Immortalized normal hepatocytes AML-12 or primary hepatocytes were cultured in control, and the methionine and choline deficient (MCD) culture medium in the presence or absence of Phyllanthus urinaria for 24 hours. Hepatocyte triglyceride contents, release of alanine aminotransferase, lipoperoxides and reactive oxygen species production were determined in the cell culture study. Age-matched wild-type C57BL/6 and diabetes db/db mice were fed control or MCD diet for 10 days with or without Phyllanthus urinaria. The levels of Hepatic steatosis, necroinflammation, triglycerides and oxidative stress were investigated. Hepatic expression of inflammatory factors and lipid regulatory mediators were assayed. The results demonstrated that Phyllanthus urinaria reduced steatosis and alanine aminotransferase (ALT) levels in culture of hepatocytes in a dose-dependent manner. Phyllanthus urinaria protected the livers against MCD-induced hepatic fat accumulation and steatohepatitis in mice. This effect was associated with repressed levels of hepatic lipid peroxides, reduced expression of cytochrome P450 (CYP) 2e1, pro-inflammatory tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), dampened activation of inflammatory C-jun N-teuninal kinase (JNK) and nuclear factor kappaB (NF-kappaB), increased expression of lipolytic Cyp4a10 and suppressed transcriptional activity of lipogenic CCAAT/enhancer binding protein beta (C/EBPbeta). Hepatic acyl co-enzyme A oxidase (ACO) that regulated hepatic beta-oxidation of fatty acid and other lipid regulators were not affected by Phyllanthus urinaria. / Non-alcoholic steatohepatitis (NASH) results from excessive accumulation of hepatic fat (steatosis) and oxidative stress. Therefore, inhibition of fatty acid cytotoxicity and liver inflammtary change is an important goal in the treatment of NASH. Phyllanthus urinaria, a herbal medicine, has been reported to have potential anti-oxidant property. We tested the effects of Phyllanthus urinaria on nutritional steatohepatitis both in vitro and in vivo, and determined the mechanism of its action. / Our study indicated that Phyllanthus urinaria effectively prevented MCD-induced steatohepatitis. This effect were probably mediated through dampening oxidative stress, ameliorating inflammation and decreasing lipid accumulation. Phyllanthus urinaria deserves further evaluation for its potential therapeutic effect on NASH in humans. / Shen, Bo. / Adviser: Henry Ly Chan. / Source: Dissertation Abstracts International, Volume: 70-09, Section: B, page: . / Thesis submitted in: November 2008. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 128-142). / 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.

Ganoderma lucidum and Cordyceps sinensis are medicinal fungi commonly used in Chinese medicine. The allied species of G. lucidum, G. sinense and G. tsugae as well as the allied species of C. sinensis, C. militaris are also commercially available as health supplements. The present study aimed at evaluating and comparing the biological activities of the different parts of fruiting bodies of Ganoderma species (whole fruiting body, pileus, stipe, spores and spore oil) and Cordyceps species (whole fruiting body, stroma and larva/pupa). / The extracts of C. sinensis and C. mililaris could stimulate secretion in human airway epithelial cell Calu-3, implying the potentials of these extracts to modulate the mucociliary clearance. The potencies of the stimulatory effects of the stroma of fruiting bodies showed stronger effects than the larva/pupa. All parts of C. sinensis and the stroma of C. mililaris could modulate the proliferation of human peripheral blood mononuclear cells and the different potencies of immunomodulatory effects were also observed. / The results demonstrated that the hot water extracts of G. lucidum, G. sinense and G. tsugae possessed antiproliferative effects on human breast cancer cell lines MCF-7 and MDA-MB-231. The extracts from the stipes showed stronger inhibitory activities on cancer cell proliferation than those from pilei. Furthermore, the extracts from the whole fruiting body and stipe of G. lucidum possessed strong antitumor effects on the nude mice bearing MCF-7 xenografts and the BALB/c mice bearing sarcoma S-180 allografts as well as strong immunomodulatory effects in terms of stimulating splenic lymphocyte proliferative responses. The oral administrations of spores and spore oil did not show significant inhibition on MCF-7 xenografts growth but they inhibited sarcoma allografts growth effectively. / The strong biological effects of the stipe of Ganoderma and the stroma of Cordyceps were showed for the first-time. This study sheds light on how the fruiting bodies of Ganoderma or Cordyceps should be used to achieve the most out of their pharmacological properties. This study also demonstrated the novel application of Cordyceps in promoting secretion in human airway submucosal glands, which reinforces the rationale of using this fungus for treating respiratory diseases. / Yue Gar Lee Grace. / "August 2006." / Advisers: Leung Ping Chung; Kwok Pui Fung; Bik San Clara Lau. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1584. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 318-340). / 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.

