by Henry, Kam Yin Cheung. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 148-160). / ZEBRAFISH (BRACHYDANIO RERIO) / ACKNOWLEDGEMENTS / LIST OF CONTENTS / ABSTRACT / ABBREVIATION / Chapter CHAPTER ONE --- INTRODUCTION / Chapter 1.1 --- Transgenic fish --- p.1 / Chapter 1.2 --- Zebrafish --- p.4 / Chapter 1.3 --- The grass carp GH gene and protein / Chapter 1.3.1 --- The genomic sequence --- p.5 / Chapter 1.3.2 --- The cDNA sequence --- p.7 / Chapter 1.3.3 --- The grass carp GH protein --- p.7 / Chapter 1.4 --- Functional aspects of promoter regions / Chapter 1.4.1 --- PEPCK --- p.9 / Chapter 1.4.2 --- RSV-LTR --- p.10 / Chapter 1.4.3 --- hMT-IIA --- p.10 / Chapter 1.4.4 --- MMTV-LTR --- p.11 / Chapter 1.5 --- Eukaryotic gene expression in cultured cells / Chapter 1.5.1 --- COS-7 and HepG2 cells --- p.11 / Chapter 1.5.2 --- Transfection system --- p.12 / Chapter 1.5.3 --- Fate of DNA after transfection --- p.13 / Chapter 1.6 --- Electroporation and microinjection as tools for gene transfer / Chapter 1.6.1 --- Electroporation: Theory and operation --- p.13 / Chapter 1.6.2 --- Microinjection: Design of microinjector --- p.16 / Chapter 1.6.3 --- Fate of DNA after gene transfer in embryos / Transient expression --- p.16 / Stable transformation --- p.17 / Inheredity of transgene --- p.17 / Chapter 1.7 --- The aims of the present study --- p.18 / Chapter CHAPTER TWO --- MATERIALS AND METHODS / Chapter 2.1 --- General techniques / Chapter 2.1.1 --- Electrophoresis of DNA / Chapter 2.1.1.1 --- Agarose gel electrophoresis --- p.19 / Chapter 2.1.1.2 --- PAGE --- p.20 / Chapter 2.1.2 --- Purification of DNA --- p.21 / Chapter 2.1.3 --- Recovery of DNA fragments / Chapter 2.1.3.1 --- Electroelution --- p.22 / Chapter 2.1.3.2 --- Geneclean kit --- p.23 / Chapter 2.1.4 --- Standard recombinant DNA techniques / Chapter 2.1.4.1 --- Dephosphorylation --- p.24 / Chapter 2.1.4.2 --- Kinasing --- p.24 / Chapter 2.1.4.3 --- Ligation --- p.24 / Chapter 2.1.4.4 --- Filling in reaction --- p.25 / Chapter 2.1.4.5 --- Transformation --- p.25 / Chapter 2.1.5 --- Minipreparation of plasmids --- p.26 / Chapter 2.1.6 --- Large preparation of plasmids / Chapter 2.1.6.1 --- Qiagene kit --- p.27 / Chapter 2.1.6.2 --- CsCl density gradient centrifugation --- p.27 / Chapter 2.1.7 --- DNA sequencing --- p.29 / Chapter 2.1.8 --- "Extraction of DNA from embryos, fry and fish" / Method 1 --- p.32 / Method 2 --- p.32 / Chapter 2.1.9 --- Probe labelling / Chapter 2.1.9.1 --- End-labelling --- p.33 / Chapter 2.1.9.2 --- Random priming --- p.33 / Chapter 2.1.10 --- CAT assay --- p.33 / Chapter 2.1.11 --- Polymerase chain reaction(PCR) --- p.35 / Chapter 2.1.12 --- Radioimmunassay(RIA) of FGH --- p.36 / Chapter 2.1.13 --- Dot blotting --- p.38 / Chapter 2.1.14 --- Southern blotting --- p.39 / Chapter 2.2 --- "Linkers, primers and probes" / Chapter 2.2.1 --- Primers --- p.41 / Chapter 2.2.2 --- Linkers --- p.45 / Chapter 2.2.3 --- Probes --- p.47 / Chapter 2.3 --- Construction of chimeric growth hormone genes / Chapter 2.3.1 --- Sources of plasmids --- p.50 / Chapter 2.3.2 --- General principles --- p.50 / Chapter 2.3.3 --- PEPCKgcGHcDNA --- p.51 / Chapter 2.3.4 --- RSVgcGHcDNA --- p.54 / Chapter 2.3.5 --- hMTgcGHcDNAcDNA --- p.56 / Chapter 2.3.6 --- MMTVgcGHcDNA --- p.58 / Chapter 2.3.7 --- "PEPCKgcGH, RSVgcGH and hMTgcGH" --- p.60 / Chapter 2.4 --- Expression of chimeric genes in cultured cells / Chapter 2.4.1 --- Culturing of COS-7 and HepG2 cells --- p.66 / Chapter 2.4.2 --- Expression of chimeric genes in COS-7 and HepG2 cells --- p.67 / Chapter 2.5 --- Zebrafish / Chapter 2.5.1 --- "Culturing, Spawning and hatching" --- p.67 / Chapter 2.6 --- Electroporation and microinjection for gene transfer / Chapter 2.6.1 --- Electroporation / Chapter 2.6.1.1 --- Tuning up electroporation --- p.69 / Chapter 2.6.1.2 --- Evidence of gene transfer by electroporation / Chapter 2.6.1.2.1 --- CAT assay --- p.71 / Chapter 2.6.1.2.2 --- Dot blot --- p.71 / Chapter 2.6.1.2.3 --- PCR and Southern blotting of PCR products --- p.72 / Chapter 2.6.1.2.4 --- Southern blotting of fish total DNA --- p.73 / Chapter 2.6.2 --- Microinjection / Chapter 2.6.2.1 --- Handling of microinjection --- p.74 / Chapter 2.6.2.2 --- Evidence of gene transfer by microinjection / Chapter 2.6.2.2.1 --- CAT assay --- p.75 / Chapter 2.6.2.2.2 --- PCR and Southern blotting of PCR products --- p.75 / Chapter 2.7 --- Phenotypic alteration of fish generated from electroporated eggs / Chapter 2.7.1 --- Electroporation and handling of fish generated from electroporation --- p.75 / Chapter 2.7.2 --- Measurement of phenotypic change in fish generated from electroporation --- p.77 / Chapter 2.8 --- Detection of transgene and expression of exogenous DNA / Chapter 2.8.1 --- Transgene detection --- p.78 / Chapter 2.8.2 --- Expression