Ampelopsis grossedentata is a wildly used herb in South China as sleep aid beverage for many years. Yet the active ingredients and mechanisms of this herb were unknown. In the present study, extract from Ampelopsis grossedentata which we named ECBRC-AG, and one of its active ingredient myricetin were proved having significant hypnotic/sedative effects in multiple animal models. ECBRC-AG shortened sleep latency, increase NREM sleep and decrease locomotor activity when treated before the onset of light period in rats. ECBRC-AG could decrease active awake and increase REM sleep in the late part of light period. ECBRC-AG also decreased the caffeine induced hyperactivity in rats. Among the three suspected active ingredients from ECBRC-AG, myricetin showed similar active profile with ECBRC-AG. Myricetin increased NREM and REM sleep, decreased sleep latency, decreased locomotor activity and also active awake. All the above evidences have implicated that myricetin is the most important active ingredient of ECBRC-AG ECBRC-AG and myricetin did not show any obvious side effects on rats. / Based on these findings, we propose that myricetin facilitates GABA function on PVN neurons through a T-type calcium channel and CaM-KII mechanism. The hypnotic/sedative effects of ECBRC-AG and myricetin are mediated by PVN. ECBRC-AG treatment decreased corticosterone levels in rats, which also indicated that PVN/HPA axis was the target of these herbal derivates. PVN has broad interactions with GABAergic, hypocretinergic, cytokine and NPY system and all these systems are proved to be deeply involved in sleep regulation. / In conclusion, the present study has identified that myricetin is the most important active ingredient of the herbal extract ECBRC-AG. We confirmed the hypnotic/sedative effects of ECBRC-AG and myricetin on rats, and also revealed the different action profiles of these herbal derivates compared with zolpidem. T-type calcium channels and the HPA axis were shown to be involved in the mechanisms of ECBRC-AG and myricetin, indicating that they may be the new targets for insomnia treatment with these herbal derivates. / Insomnia is the most common sleep disorder and affects about one third of the general population. Insomnia is always combined with physical and mental illness, as either a consequence or a contributing factor. Insomnia produces sleepiness, impairment in psychomotor performance, absenteeism, frequent accidents, memory impairment and a high risk of depression. Pharmacologic therapies are the most important interventions for insomnia. However, the currently available hypnotics are associated with residual effects and risks of abuse and dependence. More efficient and safe hypnotics are needed. / The DNA array and RT-PCR studies revealed that GABA, hypocretin, cytokine and NPY systems were involved in the mechanisms of ECBRC-AG and myricetin. In calcium imaging study, we found that myricetin induced a transient Ca 2+ influx in the primary culture of rat hypothalamus neurons. This Ca2+ influx could be blocked by T-type channel blocker mibefradil. RT-PCR study also showed that ECBRC-AG and myricetin treatment changed the mRNA expression level of T-type calcium channel al G subunit in rat hypothalamus. The present results are consistent with our previous study showing that myricetin enhanced GABA function in the neurons of rat hypothalamic paraventricular nucleus (PVN), and that blocking CaM-KII pathway eliminated this effect. / Zhang, Xiaohu. / "March 2008." / Adviser: Chan Hsiao Chang. / Source: Dissertation Abstracts International, Volume: 70-03, Section: B, page: 1516. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (p. 156-174). / 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.

Orientador: Luciane Dias de Oliveira / Banca: Antonio Olavo Cardoso Jorge / Banca: Marcos Roberto Furlan / Resumo: Visando o potencial terapêutico do extrato glicólico de B. pendula em tratamentos de infecções bacterianas e fúngicas e de doenças inflamatórias, foram avaliadas suas atividades antimicrobiana, antiinflamatória, citotóxica e genotóxica. A atividade antimicrobiana do extrato foi analisada em Candida albicans, C. dubliniensis, C. glabrata, C. guilliermondii, C. krusei, C. tropicalis, Staphylococcus aureus, Streptococcus mutans, Escherichia coli e Pseudomonas aeruginosa com a determinação das Concentrações Inibitória Mínima e Microbicida Mínima (CIM e CMM) em culturas planctônicas e posteriormente testado sobre biofilmes monotípicos. A atividade citotóxica foi avaliada em cultura de macrófagos de camundongo (RAW 264.7) e fibroblastos gengivais humanos (FMM-1) após exposição de 5 min e 24 h ao extrato, com o teste MTT e teste imunoenzimático (ELISA) que quantificou a produção das citocinas IL-1β e TNF-α produzidas por RAW 264.7. A genotoxicidade do extrato foi avaliada pelo teste de micronúcleos. A atividade antiinflamatória foi avaliada nos sobrenadantes coletados de culturas de RAW 264.7 estimuladas com LPS de E. coli e expostas aos extratos (5 min e 24 h) pela quantificação de citocinas IL-1β, TNF-α e IL-10 por ELISA e óxido nítrico (ON) pelo método de Griess. A análise estatística foi realizada por ANOVA e teste de Tukey, com significância de 5%. O extrato teve ação antimicrobiana em todos biofilmes, com redução acima de 39,5% em 5 min e acima de 78% em 24 h. A viabilidade celular foi satisfatória em concentrações abaixo de 50 mg/mL em 5 min tanto para FMM-1, quanto para RAW 264.7, contudo em 24 h concentrações acima de 3,13 e 12,5 mg/mL foram citotóxicas para RAW 264.7 e FMM-1, respectivamente. Não houve indícios de ação genotóxica do extrato. A atividade anti-inflamatória foi evidenciada pela redução significativa na produção de TNF-a e ON nos grupos tratados . Conclui-se que o... / Abstract: In order to verify the therapeutic potential of glicolic extract of B. pendula on the threatment of bacterial and fungical infection and on inflammatory diseases, its antimicrobial, anti-inflammatory actions, cytotoxic and genotoxic effects were evaluated. The antimicrobial activity of the extracts was tested on Candida albicans, C. dubliniensis, C. glabrata, C. guilliermondii, C. krusei, C. tropicalis, Staphylococcus aureus, Streptococcus mutans, Escherichia coli and Pseudomonas aeruginosa with determination of Minimum Inhibitory and Minimum Microbicide Concentrations (MIC and MMC) in planktonic growth, following of test in monotypic biofilms. The cytotoxic activity was evaluated in mouse macrophages (RAW 264.7) and human gingival fibroblast (FMM-1) after exposure of 5 min and 24 h to extract, through the MTT test and by Enzyme Linked Immunosorbent Assay (ELISA) to quantify the production of the cytokines IL-1β and TNF-α by RAW 264.7. The genotoxicity of the extracts was evaluated by micronucleus test. The anti-inflammatory activity was evaluated in RAW 264.7 stimulated with LPS from E. coli, after the period of exposure to the extract (5 min and 24 h) the supernatant was removed to quantify pro-inflammatory (IL-1β and TNF-α.) and antiinflammatory (IL-10) cytokines by ELISA and nitric oxide (NO) by Griess method. Statistical analysis was performed by ANOVA and Tukey's test, with significance level of 5%. The extract has shown antimicrobial activity with reduction above of 39.5% of biofilm in 5 min and more than 78% in 24 h. The cell viability was satisfactory at concentrations below 50 mg/mL in 5 min to RAW 264.7 and FMM-1, however in 24 h concentrations above 3.13 and 12.5 mg/mL were cytotoxic to RAW 264.7 and FMM-1, respectively. There was no evidence of genotoxic action of the extract. The anti-inflammatory activity was evidenced by the significant reduction in the production o... / Mestre

