Made available in DSpace on 2014-07-29T15:07:44Z (GMT). No. of bitstreams: 1 Leandro_Franco.pdf: 1233771 bytes, checksum: d1a3e21430ce8cb834c7bb95cd287196 (MD5) Previous issue date: 2008-03-05 / Changes in physiological parameters of electrocardiography, echocardiography and biochemistry markers were evaluated in dogs anesthetized with different associations of atropine, xylazine and S-ketamine. Twenty three healthy female dogs randomly distributed in four groups named as GI-6, GII-6, GIII-6 and GIV-5 were treated respectively with atropine and S-ketamine (0,04mg/kg; 10 mg/kg); S-ketamine (10 mg/kg); atropine, xylazine and S-ketamine (0,04mg/kg; 1,1 mg/kg; 10 mg/kg) and xylazine and S-ketamine (1,1 mg/kg; 10 mg/kg). Ten minutes after induction, 5mg/kg of S-ketamine was administered to animals from all groups. Measurements of electrocardiogram, echocardiogram and serum activity of aspartate aminotransferase (AST), creatine kinase (CK) and creatine kinase MB isoenzyme (CK-MB) were evaluated for 36 hours. Concerned to electrocardiography, the proposed anesthetic protocols showed significant changes especially in atrioventricular conduction period (PR interval) and systole period (QT interval), mostly in GIV. In relation to echocardiography, the main alterations happened in following variables, final stroke volume, ejection fraction and cardiac output, predominantly in GIV. In serum biochemistry analysis, it was observed alterations in CK and CK-MB values in all groups, which maintained changed for a longer time in GIII and GIV. Thus, under conditions of this study, it can be conclude that atropine, xylazine and S-ketamine association (GIII) determined lower effects on heart, while marking alterations occurred in animals from GIV / Avaliaram-se as alterações eletrocardiográficas, ecocardiográficas e de marcadores bioquímicos em cadelas anestesiadas com atropina-xilazina-cetamina-S (+) em diferentes associações. Utilizaram-se 23 cadelas, clinicamente saudáveis, distribuídas aleatoriamente em quatro grupos experimentais (GI-n=6, GII-n=6, GIII-n=6 e GIV-n=5). Os animais do GI, GII, GIII e GIV foram tratados respectivamente com atropina-cetamina-S (+) (0,04 mg/kg 10 mg/kg), cetamina-S (+) (10 mg/kg), atropina-xilazina-cetamina-S (+) (0,04mg/kg 1,1 mg/kg -10 mg/kg) e xilazina-cetamina-S (+) (1,1 mg/kg -10 mg/kg). Após dez minutos da aplicação da indução, foram reaplicados 5,0 mg/kg de cetamina-S (+) nos animais de ambos os grupos. Avaliaram-se os animais por um período de 36 horas, por meio de eletrocardiograma, ecocardiograma e avaliações da atividade sérica de aspartatoaminotransferase (AST), creatinoquinase (CK) e creatinoquinase fração-MB (CK-MB). Relativamente à eletrocardiografia, os protocolos anestésicos estudados desencadearam mudanças significativas especialmente no tempo de condução atrioventricular, evidenciado pelo aumento da duração do intervalo PR, e período de sístole, evidenciado pelo aumento da duração do intervalo QT, sugerindo um quadro de sobrecarga ventricular, predominantemente evidenciados nos animais do grupo tratado com a associação xilazina-cetamina-S (+). Quanto à avaliação ecocardiográfica, as principais alterações foram evidenciadas entre as variáveis, volume sistólico final, fração de ejeção e débito cardíaco, predominantemente nos animais xilazina-cetamina-S (+). Com relação à avaliação bioquímica, notou-se que independente do tratamento adotado foram observadas alterações nos valores de CK e CK-MB, permanecendo alteradas por um período maior nos animais dos grupos tratados com atropina-xilazina-cetamina-S(+) ou xilazina-cetamina-S (+). Desse modo, diante das condições em que o estudo foi realizado permite-se concluir que dentre os grupos estudados, a associação atropina-xilazina-cetamina-S (+) desencadeou menores efeitos sobre o coração, enquanto que as alterações mais significativas ocorreram nos animais tratados com xilazina-cetamina-S(+)

A melhora na capacidade física e/ou reserva funcional de um indivíduo deve-se, entre outros, a um treinamento sistemático e bem padronizado. O exercício aeróbio é uma modalidade frequentemente empregada em um programa de treinamento, especialmente naqueles com objetivo de melhora do condicionamento físico. Contudo atualmente, o uso da fototerapia tem ganhado espaço no que tange ao aumento de desempenho de atletas. Baseado nisso o objetivo deste estudo foi verificar o efeito do treinamento unipodal, terapia LED e treinamento associado à terapia LED sobre parâmetros ergoespirométricos de performance (VO2max, AT e RCT) e na concentração sanguínea de creatina quinase. Para esse estudo participaram 24 voluntários do sexo masculino, os