Diss. (sammanfattning) Linköping : Linköpings universitet, 2005. / Härtill 5 uppsatser.

Η ΧΑΠ είναι νόσος με πολλούς φαινοτύπους, με παθοφυσιολογία που διαφέρει σε κάθε έναν από αυτούς, με κοινό χαρακτηριστικό την πτώση του λόγου FEV1/FVC. Παρόλα ταύτα πολύ πρόσφατες μελέτες δείχνουν πως δεν υπάρχει σαφής συσχέτιση μεταξύ της προσεκτικά μετρούμενης φλεγμονής στους αεραγωγούς και της πτώσης της FEV1 σε ομάδες ασθενών με ΧΑΠ, κάτι που δυνητικά μπορεί να ανατρέψει βιβλιογραφία 40 ετών [Roy K, et al 2009]. Στόχος όλων των γενετικών μελετών στη ΧΑΠ είναι ο διαχωρισμός των φαινοτύπων και η δημιουργία του γενετικού προφίλ της νόσου. Γνωρίζοντας πως η ΧΑΠ είναι πολυγονιδιακή νόσος και σε συνδυασμό με τη μεγάλη ποικιλότητα των φαινοτύπων της, οι γενετικές αλλοιώσεις φαίνεται πως οδηγούν σε διαφορετικό φαινότυπο, που τώρα ολιστικά ορίζεται ως νόσος «ΧΑΠ», αλλά σίγουρα στο εγγύς μέλλον θα διαχωριστεί σε επιμέρους ομάδες στα πλαίσια μιας πιο αποτελεσματικής και εξατομικευμένης θεραπευτικής προσέγγισης. Η παρούσα μελέτη μεταξύ άλλων, συνέβαλε στα εξής: α. Σχεδιασμός και αξιολόγηση εκκινητών και ιχνηθέτων για τον +134InsA/DelA πολυμορφισμό, με υψηλή διακριτική ικανότητα έναντι των αλληλίων 3Α και 4Α. β. Ανέδειξε την εμπλοκή των πολυμορφισμών+134InsA/DelA και G198T στην εμφάνιση αλλά και στη βαρύτητα της ΧΑΠ (όπως αυτή αξιολογείται με την FEV1), ενώ σκιαγραφήθηκε και το γενετικό προφίλ του ευαίσθητου στον καπνό του τσιγάρου καπνιστή, με λεπτομερή συσχέτιση των απλοτύπων των πολυμορφισμών της ΕΤ-1 που εμπλέκονται στη ΧΑΠ. γ. Ανέδειξε πιθανή εμπλοκή του +134InsA/DelA πολυμορφισμού στην στατική υπερδιάταση και στις αυξημένες αντιστάσεις στη ροή του αέρα στους πνεύμονες δ. Σκιαγράφησε τη σχέση των +134InsA/DelA και G198T με την ανοχή στην άσκηση και συνέκρινε τα αποτελέσματα αυτά με όσα ήδη υπάρχουν στη βιβλιογραφία. Η παρούσα μελέτη είναι η πρώτη που συσχετίζει πολυμορφισμούς με πολλαπλές αξιολογήσεις της αναπνευστικής λειτουργίας, πέραν της FEV1, κάτι που σκιαγραφεί με λεπτομέρεια το φαινότυπο της ΧΑΠ. / Chronic Obstructive Pulmonary Disease (COPD) is an entity with many phenotypes, different pathophysiological characteristics, that exhibits in all cases a diminished FEV to FVC ratio. Nevertheless, recent studies show that there is not a strict relationship between airway inflammation and FEV1 decline in patients with COPD, contrasting a 40 year literature [Roy K, et al 2009]. The aim of all recent studies dealing with genetics in COPD is the distinction of different phenotypes and the elucidation of the genetic profile of the disease. COPD is a multi-gene disorder, and knowing that it is composited by many phenotypes, one can say that, in the near future, “holistic COPD phenotype” will be unraveled in many distinguished phenotypes, leading to a personalized and patient-targeted diagnostic and therapeutic approach. The current study contributed in: a. Designing primers and probes for the +134InsA/DelA polymorphism, that could clearly distinguish both 3A and 4A alleles. b. Exhibiting that both +134InsA/DelA & G198T polymorphisms are implicated in COPD progression and severity (as defined by FEV1 values). At the same time, we managed to highlight the genetic profile of the susceptible to smoke smoker, associating haplotypes and polymorphisms of Endothelin-1 (ET-1) gene (+134InsA/DelA & G198T ) with COPD. c. Showing the implication of the +134InsA/DelA polymorphism with static lung hyperinflation and increased airway resistance. d. Revealing the association of +134InsA/DelA & G198T polymorphisms with exercise tolerance. According to our knowledge, the current study is the first in the literature showing association of ET-1 gene with lung function deterioration.

