Ha Wai-Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 166-180). / Abstracts in English and Chinese. / Acknowledgements --- p.ii / Abstract --- p.iii / Abbreviations --- p.vi / Table of Contents --- p.vii / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1 --- "Hstory, cultivation and trade" --- p.2 / Chapter 1.2 --- Botany --- p.4 / Chapter 1.3 --- Chemical Constituents and Pharmacological effects --- p.8 / Chapter 1.4 --- Authentication of Chinese herbal materials --- p.13 / Chapter 1.4.1 --- Morphological marker --- p.15 / Chapter 1.4.2 --- Histological marker --- p.18 / Chapter 1.4.3 --- Chemical marker --- p.20 / Chapter 1.4.4 --- Molecular markers --- p.24 / Chapter 1.4.4.1 --- Protein marker --- p.24 / Chapter 1.4.4.2 --- DNA-based markers --- p.26 / Chapter 1.4.4.2.1 --- PCR-based markers --- p.27 / Chapter 1.4.4.2.1.1 --- Random-primed PCR --- p.28 / Chapter 1.4.4.2.1.2 --- Simple Sequence Repeats (SSR) --- p.30 / Chapter 1.4.4.2.1.3 --- Polymerase Chain Reaction Fragment Length Polymorphism (PCR-RFLP) --- p.31 / Chapter 1.4.4.2.2 --- Hybridization-based markers --- p.33 / Chapter 1.4.4.2.3 --- Sequencing-based markers --- p.35 / Chapter 1.5 --- Objectives and Strategies of the studies --- p.39 / Chapter Chapter 2 --- General Materials and Methods --- p.40 / Chapter 2.1 --- Reagents and Buffers --- p.41 / Chapter 2.1.1 --- Media for bacterial culture --- p.41 / Chapter 2.1.2 --- Reagents for preparation of competent cells --- p.42 / Chapter 2.1.3 --- Reagents for plasmid DNA preparation --- p.42 / Chapter 2.1.4 --- Reagents for agarose gel electrophoresis --- p.43 / Chapter 2.1.5 --- Reagents for polyacrylamide gel electrophoresis --- p.43 / Chapter 2.1.6 --- Reagents for Southern hybridization --- p.44 / Chapter 2.2 --- Agarose Gel electrophoresis of DNA --- p.46 / Chapter 2.3 --- Purification of PCR products --- p.46 / Chapter 2.3.1 --- From agarose gel using Geneclean® II kit --- p.46 / Chapter 2.3.2 --- Using Microspin´ёØ Column --- p.47 / Chapter 2.4 --- End modification of PCR amplified DNA --- p.47 / Chapter 2.5 --- Preparation of Escherichia coli Competent Cells --- p.48 / Chapter 2.6 --- "Ligation and Transformation of E, coli" --- p.49 / Chapter 2.7 --- Plasmid Preparation --- p.50 / Chapter 2.7.1 --- Minipreparation of plasmid DNA --- p.50 / Chapter 2.7.2 --- Preparation of plasmid DNA using Wizard® Plus SV Minipreps DNA Purification Kit (Promega) --- p.50 / Chapter 2.8 --- Screening for the Presence of insert in plasmid --- p.51 / Chapter 2.8.1 --- Rapid alkaline lysis --- p.51 / Chapter 2.8.2 --- PCR screening --- p.52 / Chapter 2.8.3 --- Restriction digestion of plasmid DNA --- p.53 / Chapter 2.9 --- DNA sequencing --- p.53 / Chapter 2.9.1 --- Plasmid sequencing using T7 Sequencing Kit --- p.53 / Chapter 2.9.2 --- Cycle Sequencing from PCR products or plasmid --- p.54 / Chapter 2.10 --- DNA Sequencing electrophoresis --- p.55 / Chapter 2.10.1 --- Preparation of 6 % polyacrylamide gel solution --- p.55 / Chapter 2.10.2 --- Gel casting --- p.55 / Chapter 2.10.3 --- Electrophoresis of Sequencing Gel --- p.56 / Chapter 2.10.4 --- Autoradiography --- p.57 / Chapter 2.11 --- DNA elution from dried sequencing gel --- p.57 / Chapter 2.12 --- Southern blot analysis --- p.58 / Chapter 2.12.1 --- Restriction digestion of genomic DNA --- p.58 / Chapter 2.12.2 --- Purification of digested DNA and agarose gel electrophoresis --- p.58 / Chapter 2.12.3 --- Capillary transfer of DNA to a Hybond´ёØ N+ nylon membrane --- p.59 / Chapter 2.12.4 --- DNA radiolabeling by nick translation --- p.60 / Chapter 2.12.5 --- Purificaiton of radiolabeled probe by NICK® Spin Column --- p.60 / Chapter 2.12.6 --- Hybridization of DNA --- p.61 / Chapter Chapter 3 --- Plant DNA extraction --- p.62 / Chapter 3.1 --- Introduction --- p.63 / Chapter 3.2 --- Reagents and buffer for total DNA extraction --- p.66 / Chapter 3.3 --- Extraction methods --- p.70 / Chapter 3.3.1 --- Sample preparation --- p.70 / Chapter 3.3.2 --- CTAB extraction method --- p.70 / Chapter 3.3.3 --- Potassium acetate/ SDS extraction method --- p.71 / Chapter 3.3.4 --- GIBRO Plant DNAzol® reagent for genomic DNA isolation --- p.72 / Chapter 3.4 --- Qualitative and quantitative analysis of DNA --- p.74 / Chapter 3.5 --- Results --- p.75 / Chapter 3.6 --- Discussion --- p.78 / Chapter Chapter 4 --- Amplified Fragment Length Polymorphism (AFLP) analysis of P. ginseng and P. quinquefolius --- p.81 / Chapter 4.1 --- Introduction --- p.82 / Chapter 4.2 --- Materials and methods --- p.88 / Chapter 4.2.1 --- Plant materials --- p.88 / Chapter 4.2.2 --- Choice of Primers and radiolabeling --- p.89 / Chapter 4.2.3 --- AFLP assay --- p.90 / Chapter 4.2.4 --- Electrophoresis of AFLP fingerprint --- p.91 / Chapter 4.2.5 --- Similarity Index (S.I.) analysis of AFLP profile --- p.91 / Chapter 4.2.6 --- Re-amplification of polymorphic DNA fragments isolated from dried sequencing gel --- p.92 / Chapter 4.2.7 --- Cloning and Sequencing of the AFLP fragments --- p.93 / Chapter 4.2.8 --- Conversion of AFLP marker into Directed Amplification of Minisatellite-region DNA polymorphism (DAMD) marker --- p.93 / Chapter 4.3 --- Results --- p.95 / Chapter 4.4 --- Discussion --- p.102 / Chapter Chapter 5 --- Direct Amplification of Length Polymorphisms (DALP) analysis of P. ginseng and P. quinquefolius --- p.107 / Chapter 5.1 --- Introduction --- p.108 / Chapter 5.2 --- Materials and methods --- p.112 / Chapter 5.2.1 --- Plant materials --- p.112 / Chapter 5.2.2 --- Choice of Primers --- p.113 / Chapter 5.2.3 --- Alternative labelled Amplification reaction --- p.114 / Chapter 5.2.4 --- Electrophoresis of the multi-locus amplification products --- p.114 / Chapter 5.2.5 --- Isolation and Re-amplification of polymorphic DALP fragments from dried sequencing gel --- p.115 / Chapter 5.2.6 --- Cloning and Sequencing --- p.115 / Chapter 5.2.7 --- Conversion of DALP marker to Sequence Tagged Site (STS) marker --- p.116 / Chapter 5.3 --- Results --- p.117 / Chapter 5.4 --- Discussion --- p.135 / Chapter Chapter 6 --- Sequence-characterized amplified region (SCAR): the sequel of random amplified polymorphic DNA (RAPD) --- p.137 / Chapter 6.1 --- Introduction --- p.138 / Chapter 6.2 --- Materials and methods --- p.140 / Chapter 6.2.1 --- Plant materials --- p.140 / Chapter 6.2.2 --- PCR reaction --- p.141 / Chapter 6.2.3 --- Cloning and sequencing --- p.143 / Chapter 6.3 --- Results --- p.144 / Chapter 6.4 --- Discussion --- p.157 / Chapter Chapter 7 --- Outlook --- p.159 / Chapter 7.1 --- Molecular authentication of Chinese medicinal materials --- p.160 / Chapter 7.2 --- Development of molecular markers for Ginseng --- p.161 / Appendix I --- p.164 / Appendix II --- p.165 / References --- p.166

Wong Pak Ki. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 86-88). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / Dedication --- p.v / Table of Contents --- p.vi / List of Abbreviations --- p.ix / List of Appendices --- p.xi / List of Figures --- p.xiv / List of Tables --- p.xx / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Ginseng and Ginsenosides --- p.1 / Chapter 1.2 --- Instrumental Analysis of Ginsenosides --- p.6 / Chapter 1.2.1 --- Thin Layer Chromatography --- p.6 / Chapter 1.2.2 --- Infrared Spectroscopy --- p.7 / Chapter 1.2.3 --- Colorimetry --- p.7 / Chapter 1.2.4 --- Gas Chromatography --- p.7 / Chapter 1.2.5 --- High Performance Liquid Chromatography --- p.8 / Chapter 1.3 --- Objective of the Study --- p.9 / Chapter Chapter 2: --- Experimental --- p.13 / Chapter 2.1 --- History of Electrophoresis and Capillary Electrophoresis --- p.13 / Chapter 2.1.1 --- Electroosmotic Flow (EOF) --- p.14 / Chapter 2.1.2 --- Electrophoretic Migration --- p.18 / Chapter 2.2 --- Reagents and Materials --- p.20 / Chapter 2.2.1 --- Reagents and Glassware --- p.20 / Chapter 2.2.2 --- Instrumentation --- p.20 / Chapter 2.2.3 --- Preparation of Solutions and Wavelength Selection --- p.22 / Chapter 2.2 --- Procedures --- p.23 / Chapter Chapter 3: --- Results and Discussions --- p.24 / Chapter 3.1 --- Initial Selection of the Running Electrolyte --- p.24 / Chapter 3.2 --- Inclusion Additives in the Aqueous Buffer Solution --- p.29 / Chapter 3.2.1 --- Reasons for Addition of Buffer Additives --- p.29 / Chapter 3.2.1.1 --- Cyclodextrin --- p.29 / Chapter 3.3 --- Addition of Surfactants --- p.33 / Chapter 3.3.1 --- Sodium Dodecyl Sulfate (SDS) --- p.35 / Chapter 3.3.2 --- Sodium Cholate --- p.41 / Chapter 3.4 --- Addition of Organic Modifier --- p.43 / Chapter 3.5 --- Effect of pH --- p.46 / Chapter 3.6 --- Effect of the Concentration of the Borate/Phosphate Solution --- p.51 / Chapter 3.7 --- Effect of Capillaries with Different Inner Diameters (I.D.) --- p.54 / Chapter 3.7.1 --- Effect of pH --- p.54 / Chapter 3.7.2 --- Effect of the Buffer Concentration --- p.60 / Chapter 3.7.3 --- Comparison of Migration Time between Capillaries of 50μm and 75μm Inner Diameter --- p.62 / Chapter 3.8 --- Optimization of Other Experimental Parameters --- p.66 / Chapter 3.8.1 --- Applied Voltage --- p.66 / Chapter 3.8.2 --- The Time of Injection --- p.68 / Chapter 3.8.3 --- The Operating Temperature --- p.70 / Chapter 3.9 --- Intra-day and Inter-day Reproducibility --- p.72 / Chapter 3.10 --- Quantitative Analysis of the Ginsenosides --- p.74 / Chapter 3.11 --- Application of the Developed Methodology --- p.78 / Chapter 3.11.1 --- Experimental Procedures --- p.79 / Chapter Chapter 4: --- Conclusion --- p.83 / References --- p.86 / Appendices --- p.89

by Ngan Fai Ngor Karenda. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 128-134). / Acknowledgements --- p.i / Abstract --- p.ii / Table of Contents --- p.iii / Abbreviations --- p.viii / Chapter Chapter 1 --- Authentication of Chinese Medicinal Herbs / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Traditional Identification of Chinese Herbs / Chapter 1.2.1 --- Morphology --- p.3 / Chapter 1.2.2 --- Histology --- p.4 / Chapter 1.2.3 --- Chemical Analysis --- p.4 / Chapter 1.2.4 --- Proteins and Isozymes --- p.6 / Chapter 1.3 --- Molecular Technology in Authentication / Chapter 1.3.1 --- Restriction Fragment Length Polymorphism (RFLP) --- p.6 / Chapter 1.3.2 --- Polymerase Chain Reactions (PCRs) / Chapter 1.3.2.1 --- Random-Primed PCRs --- p.8 / Chapter 1.3.2.2 --- Simple Sequence Repeats --- p.10 / Chapter 1.3.2.3 --- Amplified Fragment Length Polymorphism (AFLP) --- p.11 / Chapter 1.4 --- Objectives and Strategies of the Study --- p.13 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Reagents and Buffers / Chapter 2.1.1 --- Buffers for Total DNA Extraction --- p.15 / Chapter 2.1.2 --- Reagents for Agarose Gel Electrophoresis --- p.16 / Chapter 2.1.3 --- Reagents for Polyacrylamide Gel Electrophoresis --- p.17 / Chapter 2.1.4 --- Reagents for Plasmid and Single-Stranded DNA Preparation --- p.17 / Chapter 2.1.5 --- Media for Bacterial Culture --- p.19 / Chapter 2.1.6 --- Reagents for Preparation of Competent Cells --- p.20 / Chapter 2.2 --- DNA Isolation / Chapter 2.2.1 --- Sample Preparation --- p.21 / Chapter 2.2.2 --- Cetyl triethylammonium bromide (CTAB) Extraction --- p.21 / Chapter 2.2.3 --- Cesium Chloride Gradient Ultracentrifugation --- p.21 / Chapter 2.3 --- Phenol/Chloroform Extraction --- p.22 / Chapter 2.4 --- Ethanol Precipitation --- p.23 / Chapter 2.5 --- DNA Concentration/Purity Estimation --- p.23 / Chapter 2.6 --- Random-Primed Polymerase Chain Reactions / Chapter 2.6.1 --- Random Amplified Polymorphic DNA (RAPD) --- p.24 / Chapter 2.6.2 --- Arbitarily-Primed Polymerase Chain Reaction (AP-PCR) --- p.24 / Chapter 2.7 --- rDNA Amplification --- p.24 / Chapter 2.8 --- Agarose Gel Electrophoresis of DNA --- p.25 / Chapter 2.9 --- Purification of rDNA / Chapter 2.9.1 --- from Agarose Gel using Geneclean II Kit (Bio 101 Inc.) --- p.25 / Chapter 2.9.2 --- using Microspin´ёØ Columns --- p.26 / Chapter 2.10 --- Preparation of Escherichia coli Competent Cells --- p.26 / Chapter 2.11 --- Ligation and Transformation of Escherichia coli --- p.27 / Chapter 2.12 --- Isolation of Plasmid DNA --- p.27 / Chapter 2.13 --- Screening of Plasmid DNA by Restriction Digestion --- p.28 / Chapter 2.14 --- Isolation of Plasmid DNA / Chapter 2.14.1 --- Minipreparation of Plasmid using Magic´ёØ Miniprep DNA Purification Kit from Promega --- p.28 / Chapter 2.14.2 --- Megapreparation of Plasmid using Qiagen-tip100 --- p.28 / Chapter 2.15 --- Single-Stranded DNA Preparation / Chapter 2.15.1 --- Transfection --- p.29 / Chapter 2.15.2 --- Single-Stranded DNA Isolation --- p.29 / Chapter 2.16 --- DNA Sequencing / Chapter 2.16.1 --- Plasmid Sequencing using T7 Sequencing Kit --- p.30 / Chapter 2.16.2 --- Cycle Sequencing from PCR Products --- p.30 / Chapter 2.16.3 --- Cycle Sequencing from PCR Products or Plasmid --- p.31 / Chapter 2.16.4 --- DNA Sequencing Electrophoresis --- p.31 / Chapter Chapter 3 --- Studies of Panax Species by Random-Primed PCRs / Chapter 3.1 --- Introduction --- p.34 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Plant Materials --- p.39 / Chapter 3.2.2 --- DNA Extraction and Random-Primed PCRs --- p.39 / Chapter 3.2.3 --- Data Analysis --- p.39 / Chapter 3.3 --- Results and Discussion / Chapter 3.3.1 --- DNA Isolation --- p.40 / Chapter 3.3.2 --- DNA Fingerprinting --- p.41 / Chapter 3.3.3 --- Relationship between the Six Panax Species --- p.45 / Chapter Chapter 4 --- Studies of Acorus by Random-Primed PCRs / Chapter 4.1 --- Introduction --- p.48 / Chapter 4.2 --- Materials and Methods / Chapter 4.2.1 --- Plant Materials --- p.49 / Chapter 4.2.2 --- DNA Extraction and Random-Primed PCRs --- p.50 / Chapter 4.3 --- Results and Discussion / Chapter 4.3.1 --- Acorus DNA --- p.50 / Chapter 4.3.2 --- Reproducibility of Random-Primed PCRs --- p.51 / Chapter 4.3.3 --- DNA Fingerprinting --- p.53 / Chapter Chapter 5 --- Studies of Epimedium by Random-Primed PCRs / Chapter 5.1 --- Introduction --- p.70 / Chapter 5.2 --- Materials and Methods / Chapter 5.2.1 --- Plant Materials --- p.71 / Chapter 5.2.2 --- DNA Extraction and Random-Primed PCRs --- p.71 / Chapter 5.3 --- Results and Discussion / Chapter 5.3.1 --- DNA Extraction --- p.71 / Chapter 5.3.2 --- DNA Fingerprinting --- p.72 / Chapter Chapter 6 --- Application of AP-PCR in Commercial Ginseng Products / Chapter 6.1 --- Introduction --- p.90 / Chapter 6.2 --- Materials and Methods / Chapter 6.2.1 --- Materials --- p.91 / Chapter 6.2.2 --- DNA Extraction and Random-Primed PCRs --- p.91 / Chapter 6.2.3. --- Data Analysis --- p.91 / Chapter 6.3 --- Results and Discussion / Chapter 6.3.1 --- DNA Isolation --- p.92 / Chapter 6.3.2 --- AP-PCR Analysis --- p.93 / Chapter Chapter 7 --- Ribosomal DNA as a Marker in Authentication of Panax Species / Chapter 7.1 --- Introduction --- p.99 / Chapter 7.2 --- Materials and Methods / Chapter 7.2.1 --- Plant Materials --- p.100 / Chapter 7.2.2 --- DNA Extraction and rDNA Amplification --- p.101 / Chapter 7.2.3 --- rDNA Sequencing --- p.101 / Chapter 7.2.4 --- Generation of Restriction Fragment Length Polymorphisms / Chapter 7.2.4.1 --- Restriction Digestion of rDNA Fragment --- p.102 / Chapter 7.2.4.2 --- Polyacrylamide Gel Electrophoresis (PAGE) --- p.103 / Chapter 7.2.4.3 --- Silver Staining for Nucleic Acids --- p.103 / Chapter 7.2.5 --- Data Analysis --- p.104 / Chapter 7.3 --- Results and Discussion / Chapter 7.3.1 --- rDNA Amplification and Plasmid Isolation --- p.104 / Chapter 7.3.2 --- rDNA Sequencing / Chapter 7.3.2.1 --- Sequence Comparison between the Six Panax species and the Two Adulterants --- p.107 / Chapter 7.3.3 --- Restriction Fragment Length Polymorphisms / Chapter 7.3.3.1 --- Restriction Profiles between Ginsengs and their Adulterants --- p.113 / Chapter 7.3.3.2 --- Restrciton Profiles of Ginsengs from Different Sources --- p.118 / Chapter 7.3.4 --- Panax Phylogeny --- p.121 / Chapter Chapter 8 --- General Discussion / Chapter 8.1 --- Advantages of Random-Primed PCRs --- p.124 / Chapter 8.2 --- Weaknesses of the Random-Primed PCRs --- p.125 / Chapter 8.3 --- Molecular Markers for Phylogenetic Studies --- p.126 / Chapter 8.4 --- Specific PCR-RFLP Patterns in Authentication --- p.126 / Chapter 8.5 --- Conclusions --- p.127 / References --- p.128 / Appendix --- p.135

No description available.

No description available.

The research undertaken in this thesis includes microwave-assisted extraction (MAE), synthesis, and the investigation of the scale-up of the microwave-assisted processes with the numerical aid. / The main goal of this research is to study the various problems associated with the scale-up of the microwave-assisted extraction and synthesis processes. Laboratory studies were carried out to investigate the microwave-assisted extraction of known components from peppermint leaves and American ginseng. Various factors that influence the extraction processes were studied. Microwave-assisted extraction method was compared with conventional heating and room temperature extraction methods on the extraction of ginsenosides from American ginseng. Microwave-assisted extraction method was determined to have higher extraction rate than both room temperature extraction and reflux temperature extraction using hotplate heating indicating that there is acceleration factor in enhancing the extraction rate beyond the temperature influence. / In the study of synthesizing n-butyl paraben, microwave-assisted synthesis was observed to greatly increase the yield of n-butyl paraben in much shorter