of exogenous DNA --- p.79 / Chapter CHAPTER THREE --- RESULTS / Chapter 3.1 --- Construction of Chimeric growth hormone genes / Chapter 3.1.1 --- Confirmation of integrity of chimeric genes / PEPCKgcGHcDNA --- p.80 / RSVgcGHcDNA --- p.81 / hMTgcGHcDNA --- p.81 / MMTVgcGHcDNA --- p.81 / "PEPCKgcGH, RSVgcGH and hMTgcGH" --- p.82 / Chapter 3.1.2 --- Yield of chimeric genes from CsCl density gradient centrifugation --- p.82 / Chapter 3.2 --- Chimeric gene expression in COS-7 and HepG2 cells / Chapter 3.2.1 --- Expression of chimeric genes in COS-7 cells --- p.89 / Chapter 3.2.2 --- Expression of chimeric genes in HepG2 cells --- p.93 / Chapter 3.3 --- Transfer of chimeric genes into embryos / Chapter 3.3.1 --- Electroporation / Chapter 3.3.1.1 --- Monitoring of electroporation --- p.94 / Chapter 3.3.1.2 --- Evidence for gene transfer / Chapter 3.3.1.2.1 --- CAT assay --- p.98 / Chapter 3.3.1.2.2 --- Dot blotting --- p.98 / Chapter 3.3.1.2.3 --- PCR and Southern blotting of PCR product --- p.101 / Chapter 3.3.1.2.4 --- Southern blotting of DNA from fish generated from electroporation --- p.106 / Chapter 3.3.2 --- Microinjection / Chapter 3.3.2.1 --- CAT assay --- p.109 / Chapter 3.3.2.2 --- PCR --- p.109 / Chapter 3.4 --- Phenotypic alterations of fish / The first experiment --- p.112 / The second experiment --- p.113 / The third experiment --- p.113 / The fourth experiment --- p.122 / Chapter 3.5 --- Detection of transgene and expression of exogenous DNA / Chapter 3.5.1 --- Transgene --- p.128 / Chapter 3.5.2 --- Possible expression of exogenous DNA --- p.129 / Chapter CHAPTER FOUR --- DISCUSSION / Chapter 4.1 --- Chimeric growth hormone genes --- p.132 / Chapter 4.2 --- Expression of chimeric growth hormone genes in COS-7 and HepG2 cells --- p.134 / Chapter 4.3 --- Transfer of exogenous DNA into embyros --- p.136 / Chapter 4.4 --- Phenotypic alteration of fish developed from electroporated eggs --- p.139 / Chapter 4.5 --- The possible integration and expression of exogenous DNA --- p.143 / Chapter 4.6 --- Conclusions --- p.145 / Chapter 4.7 --- Suggestions for further studies --- p.146 / REFERENCES --- p.148 / Chapter APPENDIX I --- Restriction maps / PEPCKgcGH / PEPCKgcGHcDNA / RSVgcGH / RSVgcGHcDNA / hMTgcGH / hMTgcGHcDNA / MMTVgcGHcDNA / pBH1.2 / pMSG-CAT / pUC19 / hMT-IIA / PBC12BI / RSVCAT / pUC101 / pSEl/S2 / PUCSE2/S1 / pUCS2

by Wai Ching Wa, Gina. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1990. / Bibliography: leaves 303-314. / Chapter 1. --- INTRODUCTION --- p.1 / Chapter 2. --- LITERATURE REVIEW ON CERVIX UTERI / Chapter 2.1 --- HISTOLOGY OF CERVIX UTERI --- p.4 / Chapter 2.2 --- CYTOLOGY OF CERVIX UTERI --- p.5 / Chapter 2.3 --- CERVICAL EPITHELIAL LESIONS / Chapter 2.3.1 --- Squamous Metaplasia --- p.7 / Chapter 2.3.2 --- Cervical Intraepithelial Neoplasia --- p.9 / Chapter 2.3.3 --- Viral Infections --- p.17 / Chapter 2.4 --- DIAGNOSTIC APPROACH TO CERVICAL LESIONS --- p.20 / Chapter 2.5 --- DIAGNOSTIC VARIABILITY OF CERVICAL LESIONS --- p.21 / Chapter 2.6 --- IMPORTANCE OF CERVICAL CARCINOMA IN HONG KONG --- p.21 / Chapter 3. --- LITERATURE REVIEW ON NASOPHARYNX / Chapter 3.1 --- HISTOLOGY OF NASOPHARYNX --- p.22 / Chapter 3.2 --- CYTOLOGY OF NASOPHARYNX --- p.24 / Chapter 3.3 --- NASOPHARYNGEAL EPITHELIAL LESIONS / Chapter 3.3.1 --- 'Squamous Metaplasia' --- p.25 / Chapter 3.3.2 --- Nasopharyngeal Intraepithelial Neoplasia --- p.26 / Chapter 3.3.3 --- Viral Infections --- p.27 / Chapter 3.4 --- DIAGNOSTIC APPROACH TO NASOPHARYNGEAL LESIONS --- p.28 / Chapter 3.5 --- IMPORTANCE OF NASOPHARYNGEAL CARCINOMA IN HONG KONG --- p.29 / Chapter 4. --- LITERATURE REVIEW ON MORPHOMETRY / Chapter 4.1 --- QUANTITATIVE ASSESSMENT OF CELL FEATURES --- p.30 / Chapter 4.2 --- TERMINOLOGY --- p.31 / Chapter 4.3 --- APPROACHES TO SAMPLING --- p.32 / Chapter 4.4 --- SOURCES OF VARIATION --- p.32 / Chapter 4.5 --- METHODOLOGY FOR MORPHOMETRY --- p.33 / Chapter 4.6 --- FEATURES FOR MORPHOMETRY IN INTRAEPITHELIAL NEOPLASIA --- p.35 / Chapter 4.7 --- PREVIOUS MORPHOMETRIC STUDIES ON INTRAEPITHELIAL NEOPLASIA --- p.36 / Chapter 5. --- MATERIALS AND METHODS / Chapter 5.1 --- MATERIALS / Chapter 5.1.1 --- Cervix Uteri --- p.41 / Chapter 5.1.2 --- Nasopharynx --- p.41 / Chapter 5.2 --- METHODS / Chapter 5.2.1 --- Equipment --- p.42 / Chapter 5.2.2 --- Pilot Study for Reproducibility --- p.43 / Chapter 5.2.3 --- Estimation of Minimum Sample Size --- p.43 / Chapter 5.2.4 --- Morphometric Procedures --- p.44 / Chapter 5.2.5 --- Statistical Analysis --- p.48 / Chapter 5.2.6 --- Comparison of Visual Diagnosis of Cervical smears and biopsies --- p.49 / Chapter 5.2.7 --- Survey of Subjective Assessment Criteria for Cervical Biopsies and Smears --- p.50 / Chapter 6. --- RESULTS / Chapter 6.1 --- PILOT STUDY / Chapter 6.1.1 --- Intraobserver Reproducibility --- p.52 / Chapter 6.1.2 --- Minimum Sample Size --- p.52 / Chapter 6.2 --- CERVIX / Chapter 6.2.1 --- Maturation Sequence of Cervical Epithelium --- p.53 / Chapter 6.2.2 --- Differences of Morphometric Means between various groups of Cervical Biopsies --- p.56 / Chapter 6.2.3 --- Discriminant Analysis of Cervical Biopsies --- p.58 / Chapter 6.2.4 --- Differences of Morphometric Means between various groups of Cervical Smears --- p.60 / Chapter 6.2.5 --- Discriminant Analysis of Cervical Smears --- p.61 / Chapter 6.2.6 --- Comparison of Cervical Smears and Biopsies --- p.62 / Chapter 6.2.7 --- Subjective Assessment Criteria for Cervical Biopsies and Smears --- p.63 / Chapter 6.3 --- NASOPHARYNX / Chapter 6.3.1 --- Maturation Sequence of Nasopharyngeal Epithelium --- p.65 / Chapter 6.3.2 --- Differences of Morphometric Means between various groups of Nasopharyngeal Biopsies --- p.68 / Chapter 6.3.3 --- Discriminant Analysis of Nasopharyngeal Biopsies --- p.70 / Chapter 6.4 --- COMPARISON OF CERVIX UTERI AND NASOPHARYNX --- p.71 / Chapter 7. --- DISCUSSION / Chapter 7.1 --- CERVIX UTERI / Chapter 7.1.1 --- Maturation Sequence --- p.73 / Chapter 7.1.2 --- Discrimination of different groups in Biopsies --- p.76 / Chapter 7.1.3 --- Discrimination of different groups in Smears --- p.77 / Chapter 7.1.4 --- Comparison of Smears and Biopsies --- p.78 / Chapter 7.1.5 --- Subjective Assessment Criteria --- p.80 / Chapter 7.1.6 --- Future directions --- p.81 / Chapter 7.2 --- NASOPHARYNX / Chapter 7.2.1 --- Maturation Sequence --- p.81 / Chapter 7.2.2 --- Discrimination of different groups --- p.84 / Chapter 7.2.3 --- Nasopharyngeal Cytology --- p.84 / Chapter 7.2.4 --- Future directions --- p.85 / Chapter 7.3. --- COMPARISON OF CERVIX UTERI AND NASOPHARYNX / Chapter 7.3.1 --- Morphometric data --- p.85 / Chapter 7.3.2 --- Discriminant Analysis --- p.87 / Chapter 8. --- CONCLUSIONS --- p.89 / Chapter APPENDIX A --- Survey of subjective assessment criteria for cervical biopsies and smears / Tables A1-A7 --- p.92 / Chapter APPENDIX B --- Results of pilot study / Tables B1-B6 --- p.100 / Chapter APPENDIX C --- Morphometric data and results of statistical tests for cervical biopsies / Fig. C1-C61 --- p.104 / Tables C1-C19 --- p.166 / Chapter APPENDIX D --- Morphometric data and results of statistical tests for cervical smears / Fig. D1-D2 6 --- p.179 / Tables D1-D3 --- p.206 / Chapter APPENDIX E --- Comparison of cervical smears and biopsies / Tables E1-E3 --- p.208 / Chapter APPENDIX F --- Morphometric data and results of statistical tests for nasopharyngeal biopsies / Fig. F1-F61 --- p.211 / Tables F1-F12 --- p.273 / Chapter APPENDIX G --- Comparison of nasopharyngeal and cervical biopsies / Tables G1-G15 --- p.282 / Chapter APPENDIX H --- Pictures of materials and equipment / Fig. H1-H21 --- p.291 / REFERENCES --- p.303

by Cheng Man Kin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 71-76). / Abstracts in English and Chinese. / Thesis committee --- p.i / Abstract --- p.ii / Acknowledgements --- p.iv / Abbreviations --- p.v / Table of contents --- p.vii / List of figures --- p.x / List of tables --- p.xi / Chapter Chapter 1: --- General introduction --- p.1 / Chapter Chapter 2: --- Literature review --- p.3 / Chapter 2.1 --- Nutritional quality of plant proteins --- p.3 / Chapter 2.2 --- Using traditional plant breeding method to enhance amino acid quality of plant proteins --- p.3 / Chapter 2.3 --- Molecular strategies to enhance amino acid quality of plant proteins --- p.4 / Chapter 2.3.1 --- Heterologous gene expression --- p.5 / Chapter 2.3.2 --- Protein sequence modification --- p.8 / Chapter 2.3.3 --- Modification of biosynthesis pathway --- p.10 / Chapter 2.3.4 --- Synthetic gene expression --- p.11 / Chapter 2.3.5 --- Homologous gene overexpression --- p.13 / Chapter 2.4 --- Arabidopsis --- p.14 / Chapter 2.4.1 --- Arabidopsis as a model plant --- p.14 / Chapter 2.4.2 --- Transformation methods --- p.14 / Chapter 2.4.2.1 --- Direct DNA uptake --- p.15 / Chapter 2.4.2.2 --- Agrobacterium-mediated transformation --- p.15 / Chapter 2.5 --- Winged Bean Lysine-Rich protein --- p.17 / Chapter 2.5.1 --- Identification of winged bean polypeptides rich in lysine --- p.17 / Chapter 2.5.2 --- Cloning of the lysine-rich protein gene --- p.17 / Chapter 2.5.3 --- Further characterization of the WBLRP gene --- p.18 / Chapter 2.6 --- Phaseolin --- p.19 / Chapter Chapter 3: --- Expression of LRP in transgenic Arabidopsis --- p.20 / Chapter 3.1 --- Introduction --- p.20 / Chapter 3.2 --- Materials and methods --- p.21 / Chapter 3.2.1 --- Targeting LRP to cytosol --- p.21 / Chapter 3.2.1.1 --- Chemicals --- p.21 / Chapter 3.2.1.2 --- Plant materials --- p.21 / Chapter 3.2.1.3 --- Bacterial strains --- p.22 / Chapter 3.2.1.4 --- Construction of chimeric