El objetivo del estudio fue evaluar la composición química del extracto acuoso y etanólico de las hojas frescas de Tagetes elliptica Sm. “chincho” y determinar su actividad antibacteriana, antifúngica in vitro y su aplicación en un alimento andino. La especie vegetal fue obtenida en el centro poblado de Molinos de la provincia de Jauja de la Región Junín. El extracto etanólico se obtuvo por maceración con etanol de 96°y la determinación de la composición química se realizó por el método de Cromatografía de Gases/Espectrómetría de Masas (CG/EM), determinándose los siguientes componentes químicos: dianhydrodulcitol, ceanothine c, resorcinol, pirocatecol, 2-(2-butinil)ciclohexanona, z, z-6,24-tritricontadien-2-ona y 5-hexil-2,4-dimetiloxazol, Se evaluó la actividad antibacteriana y antifúngica in vitro del extracto acuoso demostrándose que no tiene actividad antibacteriana frente a las bacterias: Staphylococcus aureus, Bacillus subtillis, Escherichia coli y Pseudomonas aeruginosa en ninguna de las concentraciones ensayadas de 100, 50 y 25 mg/mL respectivamente. El extracto etanólico de Tagetes elliptica Sm. “chincho” mostró actividad frente a Staphylococcus aureus ATCC 25933 y Pseudomonas aeruginosa ATCC 27853, en las tres concentraciones trabajadas, pero no presenta actividad contra Bacillus subtilis ATCC 6633 y Escherichia coli ATCC 25922. La actividad mostrada frente a Staphylococcus aureus y Pseudomonas aeruginosa no es significativa, ya que a la concentración de 25 mg/mL, el halo de inhibición presentado fue menor de 18 mm. La actividad antifúngica del extracto etanólico frente a Candida albicans ATCC 10231 fue inactivo. El extracto acuoso y etanólico de Tagetes elliptica Smith, debe su fragancia a la composición química que posee su aceite esencial y ejerce actividad antibacteriana debido al resorcinol y pirocatecol; compuestos fenólicos a los que se les atribuiría la actividad antibacteriana, predominando su aroma y sabor en el alimento integrado en el potaje andino denominado pachamanca. / The objective of the study was to evaluate the composition chemistry of aqueous and alcoholic extract and fresh leaves of Tagetes elliptica Sm. ethanolic “chincho” and determine the antibacterial and antifungal activity in vitro and its application in an andean food. The plant species was obtained in the town of mills in the province of Jauja in the Region Junín. The summary was obtained by the method of maceration with ethanol of 96°. The chemical composition is determined by the Mass Spectrometer/gas Chromatograph (GC /MS). The determination of the antibacterial and antifungal activity was determined by the method of diffusion in agar, against the strains of the following microorganisms: Staphylococcus aureus ATCC 25933, Bacillus subtilis ATCC 6633 , Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853; also against the yeast Candida albicans ATCC 10231. With the aqueous extract was its application in an andean food. The analysis GC/MS determined the following chemical components: dianthydrodulcitol, ceanothine c, resorcinol, pirocatecol, 2-(2-butynyl)cyclohexanone, -, z, z-6, 24-tritriacontadien-2one, and 5-hexyl-2,4-dimethyl oxazole, Evaluation of antimicrobial and antifungal activity in vitro of the aqueous extract not showed antibacterial activity against S.aureus, B.subtillis, E.coli and P.aeruginosa at any of the concentrations tested of 100, 50 and 25 mg/mL respectively. Ethanolic extract of Tagetes elliptica Sm. "chincho" showed activity against Staphylococcus aureus ATCC 25933, and Pseudomonas aeruginosa ATCC 27853 worked three concentrations and not having any activity against Bacillus subtilis and Escherichia coli. The activity shown against Staphylococcus aureus and Pseudomonas aeruginosa is not significant, since the concentration of 25 mg/mL, the zone of inhibition presented wasn't more than 18 mm. The antifungal activity of ethanolic and aqueous extract was inactive, the aqueous extract and Tagetes elliptica Sm, ethanolic should its fragrance chemical composition which has its essential oil and antimicrobial activity because of resorcinol and pirocatecol; phenolic compounds that it would endow them antimicrobial activity, dominate aroma and flavor in the andean food integrated into the Andean stew called pachamanca. Key words. Tagetes elliptica Smith, pachamanca, Antibacterial, antifungal.