As previsões mais otimistas para o mundo no segundo milênio alertam para a escassez de água. Esta terá como origem a redução de volume de água doce aproveitável, o desperdício e poluição das fontes de água. O cloro, usado para a desinfecção, se combina com os resíduos de matéria orgânica remanescente do tratamento da água de consumo formando os organoclorados (produtos potencialmente cancerígenos). A proposta deste trabalho, é oferecer subsídio à sanitização da água buscando, pelo resgate etnográfico, recursos naturais sustentáveis, renováveis e ecologicamente corretos como alternativa para o tratamento da água na pequena propriedade rural. Das dezoito plantas indicadas e triadas na forma de extratos hidro-alcoólicos quanto a Intensidade de Atividade Antibacteriana (IAAB), selecionou-se os cinco que tiveram o melhor desempenho no controle das bactérias em teste (duas gram positivas e duas gram negativas). Os extratos escolhidos passaram pelo teste de Concentração Inibitória Mínima/Concentração Bactericida Mínima (CIM/CBM). O que mais se destacou foi o extrato de Sete Sangrias (Cuphea Carthagenensis (Jacq.) Macbride) que controlou as bactérias desafiantes até a concentração de 10%. Estes extratos também foram testados quanto à citotoxicidade e constatou-se que a Baleeira (Cordia curassavica), Folha da Fortuna (Bryophyllum pinatum Kurz.) e a Sete Sangrias (Cuphea cathagenensis (Jacq.) Macbride) foram citotóxicas em concentrações entre 1 e 10 % sem diferença estatística para o teste de Duncam (p=0,99).Embora a confrontação da capacidade desinfetante e o efeito tóxico apresentados pelos extratos de plantas sejam considerados animadores, não os recomendamos para uso como desinfetantes de volumes hídricos uma vez que a concentração mais baixa efetiva foi a de 10%. / The most optimistic forecast for the world in the second millennium calls attention to the water shortage. This shortage will have as its origin the reduction of volume of useable fresh water, waste and pollution of the water resources. The chlorine, used for disinfection, combines with the residues of organic matter remaining from the treatment of water for consumption forming organochlorides (potentially carcinogenic products). The aim of this work is to offer options to the sanitation of water searching, by ethnographic rescue, natural resources which are sustainable, renewable and ecologically correct as an alternative for water treatment in small rural farms. From the eighteen plants indicated and screened in the form of hydroalcoholic extracts regarding their Antibacterial Activity Intensity (IAAB), the five that showed better performance in the control of bacteria (two Gram positive and two Gram negative), were selected. The chosen extracts were submitted to the test of Minimal Inhibitory Concentration/ Minimal Bactericide Concentration (CIM/CBM). The one that showed the best performance was Sete Sangrias (Cuphea Carthagenensis (Jacq.) Macbride), which controlled the tested bacteria dilution up to 10%. These extracts were also tested regarding their cytotoxicity and it was verified that Baleeira (Cordia curassavica), Folha da Fortuna (Bryophyllum pinatum Kurz.) and Sete sangrias (Cuphea cathagenensis (Jacq.) Macbride) were cytotoxic in concentrations between 1 and 10%, without statistic difference by the Duncam test (p=0,99). Although the confrontation of disinfection capacity and the toxic effect presented by the plant extracts are considered encouraging, we do not recommend them for use as disinfectants of water, as the lowest effective concentration obtained was 10%.