This thesis is concerned with a very common disorder of iron homeostasis: iron deficiency. The specific focus is the manner in which iron deficiency influences physiological responses to hypoxia in humans. This work is predicated on observations made over many decades in vitro and in vivo, suggesting that variations in the bioavailability of iron have important consequences for certain biological processes known to depend on oxygen availability. Three separate but related studies together form the basis for this thesis. The first two, Study A and Study B, adopt a similar approach in recruiting healthy volunteers who differ according to iron status, yielding iron-deficient and iron-replete groups in both cases. In Study A, the behaviour of the pulmonary circulation is investigated during a sustained hypoxic exposure, before and after an intravenous infusion of iron. In Study B, skeletal muscle metabolism is explored, both at the level of high-energy phosphate metabolism and the integrated physiological responses to exercise on a cycle ergometer. In the third study, Study C, a different approach is taken, recruiting patients with chronic obstructive pulmonary disease (COPD), and exploring the prevalence and associations of iron deficiency in this condition. Chapters 2 and 3 describe experiments using sustained hypoxia in a normobaric chamber, during which the pulmonary circulation is assessed non-invasively using Doppler echocardiography. These reveal augmented hypoxic pulmonary vasoconstriction (HPV) in iron-deficient individuals, who also exhibit greater sensitivity to the effects of an infusion of intravenous iron. Additionally, the way in which certain circulating mediators important for iron haemostasis change over the course of these hypoxic exposures, and how iron status influences these responses, is explored. Chapter 4 reports the findings of experiments using 31P-magnetic resonance spectroscopy and cardiopulmonary exercise testing, which demonstrate abnormal whole-body metabolism in iron-deficient individuals during large muscle-mass exercise, despite the absence of a clear defect in mitochondrial oxidative phosphorylation. Intravenous iron is found to have significant effects to alter the lactate threshold in healthy individuals, but the effects are more striking in iron-deficient individuals. Collectively, these experiments imply that iron deficiency promotes a more glycolytic phenotype. Chapter 5 explores iron deficiency in COPD, a condition in which pulmonary vascular disease, hypoxia and skeletal muscle dysfunction coexist, and examines some of the difficulties in assessing iron status in the setting of a chronic inflammatory disorder. Iron deficiency is found to be common, and unexpectedly associated with significantly more severe hypoxaemia, in patients with COPD. Possible reasons for these findings, and their clinical implications, are considered. Chapter 6 provides a summary of the main conclusions to be drawn from the studies presented in this thesis.

O aumento no número de células inflamatórias, a produção anormal de citocinas pró-inflamatórias e o desequilíbrio entre a formação de radicais livres e a capacidade antioxidante geram alterações locais e sistêmicas na doença pulmonar obstrutiva crônica (DPOC), associada com disfunção e perda da massa muscular. A reabilitação pulmonar é uma modalidade de tratamento com evidência A, cujos desfechos são medidos através da melhora da capacidade de exercício físico e qualidade de vida, mas estudos recentes demonstram uma redução no estresse oxidativo induzido pelo exercício, o que potencialmente também reduziria o dano tecidual. A análise do dano de DNA em linfócitos de sangue periférico foi utilizada como possível medida de desfecho em 13 de 39 portadores de DPOC submetidos a um programa de reabilitação pulmonar (PRP) com duração de 4 meses. Todos os pacientes foram submetidos ao teste da caminhada dos seis minutos (TC6) e ao questionário de qualidade de vida Saint George (QQVSG), sendo que 13 pacientes coletaram sangue antes e depois do PRP para análise do dano de DNA pela técnica de micronúcleos. Do total de 39 portadores de DPOC, 69,23% eram do sexo masculino com idades de 63,33 ± 8,60 anos e média de VEF1 de 1,06 ± 0,55L. Após o PRP, ocorreu aumento significativo na distância percorrida no TC6 (366,84±108,42 [pré PRP] vs. 400,76±94,55 [pós PRP], p=0,001) e melhora em todos os domínios do QQVSG (Sintomas: 47,05±21,28 [pré PRP] vs. 35,28±16,92 [pós PRP], p=0,005; Atividades: 62,84±27,07 [pré PRP] vs. 56,02±24,09 [pós PRP], p=0,038; Impacto: 33,30±18,71 [pré PRP] vs. 19,97±12,11 [pós PRP], p<0,001; Total: 49,41±21,99 [pré PRP] vs. 37,61±18,96 [pós PRP], p<0,001). Quanto à avaliação do dano genético, obteve-se uma diminuição estatisticamente significativa (p=0,014) na freqüência de micronúcleos (5,53±2,14 [pré PRP] vs. 3,07±2,13 [pós PRP] ), o que não ocorreu