Tsang Wing Tai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 132-157). / Abstracts in English and Chinese. / Contents --- p.i / Abstract - English --- p.iv / Abstract - Chinese --- p.vi / Statement of Originality --- p.viii / Acknowledgements --- p.ix / List of Figures --- p.x / List of Tables --- p.xii / Abbreviations --- p.xiii / Chapter Chapter 1: --- Objectives and planning of studies in this thesis --- p.1 / Chapter Chapter 2: --- Literature Reviews / Chapter 2.1 --- Definition of asthma --- p.2 / Chapter 2.2 --- Diagnosis and classification of asthma in children --- p.5 / Chapter 2.3 --- Assessment of asthma in school-age children --- p.7 / Chapter 2.4 --- The use of HSCSI in clinical trial --- p.27 / Chapter 2.5 --- Problems of sputum induction in children --- p.32 / Chapter 2.6 --- Conclusion --- p.34 / Chapter Chapter 3: --- HSCSI - Methodology and Materials / Chapter 3.1 --- Introduction --- p.35 / Chapter 3.2 --- HSCSI - Methodology --- p.37 / Chapter 3.3 --- Protocol for combined HSCSI --- p.45 / Chapter 3.4 --- Practical considerations in HSCSI --- p.49 / Chapter 3.5 --- Expression of airway response --- p.50 / Chapter 3.6 --- Safety issue in HSCSI --- p.52 / Chapter 3.7 --- Approach in our studies --- p.54 / Chapter 3.8 --- Sputum processing --- p.55 / Chapter 3.9 --- Interpretation of sputum differential cell counts --- p.68 / Chapter 3.10 --- Conclusion --- p.73 / Chapter Chapter 4: --- Factors predicting successful sputum induction / Chapter 4.1 --- Introduction --- p.74 / Chapter 4.2 --- Methods --- p.76 / Chapter 4.3 --- Results --- p.80 / Chapter 4.4 --- Discussion --- p.82 / Chapter 4.5 --- Conclusion --- p.88 / Chapter Chapter 5: --- Use of once-daily fluticasone propionate in children with stable asthma -Study on airway inflammatory markers / Chapter 5.1 --- Introduction --- p.89 / Chapter 5.2 --- Methods --- p.92 / Chapter 5.3 --- Results --- p.97 / Chapter 5.4 --- Discussion --- p.101 / Chapter 5.5 --- Conclusion --- p.105 / Chapter Chapter 6 --- Assessment of cough frequency in children with stable asthma -Study on airway inflammatory markers / Chapter 6.1 --- Introduction --- p.106 / Chapter 6.2 --- Cough Monitoring Machine LR 102 --- p.109 / Chapter 6.3 --- Methods --- p.114 / Chapter 6.4 --- Results --- p.119 / Chapter 6.5 --- Discussion --- p.123 / Chapter 6.6 --- Conclusions --- p.127 / Chapter Chapter 7 --- Overall summary and conclusion --- p.128 / References --- p.132 / Appendix I Assessment form 226}0ؤ HSCSI

Since the discovery of cyclosporine in 1971, calcineurin inhibitors have played a critical role in the therapeutic suppression of the immune response. Patients receiving solid organ transplants rely heavily on these medications to prevent the acute and chronic rejection of allografted tissue. Introduction of tacrolimus, the most frequently prescribed calcineurin inhibitor, has lead to improved clinical outcomes for organ transplant recipients; however, little improvement has been noted in lung transplantation. Difficulties with current oral dosing regimens for lung transplant patients stem primarily from drug systemic toxicity, heightened risk of invasive infection, and erratic oral bioavailability. We have proposed that pulmonary delivery of a tacrolimus formulation with improved solubility can provide high lung concentrations, while limiting corresponding systemic levels associated with toxicity. Chapter 2 investigates the pulmonary administration of tacrolimus dispersion for nebulization to lung transplanted rats. Resulting lung and blood levels were determined by appropriate bioanalytical methods. Limited systemic absorption was seen after pulmonary delivery, resulting in a 50 to 1 lung to blood concentration ratio. A 28 day safety and stability evaluation of tacrolimus dispersion for nebulization was conducted in Chapter 3. Results showed no signs of toxicity in Sprague Dawley rats and proved the stability of tacrolimus powder for dispersion for 3 months. For cases of severe asthma, immunosuppression is also necessary to restore normal lungs function and is typically treated with corticosteroids. Corticosteroids, however, are well known for their untoward side effects and can prove ineffective in severe asthmatics that have developed corticosteroid resistance. Chapter 4 investigates the use of tacrolimus dispersion for nebulization for prophylactic treatment of asthma. Efficacy was determined in an asthma-induced animal model by quantification of inflammatory cells and signaling chemicals. In Chapter 5, tacrolimus powder for inhalation is investigated in a novel dry powder inhalation platform. Respirable particles are produced when bulk particles (500 [micrometer]) comprising a matrix of drug/excipient are sheared apart by a marketed inhalation device to produce particles of the appropriate geometric diameter (50 [micrometer]). Biocompatible material with brittle properties were found to produce fine particle fractions (FPF) up to 70.3% and total emitted doses (TED) higher than 95%. / text

