Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.

M. Tech. / The persistent presence of neutrophils is associated with a wide range of inflammatory diseases. Resolution of inflammation in these diseases is also associated with the ingestion of apoptotic neutrophils by macrophages. Inflammation and apoptosis of inflammatory cells are common known features observed in the lung following exposure to crystalline silica. What is not known is how well these apoptotic cells are cleared by macrophages in the presence of crystalline silica? To investigate the latter, we incubated the U937 macrophages and neutrophils with crystalline silica and found that it could increase their apoptosis and necrosis especially those of the U937 cells. We then examined the ability of crystalline silica to induce the production of cytokines (TNF-α, IFN-γ and IL-1β) as well as NO by these cells. We found that these particles could increase the production of TNF-α, IL-1β and NO but not IFN-γ in a time-and concentration-dependent manner. We also assessed the ability of crystalline silica to alter the levels of GSH in neutrophils and U937 macrophages. We found that it could drastically decrease the levels of this antioxidant in U937 macrophages with no additional effect in neutrophils as these latter cells would have low levels of GSH prior to their incubation with crystalline silica. Finally, we examined the effect of crystalline silica on the ability of U937 macrophages to phagocytose apoptotic neutrophils. We found that while untreated U937 macrophages were able to phagocytose apoptotic neutrophils, the presence of crystalline silica reduced this ability by 15%. Taken together, our results suggest that exposure to crystalline silica impairs the clearance of apoptotic neutrophils by decreasing their phagocytosis by macrophages and thus prevents the resolution of inflammation.

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

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2005. / Härtill 4 uppsatser.

