Wong Chi-Fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 119-132). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / List of Abbreviations --- p.v / List of Figures --- p.vii / List of Tables --- p.ix / Chapter 1 --- Introduction / Chapter 1.1 --- Plant secondary metabolites --- p.1 / Chapter 1.2 --- Plant alkaloids --- p.6 / Chapter 1.3 --- Narciclasine --- p.12 / Chapter 1.3.1 --- Isoquinoline alkaloids --- p.12 / Chapter 1.3.2 --- Amaryllidaceae alkaloids --- p.14 / Chapter 1.3.3 --- Narcissus --- p.16 / Chapter 1.3.4 --- Narciclasine --- p.17 / Chapter 1.3.4.1 --- Isolation --- p.17 / Chapter 1.3.4.2 --- Biological and pharmaceutical functions --- p.20 / Chapter 1.3.5 --- High performance liquid chromatography (HPLC) --- p.23 / Chapter 1.3.6 --- In vitro protein synthesis --- p.24 / Chapter 1.3.6.1 --- Rabbit reticulocyte lysate --- p.25 / Chapter 1.3.6.2 --- Wheat germ extract --- p.25 / Chapter 1.3.6.3 --- Non-radioactive colorimetric detection system --- p.26 / Chapter 1.4 --- Objective --- p.28 / Chapter 2 --- Materials and Methods / Chapter 2.1 --- Plant materials --- p.29 / Chapter 2.2 --- Extraction of narcicalsine --- p.29 / Chapter 2.3 --- Distribution of NCS in Narcissus tazetta --- p.30 / Chapter 2.4 --- Stability test --- p.31 / Chapter 2.4.1 --- HPLC analysis --- p.31 / Chapter 2.4.1.1 --- HPLC system --- p.31 / Chapter 2.4.1.2 --- Analytical condition --- p.31 / Chapter 2.4.2 --- Seed germination assay --- p.32 / Chapter 2.5 --- Mode of action of NCS --- p.33 / Chapter 2.5.1 --- In vitro translation --- p.33 / Chapter 2.5.1.1 --- In vitro translation --- p.33 / Chapter 2.5.1.2 --- SDS-PAGE analysis --- p.33 / Chapter 2.5.1.3 --- Western blot analysis --- p.34 / Chapter 2.5.1.4 --- Colorimetric detection --- p.34 / Chapter 2.5.2 --- Assay of induction of a-amylase synthesis in aleurone cells of barley grains by GA3 --- p.36 / Chapter 2.5.2.1 --- Chemicals and reagents --- p.36 / Chapter 2.5.2.2 --- Reducing sugar assay --- p.37 / Chapter 2.5.3 --- Root tip smear --- p.43 / Chapter 2.5.3.1 --- Chemicals and reagents --- p.43 / Chapter 2.5.3.2 --- Assay --- p.43 / Chapter 2.6 --- Allelopathic test --- p.45 / Chapter 2.6.1 --- Soil planting --- p.45 / Chapter 2.6.1.1 --- Foliage spray --- p.45 / Chapter 2.6.1.2 --- Planting with Narcissus bulb --- p.45 / Chapter 2.6.2 --- Hydroponics --- p.46 / Chapter 2.7 --- Effect of NCS on plant cells via tissue culture --- p.48 / Chapter 2.7.1 --- Establishment of tissue culture system --- p.48 / Chapter 2.7.1.1 --- Initiation and maintenance of carrot callus --- p.48 / Chapter 2.7.1.2 --- Initiation and maintenance of tobacco callus --- p.49 / Chapter 2.7.1.3 --- Initiation and maintenance of Narcissus callus --- p.50 / Chapter 2.7.1.4 --- Optimisation of callus growth --- p.50 / Chapter 2.7.2 --- Effects of NCS --- p.51 / Chapter 2.7.3 --- Effect of tobacco extract on NCS --- p.51 / Chapter 2.7.3.1 --- Extraction of tobacco extract --- p.51 / Chapter 2.7.3.2 --- Bioassay --- p.52 / Chapter 2.8 --- Assay of effect of NCS on microorganisms --- p.53 / Chapter 2.8.1 --- Antibacterial activity --- p.53 / Chapter 2.8.1.1 --- Total bacterial count --- p.53 / Chapter A. --- Chemicals and reagents --- p.53 / Chapter B. --- Serial dilution --- p.54 / Chapter C. --- Assay --- p.54 / Chapter 2.8.1.2 --- Turbidity test --- p.55 / Chapter A. --- Bacteria --- p.55 / Chapter B. --- Chemicals and reagents --- p.55 / Chapter C. --- Assay --- p.55 / Chapter 2.8.2 --- Anti-fungal and anti-yeast activity --- p.56 / Chapter 2.8.2.1 --- Disc diffusion method --- p.56 / Chapter A. --- Fungi --- p.56 / Chapter B. --- Chemicals and reagents --- p.56 / Chapter C. --- Assay --- p.56 / Chapter 2.8.2.2 --- Tube dilution method --- p.57 / Chapter A. --- Yeast --- p.57 / Chapter B. --- Chemicals and reagents --- p.57 / Chapter C. --- Assay --- p.57 / Chapter 2.9 --- Statistical analysis / Chapter 3 --- Results / Chapter 3.1 --- Distribution of NCS in Narcissus tazetta --- p.59 / Chapter 3.2 --- Stability of NCS --- p.62 / Chapter 3.2.1 --- HPLC analysis --- p.62 / Chapter 3.2.2 --- Bioassay --- p.62 / Chapter 3.3 --- Mode of action of NCS --- p.66 / Chapter 3.3.1 --- In vitro translation --- p.66 / Chapter 3.3.2 --- Effect ofNCS on the induction of a-amylase synthesis in aleurone cells of barley grains by GA3 --- p.69 / Chapter 3.3.3 --- Root tip smear --- p.74 / Chapter 3.4 --- Allelopathic test --- p.77 / Chapter 3.4.1 --- Soil planting --- p.77 / Chapter 3.4.1.1 --- Foliage applications --- p.77 / Chapter 3.4.1.2 --- Planting with Narcissus bulb --- p.77 / Chapter 3.4.2 --- Hydroponics --- p.78 / Chapter 3.5 --- Effect of NCS on plant cells via tissue culture --- p.91 / Chapter 3.5.1 --- Optimisation of Narcissus callus growth --- p.91 / Chapter 3.5.2 --- "Effects of NCS on Narcissus, carrot and tobacco calli" --- p.91 / Chapter 3.5.3 --- Effect of tobacco extract on NCS --- p.91 / Chapter 3.6 --- Effect of NCS on microorganisms --- p.95 / Chapter 3.6.1 --- Antibacterial activity --- p.95 / Chapter 3.6.1.1 --- Total bacterial count --- p.95 / Chapter 3.6.1.2 --- Turbidity test --- p.95 / Chapter 3.6.2 --- Anti-fungal and anti-yeast activities --- p.95 / Chapter 3.6.2.1 --- Disc diffusion method --- p.95 / Chapter 3.6.2.2 --- Tube dilution method --- p.96 / Chapter 4 --- Discussion / Chapter 4.1 --- General properties --- p.103 / Chapter 4.2 --- Mode of action --- p.105 / Chapter 4.3 --- Other biological properties --- p.108 / Chapter 4.3.1 --- Allelopathic property --- p.108 / Chapter 4.3.2 --- Effect on other plants via tissue culture --- p.110 / Chapter 4.3.3 --- Effect on microoraganisms --- p.112 / Chapter 4.4 --- Further studies --- p.114 / Chapter 5 --- Conclusion --- p.115 / Appendix --- p.116 / References --- p.119

by Cui Yanyan. / "June 1999." / Thesis (Ph.D.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (p. 154-163). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

No description available.

It was found that narciclasine retarded the growth of human cancer cells and plant suspension cells in dose-dependent manner. The inhibitory mechanism of narciclasine was found to be apoptosis for the DNA histogram showed an apoptotic peak in narciclasine-treated A375 cancer cells. The fluorescent signal dUTP fluorescein was found in the narciclasine-treated A735 cancer cell in TUNEL assay. The Annexin-V-FLUOS stained A375 cancer cell at 24-hour treatment with no PI found. These results suggest that narciclasine triggered early apoptosis in A375 cancer cell. Immunoblot analysis of the apoptotic signalling pathway showed that narciclasine induced apoptosis through the intrinsic pathway. Narciclasine induced the cleavage of caspase-9 but not the caspase-8, which was triggered by cytochrome c release from mitochondrial intermembrane space into cytosol. The activated caspase-9 triggered caspase cascade (e.g. cleavage of caspase-3, caspase-6 and caspase-7) which induced the cleavage of PARP. / Narciclasine is an isoquinoline alkaloid extracted from the bulb of Narcissus tazetta. It shows a wide range of biological activities such as antitumour, antiviral and plant growth inhibitory activities. However, little information is available regarding such inhibitory activities. The objective of this study is to elucidate the mechanisms of the cytotoxic effects of narciclasine in different cell models. / On the other hand, narciclasine triggered programmed cell death (PCD) in plant cells as proved by the increased intensity of Evans blue in narciclasine-treated suspension cells. Fluorescent microscopy showed that narciclasine induced PCD in tobacco BY2 cell with the dUTP fluorescein stained in narciclasine-treated cell. The induction of PCD was in dose-dependent and time-dependent manner. / Proteomic studies showed that narciclasine may affect A375 cancer cell and rice meristemic cells in similar manner. Narciclasine may affect the metabolism and defence system of both A375 cancer cell and rice meristemic cells through down-regulating the expression of metabolic enzymes (e.g. triosephosphate isomerase in A375 cancer cell and fructose bisphosphate aldolase in rice root tip) and defensive proteins (e.g. peroxiredoxin in A375 cancer cell and catalase in rice root tip). Narciclasine down-regulated the heat-shock proteins (HSP) which is involved in regulating cellular homeostasis and promoting cell survival. Therefore, narciclasine reduced HSP to lower the cell survival ability and induced the caspase cascade or caspase-like activity in A375 cancer cell and rice respectively. / To summarize, narciclasine induced apoptosis in A375 cancer cell and programmed cell death in tobacco BY2 cell. / Wong, Chi Fai. / "October 2007." / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4576. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 230-255). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.