Protein nitration is considered as a marker of reactive nitrogen species formation. Heme oxygenases (HOs) are important for the defence against oxidative stress. We evaluated the involvement of the neuronal (nNOS), the endothelial (eNOS), and the inducible (iNOS) in nitrotyrosine formation and localitzation, and both the expression and funcional significance (HO inhibition and contractility studies) of HOs in sepsis-induced muscle contractile dysfunction. Sepsis was elicited by injecting rats and transgenic mice deficient in either nNOS, eNOS, or iNOS isoforms with E.Coli lipolysaccharide (LPS). Nitrotyrosine formation and HO expressions were assessed by immunoblotting. Oxidative stress was assessed measuring protein oxidation, lipid peroxidation, and muscle glutathione. We conclude that protein tyrosine nitration occurs in normal muscles, and sepsis-mediated increase in nitrotyrosine formation is limited to the mitochondria and membrane muscle fractions. The iNOS isoform is mostly involved in nitrotyrosine formation. HOs protect normal and septic muscles from the deleterious effects of oxidants. / En un model de sepsi de disfunció diafragmàtica, s´ha avaluat el paper de les sintetases de l'òxid nítric (NOS) en la formació i localitzacio de 3-nitrotirosina, i l´expressió i significat biològic de les hemo oxigenases (HOs) (inhibidor de les HOs i estudis de contractilitat) davant l' estrès oxidatiu. La sepsi s'induí mitjançant injecció de 20 mg/kg del lipolisacàrid (LPS) d´Escherichia Coli a rates, i a ratolins deficients en les NOS induïble (iNOS), neuronal (nNOS) i endotelial (eNOS). Les proteïnes nitrificades i les HOs es van detectar amb anticossos específics. L' estrès oxidatiu s' avaluà mitjançant l' oxidació proteica, la peroxidació lipídica i el glutation muscular. Concloem que hi han proteïnes nitrificades en el múscul normal i aquestes s'incrementen durant la sepsi en les fraccions mitocondrial i membranar. L'isoforma iNOS és majorment responsable de la formació de nitrotirosina. Les HOs protegirien el múscul normal i sèptic dels efectes deleteris dels oxidants.

La Enfermedad Pulmonar Obstructiva Crónica (EPOC), presenta signos y síntomas sistémicos que se han venido explorando desde hace algún tiempo. La presente Tesis Doctoral estudia la disfunción que presentan los músculos respiratorios y periféricos, específicamente el cuádriceps de los pacientes con EPOC. Esta disfunción muscular afecta las actividades de la vida diaria, la tolerancia al ejercicio, limita la calidad de vida y disminuye la expectativa de vida de estos pacientes. Una de las posibles causas de disfunción muscular en los pacientes con EPOC puede ser la pérdida de masa muscular; por eso nos dedicamos a evaluar, los datos antropométricos de los pacientes con EPOC que asistieron durante 2 años a nuestro Laboratorio de Función Respiratoria del Hospital del Mar de Barcelona. La prevalencia de bajo peso en pacientes con EPOC en un hospital con predominante población mediterránea con, índice de masa corporal (IMC) por debajo de 20 Kg/m2, fue de tan sólo 6,6%, cifra que se reducía al 3,1% si el dintel escogido era de 18 Kg/m2. Estas cifras van claramente en oposición a la prevalencia referida en el mundo anglosajón y del norte-centro europeo, que muestra una prevalencia de bajo peso en pacientes con EPOC que oscila entre el 25 y el 35%. Teniendo en cuentas estos datos de baja prevalencia de bajo peso en nuestros pacientes con EPOC, no se explica la alta prevalencia de disfunción muscular en ellos. Por ello evaluamos variables de función muscular en pacientes con EPOC grave, específicamente del músculo cuádriceps y encontramos que los pacientes con EPOC tenían una disminución tanto de la fuerza muscular (43%), como de la resistencia muscular (77%), al compararlos con controles del mismo grupo etáreo.Tratando de profundizar en la causa de la disfunción muscular periférica en los pacientes con EPOC, hicimos a continuación