet. Akillessenan och ”tennis-armbåge” är vanliga ställen för tendinopati. Akillessenan förbinder den trehövdade vadmuskeln med hälbenet. Tennis-armbåge omfattar ett område för flera musklers ursprung vid armbågen. Dessa muskler ansvarar framför allt för att sträcka i handleden. TNF-alfa är en signalsubstans som är involverad i flertalet biologiska processer. Den är känd för sin del i immunförsvaret och den är ett viktigt mål för behandling av autoimmuna sjukdomar som exempelvis reumatoid artrit. Det är inte känt om TNF-alfa är inblandad i processen som uppstår vid muskelinflammation/muskelskada efter kraftig överansträngning. TNF-alfa har flera receptorer, i det här arbetet har utbredning av TNFR1 och TNFR2 analyserats. Studier har utförts på djur (kaniner) och människa. Kaniner har genomgått ett träningsexperiment, där de utsatts för repetitiva muskelkontraktioner som lett till överansträngningsskador och muskelinflammation. Den muskel som studerats är soleus-muskeln, en del i den trehövdade vadmuskeln. Vävnadsprover har tagits från patienter med smärta i Akillessenan eller tennisarmbåge. Vävnadsproverna från kanin och människa har analyserats med färgningar för morfologi, immunohistokemi för detektering av TNF-alfa och dess receptorer samt för in situ hybridisering för detektion av mRNA i TNF-alfa systemet. Parallellt med färgningar för faktorerna i TNF-alfa systemet har uttryck för andra faktorer studerats. Ensidig överbelastning hos kaniner ledde till samma morfologiska förändringar på båda sidor, det vill säga även i muskeln i det ben som inte hade genomgått träningsexperimentet. Ju längre experimentet pågick, desto större blev de morfologiska förändringarna. TNF-alfa sågs i vita blodkroppar, TNF-alfa mRNA sågs även i förändrade muskelfibrer. Resultatet av parallella dubbelfärgningar tolkades som att dessa muskelfibrer antingen var i en regenererande process eller i en destruktiv process. TNFR1 och TNFR2 uttrycktes i större utsträckning ju längre experimentet pågick och ju mer muskelvävnaden var påverkad av inflammation. TNF receptorer sågs i vita blodkroppar, fibroblaster, muskelfibrer och nervstrukturer hos experimentdjuren. Det såg lika ut på båda sidor, inklusive det ben som inte ingått i experimentet. De två receptorerna skilde sig åt i uttryck. Vävnad från patienter med smärtande senor/smärta vid muskelursprungs-region genomgick också färgningar för faktorer i TNF-alfa systemet. Man kunde se att den lösa bindväven runt senan (den peritendinösa vävnaden) innehöll mycket blodkärl och nerver. De nerver som sågs i denna vävnad var av två typer, en som såg normal ut och en typ som uppvisade tecken på förlust av axoner. Den senare varianten hade en tydlig uppreglering av båda TNF receptorerna. Dessa resultat tyder på att TNF-alfa systemet är involverat i muskelsjukdomar som rör muskelinflammation till följd av kraftig överansträngning och i processerna i bindväven vid smärtande senor. TNF-alfa och dess receptorer verkar vara inblandade i både nedbrytning och uppbyggnad av muskelvävnad, samt påverka nerver som visar tecken på förlust av axoner. Förändringarna i TNF-alfa systemet sågs både på experimentsidan och kontralateralt.

The age-related loss of skeletal muscle mass and function is accompanied by a decline in regenerative capacity. The processes that facilitate healthy muscle repair are complex, involving several phases of degradation and rebuilding of muscle tissue and the surrounding microenvironment. Specifically, myogenic progenitor cells known as satellite cells are the most influential in repairing damaged muscle tissue. Following injury, satellite cells become activated and migrate, proliferate and fuse with mature skeletal muscle fibers to restore homeostasis to the tissue. However, satellite cells do not act in isolation, a robust inflammatory response is necessary to facilitate successful and rapid healing. Macrophages are one of the first and most abundant immune cells to infiltrate damaged skeletal muscle tissue. Primarily, macrophages adapt to a proinflammatory state to clear the area of cellular debris, promote degradation of the extracellular matrix and stimulate satellite cell activation and proliferation. Afterwards, a timely transition to an anti-inflammatory state directs rebuilding of the extracellular matrix and terminal differentiation of satellite cells. Indeed, the inhibition of macrophage activity leads to impaired healing and loss of skeletal muscle function. Little is known regarding the behavior of macrophages in aged skeletal muscle following injury in humans. Thus, the objective of this dissertation is to investigate the age-related response of macrophages in human skeletal muscle, and their role in muscle repair.