Ng Sin Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 120-135). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abbreviations --- p.iii / Abstract --- p.vi / 撮要 --- p.ix / Publications --- p.xi / Table of contents --- p.xiii / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Allergic Diseases and Allergic Inflammation --- p.1 / Chapter 1.1.1 --- Prevalence of Allergic Diseases --- p.1 / Chapter 1.1.2 --- Common Allergic Diseases: Allergic Asthma --- p.1 / Chapter 1.1.3 --- Common Allergic Diseases: Atopic Dermatitis --- p.2 / Chapter 1.1.4 --- Allergic Inflammation --- p.3 / Chapter 1.2 --- The Inflammatory Leukocytes: Mast Cells and Eosinophils --- p.6 / Chapter 1.2.1 --- Characteristics of Mast Cells --- p.6 / Chapter 1.2.2 --- Mast Cells Distribution --- p.8 / Chapter 1.2.3 --- Mast Cells Subtypes --- p.8 / Chapter 1.2.4 --- HMC-1 Cells --- p.9 / Chapter 1.2.5 --- Characteristics of Eosinophils --- p.12 / Chapter 1.3 --- Adhesion Molecules in Allergic Diseases --- p.15 / Chapter 1.3.1 --- Adhesion Molecules and Leukocyte Migration --- p.15 / Chapter 1.3.2 --- Selectin --- p.17 / Chapter 1.3.3 --- Intermolecular Adhesion Molecules --- p.17 / Chapter 1.3.4 --- Integrin --- p.18 / Chapter 1.4 --- Cytokines and Chemokines in Allergic Diseases --- p.18 / Chapter 1.4.1 --- IL-6 --- p.20 / Chapter 1.4.2 --- CXCL1 --- p.21 / Chapter 1.4.3 --- CXCL8 --- p.21 / Chapter 1.4.3 --- CCL2 --- p.22 / Chapter 1.5 --- Intercellular Signal Transduction Pathways in Inflammation --- p.24 / Chapter 1.5.1 --- RAS-RAF-mitogen-activated Protein Kinases --- p.24 / Chapter 1.5.2 --- Janus Kinase/ Signal Transducers and Activators of Transcriptions Pathway --- p.27 / Chapter 1.5.3 --- Nuclear Factor-KB Pathway --- p.29 / Chapter 1.5.4 --- Phosphoinositide 3-Kinase Pathway --- p.31 / Chapter 1.6 --- Aims and Scope of the Study --- p.33 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials --- p.35 / Chapter 2.1.1 --- HMC-1 Cell Line --- p.35 / Chapter 2.1.2 --- Human Buffer Coat --- p.35 / Chapter 2.1.3 --- Human Mast Cell Chymase and TLR ligands --- p.35 / Chapter 2.1.4 --- Media and Reagents for Cell Culture --- p.36 / Chapter 2.1.5 --- Reagents and Buffers for Purification of Human Eosinophils --- p.37 / Chapter 2.1.6 --- Reagents and Buffers for Flow Cytmetry --- p.38 / Chapter 2.1.7 --- Reagents and Buffers for Total RNA Extraction --- p.41 / Chapter 2.1.8 --- Reagents and Buffers for Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.42 / Chapter 2.1.9 --- Reagents and Buffers for Agarose Gel Electrophoresis --- p.45 / Chapter 2.1.10 --- Reagents and Buffers for Sodium Dodecyl Sulfate -polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.46 / Chapter 2.1.11 --- Reagents and Buffers for Western Blot Analysis --- p.48 / Chapter 2.1.12 --- Chemotactic Migration --- p.51 / Chapter 2.1.13 --- Signaling Transduction Inhibitors and Protein Synthesis Inhibitors --- p.51 / Chapter 2.2 --- Methods --- p.52 / Chapter 2.2.1 --- HMC-1 Cell Cultures --- p.52 / Chapter 2.2.2 --- Purification of Buffy Coat Eosinophils by MACS and Eosinophil Culture --- p.52 / Chapter 2.2.3 --- Total Cellular RNA Extraction --- p.53 / Chapter 2.2.4 --- RT-PCR --- p.54 / Chapter 2.2.5 --- Agarose Gel Electrophoresis --- p.55 / Chapter 2.2.6 --- Flow Cytometry Analysis --- p.55 / Chapter 2.2.7 --- Protein Array Analysis of Cytokine Release --- p.57 / Chapter 2.2.8 --- Quantitative Analysis ofCXCLl --- p.58 / Chapter 2.2.9 --- Total Protein Extraction --- p.58 / Chapter 2.2.10 --- SDS-PAGE --- p.58 / Chapter 2.2.11 --- Western Blot Analysis --- p.59 / Chapter 2.2.12 --- Chemotactic Migration Analysis --- p.60 / Chapter 2.2.13 --- Statistical Analysis --- p.60 / Chapter Chapter 3 --- Effects of Mast Cell Derived Chymase on Human Eosinophils and the Signaling Mechanisms: Implication in Allergic Inflammation / Chapter 3.1 --- Introduction --- p.61 / Chapter 3.2 --- Results --- p.65 / Chapter 3.2.1 --- Effects of Chymase on Eosinophil Survival --- p.65 / Chapter 3.2.2 --- Effects of Chymase on the Adhesion Molecule Expression of Eosinophils --- p.68 / Chapter 3.2.3 --- Effects of Chymase on the Chemokinetic Properties on Eosinophils --- p.71 / Chapter 3.2.4 --- Effects of Chymase on the Release of Chemokines and IL-6 from Eosinophils --- p.73 / Chapter 3.2.5 --- Signal Transduction Mechanism Involved in Regulating Chymase-induced Effects on Eosinophils --- p.78 / Chapter 3.3 --- Discussion --- p.71 / Chapter Chapter 4 --- TLR-mediated Effects and Signal Transduction Mechanism of HMC-1 Cells / Chapter 4.1 --- Introduction --- p.92 / Chapter 4.2 --- Results --- p.97 / Chapter 4.2.1 --- Expression of Adhesion Molecules on HMC-1 Cells --- p.95 / Chapter 4.2.2 --- TLR Expression Profile on HMC-1 Cells --- p.97 / Chapter 4.2.3 --- Effects of TLR ligands on HMC-1 Cell Adhesion Molecule Expressions --- p.99 / Chapter 4.2.4 --- TLR7-induced Phosphorylation of ERK and Effects of PD98059 on TLR7-induced ERK Phosphorylation --- p.104 / Chapter 4.2.5 --- Effect of TLR7 Ligand on HMC-1 Cells Cytokine Release --- p.108 / Chapter 4.3 --- Discussion --- p.110 / Chapter Chapter 5 --- Conclusions and Future Perspectives / Chapter 5.1 --- Conclusions --- p.115 / Chapter 5.2 --- Future Perspectives --- p.117 / References --- p.120 / Appendix --- p.136

Abstract not available

Includes bibliographical references (leaves 182-240). Explores alternate strategies that may alter inflammatory cytokine production, particularly tumour necrosis factor đ [tumor necrosis factor-alpha], and therefore provide a possible treatment for rheumatoid arthritis.