Orientador: Luciane Dias de Oliveira / Banca: Luis Felipe das Chagas e Silva de Carvalho / Banca: Jônatas Rafael de Oliveira / Resumo: As plantas medicinais e os fitoterápicos têm sido utilizados como coadjuvantes e alternativos no combate a diversas doenças com crescente frequência, no entanto, na Odontologia seu uso ainda é bastante limitado. Com isso, o objetivo deste estudo é avaliar a ação antimicrobiana dos extratos aquoso e glicólico de própolis verde sobre micro-organismos anaeróbios de interesse odontológico, bem como sua citotoxicidade, a fim de introduzir e incentivar o uso efetivo e sistemático desse fitoterápico em produtos como dentifrícios e enxaguatórios bucais no combate a cáries e doenças periodontais. Os extratos comerciais aquoso e glicólico de própolis foram obtidos das empresas Apis Flora e Mapric, respectivamente. Para avaliação da atividade antimicrobiana foram utilizadas cepas-padrão (ATCC) de Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis Porphyromonas gingivalis e Prevotella intermedia em cultura planctônica, verificando a concentração inibitória mínima e concentração microbicida mínima (CIM e CMM), segundo Clinical and Laboratory Standards Institute. Para biofilmes monotípicos, suspensões padronizadas (107 céls/mL) foram adicionadas em poços de microplacas e após 48 h em anaerobiose foram tratados com 3 concentrações do extrato de própolis (n=12) por 5 min. Foi incluído um controle positivo (solução fisiológica) e um controle negativo (clorexidina). O biofilme foi mensurado pelos testes MTT e Cristal Violeta. Para análise de citotoxicidade, queratinócitos hu... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Medicinal plants and herbal medicines have been used as adjuvants and alternatives in fighting various diseases with increasing frequency. However, in dentistry its use is still quite limited. Therefore, the objective of this study is to evaluate the antimicrobial action of the aqueous and glycolic extracts of green propolis on anaerobic microorganisms of dental interest, in order to introduce and encourage the effective and systematic use of this herbal medicine in products such as dentifrices and mouthwashes in the fight against tooth decay and periodontal diseases. Aqueous and glycolic commercial extracts of propolis were obtained from Apis Flora and Mapric companies, respectively.To evaluate the antimicrobial activity, standard strains (ATCC) of Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella intermedia in planktonic culture was used, verifying the minimum inhibitory concentration and minimum microbicide concentration (MIC and CMM), according to Clinical and Laboratory Standards Institute. For monotypic biofilms, standardized suspensions (107 cells / mL) was added to microplate wells and after 48 h in anaerobiosis was treated with 3 concentrations of propolis extracts (n = 12) for 5 min. A positive control (saline solution) and a negative control (chlorhexidine) was included. The biofilm was measured by the MTT and Violet Crystal tests. For cytotoxicity analysis, human keratinocytes (HaCaT) were cultured and ... (Complete abstract click electronic access below) / Mestre

O chá verde (Camellia sinensis L) apresenta potencial de uso em dietas dos animais domésticos devido à grande quantidade de polifenóis presentes em suas folhas. Pesquisas desenvolvidas com animais, realizadas primeiramente com cobaias, mostram resultados positivos na nutrição e comportamento animal. O objetivo deste estudo foi avaliar o efeito da adição de extrato de chá verde à dieta de novilhas leiteiras sobre o comportamento, consumo de concentrado e desenvolvimento corporal. Foram utilizadas no experimento 32 novilhas não gestantes. O delineamento experimental utilizado foi o de blocos completos casualizados, com medidas repetidas no tempo e quatro tratamentos. As variáveis avaliadas foram: consumo de concentrado e matéria seca, ganho de peso, medidas corporais, comportamento ingestivo junto ao cocho e medidas fisiológicas. O consumo de concentrado foi maior (P<0,05) nas doses zero e 3g que nas doses de 1 e 2g de extrato de chá verde. O ganho de peso dos animais que receberam 2 e 3g/d de extrato de chá verde tendeu a ser maior (P<0,10) que o dos animais que receberam 1g ou o controle (zero). A inclusão de extrato de chá verde na dieta de novilhas não influenciou o consumo de matéria seca total, eficiência alimentar, as medidas corporais, comportamentais e fisiológicas, mas influenciou o consumo de concentrado e tendeu a influenciar o ganho de peso diário. / Green tea (Camellia sinensis L) is potentially useful as a dietary additive for domestic animals due to the polyphenols found in the leaves. The aim of this study was to evaluate the addition of green tea extract into the concentrate of dairy heifers on behavior, concentrate intake and body development. Thirty-two Holstein and Jersey dairy heifers not pregnant were selected. The experimental design was a randomized complete block design with repeated measures and four treatments: zero, 1, 2 and 3 grams of green tea extract added into the concentrate. Variants used: concentrated intake and dry matter, weight gain, body measurements, ingestion behavior along with trough and physiological dimension. The concentrated intake is higher (P<0,05) in doses zero and 3g than in doses with 1 and 2 g of green tea extract. The weight gain of the subjects that received 2 and 3 g/d of green tea extract tended to be higher (P<- 0,10) than those that received 1 g or the control sample (zero). The inclusion of green tea extract in the diet of heifers didn’t affect the ingestion of total dry matter, nurture efficiency nor the bodily, behavioural and the physiological measurements, but it did influence the concentrated intake and trended to sway the daily weight gain.

