Indiana University-Purdue University Indianapolis (IUPUI) / Rationale: Tissue remodeling and complement activation are asthma hallmarks. Type V collagen [col(V)], a cryptic antigen, becomes exposed during lung remodeling. IL-17 is key to anti-col(V) immunity, and regulates complement activation. We have reported that col(V)-induced tolerance down regulates IL-17 and prevents immune-mediated lung diseases. Objectives: Determine a role for anti-col(V) immunity in asthma. Methods: Serum anti-col(V) antibodies were measured in asthma patients, and immunohistochemistry utilized to detect interstitial col(V) in fatal asthma. Balb/c mice were tolerized with col(V) prior to sensitization with ovalbumin (OVA), and subsequent OVA intranasal challenge. Airway hyper-responsiveness (AHR) to methacholine was measured; and RT-PCR utilized to determine local Il17 transcripts. Bronchoalveolar lavage levels of C3a¸ C5a and OVA-specific IgE were measured; and immunohistochemistry utilized to detect expression of complement regulatory proteins, expression, CD46/Crry and CD55, in lung tissue. Results: Compared to normal subjects, anti-col(V) antibodies were increased in asthmatics; and interstitial col(V) was over expressed in fatal asthma. OVA-induced AHR up regulated anti-col(V) antibodies systemically, and increased OVA-specific IgE and C3a in BAL, and parenchymal Il17 transcripts. Col(V)-induced tolerance abrogated AHR, down regulated OVA-induced T cell proliferation, as well as total and OVA-specific IgE, C3a, IL-17 expression and tracheal smooth muscle contraction. Crry/CD46 and CD55, key to preventing complement activation, were down regulated on goblet cells in murine allergic airway disease. Conclusions: Anti-col(V) immunity correlates with asthma pathogenesis, and col(V)-induced tolerance may be a novel therapeutic for asthma. Decreased expression of Crry/CD46 and CD55 on goblet cells may in part account for complement activation in asthma.

He, Qiqiang. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 142-154). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese.

Lau Yin Kei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 58-79). / Abstracts in English and Chinese. / Acknowledgement --- p.I / Abstract --- p.IV / Abstract in Chinese --- p.VI / Abbreviations --- p.VIII / Introduction --- p.1 / Chapter 1.1 --- Prevalence of COPD and asthma in Hong Kong --- p.1 / Chapter 1.2 --- Players in pathogenesis of COPD --- p.2 / Chapter 1.3 --- Players in pathogenesis of asthma --- p.4 / Chapter 1.4 --- The use of exhaled breath condensate in previous studies --- p.6 / Chapter 1. 5 --- Brief overview of chemokines --- p.8 / Chapter 1.6 --- Objective of this study --- p.12 / Materials and methods --- p.14 / Chapter 2.1 --- Study population --- p.14 / Chapter 2.1.1 --- Patients with COPD and control subjects --- p.14 / Chapter 2.1.2 --- Patients with asthma and control subjects --- p.15 / Chapter 2.2 --- Lung function --- p.15 / Chapter 2.3 --- Dyspnoea score measurement of patients with COPD --- p.16 / Chapter 2.4 --- Classification of patients and asthma severity --- p.16 / Chapter 2.5 --- Skin prick test and blood tests --- p.16 / Chapter 2.6 --- Collection of exhaled breath condensate --- p.17 / Chapter 2.7 --- Measurement of constituent in EBC --- p.17 / Chapter 2.7.1 --- "Measurement of 8-isoprostane, MCP-1 and GROα in patients with COPD and the corresponding control subjects" --- p.17 / Chapter 2.7.2 --- Measurement of eotaxin and MDC of patients with asthma and the corresponding control subjects --- p.18 / Chapter 2.8 --- Reproducibility of exhaled breath constituent --- p.18 / Chapter 2.8.1 --- "Assessment of reproducibility of the exhaled MCP-1, GROα and8- isoprostane measurements" --- p.19 / Chapter 2.8.2 --- Assessment of reproducibility of the exhaled eotaxin and MDC measurement --- p.19 / Chapter 2.9 --- Statistical analysis --- p.19 / Results --- p.21 / Chapter 3.1 --- Patients with COPD and corresponding control subjects --- p.21 / Chapter 3.2 --- Patients