Submitted by Isabela Amendola (isabelaamendola@hotmail.com) on 2019-02-01T11:22:14Z No. of bitstreams: 1 Tese-Isabela-Amêndola.pdf: 2013510 bytes, checksum: 69df8d92f153155f71d6e8a4b37d5eb6 (MD5) / Approved for entry into archive by Silvana Alvarez null (silvana@ict.unesp.br) on 2019-02-01T17:50:14Z (GMT) No. of bitstreams: 1 amendola_i_dr_sjc.pdf: 2013510 bytes, checksum: 69df8d92f153155f71d6e8a4b37d5eb6 (MD5) / Made available in DSpace on 2019-02-01T17:50:14Z (GMT). No. of bitstreams: 1 amendola_i_dr_sjc.pdf: 2013510 bytes, checksum: 69df8d92f153155f71d6e8a4b37d5eb6 (MD5) Previous issue date: 2018-12-03 / A resistência microbiana aos antibióticos disponíveis é preocupação constante, devido à dificuldade no tratamento de infecções causadas por cepas resistentes, em decorrência do uso indiscriminado de antimicrobianos. Assim, a busca por terapias antimicrobianas alternativas tem sido crescente e necessária, sendo a fitoterapia umas das opções de escolha. O objetivo do presente estudo foi analisar a atividade antibacteriana de extratos glicólicos de Achyrocline satureioides (macela), Cynara scolymus (alcachofra), Hamamelis virginiana (hamamelis) e Persea americana (abacateiro), pelos períodos de 5 min e 24 h de exposição sobre bactérias anaeróbias Fusobacterium nucleatum subsp. nucleatum, Parvimonas micra, Porphyromonas endodontalis e Porphyromonas gingivalis, em culturas planctônica e biofilmes. As bactérias armazenadas a -80ºC foram ativadas em caldo Brucella enriquecido (hemina 1%, menadiona 1% e sangue de carneiro desfibrinado 5%) e incubadas em câmara de anaerobiose por 48 h a 37ºC por sete dias. A partir de culturas puras, o teste de microdiluição em caldo foi conduzido em microplacas por meio de suspensões bacterianas padronizadas em solução fisiológica estéril (NaCl 0,9%) e diluições dos extratos em caldo, sendo as placas incubadas por 48 h a 37ºC em anaerobiose. Alíquotas de cada poço foram semeadas em ágar Brucella enriquecido. Após incubação, a Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM) foram determinadas. As concentrações efetivas de cada extrato foram aplicadas sobre os biofilmes de cada espécie, formados em microplacas a partir de suspensões bacterianas puras padronizadas na escala 0,5 de McFarland. As microplacas foram incubadas por sete dias a 37ºC para formação dos biofilmes, sendo o meio trocado a cada 48 h. Os biofilmes foram tratados por 5 min e 24 h. Em seguida, foram lavados e desprendidos por homogeneizador ultrassônico. As suspensões diluídas foram adicionadas em ágar Brucella enriquecido. Após 48 h, as Unidades Formadoras de Colônia por mililitro (UFC/mL) foram determinadas. Os resultados foram analisados por ANOVA e teste de Tukey ou por Kruskal-Wallis e teste Dunns, ambos com nível de significância de 5% (p≤0,05). Sobre as culturas planctônicas, a CIM e CBM dos extratos foi determinada apenas para F. nucleatum. A CBM dos extratos de A. satureioides, C. scolymus e P. americana foi obtida sobre P. micra. Não foi obtida atividade bactericida para P. endodontalis e P. gingivalis. Sobre biofilmes, todas as espécies apresentaram reduções significativas quando expostas