Sun Huimin. / Thesis submitted in: December 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 77-89). / Abstracts in English and Chinese. / TITLE PAGE --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABSTRACT --- p.iii / ABSTRACT IN CHINESE --- p.v / LIST OF PUBLICATIONS --- p.vii / ABBREVIATIONS --- p.viii / TABLE OF CONTENTS --- p.ix / Chapter CHAPTER 1. --- General Introduction --- p.1 / Chapter 1.1 --- Current Status of Herbal Product Use --- p.1 / Chapter 1.2 --- Herb-drug interactions --- p.2 / Chapter 1.2.1. --- Mechanisms of herb-drug interaction --- p.3 / Chapter 1.2.2. --- Pharmacodynamic interaction --- p.3 / Chapter 1.2.3. --- Pharmacokinetic interaction --- p.4 / Chapter 1.2.4. --- Herb-drug interaction involving drug metabolizing enzymes --- p.5 / Chapter 1.3 --- Methodologies for studying herb-drug interactions involving CYP enzymes --- p.7 / Chapter 1.3.1. --- Animal studies (Ex vivo approach) --- p.7 / Chapter 1.3.2. --- In vitro inhibition/induction studies --- p.8 / Chapter 1.3.3. --- Clinical studies --- p.9 / Chapter CHAPTER 2. --- Effect of flavonoid-containing herbs on CYP 450 enzyme activities: a screening study in rat --- p.11 / Chapter 2.1 --- Introduction --- p.11 / Chapter 2.2 --- Materials and Methods --- p.12 / Chapter 2.2.1. --- Chemicals --- p.12 / Chapter 2.2.2. --- Herbs --- p.12 / Chapter 2.2.3. --- Preparation of herbal extracts --- p.13 / Chapter 2.2.3.1. --- Preparation of Green Tea extract --- p.13 / Chapter 2.2.3.2. --- Preparation of Decaffeinated Green Tea (DGT) and its extracts --- p.13 / Chapter 2.2.3.3. --- "Preparation of extracts of Huang Qin, Ge Gen and Huai Mi" --- p.14 / Chapter 2.2.3.4. --- Preparation of Ginkgo biloba extract suspension --- p.14 / Chapter 2.2.4. --- Animal treatment --- p.14 / Chapter 2.2.5. --- Preparation of rat liver microsomes --- p.15 / Chapter 2.2.6. --- Determination of protein content of liver microsomes --- p.16 / Chapter 2.2.7. --- Determination of microsomal CYP content --- p.17 / Chapter 2.2.8. --- Statistical analysis --- p.19 / Chapter 2.3 --- Results --- p.19 / Chapter 2.4 --- Discussion --- p.24 / Chapter 2.5 --- Conclusion --- p.25 / Chapter CHAPTER 3 --- Rationale of the clinical study --- p.26 / Chapter CHAPTER 4 --- Development of HPLC methods for simultaneous determination of multiple probe drugs and their metabolites in human plasma or urine --- p.30 / Chapter 4.1 --- Introduction --- p.30 / Chapter 4.2 --- Materials and Methods --- p.33 / Chapter 4.2.1. --- Chemicals and reagents --- p.33 / Chapter 4.2.2. --- Preparation of stock and working solutions --- p.33 / Chapter 4.2.3. --- Equipment and chromatographic conditions --- p.34 / Chapter 4.2.4. --- Treatment of plasma and urine samples with β-glucuronidase --- p.35 / Chapter 4.2.5. --- Extraction procedures --- p.36 / Chapter 4.2.6. --- Preparation of working solutions for calibration curve --- p.37 / Chapter 4.3 --- Results --- p.39 / Chapter 4.3.1. --- Separation of the analytes --- p.39 / Chapter 4.3.2. --- Calibration and linearity --- p.39 / Chapter 4.3.3. --- Sensitivity --- p.39 / Chapter 4.3.4 --- Accuracy and precision --- p.50 / Chapter 4.4 --- Discussion --- p.52 / Chapter 4.5 --- Conclusions --- p.53 / Chapter CHAPTER 5 --- Stability study of probe drugs --- p.54 / Chapter 5.1 --- Introduction --- p.54 / Chapter 5.2 --- Materials and Methods --- p.54 / Chapter 5.2.1. --- Preparation of standard solutions of probe drugs --- p.54 / Chapter 5.2.2. --- Preparation of stability study mediums --- p.54 / Chapter 5.2.2.1. --- Gastric juice (pH=1.2) --- p.54 / Chapter 5.2.2.2. --- Intestine fluid (pH=6.8) --- p.54 / Chapter 5.2.2.3. --- Human plasma (pH=7.4) --- p.55 / Chapter 5.2.2.4. --- Phosphate buffer (pH=7.4) --- p.55 / Chapter 5.2.3. --- Incubation --- p.55 / Chapter 5.2.4. --- Determination of probe drug concentrations in incubation samples --- p.56 / Chapter 5.3 --- Results --- p.57 / Chapter 5.4 --- Discussion --- p.59 / Chapter 5.5 --- Conclusion --- p.59 / Chapter CHAPTER 6 --- Effect of the extract of Ginkgo biloba leaf (761) on CYP isozymes in human subjects --- p.60 / Chapter 6.1 --- Introduction --- p.60 / Chapter 6.2 --- Materials and Methods --- p.60 / Chapter 6.2.1. --- Drugs --- p.60 / Chapter 6.2.2. --- Subjects --- p.61 / Chapter 6.2.3. --- Study design --- p.62 / Chapter 6.2.4. --- Determination of probe drugs/metabolites in the plasma and urine --- p.63 / Chapter 6.2.5. --- Data analysis --- p.65 / Chapter 6.2.6. --- Statistical analysis --- p.66 / Chapter 6.3 --- Results --- p.66 / Chapter 6.3.1. --- Effect of EGb761on CYP1A2 activity --- p.66 / Chapter 6.3.2. --- Effect of EGb761on CYP2E1 activity --- p.67 / Chapter 6.3.3. --- Effect of EGb761 on CYP450 3A activity --- p.68 / Chapter 6.3.4. --- Effect of EGb761 on NAT2 activity --- p.69 / Chapter 6.3.5. --- Effect of EGb761 on CYP2D6 activity --- p.70 / Chapter 6.3.6. --- Effects of EGb761 on CYP2C19 activity --- p.71 / Chapter 6.4 --- Discussion --- p.72 / Chapter 6.5 --- Conclusion --- p.76 / References --- p.77 / Appendix --- p.90

