Copies no. 3007841664 and 3007841665 are photocopies of the original. / Thesis (MScAgric)-- University of Stellenbosch, 2003. / ENGLISH ABSTRACT: Buchu (Agathosma betulina and A. crenulata) are grown commercially as an aromatic crop and are endemic to the Western Cape of South Africa. Poor rooting of cuttings have limited the development of superior clones. Under standard mist bed conditions terminal, sub-terminal or basal stem cuttings were taken from March to August. When not treated with an auxin, rooting percentages of between 20 and 25 were obtained. Rooting percentages increased to between 40 and 45 after treatment with 500-1000 ppm indolebutyric acid (lBA). Substituting lBA with naphthaleneacetic acid (NAA) did not improve rooting. There was a tendency for cuttings with fewer than four leaf pairs to give lower rooting percentages. Plants of Agathosma betulina x A. crenulata, grown in Paarl, and A. betulina, grown in Piketberg, were used as source plants for making cuttings. Paarl plants were shaded with 80 percent shade and Piketberg plants with 60 or 80 percent shade respectively from February to October 2002. Plants in full sun served as a control. Plants were pruned back initially in February and then two months before samples were taken in March, June, August and October at both locations. New shoots were used as cuttings. Terminal cuttings for rooting and for carbohydrate analyses were collected on four different dates (March, June, August and October). Cuttings were treated with 500 ppm indolebuteric acid (lBA) and placed in misting beds with bottom heating (18-25°C) for a period of three months. Shading reduced rooting of cuttings from the Paarl plants. However, it did not significantly increase rooting of cuttings taken from Piketberg plants. Rooting percentage was the highest in August (43%) for cuttings from sun grown plants in Paarl. No consistent relationship between, respectively, dry mass or carbohydrate content of cuttings and rooting could be established. Terminal current years' growth, taken from Agathosma crenulata x A. betulina (hybrid) softwood cuttings, collected in January 2002, were extracted with methanol and fractioned by thin layer chromatography (Silica gel) in isopropanol: acetic acid: water (4: 1:1 v/v). The chromatographs were divided in ten fractions and were bio-assayed for a rooting co-factor with the mung bean rooting test. Extracts from buchu cuttings showed significant activity at the Rf values of co-factor 3. Co-factors 1,2 and 4 do not seem to be present in significant quantities. However, co-factors with Rf values different from previous reported values were present in significant quantities. No inhibition was found in buchu. In fact, all Rf values stimulated rooting. / AFRIKAANSE OPSOMMING: Boegoe (Agathosma betulina x A. crenulata) word kommersieël verbon as 'n aromatiese gewas en is endemies tot die Wes-Kaap. Die ontwikelling van superieure klonale materiaal word beperk deur swakbeworteling. Terminale, sub-terminale en basale steggies is gesnyonder standaard misbed toestande van Maart tot Augustus. Beworteling was tussen 20 en 25 persent as geen ouksien gebruik word nie. As indolebottersuur (IBS) gebruik word tussen 500-1000 dpm, verhoog die bewortelingspersentasie tot tussen 40 en 45 persent. Die gebruik van naftaleen asynsuur (NAS) in plaas van IBS het nie beworteling verbeter nie. Daar was a tendens dat steggies wat minder as vier blaarpare gehad het 'n verlaging in bewortelingspersentasies gehad het. Plante van Paarl, A. betulina x A. crenulata, en Piketberg, A. betulina, is gebruik as plantmateriaal vir steggies. Plante in die Paarl was onder 80 persent skadu geplaas en plante in Piketberg onder 60 en 80 persent skadu van Februarie tot Oktober 2002. Plante in vol son was as 'n kontrole gebruik. Plante was eers in Februarie teruggesny en dan weer twee mande voor monsters geneem is. Die monsters is in Maart, Junie, Augustus en Oktober geneem in beide liggings. Terminale steggies is vier keer ingesamel (Maart, Junie, Augustus en Oktober) vir beworteling en koolhidraat analises. Die steggies is met 500 dpm IBS behandel. Daarna is die steggies vir drie maande in die misbed geplaas met bodem-verhitting (18- 25°C). Dit is gevind dat die gebruik van skadu die beworteling in Paarl verminder het alhoewel die beworteling in Piketberg nie beduidend beinvloed is nie. Die hoogste bewortelingspresentasies is waargeneem in Augustus (43%) in Paarl van plante wat in vol son was. Geen verband tussen onderskeidelik die droe massa of koolhidraat inhoud en beworteling kon gevind word nie. Terminale steggies van dieselfte jaar se groei van Agathosma betulina x A. crenulata (hibried) is in Januarie 2002 ingesamel. Die materiaal is geëkstraheer en gefraksioneer deur dunlaag kromatografie in isopropanol: asynsuur: water (4: 1:1 v/v). Die kromograaf is in 10 fraksies verdeel. Die fraksies was bioassaieer VIr beworteling ko-faktore met die mungboontjie bewortelingstoets. Die ekstrakte van boegoe het beduidende aktiwiteit by die Rf waardes van ko-faktor 3 getoon. Ko-faktore 1, 2 en 4 is nie in groot genoeg hoeveelhede waargeneem nie. Ko-faktore, wat nie voorheen gevind is nie, is waargeneem in beduidende hoeveelhede. Geen inhibitors is in boegoe gevind nie en al die getoetste ko-faktore het beworteling gestimuleer.