otro estudio donde evaluamos el daño sarcoplásmico y sarcomérico mediante inmunohistoquímica y microscopía electrónica. Este trabajo demostró por primera vez que los pacientes con EPOC presentan mayor daño a nivel del músculo cuádriceps que los controles sanos.Debido a que en la EPOC los músculos periféricos, específicamente los de las extremidades inferiores, pudieran estar afectados por el sedentarismo, decidimos estudiar otras causas de disfunción muscular en un grupo de músculos en que este factor se hallara ausente, como son los músculos respiratorios. Estos mantienen su nivel de actividad normal o incluso aumentada. Para ello tomamos un músculo como el intercostal externo, que es fácilmente accesible a la biopsia, siguiendo un modelo mínimamente invasivo descrito por nuestro grupo. Como en trabajos precedentes ya habíamos valorado el daño sarcomérico, el estrés oxidativo o la actividad enzimática, en esta ocasión el trabajo se centró en la evaluación de la actividad inflamatoria. En él se ha demostrado que las citocinas proinflamatorias TNF-α e IL-6 se encuentran aumentadas en los músculos intercostales externos de los pacientes con EPOC al compararlo con los controles sanos. / Chronic Obstructive Pulmonary Disease (COPD), shows systemic sign and symptoms that have been studied for some time. The present Doctoral Thesis studies the dysfunction shown by the respiratory and peripheral muscles, specifically the quadriceps muscle of patients with COPD. This muscle dysfunction affects the activities of daily living, tolerance to exercise, limits quality of life and diminishes life expectancy of these patients. One of the possible causes of muscle dysfunction in the patients with COPD could be the loss of muscle mass; for that reason we evaluated, the anthropometrics data of the patients with COPD that attended during 2 years our Respiratory Function Laboratory, Hospital del Mar of Barcelona. The prevalence of low weight in patients with COPD in a hospital with a predominantly Mediterranean population with an Body Mass Index (BMI) below 20 Kg/m2, was only 6.6%, a figure that was reduced to 3.1% if the chosen threshold was 18 Kg/m2. These numbers starkly contrast to the prevalence in Northern Europe, with a low weight in patients with COPD raging from 25 to 35%. Considering these data of low prevalence of low weight in our patients with COPD, the high prevalence of muscle dysfunction is not explained. We evaluated variables of muscle function in patients with severe COPD, specifically the quadriceps muscle and we found that the patients with COPD had a decreased muscle strength (43%), and muscle endurance (77%), when comparing with healthy age matched. To study the cause of the peripheral muscle dysfunction in patients with COPD, we began another study where we assessed sarcoplasmic and sarcomeric damage by immunohystochemical methods and electronic microscopy. This work demonstrated for the first time that patients with COPD show greater muscle injury at the quadriceps muscle level that healthy age matched controls.Because in COPD, peripheral muscles, specifically those of the lower limbs, could be affected by sedentarism, we decided to study other causes of muscle dysfunction in a muscle group in which this factor was absent, as they are the respiratory muscles. These maintain their level of normal activity or activity is even increased. For this we chose a respiratory muscle like the external intercostal muscle, that is easily accessible by biopsy, following a minumum invasive model described by our group. As in preceding works of our group we evaluated the sarcomeric damage, oxidative stress or the enzymatic activity, the present work was focused in the inflammatory activity evaluation. We demonstrated that proinflammatory cytokines such as TNF-α and IL-6 are increased in the external intercostal muscles of patients with COPD when comparing with healthy age matched controls.