with asthma and corresponding control subjects --- p.28 / Discussion --- p.36 / Chapter 4.1 --- "Exhaled 8-isoprostane, GRO-α and MCP-1 of patients with COPD and corresponding control subjects" --- p.36 / Chapter 4.2 --- Exhaled eotaxin and MDC from patients with asthma and corresponding control subjects --- p.43 / Chapter 4.3 --- Technical aspects of EBC assessment --- p.49 / Future prospect --- p.54 / Conclusion --- p.56 / References --- p.58 / Tables and Figures / Table 1. Demographics of the COPD and control subjects --- p.22 / Figure 1. The level of 8-isoprostane in the exhaled breath condensate of COPD and control subjects --- p.23 / Figure 2. The level of GROa in the exhaled breath condensate of COPD and control subjects --- p.25 / "Figure 3 Bland and Altman's Plot of the repeatability of 8-isoprostane, GROa and MCP-1 in the exhaled breath condensate of normal controls" --- p.27 / Table2. Clinical and physiological details of the subjects --- p.29 / Figure 4. Level of eotaxin in exhaled breath condensate of asthma and control subjects --- p.30 / Figure 5 Level of MDC in exhaled breath condensate of asthma and control subjects --- p.31 / Table 3. Levels of eotaxin and MDC in exhaled breath condensate of asthma subjects on different dose of inhaled corticosteroids --- p.33 / Figure 6. Relationship between exhaled breath condensate level of MDC and total serum IgE level --- p.35

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A doença pulmonar obstrução crônica (DPOC) é caracterizada pela limitação de fluxo parcialmente reversível, classificada por níveis de obstrução pós-broncodilatador. Há várias evidências de que o FEV1 sozinho não é capaz de mostrar a broncodilatação de pacientes com DPOC, mesmo naqueles que apresentam melhora clínica. A técnica de oscilações forçadas (TOF) tem mostrado alta sensibilidade na detecção precoce de alterações mecânicas na DPOC, contudo o efeito broncodilatador na impedância respiratória de pacientes com DPOC ainda não está esclarecido. Objetiva avaliar a utilidade da TOF nos diferentes estágios de obstrução das vias aéreas; (2) avaliar a resposta da impedância respiratória ao salbutamol em indivíduos saudáveis ao exame espirométrico e pacientes com DPOC em diferentes graus de gravidade. Foram avaliados 25 indivíduos saudáveis sem história de tabagismo, 24 tabagistas e 151 pacientes com DPOC classificados em graus I, II, III e IV. Todos os sujeitos foram avaliados pela TOF seguida da espirometria, antes e após o uso do salbutamol spray. As curvas de resistência e reatância demonstraram alteração em todos os estágios de obstrução das vias aéreas após o uso do salbutamol. O grupo de risco apresentou alterações mecânicas semelhantes ao grupo leve (p=ns). Os parâmetros R0, Rm, Csr,din e Z4Hz apresentam desempenho diagnóstico adequado (AUC > 0,85) em todos os estágios de gravidade da doença. Todos os parâmetros de TOF e espirometria apresentaram diminuição após uso do salbutamol. Os indivíduos saudáveis apresentaram uma pequena diminuição comparada aos subgrupos de DPOC. A variação em termos absolutos da &#916;Z4Hz e das derivadas da resistência, &#916;R0, &#916;Rm, &#916;S, apresentaram variação significativa (p<0,0001, p<0,003; p<0,04; p<0,0002, respectivamente) com o aumento da obstrução brônquica. Nas derivadas da reatância o &#916;Xm aumentou com a gravidade da doença (p<0,0002). Por outro lado, a &#916;Crs,dyn não demonstrou diferença significativa com a gravidade da DPOC. Em termos percentuais os parâmetros da TOF apresentaram variação expressiva em &#916;Rm% (p<0,02), &#916;S% (p<0,02) e &#916;Xm% (p<0,004) com o aumento da obstrução nas vias aéreas. Por outro lado, &#916;R0%, &#916;Crs,dyn% e &#916;Z4Hz% não variaram entre os estágios da DPOC. A associação entre a broncodilatação nas vias aéreas e a impedância pulmonar foi fraca entre &#916;Xm vs &#916;FVC (r=0,32, p<0,0001) e &#916;Z4Hz% vs &#916;FEV1% vs &#916;FVC% (r=0.28, p<0,0005; r=0,29, p<0,0003, respectivamente). A TOF é útil na avaliação das alterações