na análise das pontes nucleoplasmáticas e buds nucleares (1,15±0,89 [pré PRP] vs. 0,76±1,01 [pós PRP], p=0,244 e 1,69±1,43 [pré PRP] vs. 1,69±2,13 [pós PRP], p=0,804, respectivamente). A redução na freqüência de micronúcleos demonstrou que o PRP não somente melhorou a qualidade de vida e o desempenho na capacidade de exercício, mas também foi capaz de reduzir o dano de DNA. / Pulmonary rehabilitation is a treatment supported by level A evidence, and its outcomes are measured by the improvement in physical exercise capacity and quality of life. The objective of this study is to investigate if pulmonary rehabilitation reduces DNA damage in peripheral blood of patients with chronic obstructive pulmonary disease. DNA damage in peripheral blood lymphocytes was used as an outcome measure in 13 of 39 patients with chronic obstructive pulmonary disease who underwent a 4-month pulmonary rehabilitation program. All patients underwent the 6- minute walk test and answered the Saint George’s respiratory questionnaire to assess quality of life. Blood was collected from 13 patients before and after pulmonary rehabilitation program to analyze DNA damage using the micronucleus technique. After pulmonary rehabilitation program, there was a significant increase in 6- minute walk distance and improvement in all the Saint George’s respiratory questionnaire domains. The evaluation of genetic damage revealed a statistically significant decrease (p = 0.014) of micronucleus frequency. No significant differences were found in the analysis of nucleoplasmic bridges or nuclear buds. The decrease of micronucleus frequency demonstrated that PRP not only improved quality of life and performance in work capacity exercises, but also reduced DNA damage.

A hiperinsuflação dinâmica (HD) é considerada um importante contribuinte para a sensação de dispnéia e interrupção do esforço físico em pacientes portadores de doença pulmonar obstrutiva crônica (DPOC). Diversas estratégias são testadas para tentar amenizar a HD e, frequentemente, utiliza-se a capacidade inspiratória (CI) para avaliar esse efeito. Os objetivos deste estudo foram verificar a presença de HD através da pletismografia logo após a suspensão do exercício e avaliar os efeitos da pressão expiratória positiva em via aérea (EPAP) na HD em pacientes portadores de DPOC submetidos ao exercício. Foram incluídos portadores de DPOC moderada a muito grave, de ambos os sexos, considerados capazes de realizarem o teste de esforço. Todos os participantes submeteram-se à mensuração de fluxos expiratórios, volumes e capacidades pulmonares, além da análise de difusão dos gases através da pletismografia. Essas medidas foram feitas antes e após o uso do broncodilatador. A seguir utilizou-se um protocolo de exercício submáximo e nova prova de função pulmonar era realizada imediatamente após o esforço físico para avaliar a presença de hiperinsuflação, ainda sob efeito do broncodilatador. Os pacientes que apresentaram sinal de HD na pletismografia foram convidados a retornar após 48 horas para repetir o mesmo protocolo de estudo, porém com uso de máscara de EPAP durante o exercício. Os parâmetros de função pulmonar foram analisados e comparados nos diferentes momentos e entre os protocolos. A amostra foi composta inicialmente por 46 pacientes, com média de idade de 65±8,5 anos, sendo 32 (70%) do sexo masculino, 25 (54%) com doença em estágio IV. Do total, 17(37%) apresentaram HD na pletismografia realizada após o teste de exercício. Após o exercício, observou-se diferença significativa entre pacientes com e sem HD apenas nas variáveis: CI (p<0,0001), CI/CPT (p=0,001), CRF/CPT (p=0,002). O uso da EPAP durante o exercício aplicado em 17 pacientes com HD não alterou de maneira significativa a capacidade pulmonar total (CPT; p=0,64), a capacidade residual funcional (CRF; p=0,09) e o volume residual (VR; p=0,10) quando comparado aos valores obtidos após exercício sem EPAP. Entretanto na comparação da CI observou-se uma menor perda de CI (p=0,02) com o uso da máscara. Verificou-se diferença significativa na comparação da relação CI/CPT antes e após o exercício em cada protocolo, ambos apresentando uma queda do valor com o exercício. Na comparação entre protocolos observou-se diferença significativa (p=0,01), representado uma queda menor da relação CI/CPT no protocolo com EPAP. Também se observaram relações VR/CPT e CRF/CPT significativamente menores (p=0,03) após o exercício com EPAP em relação ao exercício isolado. Conclui-se que 37% dos 46 pacientes apresentaram HD, detectada através da redução da CI e da sua relação com a CPT, quando avaliados imediatamente após o teste de exercício através da pletismografia. O uso da EPAP através de máscara facial reduziu a HD em teste de exercício submáximo, observado através da redução significativa da queda da CI e da relação CI/CPT, e pela menor alteração das relações VR/CPT e