Submitted by Isabela Jacob Moro null (isabela.jm87@gmail.com) on 2016-07-22T18:18:24Z No. of bitstreams: 1 Dissertacao de Isabela Jacob Moro_corrigida.pdf: 4133116 bytes, checksum: e046f271d663b557edcdc98a5fc6eb8f (MD5) / Rejected by Ana Paula Grisoto (grisotoana@reitoria.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo a orientação abaixo: O arquivo submetido não contém a folha de aprovação e está sem a ficha catalográfica. A versão submetida por você é considerada a versão final da dissertação/tese, portanto não poderá ocorrer qualquer alteração em seu conteúdo após a aprovação. Corrija esta informação e realize uma nova submissão contendo o arquivo correto. Agradecemos a compreensão. on 2016-07-22T20:15:50Z (GMT) / Submitted by Isabela Jacob Moro null (isabela.jm87@gmail.com) on 2016-07-22T20:54:52Z No. of bitstreams: 1 Dissertacao de Isabela Jacob Moro_corrigida.pdf: 4201818 bytes, checksum: 6037a407d0b6213632ec0c0218c59ecc (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-07-28T19:24:20Z (GMT) No. of bitstreams: 1 moro_ij_me_arafcf.pdf: 4201818 bytes, checksum: 6037a407d0b6213632ec0c0218c59ecc (MD5) / Made available in DSpace on 2016-07-28T19:24:20Z (GMT). No. of bitstreams: 1 moro_ij_me_arafcf.pdf: 4201818 bytes, checksum: 6037a407d0b6213632ec0c0218c59ecc (MD5) Previous issue date: 2016-06-23 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Produtos derivados de fontes vegetais tem demonstrado seu potencial no combate a diversas doenças, inclusive o câncer, à exemplo dos alcaloides derivados de Catharantus roseus (L.)G. Don, vincristina e vimblastina. A diversidade estrutural das moléculas encontradas em espécies vegetais concebem perspectivas de uso in natura bem como servem de inspiração para o desenvolvimento de outras entidades químicas que serão destinadas ao combate de doenças. O material de estudo pertence a espécie Baccharis trimera (Less.) DC, popularmente conhecida como carqueja e com amplo uso tradicional, principalmente para o alívio de afecções digestivas. O estudo teve por objetivo o fracionamento cromatográfico de derivados vegetais de partes aéreas de B. trimera e a avaliação da citotoxicidade e estudo de morte em linhagens de células humanas normal e tumorais tratadas com o OE, suas frações e componentes, além da flavona eupatorina isolada do extrato acetato de etila. O potencial citotóxico foi avaliado através do método da sulforrodamina B e os estudos de morte através de ensaios de Anexina-V, Hoechst/iodeto de propídeo e ativação de caspase-3. As linhagens celulares humanas utilizadas foram de câncer de mama (MCF-7), hepatocarcinoma (HepG2) e a linhagem de mama normal (MCF-10). As amostras avaliadas foram o óleo essencial, suas frações e componentes (α-humuleno, trans-cariofileno e óxido de cariofileno), além da eupatorina. Os constituintes majoritários do óleo essencial identificados por CG-EM foram: trans-cariofileno(18,9 %), biciclogermacreno (15,6 %), germacreno D (10,5 %) e δ-cadineno (6,6 %); α-humuleno (2,3 %) e óxido de cariofileno (2,9 %) também foram identificados. As frações do óleo essencial foram obtidas por cromatografia em coluna (sílica gel) e analisadas por CG-DIC e CG-EM, tendo apresentado composições químicas distintas. A flavona eupatorina (pureza em 250 nm de 96 % m/m) foi isolada a partir do extrato acetato de etila utilizando cromatografia em camada delgada preparativa (sílica gel) e identificada por técnicas espectrométricas (EM, UV, RMN de 1H e 13C). Verificou-se o maior potencial de redução de sobrevida com o OE em HepG2 (CI50= 10,40 μg/mL), α-humuleno e trans-cariofileno em MCF7 (CI50 de 7,59 e 11,52 μg/mL, respectivamente) enquanto que a eupatorina mostrou-se mais citotóxica em MCF10A com valor de CI50 pouco distante do encontrado para MCF7 (5,08 e 6,73 μg/mL, respectivamente). Grande parte das frações do OE foi citotóxica às linhagens e foram necessárias concentrações mais altas para atingir o CI50. As amostras apresentaram morte mista em todas as linhagens ocorrendo tanto apoptose quanto necrose. Não houve significância estatística no ensaio de ativação da via de apoptose por caspase-3. Constatou-se a eficiência das técnicas cromatográficas no tocante à identificação e separação de constituintes químicos, das técnicas espectrométricas para a identificação e determinação estrutural da substância eupatorina, bem como o potencial de morte celular, e até mesmo antitumoral, de amostras analisadas neste trabalho. / Natural products diversity is an important source to find potential treatments against various diseases, including cancer. Vincristine and vimblastine, alkaloids derived from Catharantus roseus (L.) L. Don, are some of the examples. The structural diversity of molecules from plant species allow the in natura usage and also the development of other chemical entities that will be used to diseases control. Baccharis trimera (Less.) DC., popularly known as carqueja, has an extensive traditional use, especially for digestive disorders. The aims of this study were the chromatographic fractionation of aerial parts from carqueja, in vitro evaluation of the cytotoxic potential through sulforhodamine B assay and cellular death by Annexin-V assay, Hoechst/propidium iodide method and caspase-3 activation, in normal and tumor cell lines treated with essential oil, its fractions and components, and the flavone eupatorin isolated from ethyl acetate extract. The cell lines used were human breast cancer (MCF-7), hepatocellular carcinoma (HepG2) and normal breast line (MCF-10a). The evaluated samples were: essential oil, its fractions and components (α-humulene, β- caryophyllene