Yim Wan Sze. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 95-109). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 槪論 --- p.iv / Table of contents --- p.v / List of abbreviations --- p.xii / List of Figures --- p.xv / List of Tables --- p.xviii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Oxidation stress --- p.1 / Chapter 1.1.1 --- Free radicals --- p.2 / Chapter 1.1.1.1 --- Hydrogen peroxide --- p.3 / Chapter 1.1.1.2 --- Menadione --- p.4 / Chapter 1.1.2 --- Diseases related to oxidative stress --- p.5 / Chapter 1.1.3 --- Liver Injury --- p.5 / Chapter 1.1.4 --- Antioxidants --- p.7 / Chapter 1.1.4.1 --- Importance of antioxidant --- p.7 / Chapter 1.1.4.2 --- Examples of antioxidant --- p.7 / Chapter 1.2 --- Traditional Chinese Medicinal (TCM) formula Wu-zi-yan-zong-wan (WZ) --- p.8 / Chapter 1.2.1 --- The WZ medicinal formula --- p.8 / Chapter 1.2.2 --- Pharmacological actions of WZ --- p.9 / Chapter 1.2.3 --- Pharmacological actions of individual herbs --- p.10 / Chapter 1.2.3.1 --- Fructus Lycii --- p.10 / Chapter 1.2.3.2 --- Semen Cuscuta --- p.11 / Chapter 1.2.3.3 --- Fructus Rubi --- p.12 / Chapter 1.2.3.4 --- Semen Plantaginis --- p.12 / Chapter 1.2.3.5 --- Fructus Schisandrae --- p.13 / Chapter 1.3 --- The relationship between the liver and kidney --- p.14 / Chapter 1.4 --- Objectives of study --- p.15 / Chapter Chapter 2 --- Preparation of Aqueous Extraction of Wu-zi-yan-zong-wan --- p.17 / Chapter 2.1 --- Introduction --- p.17 / Chapter 2.2 --- Materials and Methods --- p.18 / Chapter 2.2.1 --- Preparation of Wu-zi-yan-zong-wan (WZ) --- p.18 / Chapter 2.2.1.1 --- WZ extracts from raw materials (WZ-e) --- p.18 / Chapter 2.2.1.2 --- WZ extracts from commercial available ready-to-use powders (WZ-p) --- p.20 / Chapter 2.3 --- Results --- p.22 / Chapter 2.3.1 --- Extraction Yield for WZ-e --- p.22 / Chapter Chapter 3 --- In vitro Total Antioxidant Capacity of Aqueous Extracts of Wu-zi-yan-zong-wan and its Components --- p.24 / Chapter 3.1 --- Introduction --- p.24 / Chapter 3.1.1 --- Total Antioxidants: Trolox Equavalent Antioxidant Capacity (TEAC) --- p.24 / Chapter 3.1.2 --- Objectives --- p.25 / Chapter 3.2 --- Materials and methods --- p.26 / Chapter 3.2.1 --- Materials --- p.26 / Chapter 3.2.1.1 --- Reagents --- p.26 / Chapter 3.2.1.2 --- Instruments --- p.26 / Chapter 3.2.2 --- Methods --- p.26 / Chapter 3.2.2.1 --- Total Antioxidnats: Trolox Equavalent Antioxidnat Capacity (TEAC) --- p.26 / Chapter 3.2.3 --- Statistical analysis --- p.27 / Chapter 3.3 --- Results --- p.28 / Chapter 3.3.1 --- Antioxidnat Capacity of Trolox --- p.28 / Chapter 3.3.2 --- Antioxidant capacity of WZ-e formula --- p.28 / Chapter 3.3.3 --- Antioxidant capacity of WZ-p formula --- p.28 / Chapter 3.3.4 --- The total antioxidant capacity of WZ-e and its simplified formulae --- p.32 / Chapter 3.3.5 --- The total antioxidant capacity of WZ-p and its simplified formulae --- p.32 / Chapter 3.3.6 --- Synergetic effect of WZ-e and its simplified formulae --- p.34 / Chapter 3.3.7 --- Orthogonal analysis of WZ-e and its simplified formulae on TEAC assay --- p.40 / Chapter 3.4 --- Discussion --- p.42 / Chapter Chapter 4 --- Antioxidant Activity of Aqueous Extracts of Simplified Formulae of Wu-zi-yan-zong-wan in vitro --- p.44 / Chapter 4.1 --- Introduction --- p.44 / Chapter 4.1.1 --- In vitro antioxidant --- p.44 / Chapter 4.1.2 --- Antioxidant effect of Catechins --- p.44 / Chapter 4.1.3 --- MTT assay --- p.45 / Chapter 4.1.4 --- Objectives --- p.46 / Chapter 4.2 --- Materials and methods --- p.47 / Chapter 4.2.1 --- Materials --- p.47 / Chapter 4.2.1.1 --- Cell Culture --- p.47 / Chapter 4.2.1.2 --- Reagents --- p.47 / Chapter 4.2.2 --- Methods --- p.47 / Chapter 4.2.2.1 --- Cell Culture --- p.47 / Chapter 4.2.2.2 --- MTT Cytotoxicity Assay --- p.48 / Chapter 4.2.3 --- Statistical analysis --- p.48 / Chapter 4.3 --- Results --- p.49 / Chapter 4.3.1 --- The effect of WZ-e formula on HepG2 cells --- p.49 / Chapter 4.3.2 --- The effect of hydrogen peroxide on HepG2 cells --- p.49 / Chapter 4.3.3 --- The effect of menadione on HepG2 cells --- p.52 / Chapter 