Resumo: O uso de fitoterápicos padronizados com eficácia e segurança garantidas tem mostrado resultados surpreendes em diversos países. Contudo, o Brasil ainda possui deficiências na correta caracterização dos produtos brutos e falta de um controle de qualidade mais rigoroso para esses materiais. As etapas necessárias para utilização de uma planta como fitoterápico são, primeiramente, a comprovação da sua eficácia e segurança através de ensaios farmacológicos seguidas da correta caracterização química do extrato e quantificação de alguns marcadores escolhidos para representarem a sua identidade. Astronium urundeuva é uma planta brasileira utilizada na medicina popular, para a qual são reportadas atividades antiulcerogênica e antiinflamatória, assim como efeito no trânsito gastrointestinal e em colites, sendo um alvo interessante para utilização como fitoterápico. Extratos foram preparados por percolação como descrito pela farmacopéia brasileira e avaliados por ESI-IT-MS e LC-MS apresentando em sua composição majoritária galotaninos e derivados. A caracterização química foi obtida através de duas diferentes abordagens, para os compostos majoritários análise por FIA-ESI-IT-MS forneceram indício sobre quais as substâncias presentes no extrato e HPLC-PDA com adição de padrão confirmou as identidades dessas substâncias, enquanto que os compostos minoritários tiveram que passar por uma etapa adicional de pré-concentração utilizando GPC em Sephadex LH20, depois da qual as substâncias foram identificadas ou pelo mesmo procedimento que os compostos majoritários ou por isolamento e caracterização utilizando HPLC em escala semipreparativa e espectroscopia de NMR. Dessa forma foi possível a identificação de 10 compostos fenólicos, assim como 8 galotaninos os quais foi possível... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The use of standardized phytotherapics with attested efficacy and safety has shown surprisingly good results in many countries. Nevertheless, Brazil still has deficiencies at the correct chemical characterization of raw products, and lack of a more rigorous quality control of those materials. The necessary steps for the use of a plant as a phytotherapic are, at first, proof of the efficacy and safety through pharmacological essays followed by the correct chemical characterization of the extract and quantification of some markers chosen to represent its identity. Astronium urundeuva is a Brazilian traditional herb used in folk medicine, it is reported that it shows antiulcerogenic and anti-inflammatory activity, as effect at the gastric transit and colitis, being an interesting target to be used as a phytotherapic. Extracts were prepared by percolation as described by Brazilian Farmacopeia and screened by FIA-ESI-IT-MS and LC-MS showing a major composition of gallotannins and derivatives. The chemical characterization was achieved with two different approaches, for the major compounds FIA-ESI-IT-MS analysis provided a clue about which substances were present at the extract and HPLC-PDA with standard addition confirm the identities of these substances, while the minor compounds had to go through an additional step of pre-concentration achieved by gel permeation chromatography using Sephadex LH20, after that the substances were identified either by the same procedure as for the major compounds or by isolation and characterization using HPLC in semi preparative scale and NMR spectroscopy. This way it was possible to identify 10 phenolic compounds, also 8 gallotannins which it was possible only to attribute the degree of polymerization since they are hardly isolated and there are no standards available. An HPLC-PDA... (Complete abstract click electronic access below) / Orientador: Wagner Vilegas / Banca: Jairo Kenupp Bastos / Banca: Lourdes Campaner dos Santos / Banca: Carmen Lúcia Cardoso / Mestre

Submitted by ISABELA CRISTINA GOMES HONÓRIO null (isabelagomeshonorio@gmail.com) on 2016-05-25T19:12:00Z No. of bitstreams: 1 ESTUDO DA DIVERSIDADE GENÉTICA E QUÍMICA DE Uncaria tomentosa Willd. ex Roem. & Schult. E Uncaria guianensis Gmell DE POPULAÇÕES NATURAIS LOCALIZADAS NA AMAZÔNIA..pdf: 2336644 bytes, checksum: 05f34f338ce3e8cd544dea281a52873d (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-05-30T17:46:08Z (GMT) No. of bitstreams: 1 honorio_icg_dr_bot.pdf: 2336644 bytes, checksum: 05f34f338ce3e8cd544dea281a52873d (MD5) / Made available in DSpace on 2016-05-30T17:46:08Z (GMT). No. of bitstreams: 1 honorio_icg_dr_bot.pdf: 2336644 bytes, checksum: 05f34f338ce3e8cd544dea281a52873d (MD5) Previous issue date: 2016-03-11 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Uncaria tomentosa Willd. ex Roem. & Schult. e Uncaria guianensis Gmell. são plantas medicinais nativas da Amazônia, utilizadas na medicina tradicional com ação anti-inflamatória, sofrem pressão antrópica como o desmatamento e extrativismo predatório para uso e necessitam de cuidados em relação à conservação. U. tomentosa é uma das plantas medicinais que compõe a lista da relação nacional de medicamentos essenciais (RENAME) disponibilizada pelo Ministério da Saúde para os municípios brasileiros, através do Sistema Único de Saúde (SUS) e a U. guianensis apesar de não estar nesta lista apresenta também alcaloides oxindólicos pentacíclicos, como marcadores químicos, sendo utilizada popularmente como anti-inflamatória e sua ocorrência na natureza é maior quando comparada à U. tomentosa. Os objetivos desse trabalho foram avaliar a diversidade genética e química entre os indivíduos de U. tomentosa e U. guianensis por marcador molecular SRAP (Sequence-Related Amplifield Polymosphism) e quantificar os alcaloides oxindólicos pentacíclicos mitrafilina e isomitrafilina em folhas por HPLC. A