mecânicas nos diferentes níveis de obstrução das vias aéreas na DPOC. Demonstramos o benefício da medicação broncodilatadora, quantificando a melhora da ventilação através da TOF. A impedância respiratória diminui em todos os estágios da DPOC, o estágio leve melhorou tanto quanto o estágio muito grave. Isto sugere que a medida da impedância pulmonar não é dependente do volume como ocorre na espirometria e que a broncodilatação ocorre em todas as fases da progressão da DPOC. / Chronic obstructive pulmonary disease (COPD) is characterized by partially reversible flow limitation, classified by the post-bronchodilator level of airway obstruction. There is abundant evidence that FEV1 alone is not able to show bronchodilation in COPD patients, even in patients with clinical improvement. The forced oscillation technique (FOT) has shown high sensitivity for early detection of mechanical changes in COPD. However, the bronchodilator effect on respiratory impedance is still unclear. Objective to evaluate the utility of FOT in the diagnosis of different stages of airway obstruction, (2) to investigate the response to salbutamol in healthy, smoking and COPD patients in different degrees of severity. We evaluated 25 healthy subjects with no history of smoking, 24 smokers and 151 COPD patients classified into grades I, II, III and IV of severity. All subjects were assessed by the FOT followed by spirometry before and after the use of salbutamol spray. The resistance and reactance curves showed change at all stages of airway obstruction after the use of salbutamol. The smoking group showed similar mechanical changes to the mild group (p=ns). The parameters R0, Rm, Csr,din and Z4Hz presented adequate diagnostic accuracy (AUC>0.85) in all stages of disease severity. All FOT and spirometry parameters showed decreased after salbutamol use. Healthy individuals showed a small decrease compared with the subgroups of COPD. The variations of the impedance module (&#916;Z4Hz) and resistance parameters, (&#916;R0, &#916;Rm, &#916;s) were significant (p<0.0001, p<0.003, p<0.04 and p<0.0002, respectively) with increased bronchial obstruction. Mean reactance (&#916;Xm) increased with disease severity (p<0.0002). The &#916;Crs,dyn showed no significant change with the severity of COPD. In percentage terms, FOT parameters showed significant variation in &#916;Rm% (p<0.02), &#916;s (p<0.02) and &#916;Xm% (p<0.004) with increased airway obstruction. &#916;R0% &#916;Crs,dyn% and &#916;Z4Hz% did not vary between different stages of COPD. The association between bronchodilation in the airways and lung impedance was weak between &#916;Xm vs &#916;FVC (r=0.32, p<0.0001) and &#916;Z4Hz% vs &#916;FEV1% vs &#916;FVC% (r=0.28, p<0.0005, r=0.29, p<0.0003, respectively). The FOT is useful in the evaluation of the mechanical changes at different levels of airway obstruction in COPD. We demonstrate the benefit of a bronchodilator, quantifying the improvement of ventilation through the FOT. The respiratory impedance decreases in all stages of COPD. This suggests that the impedance changes are not dependent on lung volume as in spirometry and that bronchodilation occurs at all stages of the progression of COPD.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A doença pulmonar obstrução crônica (DPOC) é caracterizada pela limitação de fluxo parcialmente reversível, classificada por níveis de obstrução pós-broncodilatador. Há várias evidências de que o FEV1 sozinho não é capaz de mostrar a broncodilatação de pacientes com DPOC, mesmo naqueles que apresentam melhora clínica. A técnica de oscilações forçadas (TOF) tem mostrado alta sensibilidade na detecção precoce de alterações mecânicas na DPOC, contudo o efeito broncodilatador na impedância respiratória de pacientes com DPOC ainda não está esclarecido. Objetiva avaliar a utilidade da TOF nos diferentes estágios de obstrução das vias aéreas; (2) avaliar a resposta da impedância respiratória ao salbutamol em indivíduos saudáveis ao exame espirométrico e pacientes