4.3.4 --- The effect of catechin on HepG2 cells --- p.52 / Chapter 4.3.5 --- The effect of WZ-e and its simplified formulae against hydrogen peroxide-induced cytotoxicty on HepG2 cells --- p.55 / Chapter 4.3.6 --- The effect of WZ-e and its simplified formulae against menadione-induced cytotoxicity on HepG2 cells --- p.60 / Chapter 4.3.7 --- The effect of WZ-p on HepG2 cells --- p.65 / Chapter 4.3.8 --- The effect of WZ-p and its simplified formulae against hydrogen peroxide-induced cytotoxicity on HepG2 cells --- p.67 / Chapter 4.3.9 --- The effect of WZ-p and its simplified formulae against menadione-induced cytotoxicity on HepG2 cells --- p.72 / Chapter 4.4 --- Discussion --- p.77 / Chapter Chapter 5 --- Effect of Aqueous Extract of Wu-zi-yan-zong-wan on Menadione-induced Oxidative Damage in Mouse Liver --- p.79 / Chapter 5.1 --- Introduction --- p.79 / Chapter 5.2 --- Materials and methods --- p.81 / Chapter 5.2.1 --- Materials --- p.81 / Chapter 5.2.1.1 --- Animals --- p.81 / Chapter 5.2.1.2 --- Reagents --- p.81 / Chapter 5.2.1.3 --- Apparatus --- p.81 / Chapter 5.2.2 --- Methods --- p.82 / Chapter 5.2.2.1 --- Animal treatments --- p.82 / Chapter 5.2.2.2 --- Collection of samples --- p.82 / Chapter 5.2.2.3 --- Marker enzyme measurement (ALT) --- p.83 / Chapter 5.2.3 --- Statistical analysis --- p.83 / Chapter 5.3 --- Results --- p.84 / Chapter 5.3.1 --- Dose-dependent effect of WZ-e on menadione hepatotoxicity --- p.84 / Chapter 5.3.2 --- Dose-dependent effect of WZ-e on menadione hepatotoxicity as illustrated by histopathological observations --- p.86 / Chapter 5.3.3 --- Analyzing he effect of WZ-e and its simplified formulae on menadione-induced hepatotoxicity by Orthogonal Analysis89 --- p.89 / Chapter 5.4 --- Discussion --- p.91 / Conclusion --- p.93 / References --- p.95

Made available in DSpace on 2014-06-11T19:22:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-05Bitstream added on 2014-06-13T19:07:30Z : No. of bitstreams: 1 chagas_ha_me_botfca.pdf: 1461839 bytes, checksum: e3cb655599f9ae29650f6b80ac395c35 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / A mamoneira (Ricinus communis L.) é uma espécie oleaginosa tropical, sendo o óleo extraído de suas sementes um dos mais versáteis da natureza e com inúmeras aplicações industriais. Embora ainda seja uma espécie rústica, ela está sujeita a diversas doenças, dentre elas o mofo-cinzento, causada pelo fungo Amphobotrys ricini. O melhoramento genético seria a melhor alternativa para o controle da doença, mas demanda tempo para se obter cultivares resistentes. Dessa Maneira, o uso de métodos de controle baseado em métodos químicos, alternativos ou biológicos mostra-se viável em curto prazo. O objetivo do trabalho foi estudar a eficiência de controle do mofocinzento, na cultura da mamoneira, utilizando-se de métodos químico, alternativo e biológico. Para tanto, foram verificados, in vitro, a eficiência de diferentes meios de cultura na esporulação do patógeno e do antagonista C. rosea. Verificou-se, também, a produção de escleródios do patógeno nos meios. Quanto à eficiência dos métodos, verificou-se, in vitro, a eficiência deles na inibição do crescimento micelial e da germinação dos conídios do patógeno. Após desenvolver e validar uma escala diagramática de avaliação de severidade da doença em racemos de mamoneira, verificou-se a eficiência de aplicação do antagonista C. rosea em frutos destacados da mamona inoculados com o patógeno. Em plantas submetidas a condições de estufa e em campo, verificou-se a eficiência dos métodos no controle da doença causada por A. ricini Quanto à esporulação, o melhor meio de cultura para o patógeno foi V8-20%, obtendo 5,7 x 106 conídios/mL. Para o antagonista C. rosea, o melhor meio foi o TJ- 5% (Suco de Tomate), produzindo 4,41 x 106 conídios/mL. O único meio que propiciou abundante produção de escleródios de A. ricini foi o aveia-ágar. Quanto a inibição do crescimento micelial do patógeno... / The castor bean (Ricinus communis L.) is a tropical oily species, and the oil extracted of its seeds is one of most versatile of the nature, with many industrial applications. Even being a rustic species, it still subjects to several diseases, between them the gray mold, caused by the fungus Amphobotrys ricini. The genetic breeding would be the best alternative for the disease control, but spend time to get a resistant cultivar. In this way, the use of methods of control based on chemical, alternative or biological methods shows viable in short time. The aim of this work was to study the efficiency of the control