coleta do material foi feita nos estados do Acre, Amazonas, Amapá e Pará. Para ambas as espécies foram coletadas oito populações, em municípios distintos com 20 indivíduos cada. A genotipagem de U. tomentosa foi realizada utilizando três combinações de primers e os fragmentos submetidos à eletroforese em gel de poliacrilamida. A genotipagem de U. guianensis foi realizada usando quatro combinações de primers e os fragmentos analisados no equipamento 4300 DNA Analyser LI-COR®. A quantificação dos alcaloides oxindólicos pentacíclicos mitrafilina e isomitrafilina foi realizada de acordo com método já descrito na literatura com algumas modificações. Nas populações avaliadas de U. tomentosa a variabilidade genética foi maior dentro (75%) das populações do que entre elas (25%). O valor de Fst foi de 0,246, evidenciando que as populações estão estruturadas em Modelo de Ilhas. A maior porcentagem de locus polimórficos (95,68%) e a maior diversidade genética foram encontradas na população coletada no município de Mâncio Lima - AC. O fluxo gênico foi considerado restrito (Nm=1,57) e não houve correlação entre as distâncias geográfica e genética. Para a espécie U. guianensis, a variabilidade genética também foi maior dentro das populações (81%) do que entre elas (19%). O valor de Fst foi de 0,188, o que mostra que as populações estão se aproximando do Modelo de Ilhas. A maior porcentagem de locus polimórficos (90,21%) e a maior variabilidade genética foram verificadas na população coletada no município de Mazagão-AC. O fluxo gênico foi considerado baixo (Nm = 2,57) e houve fraca correlação entre as distâncias geográfica e genética, portanto considerada não significativa. A maior concentração dos alcaloides mitrafilina (11,17 mg.g-1.PS) e isomitrafilina (2,99 mg.g-1.PS) em U. tomentosa foram encontradas nos indivíduos da população de Tarauacá-AC e em U. guianensis foi de 1,09 mg.g-1.PS de mitrafilina e 0,29 mg.g-1.PS de isomitrafilina encontradas na população de Boca do Acre-AM. Além disso, para essa espécie, foi possível quantificar mitrafilina em apenas 24,8% dos indivíduos e a isomitrafilina em 20,4% dos indivíduos estudados. Conclui-se que a conservação de ambas as espécies deverá ser realizada com a coleta de um maior número de indivíduos nas populações com maiores variabilidades genética e química. / Uncaria tomentosa Willd. ex Roem. & Schult. and Uncaria guianensis Gmell. are medicinal plants natives from Amazonia state, used in the ocidental medicine for its anti-inflammatory effect, which are suffering from the actions of mankind by deforestation and predatory extraction, requesting care for their conservation. U. tomentosa is one of the medicinal plants that make up the national ratio of the list of essential drugs (RENAME) made available by the Ministry of Health to the municipalities through the Unified Health System (SUS) and U. guianensis although this is not list also features pentacyclic oxindole alkaloids, as markers, being popularly used as anti-inflammatory and its occurrence in nature is higher compared to U. tomentosa. The objectives of the present study were to evaluate the chemical and genetical diversity between individuals of U. tomentosa and U. guianensis by molecular marker SRAP (Sequence-Related Amplifield Polymosphism) and quantify pentacyclic oxindolic alkaloides mitraphylline and isomitraphylline in leaves via HPLC. Samples were collected in the states of Acre, Amazonas, Amapá and Pará. For both species, 20 samples were collected from each of the eight populations coming from different locations. The genotyping of U. tomentosa was realized by using the combination of three primers and the fractions submitted to an electrophoresis in acrylamide gel. The genotyping of U. guianensis was realized by using the combination of four primers and the fractions analyzed with a 4300 DNA Analyser LICOR®. The quantification of pentacyclic oxindole alkaloides mitraphylline and isomitraphylline were realized by using the existing method with some modifications. The U. tomentosa populations showed higher genetic variability inside the same population(75%) than between different populations (25%). The value of Fst was 0,246, showing that the populations are following the Island Model. The highest percentage of polymorphic locus (95,68%) and the highest genetic diversity were found in the population from Mâncio Lima - AC. The gene traffic was considered restrictive (Nm=1,57) and there was no correlation between geographic and genetic distances. For U. guianensis, the genetic variability was higher inside same populations (81%) than between different populations (19%). The value of Fst was 0,188, showing that the populations are coming close to the Island Model (modelo de ilhas). A higher percentage of polymorphic locus (90,21%) and a higher genetic variability were confirmed in the population colected in Mazagão - AC. The gene traffic was considered low (Nm = 2,57) and as having a weak correlation between geographic and genetic distances and, therefore, considered not significant. The highest alkaloid concentrations of mitraphylline (11,17 mg.g-1 .PS) and isomitraphylline (2,99 mg.g-1 .PS) in U. tomentosa were found in samples of the population from Tarauacá - AC and in U. guianensis samples showed alkaloid concentrations of 1,09 mg.g-1 .PS of Mitraphylline and 0,29 mg.g-1 .PS of Isomitraphylline in the population from Boca do Acre - AM. Besides that, to this species, it was possible to quantify mitraphylline in only 24.8% of individuals isomitraphylline and 20.4% of subjects studied. It’s concluded that the conservation of both species should be done with the collection of a greater number of individuals in populations with higher genetic and chemical variability.