period of time compared to the classic synthesis method. A transition state theory was proposed to explain this rate enhancement. The study of the synthesis of parabens with different alcohol and the influencing factors on the synthesis of n-butyl paraben yield were also studied. / A visualization method was developed to determine the microwave distribution in a domestic microwave cavity. The method uses gypsum plate as carrier and cobalt chloride as indictor. A simulation program was developed using the finite difference time domain (FDTD) approach and written in C programming language. The program was proved to be very versatile in different type of cavity simulation. Not only cavities with different dimensions and geometrical designs can be simulated, multiple magnetrons and various ways of magnetron placement can also be integrated into the simulation program. The detailed power distribution can be visualized in a 3-D plot, and the power distribution in each layer can be analyzed using the simulation result. The power distribution information will be very useful and necessary before any real equipment development.

Ginsenosides (GF) are a major bioactive constituent of ginseng and have been shown to elicit a multitude of actions ranging from the improvement of synaptic plasticity to the improved uptake of glucose into a cell. Furthermore, ginsenosides and their metabolites have been shown to be potent anti-cancer agents in multiple experimental cancer models. The aim of this study was to investigate the potential influence of GF derived from American ginseng root (Panax quinquefolius), and a ginsenoside metabolite Rh2, on tumor necrosis factor-α (TNF-α) cytotoxicity in MDA-MB 231 and MCF-7 human breast cancer cells. In combination, these agents significantly increased cell death in both cell lines. Together, ginsenosides and TNF-α induced a robust increase of the pre-G0/G1 and accompanying decrease in S phase cell populations in breast cancer cells. This cell death was the result of the induction of apoptosis, as determined by annexin-V/7-AAD and Hoechst staining. Furthermore, the mechanism of ginsenoside and TNF-α induced apoptosis is caspase-dependent, as determined by the pan-caspase inhibitor Z-VAD-FMK, with caspase-8, but not caspase-9, serving as initiator caspase in both cell lines. Additionally, ginsenoside treatment significantly XIAP expression in both MDA-MB 231 and MCF-7 cells, in the absence of TNF-α. In addition to enhancing apoptosis, it was also hypothesized that ginsenosides would abrogate pro-survival pathways induced by TNF-α. However, ginsenosides failed to block TNF-α effects on NFκB expression in either cell line. JNK which, when activated by TNF-α in MDA-MB 231 cells has a pro-survival function, was reduced by ginsenosides. However, JNK inhibition had no effect on cell death, suggesting that it does not play an integral role in the mechanism of action. In MCF-7 cells, JNK has been shown to have a pro-apoptotic function. Treatment with ginsenosides had no effect on TNF-α activation of JNK, but inhibition of JNK significantly reduced cell death in combined ginsenoside and TNF-α treated cells. To conclude, combined treatment with ginsenosides and TNF-α can enhance cell death in the sensitive MCF-7 cell line, and induce cell death in the insensitive MDA-MB 231 cell line in a caspase-dependent manner that is aided by the reduction of XIAP by ginsenosides.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2016-06-14T09:45:18Z No. of bitstreams: 1 texto completo.pdf: 3806262 bytes, checksum: 7171ac96932a9b0129a753982c0a71da (MD5) / Made available in DSpace on 2016-06-14T09:45:18Z (GMT). No. of bitstreams: 1 texto completo.pdf: 3806262 bytes, checksum: 7171ac96932a9b0129a753982c0a71da (MD5) Previous issue date: 2011-11-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo geral do presente estudo foi verificar a influência das trocas gasosas e do enriquecimento com CO 2 na propagação in vitro de Pfaffia glomerata (Spreng.) Pedersen (Amaranthaceae). Neste estudo foram utilizados como explantes segmentos nodais, a partir de culturas-estoque de vitroplantas de fáfia, mantidas sob subcultivos mensais. Foram avaliadas características relacionadas ao crescimento, aspectos anatômicos da folha, pigmentos fotossintéticos, teor de β-ecdisona (20E), atividade da Rubisco, amido, fenóis