and caryophyllene oxide) and eupatorin. The major constituents of the essential oil identified by GC-MS were: β-caryophyllene (18.9%), bicyclogermacrene (15.6%), germacrene D (10.5%), δ-cadinene (6.6%). Other components of the essential oil, such as α-humulene (2.3%) and caryophyllene oxide (2.9%) were also identified. The essential oil fractions were obtained by column chromatography (silica gel) and analyzed by GC-FID and GC-MS, presenting different chemical compositions. The flavone eupatorin (96% purity in 250 nm; w/w) was isolated from ethyl acetate extract using preparative thin layer chromatography (silica gel) and identified by spectrometric methods (MS, UV, 1H and 13C NMR). The largest decrease in cell viability for HepG2 was observed with essential oil (IC50 = 10.40 μg / mL); α-humulene and trans-caryophyllene for MCF7 cells (IC50 7.59 and 11.52 μg / mL, respectively) and eupatorin for MCF10A (IC50= 5.08 μg / mL). IC50 for eupatorin in MCF7 cells was 6.73 μg / mL. Most of the essential oils fractions were cytotoxic to the cell lines and higher concentrations were required to achieve IC50. There was not a cell death standard since apoptosis and necrosis occurred as well. Statistical significance was not found in caspase-3 assay. Chromatographic techniques were efficient for compounds identification and separation, and spectrometry techniques for structural determination of eupatorin. The results from this research shown therapeutic potential against cancer for the studied samples. / FAPESP: 2014/10552-3

Made available in DSpace on 2015-05-14T12:59:51Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2644310 bytes, checksum: 2bb61cfd68e68c24b1d4a31bbf372a45 (MD5) Previous issue date: 2008-12-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The use of plants as a treatment method has gained prominence and became fashionable in the world. In Brazil is not different. In recent years some brazilian states has been implementing phytotherapy as an alternative therapy for the public health. However there are some drawbacks as the lack of scientific information that supporting traditional knowledge of some species. The Licania macrophylla Benth species belongs to the Chrysobalanaceae family and is known popularly as "anauera" or "anuera." The stem bark of this species is used in the state of Amapá like amebicide as antidiarrheal. The Pradosia huberi Ducke species belongs to the Sapotaceae family, and is popularly called "fresh bark" or "stick candy ' and its stem bark is used by Amazonian peoples to assist in the treatment of stomach problems and indigestion. However the use of the stem bark of these species in the preparation of herbal formulation or even home preparation is a predatory practice, because recovery of the damage caused by removal of the bark takes years to rebuild, if not eliminates the specimen. The aim of this work was the phytochemical and biological study of the Licania macrophylla Benth and Pradosia huberi (Ducke) Ducke species and compare the chemical composition of leaves and stem bark of both species, so as to suggest that you can replace the use of shells stem by the leaves, and thus contribute to the conservation of the species under discussion. The species studied were collected at Porto Grande - Amapá - Brazil and specimens are deposited in the Herbarium Amapaense HAMAB IEPA. The phytochemical study of the stem bark of the L. macrophyla Benth (Chrysobalanaceae) species led to the isolation of (-)-4'-O-methyl-epigallocatechin-3'-O-α- L-raminosídeo (LM-1), (-)-4'-methyl-epigallocatechin (Lm -2), while the leaves were isolated pheophytin A (Lm-3), 132-hydroxy-(132-S) pheophytin A (Lm-4), pheophytin B (Lm-5), β-sitosterol (Lm-6a ), stigmasterol (Lm-6b), β-sitosterol-O-glycoside (Lm-7), alcohol betulínco (Lm-8) and oleanolic acid (Lm-9), the first not reported in the literature. From species was isolated Pradosia huberi 2,3-dihidromiricetina-3-α-LO-raminosídeo (Ph-1) and leaves the fatty ester erythrodiol (Ph-2), ester of fatty oleanolic acid (Ph-3) fatty ester betulínco acid (Ph-4) and espinasterol (Ph-5), all identified by techniques 1H NMR and 13C single and two-dimensional and comparisons with published literature. Evaluation of antidiarrhoeal activity of the methanol extract of the stem bark (EMC) of L. macrophylla showed that this does not interfere with intestinal parameters: modulation of normal defecation, cathartic agent-induced diarrhea and intestinal transit stimulated. But EMC had to be active against Staphylococcus aureus ATCC 25928, Pseudomonas aeruginosa ATCC 25853 and Escherichia coli ATCC 10536, a result similar to that seen with Lm-1. The methanol extract of leaves (EMF) of L. macrophylla was active against the twelve bacterial strains tested, a result similar to that observed with Lm-4 and Lm-9. Regarding the chemical constituents isolated from the stem bark and leaves in two species can state that are not similar. Regarding the antimicrobial activity of EMC and EMF were similar, suggesting that the activity is antidiarrhoeal proven that one should only antimicrobial can be no substitution of the stem bark of the leaves. / O uso das plantas como recurso terapêutico tem ganhado destaque e tornado modismo no mundo. No Brasil não é diferente, e nos últimos anos alguns estados da federação vem implantando a fitoterapia como alternativa terapêutica para o sistema único de saúde. Porém algumas dificuldades são encontradas, entre elas a falta de informações científicas que subsidie o conhecimento tradicional de determinadas espécies. A espécie Licania macrophylla