Made available in DSpace on 2016-06-02T20:19:15Z (GMT). No. of bitstreams: 1 3433.pdf: 978848 bytes, checksum: db91e12388088bbce1f290e9a7aeb7b9 (MD5) Previous issue date: 2011-02-04 / Financiadora de Estudos e Projetos / The dissertation consisted of two works. The aim of the first study was to evaluate respiratory muscle endurance (RME) in post myocardial infarction (MI) patients without respiratory muscle weakness [maximal inspiratory pressure (MIP)&#8805;60% of predicted] and its correlation with exercise tolerance. Participated in this study ten recent post-MI (<1.5 month post-MI [RIG]), nine late post-MI (>6 months post-MI [LIG]) and twelve healthy (control group [CG]) middle-aged men. They underwent spirometric evaluation, cardiopulmonary exercise test and two RME protocols (incremental [IP] and constant pressure [CP]). There were differences (p&#61603;0.05) in endurance pressure (PTHMAX) between the RIG and CG. There was a positive correlation for RIG between PTHMAX and maximal voluntary ventilation (MVV) (r=0.67) and peak oxygen consumption (VO2peak) (r=0.59). We conclude that RME seems to be reduced and related to peak exercise tolerance in recent MI patients without inspiratory muscle weakness. Similarly, in the second study, with nine subjects in RIG, ten in LIG and ten in CG, the aim was to characterize the metabolic and ventilatory behavior during testing of RME with IP and CP. In 70 and 80% of MIP (IL) and in the middle and final of CP test, the RIG had lower MET and higher VE/VCO2 compared CG. At 90% of MIP (IL), the RIG had lower VO2 and MET compared to CG. Moreover, RIG achieved lower pressure and product pressure-time using a similar percentage of VO2peak and oxygen consumption at anaerobic threshold (VO2AT) to sustain a 20% lower pressure than the CG. We conclude that the reduced capacity of RME in recent MI patients without inspiratory muscle weakness seems to occur due to lower ventilatory efficiency in the second part of the protocols, even in the presence of lower pressure overload. / A dissertação constou de 2 estudos descritos a seguir. O estudo I teve por objetivo avaliar a resistência dos músculos respiratórios (RMR) de indivíduos pós-infarto do miocárdio (IM) sem fraqueza muscular inspiratória [pressão inspiratória máxima (PIMAX)&#8805;60% do predito] e sua correlação com a tolerância ao exercício. Participaram deste estudo dez homens pós-IM recente (<1 mês e meio pós-IM [GIR]), nove pós-IM tardio (>6 meses pós-IM [GIC]) e doze homens saudáveis (grupo controle [GC]) de meia-idade. Estes foram submetidos a espirometria, teste cardiopulmonar e a dois protocolos de RMR [pressão incremental (RPI) e constante (RPC)]. Houve diferença (p&#61603;0,05) na pressão de resistência (PTHMAX) entre o GIR e o GC. Houve correlação positiva no GIR entre PTHMAX e ventilação voluntária máxima (VVM) (r=0,67) e o consumo de oxigênio no pico de exercício (VO2pico) (r=0,59). Concluímos que a RMR parece estar reduzida e relacionada a tolerância no pico do exercício em indivíduos pós-IM recente sem fraqueza muscular respiratória. Também foi realizado o estudo II, com nove indivíduos no GIR, dez no GIC e dez no GC, que teve por objetivo caracterizar o comportamento ventilatório e metabólico durante os testes de RPI e RPC. Em 70 e 80% da PIMAx (RPI) e no meio e final do teste de RPC, o GIR apresentou menor MET e maior VE/VCO2 em relação ao GC. Em 90% da PIMAx (RPI), o GIR apresentou menor VO2 e MET em relação ao GC. Além disto, GIR apresentou menor pressão e produto pressãotempo de RMR, utilizando semelhante percentual do VO2pico e do consumo de oxigênio no limiar de anaerobiose (VO2LAV) para pressões 20% menores em relação ao GC. Concluímos que a redução da capacidade de RMR em indivíduos na fase recente pós-IM e sem fraqueza muscular inspiratória parece ocorrer em virtude da menor eficiência ventilatória na segunda metade dos protocolos, mesmo na presença de menor sobrecarga pressórica.