Lau, Ka Hoi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 139-151). / Abstracts in English and Chinese. / DECLARATION --- p.i / ACKNOWLEDGEMENT --- p.ii / ABBREVIATIONS --- p.iii / ABSTRACT IN ENGLISH --- p.iv / ABSTRACT IN CHINESE --- p.vii / TABLE OF CONTENTS --- p.x / Chapter CHAPTER I - --- INTRODUCTION / Chapter 1.1 --- Regulation of human airway surface liquid --- p.1 / Chapter 1.2 --- Cysteinyl leukotrienes in asthma --- p.2 / Chapter 1.3 --- Cysteinyl leukotriene receptor in epithelial cells --- p.5 / Chapter 1.4 --- Particular interest on CysLT1 receptor --- p.7 / Chapter 1.5 --- Cysteinyl leukotrienes receptor antagonists --- p.10 / Chapter 1.6 --- Purinergic receptors in epithelial cells --- p.11 / Chapter 1.7 --- P2Y receptors in epithelial cells --- p.13 / Chapter 1.8 --- Signalling pathways of P2Y receptors by nucleotide stimulation --- p.15 / Chapter 1.9 --- The importance of P2Y6 receptor on inflammation --- p.17 / Chapter 1.10 --- Relation between CysLT1 receptor and P2Y receptor --- p.18 / Chapter 1.11 --- The properties of 16HBE14o- cell line --- p.21 / Chapter 1.12 --- Objectives of the present project --- p.22 / Chapter CHAPTER II - --- MATERIALS AND METHODS / Chapter 2.1 --- Solutions and chemicals --- p.23 / Chapter 2.2 --- Cell culture --- p.25 / Chapter 2.3 --- Measurement of intracellular calcium concentration ([Ca2+ ]i) with fluorescent imaging / Chapter 2.3.1 --- Preparation of 16HBE14o- cells for fluorescent imaging --- p.26 / Chapter 2.3.2 --- Measurement of [Ca2+]j with fluorescent imaging --- p.28 / Chapter 2.4 --- Measurement of short-circuit current (Isc) and transepithelial resistance with Ussing chamber / Chapter 2.4.1 --- Preparation of 16HBE14o- cells for Isc and transepithelial resistance measurement --- p.31 / Chapter 2.4.2 --- Measurement of Isc and transepithelial resistance with Ussing chamber --- p.33 / Chapter 2.5 --- Immunoblot analysis for CysLT1 and P2Y6 receptors --- p.35 / Chapter 2.6 --- Measurement of protein kinase A activity --- p.36 / Chapter 2.7 --- Data analysis --- p.37 / Chapter CHAPTER III - --- RESULTS / Chapter 3.1 --- Expressions of CysLTi and P2Y6 receptor in 16HBE14o- cell monolayers --- p.38 / Chapter 3.2 --- "Differential inhibitory effects of montelukast, pranlukast and zafirlukast to UDP on Isc and [Ca2+]i in 16HBE14o- cells" / Chapter 3.2.1 --- Effect of apical or basolateral application of UDP on Isc and [Ca2+]i --- p.41 / Chapter 3.2.2 --- Effect of montelukast to the application of UDP on Isc and [Ca2+]i --- p.48 / Chapter 3.2.3 --- Effect of pranlukast to the application of UDP on Isc and [Ca2+ ]i --- p.57 / Chapter 3.2.4 --- Effect of zafirlukast to the application of UDP on Isc and [Ca2+]j --- p.63 / Chapter 3.2.5 --- "Summary of the effects of montelukast, pranlukast, zafirlukast to UDP application on Isc and [Ca2+]i" --- p.69 / Chapter 3.3 --- Cellular mechanism(s) underlying the effect of montelukast to apical UDP application on 16HBE14o-cells / Chapter 3.3.1 --- Effect of various blockers inhibiting Ca2 226}Bؤdependent pathway on UDP-induced [Ca2+]i in the presence or absence of montelukast --- p.70 / Chapter 3.3.2 --- "Effects of montelukast, pranlukast and zafirlukast to PKA or Epac on Isc induced by apical UDP" --- p.86 / Chapter 3.4 --- "Effects of montelukast, pranlukast and zafirlukast on other P2Y receptor agonists on 16HBE14o- cells" / Chapter 3.4.1 --- "Effects of montelukast, pranlukast and zafirlukast on 2-methio-ADP-induced Isc and [Ca2+]i responses on 16HBE14o- cellsl" --- p.14 / Chapter 3.4.2 --- "Effects of montelukast, pranlukast and zafirlukast on UTP-induced Isc and [Ca2+]i responses on 16HBE14o- cells" --- p.116 / Chapter CHAPTER IV - --- DISCUSSION / Chapter 4.1 --- Differential effects of CysLT1 antagonists to P2Y6 agonist on Isc and [Ca2+]i in 16HBE14o-cells --- p.120 / Chapter 4.2 --- Possible cellular mechanism(s) underlying the effects of CysLT1 antagonists on UDP-induced [Ca2+]j increase in 16HBE14o- cells --- p.125 / Chapter 4.3 --- Possible cellular mechanism(s) underlying the effects of CysLT1 antagonists on UDP-induced Isc in 16HBE14o- cells --- p.129 / Chapter 4.4 --- Effects of CysLT1antagonists on other P2Y receptor subtypes in 16HBE14o- cells --- p.132 / Chapter 4.5 --- Summary: Possible interaction between CysLT1 antagonists and P2Y6 receptor --- p.135 / Chapter 4.6 --- Clinical implications and perspectives --- p.138 / Chapter CHAPTER V - --- REFERENCES --- p.139