aos extratos em ambos os tempos. Pode-se concluir que os extratos testados apresentaram efeito bacteriostático sobre F. nucleatum. Atividade bactericida dos extratos foi observada sobre F. nucleatum, bem como sobre P. micra, exceto para H. virginiana. Os extratos avaliados também apresentaram efeito antibiofilme sobre F. nucleatum, P. micra, P. endodontalis e P. gingivalis por 5 min e 24 h de exposição. / Microbial resistance to antibiotics available is constant concern, due to the difficulty in treating infections caused by resistant strains as a result of the indiscriminate use of antimicrobials. Thus, the search for antimicrobial alternative therapies has been growing and necessary, being one option the herbal medicine. The objective of the present study was to analyze the antibacterial activity to Achyrocline satureioides glycolic extracts (macela), Cynara scolymus (artichoke), Hamamelis virginiana (Witch-Hazel) and Persea americana (avocado), for periods of 5 min and 24 h from exhibition on anaerobic bacteria Fusobacterium nucleatum subsp. nucleatum, Parvimonas micra, Porphyromonas gingivalis and Porphyromonas endodontalis in planktonic communities and biofilms. Bacteria stored at -80°C have been activated in Brucella broth enriched (hemin 1%, menadione 1% and defibrinated sheep blood 5%) and incubated in anaerobiose chamber for 48 h at 37° C for seven days. From pure cultures, the microdiluição test in broth was conducted in microplates through standardized bacterial suspensions in sterile saline solution (NaCl 0.9%) and dilution of the extracts in broth, being incubated plates for 48 h at 37° C in anaerobiosis. Aliquots of each well were sown in Brucella agar enriched. After incubation, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined. Effective concentrations of each extract were applied on the biofilms of each species, formed in microplates from pure bacterial suspensions in 0.5 McFarland scale standard. The microplates were incubated for 7 days at 37°C for the formation of biofilms, being the culture medium replaced every 48 h. Biofilms were treated for 5 min and 24 h have been washed and given off by ultrasonic homogenizer. Dilute suspensions were added in Brucella agar enriched. After 48 h, the Colony Forming Units per milliliter (CFU/ml) were determined. The results were analyzed by ANOVA and Tukey test, or Kruskal-Wallis test and Dunns, both with a significance level of 5% (p ≤ 0.05). On the planktonic cultures, CIM and CBM of extracts was determined only to F. nucleatum. The CBM of the extracts of A. satureioides, C. scolymus and P. americana was obtained on P. micra. Bactericidal activity was not obtained for P. endodontalis and P. gingivalis. About biofilms, all species exhibited significant reductions when exposed to the extracts in both times. It can be concluded that the extracts tested showed bacteriostatic effect on F. nucleatum. Bactericidal activity of extracts was observed on F. nucleatum and P. micra, except for H. virginiana. The extracts evaluated also presented antibiofilme effect on F. nucleatum, P. micra, P. endodontalis and P. gingivalis for 5 min and 24 h.

