In vitro cytotoxicity of metal ions and roadside dust collected in Hong Kong.

Lau Wing-Ngar Vivian.Thesis (M.Phil.)--Chinese University of Hong Kong, 2002.Includes bibliographical references (leaves 135-144).Abstracts in English and Chinese.Acknowledgements --- p.iAbstract --- p.iiAbbreviations --- p.viList of figures --- p.viiiList of tables --- p.xiContents --- p.xiiiChapter 1 --- Introduction --- p.1Chapter 1.1 --- General introduction --- p.1Chapter 1.2 --- Roadside air pollution worldwide and in Hong Kong --- p.2Chapter 1.2.1 --- Air quality in Hong Kong --- p.3Chapter 1.3 --- Characteristics of particulate matter --- p.9Chapter 1.4 --- Composition and sources of particulate matter --- p.11Chapter 1.5 --- Toxic effects of particulate matter --- p.12Chapter 1.5.1 --- Lung injury --- p.12Chapter 1.5.2 --- Cardiovascular injury --- p.15Chapter 1.5.3 --- Mutagenesis and carcinogenesis --- p.16Chapter 1.6 --- Aims of my study --- p.16Chapter 2 --- Toxic Effects of Heavy Metals Ions on Selected Cultured Cell-lines --- p.18Chapter 2.1 --- Introduction --- p.18Chapter 2.1.1 --- Metals --- p.18Chapter 2.1.1.1 --- Cadmium --- p.22Chapter 2.1.1.2 --- Chromium --- p.23Chapter 2.1.1.3 --- Lead --- p.25Chapter 2.1.1.4 --- Zinc --- p.26Chapter 2.1.2 --- Metallothioneins --- p.28Chapter 2.1.3 --- p53 --- p.31Chapter 2.1.4 --- Tumor Necrosis Factor-alpha (TNF-α) --- p.32Chapter 2.1.5 --- Aims of this chapter --- p.32Chapter 2.2 --- Materials and methods --- p.35Chapter 2.2.1 --- Reagents --- p.35Chapter 2.2.2 --- Cultured Cell lines --- p.35Chapter 2.2.2.1 --- PU5-18 --- p.36Chapter 2.2.2.2 --- LL24 --- p.36Chapter 2.2.2.3 --- HBE4-E6/E7 --- p.37Chapter 2.2.3 --- Cytotoxicity assays --- p.37Chapter 2.2.4 --- ELISA assays --- p.40Chapter 2.2.4.1 --- ELISA assay ofp53 levels --- p.41Chapter 2.2.4.2 --- ELISA assay of TNF-α levels --- p.43Chapter 2.2.5 --- MT gene expression studies by Luciferase assay --- p.44Chapter 2.2.5.1 --- PCR amplification --- p.44Chapter 2.2.5.2 --- 5´ة End modification of PCR amplified DNA --- p.44Chapter 2.2.5.3 --- Ligation of DNA fragment to linearized vector --- p.46Chapter 2.2.5.4 --- E. coli. transformation by heat shock --- p.46Chapter 2.2.5.5 --- PCR sequencing --- p.47Chapter 2.2.5.6 --- Transfection of plasmid into HBE4-E6/E7 cells --- p.49Chapter 2.2.5.7 --- Data analysis --- p.50Chapter 2.3 --- Results and discussion --- p.51Chapter 2.3.1 --- Cytotoxicity assays --- p.51Chapter 2.3.2 --- Combination effects of metals on cytotoxicity --- p.61Chapter 2.3.3 --- p53 --- p.65Chapter 2.3.4 --- TNF-α --- p.68Chapter 2.3.5 --- MT gene expression studies by Luciferase assay --- p.69Chapter 2.4 --- Conclusion --- p.74Chapter 3 --- Effects of Polycyclic Aromatic Hydrocarbons (PAHs) on Cultured Cell-lines --- p.75Chapter 3.1 --- Introduction --- p.75Chapter 3.2 --- Materials and methods --- p.79Chapter 3.2.1 --- Reagents --- p.79Chapter 3.2.2 --- Cell culture --- p.79Chapter 3.2.3 --- AlamarBlue assay --- p.80Chapter 3.2.4 --- EROD assay --- p.80Chapter 3.3 --- Results and discussion --- p.84Chapter 3.4 --- Conclusion --- p.88Chapter 4 --- Chemical and Biological Assays on Roadside Dust --- p.89Chapter 4.1 --- Introduction --- p.89Chapter 4.1.1 --- Composition of particulate matter in Hong Kong --- p.89Chapter 4.1.2 --- Metal contents of particulate matter in Hong Kong --- p.91Chapter 4.1.3 --- Possible adverse health impacts of particulate matter --- p.94Chapter 4.1.3.1 --- In vitro studies using different cell models --- p.94Chapter 4.1.3.2 --- In vivo studies using rodents --- p.97Chapter 4.1.3.3 --- Epidemiological studies --- p.98Chapter 4.1.4 --- Aims of this chapter --- p.100Chapter 4.2 --- Materials and methods --- p.101Chapter 4.2.1 --- Sampling of roadside dust --- p.101Chapter 4.2.2 --- Chemical analysis of roadside dust --- p.104Chapter 4.2.2.1 --- Reagents --- p.104Chapter 4.2.2.2 --- Total metal contents --- p.105Chapter 4.2.2.3 --- Extractable metal contents --- p.105Chapter 4.2.3 --- Biological assays --- p.105Chapter 4.2.3.1 --- Cell models --- p.106Chapter 4.2.3.2 --- Pretreatment of roadside dust --- p.106Chapter 4.2.3.3 --- AlamarBlue assay --- p.106Chapter 4.2.3.4 --- ELISA assays --- p.108Chapter 4.2.3.5 --- Luciferase assay --- p.108Chapter 4.3 --- Results and discussion --- p.110Chapter 4.3.1 --- Total metal contents --- p.110Chapter 4.3.2 --- Extractable metal contents --- p.113Chapter 4.3.3 --- AlamarBlue assay --- p.116Chapter 4.3.4 --- p53 --- p.122Chapter 4.3.5 --- TNF-α --- p.122Chapter 4.3.6 --- Luciferase assay --- p.126Chapter 4.4 --- Conclusion --- p.129Chapter 5 --- General discussion and conclusion --- p.130Chapter 6 --- References --- p.13