Benth pertence a família Chrysobalanaceae e é conhecida popularmente por anauera ou anuera . As cascas do caule dessa espécie é usada no estado do Amapá como antidiarréica e amebicida. A espécie Pradosiahuberi Ducke, pertence à família Sapotaceae, e é designada popularmente de casca doce ou pau doce , suas cascas do caule são usadas pelos povos amazônicos como auxiliar no tratamento de problemas gástricos e má digestão. No entanto o uso das cascas do caule dessas espécies na preparação de fitoterápico ou mesmo em preparações caseiras é uma prática predatória, pois a recuperação do dano causado pela retirada das cascas leva anos para sua reconstituição, quando não elimina o espécime. O objetivo deste trabalho foi realizar o estudo fitoquímico e biológico das espécies Licania macrophylla Benth e Pradosia huberi (Ducke) Ducke e comparar a composição química entre folhas e cascas do caule das duas espécies, de forma a sugerir se é possível substituir o uso das cascas do caule pelas folhas, e assim contribuir com a conservação das espécies em discussão. As espécies estudadas foram coletadas no município de Porto Grande Amapá Brasil e exsicatas estão depositadas no Herbário Amapaense HAMAB do IEPA. O estudo fitoquímico das cascas do caule da espécie L. macrophyla Benth (Chrysobalanaceae) levou ao isolamento de (-)- 4 -O-metil-epigalocatequina-3 -O-α-L-raminosídeo (Lm-1), (-)-4 -metil-epigalocatequina (Lm-2), enquanto das folhas foram isoladas feofitina A (Lm-3), 132-hidroxi-(132-S)- feofitinaA (Lm-4), feofitina B (Lm-5), β-sitosterol (Lm-6a), estigmasterol (Lm-6b), β-Oglicosídeo- sitosterol (Lm-7), álcool betulínco (Lm-8) e ácido oleanólico (Lm-9), sendo o primeiro não relatado na literatura. Da espécie Pradosiahuberi foi isolado 2,3- dihidromiricetina-3-α-L-O-raminosídeo (Ph-1) e das folhas éster graxo do eritrodiol (Ph- 2), éster graxo do ácido oleanólico (Ph-3), éster graxo do ácido betulínco (Ph-4) e espinasterol (Ph-5), todas identificadas através de técnicas de RMN de 1H e 13C uni e bidimensionais e comparações com a literatura. A avaliação da atividade antidiarréica do extrato metanólico das cascas do caule (EMC) de L. macrophylla mostrou que esse não interfere nos parâmetros intestinais: modulação da defecação normal; diarréia induzida por agente catártico e trânsito intestinal estimulado. Porém EMC apresentou-se ativo frente à Staphylococcus aureus ATCC 25928, Pseudomonas aeruginosa ATCC 25853 e Escherichia coli ATCC 10536, resultado semelhante ao obtido com Lm-1. O extrato metanólico das folhas (EMF) de L. macrophylla mostrou-se ativo frente as doze cepas bactérianas testadas, resultado semelhante ao observado com Lm-4 e Lm-9. No que se refere aos constituintes químicos isolados das cascas do caule e folhas nas duas espécies pode-se afirmar que não são semelhantes. Já em relação a atividade antimicrobiana do EMC e EMF esses apresentaram-se semelhantes, sugerindo que se a atividade antidiarréica fica comprovada que se deve apenas a ação antimicrobiana pode-se haver substituição do uso das cascas do caule pela folhas.

O Hibiscus rosa-sinensis L. e o Hibiscus syriacus L., da família Malvaceae são utilizados na área ornamental, mas nos últimos anos vem ganhando espaço na área alimentícia como flores comestíveis. Alguns estudos demonstram o potencial antibacteriano destas variedades frente a diversos microrganismos e sobre sua composição nutricional e fitoquímica há poucas pesquisas. Este trabalho teve por objetivo analisar a intensidade de atividade de inibição (IINIB) e a inativação bacteriana (IINAB) in vitro dos dois extratos alcoólicos das flores dos hibiscos e a relação com os polifenóis e antocianinas, e quantificar os compostos nutricionais e bioativos comparados com a atividade antioxidante. Avaliou-se a ação antibacteriana frente às bactérias de interesse alimentar, Staphylococcus aureus e Salmonella Enteretidis resultando em diferenças significativas entre as médias dos valores arbitrários (IINIB/IINAB). Observou-se a resistência da primeira bactéria com a segunda respectivamente em ambos os extratos vegetais. O doseamento dos compostos fitoquímicos presentes constatou que as plantas possuem correlação com a atividade antibacteriana e as propriedades químicas demonstraram valores significativos do ponto de vista nutricional e detectou-se efeito relevante com a atividade antioxidante. / The Hibiscus rosa-sinensis L. and Hibiscus syriacus L., Malvaceae the family are used in the ornamental area, but in recent years has been gaining ground in the food area as edible flowers. Some studies have demonstrated the antibacterial potential of these varieties against various microorganisms and on their nutritional composition and phytochemical little research. This study aimed to analyze the intensity of activity inhibition (IINIB) and bacterial inactivation (IINAB) in vitro of both alcoholic extracts of petals of flowers of hibiscus and relationship with polyphenols and anthocyanins, and quantify the nutritional and bioactive compounds compared with antioxidant activity. We evaluated the antibacterial action on the bacteria of food interest, Staphylococcus aureus and Salmonella Enteretidis resulting in significant differences between the means of arbitrary values (IINIB/IINAB), where there was resistance from the first to the second bacterium in both extracts vegetables. The determination of phytochemical compounds found that plants have a correlation with the antibacterial activity and chemical properties showed significant amounts of nutritional standpoint and significant effect was detected with antioxidant activity.

