As raízes de Pfaffia paniculata (Ginseng brasileiro) são comercialmente encontradas como cápsulas contendo as raízes pulverizadas, misturadas ou não ao extrato etanólico destas raízes. Estas raízes são popularmente recomendadas para vários propósitos, e têm sido utilizadas na terapia contra o câncer pela medicina popular. Os principais componentes encontrados nestas raízes já isolados incluem o ácido pfáffico e os pfaffosideos A, C, D, E e F; estes componentes inibiram o crescimento de células do melanoma B-16, demonstrado em um outro estudo. O objetivo deste estudo foi investigar os efeitos dos extratos etanólico, butanólico ou aquoso das raízes de Pfaffia paniculata sobre o crescimento das linhagens tumorais de células mamárias humanas MCF-7 e SKBR-3, utilizando método colorimétrico (cristal violeta) e quantificação das células que incorporaram bromodeoxiuridina (BrdU). A coloração por laranja de acridina/brometo de etídeo foi utilizada para avaliar morte celular, e as alterações subcelulares foram avaliadas por microscopia eletrônica. O extrato butanolico apresentou efeitos citotóxicos nas linhagens MCF-7 e SKBR-3. Morte celular foi observada pelo tratamento com o extrato butanólico por 1 h; alterações morfológicas foram observadas com 500µg/mL deste extrato, após 24 h de tratamento. Após o tratamento por 48 h com o extrato butanólico nesta mesma concentração, foi observado degeneração de componentes citoplasmáticos e alterações morfológicas sugestivas de morte celular. O tratamento com 1000µg/mL deste extrato levou a profundas alterações citoplasmáticas e nucleares, incompatíveis com a viabilidade celular. O tratamento com o extrato etanólico não causou efeitos significantes no crescimento das células MCF-7 e SKBR-3; o extrato aquoso, por outro lado, estimulou o crescimento das células MCF-7, após o tratamento por 1 h. Estes resultados indicam efeitos citotóxicos exercidos pelo extrato butanólico das raízes de Pfaffia paniculata em linhagens celulares de tumor de mama humano / The roots of Pfaffia paniculata (Brazilian ginseng) are commercially available as capsules containing the powdered roots, mixed or not with the ethanolic extract of the roots. These roots have been populary recommended for many purposes, and have also been used on cancer therapy by folk medicine. The main components found in these roots that have already been isolated include pfaffic acid and pfaffosides A, C, D, E and F; these components inhibited the growth of melanoma B-16 cells, as shown in another study. The aim of this study was to investigate the effects of ethanolic, butanolic or aqueous extract of the roots of Pfaffia paniculata on the growth of human breast tumor cell lines MCF-7 and SKBR-3, using a colorimetric method (crystal violet) and quantification of bromodeoxiuridine (BrdU) positive cells. Acridine orange/ethidium bromide staining was utilized to evaluate cell death, and subcellular alteration was evaluated by electron microscopy. The butanolic extract presented cytotoxic effects in MCF-7 and SKBR-3 cell lines. Cell death was observed following treatment with the butanolic extract for 1 h; morphologic alterations were observed with 500µg/mL of this extract, after 24 and 48 h of treatment. Following treatment for 48 h with the butanolic extract with this same concentration, degeneration of cytoplasmic components and morphological alterations suggestive of cell death were observed. Treatment with 1000µg/mL of this extract lead to profound cytoplasmic and nuclear alterations, incompatible with cell viability. The treatment with ethanolic extract didn´t lead to significant effects on the growth of MCF-7 and SKBR-3 cells; the aqueous extract, on the other hand, stimulated the growth of MCF-7 cells, following treatment for 1 hour. These results point to cytotoxic effects exerted by the butanolic extract from the roots of Pfaffia paniculata on human breast tumor cell lines