LRP gene (pBILRP-1) --- p.22 / Chapter 3.2.1.4.1 --- PCR amplification of LRP --- p.22 / Chapter 3.2.1.4.2 --- Cloning of PCR-amplified LRP into vector pD3-8 --- p.26 / Chapter 3.2.1.4.3 --- Cloning of recombinant plasmid pLRP-1 into binary vector --- p.26 / Chapter 3.2.1.5 --- Transformation of Agrobacterium with pBILRP-1 --- p.27 / Chapter 3.2.1.6 --- Vacuum infiltration transformation of Arabidopsis --- p.28 / Chapter 3.2.1.7 --- Selection of transgenic plants --- p.29 / Chapter 3.2.1.8 --- GUS assay --- p.30 / Chapter 3.2.1.9 --- DNA isolation --- p.31 / Chapter 3.2.1.10 --- PCR amplification and detection of transgenes --- p.31 / Chapter 3.2.1.11 --- Southern blot hybridization --- p.31 / Chapter 3.2.1.12 --- RNA isolation --- p.32 / Chapter 3.2.1.13 --- Reverse transcription-polymerase chain reaction (RT-PCR) --- p.32 / Chapter 3.2.1.14 --- Protein extraction and SDS-PAGE --- p.33 / Chapter 3.2.1.15 --- Protein sequencing --- p.33 / Chapter 3.2.1.16 --- Amino acid analysis --- p.34 / Chapter 3.2.2 --- Targeting LRP to protein bodies --- p.35 / Chapter 3.2.2.1 --- Chemicals --- p.35 / Chapter 3.2.2.2 --- Plant materials --- p.35 / Chapter 3.2.2.3 --- Bacterial strains --- p.35 / Chapter 3.2.2.4 --- Construction of chimeric LRP gene (pBILRP-2) --- p.35 / Chapter 3.2.2.4.1 --- Site-directed mutagenesis --- p.36 / Chapter 3.2.2.4.2 --- Cloning of the mutated phaseolin fragment into pBluescript --- p.36 / Chapter 3.2.2.4.3 --- PCR amplification of LRP --- p.39 / Chapter 3.2.2.4.4 --- Insertion of LRP into plasmid pBK/phas* --- p.39 / Chapter 3.2.2.4.5 --- Insertion of plasmid pLRP-2 into Agrobacterium binary vector --- p.41 / Chapter 3.2.2.5 --- Transformation of Agrobacterium with pBILRP-2 --- p.41 / Chapter 3.2.2.6 --- Vacuum infiltration transformation of Arabidopsis --- p.41 / Chapter 3.2.2.7 --- Selection of transgenic plants --- p.41 / Chapter 3.3 --- Results and discussion --- p.42 / Chapter 3.3.1 --- Targeting LRP to protein bodies --- p.42 / Chapter 3.3.1.1 --- Morphology of transgenic Arabidopsis --- p.42 / Chapter 3.3.1.2 --- Selection of transgenic plants --- p.42 / Chapter 3.3.2 --- Targeting LRP to cytosol --- p.46 / Chapter 3.3.2.1 --- Morphology of transgenic Arabidopsis --- p.46 / Chapter 3.3.2.2 --- Selection of transgenic plants --- p.46 / Chapter 3.3.2.3 --- Detection of GUS activity --- p.49 / Chapter 3.3.2.4 --- Integration of LRP transgene into Arabidopsis genome --- p.54 / Chapter 3.3.2.5 --- LRP transcript in transgenic Arabidopsis --- p.58 / Chapter 3.3.2.6 --- Stable accumulation of LRP in transgenic Arabidopsis --- p.61 / Chapter 3.3.2.7 --- Amino acid analysis of seed protein --- p.64 / Chapter Chapter 4: --- General discussion --- p.67 / Conclusion --- p.70 / References --- p.71

The main objective of this thesis is to take one step forward to practical and realisable devices for antenna and microwave engineering, using the technique of discrete coordinate transformation (DCT), which is a practical implementation of the coordinate transformation method. During this thesis, the DCT technique was demonstrated and analysed from the theory, and was proved to provide an all-dielectric approach to design devices under certain conditions. Two schemes were proposed on how to use this technique in a practical design. The first one is to transform an existing device into a flattened profile, meanwhile maintaining its electromagnetic performance. As examples, a flat reflector and a flat lens were created from a parabolic reflector and a convex lens, respectively. The second scheme is to project the propagating paths of an electromagnetic wave, and then generate a distorted space according to the paths by engineering the electromagnetic properties of the media. In this scheme, two examples of application were presented: an undetectable antenna composed of a carpet cloak and a conducting cavity, and a broadband device which can extraordinarily enhance the transmission through a sub-wavelength aperture. Numerical simulations based on the Finite-Difference Time-Domain (FDTD) method were implemented to verify all the designs. Several specific configurations were employed in the modelling in order to simulate the DCT based devices more efficiently and precisely. Performance of these devices was validated and analysed, and the advantages and disadvantages of this technique were investigated. Realisation and fabrication methods i were also studied, and a prototype was designed, fabricated and measured. At the end, as an extension, a multiple discrete coordinate transformation method was proposed and presented. This multiple transformation was proved to effectively relax the limitation of the one-step transformation, and was used to design an all-dielectric thin absorber from a conventional pyramidal one for demonstration.