Les lésions musculaires sont des événements fréquents. Le diagnostic repose sur la mesure de biomarqueurs sanguins. Les outils actuels présentent des limites qui justifient la recherche de nouveaux candidats biomarqueurs. Récemment, des petits ARNs non codants, les microARNs (miARNs), ont été identifiés. Détectables dans le plasma, certains sont spécifiques d’un tissu et ont été proposés comme de potentiels biomarqueurs de lésions tissulaires. Toutefois, leur intérêt dans le diagnostic des lésions musculaires chez l’individu sain n’est pas connu. Le but de ce travail était d’identifier et de caractériser la réponse des miARNs circulants à des lésions musculaires chez le rat.Nous avons premièrement étudié les profils plasmatiques des miARNs en réponse à des lésions musculaires myotoxiques chez le rat sain pour identifier des candidats biomarqueurs et leurs cinétiques de détection. Un criblage par RT-qPCR nous a conduits à identifier l’augmentation importante des niveaux plasmatiques de miARNs spécifiques du tissu musculaire, miR-1-3p, -133a-3p, -133b-3p, -206-3p, -208b-3p et -499-5p avec un pic de détection à 12 h. De plus, deux miARNs non spécifiques du muscle, miR-378a-3p et miR-434-3p, avaient des profils comparables. L’évaluation des performances diagnostiques a montré que les miARNs sélectionnés pouvaient discriminer les rats lésés des rats non lésés avec peu d’erreurs et une approche combinatoire nous permettait d’améliorer encore ces performances. Ces résultats ont été confirmés chez des rates femelles et des rats mâles âgés. Par ailleurs, nous avons évalué la robustesse des miARNs que nous avons sélectionnés. Malgré des profils d’expression différents des miARNs dans les fibres lentes ou rapides, le phénotype du muscle lésé avait une influence limitée sur la réponse des miARNs. Nous avons ensuite observé que la lésion d’une masse musculaire croissante ne s’accompagnait pas d’une réponse proportionnelle des miARNs circulants. Les miARNs sélectionnés n’augmentaient pas en réponse à des lésions musculaires traumatiques. Néanmoins, nous avons observé que miR-133a-3p et -133b-3p pourraient être des marqueurs intéressant pour détecter un remodelage musculaire précoce après des lésions neurologiques. Enfin, l’hémolyse et la contamination plaquettaire, deux paramètres préanalytiques connus pour induire des modifications des profils circulants, n’avaient pas d’effet sur les miARNs que nous avions identifiés.Pris dans leur ensemble, nos résultats montrent que les miARNs circulants spécifiques du muscle ainsi que miR-378a-3p et miR-434-3p, sont des biomarqueurs robuste et prometteur de lésions musculaires aigues chez le rat. / Skeletal muscle damage is an often-occuring event. Diagnosis is based on blood biomarkers assessment. Yet, the markers currently available suffer limitations and new biomarker candidates are needed. Recently, small non-coding RNA, microRNAs (miRNAs), were identified. Detectable in plasma, some miRNAs are tissue-specific and have been proposed as biomarkers of tissue damage. However, their relevance as biomarkers of skeletal muscle damage in healthy individuals is unknown. The aim of this work was to identify and characterize the circulating miRNAs response to muscle damage in rats.First, we studied circulating miRNAs response to myotoxic muscle damage in healthy rats in order to identify biomarker candidates and their detection kinetics. RT-qPCR profiling led to the identification of muscle-specific miRNAs that subtantially increased in plasma in response to muscle damage, namely miR-1-3p, -133a-3p, -133b-3p, -206-3p, -208b-3p, and -499-5p with a peak value at 12 h. Two non-muscle-specific miRNAs, miR-378a-3p and miR-434-3p, had similar profiles. The evaluation of the diagnostic accuracy has shown that selected miRNAs were able to discriminate damaged from non-damaged rats with almost no error and a combinatory approach was able to further increase this accuracy. Similar results were found in female and aged rats. Moreover, we sought to evaluate the robustness of selected miRNAs. Despite diferente expression of selected miRNAs in slow and fast fibers, the phenotype of injured muscle had a very limited influence on the plasma miRNA response. Then, we induced muscle damage in an increasing muscle mass and we observed that damage responsive miRNA response was not proportional to the extent of muscle damage. Selected miRNAs did not increased in response to traumatic muscle damage. However, we observed that miR-133a-3p et -133b-3p could be useful markers to detect an early muscle remodeling following neurologic damage. Finally, hemolysis and platelet contamination, two pre-analytical factors known to affect circulating miRNA profiles, had no effect on the miRNAs we selected.Taken together, our results show that circulating muscle-specific miRNAs as well as miR-378a-3p and miR-434-3p, are robust and promising biomarkers of acute muscle damage in rats.

<p>Obesity is associated with an increase in low grade systemic inflammation. Skeletal muscle of individuals with obesity undergo numerous biochemical and morphological alterations including an increase in ectopic lipid accumulation in skeletal muscle and increased macrophage infiltration. Increased intermuscular adipose tissue and macrophages contribute to skeletal muscle inflammation and insulin resistance by secreting elevated proinflammatory cytokines and lipids. This also contributes to reduction in skeletal muscle quality, increasing the susceptibility of muscle to damage and impairing the regenerative response to muscle. Exercise training can reduce inflammation and improve skeletal muscle quality. Importantly reductions in inflammation occur without change in adiposity. Peroxisome proliferator activated receptor g coactivator 1-a (PGC-1a) exerts protective effects on skeletal muscle against damaging insults and may improve muscle regeneration.</p><p> The primary aim of my dissertation was to determine the mechanisms that lead to deficits in skeletal muscle integrity and regeneration in persons with obesity. In Chapter 1, an introduction to the various physiological, pathological, and clinical topics is provided. In Chapter 2, we investigated how exercise training and obesity independently alter skeletal muscle extracellular vesicle (EV) miRNA (miR) content. We found that obesity alters EV miR content indicative of altered anabolic signaling, while exercise training altered EV miR content in a manner indicative of reduced inflammation. In Chapter 3, we report that overexpression of PGC-1a reduces cardiotoxin induced damage of primary human myotubes but limits the ability of undifferentiated cells to reenter the cell cycle and produce progeny that could aid in the restoration of myotubes. In Chapter 4, we demonstrate that exposure to an obesogenic environment increases cardiotoxin induced damage of primary human myotubes from obese donors. In this study we also found that the restoration of myotube fusion index was reduced in lean and obese subjects when incubated with obesogenic media. In Chapter 5 is a review and summary of the outcomes described in Chapters 2-4, a discussion of the limitations of these experiments, and a discussion of future directions.</p>

In the past decade there has been a significant increase in focus on the effect upper body vibration (UBV) has on the recovery of skeletal muscle after exercise-induced muscle damage. Recovery can be defined and investigated using a wide variety of methods. This study used three different measurements to track muscle recovery over 7 days following an exercise muscle damage protocol and applied vibration to a mathematical model. A visual analog scale (VAS) was used to measure muscle pain, a strain gauge was used to obtain maximum voluntary isometric contraction (MVIC) strength measurements, and shear wave elastography (SWE) represented muscle stiffness over the 7-day experiment. Thirty-three participants were divided into three groups. The first was a control group (C) that experienced no exercise and no therapy. The no vibration group (NV) performed the damage an exercise protocol but received no therapy. The vibration group (V) performed the same exercise protocol but also received vibration therapy. The exercise protocol consisted of 100 dumbbell curls at starting at 50% of their MVIC with one minute of rest after each set of ten. The data provided convincing evidence (27.2%, p < 0.0001) that group NV was not back to its normal stiffness after 7 days unlike group V, which was shown not to be any different from its baseline at the end of the week (9.15%, p = 0.137). Three vibration factors (����1, ����2, ����3) were added to a skeletal muscle regeneration model (SK) to simulate how vibration affects muscle regeneration. The three factors were determined by analyzing previous research to understand how vibration affects cells in the regeneration process. Adding these into SK decreased the time to recovery from about 13 days to about 7 days. Recovery was defined by reaching 10% of the original number of myofibers within the damaged muscle.