Made available in DSpace on 2015-05-14T12:59:31Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1623079 bytes, checksum: 1db342a9defb06c8f56de918c6d74da7 (MD5) Previous issue date: 2012-02-13 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Mimosa tenuiflora (Willd.) Poir., popularly known as Jurema preta, is a plant family Mimosaceae, found mainly in the states of Piauí, Ceará, Rio Grande do Norte, Paraiba, Pernambuco, Alagoas, Sergipe and Bahia, which this species is typical of semi-arid areas of Brazil. In folk medicine, the bark of the stem is the main part of the plant used to treat various ailments such as burns and inflammations. This study aimed to investigate the cytotoxicity and genotoxicity of crude ethanolic extract (BSE) and Mimosa tenuiflora in prokaryotic and eukaryotic cells in order to encourage later, safely, its use as a source of potentially therapeutic drugs or act as pharmacological tools. To this end, we evaluated the antimicrobial activity against Gram negative and Gram positive clinical importance, investigated the potential cytotoxic utilizing the hemolytic activity and osmotic fragility (anti-hemolytic), as well as potential antioxidant and anti-oxidant in human erythrocytes, there was observed the potential mutagenic and anti-mutagenic in bacterial cells through the Ames test and also evaluated the clastogenic and aneugenic potential using the micronucleus test in peripheral blood of mice in vivo. The results showed that the EEB and the tannins of M.tenuiflora had antibacterial activity against all strains of Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli tested and this was characterized as strong with MIC ranging from 62,5 to 250 μg.mL-1. In the investigation of cytotoxic of M.tenuiflora it was observed that the extract did not induce a significant degree of hemolysis and even protected the erythrocyte membrane against hemolysis. In tests of the oxidizing activity of M.tenuiflora, this plant did not cause oxidation of hemoglobin and tests of the extract antioxidant activity behaved as a powerful antioxidant. In the investigation of genotoxicity, we observed that M.tenuiflora was not mutagenic, or in vitro tests and in vivo. Thus, the extract of M.tenuiflora has no cytotoxic or genotoxic that compromise its use as a source of potentially therapeutic drugs. / Mimosa tenuiflora (Willd.) Poir., popularmente conhecida como jurema preta, é uma planta da familia Mimosaceae, encontrada principalmente nos estados do Piauí, Ceará, Rio Grande do Norte, Paraíba, Pernambuco, Alagoas, Sergipe e Bahia, sendo esta espécie típica das áreas semi-áridas do Brasil. Na medicina popular, a casca do caule é a principal parte da planta utilizada no tratamento de diversas enfermidades como infecções. O presente trabalho teve como objetivo investigar a citotoxicidade e a genotoxicidade do extrato etanólico bruto (EEB) da Mimosa tenuiflora em células procarióticas e eucarióticas com a finalidade posterior de incentivar, com segurança, a sua utilização como fonte de drogas potencialmente terapêuticas ou que atuem como ferramentas farmacológicas. Para tanto, avaliou-se a atividade antimicrobiana frente a bactérias Gram negativas e Gram positivas de importância clínica, investigou-se o potencial citotóxico utilizando como parâmetro a atividade hemolítica e a fragilidade osmótica (anti-hemolítica), assim como o potencial oxidante e antioxidante em eritrócitos humanos, observou-se o potencial mutagênico e antimutagênico em células bacterianas através do teste de Ames e também avaliou-se o potencial clastogênico e aneugênico através do teste do micronúcleo em sangue periférico de camundongos in vivo. Os resultados mostraram que o EEB e os taninos de M.tenuiflora tiveram atividade antibacteriana contra toda as linhagens de Staphylococcus aureus, Pseudomonas aeruginosa e Escherichia coli testadas, e esta foi caracterizada como forte com CIM variando de 62,5 a 250 μg.mL-1. Na investigação citotóxica do EEB de M.tenuiflora observamos que o extrato não induziu um grau significativo de hemólise e ainda protegeu a membrana do eritrócito contra hemólise. Nos testes de atividade oxidante o EEB de M.tenuiflora não provocou oxidação da hemoglobina e nos testes de atividade antioxidante o extrato se comportou como um poderoso agente antioxidante. Já na investigação genotóxica, observamos que o EEB da M.tenuiflora não foi mutagênico, nem nos testes in vitro nem in vivo e apresentou efeito antimutagênico em algumas linhagens ensaiadas. Sendo assim, a baixa atividade citotoxicidade e genotoxicidade de M.tenuiflora não comprometem seu uso como fonte de droga potencialmente terapêutica.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A utilização de ervas e plantas para finalidades terapêuticas é bastante antiga e registrada em várias sociedades. Esta pesquisa procura investigar como se dá o processo de construção e reprodução do conhecimento em medicina popular (particularmente o cuidado com ervas e plantas medicinais) através da história de vida de terapeutas populares pessoas que se tornaram referências em suas comunidades por adquirir, praticar e transmitir esse conhecimento. Tem como objetivos conhecer e compreender o processo de construção do conhecimento sobre plantas medicinais por terapeutas populares do município do Rio de Janeiro; compreender as formas de reprodução deste conhecimento; entender o contexto de relações destes terapeutas com o saber científico. Utilizou-se como opção metodológica a história oral com ênfase na história de vida. Foram entrevistados sete terapeutas populares de diferentes bairros do município. Observou-se que o conhecimento em relação às ervas medicinais não está restrito a um pequeno grupo e limitado a uma forma de transmissão oral. O conhecimento tem se reproduzido, mas também tem se modificado, mesclando-se com outras formas de conhecimento e entremeado por diversas fontes de informação que vão desde a transmissão oral até informações disponíveis na mídia. A principal forma de reprodução do conhecimento parece ser a perpetuação do negócio, que evoca o discurso do tradicional, a natureza das ervas e suas propriedades benéficas e se sustenta por uma rede de informação que se utiliza dos diversos meios de comunicação e uma rede de produção e comércio que garante o sustento econômico dos terapeutas populares. A transformação das práticas de utilização de ervas medicinais em patrimônio cultural ou negócio de família além de garantir a perpetuação do conhecimento, garante um espaço diferente dentro das práticas de cuidado em saúde. As relações dos terapeutas com a medicina oficial não são relações de tensões, embora