Made available in DSpace on 2014-12-17T15:16:06Z (GMT). No. of bitstreams: 1 ThaiseLA.pdf: 779453 bytes, checksum: fe87c118f2c753bc8ffd4a32cb6a1146 (MD5) Previous issue date: 2009-07-22 / Background: The myotonic dystrophy (MD) is a multisystem neuromuscular disease that can affect the respiratory muscles and heart function, and cause impairment in quality of life. Objectives: Investigate the changes in respiratory muscle strength, health-related quality of life (HRQoL) and autonomic modulation heart rate (HR) in patients with MD. Methods: Twenty-three patients performed assessment of pulmonary function, sniff nasal inspiratory pressure (SNIP), the maximal inspiratory (MIP) and expiratory (MEP) pressure, and of HRQoL (SF-36 questionnaire). Of these patients, 17 underwent assessment of heart rate variability (HRV) at rest, in the supine and seated positions. Results: The values of respiratory muscle strength were 64, 70 and 80% of predicted for MEP, MIP, and SNIP, respectively. Significant differences were found in the SF-36 domains of physical functioning (58.7 ? 31,4 vs. 84.5 ? 23, p<0.01) and physical problems (43.4 ? 35.2 vs. 81.2 ? 34, p<0.001) when patients were compared with the reference values. Single linear regression analysis demonstrated that MIP explains 29% of the variance in physical functioning, 18% of physical problems and 20% of vitality. The HRV showed that from supine position to seated, HF decreased (0.43 x 0.30), and LF (0.57 x 0.70) and the LF/HF ratio (1.28 x 2.22) increased (p< 0.05). Compared to healthy persons, LF was lower in both male patients (2.68 x 2.99) and women (2.31 x 2.79) (p< 0.05). LF / HF ratio and LF were higher in men (5.52 x 1.5 and 0.8 x 0.6, p <0.05) and AF in women (0.43 x 0.21) (p< 0.05). There was positive correlation between the time of diagnosis and LF / HF ratio (r = 0.7, p <0.01). Conclusions: The expiratory muscle strength was reduced. The HRQoL was more impaired on the physical aspects and partly influenced by changes in inspiratory muscle strength. The HRV showed that may be sympathetic dysfunction in autonomic modulation of HR, although with normal adjustment of autonomic modulation during the change of posture. The parasympathetic modulation is higher in female patients and sympathetic tends to increase in patients with longer diagnosis / Introdu??o: A distrofia miot?nica (DM) ? uma doen?a neuromuscular multissist?mica que pode afetar a musculatura respirat?ria e a fun??o card?aca, e ocasionar preju?zos na qualidade de vida. Objetivos: Investigar as altera??es na for?a muscular respirat?ria, qualidade de vida relacionada ? sa?de (QVRS), e modula??o auton?mica da freq??ncia card?aca (FC) em pacientes com DM. M?todos: Foram avaliados 23 pacientes quanto ? fun??o pulmonar, press?o inspirat?ria nasal sniff (SNIP), press?es respirat?rias m?ximas (PIm?x e PEm?x), e QVRS (question?rio SF-36). Destes, 17 realizaram avalia??o da variabilidade da frequ?ncia card?aca (VFC) em repouso, nas posturas supina e sentada. Resultados: Os valores da for?a muscular respirat?ria foram de 64, 70 e 80%predito para PEm?x, PIm?x, e SNIP, respectivamente. Foi encontrada diminui??o significativa nos dom?nios do SF-36 capacidade funcional (58.7 ? 31,4 vs. 84.5 ? 23, p<0.01) e disfun??o f?sica (43.4 ? 35.2 vs. 81.2 ? 