Submitted by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2018-09-27T17:53:00Z No. of bitstreams: 1 ALVARO SOSA MACHADO.pdf: 1403839 bytes, checksum: ecb430aded30475dee48fa2ac550cd0d (MD5) / Approved for entry into archive by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2018-09-27T17:53:12Z (GMT) No. of bitstreams: 1 ALVARO SOSA MACHADO.pdf: 1403839 bytes, checksum: ecb430aded30475dee48fa2ac550cd0d (MD5) / Made available in DSpace on 2018-09-27T17:53:12Z (GMT). No. of bitstreams: 1 ALVARO SOSA MACHADO.pdf: 1403839 bytes, checksum: ecb430aded30475dee48fa2ac550cd0d (MD5) Previous issue date: 2018-02-23 / A prática regular de exercício intenso é benéfica à saúde, mas quando realizada em dias consecutivos pode intensificar os efeitos deletérios da fadiga muscular, podendo gerar perda de atividade neuromuscular, aumento do dano às fibras musculares e incremento do risco de lesão. Estratégias para inibir ou retardar estes efeitos da fadiga, principalmente o estresse oxidativo, são desejáveis. Nesse contexto, o extrato de chá verde (ECV), derivado da Camellia sinensis, pode ser útil, visto que tem conhecidas propriedades anti-inflamatórias e antioxidantes. Neste estudo, nosso objetivo foi verificar se a suplementação com extrato de chá verde beneficia marcadores de dano muscular, estresse oxidativo e desempenho em condição de fadiga. Dezesseis atletas amadores treinados, todos homens, foram testados durante exercício de 9 ciclismo em duas condições de exercício: sem suplementação sem fadiga; e com suplementação (placebo, n=8; extrato de chá verde 500 mg/dia durante 15 dias, n=8) com fadiga (extensão de joelhos até a exaustão induzida em dois dias consecutivos). Coletas sanguíneas para determinação de marcadores de dano muscular e estresse oxidativo, e medidas de eletromiografia, para determinação da ativação elétrica neuromuscular dos vastos laterais, além de parâmetros de treinamento e frequência cardíaca, foram quantificados e comparados entre os grupos nas diferentes condições. Os valores normalizados de eletromiografia root mean square e mediana da frequência da eletromiografia não diferiram entre a pedalada sem e com fadiga no grupo chá verde, enquanto que o grupo placebo apresentou prejuízo na contratilidade muscular decorrente da fadiga. O maior custo de frequência cardíaca confirma que esse redução na EMG foi combinada por um aumento do esforço na tarefa. O ECV também foi capaz de inibir dano muscular e evitar o estresse oxidativo, ao contrário do grupo placebo na comparação entre as pedaladas sem e com fadiga. Dessa forma, as propriedades antioxidantes do chá verde parecem evitar o dano muscular induzido pelo estresse oxidativo e preservar a contratilidade muscular de atletas submetidos à fadiga em dias consecutivos. Sendo assim, a suplementação com chá verde pode ser considerada como uma estratégia válida para o treinamento físico em tarefas extenuantes. / The regular practice of intense physical exercise benefits health, but when the sessions are accomplished in consecutive day it migh intensify some deleterious effects of muscle fatigue, which can impair efficiency, promote muscle damage and increase injury risk. The interest in strategies to avoid or delay such effects, especially the oxidative stress, are desirable. In this regard, the green tea extract, from Camellia sinesis, can be useful due to its recognized antioxidant and antiinflamatory properties. Here our purpose was to determine whether the supplementation with green tea extract can benefit markers of muscle damage, oxidative stress and performance in exercise under fatigue. Sixteen male athletes trained were evaluted in cycling exercise under two conditions: without supplementation and without fatigue, and supplemented (placebo n=8; extrato de chá verde, n=8, 500 mg/dia durante 15 dias) and with fatigue (knee extension until exhaustion in two consecutive days). Blood 11 samples served to determine markers of muscle damage and oxidative stress. Electromyography was employed to monitore the muscle activation from vastus lateralis, as well as training parameters like heart rate, were quantified and compared between the groups and exercise conditions. Muscle activation did not differ between the cycling with and without fatigue in the green tea group, whereas in the placebo group there a loss of contractility due to fatigue. A higher cost observed by the higher heart rate support that reduction in the activation in the placebo group relies on the effort associated to the performance of the cycling exercise. Green tea supplementation was able to avoid increase in the muscle damage and oxidative stress in response to the exercise compared to the placebo group. Therefore, the properties of green tea seem to delay or avoid the muscle damage resultant of oxidative stress and preserves the contractility in athletes submitted to exercise under a fatigue state. As a conclusion, the supplementation with green tea can be considered a valid strategy in the physical training during extenuant tasks.