Made available in DSpace on 2014-06-11T19:25:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-11-24Bitstream added on 2014-06-13T18:26:29Z : No. of bitstreams: 1 biso_fi_me_arafcf.pdf: 1559572 bytes, checksum: 88bf79acbb35f72fdc5b29cab4ad0388 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Universidade Estadual Paulista (UNESP) / Davilla nitida Vahl. e D. elliptica St. Hil (Dilleniaceae) são plantas distribuídas, predominantemente, no Cerrado Brasileiro, utilizadas, na medicina popular, para tratar distúrbios gástricos, intestinais e inchaços, principalmente na região dos testículos e linfonodos. Considerando a importância de investigar os efeitos farmacológicos e toxicológicos de espécies vegetais utilizadas com fins terapêuticos, que podem tornar-se alvo na pesquisa de novos fármacos, foi objetivo do presente trabalho avaliar a atividade fitoestrogênica e genotóxica dessas duas espécies de Davilla, utilizando, para tanto, ensaios de proliferação celular com células MCF-7 de tumor de mama humano, ensaios de mutação gênica reversa com Salmonella typhimurium (teste de Ames) e ensaios com plasmídeo pUC9.1, para analisar quebras nas fitas de DNA. A atividade fitoestrogênica de flavonóides isolados de D. elliptica (quercetina, quercetina-3-O-α-L-rhamnopiranosídeo, miricetina e miricetina-3-O-α-L-rhamnopiranosídeo) não pôde ser propriamente caracterizada devido ao perfil de resposta da linhagem celular MCF-7, utilizada nos ensaios de proliferação. No teste de Ames, os extratos metanólico, hidroalcoólico e a fração acetato de etila de D. nitida, bem como os extratos metanólico, etanólico e clorofórmico de D. elliptica induziram mutações nas linhagens de S. typhimurium. Na presença de cobre, as frações aquosa e acetato de etila das duas plantas, e as substâncias puras, quercetina, quercetina-3- O-α-L-rhamnopiranosídeo, miricetina, miricetina-3-O-α-L-rhamnopiranosídeo e ácido gálico, isoladas de D. elliptica, apresentaram potencial genotóxico, promovendo quebras nas cadeias do DNA, em ensaios realizados com plasmídeo pUC9.1. Tais respostas são compatíveis com a literatura, que relata que algumas classes de metabólitos secundários... / Davilla nitida Vahl. and Davilla elliptica St. Hil (Dilleniaceae) are plants distributed predominantly in Brazilian cerrado region and they are used in popular medicine to treat stomach diseases, diarrhea and swelling, particularly of the lymph nodes and testicles. Considering the importance of investigations about pharmacological and toxic effects of vegetable species used as therapeutic drugs, that may become target on new drugs research, the aim of this work was to evaluate the phytoestrogenic and genotoxic activities of both Davilla species, using human breast tumor MCF-7 cell proliferation assay, Salmonella mutagenicity assay (Ames test) and DNA strand break analysis in pUC9.1 plasmid DNA. Phytoestrogenic activity of flavonoids isolated from D. elliptica (quercetin, quercetin-3-O-α-L-rhamnopyranoside, myricetin and myricetin-3-O-α-Lrhamnopyranoside) could not be determined because of the response profile of MCF-7 cell line used in the proliferation assays. In Ames test, methanolic, ethanolic extracts and ethyl acetate fraction of D. nitida and methanolic, ethanolic and chloroformic extracts of D. elliptica induced mutations in S. typhimurium strains. In the presence of copper, aqueous and ethyl acetate fraction of the plants, and compounds, quercetin, quercetin-3-O-α-Lrhamnopyranoside, myricetin, myricetin-3-O-α-L-rhamnopyranoside and gallic acid, isolated from D. elliptica, showed genotoxic potencial, generating DNA strand breaks in plasmid pUC9.1. Such results are compatible to the literature, which reports that some classes of vegetable secondary metabolites are capable of carrying out mutagenic... (Complete abstract click electronic access below)

A espécie endêmica G. elliptica R. E. Fries não apresentava estudos fitoquímicos e biológicos detalhados na literatura. Assim, o objetivo desse trabalho foi avaliar a composição química e as propriedades biológicas dos óleos essenciais, extratos brutos, alcaloides totais, tortas, frações das tortas, amostras isoladas dessa espécie. O material vegetal foi coletado em Paranapiacaba (Santo André, SP, Brasil). O óleo essencial extraído das folhas por destilação à vapor apresentou um rendimento de 0,2%. A análise histológica das folhas encontrou óleo em células oleíferas localizadas no parênquima esponjoso. A composição do óleo (CG-EM) indicou espatulenol e óxido de cariofileno como compostos majoritários. Os alcaloides totais foram obtidos dos extratos brutos das folhas e dos galhos e analisados por CG-EM, identificando quatro aporfinas (nornuciferina, estefarina, corituberina e asimilobina) e duas protoberberinas (discretamina e caseadina). Os alcaloides totais foram fracionados em coluna cromatográfica ou por Extração em Fase Sólida e purificados por cromatografia em camada