com DPOC em diferentes graus de gravidade. Foram avaliados 25 indivíduos saudáveis sem história de tabagismo, 24 tabagistas e 151 pacientes com DPOC classificados em graus I, II, III e IV. Todos os sujeitos foram avaliados pela TOF seguida da espirometria, antes e após o uso do salbutamol spray. As curvas de resistência e reatância demonstraram alteração em todos os estágios de obstrução das vias aéreas após o uso do salbutamol. O grupo de risco apresentou alterações mecânicas semelhantes ao grupo leve (p=ns). Os parâmetros R0, Rm, Csr,din e Z4Hz apresentam desempenho diagnóstico adequado (AUC > 0,85) em todos os estágios de gravidade da doença. Todos os parâmetros de TOF e espirometria apresentaram diminuição após uso do salbutamol. Os indivíduos saudáveis apresentaram uma pequena diminuição comparada aos subgrupos de DPOC. A variação em termos absolutos da &#916;Z4Hz e das derivadas da resistência, &#916;R0, &#916;Rm, &#916;S, apresentaram variação significativa (p<0,0001, p<0,003; p<0,04; p<0,0002, respectivamente) com o aumento da obstrução brônquica. Nas derivadas da reatância o &#916;Xm aumentou com a gravidade da doença (p<0,0002). Por outro lado, a &#916;Crs,dyn não demonstrou diferença significativa com a gravidade da DPOC. Em termos percentuais os parâmetros da TOF apresentaram variação expressiva em &#916;Rm% (p<0,02), &#916;S% (p<0,02) e &#916;Xm% (p<0,004) com o aumento da obstrução nas vias aéreas. Por outro lado, &#916;R0%, &#916;Crs,dyn% e &#916;Z4Hz% não variaram entre os estágios da DPOC. A associação entre a broncodilatação nas vias aéreas e a impedância pulmonar foi fraca entre &#916;Xm vs &#916;FVC (r=0,32, p<0,0001) e &#916;Z4Hz% vs &#916;FEV1% vs &#916;FVC% (r=0.28, p<0,0005; r=0,29, p<0,0003, respectivamente). A TOF é útil na avaliação das alterações mecânicas nos diferentes níveis de obstrução das vias aéreas na DPOC. Demonstramos o benefício da medicação broncodilatadora, quantificando a melhora da ventilação através da TOF. A impedância respiratória diminui em todos os estágios da DPOC, o estágio leve melhorou tanto quanto o estágio muito grave. Isto sugere que a medida da impedância pulmonar não é dependente do volume como ocorre na espirometria e que a broncodilatação ocorre em todas as fases da progressão da DPOC. / Chronic obstructive pulmonary disease (COPD) is characterized by partially reversible flow limitation, classified by the post-bronchodilator level of airway obstruction. There is abundant evidence that FEV1 alone is not able to show bronchodilation in COPD patients, even in patients with clinical improvement. The forced oscillation technique (FOT) has shown high sensitivity for early detection of mechanical changes in COPD. However, the bronchodilator effect on respiratory impedance is still unclear. Objective to evaluate the utility of FOT in the diagnosis of different stages of airway obstruction, (2) to investigate the response to salbutamol in healthy, smoking and COPD patients in different degrees of severity. We evaluated 25 healthy subjects with no history of smoking, 24 smokers and 151 COPD patients classified into grades I, II, III and IV of severity. All subjects were assessed by the FOT followed by spirometry before and after the use of salbutamol spray. The resistance and reactance curves showed change at all stages of airway obstruction after the use of salbutamol. The smoking group showed similar mechanical changes to the mild group (p=ns). The parameters R0, Rm, Csr,din and Z4Hz presented adequate diagnostic accuracy (AUC>0.85) in all stages of disease severity. All FOT and spirometry parameters showed decreased after salbutamol use. Healthy individuals showed a small decrease compared with the subgroups of COPD. The variations of the impedance module (&#916;Z4Hz) and resistance parameters, (&#916;R0, &#916;Rm, &#916;s) were significant (p<0.0001, p<0.003, p<0.04 and p<0.0002, respectively) with increased bronchial obstruction. Mean reactance (&#916;Xm) increased with disease severity (p<0.0002). The &#916;Crs,dyn showed no significant change with the severity of COPD. In percentage terms, FOT parameters showed significant variation in &#916;Rm% (p<0.02), &#916;s (p<0.02) and &#916;Xm% (p<0.004) with increased airway obstruction. &#916;R0% &#916;Crs,dyn% and &#916;Z4Hz% did not vary between different stages of COPD. The association between bronchodilation in the airways and lung impedance was weak between &#916;Xm vs &#916;FVC (r=0.32, p<0.0001) and &#916;Z4Hz% vs &#916;FEV1% vs &#916;FVC% (r=0.28, p<0.0005, r=0.29, p<0.0003, respectively). The FOT is useful in the evaluation of the mechanical changes at different levels of airway obstruction in COPD. We demonstrate the benefit of a bronchodilator, quantifying the improvement of ventilation through the FOT. The respiratory impedance decreases in all stages of COPD. This suggests that the impedance changes are not dependent on lung volume as in spirometry and that bronchodilation occurs at all stages of the progression of COPD.

L’hypertension artérielle pulmonaire (HTAP) est une maladie orpheline, progressive, incurable, caractérisée par un remodelage intense de la paroi des artérioles pulmonaires résistives conduisant à l’augmentation des résistances vasculaires pulmonaires. Des anomalies génétiques associées à des mutations du gène codant pour le récepteur de type 2 de morphogenèse osseuse (BMPR2) ont été mises en évidence chez près de 70% des formes héréditaires d’HTAP. Dans le présent travail et ce à l’aide de cultures cellulaires primaires réalisées à partir de biopsies pulmonaires collectées chez des patients souffrant d’HTAP idiopathique et des sujets contrôles, nous nous sommes intéressés aux mécanismes responsables de l’hyperplasie des cellules musculaires lisses présente dans l’HTAP, plus particulièrement à l’implication de la voie angiopoîétine (Ang)/Tie2 dans la communication intercellulaire existant entre cellules endothéliales (CE) et cellules musculaires lisses artérielles pulmonaires (CML-AP), ainsi qu’à la voie des BMP/BMPR2 chez des patients HTAP porteurs de mutations BMPR2. <p>Contrairement aux expressions inchangées des Ang1 et Ang2, l’expression du récepteur endothélial Tie2 est augmentée tant au niveau pulmonaire que cellulaire dans l’HTAP. Les CML-AP isolées à partir de patients souffrant d’HTAP présentent in vitro un phénotype prolifératif accru et ce d’autant plus en présence de surnageants de culture collectés à partir de CE pulmonaires (CE-P). Cet effet prolifératif est d’autant plus intense que les CE-P proviennent de patients HTAP et qu’elles ont été préalablement stimulées par l’Ang1. La fluoxétine, un inhibiteur du transporteur de la sérotonine (5-HT) inhibe ces effets prolifératifs, alors que le bosentan, un antagoniste non sélectif des récepteurs A et B à l’endothéline (ET) n’a pas d’effet. De plus, l’activation du récepteur Tie2 induit la synthèse et la libération de 5-HT et d’ET1 par les CE-P isolées à partir de patients souffrant d’HTAP. La voie angiopoïétine/Tie2 joue donc un rôle central dans le dialogue croisé existant entre l’endothélium et le muscle lisse vasculaire pulmonaire et son activation contribue à la prolifération accrue des CML-AP responsable du remodelage vasculaire pulmonaire chez les patients souffrant d’HTAP idiopathique. <p>L’expression du BMPR2 est diminuée tant au niveau pulmonaire, que cellulaire chez les patients souffrant d’HTAP et porteurs de mutations BMPR2. Dans ces CML-AP mutées, le BMP4 ne freine pas la prolifération et n’induit pas l’apoptose, normalement induites dans les cellules isolées à partir de contrôles et de patients souffrant d’HTAP non porteurs de mutations BMPR2. Ces particularités fonctionnelles sont associées à une activation de la voie p38MAPK et un défaut d’activation de la voie Smad. Parmi les 9 mutations BMPR2 étudiées, seule la mutation par délétion de 22 paires de bases dans le domaine transmembranaire ne présente pas ces particularités fonctionnelles. Des anomalies génétiques, telles les mutations BMPR2 concourent donc à une perte du contrôle de la prolifération cellulaire associé à l’activation de la voie p38MAPK, présentant les mutations BMPR2 étudiées, comme des facteurs de risque supplémentaires au développement de l’hyperplasie médiale dans l’HTAP. <p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