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

A raiz pulverizada de Pfaffia paniculata e seu extrato butanólico apresentam propriedades antineoplásicas, quimiopreventivas e antiangiogênicas, onde muitos indícios conferem às saponinas triterpenóides presentes nestas raízes as propriedades antitumorais observadas. Sabe-se que saponinas de diversos tipos de plantas possuem capacidade de interferir diretamente no ciclo celular de células tumorais. Assim, considerando os efeitos inibitórios das raízes e extratos de P. paniculata sobre a proliferação celular, o objetivo deste trabalho foi compreender os mecanismos envolvidos na ação quimiopreventiva desta raiz, tanto na fase de iniciação da carcinogênese hepática, quanto sobre células tumorais já estabelecidas. Inicialmente, foram avaliados os efeitos de diferentes concentrações da raiz em pó de P. paniculata adicionada à ração em camundongos submetidos ao modelo de hepatocarcinogênese, pela análise da proliferação celular, indução da apoptose e a comunicação intercelular hepática. Seqüencialmente, foram avaliados os efeitos das frações purificadas do extrato butanólico destas raízes sobre linhagem de células de carcinoma hepatocelular humano. Neste experimento foram analisadas a influência do tratamento sobre a viabilidade celular, fases e proteínas do ciclo celular, proliferação e presença de morte celular. No modelo de hepatocarcinogênese o tratamento com a raiz reduziu a proliferação celular, aumentou a apoptose e desencadeou processo inflamatório crônico em intensidades dependentes da concentração testada e não afetou a comunicação intercelular. Estes resultados indicam que os efeitos quimiopreventivos da P. Paniculata são decorrentes do controle da proliferação celular e apoptose, e são diretamente influenciados pela concentração da raiz. No experimento in vitro o tratamento reduziu a concentração das células vivas sem aumentar a concentração de células mortas, diminuiu a porcentagem de células na fase G2 do ciclo celular, reduziu a expressão dos genes das proteínas ciclinas D1, E e CDK6 e aumentou a expressão de p27, e não induziu apoptose. Estes resultados mostraram que a redução na concentração de células observadas após o tratamento com a fração do extrato butanólico de P. paniculata não foi decorrente de indução de apoptose. O tratamento parou o ciclo celular das células tumorais HepG2 em G1, inibindo proteínas importantes para a progressão do ciclo e estimulando a expressão de p27 um conhecido gene inibidor de CDKs. Os efeitos antiproliferativos observados no experimento in vivo em camundongos, reproduziram-se in vitro em células tumorais humanas, o que pode indicar que as propriedades antineoplásicas anteriormente observadas não são espécie específica. Em linhas gerais, as raízes e/ou as frações do extrato butanólico obtido a partir das raízes de P. paniculata apresentam propriedades antineoplásicas por inibir a proliferação celular e induzir apoptose in vivo e por parar o ciclo celular in vitro. Estes resultados consagram as propriedades antineoplásicas de Pfaffia paniculata e motivam o desenvolvimento de mais estudos sobre as suas potencialidades. / The powdered roots of Pfaffia paniculata and their butanolic extract present antineoplastic, chemopreventive and antiangiogenic properties, and many evidences suggest that the triterpenoid saponins are the responsible for these properties. It is well known that saponins from several types of plants have the capacity to directly interfere on the tumor cell cycle. Therefore, considering the inhibitory effects of the roots and extracts of P. paniculata on cell proliferation, the aim of this study was to search for the mechanisms involved in the chemopreventive effects of this root, both in the initiation phase of the hepatocarcinogenesis and on tumor cell lineage. Initially, the effects of different concentrations of the powdered root of P. paniculata added to the mouse food were evaluated in mice submitted to hepatocarcinogenesis model. Cell proliferation, induction of apoptosis, and hepatic intercellular communication were evaluated. The effects of the purified fractions of the butanolic extract of these roots were then evaluated in human hepatocarcinoma cells. In this experiment, the influence of the treatment on cell viability, phases and proteins of cell cycle, cell proliferation and cell death were evaluated. In the hepatocarcinogenesis model, the treatment with the root decreased cell proliferation, increased apoptosis and induced a chronic inflammatory process dependent on the concentration tested, and did not affect cell communication. These results indicate that the chemopreventive effects of P. Paniculata are apparently dependent on the control of cell proliferation and apoptosis and are directly influenced by the concentration of the root. In the in vitro treatment, it has been observed a reduction in the concentration of live cells without increasing the concentration of dead cells, decreased the level of G2 phase cells, reduced the expression of proteins cyclins D1, E and CDK6, increased the expression of p27, and did not induce apoptosis. These results showed that the reduction in the concentration of cells after the treatment with the butanolic extract of P. paniculata was not due to induction of apoptosis. The treatment inhibited the progression of the cell cycle of HepG2 cells in phase G1, by the inhibition of the expression of proteins that are important to the progression of the cycle, and stimulating the expression of p27, a known inhibitor of CDKs. The antiproliferative effects observed in the in vivo experiments were repeated in the in vitro study with human tumor cells. This may indicate that the antineoplastic properties previously observed are not species-specific. In conclusion, the roots and/or butanolic extract obtained from P. paniculata present antineoplastic properties due to inhibition of cell proliferation and induction of apoptosis in vivo, and due to the cell cycle arrest in vitro. These results reinforce the antineoplastic properties of Pfaffia paniculata and motivate the development of more studies focusing on its antineoplastic potentials.