34, p<0.001) comparado a valores de refer?ncia. A an?lise de regress?o linear mostrou que a PIm?x explica 29% da vari?ncia na capacidade funcional, 18% na disfun??o f?sica e 20% na vitalidade. A VFC mostrou que, da postura supina para a sentada, o espectro AF diminuiu (0.43 x 0.30) e o espectro BF (0.57 x 0.70) e a raz?o BF/AF (1.28 x 2.22) aumentaram, com p<0.05. Comparado a valores de refer?ncia, BF foi inferior (p<0.05) tanto nos pacientes homens (2.68 x 2.99), como nas mulheres (2.31 x 2.79). A raz?o BF/AF e o espectro BF foram maiores nos homens (5.52 x 1.5 e 0.8 x 0.6), e o espectro AF, nas mulheres (0.43 x 0.21), com p<0.05. Houve correla??o significativa positiva entre tempo de diagn?stico e raz?o BF/AF (r= 0.7, p< 0.01). Conclus?es: Indiv?duos com DM t?m for?a muscular expirat?ria diminu?da. A QVRS mostrou-se mais prejudicada em rela??o a aspectos f?sicos e parcialmente influenciada por varia??es na for?a muscular inspirat?ria. Pode haver disfun??o simp?tica na modula??o auton?mica da FC, com ajuste normal da postura supina para a sentada. A modula??o parassimp?tica ? superior em pacientes mulheres e a modula??o simp?tica tende a aumentar nos pacientes com maior tempo de diagn?stico

Made available in DSpace on 2014-12-17T15:16:09Z (GMT). No. of bitstreams: 1 FernandaG_DISSERT.pdf: 1166474 bytes, checksum: 42a23625c719bb1096b1c812ac17e8f4 (MD5) Previous issue date: 2010-05-18 / The clinical importance of evaluating the respiratory muscles with a variety of tests has been proposed by several studies, once that the combination of several tests would allow a better diagnosis and therefore, a better clinical follow of disorders of the respiratory muscles. This study aimed to evaluate the feasibility of adapting a national electronic manovacuometer to measure the nasal inspiratory pressure (study 1) and analyze the level of load intensity of maximum voluntary ventilation, as well as the variables that may influence this maneuver in healthy subjects (study 2). We studied 20 healthy subjects by a random evaluation of two measures of SNIP in different equipments: a national and an imported. In study 2 it was analyzed the intensity of the load of MVV test, change in pressure developed during the maneuver, the possible differences between genders, and the correlations between the flow developed in the test and the result of MVV. In study 1 it was found the average for both measures of nasal inspiratory pressures: 125 ? 42.4 cmH2O for the imported equipment and 131.7 ? 28.7 cmH2O for the national one. Pearson analysis showed a significant correlation between the average, with a coefficient r = 0.63. The average values showed no significant differences evaluated by paired t test (p> 0.05). In the Bland-Altman analysis it was found a BIAS = 7 cmH2O, SD 32.9 and a confidence interval of - 57.5 cmH2O up to 71.5 cmH2O. In the second study it was found significant differences between the genders in the air volume moved, being higher in males 150.9 ? 13.1 l / min vs 118.5 ? 15.7 L / min for (p = 0.0002, 95% CI 44.85 to 20:05). Regarding the inspiratory and expiratory loading, they were significantly higher in men than in women, peak inspiratory pressure (34.7 ? 5.3 cmH2O vs 19.5 ? 4.2 cmH2O, 95% CI - 18.0 to -12.3, p <0.0001), peak expiratory (33.8 vs. 23.1 ? 5.9 cmH2O ? 5.4 cmH2O, 95% CI -17.1 to - 4.6, p <0.0001), and the delta pressure (59.7 ? 10 cmH2O vs 36.8 ? 8.3 cmH2O, 95% CI 14.5 to 31.2, p <0.0002). The Pearson correlation showed that the flow generated by the maneuver is strongly correlated with the delta-expiratory pressure / inspiratory (r2= 0.83,R = 0.91, 95%IC 0.72 a 0.97 e p< 0.0001).Through these results we suggest that the national electronic manovacuometer is feasible and safe to perform the sniff test in healthy subjects. For the MVV, there are differences between the genders in the intensity of pressure developed during the maneuver. We found a load intensity considered low during the MVV, and found a strong correlation between the flow generated in the test and the delta pressure expiratory / inspiratory / A import?ncia clinica de avaliar os m?sculos respirat?rios com uma variedade de testes vem sendo proposta por diversos trabalhos, pois a combina??o de v?rios testes possibilitaria um melhor diagn?stico e consequentemente um melhor seguimento cl?nico das disfun??es dos m?sculos respirat?rios. Este trabalho teve como objetivo avaliar