CRF/CPT. / Dynamic hyperinflation (DH) contributes substantially to the sensation of dyspnea and the interruption of physical exercise in patients with chronic obstructive pulmonary disease (COPD). Several strategies have been tested to mitigate DH, and inspiratory capacity (IC) is often used to measure it. The purpose of this study was investigate the presence of DH immediately after exercise interruption using plethysmography and to evaluate the effects of expiratory positive airway pressure (EPAP) on DH of patients with COPD that underwent a exercise test. The study enrolled men and women with moderate to very severe COPD who were able to perform a exercise test. All participants underwent measurement of expiratory flows, volumes and lung capacities, and gas diffusion using plethysmography before and after the use of bronchodilators. A submaximal exercise test and repeated pulmonary function tests were conducted immediately after physical exercise to evaluate hyperinflation, still under the effect of the bronchodilator. The patients with DH according to plethysmography were invited to return 48 hours later to repeat the same protocol using an EPAP mask during exercise test. Pulmonary function parameters were analyzed and compared at the different time points and between the two tests. The sample consisted of 46 patients whose mean age was 65±8.5 years; 32 (70%) were men, and 25 (54%) had stage IV disease. Plethysmography performed after the exercise test revealed DH in 17 (37%) participants. After exercise, there was a significant difference between patients with and without DH only in IC (p<0.0001), IC/TLC (p=0.001), and FRC/TLC (p=0.002). The use of EPAP during exercise in 17 patients with DH did not significantly change total lung capacity (TLC; p=0.64), functional residual capacity (FRC; p=0.09), or residual volume (RV; p=0.10) when compared with the values obtained after exercise without EPAP. However, there was a lower loss of IC (p=0.02) in the EPAP mask group. There was a significant difference in IC/TLC before and after the exercise in each test, and both groups had a decrease in this value after exercise. The comparison between groups revealed a significant difference (p=0.01) and a smaller decrease in the IC/TLC ratio in the EPAP group. Moreover, significantly lower RV/TLC and FRC/TLC (p=0.03) were found after exercise with EPAP than after exercise alone. Of the 46 study patients, 37% developed DH, detected by a reduction in IC and in IC/TLC when evaluated immediately after exercise test using plethysmography. The use of EPAP delivered by face mask reduced DH in submaximal exercise tests, indicated by a significant reduction in IC and IC/TLC decreases and smaller changes in RV/TLC and FRC/TLC.

Introdução: A característica clínica principal da DPOC é a intolerância ao exercício físico. O mecanismo dessa limitação é complexo e multifatorial. Os principais mecanismos considerados responsáveis são a hiperinsuflação pulmonar dinâmica (HD) e disfunção muscular periférica. A hipótese principal do presente estudo é que a diminuição da massa livre de gordura (MLG) nesses pacientes, além de diretamente contribuir para redução da capacidade aeróbia, poderia contribuir indiretamente causando acentuação da HD durante o exercício. Objetivo: Investigar se a quantidade de MLG tem efeitos diretos na hiperinsuflação pulmonar dinâmica, durante o exercício em pacientes com DPOC. Métodos: 38 pacientes em estádio moderado a grave realizaram teste de exercício cardiopulmonar incremental até o limite da tolerância com medidas seriadas de capacidade inspiratória (CI). A MLG foi medida pelo teste de bioimpedância elétrica de corpo inteiro. Foram coletados também dados de função pulmonar (espirometria). Resultados: A média de idade dos pacientes foi de 66,5 ± 7,3 anos de idade, com média de VEF1 de 0,98 ± 0.05L (42 ± 15% do previsto). Valores de CI no pico do exercício (uma variável inversamente relacionada com os volumes pulmonares operacionais, ou seja, quanto maior a CI menor é hiperinflação pulmonar) foram significativamente (p <0,05) correlacionados com a CI de repouso (r = 0,78), VEF1 (r = 0,66), CVF (r = 0,56), MLG (r = 0,46) e com o índice de massa livre de gordura (IMLG) (r = 0,39). No entanto, na análise multivariada apenas o VEF1 e a CI em repouso permaneceram preditivos da CI de pico de exercício. A CI de pico foi um preditor significativo da capacidade aeróbia máxima. Conclusão: A MLG apresentou relação direta com as medidas de hiperinsuflação dinâmica durante o exercício. Contudo, a associação não se manteve quando foram feitos ajustes para indicadores de limitação do fluxo aéreo expiratório (VEF1) e hiperinflação pulmonar em repouso (CI de repouso). / Purposes: Investigate if the amount of fat free mass (FFM) has direct effects in dynamic hyperinflation during exercise in COPD patients. Methods: 38 patients with moderate