O Hibiscus rosa-sinensis L. e o Hibiscus syriacus L., da família Malvaceae são utilizados na área ornamental, mas nos últimos anos vem ganhando espaço na área alimentícia como flores comestíveis. Alguns estudos demonstram o potencial antibacteriano destas variedades frente a diversos microrganismos e sobre sua composição nutricional e fitoquímica há poucas pesquisas. Este trabalho teve por objetivo analisar a intensidade de atividade de inibição (IINIB) e a inativação bacteriana (IINAB) in vitro dos dois extratos alcoólicos das flores dos hibiscos e a relação com os polifenóis e antocianinas, e quantificar os compostos nutricionais e bioativos comparados com a atividade antioxidante. Avaliou-se a ação antibacteriana frente às bactérias de interesse alimentar, Staphylococcus aureus e Salmonella Enteretidis resultando em diferenças significativas entre as médias dos valores arbitrários (IINIB/IINAB). Observou-se a resistência da primeira bactéria com a segunda respectivamente em ambos os extratos vegetais. O doseamento dos compostos fitoquímicos presentes constatou que as plantas possuem correlação com a atividade antibacteriana e as propriedades químicas demonstraram valores significativos do ponto de vista nutricional e detectou-se efeito relevante com a atividade antioxidante. / The Hibiscus rosa-sinensis L. and Hibiscus syriacus L., Malvaceae the family are used in the ornamental area, but in recent years has been gaining ground in the food area as edible flowers. Some studies have demonstrated the antibacterial potential of these varieties against various microorganisms and on their nutritional composition and phytochemical little research. This study aimed to analyze the intensity of activity inhibition (IINIB) and bacterial inactivation (IINAB) in vitro of both alcoholic extracts of petals of flowers of hibiscus and relationship with polyphenols and anthocyanins, and quantify the nutritional and bioactive compounds compared with antioxidant activity. We evaluated the antibacterial action on the bacteria of food interest, Staphylococcus aureus and Salmonella Enteretidis resulting in significant differences between the means of arbitrary values (IINIB/IINAB), where there was resistance from the first to the second bacterium in both extracts vegetables. The determination of phytochemical compounds found that plants have a correlation with the antibacterial activity and chemical properties showed significant amounts of nutritional standpoint and significant effect was detected with antioxidant activity.

Made available in DSpace on 2014-07-29T15:16:30Z (GMT). No. of bitstreams: 1 dissert2e3_schliewe.pdf: 95832 bytes, checksum: 9e10097684779e5ba25e91fcdc7796bb (MD5) Previous issue date: 2008-06-26 / Eriope with about 30 species is a genus of the family Lamiaceae, subfamily Nepetoideae, tribe Ocimae and subtribe Hyptidinae, out of this 20 species are restricted to the campos rupestres of the Brazil. In Goiás, the Chapada dos Veadeiros, Serra de Caldas, Serra dos Cristais, Serra Dourada and Serra dos Pireneus are typical rupestres environments. Beyond these places, in environments of cerrado, in the cities of Colinas do Sul, Goiânia, Mineiros, Planaltina de Goiás, Uruaçu and Vianópolis were located specimens of Eriope. The aim of this work is to supply new morphologic, anatomical and phytochemistry data to contribute with the taxonomy of Eriope. Eight taxa were collected: Eriope complicata Mart. ex Benth., Eriope crassipes Benth., Eriope crassipes Benth. subsp. crassipes var. acutifolia Benth., Eriope crassipes Benth. subsp. crassipes var. macrophylla Benth., Eriope crassipes Benth. subsp. cristalinae Harley, Eriope foetida A. St. Hil. ex Benth., Eriope velutina Epling and one taxon that probaly it s a new species endemic of Cristalina-GO region. The subspecies Eriope crassipes Benth. subsp. cristalinae Harley presented morphologic and anatomical characteristics differential among group, in this way the suggestion is the manutetion specific status Eriope cristalinae (Harley) C. T. Rizzini and in synonim Eriope crassipes Benth. subsp. cristalinae Harley. Identification keys were elaborate using morphologic and anatomical data for the taxa in study. The species presented anthraquinone flavonic and saponin heterosides; triterpenes and fenolic compound. In the essential oils 58 compounds were identificated, α-copaen, Aromadendrene and copaen-4α-ol were identificated in all the taxa. Key words: morpho-anatomical, / O gênero Eriope com cerca de 30 espécies, pertence à família Lamiaceae, subfamília Nepetoideae, tribo Ocimeae e subtribo Hyptidinae, dessas 20 espécies são restritas aos campos rupestres do Brasil. Em Goiás, a Chapada dos Veadeiros, Serra de Caldas, Serra dos Cristais, Serra Dourada e a Serra dos Pireneus, são ambientes rupestres, nos quais ocorrem espécies desse gênero. Além destes locais, em ambientes de cerrado, nos municípios de Colinas do Sul, Goiânia, Mineiros, Planaltina de Goiás, Uruaçu e Vianópolis foram localizados espécimes de Eriope. Este trabalho objetiva fornecer dados morfológicos, anatômicos e fitoquímicos visando contribuir com a taxonomia de Eriope. Foram coletados oito taxa: Eriope complicata Mart. ex Benth., Eriope crassipes Benth., Eriope crassipes Benth. subsp. crassipes var. acutifolia Benth., Eriope crassipes Benth. subsp. crassipes var. macrophylla Benth., Eriope crassipes Benth. subsp. cristalinae Harley, Eriope foetida A. St. Hil. ex Benth. e Eriope velutina Epling e um taxon, Eriope sp., que provavelmente trata-se de uma nova espécie endêmica na região de Cristalina-GO. A subespécie Eriope crassipes Benth. subsp. cristalinae Harley apresentou características morfológicas e anatômicas distintas dentro do grupo, desse modo sugere-se manter o status específico Eriope cristalinae (Harley) C. T. Rizzini e como sinonímia Eriope crassipes Benth. subsp. cristalinae Harley. Foram elaboradas chaves de identificação usando dados morfológicos e anatômicos para os taxa em estudo. As espécies estudadas apresentaram: Heterosídeos antraquinônicos, flavonóides e saponínicos; triterpenos e compostos fenólicos. Em relação aos óleos essenciais dos 58 componentes identificados, α-copaeno, Aromadendreno, β- copaeno α-4-ol são comuns a todos os taxa.