a viabilidade da adapta??o do manovacu?metro nacional eletr?nico para a realiza??o da medida de press?o inspirat?ria nasal (estudo 1) e analisar o n?vel de intensidade de carga da ventila??o volunt?ria m?xima, assim como as vari?veis que possam influenciar a manobra em sujeitos saud?veis (estudo 2). Foram estudados 20 sujeitos saud?veis atrav?s da avalia??o aleat?ria de duas medidas de SNIP em equipamentos diferentes: um nacional e outro importado. No estudo 2 foi analisado a intensidade da carga do teste da ventila??o volunt?ria m?xima(VVM), a varia??o de press?o desenvolvida durante manobra, as poss?veis diferen?as entre g?neros, e as correla??es entre o fluxo desenvolvido no teste e o resultado da VVM. No estudo 1 as m?dias encontradas durante as duas medidas das press?es nasais foram de 125 ? 42,4 cmH2O para o aparelho importado e de 131,7 ? 28,7 cmH2O para o nacional. A an?lise de Pearson demonstrou uma correla??o significativa entre as m?dias com um coeficiente r=0.63. Os valores m?dios n?o apresentaram diferen?as significativas avaliadas pelo teste t pareado (p>0,05).Na an?lise de Bland-Altman foi encontrado um BIAS igual a 7 cmH2O, desvio padr?o 32,9 e um intervalo de confian?a de - 57,5cmH2O at? 71,5 cmH2O. No estudo 2 foi encontrada diferen?a significativa entre os g?neros para o volume de ar deslocado maior em homens 150.9 ? 13.1 l/mim vs 118.5 ? 15.7 L/mim para (p= 0.0002, IC95% 20.05 ? 44.85). Em rela??o carga inspirat?ria e expirat?ria foram significativamente maiores em homens que nas mulheres: pico inspirat?rio (34.7 ? 5.3 cmH2O vs 19.5 ? 4.2 cmH2O, IC95% - 18.0 a -12.3, p< 0.0001), pico expirat?rio (33.8 ? 5.9 cmH2O vs 23.1 ? 5.4 cmH2O, IC95% -17.1 a - 4.6, p< 0.0001), assim como o delta de press?o (59.7 ? 10 cmH2O vs 36.8 ? 8.3 cmH2O, IC95% 14.5 a 31.2 , p< 0.0002). A correla??o de Pearson mostrou que o fluxo gerado pela manobra est? fortemente correlacionado com o delta de press?o expirat?ria/inspirat?ria (r2= 0.83,R = 0.91, (95%IC 0.72 a 0.97 e p< 0.0001). Atrav?s dos resultados encontrados podemos sugerir que o manovacu?metro eletr?nico nacional ? vi?vel e seguro para realiza??o do sniff teste em sujeitos saud?veis. Em rela??o a VVM, existem diferen?as entre os g?neros na intensidade de press?o desenvolvida durante a VVM, a uma intensidade de carga durante a VVM foi considerada baixa assim como foi encontrada uma forte correla??o entre o fluxo gerado no teste e o delta de press?o expirat?rio/inspirat?rio

Avaliamos as mudanças no padrão respiratório, movimento toracoabdominal e ventilação em portadores de DPOC e indivíduos sadios. Estudou-se 45 indivíduos entre 45 e 75 anos conforme o VEF1. Utilizou-se sistemas de pletismografia respiratória por indutância, análise metabólica de gases em posição semi-sentada ao repouso e radiografia de tórax para a mobilidade diafragmática. Os grupos DPOC apresentaram redução do TI, TTOT, aumento do VC/TI, f, VE, das relações VEM/VC, VE/VO2, VE/VCO2 e diminuição da SpO2. Redução da mobilidade do diafragma e aumento da VEM/VC associaram-se à ineficiência da ventilação e a alterações no modelo ventilatório utilizado, sem alterações no movimento toracoabdominal. / We assessed changes in breathing patterns, thoracoabdominal movement and ventilation in COPD sufferers and healthy individuals. Forty-five individuals between 45 and 75 were grouped by FEV1. Inductive plethysmographic equipment, respiratory metabolism measuring (with subject at rest in semi-recumbent position), and radiographic measurement of diaphragm mobility were used. The COPD groups presented reduction in TI and TTOT and increased VT/TI, f, VE, and VD/VT, VE/VO2, VE/VCO2 and decreased SpO2. Reduction in diaphragm mobility and increase of VEM/VC were associated with ventilatory inefficiency and alterations in the ventilatory model used. No alterations in thoracoabdominal movement