to severe COPD performed treadmill incremental cardiopulmonary exercise test to the limit of tolerance with serial measurements of inspiratory capacity (IC). FFM was measured by whole-body bioelectrical impedance. Results: Patients were 66.5±7.3 years-old with mean FEV1 of 0.98±0.05L (42±15% of predicted). Peak exercise values of IC (a variable inversely related with operational lung volumes, i.e. the greater IC lower is pulmonary hyperinflation) was significantly (p<0.05) correlated with IC at rest (r=0.78), FEV1 (r=0.66), FVC (r=0.56), FFM (r=0.46) and FFM index (r=0.39). However, in multivariable analyzes only FEV1 and IC at rest remained predictive of peak IC. Peak IC was a significant predictor of peak aerobic capacity. Conclusion: FFM was directly related with measurements of dynamic hyperinflation. Nonetheless, this association disappeared when adjustments were made for indicators of expiratory airflow limitation (FEV1) and lung hyperinflation at rest (rest IC).

Theoretical foundation Chronic obstructive pulmonary disease ranks among the most serious diseases in the population. The COPD prevalence is assumed at up to 7,7%. As the name shows, it is a chronic disease characteristic with typical symptoms including particularly cough, expectoration and noticeable dyspnoea. The symptoms may get worse above their common level in the course of the disease, which we call exacerbation. The presence of unpleasant breathing problems affects different life areas, restricting the individual when performing everyday activities. The patient's psyche is affected as well, including potential incidence of anxieties or depressions. Finally, the patient's subjectively perceived quality of life is significantly worsened. Goal of the thesis The goal of this thesis is to ascertain the areas in which COPD affects the quality of life. Hypotheses H1: COPD patients feel breathing problems. H2: COPD patients feel restrictions in the sphere of common daily activities. H3: COPD patients feel restrictions in the sphere of physical activity. H4: COPD patients feel restrictions in social sphere. H5: COPD patients experience negative feelings. H6: COPD patients perceive their quality of life as bad. Methodology The research investigation was implemented based on quantitative inquiry within the grant Project No. 120/2012/S "Reflection of life quality in nursing". The data collection was performed under use of quantitative method; the research investigation took place with the help of questionnaire technology. Two questionnaires were used: St. George's Respiratory Questionare Czech (SGRQ) and WHOQOL 100 generic standardized questionnaire. The questionnaires were distributed to persons with diagnosed chronic obstructive pulmonary disease. The selection set was made up through quota selection that was observed. The quotas consisted in COPD diagnosis and the respondents' gender. The proportion of men and women was specified at 500 persons, 294 men and 206 women from that number. The research set consisted of 296 men and 207 women, i.e. 503 respondents in total. The investigation took place through addressing general practitioners, specialized outpatients' departments and chest disease wards. Results The results were processed with the help of the SASD program (statistic analysis of social data). The results can be divided into three groups. The first group includes the results characterizing the structure of the research set; the second group includes the graphically represented results, and the third group includes the results processed with the help of descriptive statistics. The last two groups include results from both questionnaires used. The results allowed us obtaining information on perception of breathing problems by the patients, on the sphere of common daily activities, on problems in the sphere of movement, on social sphere, on experiencing of negative feelings and on subjective perception of quality of life. Based on the results, the hypotheses were evaluated as follows: H1: COPD patients feel breathing problems confirmed, H2: COPD patients feel restrictions in the sphere of common daily activities - confirmed, H3: COPD patients feel restrictions in the sphere of physical activity not confirmed, H4: COPD patients feel restrictions in social sphere - not confirmed, H5: COPD patients experience negative feelings confirmed, and H6: COPD patients perceive their quality of life as bad - not confirmed. Conclusion The thesis provides comprehensive view of the spheres affecting the quality of life of patients with chronic obstructive pulmonary disease. It may be used not only for nurses but also for the whole multidisciplinary team caring for the patient. The thesis may be used also as study material for nursing students. The results of the research investigation will be further presented at conferences and published in professional journals.