This thesis is concerned with play, particularly the role of social play in a co-located context and its ability to bring people together. Participation in social play can have significant effects on an individual, group, community and culture, and thus, through practice-based research, this thesis documents the exploration of the design of “playful interventions” which may be artefacts or events which seek to bring people together through play. In play, individuals form shared meanings, understanding and values, as determined by the rules of the play situation. In the play experience, they become temporary communities, who, through play, can experiment, explore and redefine their relationships with one another, the play context and potentially the world beyond. The experimental nature of play leads it to be naturally imbued with transformative potential for everyone involved; whether that be small in scale, such as forming a new way of looking at a space through playing within it, or on a larger scale, through forming new concepts around a local area or governmental policy. Play is, however, very unpredictable, being led by player interaction, and always pushes up against the rules of the play situation. In play, the particular output (if there is one) is never certain, and no two play experiences will be the same. This unpredictability means that its transformational power is always a potential but never guaranteed. Designers, when working with play as a medium must embrace this unpredictability and explore approaches to design playful experiences which are satisfying in themselves for the participants whilst also trying to find methods to unlock the potential for individual (and group) transformation through play. The thesis is a narrative account of sustained academic research, based upon eight academic publications and practice works, produced between 2013 and 2018. Six of these publications document practical exploration of the creation of playful interventions, in the form of video games, performances and events. Two further publications explore design approaches to enhance participation drawing from expert interview analysis and theoretical engagement with institutional approaches to promotion of participation in the museum and gallery. The body of work thus explores the design of participation from two perspectives: the artist/designer of an artefact and as a “context provider” who designs events and spaces within which play, and participation can take place amongst participants. Within this thesis, the body of publications are contextualised in relation to theories of play, game design and art practice and also drawing from theories around communities of practice and communities of play. A series of expert practitioner interviews underpin both the academic and practical framing of this research, drawing from key practitioners in the UK and Europe working in play, game design, event curation and community work. The thesis formalises the design methods used to create playful interventions by the author and expert practitioners in the field of social play as presented both across the academic publications and within interview content. The formalisation of these design techniques is presented as two social play frameworks, one for designing participation around artefacts and one which focusses upon designing participation around events. Each framework aims to aid a designer and/or context provider in helping participants to unlock the unpredictable yet transformative potential of play as individuals and as communities whilst acknowledging the complex interrelations which occur in designed social contexts.

This study investigates how the subsidiaries of multinational enterprises (MNEs) reconfigure their resource bases to respond to an altered strategic positioning. In particular, the focus is on the subsidiaries of multinational enterprises from emerging economies (EMNEs), which undertake transformation from export-orientation to host market-orientation in an emerging economy being host country. Two Taiwanese MNEs with subsidiaries in China extensively operated the host market are selected as case studies. This research is grounded in a preliminary conceptualisation covering three main areas: subsidiary organisations, external actors in the host country and headquarters’ functions. It provides insights into (1) how the subsidiaries align their historical resources with newly-developed capabilities; (2) how the subsidiaries govern inter-firm relations with external actors in the host environment; and (3) whether and how complementary resources are to a limited extent transferred from headquarters to subsidiaries so as to support the operations in the host market. With regards to the organisational initiatives undertaken by the case subsidiaries, it is found that the historical resources are leveraged to support the host market business, whilst at the same time developing required marketing capabilities. In addition, the concept of organisational ambidexterity is adopted to refer to way in which existing (export-oriented) and new (host market-oriented) businesses that are operating simultaneously. As to the inter-firm relations, the case subsidiaries have been mainly governing their relations with distributors by performance-based mechanisms aiming to secure stable profits. By contrast, the relation-based trust was not commonly observed among the cooperation ties. With growing brand strengths and increased direct contact with consumers, they held higher authority over the interfirm relations with distributors than during the initial stages of operating in the Chinese market. In addition, with regards to the cross-border resource transfers, two distinct modes of headquarters-subsidiary relationships emerged: a traditional one, in which headquarters allocate resources within MNEs and a novel one in which headquarters’ functions were gradually replaced by the powerful subsidiary, termed in this thesis as “migrating headquarters”. On the basis of these findings, we put forward a set of propositions that present the interrelations between the resource circumstances of case subsidiaries, the institutional environments and the organisational initiatives undertaken by the case subsidiaries. Theoretically, the contributions of this study are threefold. Firstly, it advances the research on subsidiary development by holistically exploring the: resource reconfiguration of subsidiaries, inter-firm relations with external actors and headquarters-subsidiary relationships. In particular, the resource deficiency which the EMNEs’ subsidiaries encountered and the characteristics of the required capabilities for the host market-oriented transformation, i.e. local marketing competences, were investigated. Secondly, through probing the governance mechanisms adopted regarding interfirm relations between the case subsidiaries and local distributors, this study not only addresses the question of how MNEs acquire this location-bound resource, but also advances the extant research by the aspect of network positions. That is, this study indicates that the first-tier distributors hold more relations-based interactions with the case subsidiaries than the lower-tier ones did. Moreover, unlike the reliance on informal relations suggested by extant literature on doing business in emerging economies, it is found that the economic governance mechanisms based on distributor performance have been predominantly adopted by the case subsidiaries. Thirdly, by investigating how the complementary resources are transferred to the subsidiaries, this work discovers EMNEs’ weakness at responding to the host market-oriented subsidiary transformation in terms of resource deployment within MNEs, in particular those resources that have been mostly controlled by headquarters. In addition, the term “migrating headquarters”, which represents an extreme outcome of subsidiary development, provides novel knowledge to the extant literature on the relocation of MNE headquarters by the perspective of resource circumstances. Moreover, the five components comprising dynamic capabilities in the context of subsidiary transformation are identified through the two case studies as being: capability upgrading, capability leverage, capability building, coordination capability and cooperative capability.