of gray mold, on castor bean crop, using chemical, alternative and biological methods. Therefore, they had been verified, in vitro, the efficiency of different culture medium in the pathogen sporulation and the antagonist C. rosea. The sclerotia production in the culture medium can be also verified. About the efficiency of the methods, in vitro, the inhibition of the mycelial growth and germination of conidia was verified. After to develop and to validate a diagrammatic scale of evaluation of severity of the disease in racemes of castor bean, the efficiency of application of the antagonist C. rosea in inoculated fruits detached of castor bean with the pathogen was verified. In plants submitted to greenhouse and field conditions, the efficiency of the methods in the control of the disease caused by A. ricini was verified About the sporulation, the optimum culture medium for the pathogen was V8-20%, getting 5,7 x 106 conidia/mL. For the antagonist... (Complete abstract click electronic access below)

Esse trabalho descreve o estudo monitorado de extratos de origem microbiológica e vegetal. Com o objetivo de identificar compostos com atividade antioxidante e/ou inibidores da enzima acetilcolinesterase em extratos de origem microbiológica e vegetal do cerrado brasileiro, uma triagem de atividade foi realizada utilizando ensaios simples e rápidos. Nessa triagem dois extratos promissores foram selecionados para os estudos de identificação dos compostos responsáveis pela atividade inicial. O trabalho de purificação foi iniciado com a fração em acetato de etila do extrato etanólico da actinobactéria-36 (50PL), Streptomyces sp., fermentado em meio de canjica amarela que apresentou atividade nos dois ensaios realizados. As atividades antioxidante e anticolinesterásica são relatadas pela primeira vez para essa actinobactéria. Nesse estudo foram identificados dois compostos, o éster metílico do ácido cis-6, cis-8 octadecadienóico e o tetradecanal. Da espécie Psychotria carthagenensis, uma planta da família Rubiaceae, foram objeto de estudo as frações hexânica e acetato de etila oriundas do extrato etanólico das folhas, o extrato hexânico das folhas e o extrato etanólico dos caules. A espécie P. cartahgenensis foi investigada quanto à presença de alcalóides uma vez que é utilizada juntamente com as espécies Psychotria viridis e Banisteriopsis caapi no preparo de uma bebida alucinógena conhecida como ayahuasca. A partir dos extratos etanólicos das sementes, caules e folhas foi realizada uma extração ácido-base resultando em frações ricas em compostos nitrogenados. As frações de alcalóides totais foram analisadas em TLC e revelador específico, o cloro-iodoplatinado, evidenciando a presença de alcalóides. As frações foram analisadas por EM (desreplicação) resultando na identificação de 5 compostos nitrogenados. / This work describes the monitored study of extracts from microbiological and plant origin. In order to identify compounds with antioxidant action and/or inhibitors of acetylcholinesterase enzyme in extracts of microbial and plants of the Brazilian Cerrado vegetation, screening for these activities was performed using simple and rapid tests. From this screening, two promising extracts were selected for identification of the compounds responsible for the initially observed activity. Purification was started with the ethyl acetate fraction in the ethanol extract of actinobacteria-36 (50PL), Streptomyces sp., fermented in a yellow hominy culture medium that displayed activity in both tests. Antioxidant and anticholinesterase activities are reported for this actinobacteria for the first time. Two compounds were identified, namely 6(Z),8(Z)-octadecadienoic acid, methyl ester and tetradecanal. The hexane and ethyl acetate fractions of the ethanol extract of the leaves as well as the ethanol extract of the stems from the Psychotria carthagenensis species, a plant of the Rubiaceae family, were studied. This species was investigated for the presence of alkaloids, because it is used together with the species Psychotria viridis and Banisteriopsis caapi in the preparation of a hallucinogenic drink known as ayahuasca. Acid-base extractions of the ethanol extracts of the seeds, stems, and leaves of this plant were carried out, resulting in fractions rich in nitrogen compounds. The total alkaloids fractions were analyzed by TLC and specific revealing with chlorine-iodoplatinate, which evidenced the presence of alkaloids. The fractions were analyzed by MS (derreplication), which allowed for identification of five nitrogen compounds.