Submitted by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2018-09-28T13:49:10Z No. of bitstreams: 1 CRISTINI ESCOBAR VIANA.pdf: 1264568 bytes, checksum: 8c87e6df98a20d981c5e1c739240eae4 (MD5) / Approved for entry into archive by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2018-09-28T13:49:26Z (GMT) No. of bitstreams: 1 CRISTINI ESCOBAR VIANA.pdf: 1264568 bytes, checksum: 8c87e6df98a20d981c5e1c739240eae4 (MD5) / Made available in DSpace on 2018-09-28T13:49:26Z (GMT). No. of bitstreams: 1 CRISTINI ESCOBAR VIANA.pdf: 1264568 bytes, checksum: 8c87e6df98a20d981c5e1c739240eae4 (MD5) Previous issue date: 2018-04-11 / A hipercolesterolemia se caracteriza pela elevação patológica da taxa de colesterol sanguínea aumentando a formação de radicais livres, o qual provoca oxidação e danos nos lipídios celulares causando crescimento da atividade inflamatória e progressão negativa da função cognitiva. O consumo de vinho tinto atua minimizando estes efeitos, inibindo a expressão de citocinas e modulando os níveis do fator neurotrófico derivado do cérebro (BDNF), da proteína cíclica de ligação ao elemento AMPc-resposta (CREB), dos microRNAs (miARN), e a ação da enzima acetilcolinesterase (AChE). Os antioxidantes, os flavonóides e os polifenóis tornaram-se as primeiras substâncias contidas no vinho tinto com efeitos benéficos comprovados em várias doenças, como a inibição da oxidação de LDL e parâmetros envolvidos na resposta inflamatória. A ingestão regular de bebidas ricas em polifenóis demonstra exercer consequências favoráveis à saúde humana, como diminuição da incidência de doenças cardiovasculares, câncer e proteção contra doenças neurodegenerativas, entre outras. Portanto, o presente estudo teve como objetivo investigar o potencial efeito neuroprotetor do vinho Ruby Carbenet em camundongos nocaute para o receptor LDLr-/- expostos à dieta hipercolesterolêmica. O vinho foi submetido a análise em vitro para quantificação de polifenóis, antocianinas, ácido cafeico, ácido gálico, epicatequina e resveratrol. Foram utilizados 40 camundongos adultos com 90 dias, de 20 a 25 gramas. Os animais foram divididos em 4 grupos (n=10) e para a indução da hipercolesterolemia foi borrifado colesterol diluído em álcool puro 96° C, sobre a dieta comercial (Puro Trato PB 22) durante 90 dias. Ao final do tratamento foi realizado teste comportamental de memória, tarefa do labirinto aquático de Morris (MWMT), logo após, o sangue foi coletado por punção cardíaca e o córtex pré-frontal e o hipocampo foram removidos para os ensaios bioquímicos. Foram determinados os níveis de lipídios plasmáticos e as citocinas pró-inflamatórias, interferon gama (IFN-γ), fator de necrose tumoral-α (TNF-α), interleucina-1β (IL-1β) e interleucina-6 (IL-6) no hipocampo e córtex pré-frontal. Também foram avaliadas a atividade da AChE e os níveis das proteínas estruturais BDNF e CREB, e os biomarcadores microRNAs. Nossos resultados mostraram que a dieta hipercolesterolemica causa um aumento nos lipídios plasmáticos, nas citocinas pró-inflamatórias e na atividade da enzima AChE nos camundongos LDLr-/-, o que foi atenuado pelo consumo de vinho tinto através da diminuição do estado inflamatório. A dieta com elevado índice de colesterol diminuiu os níveis de BDNF, CREB e microRNAs cerebrais, e o consumo moderado de vinho tinto foi capaz de reverter estas proteínas estruturais BDNF e CREB, e os biomarcadores miRNA146a e 155, promovendo a neuroproteção e a melhora da memória. O vinho Ruby Cabernet apresentou potencial antioxidante confirmado pelos seus teores de ácido gálico 135, 3 mg/L, ácido caféico 91,2 mg/L, epicatequina 242,2 mg/L e resveratrol 102,2 mg/L. O consumo moderado do vinho proporcionaram aos camundongos melhora no déficit cognitivo e plasticidade, corroborando com o resultado do teste comportamental, no qual o MWMT está relacionado com o sistema de aprendizado e memória. Com isso, verificou-se que o vinho Ruby Carbenet demonstrou efeito neuroprotetor e reduziu os níveis dos lipídios plasmáticos, modulando as citocinas, agindo na manutenção da neuroplasticidade e regulação da atividade da acetilcolinesterase em camundongos hipercolesterolêmicos. Palavras-chave: Compostos fenólicos. Déficit cognitivo. Dieta com elevado índice de colesterol. Camundongos nocaute para receptor LDLr-/-. Neuroplasticidade. / Hypercholesterolemia is characterized by the pathological elevation of the blood cholesterol level, increasing the formation of free radicals, which causes oxidation and damage to the cellular lipids causing inflammatory activity growth and negative progression of cognitive function. Red wine consumption minimizes these effects by inhibiting cytokine expression and modulating levels of brain-derived neurotrophic factor (BDNF), cyclic cAMP-binding element (CREB), microRNAs (miRNA), and the action of the enzyme acetylcholinesterase (AChE). Antioxidants, flavonoids and polyphenols have become the first substances contained in red wine with proven beneficial effects in various diseases, such as the inhibition of LDL oxidation and parameters involved in the inflammatory response. Regular ingestion of polyphenol-rich beverages has a positive effect on human health, such as reducing the incidence of cardiovascular diseases, cancer and protection against neurodegenerative diseases, among others. Therefore, the present study aimed to investigate the potential neuroprotective effect of Ruby Carbenet wine in knockout mice for the Ldlr-/- receptor exposed to the hypercholesterolemic diet. The wine was submitted to in vitro analysis for quantification of polyphenols, anthocyanins, caffeic acid, gallic acid, epicatechin and resveratrol. 