Introdução: A asma é caracterizada por um processo inflamatório crônico que está associado ao desenvolvimento da hiperresponsividade brônquica (HRB). O exercício físico regular proporciona inúmeros benefícios aos pacientes com asma porém, os efeitos do treinamento físico na HRB permanecem pouco compreendidos. Objetivo: Avaliar o efeito do treinamento físico aeróbio na hiperresponsividade brônquica, inflamação pulmonar, controle clínico e fatores relacionados à qualidade de vida de pacientes adultos com asma persistente moderada a grave. Métodos: Cinquenta e oito adultos com asma moderada a grave foram divididos aleatoriamente, em dois grupos: Controle (GC, n=28) e Treinado (GT, n=30). Os pacientes do GC foram submetidos a um programa educacional e a um programa de exercícios respiratórios, enquanto os pacientes do GT foram submetidos a todos os procedimentos do GC e a um programa de condicionamento físico aeróbio. A hiperresponsividade brônquica foi avaliada através do teste de broncoprovocação inespecífica com histamina antes e após a intervenção. Nestas ocasiões, todos os pacientes também realizaram, análise do escarro induzido e da fração exalada de óxido nítrico, espirometria, teste ergoespirométrico e responderam aos questionários de controle clínico, fatores de saúde relacionados à qualidade de vida (FSRQV) e níveis de depressão. Além disso, foi coletada uma amostra do sangue venoso dos pacientes para quantificação do IgE total e de IgE específica. Resultados: Após três meses de intervenção, os pacientes do GT aumentaram 1 dupla dose de concentração (dd) (1 dd; 0,3-1,7 dd, 95% CI) (p < 0,05) enquanto o GC (0,06 dd; -0,6dd a 0,7 dd, 95% CI) não apresentou mudança significativa na hiperresponsividade brônquica. A inflamação pulmonar reduziu apenas nos pacientes do GT que apresentaram níveis elevados de eosinófilos (> 3%) e FeNO (> 26ppb) (p < 0,05). O condicionamento aeróbio melhorou os FSRQV, controle clínico da asma e níveis de depressão (p < 0,05). Conclusão: Nossos resultados demonstram que o treinamento aeróbio tem um efeito anti-inflamatório importante na asma e deve ser considerado como um tratamento complementar para o manejo da doença / Introduction: Asthma is characterized by a chronic inflammatory process that is associated with the development of bronchial hyperresponsiveness (BHR). Regular exercise provides numerous benefits in patients with asthma; however, the effects of exercise training on BHR remain poorly understood. Objective: To evaluate the effect an aerobic training on bronchial hyperresponsiveness, pulmonary inflammation, clinical control and health related quality of life (HRQoL) in adults patients with moderate to severe asthma. Methods: Fifty-eigth patients adults with moderate to severe asthma were randomly assigned into two groups: Control (CG, n = 28) and Trained (TG, n = 30).The GC patients undertake an educational program and performed breathing exercises, while the TG patients underwent the same procedures than CG plus an aerobic training program. Bronchial hyperresponsiveness was assessed by nonspecific bronchial provocation test with histamine before and after the intervention. On these occasions, all patients also performed induced sputum analysis and fractional exhaled nitric oxide (FeNO), spirometry, cardiopulmonary exercise testing and fulfilled questionnaires to evaluate clinical control test, HRQoL and depression levels. In addition, blood samples were collect in order to quantify total serum immunoglobulin (IgE) and specific IgE. Results: After 3 months of intervention, the TG increased 1 double dose of concentration (dd) (0.3 to 1.7 dd, 95% IC) and CG did not change significantly on bronchial hyperresponsiveness 0.06 dd (-0.6 to 0.7 dd, 95% IC) (p < 0.05).The pulmonary inflammation reduced only in the GT patients with high levels of eosinophils (> 3%) and FeNO (> 26ppb) (p < 0.05). Aerobic training also improved HRQoL, clinical control and depression levels (p < 0.05).Conclusion: Our results demonstrate that aerobic training exercise has a significant anti-inflammatory effect on asthma and should be considered as a complementary treatment for disease management