preparativa, originando duas amostras (Amostra 9 e 10). Na Amostra 9, foram identificados dois alcaloides aporfinicos nornuciferina e asimilobina (CG-EM e RMN-1H). Na Amostra 10, foram identificados (LC-EM/EM) cinco alcaloides aporfínicos (desidronantenina, glaunidina, liriodenina, N-óxido de oliverina e telikovina) e um alcaloide protoberberínico (caseadina). Caseadina, glaunidina, N-óxido de oliverina e telikovina não foram previamente identificados em Guatteria. Os resíduos dos extratos brutos livres de alcaloides foram fracionados pelo método de partição com solventes de polaridade crescente. Os extratos brutos e as frações acetato de etila e butanólicas de folhas e galhos apresentaram flavonoides (NP-PEG). Nos ensaios biológicos, a melhor atividade antioxidante (sequestro do radical DPPH) foi encontrada para a fração clorofórmica dos galhos (EC50=24,25±1,14 &#181;g/mL) e a torta dos galhos (EC50=26,23±4,20 &#181;g/mL). No ensaio antimicrobiano pelo método turbidimétrico a atividade mais importante foi obtida contra Staphylococcus aureus (ATCC 6538) para os alcaloides totais dos galhos (CIM/CBM=0,12±0,01/0,26 mg/mL) e das folhas (CIM/CBM=0,21±0,01/0,28 mg/mL), e fração hexânica das folhas (CIM/CBM=0,24±0,02/>1 mg/mL). Uma alta atividade antitumoral foi observada frente a células humanas de mama (MCF-7) para Amostra 10 (IC50=2,28±0,18 &#181;g/mL), fração de acetato de etila das folhas (IC50=4,47±0,40 &#181;g/mL), óleo essencial (IC50=7,01±0,23 &#181;g/mL) e os alcaloides totais das folhas (IC50=9,32±0,36 &#181;g/mL). Para as células de próstata (PC-3), foi encontrada atividade para a Amostra 10 (IC50=1,37±0,36 &#181;g/mL) e o óleo essencial (IC50=5,32±0,35 &#181;g/mL). A futura aplicação dos extratos e frações de G. elliptica como um agente antitumoral parece ser segura, pois mantiveram uma viabilidade celular maior do que 90% no ensaio de citotoxicidade com culturas de fibroblasto murino (BALB/c 3T3, ATCC CCL-163) nas concentrações onde a atividade antitumoral foi promissora (<30 &#181;g/mL) contra MCF-7 e/ou PC-3. / The endemic species G. elliptica R. E. Fries had not yet been thouroughly investigated phytochemically and biologically. Therefore, the objective of this work was to evaluated the chemical and biological properties of the G. elliptica essential oils, crude extracts, alcaloid-free extracts, total alcaloids, fractions and isolated samples. The plant material was collected in Paranapiacaba (Santo Andre, SP, Brazil). The leaves essential oil, was obtained by hydrodestillation with a yield of 0.2%. In the leaves, the oil was found in secretory cells, located in the spongy parenchyma. The oil analysis (GC-MS) identified spathulenol and caryophyllene oxide as major compounds. The total alkaloids were extracted from the leaves and branches crude extracts and analysed by GC-MS, identifying four aporphines (nornuciferine, stepharine, corytuberine, assimilobine) and two protoberberines (discretamine and caseadine). Total alkaloids were fractionated by column chromatography or solid phase extraction and purified by preparative thin layer chromatography affording two samples (Sample 9 and 10). In Sample 9, two aporphin alkaloids were found and identified (GC-MS and 1H-NMR) as nornurciferine and assimilobine. In Sample 10, were identified (LC-MS/MS) five aporphin alkaloids (dehydronantenine, glaunidine, liriodenine, N-oliverine oxide and telikovine) and one protoberberine alkaloid (caseadine). Caseadine, glaunidine, oliverine N-oxide and telkovine were not previously reported for Guatteria. The alkaloids-free extracts were fractionated using the increasing polarity solvents. Concerning the biological activities, the highest antioxidant activity (DPPH scavenging) was reported for the branches chloroform fraction (EC50=24.25±1.14 &#181;g/mL) and (EC50=26.23±4.20 &#181;g/mL) alkaloid-free extract. In the turbimetric antimicrobial assay against, the most important activity was found against Staphylococcus aureus (ATCC 6538) for the total alkaloids from branches (MIC/MBC=0.12±0.01/0.26 mg/mL) and leaves (MIC/MBC=0.21±0.01/0.28 mg/mL) and for the leaves hexane fraction (MIC/MBC=0.24±0.02/>1 mg/mL). The highest antitumor activity against human breast cancer cell line (MCF-7) was observed for Sample 10 (IC50=2.28±0.18 &#181;g/mL), leaves ethyl acetate fraction (IC50=4.47±0.40 &#181;g/mL), essential oil (IC50=7.01±0.23 &#181;g/mL) and leaves total alkaloids (IC50=9.32±0.36 &#181;g/mL). For the prostate cancer cell line (PC-3), the most active samples were Sample 10 (IC50=1.37±0.36 &#181;g/mL) and the essential oil (IC50=5.32±0.35 &#181;g/mL). The future application of G. elliptica extracts and fractions as an antitumor agent appears to be safe, because cell viability remained higher than 90% in the cytotoxicity assay with murine fibroblast cultures (BALB/c 3T3, ATCC CCL-163), in the concentration range where the best antitumor activity (<30 &#181;g/mL) against MCF-7 and PC-3 was found.