quais foram submetidos a uma avaliação pré-intervenção, foram submetidos a um período de oito semanas de treinamento em cicloergômetro com irradiação LED ou placebo. Após esse período foram submetidos a uma nova avaliação, pós-treino. Para a aplicação da terapia LED, foi utilizado um arranjo contendo 50 LEDs com comprimento de onda de 850 nm e aplicado por 60 segundos após cada sessão de treino. Os voluntários foram separados nos seguintes grupos experimentais: Não Treinados e LED desligado (Grupo NTLD), Não Treinados e LED ligado (Grupo NTLL), Treinados e LED desligado (Grupo TLD), Treinados e LED ligado (Grupo TLL). A perna que recebeu a intervenção foi eleita por sorteio e foi chamada de perna ativa (A) e a perna contralateral, chamada de controle (C). Durante o teste em exercício crescente, até a fadiga, foram quantificadas as variáveis ergoespirométricas, ventilação (Ve), consumo de oxigênio (VO2), equivalente ventilatório de oxigênio (EqVO2), também foi coletada amostra de sangue para dosagem da lactacidemia e foi mensurada a concentração da enzima creatina quinase (CK). Como resultados das respostas agudas frente ao ato de pedalar com uma perna notou-se que a pedalada unipodal apresentou menor valor de intensidade, VO2, Ve, lactacidemia e maior incremento de CK, quando comparada com a pedalada bipodal (convencional). Ao se concluir as oito semanas de treinamento notou-se que a perna ativa na pedalada unipodal apresentou melhoras nas variáveis mensuradas nos grupos TLL, NTLL e algumas no TLD. Também foi encontrada melhora para a perna controle, em algumas variáveis, dos grupos que receberam a terapia LED. Com esse estudo conclui-se que o treinamento unipodal é capaz de melhorar parâmetros como intensidade, ventilação, consumo de oxigênio, eficiência aeróbia, e ainda minimiza os danos musculares induzido pelo exercício no membro que efetuou o treinamento. A terapia LED é capaz de promover as mesmas melhoras em intensidades, consumo de oxigênio e eficiência aeróbia que o treinamento proporcionou, contudo em magnitudes diferentes, ainda é capaz de proporcionar efeito protetor contra lesão muscular induzida pelo exercício. A terapia LED associada ao treinamento potencializa os efeitos conseguidos com cada um isoladamente. / The improvement in physical capacity and/or functional reserve is due to, among other factors, a systematic and organized training program. Aerobic exercise is a frequently used modality in a training program, especially among those who aim to improve physical fitness. However phototherapy has currently gained space in regard to improving the performance of athletes. Based on this information, the purpose of the study was to verify the effect of one-legged cycling, LED therapy and their association on ergospirometric parameters of performance (VO2max, AT and RCT) and on blood concentration of CK. 24 male subjects volunteered for the study and were submitted to a pre-intervention evaluation and an 8-week period of one-legged cycling with LED therapy or placebo. After the intervention period subjects were submitted to a second evaluation. An arrangement containing 50 LEDs and a wavelength of 850 nm was applied during 60 seconds after each training session. Volunteers were divided into the following experimental groups: untrained and LED off (ULOf), untrained and LED on (ULOn), trained and LED off (TLOf) and trained and LED on (TLOn). The leg that received the intervention (training and/or LED therapy) was selected randomly and named active leg (A) and the contralateral limb was named control (C). During the incremental exercise test to volitional failure the ergospirometric variables ventilation (Ve), oxygen uptake (VO2) and oxygen ventilatory equivalent (EqVO2) were measured and blood samples were collected after each stage for lactate and before and 24 hours after the test for the analysis of blood CK. Acute responses from the pre-intervention evaluation showed that the intensity of one-legged cycling led to lower intensity, VO2, Ve, blood lactate and higher increase in blood CK compared to traditional cycling (two-legged). After the 8 weeks of intervention, the active leg showed improvements in the measured variables for the TLOn, ULOn and TLOf groups and, in some of the variables for the groups that received LED therapy. With these data we conclude that one-legged cycling is capable of increasing parameters such as intensity, ventilation, oxygen uptake, aerobic efficiency and minimizes the blood CK responses to exercise in the exercised limb. The association between exercise and Led therapy potentiates the effects achieved with either of the interventions.