Made available in DSpace on 2017-07-10T17:37:29Z (GMT). No. of bitstreams: 1 Mauricele Baldo.pdf: 791599 bytes, checksum: bae07fffa63ea2af6fe2884dfd9c37e6 (MD5) Previous issue date: 2008-03-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / One of the focus of the alternative agriculture is the disease control, wich include biological control, the induction of resistance in plants and the use of the natural products with antimicrobial activity and /or resistance induction. Basidiocarps extracts have been studied for disease control. This research focused the avaliation of the fungitoxic effect in the aqueous extracts of the mycelium (EAM), basidiocarps (EAB) and filtrate of culture (EAF) of Pycnoporus sanguineus, in the Colletotrichum lindemuthianum, and avaliate the control of anthracnose in the bean plant by theses extracts and the resistance induction through determination of activity of the enzyme peroxidase (POX), polyphenol oxidase (PFO), phenylalanine ammonia-lyase (PAL) and β-1,3 glucanase, besydes detection in situ of the reatives oxygen species (ROS) H2O2 and O2-. For the tests in vitro were utilized the extracts on concentration 1, 5, 10, 15 and 20%, avaliating the micelial growth and the germination of the C. lindemuthianum spores. In order to severity tests and determination of enzyme, extracts on concentrations 5 and 10% were applayed on the 1st trifoliate leaves of bean plant, three days before of the inoculation of the pathogen (1x104 conidia mL-1) on 1st s and 2nd s leaves. The same procediment was adopt for detection of ROS, instead utilized just EAM 5%, water, acibenzolar-S-methyl (150 mg i.a.L-1) and azoxystrobin fungicide (4 g. i. a. L-1). For the enzyme determination was amostrated the 1st s and 2nd s leaves at the moment of treatment and after 3, 6, 9, 12, 27 days. For ROS was made avaliations to 48, 96 and 192 hours after the inoculation. The results indicated direct antimicrobial activity of the extracts, having these extracts inhibited the germination, but they didn t inhibited the micelial growing. On plants, the EAF 5 e 10%, reduced 69 and 67% respectively, the severity on the 1st s leaves, while the fungicide and ASM reduced 87 and 76% respectively. On 2nd s leaves the more significative reductions were observed to EAB 10% and EAF 5 and 10%, with reduction of the severity of 63%. This reduction on severity can be associated with POX, PAL and β-1,3 glucanase what apresented alterations on the especific activity, what wasn t observed to PFO. The formation of H2O2 wasn t detected, on 48 and 96 hours. In 192 hours the EAB treatment apresented citoplasmatic desorganization and collapse of epidermic cell similar to the water control. In times 48 and 96 hours there was of the O2- in the EAM, EAB and ASM treatment. In 192 all of the treatments apresented reaction to O2- in epidermic cells and the mesophyll. This results apoint the potencial of P. sanguineus extracts for the control of in the anthracnose in bean plants, that can occurr by induction of the local and systemic resistance an/or antimicrobial direct activity / Um dos enfoques da agricultura alternativa é o controle alternativo de doenças, o qual inclui o controle biológico, a indução de resistência em plantas e o uso de produtos naturais com atividade antimicrobiana e/ou indutora de resistência. Extratos de basidiocarpos têm sido estudados no controle de doenças. Este trabalho teve como objetivos avaliar o efeito fungitóxico dos extratos aquosos de micélio (EAM), basidiocarpo (EAB) e filtrado de cultura (EAF) de Pycnoporus sanguineus, sobre Colletotrichum lindemuthianum; e avaliar o controle da antracnose em feijoeiro por estes extratos, bem como a indução de resistência pela determinação da atividade das enzimas peroxidase (POX), polifenoloxidase (PFO), fenilalanina amônia-liase (FAL) e β-1,3 glucanase; além de detecção in situ das espécies reativas de oxigênio (ERO s) H2O2 e O2-. Para os ensaios in vitro foram utilizados os extratos nas concentrações 1, 5, 10, 15 e 20%, avaliando-se o crescimento micelial e germinação de esporos de C. lindemuthianum. Para os ensaios de severidade e determinação das enzimas, extratos nas concentrações 5 e 10% foram aplicados nas 1as folhas trifoliadas de feijoeiro, três dias antes da inoculação do patógeno (1x104 conídios mL-1) realizada nas 1as e 2as folhas. O mesmo procedimento foi adotado para detecção das ERO s, porém utilizados apenas EAM e EAB 5%. Água, acibenzolar-S-metil (75 mg i. a. L-1) e fungicida azoxystrobin (4 g i. a. L-1) foram utilizados como tratamentos controle para todos os ensaios. Para a dosagem das enzimas foram amostradas as 1as e 2as folhas no momento dos tratamentos e após 3, 6, 9, 12 e 27 dias. Para ERO s foram feitas avaliações às 48, 96 e 192 h após inoculação. Os resultados indicaram atividade antimicrobiana direta dos extratos, tendo estes inibido a germinação, porém não apresentaram efeito inibitório do crescimento micelial. Nas plantas, os EAF 5 e 10%, reduziram 69 e 67%, respectivamente, a severidade nas 1as folhas, enquanto o fungicida e ASM reduziram 87 e 76%, respectivamente. Nas 2as folhas as maiores reduções foram observadas para EAB 10% e EAF 5 e 10%, com redução média da severidade de 63%. Esta redução na severidade pode estar associada com POX, FAL e β-1,3 glucanase que apresentaram alterações na atividade específica, o que não foi observado para PFO. Não foi detectada formação de H2O2 nas 48 e 96 h. Em 192 h os tratamentos EAB e EAM apresentaram desorganização citoplasmática e colapso das células epidérmicas semelhantes ao controle água. Nos tempos 48 e 96 h houve formação de O2- nos tratamentos EAM, EAB e ASM. Em 192 h todos os tratamentos apresentaram reação para O2- nas células epidérmicas e do mesófilo. Estes resultados indicam o potencial dos extratos de P. sanguineus no controle da antracnose do feijoeiro, que pode ocorrer tanto por indução de resistência local e sistêmica e/ou atividade antimicrobiana direta