A espécie endêmica G. elliptica R. E. Fries não apresentava estudos fitoquímicos e biológicos detalhados na literatura. Assim, o objetivo desse trabalho foi avaliar a composição química e as propriedades biológicas dos óleos essenciais, extratos brutos, alcaloides totais, tortas, frações das tortas, amostras isoladas dessa espécie. O material vegetal foi coletado em Paranapiacaba (Santo André, SP, Brasil). O óleo essencial extraído das folhas por destilação à vapor apresentou um rendimento de 0,2%. A análise histológica das folhas encontrou óleo em células oleíferas localizadas no parênquima esponjoso. A composição do óleo (CG-EM) indicou espatulenol e óxido de cariofileno como compostos majoritários. Os alcaloides totais foram obtidos dos extratos brutos das folhas e dos galhos e analisados por CG-EM, identificando quatro aporfinas (nornuciferina, estefarina, corituberina e asimilobina) e duas protoberberinas (discretamina e caseadina). Os alcaloides totais foram fracionados em coluna cromatográfica ou por Extração em Fase Sólida e purificados por cromatografia em camada preparativa, originando duas amostras (Amostra 9 e 10). Na Amostra 9, foram identificados dois alcaloides aporfinicos nornuciferina e asimilobina (CG-EM e RMN-1H). Na Amostra 10, foram identificados (LC-EM/EM) cinco alcaloides aporfínicos (desidronantenina, glaunidina, liriodenina, N-óxido de oliverina e telikovina) e um alcaloide protoberberínico (caseadina). Caseadina, glaunidina, N-óxido de oliverina e telikovina não foram previamente identificados em Guatteria. Os resíduos dos extratos brutos livres de alcaloides foram fracionados pelo método de partição com solventes de polaridade crescente. Os extratos brutos e as frações acetato de etila e butanólicas de folhas e galhos apresentaram flavonoides (NP-PEG). Nos ensaios biológicos, a melhor atividade antioxidante (sequestro do radical DPPH) foi encontrada para a fração clorofórmica dos galhos (EC50=24,25±1,14 &#181;g/mL) e a torta dos galhos (EC50=26,23±4,20 &#181;g/mL). No ensaio antimicrobiano pelo método turbidimétrico a atividade mais importante foi obtida contra Staphylococcus aureus (ATCC 6538) para os alcaloides totais dos galhos (CIM/CBM=0,12±0,01/0,26 mg/mL) e das folhas (CIM/CBM=0,21±0,01/0,28 mg/mL), e fração hexânica das folhas (CIM/CBM=0,24±0,02/>1 mg/mL). Uma alta atividade antitumoral foi observada frente a células humanas de mama (MCF-7) para Amostra 10 (IC50=2,28±0,18 &#181;g/mL), fração de acetato de etila das folhas (IC50=4,47±0,40 &#181;g/mL), óleo essencial (IC50=7,01±0,23 &#181;g/mL) e os alcaloides totais das folhas (IC50=9,32±0,36 &#181;g/mL). Para as células de próstata (PC-3), foi encontrada atividade para a Amostra 10 (IC50=1,37±0,36 &#181;g/mL) e o óleo essencial (IC50=5,32±0,35 &#181;g/mL). A futura aplicação dos extratos e frações de G. elliptica como um agente antitumoral parece ser segura, pois mantiveram uma viabilidade celular maior do que 90% no ensaio de citotoxicidade com culturas de fibroblasto murino (BALB/c 3T3, ATCC CCL-163) nas concentrações onde a atividade antitumoral foi promissora (<30 &#181;g/mL) contra MCF-7 e/ou PC-3. / The endemic species G. elliptica R. E. Fries had not yet been thouroughly investigated phytochemically and biologically. Therefore, the objective of this work was to evaluated the chemical and biological properties of the G. elliptica essential oils, crude extracts, alcaloid-free extracts, total alcaloids, fractions and isolated samples. The plant material was collected in Paranapiacaba (Santo Andre, SP, Brazil). The leaves essential oil, was obtained by hydrodestillation with a yield of 0.2%. In the leaves, the oil was found in secretory cells, located in the spongy