So, Cheung Wai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 106-117). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.iii / 摘要 --- p.v / List of Abbreviations --- p.vii / List of Figures --- p.viii / List of Tables --- p.x / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Candida albicans and candidiasis / Chapter 1.1.1 --- Historical background --- p.1 / Chapter 1.1.2 --- C. albicans infections --- p.2 / Chapter 1.1.3 --- Current challenges in the treatment of C. albicans --- p.3 / Chapter 1.2 --- Photodynamic therapy --- p.11 / Chapter 1.2.1 --- Historical aspects and development --- p.11 / Chapter 1.2.2 --- Basic principle of photodynamic therapy --- p.13 / Chapter 1.2.3 --- Light applicator --- p.16 / Chapter 1.2.4 --- Generations of photosensitizer --- p.17 / Chapter 1.2.5 --- Characteristics of phthalocyanines --- p.20 / Chapter 1.2.6 --- Photodynamic antimicrobial chemotherapy (PACT) --- p.22 / Chapter 1.3 --- Aim of present study --- p.24 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Synthesis of BAM-SiPc --- p.27 / Chapter 2.2 --- Preparation of BAM-SiPc solution --- p.27 / Chapter 2.3 --- Yeast strains and culture conditions --- p.28 / Chapter 2.4 --- Light source --- p.29 / Chapter 2.5 --- Assays of PDT with planktonic C. albicans / Chapter 2.5.1 --- Photodynamic treatment on planktonic cells --- p.30 / Chapter 2.5.2 --- Clonogenic assay --- p.32 / Chapter 2.5.3 --- Cellular uptake of BAM-SiPc --- p.32 / Chapter 2.5.4 --- Distribution of BAM-SiPc in planktonic cells / Chapter 2.5.4.1 --- Fluorescence microscopic analyses --- p.33 / Chapter 2.5.4.2 --- Confocal laser scanning microscopic (CLSM) analyses --- p.34 / Chapter 2.5.5 --- Determination of ROS level in planktonic cells --- p.34 / Chapter 2.5.6 --- Distribution of ROS in planktonic cells --- p.35 / Chapter 2.5.7 --- Effect of ROS inhibitors --- p.35 / Chapter 2.5.8 --- Membrane integrity assay --- p.36 / Chapter 2.6 --- Assays of PDT with C. albicans biofilm / Chapter 2.6.1 --- Biofilm formation --- p.37 / Chapter 2.6.2 --- Photodynamic treatment on C. albicans biofilm --- p.38 / Chapter 2.6.3 --- Viability assays / Chapter 2.6.3.1 --- XTT reduction assay --- p.38 / Chapter 2.6.3.2 --- Molecular probes staining --- p.40 / Chapter 2.6.4 --- Determination of ROS level in biofilm --- p.41 / Chapter 2.6.5 --- Distribution of BAM-SiPc in biofilm --- p.41 / Chapter 2.6.6 --- Photodynamic treatment on C. albicans from resuspended biofilm --- p.42 / Chapter 2.7 --- Statistical analysis --- p.42 / Chapter Chapter 3 --- Results / Chapter 3.1 --- BAM-SiPc mediated PDT on planktonic C. albicans / Chapter 3.1.1 --- Antifungal effect of BAM-SiPc on C. albicans / Chapter 3.1.1.1 --- PDT activities on different strains of C. albicans --- p.43 / Chapter 3.1.1.2 --- Effect of different densities of cells --- p.47 / Chapter 3.1.1.3 --- Effect of a washing step before illumination --- p.47 / Chapter 3.1.2 --- Optimization of PDT conditions with BAM-SiPc on C. albicans / Chapter 3.1.2.1 --- Time course study --- p.50 / Chapter 3.1.2.2 --- Light dose study --- p.50 / Chapter 3.1.3 --- Uptake of BAM-SiPc --- p.53 / Chapter 3.1.4 --- Distribution of BAM-SiPc in the planktonic cells / Chapter 3.1.4.1 --- Analysis with fluorescence microscopy --- p.56 / Chapter 3.1.4.2 --- Analysis with CLSM --- p.56 / Chapter 3.1.5 --- ROS production upon PDT treatment / Chapter 3.1.5.1 --- ROS level in the planktonic cells --- p.59 / Chapter 3.1.5.2 --- Distribution of ROS production --- p.61 / Chapter 3.1.5.3 --- Effect of different ROS inhibitors on BAM-SiPc's potency --- p.64 / Chapter 3.1.6 --- Membrane integrity --- p.66 / Chapter 3.2 --- BAM-SiPc mediated PDT on C. albicans biofilm / Chapter 3.2.1 --- Establishment of the biofilm model with 192887g --- p.69 / Chapter 3.2.2 --- Photodynamic treatment on 192887g biofilm / Chapter 3.2.2.1 --- Viability assay - XTT assay --- p.72 / Chapter 3.2.2.2 --- Viability assay ´ؤ LIVE/DEAD BacLight Bacterial Viability kit --- p.72 / Chapter 3.2.3 --- ROS level in the biofilm after PDT treatment --- p.75 / Chapter 3.2.4 --- Distribution of BAM-SiPc in the biofilm --- p.75 / Chapter 3.2.5 --- Photodynamic treatment on C. albicans from resuspended biofilm --- p.79 / Chapter Chapter 4 --- Discussion --- p.81 / Chapter 4.1 --- Antifungal effect of BAM-SiPc on the planktonic C. albicans --- p.81 / Chapter 4.2 --- Effects of different conditions on the photodynamic treatment with BAM- SiPc --- p.83 / Chapter 4.3 --- Mechanistic study of the antifungal effect of BAM-SiPc --- p.86 / Chapter 4.3.1 --- Interaction between BAM-SiPc and C. albicans --- p.86 / Chapter 4.3.2 --- ROS as mediator of cell damage --- p.89 / Chapter 4.3.3 --- Analysis of membrane integrity upon photodynamic treatment --- p.91 / Chapter 4.4 --- Establishment of the biofilm model of C. albicans --- p.92 / Chapter 4.5 --- In vitro effect of BAM-SiPc mediated PDT on C. albicans biofilm --- p.95 / Chapter Chapter 5 --- Conclusion and Future Perspectives / Chapter 5.1 --- Conclusion --- p.101 / Chapter 5.2 --- Future perspectives --- p.102 / References --- p.106
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326528 |
| Date | January 2008 |
| Contributors | So, Cheung Wai., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, x, 117 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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