My aim in this thesis is to tell the story/stories of how I arrived at a living theory of creativity which I shall call ‘knowledge transformation’. I explore this theory through ‘story’ as a methodology that connects both the creative writer and action researcher, and raises questions about self, reflective process and voice that are central to my enquiry. In telling these stories, I ask the question: what does it mean to be creative, as a writer, an educator and a manager? Is the nature of creativity transferable across each of these roles? How has this knowledge improved my practice as an educator? My examination leads to a theory of learning called ‘knowledge transformation’, which suggests that deep learning leads to change of both the learner and what is learnt. My premise is that ‘knowledge transformation’ involves the capacity to respond to challenge, self and other, and is central to the notion of creativity. I consider how far this capacity can be transferable, teachable and measurable in educational contexts, arriving at a notion of ‘scaffolded creativity’ which is demonstrated through practice in the higher academy. My journey towards and with this theory draws on my experience of four personae, the creative writer in and outside the academy, and the educator, team leader, and researcher within it; and explores the strategies and issues raised by bringing these roles and intelligences together. This theory of ‘knowledge transformation’ represents an aspirational contribution to our understanding of what it means to be ‘creative’. It explores how educational objectives can lead to deep learning and positive change. It also explores how values can be clarified in the course of their emergence and formed into living standards of judgment.

A linguagem C não provê segurança espacial de memória: não garante que a memória acessada através de um ponteiro para um objeto, tal como um vetor, de fato pertence ao objeto em questão. Em vez disso, o programador é responsável por gerenciar informações de alocações e limites, e garantir que apenas acessos válidos à memória são realizados pelo programa. Por um lado, isso provê flexibilidade: o programador tem controle total sobre o layout dos dados em memória, e sobre o momento em que verificações são realizadas. Por outro lado, essa é uma fonte frequente de erros e vulnerabilidades de segurança em programas C. Diversas técnicas já foram propostas para prover segurança de memória em C. Tipicamente tais sistemas mantêm suas próprias informações de limites e instrumentam o programa para garantir que a segurança de memória não seja violada. Isso causa uma série de inconvenientes, tais como mudanças no layout de memória de estruturas de dados, quebrando assim a compatibilidade binária com bibliotecas externas, e/ou um aumento no consumo de memória. Uma abordagem diferente consiste em usar tipos dependentes para descrever a informação de limites já latente em programas C e assim permitir que o compilador use essa informação para garantir a segurança espacial de memória. Embora tais sistemas tenham sido propostos no passado, eles estão atrelados especificamente à linguagem C. Outras linguagens, como C++, sofrem de problemas similares de segurança de memória, e portanto poderiam se beneficiar de uma abordagem mais independente de linguagem. Este trabalho propõe Týr, uma transformação de código baseada em tipos dependentes para garantir a segurança espacial de memória de programas C ao nível LLVM IR. O sistema permite que o programador descreva no nível dos tipos as relações entre pointeiros e informação de limites já presente em programas C. Dessa maneira, Týr provê segurança espacial de memória verificando o uso consistente desses metadados pré-existentes, através de verificações em tempo de execução inseridas no programa guiadas pela informação de tipos dependentes. Ao trabalhar no nível mais baixo do LLVM IR, Týr tem por objetivo ser usável como uma fundação para segurança espacial de memória que possa ser facilmente estendida no futuro para outras linguagens compiláveis para LLVM IR, tais como C++ e Objective C. Demonstramos que Týr é eficaz na proteção contra violações de segurança espacial de memória, com um overhead de tempo de execução relativamente baixo e de consumo de memória próximo de zero, atingindo assim um desempenho competitivo com outros sistemas para segurança espacial de memória de uma maneira mais independente de linguagem. / The C programming language does not enforce spatial memory safety: it does not ensure that memory accessed through a pointer to an object, such as an array, actually belongs to that object. Rather, the programmer is responsible for keeping track of allocations and bounds information and ensuring that only valid memory accesses are performed by the program. On the one hand, this provides flexibility: the programmer has full control over the layout of data in memory, and when checks are performed. On the other hand, this is a frequent source of bugs and security vulnerabilities in C programs. A number of techniques have been proposed to provide memory safety in C. Typically such systems keep their own bounds information and instrument the program to ensure that memory safety is not violated. This has a number of drawbacks, such as changing the memory layout of data structures and thus breaking binary compatibility with external libraries and/or increased memory usage. A different approach is to use dependent types to describe the bounds information already latent in C programs and thus allow the compiler to use that information to enforce spatial memory safety. Although such systems have been proposed before, they are tied specifically to the C programming language. Other languages such as C++ suffer from similar memory safety problems, and thus could benefit from a more language-agnostic approach. This work proposes Týr, a program transformation based on dependent types for ensuring spatial memory safety of C programs at the LLVM IR level. It allows programmers to describe at the type level the relationships between pointers and bounds information already present in C programs. In this way, Týr ensures spatial memory safety by checking the consistent usage of this pre-existing metadata, through run-time checks inserted in the program guided by the dependent type information. By targeting the lower LLVM IR level, Týr aims to be usable as a foundation for spatial memory which could be easily extended in the future to other languages that can be compiled to LLVM IR, such as C++ and Objective C. We show that Týr is effective at protecting against spatial memory safety violations, with a reasonably low execution time overhead and nearly zero memory consumption overhead, thus achieving performance competitive with other systems for spatial memory safety, in a more language-agnostic way.