parenchyma. The oil analysis (GC-MS) identified spathulenol and caryophyllene oxide as major compounds. The total alkaloids were extracted from the leaves and branches crude extracts and analysed by GC-MS, identifying four aporphines (nornuciferine, stepharine, corytuberine, assimilobine) and two protoberberines (discretamine and caseadine). Total alkaloids were fractionated by column chromatography or solid phase extraction and purified by preparative thin layer chromatography affording two samples (Sample 9 and 10). In Sample 9, two aporphin alkaloids were found and identified (GC-MS and 1H-NMR) as nornurciferine and assimilobine. In Sample 10, were identified (LC-MS/MS) five aporphin alkaloids (dehydronantenine, glaunidine, liriodenine, N-oliverine oxide and telikovine) and one protoberberine alkaloid (caseadine). Caseadine, glaunidine, oliverine N-oxide and telkovine were not previously reported for Guatteria. The alkaloids-free extracts were fractionated using the increasing polarity solvents. Concerning the biological activities, the highest antioxidant activity (DPPH scavenging) was reported for the branches chloroform fraction (EC50=24.25±1.14 &#181;g/mL) and (EC50=26.23±4.20 &#181;g/mL) alkaloid-free extract. In the turbimetric antimicrobial assay against, the most important activity was found against Staphylococcus aureus (ATCC 6538) for the total alkaloids from branches (MIC/MBC=0.12±0.01/0.26 mg/mL) and leaves (MIC/MBC=0.21±0.01/0.28 mg/mL) and for the leaves hexane fraction (MIC/MBC=0.24±0.02/>1 mg/mL). The highest antitumor activity against human breast cancer cell line (MCF-7) was observed for Sample 10 (IC50=2.28±0.18 &#181;g/mL), leaves ethyl acetate fraction (IC50=4.47±0.40 &#181;g/mL), essential oil (IC50=7.01±0.23 &#181;g/mL) and leaves total alkaloids (IC50=9.32±0.36 &#181;g/mL). For the prostate cancer cell line (PC-3), the most active samples were Sample 10 (IC50=1.37±0.36 &#181;g/mL) and the essential oil (IC50=5.32±0.35 &#181;g/mL). The future application of G. elliptica extracts and fractions as an antitumor agent appears to be safe, because cell viability remained higher than 90% in the cytotoxicity assay with murine fibroblast cultures (BALB/c 3T3, ATCC CCL-163), in the concentration range where the best antitumor activity (<30 &#181;g/mL) against MCF-7 and PC-3 was found.

Made available in DSpace on 2014-07-29T15:12:48Z (GMT). No. of bitstreams: 1 dissertacao gilmar quimica.pdf: 1338329 bytes, checksum: bc6520969a279db69b166b4914defad2 (MD5) Previous issue date: 2009-09-25 / This work describes the study of the specie Salacia elliptica, a plant from Hippocrateaceae family, known as bacupari, saputá and other popular names. In folk medicine it has been used in fever, swelling, scarring of wounds, diabetes and other treatments. The families Hippocrateaceae and Celastraceae are morphologically very close. Some taxonomists have considered them as only one family, while others split them in two. The Hippocrateaceae family comprises around 30 genera, and Salacia and Hippocratea are the most important with 300 species. Celastraceae comprises about 55 genera and 850 species. In this work, S. elliptica was considered an Hippocrateaceae plant, and by phytochemical study we intend to contribute to the chemotaxonomy position of this family. The phytochemical study of stem and root led to the isolation of the quinonamethide triterpenes: maitenin, pristimerin and 30-hydroxy-friedelan-3-one. Their structures were elucidated by 1H and 13C NMR spectra. The bioassays carried out so far demonstrate the great potential of this specie / Salacia elliptica é uma planta da família Hippocrateaceae, conhecida como bacupari, saputá entre outros nomes populares. Na