Made available in DSpace on 2016-08-19T17:47:05Z (GMT). No. of bitstreams: 1 DISERTACAO_GIZELI SANTOS LOURENCO COUTINHO.pdf: 2757714 bytes, checksum: bc543fd210466bf4c5a1fd09ff430d3c (MD5) Previous issue date: 2013-06-10 / Himatanthus drasticus (janaúba) is a tree with dense foliage at the ends of branches, oblique and ovals leaves, subcoriaceous, shiny, glabrous, dark green, with apex rounded to obtuse, short petioles and rough bark that exudes a white latex widely used in popular medicine in treatment of tumors, worms, gastritis, arthritis, infertility and also cancer. Therefore, this study aims to do an ethnopharmacological survey, determine the phytochemical and metal compositions, and to evaluate the antioxidant and microbiological activity of the hydroalcoholic extract from the leaves, bark and latex of the plant species Himatanthus drasticus. Qualitative tests were conducted for detection of secondary metabolites, and mineral detection was performed by atomic absorption (ICP-OES technique). In relation to the antioxidant activity was used in vitro method with DPPH and for antimicrobial activity have been used standard strains (ATCC) of gram-positive bacteria (Staphylococcus aureus, Streptococcus agalacicus), gram-negative (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis) and yeasts of Candida gender (C. albicans, C. parapsilosis, C. tropicallis, C.glabrata) through tests in diffusion in the solid and in agar diffusion, determining the Minimum Inhibitory Concentration (MIC). Regarding the popular use in Maranhão is indicated for: inflammation, inflammation on uterus, gastritis, ulcers, blood cleanser, prostate, cancer, as restorative and against infertility. Were found in the leaves of Himatanthus drasticus flavonoides, condensed tannins, alkaloids, steroid free, triterpenes, saponins, resins, coumarins. In barks, were found strongly positive results for anthocyanins, flavonoids and anthocyanidins. In the bark and latex were detected the presence of steroids and triterpenoids. Still in the bark was found a great percentage of calcium, potassium and magnesium, in the latex was observed higher percentage of calcium, magnesium and in the leaves the predominance of minerals like calcium, potassium, phosphorus and magnesium. The hydroalcoholic extract of the bark and leaves of H. drasticus showed significant antioxidant activity with EC50% 27.8848 9g / mL and CE50 of 31.62 9g / mL, respectively. In vitro testing was observed inhibition of growth of the tested strains with hydroalcoholic extract from the bark and leaves of H. drasticus, but the latex was inactive. Only the latex has caused cell death in the title 1:16 in carcinoma larynx cell (HEP-2) and 1:8 in uterine carcinoma cell (Hela). Therefore, further studies should be conducted to better scientific understanding of the species H. drasticus / Himatanthus drasticus (janaúba) é uma árvore com folhagem densa nas extremidades dos ramos, folhas obovais, ubcoriáceas, brilhantes, glabras, verde escuro, com ápice arredondado a obtuso, pecíolos curtos e casca rugosa que exsuda um látex branco bastante utilizado na medicina popular no tratamento de tumores, verminoses, gastrites, artrites, infertilidade e também contra o câncer. Portanto, este trabalho tem como objetivo fazer um levantamento etnofarmacológico da utilização da espécie vegetal pela população ludovicense, determinar a composição fitoquímica e de metais, além de avaliar a atividade antioxidante e microbiológica do extrato hidroalcoólico das folhas, casca e látex da espécie vegetal H. drasticus. Realizaram-se testes qualitativos para detecção de metabólitos secundários e a detecção de minerais foi realizada por absorção atômica (técnica ICP-OES). Em relação à atividade antioxidante utilizou-se o método in vitro com DPPH e para a atividade antimicrobiana foram utilizadas cepas padrão (ATCC) de bactérias gram-positivas (Staphylococcus aureus, treptococcus agalacicus), gramnegativas (Pseudomonas aeruginosa, escherichia coli, Klebsiella pneumoniae, Proteus mirabilis) e leveduras do gênero Candida (C. albicans, C. parapsilosis, C. tropicallis, C.glabrata) através de testes em difusão no meio sólido e difusão em ágar, determinando-se a Concentração Inibitória Mínima (CIM) e citotoxicidade. Em relação ao uso popular no Maranhão H. drasticus é indicada para: inflamação, inflamação do útero, gastrite, úlceras, depurativo do sangue, próstata, câncer, fortificante e contra infertilidade. Foram encontrados nas folhas de Himatanthus drasticus flavonoides, taninos condensados, alcaloides, esteroides livre, triterpenos, saponinas, resinas, cumarinas. Nas cascas, encontrou-se resultados fortemente positivo para antocianinas, antocianidinas e flavonoides. Tanto na casca quanto no látex foram detectados a presença de esteroides e triterpenóides. Foi encontrado ainda na casca um percentual bastante acentuado de cálcio, potássio e magnésio, no látex se observou maior percentual de cálcio, magnésio e nas folhas a predominância de minerais do tipo cálcio, potássio, fósforo e magnésio. O extrato hidroalcoólico das cascas e folhas de H. drasticus apresentou significativa atividade antioxidante com CE50% 27,8848 Pg/mL e CE50 de 31,62 Pg/mL, espectivamente. No teste in vitro foi observada a inibição de crescimento das cepas testadas com extrato hidroalcoólico das cascas e folhas de H. drasticus, porém o látex não apresentou atividade. Apenas o látex causou morte celular no titulo 1:16 na célula de carcinoma de laringe (HEP-2) e 1:8 na célula de carcinoma uterino (Hela). Por isso, novos estudos devem ser realizados para melhor conhecimento científico da espécie H. drasticus.