haja criticas, mas de interposição de sistemas terapêuticos. / The use of herbs and plants for therapeutic purposes is quite old and registered in various societies. This research seeks to investigate how the process of construction and reproduction of knowledge in folk medicine (particularly the use of herbs and medicinal plants) through the life story of popular therapists - people who have become references in their communities for acquiring, practicing and transmitting this knowledge. It aims to identify and understand the process of knowledge construction about medicinal plants by popular therapists in the city of Rio de Janeiro; to understand the forms of reproduction of this knowledge; and to understand the context of the relationship between these practitioners and the scientific knowledge. The oral history, with an emphasis on life history, was used as a methodological approach. Seven popular therapists were interviewed in different districts of the city. It was observed that the knowledge about the medicinal herbs is not restricted to a small group and limited to a form of oral transmission. Knowledge has been reproduced, but also has been modified, mingling with other forms of knowledge and punctuated by several information sources ranging from oral transmission to information available in the media. The main form of knowledge reproduction seems to be the perpetuation of a business, which evokes the traditional speech, the nature of herbs and their beneficial properties and is supported by an information network that uses various means of communication and a production and trade network that ensures economic sustenance for the popular therapists. The transformation of the practice of using herbs in cultural heritage or family business ensures the perpetuation of knowledge, and it also ensures a different space within the practices of health care. The relationship of therapists with official medicine is not a tensioned one, although there are criticisms, but one with the interposal of the therapeutic systems.

A fitoterapia e o uso de plantas medicinais fazem parte da prática da medicina popular, que diminuiu frente ao processo de industrialização nas décadas de 1940 e 1950, aumentando a busca, pela população, por terapias menos agressivas destinadas ao atendimento primário à saúde. Essa complementa o tratamento usualmente empregado para a população de menor renda. O objetivo deste trabalho foi analisar o conhecimento dos gestores e profissionais de saúde que atuam na atenção primária, sobre fitoterapia e uso de plantas medicinais, nos municípios de Cascavel e Foz do Iguaçu - Paraná. Para tanto, no período de fevereiro a julho de 2009, foi realizado estudo exploratório, descritivo, qualitativo, empregando como instrumentos de pesquisa entrevistas semiestruturadas e um questionário estruturado, auto aplicado, com perguntas fechadas, a fim de contemplar os seguintes aspectos: perfil dos profissionais; aceitabilidade quanto à implantação da fitoterapia na atenção básica; confiança nos tratamentos com fitoterápicos; capacitação e formação profissional relacionadas à fitoterapia, entre outros itens. Um dos gestores considerou recente o programa de fitoterapia. Indicadores de satisfação da população quanto ao uso dos fitoterápicos ou plantas medicinais ainda não existem. O gestor do município de Cascavel relatou interesse na implantação do programa. Os demais profissionais entrevistados, de maneira geral, relataram não haver recebido formação sobre o tema durante a graduação, nem mesmo dentro das UBS onde trabalham. Entre os profissionais entrevistados seis (60%) relataram o acesso às informações sobre fitoterapia através do conhecimento popular, uma (10%) formação na UBS, dois (20%) através de periódicos, quatro (40%) através de meio de comunicação, sendo que quatro citaram mais que uma das opções. Em Foz do Iguaçu, nas UBS onde a terapêutica está implantada, os profissionais relataram não terem sido consultados antes de sua implantação, e a avaliação das plantas utilizadas pelos pacientes atendidos ocorre somente através de avaliação objetiva e subjetiva do tratamento Para implantação da fitoterapia nas UBS desses dois municípios, é necessário capacitar os profissionais quanto ao tema, desde o cultivo até a prescrição, melhorando o uso racional desses medicamentos. / The phytoterapy and the use of medicinal plants are part of practice of the popular medicine, that decreased with industrialization, in the 1940 and 1950s, increasing the search, of population, of therapies less aggressive, in the primary health care. This practice is complementary to the usual treatment used for the poor population. The purpose of this work was analyze the knowledge of the managers and health professionals, which works in the primary attention, about phytoterapy and use of medicinal plants, in the cities of Cascavel and Foz do Iguaçu - Paraná. For this, in the period of February to July of 2009, was done an exploratory, descriptive, and qualitative study, using as research instruments halfstructuralized interviews and a structuralized, auto-applied questionnaire, with closed questions, in order to contemplate the following aspects: profile of the professionals; acceptability of the introduction of the phytoterapy in the basic attention; the trust in the treatments with phytoterapy; related qualification and professional formation to the phytoterapy, among others itens. One of the health managers judged the program of phytoterapy recent. Pointers of satisfaction of the population about the use of the phytoterapy or medicinal plants dont exist. The health manager of Cascavel related interest in the introduction of the program. The other professionals interviewed, related that didnt have received formation about the subject during the graduation, not even inside of the health basic unit where they work. Six (60%) related the access to the information on phytoterapy through the popular knowledge, one (10%) formation in the health basic unit , two (20%) through periodic, four (40%) through media, four (40%) had cited more than one of the options. At Foz do Iguaçu, in the health basic unit where the therapy is implanted the professionals had told that they arent consulted previously its implantation.The evaluation of the plants used for the patients in this city only occurs through an objective and subjective evaluation of the treatment. For implantation of the phytotherapy in the UBS of these two cities, is necessary qualification of the professionals about the subject, about the cultivation and prescription of the plants, improving the rational use of these medicines.