Objectives: This study investigated chronic changes in cell free DNA (cf-DNA) throughout a collegiate soccer season. The relationship between cf-DNA, C-reactive protein (CRP), creatine kinase (CK), testosterone (T), cortisol (C), testosterone-cortisol ratio (T:C), body mass and body composition were also examined. Design: Longitudinal study design with repeated measures and group comparisons.Methods: Twenty three NCAA Division I male soccer players were divided into two groups. Starters were placed in Group 1 (G1) and non-starters were placed in Group 2 (G2). cf-DNA, CRP, CK, T, C, T:C, body mass and body composition were taken three times, corresponding to pre-season, approximately mid-season and immediately after the concluding the season.Results: In G1, cf-DNA, CRP, CK, cf-DNA %∆, CRP %∆ and, CK %∆ were all statistically higher at T2 and T3 than T1. In G2, CRP %∆ was statistically higher at T2 than T1. In G2, cf-DNA %∆, CRP %∆ and CK %∆ were higher at T2 and T3 than T1.Conclusions: This suggests that cf-DNA may be a useful marker that can reflect accumulated soccer training and competitive stressors.

The primary purposes of this dissertation were to explore relationship between cell free DNA (cf-DNA), creatine kinase (CK), C-reactive protein (CRP), vertical jump testing delayed onset muscle soreness (DOMS) in response to a high-volume resistance training protocol, and to assess the sensitivity of cf-DNA to different resistance training volume loads. The secondary purpose was to examine the relationship between cf-DNA and relative strength. Study 1 was an exploratory attempt to discover relationships between cf-DNA, CK, CRP, delayed onset muscle soreness, and performance variables. Seventeen resistance trained males were recruited, 9 were randomly assigned to receive BCAAs while 8 received a placebo. Participants performed a high-volume resistance training session consisting of the back squat and bench press. Blood was drawn to measure serum cf-DNA, CK, and CRP levels prior to the training session, with cf-DNA collected immediately post, and CK and CRP at 24hr and 48hrs post. Self-reported DOMS on a scale of 1 to 10 was collected prior to training on day 2, day 3, and day 4. SJH, CMJH, and BOSCO were collected on day 1, day 3, and day 4. Fifty-seven correlations were run to explore the relationships between variables. Only the correlation between %Δ DOMS 48hr and %Δ CRP 48hr in the non-supplement group was significant (p = 0.02). The second study, designed to assess the sensitivity of cf-DNA to different resistance training volume loads, consisted of a high-volume resistance training protocol. Blood was drawn immediately before the resistance training session (T1), immediately after the third lifting set (T2), and immediately after the sixth lifting set (T3). cf-DNA increased significantly from T1 to T2 (p < 0.01) and T1 to T3 (p < 0.01). The linear regression model used to examine the capabilities of relative strength to predict %Δ cf-DNA from T1 to T3 was significant (p = 0.04). The results of this study demonstrate the short response time of cf-DNA in relation to variations in resistance training volume-load, suggesting it may be a valuable marker in monitoring the immune response to volume-load. Results also demonstrated the positive relationship between relative strength and %Δ cf-DNA.