Submitted by Aline Batista (alinehb.ufpel@gmail.com) on 2018-04-13T18:31:46Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Aline_Silveira_Cardoso_Oliveira.pdf: 2264789 bytes, checksum: 935fe6affb42c7bd8ef6d0e2d3a89651 (MD5) / Made available in DSpace on 2018-04-13T19:59:09Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Aline_Silveira_Cardoso_Oliveira.pdf: 2264789 bytes, checksum: 935fe6affb42c7bd8ef6d0e2d3a89651 (MD5) Previous issue date: 2006-09-12 / Sem bolsa / A prática da fitoterapia segura não se verifica apenas por meio da análise do produto final, mas também, na obtenção da espécie vegetal, desde sua identificação, cultivo, colheita, beneficiamento, armazenamento e comercialização. A ausência de qualidade, a adulteração e a utilização incorreta podem interferir na eficácia e até mesmo na segurança do uso do produto. Uma das espécies amplamente utilizada tanto na medicina popular quanto no sistema oficial de saúde é a Maytenus ilicifolia Mart ex Reissek (espinheira-santa) para o tratamento de dispepsias. O objetivo deste trabalho foi avaliar os parâmetros de qualidade das amostras secas de espinheira-santa disponíveis nos locais de comercialização formal na cidade de Pelotas. Este estudo tem delineamento classificado como descritivo experimental e analítico. Foram coletadas 11 amostras de plantas medicinais popularmente conhecidas como cancorosa e/ou espinheira-santa, vendidas no comércio formal (farmácias, drogarias e supermercados) e uma amostra padrão coletada no Instituto Federal Sul-rio-grandense, Campus Pelotas - Visconde da Graça. Foram avaliadas características organolépticas e físico-químicas, além dos rótulos dos produtos. Todas as amostras foram reprovadas em pelo menos dois parâmetros analisados, indicando que é necessário ampliar a fiscalização visando garantir a segurança ao consumidor. / The practice of safe phytotherapy does not occur only through the analysis of the final product, but also in obtaining the plant species, from its identification, growing, harvest, processing, storage, and commercialization. The lack of quality, adulteration, and the misuse may interfere with the efficacy and even in the safe use of the product. One of the widely used species is Maytenus ilicifolia Mart ex Reissek (espinheira-santa), which is a common practice both in popular medicine and official health system for treatment of dispepsy. The objective of this work was evaluate the parameters of quality of dry samples of „espinheira-santa‟ available in formal local marketing in the municipality of Pelotas. The design of this study is descriptive, experimental and analytical. Eleven samples of medicinal plants popularly known as „cancorosa‟ and/or „espinheira-santa‟ were collected from formal marketing (pharmacies, drugstores and super markets), and a standard sample was collected from Instituto Federal Sul Riograndense, Campus Pelotas – Visconde da Graça. Physico-chemical and organoleptic characteristics were evaluated, as well as the labels of the products. All samples were not approved at least in two parameters analyzed, indicating that is necessary to expand surveillance to ensure consumer safety.