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No. of bitstreams: 1 domingues_n_dr_sjc.pdf: 3393326 bytes, checksum: 5504238a789ee1c43fe177b59d548064 (MD5) Previous issue date: 2018-02-07 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Os micro-organismos estão cada vez mais resistentes aos medicamentos disponíveis tanto na medicina quanto na odontologia, e esta resistência é ainda maior quando estão organizados em biofilmes mono ou multiespécies, de modo que o estudo de antimicrobianos alternativos, como fitoterápicos, estão em crescente ascensão. A interação entre leveduras e bactérias está intimamente presente na cavidade bucal, em que nichos como dentes, língua, mucosa e bolsa periodontal, nutrientes e temperatura adequados promovem condições favoráveis para formação do biofilme. Com isso, o objetivo deste trabalho foi avaliar a atividade antimicrobiana dos extratos de Pfaffia paniculata K (pfaffia), Hamamelis virginiana L. (hamamelis), Stryphnodendron barbatiman (barbatimão) e Gymnema sylvestre (gimena) em biofilmes heterotípicos de Candida albicans (ATCC 18804) com Streptococcus mutans (ATCC 35688), Staphylococcus aureus (ATCC 6538), Enterococcus faecalis (ATCC 4083) e Pseudomonas aeruginosa (ATCC 15442). Para isso, suspensões padronizadas em 107 cels/mL dos micro-organismos testes, foram distribuídos em placas de microtitulação de 96 poços, juntamente 100 µL de caldo BHI. As placas foram incubadas em estufa bacteriológica a 37ºC/48h (5% de CO2 para S. mutans) e, após, os biofilmes foram submetidos ao tratamento com os extratos por 5 min e 24 h, nas respectivas concentrações de 100 mg/mL, 50 mg/mL e 25 mg/mL. Foi utilizado solução salina 0,9% (5 min) ou caldo BHI (24 h) nos grupos controles. Após, os biofilmes foram lavados e desagregados do fundo da placa e diluições seriadas foram semeadas em ágar seletivo para cada micro-organismo. Foram realizadas contagens de UFC/mL (log10) após 24 h de incubação e analisadas estatisticamente pelo método Kruskal-Wallis suplementado pelo teste de Dun’s (p<0.05). Os resultados obtidos indicaram reduções significativas promovidas pelos extratos nos dois tempos de tratamento analisados. Foi observado que o extrato de H. virginiana apresentou redução de todos os grupos analisados no tempo de tratamento de 24 h. Conclui-se que os extratos de P. paniculata, H. virginiana, S. barbatiman e G. sylvestre apresentaram ação antimicrobiana sobre biofilmes muliespécie de C. albicans com as bactérias de interesse médico-odontológico, nos tempos de tratamento de 5 min e 24 h. / Microorganisms are increasingly resistant to drugs available in both medicine and dentistry, and this resistance is even greater when they are organized into mono or multispecies biofilms, so that the study of alternative antimicrobials, such as herbal medicines, are on the rise. The interaction between yeasts and bacteria is intimately present in the oral cavity, in which niches such as teeth, tongue, mucosa and periodontal pocket, food and adequate temperature promote adequate conditions for biofilm formation. The aim of this study was to evaluate the antimicrobial activity of extracts of Pfaffia paniculata K., Hamamelis virginiana L., Gymnema sylvestre and Stryphnodendron barbatiman M. in heterotopic biofilms of Candida albicans (ATCC 18804) with Streptococcus mutans (ATCC 35688). Staphylococcus aureus (ATCC 6538), Enterococcus faecalis (ATCC 4083), and Pseudomonas aeruginosa (ATCC 15442). Standardized suspensions at 107 cells/mL of the test microorganisms were distributed in 96 well microtiter plates together with 100 μL of BHI broth, the plates were incubated in a bacteriological oven at 37°C/48h (5% CO2 for S. mutans). After the incubation time, treatments were performed at 5 min and 24 h times, applying the respective extracts at the concentrations of 100 mg, 50 mg and 25 mg/mL and applying 0.9% saline solution or BHI broth in the control groups. After the biofilms were washed and disaggregated from the bottom of the plate, performing serial dilutions for later seeding in selective agar. UFC/ml (log10) counts were performed after 24 h of incubation and statistically analyzed Kruskal-Wallis method supplemented by the Dun's test (p<0.05%). The obtained results indicated significant reductions promoted by extracts in the two treatment times analyzed. It was observed that the extract of H. virginiana showed reduction of all the analyzed groups without treatment time of 24 h. It is concluded that the extracts of P. paniculata, H. virginiana, S. barbatiman and G. sylvestre presented antimicrobial action after analysis of the heterotypic biofilms of C. albicans with the bacteria of medical and dental interest, in the treatment times of 5 min and 24 h.