e açúcares. O primeiro capítulo teve como objetivo avaliar a influência da atmosfera ambiente e enriquecida com CO 2 (360 ou 720 μmol mol -1 de CO 2 ), vedação do recipiente de cultura e presença ou não de sacarose no meio de cultura durante o crescimento e desenvolvimento de explantes nodais de P. glomerata. Todas as características de crescimento das vitroplantas de fáfia aumentaram em condições de elevação de CO 2 . Nessas condições de atmosfera enriquecida com CO 2 foram produzidas vitroplantas de P. glomerata com características desejáveis para produção clonal massal, sendo uma alternativa para o estabelecimento de plantios comerciais que visem uniformidade e objetivem a produção de 20E para suprir a demanda industrial. A menor perda relativa de água das folhas oriundas de vitroplantas cultivadas em meio de cultura com ou sem sacarose e em condição de elevação de CO 2, mostra que um sistema fotoautotrófico ou fotomixotrófico com enriquecimento de CO 2 é atrativo para a aplicação na produção comercial massal de mudas dessa espécie, possivelmente reduzindo as perdas que ocorrem durante a aclimatização ex vitro, devido à desidratação das vitroplantas. O segundo capítulo teve como objetivo comparar a eficiência de novos tipos alternativos de membranas para vedação com as membranas MilliSeal ® , sobre a morfogênese e crescimento in vitro de fáfia. Dentre as membranas testadas, foi possível selecionar uma que mostrou características desejáveis para um sistema de propagação in vitro de plantas em larga escala, pois, aumentou o crescimento das vitroplantas de fáfia e apresenta custo unitário reduzido em comparação com membranas comercializadas atualmente. No terceiro capítulo é relatado o uso de substrato poroso combinado com o enriquecimento da atmosfera com CO 2 . Todas as características de crescimento das vitroplantas cultivadas em condições de elevação de CO 2 e em Florialite ® aumentaram. Em atmosfera enriquecida com CO 2 foram produzidas plantas de P. glomerata com alto acúmulo de biomassa e de 20E e apresentando alterações ultraestruturais. O presente estudo mostra que um sistema fotoautotrófico com enriquecimento de CO 2 pode ser atrativo para a aplicação na produção massal de mudas de fáfia ou ainda, para a produção de biomassa de fáfia com teor elevado de β-ecdisona. / The overall objective of this study was to investigate the influence of gas exchange and CO 2 enrichment on the in vitro propagation of Pfaffia glomerata (Spreng.) Pedersen (Amaranthaceae). Nodal segments from Pfaffia vitroplants maintained by monthly subculture were used as explants. Characteristics related to growth, leaf anatomy, photosynthetic pigments, β-ecdysone (20E) content, Rubisco activity, starch, phenols and sugars were evaluated. The objective of the first chapter was to evaluate the influence of the ambient atmosphere and the atmosphere enriched with CO 2 (360 or 720 μmol mol -1 CO 2 ), the closure system of culture vessels and presence or absence of sucrose in the culture medium during growth and development of Pfaffia nodal explants. All growth characteristics of Pfaffia vitroplants increased under high CO 2 . Under conditions of CO 2 -enriched atmosphere, P. glomerata vitroplants were produced with good characteristics for clonal mass production, which is an alternative for the establishment of homogeneous commercial plantations aiming to the production of 20E to meet the industrial demands. The low relative water loss from leaves of vitroplants grown in culture medium with or without sucrose and high CO 2 , shows that a photoautotrophic or photomixotrophic system with CO 2 enrichment is beneficial for mass production of seedlings of this species, possibly reducing losses during acclimatization due to dehydration of vitroplants. The second chapter aimed to compare the effectiveness of new alternative types of membranes for closure with the MilliSeal ® membrane on the growth and morphogenesis in vitro of Pfaffia. Among the membranes tested, the M3 showed the desirable characteristics for a system of large-scale in vitro propagation of plants, because it increased the growth of Pfaffia vitroplants and has reduced unit cost compared with currently marketed membranes. The third chapter reports on results obtained using porous substrates combined with atmosphere CO 2 enrichment. Vitroplants grown under high CO 2 and Florialite ® had all growth characteristics increased. Seedlings of P. glomerata produced in CO 2 -enriched atmosphere had high accumulation of biomass and 20E and showed ultrastructural alterations. The present study shows that a photoautotrophic system with CO 2 enrichment can be beneficial for either mass production of Pfaffia seedlings or the production of Pfaffia biomass with high 20E content. / Tese antinga