The purpose of this thesis is to evaluate 7 different search strategies for content-based image retrieval with respect to retrieval effectiveness. The strategies are based on different levels of resolution implemented by the Haar transformation. The result of the study shows an improvement in both mean average precision and mean recall in strategies based on lower levels of resolution. / Uppsatsnivå: D

鈣粘蛋白是一類通過影響細胞粘附和細胞信號通路在腫瘤發生中起重要作用的細胞間粘附分子。鈣粘蛋白超家族包括經典鈣粘蛋白和非經典鈣粘蛋白，其中非經典鈣粘蛋白包含了原鈣粘蛋白。啟動子CpG甲基化調控下的基因沉默或表達下調是腫瘤發生中一個關鍵事件的觀點現已得到廣泛認可。一些鈣粘蛋白家族成員，如鈣粘蛋白-1/4/13（CDH1，CDH4，CDH13）被已有研究報導是受擬遺傳學調控沉默的功能性腫瘤抑制基因。本研究主要針對兩個鈣粘蛋白家族成員鈣粘蛋白-11（CDH11）和原鈣粘蛋白-10（PCDH10）進行腫瘤發生相關功能和機制的研究。 / CDH11位於雜合性缺失（LOH）經常發生而預示可能存在抑癌基因的染色體16q21-22區域，我們實驗室先前通過基因組芯片雜交技術（aCGH）對腫瘤細胞系的研究已發現它是該區域一個可能的抑癌基因。我們通過進一步的半定量反轉錄聚合酶鏈反應（RT-PCR）發現CDH11在正常組織和永生化正常上皮細胞中廣泛表達，但在各腫瘤細胞系中表達下降。甲基化特異性聚合酶鏈反應（MSP）和亞硫酸氫鹽處理的基因組測序（BGS）檢測到CDH11啟動子甲基化常發生于腫瘤細胞和腫瘤組織中。在CDH11表達缺失的腫瘤細胞中重新導入該基因的表達可顯著減少細胞克隆的形成，誘導細胞凋亡并抑制腫瘤細胞的遷移。通過更深入的機制研究，我們發現CDH11通過抑制Wnt/β-catenin信號通路發揮功能。 / 本研究的另一個鈣粘蛋白家族成員是PCDH10。之前我們實驗室的工作已經證實了PCDH10是一個在鼻咽癌和食管癌中受啟動子甲基化調控的抑癌基因,這裡我們主要研究它在大腸癌發病中的功能和機制。我們發現在PCDH10表達缺失的大腸癌細胞中重新導入PCDH10表達可顯著抑制腫瘤細胞的克隆形成，細胞遷移和幹細胞性。機制研究顯示PCDH10抑制Wnt/β-catenin和RhoA信號轉導通路，并進一步抑制腫瘤上皮細胞-間充質轉化（EMT）的過程，誘導幹細胞標記的下調。 / 綜上所述，本研究顯示CDH11和PCDH10兩個鈣粘蛋白家族成員在多種腫瘤中廣泛受甲基化調控失活，它們是重要的Wnt/β-catenin信號通路拮抗因素，可抑制腫瘤細胞的克隆形成和細胞遷移 / Cadherins are an important group of cell-cell adhesion molecules, which play crucial roles in tumorigenesis by affecting cell adhesion and cell signaling. Cadherin superfamily consists of classical cadherins and non-classical cadherins including protocadherins. It has been well recognized that silencing or downregulation of tumor suppressor genes (TSGs) by promoter CpG methylation is a critical event in human tumorigenesis. Some cadherin family members, such as CDH1, CDH4, CDH13, have been reported as functional TSGs silenced through epigenetic regulation. In this study, I mainly focus on the function and mechanism studies of two cadherin members-Cadherin 11(CDH11) and Protocadherin 10 (PCDH10). / CDH11 is located in 16q21-22, a region with frequent loss of heterozygosity (LOH), indicating the presence of candidate TSG. Previously, our lab also identified CDH11 as a candidate TSG through array-CGH of tumor cell lines. I further found by semi-quantitative RT-PCR that CDH11 was broadly expressed in normal tissues while frequently downregulated in multiple tumor cell lines, but not in immortalized normal epithelial cells. Methylation-specific PCR (MSP) and bisulfite genomic sequencing (BGS) detected frequent promoter methylation of CDH11 in tumor cell lines and primary tumor samples. Ectopic expression of CDH11 dramatically reduced tumor cell clonogenecity, induced tumor cell apoptosis and inhibited tumor cell migration. By further mechanism study, I found that CDH11 is a negative inhibitor of Wnt/β-catenin signaling pathway. / Another cadherin family protein which I chose to study is PCDH10. Previously our lab identified PCDH10 as a TSG by promoter methylation in nasopharyngeal and esophageal carcinomas. I studied the function and mechanism of PCDH10 in the pathogenesis of colon cancer. I found ectopic expression of PCDH10 strongly suppressed colon tumor cell clonogenecity, migration and stemness. Moreover, I found that PCDH10 repressed Wnt/β-catenin and RhoA signaling, thus further inhibited the epithelial-to-mesenchymal transition (EMT) of tumor cells and downregulated stem cell markers. / In summary, this study demonstrates two cadherin family members CDH11 and PCDH10, as important antagonists to Wnt/β-catenin signaling pathway, suppress tumor cell clonogenecity, migration, and are also frequently inactivated by epigenetic mechanism in multiple tumors. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Zhang, Yanjiao. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 84-95). / Abstracts also in Chinese. / Abstract --- p.i / Acknowledgements --- p.iii / Table of Contents --- p.vi / List of Tables --- p.x / List of Figures --- p.xi / List of Abbreviations --- p.xiii / List of Publications --- p.xvi / Chapter Chapter 1 --- Introduction and Literature Review --- p.1 / Chapter 1.1 --- Cancer --- p.1 / Chapter 1.1.1 --- General introduction about cancer --- p.1 / Chapter 1.1.2 --- Oncogenes and TSGs --- p.3 / Chapter 1.1.3 --- Cancer mechanism models --- p.3 / Chapter 1.2 --- Cancer Epigenetics --- p.5 / Chapter 1.2.1 --- DNA methylation --- p.6 / Chapter 1.2.2 --- DNA methylation and gene silencing --- p.7 / Chapter 1.2.3 --- DNA