Nectandra membranacea (Lauraceae) est une plante très commune aux Petites Antilles. Aucune utilisation traditionnelle de cette plante n’est connue mais le genre Nectandra, qui représente le deuxième plus important de la famille des Lauraceae avec 114 espèces reconnues, présente des propriétés analgésiques, anti-inflammatoires et énergétiques (Le Quesne et al 1980). Nectandra a également démontré une activité antitumorale (Silva-Filho, 2004). Les nombreuses propriétés du genre et le peu de connaissance chimique sur cette espèce nous ont incités à réaliser une étude phytochimique et à évaluer le potentiel biologique des différentes parties de cette plante. L’analyse des lipides a été effectuée, et les principales classes de métabolites secondaires (terpènes, flavonoïdes, alcaloïdes) ont été recherchées dans les feuilles, les écorces et les fruits de Nectandra membranacea. Afin de suivre les variations de la composition chimique en fonction du cycle de vie du végétal, l’étude a été réalisée sur des échantillons (feuilles, écorces et fruits), récoltés à deux stades du cycle de la plante : la germination (stade A) et la fructification (stade B). N. membranacea, grâce à la présence de nombreuses familles chimiques, possède des activités biologiques intéressantes. Les huiles essentielles ont montré des activités anticancéreuses, anti-inflammatoires, antioxydantes et antiherpétiques. Les extraits polyphénoliques issus des différentes parties de la plante ont montré une très grande activité antioxydante (ORAC et DPPH) et in vitro (cellules saines) ainsi que des propriétés anti-inflammatoires notables. Les extraits d’alcaloïdes totaux issus des trois parties ont été évalués pour la maladie d’Alzheimer et ont révélé de très bons effets neuroprotecteurs, plus particulièrement pour les extraits de feuilles récoltées lors de la germination. Ce travail a permis d’élargir la connaissance des huiles essentielles de l’espèce (composition pour plusieurs parties et activité biologique). Des flavonoïdes ont été identifiés, mais la plante s’est révélée particulièrement riche en alcaloïdes (une quinzaine) de type isoquinoliques et indoliques. N. membranacea se révèle être une plante aux nombreuses propriétés, au regard de ses activités biologiques intéressantes. Des extraits pourraient être utilisés comme principes actifs dans des médicaments mais également en industrie cosmétique. / Nectandra membranacea (Lauraceae) is a very common plant in the French West Indies. No traditional use of this plant is known, but the genus Nectandra, which represents the second most important of Lauraceae family with 114 recognized species, has got analgesic, anti-inflammatory and energizing activities (Le Quesne et al 1980). Nectandra was also identified as a potential antitumor agent (Silva-Filho, 2004). Many properties of the genus, and some chemical knowledge of this species, have prompted us to perform phytochemical and biological study of different parts of the plant. Lipids and the main classes of secondary metabolites (terpenes, flavonoids, alkaloids) were investigated in leaves, bark and fruits of Nectandra membranacea. To determine the monitoring changes in the chemical composition, according to the life cycle of the plant, the study was carried out on samples (leaves, bark and fruits) harvested at two stages of the cycle plant : germination (stage A) and fruit stage (stage B). N. membranacea, thanks to the presence of many chemical families, has interesting biological activities. Essential oils have shown anticancer, anti-inflammatory, antioxidant activities. They revealed antiviral activity on the herpes virus. The polyphenolic extracts from different parts of the plant showed a high antioxidant activity (ORAC and DPPH tests) and in vitro (healthy cells) and significant anti-inflammatory properties. The total alkaloids extracted from the three parts were evaluated for Alzheimer’s disease and were found to have very good protective effects, especially the leaves’ extracts collected during germination. This work has expanded knowledge of the essential oil of the species (composition of several parts and biological activity). Flavonoids have been identified, but the plant was shown to be particularly rich in alkaloids (fifteen were identified) of indolic and isoquinolic type. N. membranacea presents many properties. In view of its interesting biological activities, it could be used for many drugs but also in cosmetic area.