Introdução: A asma é caracterizada por um processo inflamatório crônico que está associado ao desenvolvimento da hiperresponsividade brônquica (HRB). O exercício físico regular proporciona inúmeros benefícios aos pacientes com asma porém, os efeitos do treinamento físico na HRB permanecem pouco compreendidos. Objetivo: Avaliar o efeito do treinamento físico aeróbio na hiperresponsividade brônquica, inflamação pulmonar, controle clínico e fatores relacionados à qualidade de vida de pacientes adultos com asma persistente moderada a grave. Métodos: Cinquenta e oito adultos com asma moderada a grave foram divididos aleatoriamente, em dois grupos: Controle (GC, n=28) e Treinado (GT, n=30). Os pacientes do GC foram submetidos a um programa educacional e a um programa de exercícios respiratórios, enquanto os pacientes do GT foram submetidos a todos os procedimentos do GC e a um programa de condicionamento físico aeróbio. A hiperresponsividade brônquica foi avaliada através do teste de broncoprovocação inespecífica com histamina antes e após a intervenção. Nestas ocasiões, todos os pacientes também realizaram, análise do escarro induzido e da fração exalada de óxido nítrico, espirometria, teste ergoespirométrico e responderam aos questionários de controle clínico, fatores de saúde relacionados à qualidade de vida (FSRQV) e níveis de depressão. Além disso, foi coletada uma amostra do sangue venoso dos pacientes para quantificação do IgE total e de IgE específica. Resultados: Após três meses de intervenção, os pacientes do GT aumentaram 1 dupla dose de concentração (dd) (1 dd; 0,3-1,7 dd, 95% CI) (p < 0,05) enquanto o GC (0,06 dd; -0,6dd a 0,7 dd, 95% CI) não apresentou mudança significativa na hiperresponsividade brônquica. A inflamação pulmonar reduziu apenas nos pacientes do GT que apresentaram níveis elevados de eosinófilos (> 3%) e FeNO (> 26ppb) (p < 0,05). O condicionamento aeróbio melhorou os FSRQV, controle clínico da asma e níveis de depressão (p < 0,05). Conclusão: Nossos resultados demonstram que o treinamento aeróbio tem um efeito anti-inflamatório importante na asma e deve ser considerado como um tratamento complementar para o manejo da doença / Introduction: Asthma is characterized by a chronic inflammatory process that is associated with the development of bronchial hyperresponsiveness (BHR). Regular exercise provides numerous benefits in patients with asthma; however, the effects of exercise training on BHR remain poorly understood. Objective: To evaluate the effect an aerobic training on bronchial hyperresponsiveness, pulmonary inflammation, clinical control and health related quality of life (HRQoL) in adults patients with moderate to severe asthma. Methods: Fifty-eigth patients adults with moderate to severe asthma were randomly assigned into two groups: Control (CG, n = 28) and Trained (TG, n = 30).The GC patients undertake an educational program and performed breathing exercises, while the TG patients underwent the same procedures than CG plus an aerobic training program. Bronchial hyperresponsiveness was assessed by nonspecific bronchial provocation test with histamine before and after the intervention. On these occasions, all patients also performed induced sputum analysis and fractional exhaled nitric oxide (FeNO), spirometry, cardiopulmonary exercise testing and fulfilled questionnaires to evaluate clinical control test, HRQoL and depression levels. In addition, blood samples were collect in order to quantify total serum immunoglobulin (IgE) and specific IgE. Results: After 3 months of intervention, the TG increased 1 double dose of concentration (dd) (0.3 to 1.7 dd, 95% IC) and CG did not change significantly on bronchial hyperresponsiveness 0.06 dd (-0.6 to 0.7 dd, 95% IC) (p < 0.05).The pulmonary inflammation reduced only in the GT patients with high levels of eosinophils (> 3%) and FeNO (> 26ppb) (p < 0.05). Aerobic training also improved HRQoL, clinical control and depression levels (p < 0.05).Conclusion: Our results demonstrate that aerobic training exercise has a significant anti-inflammatory effect on asthma and should be considered as a complementary treatment for disease management