Orientador: Prof. Dr. Sergio Daishi Sasaki / Dissertação (mestrado) - Universidade Federal do ABC, Programa de Pós-Graduação em Biossistemas, 2015. / Embora a doença pulmonar obstrutiva crônica (DPOC)seja uma das maiores causas de morbidade e mortalidade no mundo, ainda não há tratamento que reverta àdestruição do parênquima pulmonarobservadas nestes pacientes. O desenvolvimento de modelos experimentais,que mimetizam as características e os mecanismos fisiopatológicos envolvidos no enfisema,por meioda instilação de uma protease,éde extrema relevância uma vez que podem contribuirpara o desenvolvimento de futuras estratégias terapêuticas. Uma das causas atribuídas para o aparecimento da doença éo tabagismo, no entanto, fatores genéticose fatoresambientais como a poluição também estão envolvidos com o aparecimento da doença em humanos. Algumas enzimas estão envolvidas no processo de instalação da doença como as metaloproteases 9 e 12 e as serinoproteases, especificamente a elastase de neutrófilos humana. Em um projeto anterior,utilizando o inibidor de serinoproteases recombinante rBmTI-Ano modelo de indução deenfisema pulmonar em camundongos,foi observado que esta molécula é capaz de interferir na progressão do enfisema pulmonar, o rBmTI-A é um inibidor pertencente à família Kunitz-BPTI e possui dois domínios inibitórios com atividade sobre tripsina, calicreína plasmática humana e elastase de neutrófilos humana. Neste projeto,foi utilizadooinibidor rBmTI-6,também pertencente à família Kunitz-BPTI,porém contendo três domínios inibitórios com atividade sobre tripsina bovinae plasmina. Uma amostra de rBmTI-6 previamente purificada por cromatografia de afinidadeem tripsina-sepharosefoiaplicada emuma cromatografia de troca iônica, coluna Resource Q,que separou osdomínios inibitóriosem domínio 1 (rBmTI-6-d1)e domínios 2e 3 (rBmTI-6-d2/3). O rBmTI-6-d1teve suaconcentraçãoativa determinada em 1,5µMe a constante de inibição (Ki)sobre tripsina bovina determinado em 1,95nM, este domínio purificado apresentou 77% de atividade específica (inibidor ativo em relação à proteína total da amostra). O rBmTI-6-d1 foi utilizado em um experimentono modelo experimental de enfisema pulmonar.Neste experimento o enfisema foi induzido utilizando a instilação por via nasal de elastase pancreática porcina(PPE)e 1 hora depois da instilaçãocom PPEfoi feito o tratamento coma instilaçãorBmTI-6-d1. Vinte e um dias após a instilação de PPEe rBmTI-6-d1 foi feita a analise da função respiratória para os14 parâmetros de elastância tecidual (Htis),resistência tecidual (Gtis)eresistência das vias aéreas (Raw). Animais induzidos ao enfisema e tratados com o rBmTI-6-d1apresentaram valores de elastância tecidual (Htis)e resistência tecidual (Gtis) próximasaosvalores dosgrupos controles, cujos animais não foram induzidos ao enfisema, os animais não tratados com rBmTI-6-d1 apresentaram aumento de Htis e Gtis. Para o parâmetro de resistência das vias aéreas o resultado mostra uma tendência de redução deste parâmetro para animais tratados com rBmTI-6-d1. Os resultados da medida do intercepto linear médio (Lm), apontam uma tendência ao grupo que foi induzido ao enfisema e tratado com rBmTI-6-d1 de redução da Lm em comparação com o grupo induzido ao enfisema e não tratado.Estes resultados sugerem que o inibidor teve papel de prevenir a instalação do enfisema pulmonar nos animais tratados com rBmTI-6-d1. / Although chronic obstructive pulmonary disease (COPD) is a major cause of morbidity and mortality worldwide, there is no treatment to reverse the destruction of the lung parenchymaobserved in these patients. The development of experimental models that mimic the characteristics and pathophysiological mechanisms of emphysema by instillationone protease is extremely important since it can contribute to the development of future therapeutic strategies. One of the causes associated with the onset of the disease is smoking, however, genetic factors and environmental factors such as pollutionare also involved in disease in humans. Some enzymes are involved in the installation process of the disease like metalloproteases 9 and 12 and serineproteases, specifically human neutrophil elastase. In a previous study using recombinant serineprotease inhibitor rBmTI-A in a pulmonary emphysema model with mice was observed that this molecule is capable of interfering with the progression of the disease, rBmTI-A is an inhibitor belongingto the Kunitz-BPTI family and has two inhibitory domains with activity towardtrypsin, human plasmakallikreinand human neutrophil elastase. In this project, a sample of rBmTI-6 inhibitor also belongs to the Kunitz-BPTI