40 adult mice with 90 days, from 20 to 25 grams were used. The animals were divided into 4 groups (n=10) and for the induction of hypercholesterolemia cholesterol was diluted in pure alcohol 96 ° C, on the commercial diet (Puro Trato PB 22) for 90 days. At the end of the treatment was performed memory behavioral test, Morris water maze task (MWMT), soon after, blood was collected by cardiac puncture and the prefrontal cortex and hippocampus were removed for biochemical assays. The levels of plasma lipids and proinflammatory cytokines, interferon gamma (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) in the hippocampus and prefrontal cortex. We also evaluated the activity of AChE and the levels of the structural proteins BDNF and CREB, and the biomarkers microRNAs. Our results showed that the hypercholesterolemic diet causes an increase in plasma lipids, proinflammatory cytokines and AChE enzyme activity in Ldlr-/- mice, which was attenuated by the consumption of red wine through the reduction of the inflammatory state. The high cholesterol diet decreased levels of BDNF, CREB and brain microRNAs, and moderate consumption of red wine was able to revert these structural proteins BDNF and CREB, and the miRNA146a and 155 biomarkers, promoting neuroprotection and the improvement of memory. Ruby Cabernet wine showed antioxidant potential confirmed by its content of gallic acid 135.3 mg/L, caffeic acid 91.2 mg/L, epicatechin 242.2 mg/L and resveratrol 102.2 mg/L. The moderate consumption of wine gave the mice an improvement in cognitive deficit and plasticity, corroborating with the result of the behavioral test, in which MWMT is related to the learning and memory system. Thus, it was verified that Ruby Carbenet wine demonstrated neuroprotective effect and reduced plasma lipid levels, modulating cytokines, acting to maintain neuroplasticity and regulate acetylcholinesterase activity in hypercholesterolemic mice. Keywords: Phenolic compounds. Cognitive deficit. Diet with high cholesterol. Knockout mice for Ldlr-/- receptor. Neuroplasticity.

Introdução: A ingestão de carboidrato (CHO) durante o exercício de resistência aeróbica de longa duração tem se mostrado capaz de otimizar o desempenho dos atletas. Alguns estudos recentes têm reportado também um efeito ergogênico quando a proteína (PTN) é adicionada a uma bebida carboidratada. Além disso, a coingestão tem sido relacionada com a atenuação do dano muscular, tido como fator limitante da recuperação muscular. No entanto, pouco se investigou sobre a influência dessas suplementações, comparadas a uma bebida placebo (PLA), no duatlo de distância olímpica. Objetivo: Comparar o efeito de três diferentes estratégias nutricionais de suplementação (CHO vs. CHO+PTN vs. PLA), durante um teste simulado de duatlo (TSD) olímpico, no desempenho e recuperação muscular de duatletas amadores. Métodos: Em um ensaio clínico, cruzado, randomizado e duplo-cego, treze atletas amadores do sexo masculino, com idade média de 29,7 ± 7,7 anos, participaram de três TSD consumindo: bebida carboidratada (CHO, 75 g); bebida isocalórica combinando carboidrato e proteína, na proporção 4:1 (CHO+PTN, 60,5 g CHO e 14,5 g PTN); e bebida placebo (PLA). Aos atletas informou-se apenas que o estudo visava investigar a influência de três diferentes tipos de suplementação. Após jejum de 8 h, sujeitos recebiam um café da manhã padrão, com 1,5 g/kg CHO e 45 min depois iniciavam o protocolo. As intensidades da primeira corrida (10 km) e da sessão de ciclismo (40 km) foram controladas, utilizando dados previamente coletados em testes preliminares, e a corrida final, de 5 km, foi tratada como um contrarrelógio (t5km). Coletas de sangue foram realizadas antes, imediatamente após e 24h após cada teste simulado, e analisou-se concentrações de glicose e creatina quinase (CK). Pico de torque (PT) isométrico foi mensurado no início do estudo e 24h após cada teste. Resultados: Os atletas completaram as distâncias totais do duatlo olímpico em ~1h 51min. Não houve diferença significativa no t5km entre as condições CHO (1270,3 ± 130,5 s) vs. CHO+PTN (1267,2 ± 138,9 s) vs. PLA (1275,4 ± 120 s); p = 0,87; TE ≤ 0,1. Os resultados de PT não demonstraram alterações significativas entre as condições basal (302,2 ± 52,8 N.m) vs. pós-24h CHO (300,1 ± 41,4 N.m) vs. pós-24h CHO+PTN (292,2 ± 49,4 N.m) vs. pós-24h PLA (282,1 ± 43,1 N.m); p = 0,24; TE ≤ 0,4. Embora, os resultados de CK tenham mostrado aumento significativo para todas as condições na comparação pré vs. pós-24h: CHO (300%; p < 0,01; TE = 0,93); CHO+PTN (82%; p < 0,01; TE = 0,73) e PLA (190%; p = 0,01; TE = 1,04), não foram encontradas diferenças entre as condições nos diferentes momentos - pré, imediatamente após e pós-24h (p = 0,32; TE = 0,3 – 1,04). Conclusão: Em uma prova simulada de duatlo olímpico, com refeição pré-teste contendo 1,5g/kg de carboidrato, as suplementações de CHO e de CHO+PTN não oferecem benefícios extras quando comparadas a uma bebida placebo no que diz respeito ao desempenho e recuperação muscular dos atletas. / Introduction: Carbohydrate (CHO) intake during long-term endurance exercise has been shown to optimize performance in athletes. Some recent