Made available in DSpace on 2017-07-10T17:37:48Z (GMT). No. of bitstreams: 1 Ana_Paula_Mamprim.PDF: 1025073 bytes, checksum: a4585bdee440fbcff474cdb4a0e84089 (MD5) Previous issue date: 2011-08-25 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objective of this study was to evaluate the compatibility of aqueous and alcoholic plant extracts, alternative products and basidiocarps of the fungus Pycnoporus sanguineus on Metarhizium anisopliae. These plants and the basidiocarps were collected and transferred to drying oven at 40°C for 7 days and then ground into a fine powder. The extracts and basidiocarps were used in 10% concentration and the alternative products in 3 concentrations, being established on product labels (CR), half (0,5CR) and twice (2CR). All treatments were sprayed on the already inoculated fungus in PDA culture medium. Viability was evaluated by direct counting of viable and unviable conidia after incubation for 16 h at 26 ± 1 º C, 12h photoperiod. For Colony Forming Unit (CFU), it was counted the number of colonies after 5 days of incubation. To vegetative growth and production of conidia, the fungus was inoculated in three points on the culture medium surface of each petri dish, staying for 7 days for subsequent colonies measurement and counting conidia. For the viability was found that all aqueous extracts, except the chinaberry differed from the control. For the CFU, only cinnamon and laurel extracts and P.sanguineus not differed from the control. In diameter, lemon grass, rue, chinaberry, cinnamon, citronella and rosemary extracts differed from the control. For conidiogeneous, lemon grass, rue, cinnamon and rosemary extracts and P. sanguineus also had significance in relation to control. All the alcoholic extracts, for viability, differed from the control. However, for CFU these same extracts showed no significance. For the diameter, it was a significant difference to the alcoholic extracts of rue, castor, neem and chinaberry. For the conidia production there were significant difference between alcoholic extracts of rue, castor, chinaberry, cinnamon, neem, rosemary and laurel. For the alternative products, in the viability parameter, all products showed significant difference in relation to control, except Forth (0,5 CR and CR) and Agro-Mos® in CR. For CFU, there was significant difference at all concentrations tested. For the diameter, only 0,5CR Pironim® differed at the control. For the conidia production, Plant Clean® (0.5 CR and CR), Pironim® (CR), Bordeaux Mixture (0.5 CR and CR) and Sulfur Mixture (CR and 2CR) showed significant differences compared to control. Despite of the biological parameters present percentage change (reduction or stimulation), there was compatibility for aqueous and alcoholic extracts. In relation to alternative products, only Sulfur Mixture was incompatible for the fungus M. anisopliae. / O objetivo deste trabalho foi avaliar a compatibilidade dos extratos vegetais aquosos e alcoólicos e de produtos alternativos, além de extratos de basidiocarpos de Pycnoporus sanguineus sobre o fungo entomopatogênico Metarhizium anisopliae. As plantas e os basidiocarpos foram coletados e transferidos para estufa de secagem em 40 ºC por 7 dias, sendo em seguida, moídos até se obter um pó fino. Os extratos e o basidiocarpos foram utilizados na concentração 10%, e os produtos alternativos em três concentrações, sendo a estabelecida nos rótulos dos produtos (CR), a metade (0,5CR) e o dobro da mesma (2CR). Em todos os tratamentos foram feitas pulverizações sobre o fungo já repicado no meio de cultura BDA. Avaliou-se viabilidade, por meio da contagem direta dos conídios viáveis e inviáveis após incubação por 16 h em 26±1ºC, fotofase 12h. Para Unidade Formadora de Colônias (UFC) contou-se o número de colônias após 5 dias de incubação. Em relação ao crescimento vegetativo e a produção de conídios, o fungo foi repicado em 3 pontos na superfície do meio de cultura de cada placa de Petri, permanecendo em incubação por 7 dias, para posterior medição das colônias e contagem de conídios. Verificou-se que todos os extratos aquosos, exceto o de cinamomo, diferiram da testemunha para viabilidade. Para o UFC, somente os extratos de canela, louro e P.sanguineus, não diferiram da testemunha. No diâmetro, os extratos de capim cidreira, arruda, cinamomo, canela, citronela, alecrim diferiram da testemunha. Assim como na conidiogênese onde os extratos capim cidreira, arruda, canela, alecrim e P sanguineus também apresentaram significância em relação à testemunha. Para os extratos alcoólicos observou-se que na viabilidade, todos diferiram da testemunha. Enquanto que na UFC esses mesmos extratos não apresentaram significância. Para o diâmetro ocorreu uma diferença significativa para os extratos alcoólicos de arruda, mamona, cinamomo e nim. Já para a produção de conídios verificou-se diferença significativa dos extratos alcoólicos arruda, mamona, cinamomo, canela, nim, alecrim e louro. Para os produtos fitossanitários alternativos, no parâmetro viabilidade, todos os produtos apresentaram diferença significativa em relação a testemunha, exceto Forth (0,5 CR e na CR) e Agro-mos® na CR. Na UFC, foi verificada diferença significativa em todas as concentrações testadas. Para o diâmetro apenas o Pironim® na 0,5CR diferiu da testemunha. Já para a produção de conídios o produto Planta Clean® (0,5 CR e na CR), Pironim® (CR), Calda Bordalesa (0,5 CR e na CR) e Calda Sulfocálcica (CR e no 2CR) apresentaram diferenças significativas dos tratamentos em relação à testemunha. Apesar dos parâmetros biológicos apresentarem variação percentual (estímulo ou redução) verificou-se a compatibilidade para os extratos aquosos e alcoólicos. Em relação aos produtos alternativos somente a Calda Sulfocálcica se mostrou incompatível para o fungo M. anisopliae.