Dita Kašparová Chronická hypoxie a exprese genů 4 Abstract Adaptation to chronic hypoxia (CH) is characterized by a variety of functional changes in order to maintain metabolic and energy homeostasis. It has been known for many years that both humans and animals indigenous or adapted to high-altitude hypoxia are more tolerant to an acute ischemic injury of the heart. Cardioprotective mechanisms activated by adaptive responses to chronic hypoxia can be the result of altered transcriptional regulations in left ventricles. Here we report results from the gene expression profiling of adaptive responses in three models of chronically hypoxic heart. Adult male Wistar rats were exposed for 21 days to either continuous normobaric hypoxia (CCH; 10% O2) or CCH interrupted daily by 1-hour reoxygenation (RCH) or CCH interrupted daily by 16-hour (CIH). Cardiprotective effect of CCH adaptation is abolished by brief daily reoxygenation, RCH adaptation. In the present study, we aimed to determine myocardial mRNA expression of 19 candidate genes divided into three important groups: i) Hypoxia inducible factor (HIF1α) and its prolyl and asparaginyl hyroxylases (PHDs and FIH respectively, ii) Creatine kinase (CK) isoenzymes which play important role in energy homeostases of heart and iii) the group of main enzymatic...

Massage is often recommended to athletes to facilitate recovery and attenuate DOMS. The purpose of this study was to investigate the effects of massage on perceived muscle soreness and pain, inflammatory and immune markers, ROM, and mood state. Fourteen, recreationally active, women participated in a randomized crossover design study, consisting of 1) 60 min. full body massage following unaccustomed exercise and 2) 60 min. of rest. following unaccustomed exercise. Perceived muscle soreness and pain, active range of motion (ROM), mood state, along with blood concentrations of interleukin-6 (IL-6), C-reactive protein (CRP), creatine kinase (CK), and neutrophil count (NC), was assessed at baseline, 4hrs, and 24hrs following both treatment and control conditions. The aims of this study were 1) to decrease the effects of delayed onset muscle soreness (DOMS), and increase time to recover, and 2) to investigate the effect of massage vs. passive rest on inflammatory and immune markers within the blood. We hypothesized 1) an increase in ROM, a decrease in perceived physical soreness and perceived physical pain, as a result of the massage, compared to control, and 2) a decrease in blood plasma inflammatory markers, CRP, NC, CK, and IL-6, as a result of the massage, compared to control. We found massage following exercise to 1) significantly decreased perceived pain (p=0.001), 2) significantly increased immune iv markers (WBC (p=0.012) and NC (p=0.012)), and 3) significantly decreased ROM (p=0.02), compared to control. Massage had no impact on inflammatory markers (IL-6, CRP, and CK), or mood.

Det finns många biotekniska analys- och detekteringsmetoder. Metoderna används för identifiering och kvantifiering av biomarkörer. Denna studie har analyserat detekteringsmetoder i de fall där två hjärtspecifika biomarkörer används, troponin och kreatinkinas. Studien avsåg att först identifiera tillämpningsfrekvensen av detekteringsmetoder i Sverige samt internationellt. Vidare identifieras sambandet mellan avgörande parametrar i val av detekteringsmetod. Metoden gick ut på att först bestämma den mest frekventa detekteringsmetoden i Sverige med hjälp av en enkät som skickades till olika laboratorier, sedan studerades tidigare studier publicerade på olika internationella databaser. Studierna som tillämpades var på hjärtspecifika troponin och kreatinkinas för att identifiera val av detekteringsmetod, detekteringskaraktäristika och användarvänlighetsparametrar. Studiens resultat visade att nationellt finns det tre detekteringsmetoder som är de mest använda för identifiering av kreatinkinas: masspektrometri, elektrokemisk luminescence och spektrometri. Internationellt är den dominerande metoden däremot elektrokemisk luminescence. För troponin är den dominerande metoden nationellt: elektrokemisk luminescence och flödescytometri, medan internationellt: elektrokemisk luminescence. Elektrokemisk luminescence är i många fall en stark vinnare i tillämpningen. Ytterligare iakttogs korrelationskoefficienter mellan parametern för att identifiera det starkaste respektive svagaste sambandet. Avgörande parametrar i val av elektrokemisk luminescence, visar på flera samband. Elektrokemisk luminescence och kreatinkinas tilldelas en korrelationskoefficient nära ett för parametrar som volym och känslighet och en korrelationskoefficient nära minus ett för linjärt mätområde och volym, samt kostnad och minimummängd. Medan för troponin och elektrokemisk luminescence erhålls en korrelationskoefficient nära ett för parametrar som känslighet och kostnad och en koefficient nära minus ett för kostnad och tid. / There are many biotechnological analysis- and detection methods. The methods are used for identification and quantification of biomarkers. This study has analyzed detection methods incases where two heart-specific