studies have also reported an ergogenic effect when protein (PRO) is added to a carbohydrate drink. Furthermore, the coingestion has been related to the attenuation of muscle damage, seen as a limiting factor in muscle recovery. However, little has been investigated about the influence of such supplementation, compared to a placebo drink (PLA), in an Olympic-distance (OD) duathlon. Purpose: This study aimed to compare the effects of three different nutritional supplementation strategies (CHO vs. CHO+PRO vs. PLA) during a simulated OD duathlon, on performance and indices of muscle recovery of amateurs duathletes. Methods: In a crossover, randomized, double-blind clinical trial, thirteen male amateur athletes, mean age 29.7 ± 7.7 years, participated in three simulated OD duathlons consuming either a carbohydrate drink (CHO, 75 g); an isocaloric drink combining carbohydrate and protein in a 4:1 ratio (CHO+PRO, 60.5 g CHO and 14.5 g PRO); or a placebo (PLA) drink. The athletes were informed that the study aimed to investigate the influence of three different types of supplementation. After an 8 hours fast, subjects received a standard breakfast, with 1.5 g / kg CHO and 45 min later the protocol began. The intensities of the first running (10 km) and cycling session (40 km) were controlled using previously collected data, and the final running, 5 km, was treated as a time-trial (t5km). Blood samples were taken before, immediately after and 24 hours after each simulated duathlon and glucose and creatine kinase (CK) levels were analyzed. Isometric peak torque (PT) was measured at baseline and 24 hours after each test. Results: The athletes completed the OD duathlons in ~ 1h 51min. There was no significant difference in t5km between CHO (1270.3 ± 130.5 sec) vs. CHO+PRO (1267.2 ± 138.9 sec) vs. PLA (1275.4 ± 120 sec); p = 0.87; ES ≤ 0.1. The results of PT showed no significant changes between baseline conditions (302.2 ± 52.8 Nm) vs. 24h post-CHO (300.1 ± 41.4 Nm) vs. 24h post-CHO+PRO (292.2 ± 49.4 Nm) vs. 24h post-PLA (282.1 ± 43.1 Nm); p = 0.24; ES ≤ 0.4. Although CK results showed significant increase for all conditions when comparing pre vs. post 24h CHO (300%; p <0.01; ES = 0.93); CHO+PRO (82%; p <0.01; ES = 0.73) and PLA (190%, p = 0.01; ES = 1.04), no differences were found between the conditions in the different moments - pre, immediately after and after 24 hours (p = 0.32; ES = 0.3 – 1.04). Conclusion: In a simulated Olympic-distance duathlon, with pre-meal test containing 1.5g / kg of carbohydrate, the ingestion of CHO and CHO+PRO does not provide additional benefits when compared to a placebo drink regarding athletes’ performance and muscle recovery.

O objetivo da presente dissertação foi verificar os efeitos de 22 semanas de treinamento com ênfase na capacidade técnica-tática sobre o comportamento das capacidades motoras, indicadores indiretos de dano muscular [creatina quinase (CK) e lactato desidrogenase (LDH)] e o desempenho em campo de jovens futebolistas (Capítulos 2 e 3). Além disso, objetivou-se comparar o primeiro com o segundo tempo de um jogo simulado de futebol no que se refere aos padrões de deslocamento e variáveis táticas e; identificar possíveis associações entre a CK e a LDH com os padrões de deslocamento em um jogo simulado (Capítulo 4). Participaram do estudo 20 voluntários (idade 15,1 ± 0,3 anos, estatura 171,5 ± 6,1 cm, massa corporal 60,4 ± 5,5 kg, % gordura 11,5 ± 2.8, pico de velocidade de crescimento 1,42 ± 0,5 anos, VO2máx 48,68 ± 4,67 ml.kg-1.min-1). As capacidades motoras foram acompanhadas por meio dos testes motores: salto vertical e salto horizontal; T-40; Shuttle Run Test e o Running Anaerobic Sprint Test. Para determinação da atividade plasmática das enzimas CK e LDH foram realizadas coletas de sangue venoso (10ml). O desempenho em campo foi verificado por meio de análises cinemáticas bidimensionais dos padrões de deslocamento (distância total, velocidade média, número de sprints, % nas zonas de intensidade) e variáveis táticas (área de ocupação da equipe, espalhamento dos jogadores) em um jogo simulado filmado por duas câmeras captando toda a área do jogo. O desempenho nas partidas da competição alvo foi analisado por meio do aproveitamento de pontos. Todas as avaliações supracitadas foram realizadas em quatro momentos distintos: pré etapa preparatória (T0); pós etapa preparatória (T1); pós etapa competitiva I (T2); pós etapa competitiva II (T3). Para todos os resultados obtidos por meio dos testes motores foram encontradas diferenças significantes (p 0,05) quando comparado os momentos T0 (pré-treinamento) com T3 (pós-treinamento), no sentido de ganho/melhora (Capítulo 2). No desempenho das partidas foi verificado um aproveitamento de 83,33% nos pontos disputados (Capítulo 2). Em relação ao comportamento dos padrões de deslocamento e variáveis táticas ao longo da periodização, foi verificado aumento significante (p 0,05) da intensidade do jogo e do desempenho tático quando comparado o momento T0 com T3. Fato evidenciado devido o aumento das variáveis: porcentagem da distância total percorrida em sprinting, em alta intensidade (AAI), na velocidade máxima, área de ocupação e espalhamento (Capítulo 3). No que refere-se à atividade plasmática da CK e da LDH, observou-se uma redução significante (p 0,05) (Capítulo 3). Em relação ao Capítulo 4, observa-se nos resultados um aumento significante (p 0,05) da distância total, da