Made available in DSpace on 2014-06-11T19:25:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-27Bitstream added on 2014-06-13T20:13:52Z : No. of bitstreams: 1 tanimoto_a_me_botib_parcial.pdf: 743937 bytes, checksum: b5fbc721e900b87ed6ee516b188ac38b (MD5) Bitstreams deleted on 2015-06-25T13:01:00Z: tanimoto_a_me_botib_parcial.pdf,. Added 1 bitstream(s) on 2015-06-25T13:03:22Z : No. of bitstreams: 1 000719057_20160227.pdf: 666775 bytes, checksum: cc59d3dc12c5892bddd76906f22de973 (MD5) Bitstreams deleted on 2016-02-29T12:16:25Z: 000719057_20160227.pdf,. Added 1 bitstream(s) on 2016-02-29T12:17:24Z : No. of bitstreams: 1 000719057.pdf: 1458663 bytes, checksum: c74c3aef9b9308280cb043c348fb26a5 (MD5) / Pfaffia glomerata (Spreng.) Pedersen (“ginseng brasileiro”) é uma planta medicinal que apresenta um amplo espectro de usos como tônico, antiestresse e afrodisíaco. Esta espécie pertence ao complexo grupo de plantas medicinais conhecidas como “ginseng” e compõe um grupo específico de plantas denominadas adaptógenas, definidas como produtos capazes de aumentar a resistência não específica do organismo frente a diferentes estímulos estressores; considerando-se tal definição, é previsível que os compostos fitoquímicos presentes na P. glomerata sejam úteis no controle e prevenção de doenças na qual o estresse é fator etiológico, como é o caso da Doença Inflamatória Intestinal (DII). Dessa forma, o presente trabalho possui como objetivo avaliar a atividade anti-inflamatória intestinal da fração butanólica de P. glomerata (rica em saponinas), sobre o modelo de doença inflamatória intestinal induzida por ácido 2, 4, 6-trinitrobenzenosulfônico (TNBS) em ratos. Para tal, foram estudados parâmetros macroscópicos (escore da lesão, extensão da lesão, relação peso/comprimento do cólon, diarreia e aderência) e bioquímicos (atividade da mieloperoxidase e da fosfatase alcalina e quantificação da glutationa total) do processo inflamatório intestinal. Análises fitoquímicas mostraram principalmente a presença de triterpenos e saponinas, incluindo a ecdisterona, e indícios de compostos fenólicos. Os tratamentos com a fração butanólica de P. glomerata não foram capazes de impedir a formação lesões induzidas pelo TNBS e nem de auxiliar na resolução de um quadro inflamatório intestinal pré-existente, que também foi causado pelo TNBS. Com base nestes dados, a hipótese inicial de que um produto adaptógeno modularia o sistema imune por meio de sua ação antiestresse foi rejeitada nas condições experimentais utilizadas / Pfaffia glomerata (Spreng.) Perdersen, also called Brazilian ginseng, is a medicinal plant which has a broad use as tonic, anti-stress, and aphrodisiac. This specie belongs to the medicinal plants complex known as ginseng and it is part of a specific group of plants called adaptogens, which are defined as products that are capable of increasing non-specific body resistance against different stressors; considering the definition of adaptogens it is possible to assume that phytochemical compounds present in P. glomerata are also useful in controlling and preventing diseases in which stress is an etiological factor, such as Inflammatory Bowel Disease (IBD). Thereby, this present work aims to study the intestinal anti-inflammatory activity from the butanolic extract of P. glomerata (enriched in saponins) in the TNBS-induced inflammatory bowel disease in rats. To this, macroscopic (score, lesion extension, weight/length ratio, diarrhea and adherence) and biochemical (myeloperoxidase and alkaline phosphatase activities and total glutathione content) parameters of the inflammatory bowel disease were studied. Phytochemical analysis showed the presence of triterpens and saponins, including ecdysterone, and signs of phenolic compounds. Treatments with butanolic extract from P. glomerata were not able to prevent TNBS-induced lesions nor were able to ameliorate in the resolution of the pre-existing intestinal inflammatory process also caused by TNBS. According to these data, the initial hypothesis that an adaptogen product would modulate the immune system by its anti-stress action was rejected in the experimental conditions utilized