Avec la prise de conscience de l'impact environnemental et sanitaire des pesticides et l'apparition de phénomènes de résistance face à ces produits, les exigences règlementaires européennes (UE 1107/2009) encadrant leur mise sur le marché sont de plus en plus drastiques. Pour répondre à ces exigences, de nouveaux types de produits de protection des cultures font leur apparition : les bio-pesticides, ou produits de biocontrôle. Il s'agit de produits naturels d'origine végétale, animale ou microbienne, à priori moins rémanent et impactant pour l'environnement que les pesticides conventionnels. C'est dans ce contexte que mes travaux de thèse ont eu pour but le développement de biopesticides d'origine végétale destinés à lutter contre les champignons pathogènes des plantes, pour lesquels il existe encore peu ou pas de moyens de lutte naturels. Au cours de cette thèse, j'ai pu sélectionner à travers différents criblages, écologique, économique et biologique deux extraits végétaux efficaces sur des cibles biologiques d'intérêt et présentant une toxicité sur organismes non cibles bien plus faible que des fongicides conventionnels. Les molécules responsables de l'activité ont été identifiées par fractionnement bio-guidé et analyses spectrales. L'activité des extraits a pu être validée en champs notamment sur mildiou de la vigne (Plasmopara viticola) et moniliose de la nectarine (Monilinia fructigena) après une optimisation de leur formulation. / With the awareness of the environmental and sanitary impact of pesticides and emergence of resistance phenomena, the European regulation requirements (EU 1107/2009) supervising their marketing authorization are more and more drastic. To meet these requirements, new phytoprotection products are emerging: biopesticides also called biocontrol products. They are natural product of plant, animal or microbial origins, likely to be less persistent and more environmentally friendly than conventional pesticides.It is against this background that my thesis work aimed to develop some biopesticides from plant origin, for protection against phytopathogenic fungi for which there are no or few natural product.In this work, I selected, by ecological, economical and biological screening, two plant extracts, effective against biological targets and far less toxic against non-target organism than tested conventional pesticides.Active molecules were identified by bioguided fractionation and spectral analysis. Extracts activities have been evaluated in field test against vine downy mildew (Plasmopara viticola) and Moniliose infection (Monilinia fructigena)

L'objectif de cette thèse est d'évaluer les modifications du métabolisme secondaire des plantes contaminées aux éléments trace métalliques (ETM) et leurs conséquences sur les communautés bactériennes rhizosphériques associées incluant des bactéries présentant des phénotypes de MultiDrug Résistance (MDR). Nous nous sommes focalisés sur deux contextes de sols exposés aux métaux : la phytoremédiation de sites miniers au Vietnam et la reconversion de sols agricoles contaminés par la re-déposition atmosphérique d'activités métallurgiques en France. Nos résultats ont mis en évidence que la contamination par différents types de métaux (dont Cu et Pb principalement) a conduit à une altération de la production des métabolites secondaires des racines, tiges et feuilles de la plante hyperaccumulatrice Pteris vittata et que concernant les racines des tendances similaires dans les changements métaboliques ont pu être observés dans un autre type de contexte de pollution (Zn et Pb plus particulièrement). De même, les profils métaboliques des parties souterraines (racines et rhizomes) de Miscanthus x giganteus ont été modifiés par les concentrations en Pb, Cd et Zn des sols agricoles. Pour les deux plantes examinées des dérivés de l'acide chlorogénique ont été retrouvés en proportions augmentées dans les racines malgré des contextes de nature des sols et de pollutions métalliques très contrastés. Cependant, les dérivés de tanin catéchiques sont spécifiquement trouvés en proportions plus élevées dans les racines de P. vittata sous pression métallique. Ces polyphénols sont connus pour leur capacité à piéger les radicaux libres et leur pouvoir antioxydant et pourraient donc être impliqués dans l'adaptation de ces plantes au stress métallique en contribuant à limiter le stress oxydatif généré par les ETM. Au niveau des parties aériennes, nous n'avons étudié que le changement pour P. vittata et avons mis en évidence une proportion plus élevée de dérivés flavonoïdiques pour les plantes contaminées. Nos résultats de métagénomique nous permettent de conclure également sur un effet des ETM sur la diversité et la richesse spécifique des communautés bactériennes des sols étudiés : une forte contamination en Cu (10 fois la limite autorisée) a diminué la diversité et la richesse bactérienne, alors que pour des niveaux en ETM plus modérés incluant Cu, Pb et Zn, la diversité des communautés bactériennes rhizosphériques semble plus influencée par la plante ou la saison plutôt que par l'effet des ETM. Cet effet sur la composition bactérienne de la rhizosphère de P. vittata se traduit par un enrichissement de certains genres connus comme pathogènes opportunistes de l'homme, notamment Ralstonia, Acinetobacter, Burkholderia et Mycobacterium. En outre, le genre Cupriavidus, connu comme très résistant aux ETM est le seul genre spécifiquement associé à P. vittata qui ait été augmenté au sein de la communauté rhizosphérique pour les deux sites miniers étudiés par rapport aux sols rhizosphériques non pollués. Ce genre pourrait donc être impliqué dans le processus d'adaptation de cette plante au stress métallique. Quant aux communautés rhizosphériques de Miscanthus x giganteus, la sélection de Stenotrophomonas et Pseudomonas dans les sols agricoles contaminés a été observée. Dans le cadre de cette thèse nous avons également mis au point une méthode rapide pour tester l'impact de métabolites végétaux sur des souches pathogènes d'origine clinique et environnementale et également évaluer leur activité inhibitrice de pompes à efflux (IPE) de la famille des RND. Nos données ont ainsi permis de mettre en évidence des activités intéressantes et comparables à celle de l'inhibiteur de pompe à efflux PAßN pour des composés testés qui étaient extraits des racines de Fallopia x bohemica ou des dérivés de ces derniers. / The objective of this thesis is to evaluate the modification of plant secondary metabolism production contaminated with metallic trace elements (MTE) and its consequences on the associated rhizospheric bacterial communities including bacteria presenting MultiDrug Resistant (MDR) phenotypes. We have focused on two contexts of metals exposure: the phytoremediation of mining sites in Vietnam and the reconversion of agricultural