biomarkers are used, troponin and creatine kinase. The study was intended to first identify the application frequency of detection methods in Sweden and internationally. Then identify the relationship between crucial parameters in the choice of detection method. The method consisted of first determining the most frequent detection method in Sweden with the help of a questionnaire that was sent to different laboratories, then previous studies published on various international databases were observed. The studies applied were on topics regarding cardiac-specific troponin and creatine kinase to identify choice of detection method, detection characteristics, and ease of use parameters. The results of the study showed that nationally, the detection methods most used for creatine kinase are mass spectrometry, electrochemical luminescence, and spectrometry. Internationally, however, the dominant method is electrochemical luminescence. For troponin, on a national level the dominant methods are electrochemical luminescence and flow cytometry, while internationally: electrochemical luminescence. Electrochemical luminescence is in many cases a strong winner in application. In addition, correlation coefficients are observed between the decisive parameters for a detection method, to identify the strongest and weakest relationships. Electrochemical luminescence and creatine kinase are assigned a correlation coefficient close to one for parameters such as volume and sensitivity and a correlation coefficient when minus one for measurement range and volume, as well as cost and minimum amount. While for troponin and electrochemical luminescence, a correlation coefficient close to one is obtained for parameters such as sensitivity and cost and a coefficient close to minus one for cost and time.

Les isoenzymes de nucléoside diphosphate kinase (NDPK) sont connues depuis maintenant presque 60 ans et n'ont été considérées que pour leur activité catalytique de transfert de groupement phosphoryle. La découverte du gène nme, un gène antimétastatique codant une NDPK, a renouvelé l'intérêt scientifique pour cette famille d'enzymes. Il est désormais connu que la multiplication des gènes durant l'évolution a été accompagnée de diversifications structurales et fonctionnelles. J'ai étudié la fixation des NDPK-A, -B et –D (retrouvées associées aux membranes biologiques, bien que le rôle de cette association soit encore méconnu) à des membranes modèles, et j'ai trouvé des différences dans les mécanismes de fixation. J'ai montré la capacité de la NDPK-D, isoforme mitochondriale, à interagir avec des membranes anioniques ou zwitterioniques, à augmenter leur fluidité et à former des domaines protéolipidiques en présence de CL, lipide anionique spécifique de la membrane mitochondriale interne. J'ai observé cette capacité à former des domaines protéolipidiques avec d'autres protéines interagissant avec la CL, comme la créatine kinase mais pas le cytochrome C. La NDPK-A ne se fixe pas aux phospholipides du feuillet interne de la membrane plastique, ce qui suggère un autre partenaire in vivo. La NDPK-B n'interagit qu'avec des membranes anioniques via un processus en deux étapes, provoque une diminution de fluidité et est capable de former des domaines protéolipidiques. La ségrégation des lipides anioniques induite par la fixation de protéines pourrait contribuer à la formation de plateformes au sein de la membrane susceptibles de servir de point d'ancrage à de nombreuses molécules, modulant ainsi les fonctions cellulaires / Nucleoside diphosphate kinase isoenzymes (NDPK) have been known for nearly 60 years and, until recently, have been considered as housekeeping enzymes. The discovery of a nme gene, an antimetastatic gene that codes for a NDPK, revived the interest for this family. It is now known that the multiplication of nme genes throughout evolution has been accompanied with structural and functional diversification. I studied the binding of NDPK-A, -B and –D (which ae retrieved associated to cellular membranes where they are thought to play several roles) to model membranes and found differences in their behavior towards different compositions of phospholipids. I showed the ability of the NDPKD mitochondrial isoform to interact with both anionic and zwitterionic membranes, to modify their fluidity and to form proteolipidic domains in presence of CL, a mitochondrial inner membrane specific anionic lipid. I observed this ability to form proteo-cardiolipin domains with other CL interacting protein like creatine kinase but not with cytochrome c. NDPK-A was not able to bind to inner leaflet plasma membrane mimicking systems suggesting another partner in vivo. Concerning NDPK-B, it interacted only with anionic membranes via a two step-process, induced a decrease of the membrane fluidity and was able to form proteolipidic domains. Such anionic lipid segregation triggered by protein binding may contribute to platforms formation within membranes. Those platforms are then susceptible to provide a functional docking platform for numerous molecules and thus to modulate cellular functions

No description available.