Made available in DSpace on 2014-06-11T19:32:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-06-24Bitstream added on 2014-06-13T19:03:03Z : No. of bitstreams: 1 alves_rbn_dr_botfca.pdf: 593371 bytes, checksum: 64b04d9d48af5b8086944eec1852a139 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O presente estudo teve como objetivos coletar germoplasma de Pfaffia glomerata (Spreng.) Pedersen, caracterizar genótipos provenientes de oito populações naturais, utilizando descritores morfológicos e um marcador químico e estabelecer um banco de germoplasma in vitro. Foram realizadas duas expedições de coleta, a primeira, à região localizada às margens do rio Santo Inácio e rio Tamanduá, Itatinga, SP e a segunda, ao Pantanal Sul Mato-grossense às margens do rio Paraguai, Corumbá, e às margens do rio Miranda, MS. Foram coletados 218 indivíduos em oito populações naturais que ainda não haviam sido amostradas e incorporação desses materiais ao banco de germoplasma in vitro da Unaerp, Ribeirão Preto, SP. Para a caracterização morfológica e química foi conduzido um experimento de campo com oito populações, em delineamento com blocos casualizados. As características avaliadas foram: altura da planta, comprimento do entrenó, diâmetro do caule, número de caules, tipo de caule, comprimento da raiz principal, número de raízes secundárias, diâmetro da raiz principal, matéria fresca e seca do caule, matéria fresca e seca das raízes, índice de colheita, a produtividade, cor caule, cor do pecíolo, pilosidade, cor da raiz, formato do limbo foliar, forma do ápice e da base do limbo foliar, relação comprimento/largura do limbo foliar, ocorrência de nematóides formadores de galhas e o teor de β-ecdisona. Foi registrada também a presença de insetos nas inflorescências. Os resultados foram submetidos à análise de variância, ao teste de separação de médias de Scott Knott a 5% de probabilidade. Visando definir o meio de cultura ideal para o estabelecimento do banco de germoplasma in vitro para a espécie foram avaliados seis tratamentos: 1) MS + 2% de sacarose + 4% de sorbitol; 2) MS/2 + 2% de sacarose + 4% de sorbitol; 3) MS + 2% de sacarose + 4% de sorbitol... / The aim in the present study was to collect germplasm of Pfaffia glomerata(Spreng.) Pedresen, characterize genotypes from eight natural populations utilizing morphological descriptors and a chemical marker and to establish a germplasm bank. The collecting expeditions were first to the region of the Santo Inacio and Tamandua riverbanks, Itatinga, SP and secondly to the Pantanal Sul region of the State of Mato Grosso, at the Paraguai and Miranda riverbanks. Two hundred and eighteen individuals were collected from eight natural populations not previously sampled and incorporated to the in vitro germplasm bank of the University of Ribeirao Preto (Unaerp), Ribeirao Preto Sao Paulo. For the morphological and chemical characterization the field experiment was conducted with eight populations and delineated in randomized blocks. The characteristics evaluated were: plant height, inter-nodule length, stem diameter, number of stems, type of stem, main root length, number of secondary roots, diameter of the main root, fresh and dry stem matter, fresh and dry root matter, stem color, petiole color, hairiness, root color, shape of the foliar limbus, shape of the apical part and base of the foliar limbus, length/width ratio of the foliar limbus, occurrence of gall forming nematodes, level of β-ecdysone and harvest index. Productivity and insect presence in the inflorescences was also verified. The results were submitted to analysis of variance and to the Scott Knott test of mean separations, with 5% probability. To define the best culture medium for the establishment of an in vitro germplasm bank six treatments were evaluated: 1) MS + 2% sacarose + 4% sorbitol; 2) MS/2 + 2% sacarose + 4% sorbitol; 3) MS + 2% sacarose + 4% sorbitol + 2mg.L-1 calcium pantothenate; 4) MS/2 + 2% sacarose + 4% sorbitol + 2mg.L-1 calcium pantothenate; 5) MS + 2% sacarose + 3% manitol + 2mg.L-1 calcium pantothenate; 6) ...(Complete abstract click electronic access below)