Background: Natural products have attracted high attention due to their great contribution in drug discovery. Many natural products have shown to be effectively against different disease targets including cancer, malaria and HIV. And their structural diversity is a rich resource for the discovery of novel bioactive compounds. This thesis is to explore plant natural products for their potential in treatment of malaria and cancer diseases. Malaria is still considered as a major global health problem, affecting a large population of the world, especially in the developing countries. Effective drug discovery is still one of the main efforts to control malaria, and plant-derived compounds have played the most important role for treatment of malarial disease. In our previous work, we have evaluated more than 2,000 plant extracts against the malaria parasite Plasmodium falciparum. As a result, we discovered dozens of antiplasmodium plant leads. Bioassay guided separation of these active plant extracts led to isolation of some potent antimalarial compounds. Among them, trichothecenes, the sesquiterpenes identified from the plant Ficus fistulosa and Rhaphidophora decursiva, were found to have potent inhibitory activity against P. falciparum with IC50 values in the sub-nano molar range in our previous study (Zhang et al. 2002). However, these compounds are significant cytotoxic. In order to improve the antimalarial activity of the trichothecenes, we evaluated the antimalarial activity of dozens of trichothecenes, and based on the structure-activity relationships (SAR) analysis, we synthesized trichothecene derivatives with low cytotoxicity. On the other hand, cancer has stricken one-third of the world's population. Through our anticancer drug program to discover bioactive leads from thousands of the plant extracts, the extract of Miliusa sinensis Finet et Gagnep. (Annonaceae) was found to exhibit cytotoxic activity against a panel of cancer cell lines. Our previous bioassay-directed fractionation of the leaf, twig and flower extracts of M. sinensis has led to the discovery of a novel class of anticancer lead molecules, which we designated as miliusanes (Zhang et al, 2006). M. balansae, in the same family with M. sinensis, also contains bioactive miliusanes. We thus collected the plant materials of M. balansae to isolated additional new anticancer miliusanes. Objective: The objective of this study is to discover novel antimalarial and anticancer natural compounds from plants using different techniques in combination of extensive literature review, phytochemical separation, SAR analysis, semi-synthesis and biological activity study. Trichothecenes and miliusanes are the two major classes of the compounds, which have been extensively explored in the current thesis for their antimalarial and anticancer potential.;Methodology: There was a prior comprehensive review article entitled "Antimalarial activity of plant metabolites" by Schwikkard and Van Heerden (2002), which reported structures of those antiplasmodial active compounds and covered literatures up to the year 2000. As a continuation of their work, antimalarial compounds isolated from plants, including marine plants, which reported in the literatures from 2001 to the end of 2017 have been reviewed and organized according to their plant families. Dozens of trichothecenes have been obtained by us and explored for their SARs. Based on the SAR analysis, we designed and carried out the structure modifications of some trichothecenes. These compounds were evaluated for their antimalarial and cytotoxic activities. M. balansae, in the same genus with M. sinensis, was selected to isolate bioactive miliusanes, as well as the other active components through bioassay-guided fractionation study. Columns chromatography was used in fractionation and separation of the dichloromethane extract of M. balansae. Preparative HPLC separation and LC-MS analysis were used to speed up the isolation process. All isolates were determined for their chemical structures by spectroscopic means such as NMR and MS, and then evaluated for their anticancer potential. The isolated compounds with abundant amounts were further explored to modify their structures in order to improve their biological activities. Result: During the span of the last 17 years (2001-2017), 175 antiplasmodial compounds were discovered from plants. These active compounds were organized in our review article according to their plant families. In addition, we also included ethnobotanical information of the antimalarial plants. In order to decrease toxicity while retaining antimalarial activity of the trichothecenes, we analyzed SAR of 28 trichothecene analogues. Based on the SAR analysis, we were able to conclude that the diacetylation of C-4 and C-15, and the hydroxylation at C-3 or C-4 could significantly improve the therapeutic indices of trichothecenes. Subsequently, our synthesis of a trichothecene derivative demonstrated potent inhibitory activity against P. falciparum with an IC50 value of 10.4 nM and low toxicity against KB cells with an IC50 value of 556 nM. The separation of the dichloromethane extract of M. balansae led to the isolation of 16 new miliusanes, along with 12 known ones, and 6 flavonoids. Seven of them exhibited significant cytotoxicity against human colorectal cancer cell lines HCT116 with IC50 values in the range of 1.24 - 4.2 µM. The SAR study of the miliusanes indicated that the α,β-unsaturated ketone was the active group of but may not be the essential group responsible for the bioactivity of miliusanes. Two flavonoids that showed moderate cytotoxic activity were carried out their structure activities modification. A total of 19 derivatives have been synthesized based on the two flavonoid structures.