MCur / Department of Advanced Nursing Science

Indiana University-Purdue University Indianapolis (IUPUI) / The patients with Cystic fibrosis (CF), an inherent genetic disorder, suffer from chronic bacterial infection in the lung. In CF, modification of epithelial cells leads to alteration of the lung environment, such as inhibition of ciliary bacterial clearance and accumulation of thickened mucus in the airways. Exploiting these conditions, opportunistic pathogens like Pseudomonas aeruginosa cause lifelong persistent infection in the CF lung by forming into antibiotic-resistant aggregated communities called biofilms. Airway infections as well as inflammation are the two major presentations of CF lung disease. P. aeruginosa strains isolated from CF lungs often contain mutations in the mucA gene, and this mutation results in higher level expression of bacterial polysaccharides and toxic lipoproteins. In a previous work, we have found a putative lipoprotein gene (PA4326) which is overexpressed in antibiotic-induced biofilm formed on cultured CF-derived airway cells. In the current work, we speculated that this particular putative lipoprotein affects cellular cytotoxicity and immune-stimulation in the epithelial cells. We found that mutation of this gene (ΔPA4326) results in reduced airway cell killing without affecting other common virulence factors.Moreover, we observed that this gene was able to stimulate secretion of the proinflammatory cytokine IL-8 from host cells. Interestingly, we also found that ΔPA4326 mutant strains produced less pyocyanin exotoxin compared to the wild type. Furthermore, our results suggest that PA4326 regulates expression of the pyocyanin biosynthesis gene phzM, leading to the reduced pyocyanin phenotype. Overall, these findings implicate PA4326 as a virulence factor in Pseudomonas aeruginosa. In the future, understating the molecular interplay between the epithelial cells and putative lipoproteins like PA4326 may lead to development of novel anti-inflammatory therapies that would lessen the suffering of CF patients.

In developing countries, acute respiratory infections (ARIs) are responsible for two million deaths per year. Most victims are children who are less than 5 years old. Pneumonia kills 5000 children per day. The statistics for cardiovascular diseases (CVDs) are even more alarming. According to a 2009 report from the World Health Organization (WHO), CVDs kill 17 million people per year. In many resource-poor parts of the world such as India and China, many people are unable to access cardiologists, pulmonologists, and other specialists. Hence, low skilled health professionals are responsible for screening people for ARIs and CVDs in these areas. For example, in the rural areas of the Philippines, there is only one doctor for every 10,000 people. By contrast, the United States has one doctor for every 500 Americans. Due to advances in technology, it is now possible to use a smartphone for audio recording, signal processing, and machine learning. In my thesis, I have developed an Android application named Smart Auscultation. Auscultation is a process in which physicians listen to heart and lung sounds to diagnose disorders. Cardiologists spend years mastering this skill. The Smart Auscultation application is capable of recording and classifying heart sounds, and can be used by public or clinical health workers. This application can detect abnormal heart sounds with up to 92-98% accuracy. In addition, the application can record, but not yet classify, lung sounds. This application will be able to help save thousands of lives by allowing anyone to identify abnormal heart and lung sounds.

Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished