Typescript. / Thesis (Ph. D.)--University of Hawaii at Manoa, 1984. / Bibliography: leaves 114-123. / Photocopy. / Microfilm. / xi, 123 leaves, bound ill. (some col.) 29 cm

Typescript. / Thesis (Ph. D.)--University of Hawaii, 1968. / Bibliography: leaves [128]-132. / x, 132 l illus., tables

Numerous of studies have reported that polysaccharides have many bioactivities including immune system modulation, anti-oxidative and anti-tumor activities. Because herbal materials are rich in polysaccharides, some of the herbs like Dendrobium offcinale, its polysaccharide marker (DOP) has been applied in the quality analysis of polysaccharides in D. officinale by high performance gel permeation chromatography (HPGPC). However, such polysaccharide marker is not presented in every herb, and DOP also has limitation in monitoring herb formula. In addition, even though the same herbal materials and the same extraction methods were used, different studies gave out different conclusion regarding the structure of polysaccharides. Therefore, in the current study, D. officinale was used as a case study to demonstrate the application of ABEE-labeled oligosaccharides approach on structure elucidation and quantification of polysaccharides in herbs and herb formulae. First, the elucidation of polysaccharide structure remains challenging due to the lack of accurate analytical methods to determine the sequence and nature of glycosidic linkages. Oligosaccharide fragments from hydrolysis of polysaccharides are believed to provide accurate structure information, however, they are hard-to-separate and hard-to-detect. In the proposed method, the oligosaccharides generated from partial acid hydrolysis of DOP were labeled with p-aminobenzoic ethyl ester (ABEE), which made them separable and detectable by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS-DAD). Subsequently, nine ABEE-labeled oligosaccharide fragments (dimer to decamer) were isolated and identified by MS sequencing and 2D-NMR, and were confirmed by methylation analysis. The results indicated that the backbone should be β-D-1,4-linked Manp chain instead of mixed mannose and glucose chain. Second, two ABEE-labeled oligosaccharides namely, Te-Man-ABEE and Pen-Man-ABEE, were selected as chemical markers in the quantification of DOP in D. officinale and D.officinale (DO) products due to their high specificity in herb formula. The linear relationship between the content of these two markers and the content of DOP was successfully established. The linear relationship was further transformed to that between peak area of chemical markers and DOP content so that chemical markers were not necessary to be isolated for analysis. This linear relationship was systemically validated in terms of repeatability, precision and accuracy. The results showed that these two oligosaccharide markers presented a good linear relationship with DOP (R2 ≥ 0.997) in the range of 0.68-16.02 µg and also demonstrated satisfactory repeatability (RSD < 7.0%), and recovery (91.41% - 118.30%) in real sample determination. There was no significant difference between the results given by the two chemical markers as the RSD values were not more than 7.0%. While concerning the results given by the oligosaccharide-markers and the previously-published polysaccharide marker, the RSD value was not more than 6.4%. In conclusion, this approach provided an efficient and reliable method to obtain accurate structure information of polysaccharides and quantify specific polysaccharide in herb formula. it is believed that ABEE-labeled oligosaccharides approach can be also applied in the analysis of other saccharide-dominant herbal materials which in turns helps to find out the bioactivity mechanism of polysaccharides.

A multiplicação de orquídeas in vitro vem sendo utilizada há algum tempo com objetivo de elevar a taxa de multiplicação, além de eliminar patógenos e reduzir gastos na produção. Esta ferramenta de trabalho vem sendo rotineiramente utilizada no nosso laboratório, ao longo de mais de duas décadas em nosso laboratório, em pesquisas básicas de fisiologia e de aprimoramento da técnica de clonagem, principalmente de orquídeas. Neste caso,o uso da técnica visa a obtenção de maior estabilidade genética dos regenerantes em cultivos de longa duração. Plantas do gênero Catasetum apresentam atividade indeterminada do meristema apical caulinar quando incubadas no escuro, originando em pouco tempo longos estolões com crescimento indeterminado, comportamento raro no reino vegetal. Cada nó do caule estiolado possui uma gema lateral que, quando isolada e incubada no claro, forma rapidamente uma planta completa, facilitando a micropropagação. Outras espécies de orquídeas valorizadas na floricultura não apresentam tal facilidade na multiplicação, mostrando-se recalcitrantes à micropropagação, como é o caso do gênero Dendrobium (Orchidaceae). O objetivo deste estudo foi obter uma melhor compreensão dos mecanismos fisiológicos envolvidos no estiolamento de plantas Dendrobium \"Second Love\", que apresenta crescimento caulinar limitado quando sob ausência de luz, buscando compreender os efeitos do escuro e dos hormônios etileno e giberelina, bem como do radical livre óxido nítrico e gás carbônico na atividade dos meristemas apicais e laterais dessa orquídea. Como objetivo complementar, buscou-se estimular um estiolamento mais pronunciado, visando com isto um aumento potencial na formação de gemas laterais, paralelamente à quebra da dominância apical e o crescimento subsequente dos estolões. As plantas de Dendrobium utilizadas faziam parte do nosso estoque de germoplasma in vitro. Após 120 dias de incubação no claro, as plantas foram transferidas para o escuro e tratadas com diferentes concentrações de ácido giberélico (GA), paclobutrazol (PA - inibidor de biossíntese de giberelina), etileno, 1-metilciclopropeno (1-MCP - inibidor da ação do etileno) e óxido nítrico (NO). Análises mensais dos teores de etileno e CO2 acumulados nos frascos foram realizadas por meio de cromatografia gasosa durante três meses. Após 30, 60 e 90 dias de tratamento no escuro, quantificou-se o número de gemas laterais presentes nos estolões, o número de gemas laterais e apicais que se desenvolviam, o tamanho dos estolões formados, bem como os respectivos valores de massas fresca e seca destes. Por fim, buscou-se avaliar ainda a importância da incubação na penumbra e no escuro sobre o crescimento caulinar, o número de gemas laterais e o desenvolvimento destas após três meses de cultivo. O crescimento no escuro dos caules das plantas de Dendrobium \"Second Love\" mostrou-se extremamente lento e limitado quando comparado ao das plantas de Catasetum fimbriatum. No entanto, quando tratadas com 1.000 &mu;M de óxido nítrico, verificou-se ao final do terceiro mês que o número de gemas laterais era cinco vezes maior do que nas respectivas plantas controle. O tratamento com 10 ppm de etileno apresentou um aumento significativo no número de gemas e de estolões laterais, quando comparados ao controle a partir do segundo mês de incubação. Quanto ao tamanho do estolão apical, os tratamentos com 5 e 50 &mu;M de GA não apresentaram nenhum efeito promotor sobre alongamento caulinar. Mesmo não apresentando a retomada da atividade meristemática apical, o tratamento com 5 &mu;M de PA liberou um número maior de estolões laterais que o controle. Plantas tratadas com 1.000 ppm de NO, a partir do segundo mês de incubação, apresentaram um número elevado de estolões laterais, além dos mesmos apresentarem-se significativamente maiores. O tratamento com 100 ppm de 1-MCP apresentou o mesmo fenótipo das plantas tratadas no claro, ou seja, não estiolaram mesmo sob a ausência de luz. Quanto à emissão de etileno, observou-se que o tratamento com 1-MCP acarretou um aumento significativo na emissão deste gás pela planta, alcançando valores vinte vezes maiores do que no tratamento controle. Já a emissão de CO2 foi menor no tratamento claro quando comparada a maioria dos outros tratamentos no escuro. Os tratamentos em maiores concentrações de GA e NO pareceram promover algum tipo de estresse na planta (evidenciado pela necrose dos tecidos), demonstrando que a espécie em questão pode ser sensível à níveis elevados destas substâncias / In vitro multiplication has been used for some time in order to improve multiplication rate, eliminate pathogens and reduce the production costs. This working tool has been routinely used in our laboratory for over than two decades of basic research in plant physiology and enhancement of cloning technique, especially orchids, aiming to obtain greater genetic stability of regenerants in long term crops. Genus Catasetum present indeterminate shoot apical meristem activity when incubated in the dark, resulting, in a short period of time, long stolons with indeterminate growth: rare behavior in the plant kingdom. Each etiolated steam node has a lateral bud that, when isolated and incubated in light, quickly forms a complete plant, facilitating micropropagation. Other species of valued orchids in floriculture, such as genus Dendrobium (Orchidaceae), have no such facility in multiplication, being recalcitrant to micropropagation. The goals of this study were to gain a better understanding of the physiological mechanisms involved in plant etiolation in Dendrobium \"Second Love\", which has limited stem growth when in dark: and to understand the effects of dark, gibberellin and ethylene (plant hormones), as well as the free radical nitric oxide and carbon dioxide in the activity of apical and lateral meristems of the orchid. As a complementary objective, we tried to stimulate etiolation, aiming to potentially increase lateral buds formation and to break apical dominance with a subsequent stolons growth. Dendrobium plants used in this work were part of our in vitro germplasm stock. After 120 days of incubation in light, the plants were transferred to dark and treated with different concentrations of gibberellic acid (GA), paclobutrazol (PA - gibberellin biosynthesis inhibitor), ethylene, 1-methylcyclopropene (1-MCP - ethylene action inhibitor) and nitric oxide (NO). During a three months period, monthly analyzes of the accumulated levels of ethylene and CO2 in the flasks were performed using gas chromatography. After 30, 60 and 90 days of dark treatment the number of lateral buds presented in stolons, the number of developed lateral and apical buds, the size of formed stolons, and the respective amounts of fresh and dry mass were quantified. Finally, we evaluated the importance of incubation to steam growth in low light and in the dark, and the number of lateral buds and their development after three months of incubation. The Dendrobium \"Second Love\" steam growth in dark is extremely slow and limited when compared to Catasetum fimbriatum plants. However, after three months of treatment with 1.000 &mu;M nitric oxide it was found to have five times more lateral buds than the respective control treatment plants. The treatment with 10 ppm ethylene showed a significant increase in the number of buds and lateral stolons compared to the control treatment from the second month of incubation. Treatments with 5 and 50 &mu;M of GA had no promoting effect on the apical stolon stem elongation. Although not presenting the resumption of apical meristem activity, 5 &mu;M of PA treatment has released a greater number of lateral stolons than the control treatment. Plants treated with 1000 ppm of NO, from the second month of incubation, showed a higher number of lateral stolons, moreover they were significantly larger. Treatment with 100 ppm 1-MCP had the same phenotype as plants treated in light: in other words, they did not etiolate even in light absence. Regarding the ethylene emission, we observed that the treatment with 1-MCP caused a significant increase in the emission of this gas by the plant, reaching values twenty times higher than the control treatment. CO2 emission was lower in light treatment when compared to most of the other treatments in dark. Treatments at higher concentrations of NO and GA seemed to foster some sort of plant stress (evidenced by tissue necrosis), demonstrating that the specie in question may be sensitive to high levels of these substances

A multiplicação de orquídeas in vitro vem sendo utilizada há algum tempo com objetivo de elevar a taxa de multiplicação, além de eliminar patógenos e reduzir gastos na produção. Esta ferramenta de trabalho vem sendo rotineiramente utilizada no nosso laboratório, ao longo de mais de duas décadas em nosso laboratório, em pesquisas básicas de fisiologia e de aprimoramento da técnica de clonagem, principalmente de orquídeas. Neste caso,o uso da técnica visa a obtenção de maior estabilidade genética dos regenerantes em cultivos de longa duração. Plantas do gênero Catasetum apresentam atividade indeterminada do meristema apical caulinar quando incubadas no escuro, originando em pouco tempo longos estolões com crescimento indeterminado, comportamento raro no reino vegetal. Cada nó do caule estiolado possui uma gema lateral que, quando isolada e incubada no claro, forma rapidamente uma planta completa, facilitando a micropropagação. Outras espécies de orquídeas valorizadas na floricultura não apresentam tal facilidade na multiplicação, mostrando-se recalcitrantes à micropropagação, como é o caso do gênero Dendrobium (Orchidaceae). O objetivo deste estudo foi obter uma melhor compreensão dos mecanismos fisiológicos envolvidos no estiolamento de plantas Dendrobium \"Second Love\", que apresenta crescimento caulinar limitado quando sob ausência de luz, buscando compreender os efeitos do escuro e dos hormônios etileno e giberelina, bem como do radical livre óxido nítrico e gás carbônico na atividade dos meristemas apicais e laterais dessa orquídea. Como objetivo complementar, buscou-se estimular um estiolamento mais pronunciado, visando com isto um aumento potencial na formação de gemas laterais, paralelamente à quebra da dominância apical e o crescimento subsequente dos estolões. As plantas de Dendrobium utilizadas faziam parte do nosso estoque de germoplasma in vitro. Após 120 dias de incubação no claro, as plantas foram transferidas para o escuro e tratadas com diferentes concentrações de ácido giberélico (GA), paclobutrazol (PA - inibidor de biossíntese de giberelina), etileno, 1-metilciclopropeno (1-MCP - inibidor da ação do etileno) e óxido nítrico (NO). Análises mensais dos teores de etileno e CO2 acumulados nos frascos foram realizadas por meio de cromatografia gasosa durante três meses. Após 30, 60 e 90 dias de tratamento no escuro, quantificou-se o número de gemas laterais presentes nos estolões, o número de gemas laterais e apicais que se desenvolviam, o tamanho dos estolões formados, bem como os respectivos valores de massas fresca e seca destes. Por fim, buscou-se avaliar ainda a importância da incubação na penumbra e no escuro sobre o crescimento caulinar, o número de gemas laterais e o desenvolvimento destas após três meses de cultivo. O crescimento no escuro dos caules das plantas de Dendrobium \"Second Love\" mostrou-se extremamente lento e limitado quando comparado ao das plantas de Catasetum fimbriatum. No entanto, quando tratadas com 1.000 &mu;M de óxido nítrico, verificou-se ao final do terceiro mês que o número de gemas laterais era cinco vezes maior do que nas respectivas plantas controle. O tratamento com 10 ppm de etileno apresentou um aumento significativo no número de gemas e de estolões laterais, quando comparados ao controle a partir do segundo mês de incubação. Quanto ao tamanho do estolão apical, os tratamentos com 5 e 50 &mu;M de GA não apresentaram nenhum efeito promotor sobre alongamento caulinar. Mesmo não apresentando a retomada da atividade meristemática apical, o tratamento com 5 &mu;M de PA liberou um número maior de estolões laterais que o controle. Plantas tratadas com 1.000 ppm de NO, a partir do segundo mês de incubação, apresentaram um número elevado de estolões laterais, além dos mesmos apresentarem-se significativamente maiores. O tratamento com 100 ppm de 1-MCP apresentou o mesmo fenótipo das plantas tratadas no claro, ou seja, não estiolaram mesmo sob a ausência de luz. Quanto à emissão de etileno, observou-se que o tratamento com 1-MCP acarretou um aumento significativo na emissão deste gás pela planta, alcançando valores vinte vezes maiores do que no tratamento controle. Já a emissão de CO2 foi menor no tratamento claro quando comparada a maioria dos outros tratamentos no escuro. Os tratamentos em maiores concentrações de GA e NO pareceram promover algum tipo de estresse na planta (evidenciado pela necrose dos tecidos), demonstrando que a espécie em questão pode ser sensível à níveis elevados destas substâncias / In vitro multiplication has been used for some time in order to improve multiplication rate, eliminate pathogens and reduce the production costs. This working tool has been routinely used in our laboratory for over than two decades of basic research in plant physiology and enhancement of cloning technique, especially orchids, aiming to obtain greater genetic stability of regenerants in long term crops. Genus Catasetum present indeterminate shoot apical meristem activity when incubated in the dark, resulting, in a short period of time, long stolons with indeterminate growth: rare behavior in the plant kingdom. Each etiolated steam node has a lateral bud that, when isolated and incubated in light, quickly forms a complete plant, facilitating micropropagation. Other species of valued orchids in floriculture, such as genus Dendrobium (Orchidaceae), have no such facility in multiplication, being recalcitrant to micropropagation. The goals of this study were to gain a better understanding of the physiological mechanisms involved in plant etiolation in Dendrobium \"Second Love\", which has limited stem growth when in dark: and to understand the effects of dark, gibberellin and ethylene (plant hormones), as well as the free radical nitric oxide and carbon dioxide in the activity of apical and lateral meristems of the orchid. As a complementary objective, we tried to stimulate etiolation, aiming to potentially increase lateral buds formation and to break apical dominance with a subsequent stolons growth. Dendrobium plants used in this work were part of our in vitro germplasm stock. After 120 days of incubation in light, the plants were transferred to dark and treated with different concentrations of gibberellic acid (GA), paclobutrazol (PA - gibberellin biosynthesis inhibitor), ethylene, 1-methylcyclopropene (1-MCP - ethylene action inhibitor) and nitric oxide (NO). During a three months period, monthly analyzes of the accumulated levels of ethylene and CO2 in the flasks were performed using gas chromatography. After 30, 60 and 90 days of dark treatment the number of lateral buds presented in stolons, the number of developed lateral and apical buds, the size of formed stolons, and the respective amounts of fresh and dry mass were quantified. Finally, we evaluated the importance of incubation to steam growth in low light and in the dark, and the number of lateral buds and their development after three months of incubation. The Dendrobium \"Second Love\" steam growth in dark is extremely slow and limited when compared to Catasetum fimbriatum plants. However, after three months of treatment with 1.000 &mu;M nitric oxide it was found to have five times more lateral buds than the respective control treatment plants. The treatment with 10 ppm ethylene showed a significant increase in the number of buds and lateral stolons compared to the control treatment from the second month of incubation. Treatments with 5 and 50 &mu;M of GA had no promoting effect on the apical stolon stem elongation. Although not presenting the resumption of apical meristem activity, 5 &mu;M of PA treatment has released a greater number of lateral stolons than the control treatment. Plants treated with 1000 ppm of NO, from the second month of incubation, showed a higher number of lateral stolons, moreover they were significantly larger. Treatment with 100 ppm 1-MCP had the same phenotype as plants treated in light: in other words, they did not etiolate even in light absence. Regarding the ethylene emission, we observed that the treatment with 1-MCP caused a significant increase in the emission of this gas by the plant, reaching values twenty times higher than the control treatment. CO2 emission was lower in light treatment when compared to most of the other treatments in dark. Treatments at higher concentrations of NO and GA seemed to foster some sort of plant stress (evidenced by tissue necrosis), demonstrating that the specie in question may be sensitive to high levels of these substances

Sj?gren’s syndrome (SS) is a chronic autoimmune disorder of the exocrine glands with clinical manifestation of dry eyes and dry mouth. The pathogenesis of SS is poorly understood, and efficient therapy is currently lacking. Therefore, an appropriate animal model recapitulating the key features of SS could be of profound value. Although several murine models have been established and evaluated, some of these models may develop other non-SS symptoms simultaneously. Herein, an autoimmunization-induced C57BL/6 female mouse model is evaluated. This mouse model exhibited less saliva secretion and swollen salivary gland with severe inflammation in the submandibular gland. Furthermore, apoptosis and pro-inflammatory cytokines were significantly increased and the expression of aquaporin 5, a water channel protein, was decreased and restricted to the basolateral membranes in acinar cells, indicating the weakening of water transport. Besides, autoantibodies such as Ro, La and other anti-nuclear autoantigens were found to be tremendously increased. The expression of M3 muscarinic receptor (M3R) increased in the acinar cells. This can be described as a compensatory expression due to the long term blockage from the autoantibodies which is similar in the SS patients. The characteristics of this model recapitulate the key features of human SS. Dendrobium officinale is an herbal medicine with yin-nourishing effect and clinically used in China as a health tonic to promote body fluid production. The polysaccharides extracted from Dendrobium officinale (DP) consisted of 6 monosaccharides: mannose, glucose, galactose, arabinose, xylose and glucuronic acid in the ratio of 10:0.25:1.2:4.7:1.3:1.4. DP was found to be protective on this experimental SS model. Specifically, administration of DP 20 mg/ml significantly prevented the chaos of immune responses and imbalance of pro-inflammatory cytokines. Our previous work also demonstrated that DP can promote saliva production in both SS patients and SS model. Therefore, we investigated the M3R activation induced by DP treatment. In contrast to the acute activation by acetylcholine, DP exerts slow, but long term activation on M3R. The botanical monosaccharides xylose and arabinose cannot be detected in the cell lysate, indicating that hydrolyzed DP did not permeate the cell membrane. Further investigations suggested that DP can inhibit the apoptosis induced by the addition of recombinant TNF-α on the human salivary gland epithelial cell line A-253. By investigating the potential mechanisms, we found that DP treatment can inhibit the apoptotic factors including the NF-κB activation, increased reactive oxygen species, decreased mitochondrial membrane potential and prolonged mitogen-activated protein kinase activation. The results suggested that DP may interfere with the TNF-α pathway and its receptor since DP did not permeate the cell membrane. / published_or_final_version / Chinese Medicine / Doctoral / Doctor of Philosophy

Flowering time and flower quality of three nobile dendrobium hybrids in relation to vernalization duration and light intensity during vernalization were studied in the first experiment. Mature Dendrobium Red Emperor 'Prince', Dendrobium Sea Mary 'Snow King', and Dendrobium Love Memory 'Fizz' were cooled at 10 degrees C with 300 to 350 mol·m–2·s–1 photosynthetic photon flux (PPF) (12-h photoperiod) or darkness, each with four cooling durations (2, 4, 6, or 8 weeks). Plants were forced in a greenhouse after vernalization. At least 4 weeks of 10 degrees C cooling in light was needed for flower initiation of Red Emperor 'Prince'; whereas Sea Mary 'Snow King' and Love Memory 'Fizz' only needed 2 weeks of 10 degrees C cooling regardless of light. Darkness during vernalization slightly delayed flowering and resulted in fewer but larger flowers. Longer cooling duration delayed flowering, decreased the flower longevity, and produced larger and more flowers. In the second experiment, Love Memory 'Fizz' were cooled at 15 degrees C for 4 weeks with PPF of 0, 50, 100, or 200 mol·m–2·s–1 (12-h photoperiod). Compared to 200 mol·m–2·s–1, low PPF of 50 or 100 mol·m–2·s–1 did not affect flowering time or flower quality; however, darkness delayed flowering and reduced flower quality. The third experiment was aimed at developing a strategy to defer flowering of nobile dendrobium orchids by holding them under low temperature. Mature Den. Red Emperor 'Prince' and Den. Sea Mary 'Snow King' were held at 10 degrees C for various durations (0, 4, 8, 12 or 16 weeks) after vernalization (4 weeks at 10 degrees C). Plants were forced in a greenhouse after holding. Time to flowering, flower differentiation and flower quality were determined. Increase of low temperature holding duration from 0 to 16 weeks extended time to flowering up to 3 months and did not affect parameters of flower except producing larger flowers and reducing flower number per flowering node for Den. Red Emperor 'Prince'. Notably, the flower longevity was not adversely affected. Defoliation was aggravated in Den. Red Emperor 'Prince' by longer duration of cooling and was considered a detrimental effect of low temperature holding.