Au cours de cette thèse, nous avons étudié deux protéines par RMN : la créatine kinase musculaire (CK-MM) et le domaine UIM-SH3 de la protéine STAM2, seuls ou en interaction avec leurs partenaires. La CK-MM est une enzyme active sous forme dimérique. Elle appartient à la famille des guanidino-kinases et intervient dans le processus énergétique de la cellule. Le but de l’étude était d’élucider le mode de fonctionnement de la CK-MM. Pour cela, nous avons enregistré des expériences de relaxation R1, R2 et des expériences de perturbation de déplacement chimique sur la CK-MM libre et complexée avec MgADP et sous forme TSAC. Ces expériences montrent que la boucle 320s, spécifique à la reconnaissance des substrats, possède une dynamique rapide en absence de substrats et une dynamique ralentie en présence de substrats. La fixation des substrats dans les sites actifs de la CK-MM induit des modifications conformationelles importantes. La protéine STAM2 est composée de deux UBDs : VHS, et UIM et d’un domaine SH3 connu pour interagir avec des déubiquitinases UBPY et AMSH. Cette protéine est impliquée dans la voie de dégradation lysosomale. L’objectif de cette étude est la caractérisation du complexe SH3/ubiquitine. Pour cela, nous avons enregistré des expériences de perturbation de déplacement chimique et de relaxation R1, R2 et nOes sur le complexe UIM-SH3/ubiquitine. Ces expériences mettent en évidence que les domaines UIM et SH3 sont capables d’interagir chacun avec une ubiquitine, avec une affinité de l’ordre de la centaine de micromolaire. L’interface entre les UBDs et l’ubiquitine implique majoritairement des résidus hydrophobes et conservés / In this thesis, we study two proteins by NMR: the muscular creatine kinase (CK-MM) and the SH3 domain of STAM2 protein, in the free and complexed forms. CK-MM is an active homodimeric enzyme which belongs to the guanidino-phosphagen-kinase family. This enzyme is involved in energetic process in the cell. The aim of this study is to elucidate the functional mode of the CK-MM. For this purpose, we measured R1 and R2 relaxation rates and chemical shit perturbation experiments on the substrate-free CK-MM, the CK-MM/MgADP complex, and the inhibitory ternary complex CK-MM/MgADP-creatine-nitrate. The experiments show that the loop 320s, specific recognition of the substrates, possesses a fast dynamic in absence of substrates (in the order of nano-picosecond) and a slower dynamic in presence of creatine-MgADP-nitrate ion. The binding of the substrate in the two active sites induces of significant conformational modification of the CK-MM. STAM2 protein consists in two ubiquitin binding domains (VHS and UIM) and a SH3 domain which interacts with deubiquinating enzymes AMSH and UBPY. This protein is involved in the lysosomal degradation pathway. The aim of this study is the characterization of the interaction between SH3 domain of STAM2 and ubiquitin. For this, we recorded the R1, R2, nOes relaxation experiments and chemical shift perturbation experiments on the UIM-SH3/ubiquitin complex. These experiments show that SH3 and UIM domains interact each with a single ubiquitin, with affinity of the order of hundred micromolars. The interface between these UBDs and ubiquitin, involves mainly hydrophobic and conserved amino-acids