velocidade média, do número de sprints, AAI e das variáveis táticas no segundo tempo do jogo simulado. Em adição, verificou-se diminuição significante da porcentagem da distância total percorrida em baixa intensidade no segundo tempo. Além disso, verificou-se grande associação entre a porcentagem do delta de alteração da CK e da LDH com os padrões de deslocamento no jogo simulado. Conclui-se no Capítulo 2 um aprimoramento significativo nos índices das capacidades motoras dos futebolistas estudados, além de um expressivo aproveitamento de pontos na competição alvo. No Capítulo 3 sintetiza-se que houve aumento do desempenho em alta intensidade no jogo, no rendimento tático e redução da atividade dos marcadores bioquímicos relacionados ao dano muscular. Em relação ao Capítulo 4 conclui-se que indicadores indiretos de dano muscular possuem grande associação com padrões de deslocamento em jogo realizado em condições de treino por jovens futebolistas / The purpose of the present dissertation was to verify the effects of 22 weeks of training, emphasizing the technical-tactical ability, on the behavior of motor capacities, indirect skeletal muscle damage markers [creatine kinase (CK) and lactate dehydrogenase (LDH)] and field performance of young soccer players (Chapters 2 and 3). Furthermore, it was aimed to compare the first and second halves of a simulated match regarding displacement patterns and tactical variables; and identify potential associations between the CK and LDH with the displacement patterns within the simulated match (Chapter 4). Twenty volunteers accepted to participate in this study (age 15.1 ± 0.3 years; height 171.5 ± 6.1 cm; body weight 60.4 ± 5.5 kg; body fat % 11.5 ± 2.8; peak height velocity 1.42 ± 0.5 years; VO2max 48.68 ± 4.67 ml.kg-1.min-1). The motor capacities were assessed by the motor tests: vertical jump and horizontal hump; adapted T-40; Shuttle Run Test and Running Anaerobic Sprint Test (RAST). To determine CK and LDH plasmatic activity, vein blood samples were collected (10ml). The field performance was assessed by displacement patterns kinematic two-dimensional analysis (total distance; average velocity; number of sprints; % on each intensity zone) e tactical variables (team occupation zone; players field distribution) on a simulated match, recorded by two cameras covering the whole area of the match. The performance on matches within the target competition were analyzed by earned points during the period. All the assessments mentioned were performed in four distinct moments: before preparatory stage (T0); after preparatory stage (T1); after competitive stage I (T2); and after competitive stage II (T3). When comparing moments T0 (before training program) and T3 (after training program), significant differences were found for all results regarding motor tests (p 0.05), in the direction of gain/improvement (Chapter 2). On matches performance, 83.33% of the contested points were achieved (Chapter 2). Regarding the behavior of the displacement patterns and tactical variables over the periodization, it was verified a significant improvement (p 0.05) on games intensity and tactic performance when comparing moments T0 and T3. This fact is proved by an increase of the variables: percentage of sprinting total covered distance, in high intensity, in maximal intensity, occupation and field distribution (Chapter 3). Concerning the plasmatic activity of CK and LDH, it was observed a significant reduction (p 0.05) (Chapter 3). On the Chapter 4, it is noticed on the results an significant improving (p 0.05) of total distance, average velocity, number of sprints, high intensity activities and tactical variables. Additionally, it is possible to verify a significant decreasing of low intensity total distance covered percentage on the second half. Moreover, it can be observed a large association between the percentage of CK and LDH delta variation with the displacement patterns on the simulated match. Through the Chapter 2 evidences, it is possible to conclude a significant enhancement of the motor capacities rates of the studied soccer players, besides an expressive earned points on the target competition. On Chapter 3 it is evident the performance gains during high intensity efforts along the match, on tactical performance and the reduction on biochemical markers for skeletal muscle damage. Yet, by the Chapter 4 outcomes, it is allowed to conclude that indirect skeletal muscle damage markers have large association with displacement patterns during a match performed on training conditions by young soccer players

Introdução: As aulas de ballet parecem apresentar intensidades cardiorrespiratórias (CR) mais baixas do que os ensaios e espetáculos. Além disso, o ballet é caracterizado como um exercício intermitente, que envolve uma variedade de ações excêntricas, as quais podem estar relacionadas à lesão muscular (LM) e ao estresse oxidativo (EO). Objetivos: Descrever, comparar e correlacionar as respostas CR, de EO e LM em bailarinas após uma aula e um ensaio de ballet. Variáveis CR: consumo de oxigênio (VO2); frequência cardíaca (FC) e concentração sanguínea de lactato (La). Variáveis EO: estado redox (GSSH/GSH) e concentração sanguínea de lipoperóxidos (LPO). Variável de LM: concentração sanguínea de creatina quinase (CK). Métodos: Doze bailarinas voluntárias, de nível técnico avançado foram avaliadas. Teste de consumo máximo de oxigênio (VO2máx) e avaliação da composição corporal foram realizados. Em duas sessões separadas, as bailarinas realizaram uma aula e um ensaio de ballet, os quais foram previamente filmados na escola de dança e reproduzidos nas sessões de coleta de dados. VO2 e FC foram mensurados continuamente em todas as sessões. La foi verificado antes e depois do teste de VO2máx; e em repouso, aos 15 e 30 minutos da aula e do ensaio. Coletas de sangue foram realizadas em repouso, imediatamente após e 48h após a aula e o ensaio. As variáveis CR da aula (barra, centro e aula toda) e do ensaio (ensaio todo) foram também relacionadas aos dados do primeiro e segundo limiares ventilatorios (LV1 e LV2). La pós teste VO2max, pós aula e pós ensaio foram também comparados entre si. Dados expressos em média e desvio padrão. Estatística: ANOVA Two-way; ANOVA medidas repetidas; Post hoc Bonferroni (p<0,05). Resultados: VO2máx=37,3±4,7; LV1=24,92,7 e LV2=31,9±3,8 ml.kg-1.min-1. VO2 (aula=14,5±2,1 / ensaio=19,11,7 ml.kg-1.min-1); FC (aula=145,7±17,9 / ensaio=174,5±13,8 bpm) e La (aula=4,2±1,1 / ensaio=5,5 ± 2,7 mmol.l-1) foram significativamente diferentes entre si. Resultados Os resultados do VO2 (ml.kg-1.min-1 ) comparando-se aula, ensaio, LV1 e LV2 foram: aula barra (14,4±2,0); aula toda (14,5±2,1); aula centro (16,7±2,5); ensaio todo (19,1±1,7); LV1 (24,9 ± 2,7) e LV2 (31,9 ± 3,8). Aula barra e aula toda foram iguais entre si e diferentes de aula centro e ensaio todo, os quais não foram diferentes entre si. LV1 e LV2 foram diferentes entre si e de todos os demais parâmetros. Para FC, o ensaio todo se localizou entre LV1 e LV2. La (mmol.l-1) da aula (4,2±1,1) foi significativamente menor do que o La máximo (8,1±2,3), sendo o ensaio (5,5±2,7) estatisticamente igual a ambos. CK foi significativamente mais elevada pós aula do que ensaio, sendo os valores pós e 48h iguais entre si e ambos diferentes do pré. A razão GSSG/GSH diminuiu significativamente 48h pós aula e ensaio, mas não foi diferente entre os dois tipos de exercício. Os valores de LPO foram maiores para a aula do que para o ensaio, não apresentando diferenças em relação ao efeito tempo. Conclusão: A aula apresenta intensidade mais baixa do que o ensaio em relação às variáveis CR, entretanto, a aula foi mais intensa no que se refere aos parâmetros de dano celular. Parece que as bailarinas avaliadas estão adaptadas em relação aos parâmetros de dano celular, mas necessitam de treinamento mais específico do ponto de vista CR. / Introduction: Ballet classes (BC) seem to have lower cardiorespiratory (CR) intensities than ballet reharshal (BR) and spectacles. Besides that, the ballet is characterized as an intermittent exercise that involves several eccentric avtions, which could be related to muscle damage (MD) and oxidative stress (OS). Aims: To describe, compare and correlate ballet dancers` CR, MD and OS responses after a BC and a BR. CR variables: oxygen consumption (VO2); heart rate (HR) and lactate blood concentration (La). OS variables: redox state (GSSH/GSH) and lipoperoxides blood concetration (LPO). MD variable: creatine kinase blood concentration (CK). Methods: Twelve female advanced ballet dancers volunteered this study. Maximum oxygen consumption (VO2max) test and body composition assessment were performed. In two separated sessions, the dancers performed a BC and a BR, which were previously recorded in their dance school, and further transmited during the data colecttion sessions. VO2 and HR were continually measured during all sessions. La was verified before and after the VO2max test; and in rest, in 15 and 30 minutes of the BC and BR. Blood colections were performed in rest, immediately after and 48h after the BC and BR. CR responses during BC (barre, center floor and whole BC) and during BR (whole BR) were also compared to the CR responses in the first and second ventilatory threshold (VT1 and VT2). The La was also compered after VO2max test, after the BC and after BR. Data were expressed in average and standart deviation. Statistics: ANOVA Two-way; ANOVA repeated measures and Post hoc Bonferroni (p<0.05). Results: VO2max=37.3±4.7; VT1=24.92.7 and VT2=31.93.8 ml.kg-1.min-1. VO2 (BC=14.5±2.1 / BR=19.1±1.7 ml.kg-1.min-1); HR (BC=145.7±17.9 / BR=175±13.8 bpm) e La (BC=4.2±1.1 / BR=5.5 ± 2.7 mmol.l-1) were significantly different among themselves. VO2 (ml.kg-1.min-1) results comparing BC, BR, VT1 and VT2 were: barre (14.4±2.0); whole BC (14.5±2.1); center floor (16.7±2.5); whole BR (19.11.7); VT1 (24.9 ± 2.7) and VT2 (31.9 ± 3.8). For FC results, the whole BR was located between VT1 and VT2. La (mmol.l-1) in the BC (4.2±1.1) was significantly lower than La in the VO2max test (8.1±2.3), being the BR (5.5±2.7) statistically equal to both. CK was significantly higher after the BC than the BR, being the values post and 48h post equal between themselves and both significantly different from before BC and BR. The ratio GSSG/GSH was significantly lower in 48h after BC and BR, but it was not different in relation to the type of exercise (BC or BR). LPO values were higher in the BC than the BR, however they did not show any differences related to the time. Conclusion: BC showed lower CR responses than BR, nevertheless, BC was more intense concerning the cell damage parameters. It appears that the dancers evaluated are well adapeted in relation to cell damage parameters, but they need more specific training from the CR point of view.