Black Cohosh (Actaea racemosa) is one of the highest selling medicinal plants, ranking as the sixth best seller in the US in 2015 (Smith et al., 2016). However, this popularity has been tarnished by claims of hepatotoxicity. The investigation of these reports has determined that implicated products did not contain Black Cohosh plant material. Other reports were shown to be incomplete or had other factors contributing. This has led to the suspicion that cases of adverse reactions may in fact be linked to cases of substitution or adulterations with Asian species of Actaea, rather than to A. racemosa. (Jordan et al., 2010). This shows the need for authentication of Black Cohosh products. In this study various DNA based authentication methods were developed. The first, PlantID is capable of discriminating between Actaea racemosa and four potential adulterant species; Actaea cimicifuga, Actaea cordifolia, Actaea podocarpa and Caulophyllum thalictroides, in a single PCR reaction. The resulting fragments are scrutinized using gel electrophoresis. Other platforms of analysis were trialled with little success. The second was a qPCR based method. These assays are competent in detecting A. racemosa, A. cimicifuga and A. dahurica species and are compared to a generic primer capable of amplification of ten Actaea species. This enables the user to detect specific species in comparison to how much Actaea species are present as a whole. This assay was extensively tested on many materials and products available in the UK and the USA. Out of 34 products assessed it was possible to extract DNA from 32. From the UK market it was found that five products contained undeclared species. From the US market it was found that six products contained undeclared species. All of the THR registered products were found to contain only the authentic species Actaea racemosa. This was a reassuring result from the analysis and adds further value to the scheme of THR. Sequence data from GenBank was used to assist in assigning species to sequenced DNA samples. The data contained on GenBank was scrutinised using various bioinformatics tools. Sequences were organised into molecular taxonomic units using tree diagram software. This showed efficiently and iii visually which sequence entries were reliable to use based upon grouping. This analysis showed that the nuclear internal transcribed spacer (nrITS) was an ideal barcoding region and that maturase K (MatK) was a poor choice for Actaea species. To address the issue of hepatotoxicity claims, cultured human hepatocyte derived cells were treated with 60% ethanol extracts of Actaea racemosa and Asian Actaea. A qPCR array was utilised to assess 84 genes associated with hepatotoxicity across various concentrations of extract. The collective array output gave a plethora of data which was analysed using bespoke online software from the manufacturer. Stringent quality controls were included on the arrays which gave confidence of results. There were small changes noted for Actaea racemosa and some activity for the Asian Actaea treated cells was also seen. An LDH and MTT assay were used to assess cell viability and toxicity in two human hepatocyte derived cell lines. Actaea racemosa showed no significant effects whereas the Asian Actaea extract showed a notable decrease in cell viability and significant release of LDH indicating toxicity. The Asian Actaea material used to manufacture extracts was of questionable species origin but determined to be either A. dahurica or A. cimicifuga. The results from these experiments were unfortunately not as conclusive as hoped, but did show some evidence of a more likely culprit of toxicity originating from Asian Actaea species.

Orientador: Leonardo Theodoro Büll / Banca: Lin Chau Ming / Banca: Carmem Silvia F. Boaro / Banca: Antonio Francisco J. Bellote / Banca: Hellton Damin da Silva / Resumo: A espécie medicinal Maytenus ilicifolia, localiza-se naturalmente sob diferentes condições edafo-climáticas. Este fato implica em prováveis mecanismos de adaptação das populações naturais, cujas características químicas podem, ou não, estar fixadas geneticamente. A identificação da variabilidade fenotípica da espécie permite viabilizar práticas de manejo visando a produção de fitoterápicos com qualidade organoléptica, nutricional e terapêutica. Para identificar a influência do ambiente na composição fitoquímica de M. ilicifolia foram efetuados três estudos, o primeiro em quatro populações naturais distribuídas no Estado do Paraná, o segundo em um banco de germoplasma, e o terceiro em uma unidade de cultivo experimental onde foi conduzido um ensaio de fertilização baseado na aplicação de diferentes doses de um resíduo de siderurgia rico em Si. Foram avaliados os teores foliares de N, P, K, Ca, Mg, Fe, Mn, Cu, Zn, Si, Al, lignina, fenóis totais, fenóis não tanantes e taninos. Nas populações naturais também foi determinada a massa específica foliar. No banco de germoplasma, em função da elevada diversidade morfológica, foi efetuada a classificação dos indivíduos em morfotipos, tendo sido identificados nove morfotipos foliares, os quais apresentaram relação com alguns dos ambientes de origem das populações. Os estudos efetuados indicam que é possível efetuar o controle de compostos fenólicos presentes em M. ilicifolia, seja através do manejo de luz, interferindo tanto na síntese de lignina quanto dos taninos, seja por meio da aplicação de Si e Ca, regulando principalmente a síntese de fenóis totais e taninos. / Abstract: Maytenus ilicifolia is a medicinal species that occurs naturally under different soils and light conditions. This fact involves adaptation mechanisms of the natural populations whose chemical characteristics can be or not genetically fixed. The identification of the genetic variability can viabilize the management practices to the production of phytotherapics with organoleptical, nutritional and therapeuthic quality. Three studies were developed to determine the influence of the environment in the phytochemical content of M. Ilicifolia and the samples were collected from four natural communities, a germoplasm bank, and an experimental area in wich there was fertilized with different doses a rich Si residue. There was determinate the specific leaf mass in leaves from the natural populations, also there was determinate the total content of N,P, K, Ca, Mg, Fe, Mn, Cu, Zn, Si, Al, lignin, total phenols and tannins in all samples. In the germoplasm bank the individual plants were classified in nine morphotypes, too. The studies indicate that is possible to controle the M. ilicifolia lignin and tannin composition by light management, as well as the total phenols and tannin by Si and Ca fertilization. / Doutor