family, previously purified by affinity chromatography on trypsin-sepharose was used, the sample was purified again using an ion exchange chromatography with Resource Q column, where it was observed a separation of the inhibitory domains. The domain 1 (rBmTI-6-d1) had their active concentration determined in 1,5?M. The rBmTI-6-d1 had a Ki on certain bovine trypsin in 1,95nM this purified domain had 77% specific activity (active inhibitor in the total protein of the sample). The rBmTI-6-d1 was selectedfor the experiments with an experimental model of pulmonary emphysema. In these experiments emphysema was induced by intranasal instillation of porcine pancreatic elastase (PPE) and 1 hour after PPE instillation, the treatment with rBmTI-6-d1. Twenty-one days after instillation of PPE and rBmTI-6-d1 was made the analysis of respiratory function to tissue elastance parameters (Htis), tissue resistance (Gtis) and airway resistance (Raw). Animals induced emphysema and treated with rBmTI-6-d1 showedtissue elastance values (Htis) and tissue resistance (Gtis) close to the values of the control groups, whose animals were not induced emphysema, untreated animals with rBmTI-6-d1 increased by Htis and Gtis. For the air way resistance the result shows a tendency to decrease in this parameter for animals treated with rBmTI-6-d1. The results of measuring the mean linear intercept (Lm), show a tendency to emphysema group that was induced and treated with -rBmTI6-d1 Lm reduced in comparison with the experimental group and the untreated emphysema. These results suggest that the inhibitory role was to prevent the development of pulmonary emphysema in animals treated with rBmTI6-d1.

Made available in DSpace on 2014-06-11T19:32:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-03-16Bitstream added on 2014-06-13T20:42:56Z : No. of bitstreams: 1 faganello_mm_dr_botfm.pdf: 465819 bytes, checksum: 7899369510ab22be938ad270a777ebd2 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Nos últimos anos, vários estudos avaliaram os marcadores da doença associados à freqüência de exacerbação, hospitalização, readmissão e mortalidade em pacientes portadores de doença pulmonar obstrutiva crônica (DPOC). Entretanto, estudos que avaliaram os marcadores associados à ocorrência de exacerbação em pacientes ambulatoriais são limitados. Assim, o objetivo deste estudo foi identificar os fatores de predição da ocorrência de exacerbação no período de um ano em 120 pacientes com DPOC atendidos no Ambulatório de Pneumologia da Faculdade de Medicina de Botucatu - Unesp. Os pacientes tiveram o diagnóstico de DPOC confirmado e foram submetidos às seguintes avaliações: espirometria pré e pós-broncodilatador, composição do corpo (antropometria e bioimpedância), qualidade de vida por meio do Saint George s Respiratory Questionnaire (SGRQ), intensidade da dispnéia por meio da escala modificada Medical Research Council (MMRC) e do índice basal de dispnéia (BDI) e tolerância ao exercício (distância percorrida em 6 minutos DP6). Em seguida foi calculado o índice BODE de acordo com os pontos de corte do volume expiratório no primeiro segundo (VEF1), do índice de massa do corpo, do MMRC e da DP6. Durante o período de acompanhamento de um ano, 60 pacientes (50%) apresentaram pelo menos um episódio de exacerbação da doença e, em conseqüência da agudização, 25 pacientes foram hospitalizados. Comorbidades extra-pulmonares foram causa de hospitalização em oito pacientes e de óbito em cinco pacientes. Na avaliação inicial, os pacientes que exacerbaram tinham maior comprometimento da função pulmonar e da troca gasosa, valores mais elevados do índice BODE e maior proporção de pacientes com DPOC III e IV. Além disso, apresentavam menores valores de DP6, maior sensação de dispnéi... / Markers of disease severity have been associated with mortality, occurrence and frequency of hospitalization and readmission due disease exacerbation in chronic obstructive pulmonary disease (COPD) patients. However, information about predictor factors for the occurrence of exacerbation in ambulatory COPD patients is scarce. Therefore, the aim of the present study was to identify predictor factors for the occurrence of exacerbation in 120 patients with COPD followed during one year in the outpatient clinic at Universidade Estadual Paulista (UNESP, Paulista State University) School of Medicine at Botucatu, located in the State of São Paulo, Brazil. The patients had the diagnosis of COPD confirmed and underwent to the following evaluations: pre- and postbronchodilator spirometry, body composition (anthropometry and bioimpedance), health-related quality of life (Saint