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Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2016-03-15T13:24:29Z No. of bitstreams: 1 PDF - Eveline Angélica Lira de Souza Sales Rocha.pdf: 1422937 bytes, checksum: 58aad769d854c0f77d20fd3b8bf4563f (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2016-07-21T21:05:26Z (GMT) No. of bitstreams: 1 PDF - Eveline Angélica Lira de Souza Sales Rocha.pdf: 1422937 bytes, checksum: 58aad769d854c0f77d20fd3b8bf4563f (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2016-07-21T21:05:36Z (GMT) No. of bitstreams: 1 PDF - Eveline Angélica Lira de Souza Sales Rocha.pdf: 1422937 bytes, checksum: 58aad769d854c0f77d20fd3b8bf4563f (MD5) / Made available in DSpace on 2016-07-21T21:05:36Z (GMT). No. of bitstreams: 1 PDF - Eveline Angélica Lira de Souza Sales Rocha.pdf: 1422937 bytes, checksum: 58aad769d854c0f77d20fd3b8bf4563f (MD5) Previous issue date: 2014-07-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / There is a great demand for new antifungals of different structural classes, acting selectively on new cellular targets with fewer side effects, accordingly medicinal plants are excellent sources for obtaining a wide variety of drugs. This study aimed to investigate the antifungal activity of the plant extract Anadenanthera colubrina (Vell.) Brenan and fitoquimicamente characterize it and set its thermal profile. Shells Anadenanthera colubrina (Vell.) Brenan were collected in the semiarid Paraiba (7º22 '25 "S, 35 ° 59' 32" W), the extract obtained by maceration, using a rotary evaporator and freeze region. For the Determination of Minimum Inhibitory Concentration technique microdilution broth and microorganisms Candida albicans (ATCC 18804), Candida krusei (ATCC 34135), Candida guillermondii (ATCC 6260), Candida parapsilosis (ATCC 22019), Candida tropicalis (ATCC 13803) and clinical strains was used recently isolated from Candida albicans (LM 11, LM 70, LM 410). The Determination of Minimum Fungicidal Concentration was performed by subculture on Sabouraud dextrose agar plates. The action on the cell wall of Candida albicans was investigated by determining the MIC of plant extract in the presence of sorbitol (0.8M), using the microdilution technique. Through scanning electron microscopy, we evaluated the cellular morphology of Candida albicans treated with the MIC of plant extract, the MIC of nystatin suspension (100,000 UI) and free treatment. The combined effect of the two substances (nystatin and vegetable extract Anadenanthera colubrina) studied was determined from the microdilution technique - checkerboard for derivation of Fractional Inhibitory Concentration Index (FIC Index) and by the method of counting viable cells (kinetics growth). In the latter, the following different concentrations of the extract and nystatin products association (CIM; CIM/4; CIM/8) were analyzed. Nonparametric tests Fridman and the Kruskal Wallis test (p <0.05) were performed. In phytochemical screening, we determined the content of total polyphenols, total flavonoids and condensed tannins by spectrophotometry. The thermal profile was traced with the determination of termograviméricas curves (TG) and differential scanning calorimetry (DSC). The toxicity was assessed by quantitation of erythrocyte lysis suffering with different concentrations of plant extract (0.25, 0.5, 1, 2, 4 mg / ml). It was observed that the A.colubrina have fungistatic potential against all tested strains pattern, and fungicidal against recent clinical isolates of Candida albicans. An increase of 1 mg/ml to 8mg/ml of MIC in the presence of sorbitol suggests that this statement acts by modifying the fungal cell wall. The SEM images show the reduction in the amount of yeast cells after treatment with the plant extract and changes in the fungal cell wall. The FIC index was calculated at 0.375, indicating synergism between the nystatin and the plant extract. Kinetics was observed maintaining the synergism between the products tested over time (p <0.05). 