soils contaminated by the atmospheric re-deposition of metallurgical activities in France. Our results highlighted that contamination by different types of metals (mainly Cu and Pb) has led to an alteration in the production of secondary metabolites in the roots, stems and leaves of the hyper-accumulating Pteris vittata and for roots, a similar trend in the metabolic changes could be observed in another type of pollution context (Zn and Pb more particularly). Similarly, the metabolic profiles of the underground parts (roots and rhizomes) of Miscanthus x giganteus were modified by the concentrations of Pb, Cd and Zn in agricultural soils. For the two plants examined chlorogenic acid derivatives have been found in increased proportions in the roots despite soil type and pollution context were highly contrasted. However, catechic tannin derivatives are specifically found in higher proportions in the roots of P. vittata under metal pressure. These polyphenols are known for their ability to scavenge free radicals and their antioxidant properties and thus could be involved in the adaptation of these plants to metallic stress by helping to limit the oxidative stress generated by MTE. At the level of the aerial parts, we studied only the change for P. vittata and evidenced higher proportions of flavonoid derivatives for contaminated plants. Our metagenomic results allow us to conclude also on the effect of MTE on the diversity and the specific richness of the bacterial communities of the studied soils: a high contamination of Cu (10 times the allowed limit) decreased dramatically bacterial richness and diversity, while for more moderate MTE levels including Cu Pb and Zn, the diversity of rhizosphere bacterial communities was more explained by plant or season effect rather than an effect of MTE. This effect on P.vittata rhizosphere bacterial composition is reflected by an enrichment in genera known as opportunistic human pathogens, including Ralstonia, Acinetobacter, Burkholderia and Mycobacterium. In addition, Cupriavidus, known as a highly resistant genus, is the only P. vittata specifically associated genus found in increased proportions at both mining sites compared to non-contaminated rhizosphere soils. This genus could then be involved in the adaptation process of this plant with metal stress. As for the rhizospheric communities of Miscanthus x giganteus, the selection of Stenotrophomonas and Pseudomonas in agricultural soils contaminated with MTE was observed. As a part of this thesis, we have also developed a rapid method for testing the impact of plant metabolites on pathogenic strains of clinical and environmental origin and their efflux pump inhibition (EPI) activity of RND family. Our data thus showed interesting and notable EPI activities comparable to that of the efflux pump inhibitor PAßN for tested compounds issued from Fallopia x bohemica roots or for their derivatives.

Ethanol extracts of “jenipapo” (Genipa americana Linnaeus), “umbu" (Spondia tuberosa Arruda), “siriguela” (Spondia purpurea Linnaeus) and “mangaba” (Hancornia speciosa Gomes) were prepared from separate pulp, seeds and peel; except mangaba which are used pulp and peel. The investigation of antioxidant capacity of the ethanol extracts were carried out by the methods of determining the content of ascorbic acid and total phenolics; scavenging 2, 2 -diphenyl-1-picrilhydrazyl and 2, 2’- azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical, antioxidant capacity of reduction of iron and copper and lipid peroxidation using a biomimetic membrane system, and measurement enzymatic of catalase and superoxide dismutase. It was also analyzed the acetylcholinesterase inhibitory activity, cytotoxic effect on corneal epithelial cells of sheep, as well as phytochemicals assays and identification of phenolic compounds and organic acids present in the extracts by UPLC - MS. The highest values of total phenolic content were obtained with peel and seed extracts and to ascorbic acid in the seed of siriguela, showing better antioxidant activities of seeds and peel extracts of siriguela and umbu. Lipid peroxidation assays indicated that genipap pulp is a promising antioxidant. Genipap pulp and siriguela seed ethanol extracts presented an acetylcholinesterase inhibition zone similar to that of the positive control, carbachol. The investigation of phenols and organic acid contents revealed the presence of quercetin (48,38 to 3,88 μg.g-1), quinic acid (43,28 to 41,88 μg.g-1) and citric acid (3,78 to 0,43 μg.g-1) in several extracts and chlorogenic acid with the highest amount found in siriguela seeds (356,93 μg.g-1). These data suggest new uses for these foods as potential antioxidant supplements for the food, pharmaceutical and cosmetic industries. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A partir das cascas, polpas e sementes do jenipapo (Genipa americana Linnaeus), umbu (Spondia tuberosa Arruda), siriguela (Spondia purpurea Linnaeus) e mangaba (Hancornia speciosa Gomes) foram preparados extratos etanólicos, com exceção da mangaba onde o extrato foi preparado utilizando casca e polpa. Os extratos foram submetidos à investigação da capacidade antioxidante através dos métodos de determinação do conteúdo de ácido ascórbico e fenóis totais, sequestro do radical 2,2-difenil-1-picril-hidrazil e 2,2´- azinobis (3-etilbenzotiazolina-6-ácido sulfônico), capacidade antioxidante de redução do ferro e cobre, lipoperoxidação utilizando um sistema de membranas biomimético e mensuração enzimática da catalase e superóxido dismutase. Também foi avaliada a atividade inibitória da enzima acetilcolinesterase, o efeito citotóxico em células epiteliais da córnea de ovelhas, além da realização de ensaios fitoquímicos e a identificação de compostos fenólicos e ácidos orgânicos presentes nos extratos. Os maiores teores de fenóis totais foram obtidos nos extratos das cascas e sementes e no referente ao ácido ascórbico na semente da siriguela, apresentando melhor atividade antioxidante os extratos das sementes e cascas da siriguela e do umbu. Porém, no ensaio de peroxidação lipídica, o extrato etanólico da polpa de jenipapo demonstrou ser um antioxidante promissor. Os extratos etanólicos da polpa do jenipapo e da semente da siriguela apresentaram uma zona de inibição da enzima acetilcolinesterase semelhantes ao controle positivo, carbacol. Na investigação dos compostos fenólicos e ácidos orgânicos por UPLC-MS verificou-se a presença nos extratos de quercetina (48,38 a 3,88 μg.g-1), ácido quínico (43,28 a 41,88 μg.g-1), ácido cítrico (3,78 a 0,43 μg.g-1) em vários extratos. E ácido clorogênico, na semente da siriguela (356,93 μg.g-1). Os resultados obtidos desses extratos sugerem novas formas de utilização desses alimentos como potenciais suplementos antioxidantes na indústria alimentícia, farmacêutica e cosmética.