medicina popular tem sido usado no tratamento da febre, edemas, cicatrizações de ferimentos, diabetes, etc. A família Hippocrateaceae é constituída por 30 gêneros, sendo Salacia e Hippocratea os maiores representantes. Há um embate entre grupos de botânicos quanto ao posicionamento taxonômico da S. elliptica devido às similaridades morfológicas e químicas entre as famílias Celastraceae e Hippocrateaceae. Neste trabalho, foi considerado que a S. elliptica é uma Hippocrateaceae. Trabalho este que busca contribuir com a quimiotaxonomia da família, através do isolamento de marcadores químicos. O estudo fitoquímico do caule e da raiz levou ao isolamento de triterpenos quinonametídeos: maitenina, pristimerina e 30-hidroxi-friedelan-3-ona, cujas estruturas foram elucidadas a partir de espectros de RMN de 1H e de 13C. Os bioensaios realizados até então demonstram o grande potencial da espécie

Nos travaux de doctorat s’inscrivent dans la démarche de la valorisation de la biodiversité végétale cambodgienne. Les objectifs portaient sur les travaux phytochimique et pharmacologique de deux plantes cambodgiennes : Vernonia cinerea et Vernonia elliptica. Une méthode de dosage par CLHP du composé 8α-tigloyloxyhirsutinolide 13-O-acetate (H1) appartenant à la classe des lactones sesquiterpènes de V. cinerea a été mise au point et validée dans le but de déterminer la teneur de ce composé. La teneur en H1 varie entre 0,08 % à 0,17 % pour les échantillons étudiés provenant de 4 régions du Cambodge. L’évaluation de l’activité antiproliférative in vitro des extraits de V. cinerea, et du composé H1 a été réalisée vis-à-vis de deux lignées cellulaires cancéreuses humaines HT29 et HepG2. Le composé H1 présente une CI50 similaire à celle du contrôle positif 5FU, 4,3 ± 0,2 versus 5,4 ± 0,9 µM vis-à-vis de la lignée HepG2. Les résultats intéressants obtenus sur V. cinerea nous ont incités à travailler sur d’autres espèces du genre Vernonia qui poussent au Cambodge, Vernonia elliptica. L’étude phytochimique menée sur cette plante a permis d’isoler un lignane [(+)-Syringaresinol] et deux lactones sesquiterpènes [(+)-8,13-diacetyl-piptocarphol et Glaucolide B]. Ces composés sont décrits dans cette plante pour la première fois. L’étude in vitro de l’activité antiproliférative des extraits de V. elliptica montre que l’extrait dichlorométhane obtenu à partir de l’extrait aqueux est très actif. / Our PhD work is in the process of valorization of the Cambodian plant biodiversity. The objectives focused on phytochemical and pharmacological studies of two Cambodian plants of the Vernonia genus: Vernonia cinerea and Vernonia elliptica.An analytical HPLC method was developed and validated to quantify the major compound 8α-tigloyloxyhirsutinolide 13-O-acetate (H1) of Vernonia cinerea. We found that the compound H1 content ranges from 0.08% to 0.17% in samples collected from cambodian regions. We thus evaluated the anti-proliferative activity in vitro of extracts of V. cinerea and H1 compound against two human cancer cell lines HT29 and HepG2. For HepG2 cell line, the compound H1 has a similar IC50 to the positive control 5FU, 4.3 ± 0.2 versus 5.4 ± 0.9 µM. The interesting results on V. cinerea encouraged us to study other species of Vernonia genus growing in Cambodia, Vernonia elliptica. The phytochemical study conducted on this plant led to isolate a lignan [(+)-syringaresinol] and two sesquiterpene lactones [(+)-8,13-diacetyl-piptocarphol and Glaucolide B]. These compounds were described in this plant for the first time. The in vitro study of the anti-proliferative activity of the extracts of V. elliptica shows that the dichloromethane extract obtained from aqueous extract is very active.