methylation and cancer --- p.7 / Chapter 1.2.4 --- Clinical implications of DNA methylation --- p.8 / Chapter 1.3 --- Cadherins --- p.10 / Chapter 1.3.1 --- General introduction of cadherin superfamily --- p.10 / Chapter 1.3.2 --- Cadherin classification --- p.10 / Chapter 1.3.3 --- Cadherin and cancers --- p.12 / Chapter 1.3.4 --- Cadherin switch and EMT in cancer --- p.16 / Chapter 1.4 --- Wnt/β-catenin signaling pathway and cancer --- p.16 / Chapter 1.4.1 --- Wnt/β-catenin signaling pathway --- p.16 / Chapter 1.4.2 --- Wnt/β-catenin signaling pathway in cancer --- p.18 / Chapter 1.4.3 --- Epigenetic silencing of Wnt/β-catenin signaling --- p.20 / Chapter 1.4.4 --- Wnt/β-catenin signaling pathway in CRC --- p.21 / Chapter Chapter 2 --- Aims of the Study --- p.22 / Chapter Chapter 3 --- Materials and Methods --- p.25 / Chapter 3.1 --- Cell lines and tissue samples --- p.25 / Chapter 3.1.1 --- Cell lines --- p.25 / Chapter 3.1.2 --- Maintenance of cell lines --- p.25 / Chapter 3.1.3 --- Drug treatment of cell lines --- p.26 / Chapter 3.1.4 --- Normal and primary tissues --- p.26 / Chapter 3.1.5 --- Total RNA extraction --- p.27 / Chapter 3.1.6 --- Genomic DNA extraction --- p.28 / Chapter 3.2 --- Gene expression analysis --- p.29 / Chapter 3.2.1 --- Reverse transcription (RT) --- p.29 / Chapter 3.2.2 --- Semi-quantitative RT-PCR --- p.30 / Chapter 3.3 --- Methylation Analysis --- p.32 / Chapter 3.3.1 --- CpG island prediction --- p.32 / Chapter 3.3.2 --- Sodium bisulfite treatment of genomic DNA --- p.33 / Chapter 3.3.3 --- Methylation-specific PCR (MSP) --- p.33 / Chapter 3.3.4 --- Bisulfite Genomic Sequencing (BGS) --- p.34 / Chapter 3.4 --- Construction of expression plasmids --- p.36 / Chapter 3.4.1 --- Construction of CDH11 expression vector --- p.36 / Chapter 3.4.2 --- Construction of PCDH10 expression plasmid --- p.38 / Chapter 3.4.3 --- Plasmid extraction --- p.39 / Chapter 3.5 --- Plasmid transfection --- p.41 / Chapter 3.6 --- Subcellular localization --- p.42 / Chapter 3.7 --- Function analyses --- p.43 / Chapter 3.7.1 --- Colony formation assay --- p.43 / Chapter 3.7.2 --- Wound healing assay --- p.44 / Chapter 3.8 --- Mechanism exploration --- p.44 / Chapter 3.8.1 --- Protein extraction and western-blot --- p.44 / Chapter 3.8.2 --- Dual-luciferase reporter assay --- p.47 / Chapter 3.9 --- Statistical analysis --- p.48 / Chapter Chapter 4 --- CDH11 functions as a tumor suppressor via modulating Wnt/β-catenin signaling and is frequently downregulated by promoter methylation --- p.49 / Chapter 4.1 --- The CpG island of CDH11 gene promoter --- p.50 / Chapter 4.2 --- CDH11 expression profile in normal tissues --- p.50 / Chapter 4.3 --- Frequent silencing of CDH11 by promoter methylation in multiple tumors --- p.51 / Chapter 4.4 --- Restoration of CDH11 expression by pharmacologic demethylation --- p.53 / Chapter 4.5 --- Frequent CDH11 methylation in primary tumors --- p.54 / Chapter 4.6 --- Function studies --- p.56 / Chapter 4.6.1 --- Ectopic expression of CDH11 inhibited tumor cell clonogenecity --- p.57 / Chapter 4.6.2 --- CDH11 induced tumor cell apoptosis --- p.57 / Chapter 4.6.3 --- CDH11 inhibited tumor cell migration --- p.58 / Chapter 4.7 --- CDH11 antagonized Wnt/β-catenin signaling --- p.59 / Chapter 4.8 --- Discussion --- p.60 / Chapter Chapter 5 --- Epigenetic inactivated tumor suppressor PCDH10 exerts tumor suppressive functions through modulating Wnt/β-catenin signaling and cell stemness in colon cancer --- p.66 / Chapter 5.1 --- PCDH10 was broadly expressed in normal tissues and frequently silenced in CRC cell lines --- p.67 / Chapter 5.2 --- Promoter methylation mediated PCDH10 silencing in CRC cell lines --- p.68 / Chapter 5.3 --- Frequent PCDH10 methylation in CRC primary tumors --- p.69 / Chapter 5.4 --- PCDH10 was located in cytoplasm and membrane --- p.70 / Chapter 5.5 --- Function Studies --- p.71 / Chapter 5.5.1 --- PCDH10 inhibited clonogenicity of tumor cells --- p.71 / Chapter 5.5.2 --- PCDH10 suppressed tumor cell mobility --- p.72 / Chapter 5.6 --- Mechanism exploration of PCDH10 in CRC --- p.72 / Chapter 5.6.1 --- PCDH10 antagonized Wnt/β-catenin signaling --- p.72 / Chapter 5.6.2 --- PCDH10 negatively regulated EMT and stemness of tumor cells --- p.74 / Chapter 5.6.3 --- PCDH10 inhibited RhoA signaling --- p.75 / Chapter 5.7 --- Discussion --- p.75 / Chapter Chapter 6 --- Conclusions and Future studies --- p.80 / Chapter 6.1 --- Conclusions --- p.80 / Chapter 6.2 --- Future studies --- p.82 / Reference List --- p.84