George s Respiratory Questionnaire -SGRQ), dyspnea scores (Medical Research Council MMRC and basal dispnea index -BDI) and exercise tolerance (6MWD). Bode index was calculated taking in consideration the cutt off points for forced volume in the first second (FEV1), body mass index, MMRC and 6MWD. During the followup period 60 patients (50%) presented at least one exacerbation episode and, as consequence, 25 patients were hospitalized. Eight patients were hospitalized and five died due to non-pulmonary comorbidities. At baseline, patients with exacerbations during the follow-up period presented lower values of airway obstruction indexes and of arterial blood gases and higher values of BODE score and proportion of COPD patients class III and IV. In addition, the values of 6MWD were lower, dyspnea sensation was higher and the health- related quality of life was more deteriorated in these patients. No significative associations were found between gender, corticosteroid use... (Complete abstract click electronic access below)

O aumento no número de células inflamatórias, a produção anormal de citocinas pró-inflamatórias e o desequilíbrio entre a formação de radicais livres e a capacidade antioxidante geram alterações locais e sistêmicas na doença pulmonar obstrutiva crônica (DPOC), associada com disfunção e perda da massa muscular. A reabilitação pulmonar é uma modalidade de tratamento com evidência A, cujos desfechos são medidos através da melhora da capacidade de exercício físico e qualidade de vida, mas estudos recentes demonstram uma redução no estresse oxidativo induzido pelo exercício, o que potencialmente também reduziria o dano tecidual. A análise do dano de DNA em linfócitos de sangue periférico foi utilizada como possível medida de desfecho em 13 de 39 portadores de DPOC submetidos a um programa de reabilitação pulmonar (PRP) com duração de 4 meses. Todos os pacientes foram submetidos ao teste da caminhada dos seis minutos (TC6) e ao questionário de qualidade de vida Saint George (QQVSG), sendo que 13 pacientes coletaram sangue antes e depois do PRP para análise do dano de DNA pela técnica de micronúcleos. Do total de 39 portadores de DPOC, 69,23% eram do sexo masculino com idades de 63,33 ± 8,60 anos e média de VEF1 de 1,06 ± 0,55L. Após o PRP, ocorreu aumento significativo na distância percorrida no TC6 (366,84±108,42 [pré PRP] vs. 400,76±94,55 [pós PRP], p=0,001) e melhora em todos os domínios do QQVSG (Sintomas: 47,05±21,28 [pré PRP] vs. 35,28±16,92 [pós PRP], p=0,005; Atividades: 62,84±27,07 [pré PRP] vs. 56,02±24,09 [pós PRP], p=0,038; Impacto: 33,30±18,71 [pré PRP] vs. 19,97±12,11 [pós PRP], p<0,001; Total: 49,41±21,99 [pré PRP] vs. 37,61±18,96 [pós PRP], p<0,001). Quanto à avaliação do dano genético, obteve-se uma diminuição estatisticamente significativa (p=0,014) na freqüência de micronúcleos (5,53±2,14 [pré PRP] vs. 3,07±2,13 [pós PRP] ), o que não ocorreu na análise das pontes nucleoplasmáticas e buds nucleares (1,15±0,89 [pré PRP] vs. 0,76±1,01 [pós PRP], p=0,244 e 1,69±1,43 [pré PRP] vs. 1,69±2,13 [pós PRP], p=0,804, respectivamente). A redução na freqüência de micronúcleos demonstrou que o PRP não somente melhorou a qualidade de vida e o desempenho na capacidade de exercício, mas também foi capaz de reduzir o dano de DNA. / Pulmonary rehabilitation is a treatment supported by level A evidence, and its outcomes are measured by the improvement in physical exercise capacity and quality of life. The objective of this study is to investigate if pulmonary rehabilitation reduces DNA damage in peripheral blood of patients with chronic obstructive pulmonary disease. DNA damage in peripheral blood lymphocytes was used as an outcome measure in 13 of 39 patients with chronic obstructive pulmonary disease who underwent a 4-month pulmonary rehabilitation program. All patients underwent the 6- minute walk test and answered the Saint George’s respiratory questionnaire to assess quality of life. Blood was collected from 13 patients before and after pulmonary rehabilitation program to analyze DNA damage using the micronucleus technique. After pulmonary rehabilitation program, there was a significant increase in 6- minute walk distance and improvement in all the Saint George’s respiratory questionnaire domains. The evaluation of genetic damage revealed a statistically significant decrease (p = 0.014) of micronucleus frequency. No significant differences were found in the analysis of nucleoplasmic bridges or nuclear buds. The decrease of micronucleus frequency demonstrated that PRP not only improved quality of life and performance in work capacity exercises, but also reduced DNA damage.