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No. of bitstreams: 1 TeseLPC.pdf: 5483250 bytes, checksum: fa7b00ca116fc99170337aefc31b986d (MD5) Previous issue date: 2016-03-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Brazil by presenting incomparable biodiversity, distributed in mosaic wonderfully distinct biomes becomes a promising the field of study in seeks of new phytochemicals extracted from native plants with biological activity on weeds and insects of medical importance. Species of Lauraceae and Dilleniaceae family have an immense potential in the production of secondary compounds with herbicide and insecticide properties. Thus, this study objective was to clarify the following questions: leaf extracts the stem and root Ocotea pulchella and Davilla elliptica could interfere with the growth and development of Triticum aestivum, Euphorbia heterophylla and Panicum maximum and thus make a new alternative for the control of weeds? Amongst the donor species and crude extracts which more promising to start a bioprospecting, allowing isolate and identify the present compounds? The extracts show insecticidal activity against eggs and larvae of Aedes aegypti? Coleoptile bioassays were performed using hexane extracts, dichloromethanic, acetoetilicos, methanolic and aqueous leaf, stem and root of O. pulchella and D. elliptica 0.2, 0.5 and 0.8 mg.mL-1. Most active extracts have been evaluated on the germination and early seedling growth of E. heterophylla and P.maximum using doses of 0.25, 0.5, 1 and 2 mg.mL-1. The larvicidal activity was evaluated at concentrations of 0.25, 0.5, 1, 2 and 3 mg.mL-1 and ovicidal effect with the LC50 and LC90 lethal concentrations estimated in test A. aegypti larvae. The leaf extracts O. pulchella inhibit coleoptile growth of wheat and seed germination and seedling growth of weeds and standard species. In general, the ethyl acetate extract O. pulchella showed higher phytotoxic activity. Eleven compounds were isolated and characterized. Compounds 1, 2, 4 and 6 have not been identified before in O. pulchella and Ocoteol was first described in the literature. The compounds 5, 10 and 11 were most active with superior inhibition values of 90% in the standard species. P. maximum has been the target species most affected by O. pulchella and D.elliptica leaf extracts in all evaluated parameters. The reduction in root growth of wild poinsettia seedlings can be related to the decrease in the elongation of metaxylem cells submitted at the ethyl acetate extract (O. pulchella) and methanol (D.elliptica). The extracts from root hexane and dichloromethane O. pulchella at a concentration 3 mg.mL-1 have produced bigger effect larvicide. Lower values of CL50 and CL90 were estimated for larvae subjected to the root extract from O. pulchella dichloromethane. Hatching rate, TME and VME have been affected by DCM and hexane root extracts from O. pulchella. Specie O. pulchella showed higher potential herbicide on the germination and the growth pattern and weed species and insecticide on larvae and eggs of A. aegypti. / O Brasil, por apresentar uma grande biodiversidade, torna-se um campo de estudo promissor na busca de novos fitoquímicos extraídos de plantas nativas, com atividades biológicas sobre plantas infestantes e insetos de importância médica. As espécies da família Lauraceae e Dillineaceae possuem potencial na produção de compostos secundários com propriedades herbicida e inseticida. Sendo assim, o presente estudo objetivou esclarecer os seguintes questionamentos: os extratos de folha, caule e raiz de Ocotea pulchella e Davilla elliptica interferirem no crescimento e desenvolvimento de Triticum aestivum, Euphorbia heterophylla e Panicum maximum possibilitando, assim, uma nova alternativa no controle de plantas infestantes? Entre as espécies doadoras , qual o mais promissor para se iniciar uma bioprospecção, permitindo isolar e identificar os compostos presentes? Os extratos brutos de ambas as espécies apresentam atividade inseticida sobre ovos e larvas de Aedes aegypti? Para tanto, foram realizados bioensaios de coleóptilos de trigo utilizando os extratos hexânicos, diclorometânicos, acetoetílicos, acetônicos, metanólicos e aquosos de folha, caule e raiz de O. pulchella e D. elliptica a 0,2, 0,4 e 0,8 mg.mL-1. Os extratos mais ativos foram avaliados sobre a germinação e o crescimento inicial de plântulas de E. heterophylla e P. maximum utilizando as doses de 0,25, 0,5, 1 e 2 mg.mL-1. Para os compostos puros foi utilizado as doses de 10-3, 3x10-4, 10-4, 3x10-5 e 10-5. A atividade larvicida foi avaliada nas concentrações de 0,25, 0,5, 1, 2 e 3 mg.mL-1 e o efeito ovicida por meio das concentrações letais CL50 e CL90, estimadas no teste com larvas de A. aegypti. Os extratos foliares de O. puchella inibiram o crescimento de coleóptilos de trigo, a germinação e o crescimento das plântulas das espécies daninhas e das espécies padrão. Em geral, o extrato de acetato de etila extraído de folha de O. pulchella apresentou maior atividade fitotóxica. Onze compostos do extrato de acetato de etila foram isolados e caracterizados. Os compostos dihidroxi-eudesma, hidroxi-eudesma, loliolide e ácido dimorfecolico não foram descritos antes em O. pulchella e o Ocoteol foi descrito pela primeira vez na literatura. Os compostos espatulenol, benzil salicilate e benzil benzoate foram os mais ativos, com valores de inibição superior a 90% nas espécies padrão. P. maximum foi a espécie alvo mais afetada pelos extratos de folha de O. pulchella e D.elliptica em todos os parâmetros avaliados. A redução no crescimento radicular das plântulas de amendoim-bravo pode estar relacionada com a diminuição no alongamento das células do metaxilema submetidas ao extrato de acetato de etila (O. pulchella) e metanólico (D.elliptica). Os extratos hexânico e diclorometânico de raiz de O. pulchella, na concentração de 3 mg.mL-1 produziram maior efeito larvicida. Os menores valores das CL50 e CL90 foram estimados para larvas submetidas ao extrato de raiz de O. pulchella extraído com diclorometano. A taxa de eclosão, tempo médio e velocidade média de eclosão foram afetados pelos extratos diclorometano e hexano de raiz de O. pulchella. A espécie O. pulchella apresentou maior potencial como herbicida sobre a germinação e o crescimento de espécies padrão e infestantes e atividade inseticida sobre larvas e ovos de A. aegypti.