Includes bibliographical references. / xvii, 228 leaves : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Methods were developed to observe and quantify neutrophil transendothelial migration, both together with or independently of neutrophil-endothelial adhesion. Two mechanicisms of neutrophil transmigration were identified, and the role of one of the major mediators of this process, IL-8, was described. / Thesis (Ph.D.)--University of Adelaide, Dept. of Medicine 1994

Inflammation is the normal biological response to tissue injury, and is characterised by the interactive activation of multiple mediators and cell types. One response to tissue injury is the production of pain, not only by direct trauma to sensory fibres, but also through the release of mediators from sensory nerve terminals. One such mediator is kinin which is a vasoactive peptide considered to play a primary role in inflammation by causing constriction of venules, dilation of arterioles, increasing permeability of the capillary membrane, and interacting with sensory nerve terminal transmitters to evoke pain. The kinin forming enzymes (kallikreins) reach inflammation sites either on the surface of migrating neutrophils or by transudation from plasma. The kininogen molecule which contains the kinin moiety, has been localised on the external surface of the neutrophil, and provides the substrate from which kinins can be cleaved through enzymatic action. The cellular actions of kinins are mediated through B2 receptors, which are also located on the external surface of the neutrophils. In addition, the induced effects of kinins are regulated by B1 receptors. The formation of nitric oxide (NO) from arginine released from the kinin C terminus, and receptor membrane signal transduction by nitric oxide following kinin receptor activation is discussed. A molecular response to cell injury is the formation of chemotactic mediators that attract neutrophils to sites of inflammation. The question whether neutrophils contribute to circulating levels of kinins was examined in infections and inflammatory disorders. This novel hypothesis was tested using circulating neutrophils harvested from patients with tuberculosis meningitis and pneumonia. These neutrophils showed a distinct loss of only the kinin moiety from the kininogen molecule located on the external surface. The confocal images of fixed, permeabilised neutrophils provided multi-dimensional constructs, and the intensity of fluorescence reflected the relative amounts of the molecule present in both neutrophils harvested from healthy volunteers as well as patient blood. The immunocytochemical labelling experiments using colloidal gold as markers, confirmed, at the ultrastructural level, the presence or disappearance of the kinin moiety from the kininogen molecule on the neutrophil surface. The cell component of synovial fluid in rheumatoid athritis (RA) consists mainly of neutrophils. This study demonstrates the absence of the kinin moiety from circulating and synovial fluid neutrophils from patients with RA, as well as an increased signal from immunolabelled B2 receptors in synovial fluid neutrophils. These findings support the hypothesis that in RA, kinins are released during the inflammatory response in the joints, and suggests that there is an upregulation of the B2 receptor at the site of inflammation. Neutrophils chemotactically drawn to the site of inflammation become activated to release kinin from the kininogen molecule, and thereafter re-enter the circulation where they were harvested systemically. B2 receptors may be upregulated following activation by kinins or by other mediators present in the inflammatory milieu. Interleukin-1 has been shown to upregulate kinin receptors on human synovial cells. Anti-peptide antibodies to the loops of cloned B1 and B2 receptors have provided powerful probes for the cellular identification of the two kinin receptor families. Mapping of the B2 receptors showed upregulation on the neutrophils gathered from inflamed joints. However, no activation of the Br receptors was observed in normal blood neutrophils as well as those obtained from the different disease states. / Thesis (M.Med.)-University of Natal, 1996.

Typescript (Photocopy) / Bibliography: leaves 202-281 / x, 281 leaves, 2 leaves of plates : ill ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--Dept. of Medicine, University of Adelaide, 1990