by Luk Choi Wan. / Thesis submitted in: November 2005. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 125-132). / Abstracts in English and Chinese. / Abstract --- p.iv / 摘要 --- p.vi / Acknowledgements --- p.viii / Tabel of contents --- p.x / List of Figures --- p.xii / List of Tables --- p.xiv / List of Abbreviations --- p.xv / List of Abbreviations --- p.xv / Chapter Chapter One --- Literature review --- p.1 / Chapter 1.1 --- General introduction --- p.1 / Chapter 1.2 --- General aspects of plant lectins --- p.4 / Chapter 1.3 --- Monocot mannose-binding lectins --- p.5 / Chapter 1.3.1 --- General introduction --- p.5 / Chapter 1.3.2 --- Sugar specificity --- p.6 / Chapter 1.3.3 --- Isolation and purification --- p.7 / Chapter 1.3.4 --- Molecular cloning --- p.9 / Chapter 1.3.5 --- Molecular structure and modifications --- p.10 / Chapter 1.3.6 --- Molecular evolution --- p.12 / Chapter 1.3.7 --- Transformation --- p.12 / Chapter 1.3.8 --- Physiological roles --- p.14 / Chapter 1.3.9 --- Application --- p.19 / Chapter 1.4 --- Dendrobium nobile --- p.31 / Chapter 1.4.1 --- Background --- p.31 / Chapter 1.4.2 --- Chemical analysis --- p.32 / Chapter Chapter Two --- Biological activities of a mannose-binding lectin isolated from Dendrobium mobile --- p.33 / Chapter 2.1 --- Introduction --- p.33 / Chapter 2.2 --- Materials and methods --- p.35 / Chapter 2.2.1 --- Mannose-binding lectin from D. nobile --- p.35 / Chapter 2.3 --- Biological activities of mannose-binding lectin from D. nobile --- p.36 / Chapter 2.3.1 --- Hemagglutinating activity --- p.36 / Chapter 2.3.2 --- In vitro anti-proliferative assay --- p.37 / Chapter 2.3.3 --- In vitro antiviral assay --- p.40 / Chapter 2.3.4 --- Statistical Analysis --- p.42 / Chapter 2.4 --- Results --- p.43 / Chapter 2.4.1 --- Physiochemical properties of D. nobile lectins --- p.43 / Chapter 2.4.2 --- Cytotoxicity to cancer cell lines --- p.53 / Chapter 2.4.3 --- Antiviral activity --- p.59 / Chapter 2.5 --- Discussion --- p.60 / Chapter Chapter Three --- Molecular cloning of lectin gene of Dendrobium nobile --- p.65 / Chapter 3.1 --- Introduction --- p.65 / Chapter 3.2 --- Methods --- p.67 / Chapter 3.2.1 --- RNA extraction --- p.67 / Chapter 3.2.2 --- RT-PCR synthesis of D. nobile lectin cDNA --- p.68 / Chapter 3.2.3 --- RACE of D. nobile agglutinin gene --- p.69 / Chapter 3.2.4 --- Generation of DNL full-length lectin gene sequence --- p.70 / Chapter 3.2.5 --- Cloning and sequencing of PCR product --- p.71 / Chapter 3.2.6 --- Data analyses --- p.72 / Chapter 3.2.7 --- Synthesis of single-stranded DIG-labeled DNA probe --- p.74 / Chapter 3.2.8 --- Northern blot analysis --- p.74 / Chapter 3.2.9 --- Genomic DNA extraction --- p.75 / Chapter 3.2.10 --- Southern blot analysis --- p.76 / Chapter 3.2.11 --- Expression of DNL in E. coli --- p.76 / Chapter 3.2.12 --- Western blot analysis --- p.78 / Chapter 3.3 --- Results --- p.80 / Chapter 3.3.1 --- Isolation and characterization of DNL gene --- p.80 / Chapter 3.3.2 --- Sequence analysis of DNL --- p.89 / Chapter 3.2.3 --- Secondary and tertiary structure --- p.96 / Chapter 3.2.4 --- Southern blot analysis --- p.99 / Chapter 3.2.5 --- Northern blot analysis --- p.99 / Chapter 3.2.6 --- Expression of fusion protein in E.coli --- p.102 / Chapter 3.4 --- Discussion --- p.104 / Chapter Chapter Four --- General Discussion --- p.110 / Chapter 4.1 --- General discussion --- p.110 / Chapter 4.2 --- Isolation and Characterization of monocot mannose-binding lectin of D. nobile --- p.111 / Chapter 4.3 --- Molecular cloning of monocot mannose-binding lectin of D. nobile --- p.115 / Chapter 4.4 --- Further investigations --- p.122 / Chapter Chapter Five --- Conclusion --- p.124 / References: --- p.125 / Appendix --- p.133

by Zhang Yanbo. / "December 2003." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (p. 163-185). / 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.

Thesis (M.Sc. (Plant Science))--Mahidol University, 2006. / LICL has E-Thesis 0018 ; please contact computer services.