53.18% of total polyphenols were determined, 8.73% from 0.28% condensed tannins and flavonoids. The TG curve A.colubrina extract showed the presence of three stages of thermal decomposition, the more significant weight loss was observed between the extract temperatures and 229,17ºC 657,39ºC with loss of 37.44%. The DSC curves of the extract showed that the thermal processes occur at temperatures between 52,37 °C to 195,5°C. In toxicity testing, none of the tested concentrations became effective cytotoxic concentration 50% (EC50). It is concluded that the plant extract from the bark of Anadenanthera colubrina (Vell.) Brenan is a promising resource in getting a new drug for treatment of oral candidiasis. / Existe uma grande demanda por novos antifúngicos de diferentes classes estruturais, agindo seletivamente sobre novos alvos celulares, com menos efeitos colaterais, nesse sentido as plantas medicinais são excelentes fontes para obtenção de uma grande variedade de drogas. Objetivou-se investigar a atividade antifúngica do extrato vegetal de Anadenanthera colubrina (Vell.) Brenan, bem como caracterizá-lo fitoquimicamente e definir seu perfil térmico. Foram coletadas cascas de Anadenanthera colubrina (Vell.) Brenan na região do semiárido paraibano (7º22' 25" S, 35º 59' 32"W), obtido o extrato através de maceração, rotaevaporação e liofilização. Para a Determinação da Concentração Inibitória Mínima foi utilizada a técnica de Microdiluição em Caldo e os microrganismos Candida albicans, (ATCC 18804), Candida krusei (ATCC 34135), Candida guillermondii (ATCC 6260), Candida parapsilosis (ATCC 22019), Candida tropicalis (ATCC 13803) e cepas clínicas recentemente isoladas de Candida albicans (LM 11, LM 70, LM 410). A Determinação da Concentração Fungicida Mínima foi realizada através de subcultivos em placas contendo ágar Sabouraud dextrose. A ação sobre a parede celular de Candida albicans foi investigada através da determinação da CIM do extrato vegetal na presença de sorbitol (0,8M), utilizando a técnica de microdiluição. Através de Microscopia Eletrônica de Varredura, avaliou-se a morfologia celular de Candida albicans tratada com a CIM do extrato vegetal, a CIM da suspensão de nistatina (100.000UI) e livre de tratamento. O efeito combinado das duas substâncias (nistatina e extrato vegetal de Anadenanthera colubrina) estudadas foi determinado a partir da técnica de microdiluição - checkerboard para derivação do Índice de Concentração Inibitória Fracionada (Índice CIF) e através do método de contagem de células viáveis (cinética do crescimento). Nessa última, foram analisadas as seguintes concentrações variadas do extrato, nistatina e associação dos produtos (CIM/8; CIM/4; CIM). Foram realizados os testes não paramétricos de Fridman e o teste de Kruskal Wallis (p<0,05). Na prospecção fitoquímica, determinou-se o teor de polifenóis totais, de flavonoides totais e de taninos condensados através de espectrofotometria. O perfil térmico foi traçado com a determinação das curvas termograviméricas (TG) e calorimetria exploratória diferencial (DSC). A toxidez foi avaliada através da quantificação de hemácias que sofrem lise celular frente à diferentes concentrações de extrato vegetal (0,25, 0,5, 1, 2, 4 mg/ml). Observou-se que a A.colubrina tem potencial fungistático frente todas as cepas padrão testadas, e, fungicida frente aos isolados clínicos recentes de Candida albicans. O aumento de 1mg/ml para 8mg/ml da CIM na presença de sorbitol sugere que esse extrato age modificando a parede celular do fungo. As imagens em MEV evidenciam a redução na quantidade de células fúngicas após o tratamento com o extrato vegetal e alterações na parede celular fúngica. O índice de FIC calculado foi de 0,375, indicando sinergismo entre a nistatina e o extrato vegetal. Na cinética, observou-se a manutenção do sinergismo entre os produtos testados ao longo do tempo (p<0,05). Foram determinados 53,18% de polifenois totais, 8,73% de taninos condensados e 0,28% de flavonoides. A curva TG do extrato A.colubrina, mostrou a ocorrência de três etapas de decomposição térmica, a perda de massa mais significativa do extrato foi observada entre as temperaturas de 229,17ºC e 657,39ºC, com perda de 37,44%. As curvas DSC do extrato mostraram que os processos térmicos ocorreram no intervalo de temperatura entre 52,37 ºC a 195,52ºC. No teste de toxidez, nenhuma das concentrações testadas chegou a ser a concentração citotóxica efetiva 50% (EC50). Conclui-se que o extrato vegetal da casca de Anadenanthera colubrina (Vell.) Brenan é um recurso promissor na obtenção de um novo fármaco para tratamento de candidose oral.