In impoverished and resource limited communities such as townships, and welfare organizations, areas such as living and sleeping spaces are sometimes shared with animals, and occasionally humans. Dogs play an integral role in our lives and have become part of the family. Therefore, it is probable that ectoparasites, such as ticks, that feed on dogs also feed on other vertebrates, thereby, transmitting pathogens. The primary aim of this study was to screen for the presence of tick-borne pathogens in dogs from welfare organisations and townships in Cape Town, with special focus on Ehrlichia and Babesia spp. The reason for this choice of subject is due to the fact that very few tick-borne infection studies have focused on resource limited communities. Furthermore, welfare organisations have continuously attracted awareness due to the amount of unrestricted work performed by veterinarians in communities with limited resources. Consequently, the topic was borne. A total of 126 blood samples and 509 ticks (adults and nymphs) were collected directly from dogs from four welfare organisations and two townships in Cape Town. Samples were collected from April to July 2014. The four welfare organisations where samples were collected included the Animal Anti Cruelty League welfare organisations in Epping and Bellville, the Lucky Lucy Foundation in Joostenberg Vlakte and The Emma Animal Rescue Society (TEARS), located in the Sunnydale area. Samples were also collected from the Asanda village and Nomzamo, two townships located just outside the Cape Town suburb, the Strand. DNA was extracted from blood and ectoparasites and screened for the presence of Ehrlichia, Anaplasma, Theileria and Babesia species infections using touchdown PCR and RLB hybridization assays. Genus and species-specific probes were used during hybridization in order to identify specific parasite infections in the blood samples and the tick samples pooled according to geographical origin and species. Forty six (36.5%) of the blood samples tested positive for tick-borne pathogen DNA. Of the positive blood samples, 17 (13.5%) were infected with Ehrlichia canis; 16 (12.7%) with Babesia rossi and four (3.2%) samples were infected with Babesia vogeli. Incidental infections were also detected, these included Ehrlichia ruminantium (n=6, [4.7%]), Theileria taurotragi (n=2, [1.6%]) and Anaplasma sp. Omatjenne (n=1, [0.8%]) infections. DNA detected from 10 samples (7.94%) hybridized only to the Ehrlichia/Anaplasma genus-specific probes and four samples (3.17%) hybridized only to the Theileria/Babesia genus-specific probes. None of these 14 samples hybridized to any of the species-specific probes. Collected Rhipicephalus sanguineus (n=457) and Haemaphysalis elliptica (n=52) ticks were grouped into 15 pools, representing both tick species according to specific collection locations. Since only two H. elliptica from Asanda and one R. sanguineus from TEARS were collected, these ticks were mixed in pools of the dominant species as they were too few for DNA extraction. Ticks were collected from the Nomzamo Township (R. sanguineus n=400), Asanda village (H. elliptica n=2; R. sanguineus n=42), TEARS (H. elliptica n=21; R. sanguineus n=1), and the Animal Anti Cruelty League in both Epping (R. sanguineus n=14), and Bellville (H. elliptica n=29), in Cape Town. Analysis by the RLB assay showed that 11 (73.3%) of the 15 tick pools representing both tick species were positive for at least one parasite species. All positive samples hybridized with the Ehrlichia/Anaplasma genus-specific probe. Three (20%) tick pools containing both tick species tested positive for Ehrlichia canis infection, two (13.3%) tested positive for Babesia rossi and Babesia vogeli DNA was identified in one (6.6%) tick pool. The Theileria/Babesia genus-specific probe hybridised in three (20%) tick pools. These three pools were comprised of both R. sanguineus and H. elliptica tick species. These tick pools also tested positive for a specific Babesia tick-borne pathogen. Tick-borne pathogen DNA could not be detected in four (26.6%) tick pools. The fore-mentioned tick-borne pathogen DNA detected in the dog blood samples, and the ectoparasites collected from the same dogs during this study, suggests that dogs play a large role in the endemicity of these pathogens / Environmental Sciences / M. Sc. (Life Science)