Considerando que diversas atividades biológicas dos ácidos clorogênicos já foram descritas na literatura, uma melhor compreensão da biossíntese e do acúmulo desses polifenois nas plantas está diretamente vinculada aos parâmetros de qualidade de fitoterápicos. Este trabalho teve o objetivo de monitorar a variação na concentração de metabólitos secundários do grupo dos ácidos clorogênicos no tecido foliar de diversas espécies diante de diferentes tratamentos pós-coleta. Este monitoramento foi realizado através da quantificação das substâncias por cromatografia líquida de ultra eficiência acoplada à espectrometria de massas em sequência (CLUE-EM/EM). Na primeira fase do estudo, foram coletadas folhas de cada espécie selecionada para o estudo (Caju - Anacardium occidentale L.; Graviola - Annona muricata L.; Pata-de-Vaca - Bauhinia variegata L.; Limão - Citrus limon (L.) Burm. f.; Café - Coffea arabica L.; Pitanga - Eugenia uniflora L.; Alecrim - Rosmarinus officinalis L., e Jambolão - Syzygium cumini (L.) Skeels). As folhas foram submetidas a um processo de extração e os extratos obtidos foram analisados por CLAE-EM/EM. Foram encontradas nove substâncias monossubstituídas nas posições 3, 4 e 5 pertencentes aos subgrupos dos ácidos p-cumaroilquínicos, cafeoilquínicos e feruloilquínicos dos ácidos clorogênicos, além de ácidos dicafeoilquínicos. A identificação dos sinais cromatográficos foi realizada por meio dos padrões de fragmentação de cada composto. Na segunda fase do estudo, foram coletadas quatro folhas dos indivíduos em estudo. As folhas foram congeladas em nitrogênio líquido (para interrupção do metabolismo) em tempos diferentes: imediatamente após a coleta (T0), 30 min após a coleta (T0,5), 1 h após a coleta (T1) e 2 h após a coleta (T2). Este procedimento foi realizado uma vez por mês durante seis meses. Em outro momento foram retiradas seis folhas de três indivíduos de espécies selecionadas (Alecrim, Pitanga, Pata-de-Vaca, Limão e Café) e cada folha foi congelada nos tempos T0 (imediatamente após a coleta), T6 (6 h após a coleta), T12 (12 h após a coleta), T24 (24 h após a coleta), T48 (48 h após a coleta) e Ts (após secagem das folhas - aproximadamente 30 dias após a coleta). O material coletado foi extraído e analisado por CLUE-EM/EM, no modo MRM. O método analítico quantitativo foi validado, considerando os parâmetros de linearidade, exatidão e precisão. As concentrações dos ácidos clorogênicos em estudo não apresentaram um padrão de variação que se relacione com os tempos de tratamento ou um aumento linear nas respostas dentro do intervalo de 2 h após o início do catabolismo do tecido. As amostras com tempos de tratamento mais longo, por sua vez, apresentaram aumento significativo de diversos compostos nas folhas que não tiveram seu metabolismo interrompido 30 dias após a coleta. Este acúmulo de ácidos clorogênicos corrobora a hipótese de que o aumento da concentração destes compostos pode estar relacionado ao catabolismo de polímeros fenólicos de maior massa. / Considering that diverse chlorogenic acids\' biological activities have been described, a better understanding of their biosynthesis and accumulation is closely related to phytotherapics\' quality parameters. This research aimed to monitor the variation on the concentration of secondary metabolites from the chlorogenic acids\' group on the leaf\'s tissue of diverse species, towards different post-harvest treatments. The monitoring was performed through the quantification of the compounds by UPLCMS/ MS. On the first phase of the project, leaves from each selected species (Cashew - Anacardium occidentale L.; Soursop - Annona muricata L.; Pata-de-Vaca - Bauhinia variegata L.; Lemon - Citrus limon (L.) Burm. f.; Coffee - Coffea arabica L.; Cherry - Eugenia uniflora L.; Rosemary - Rosmarinus officinalis L., and Jambolan - Syzygium cumini (L.) Skeels) were harvested. This material was submitted to an extraction process, and the obtained extracts were analyzed by HPLC-MS/MS. Nine substances were found, monossubstituted on positions 3-, 4- and 5-, belonging to the subgroups of p-coummaroylquinic, caffeoylquinic and feruloylquinic acids, in addition to dicaffeoylquinic acids. The chromatographic signals\' identification was preceded observing the fragmentation pattern of each compound. On the second phase of the study, four leaves were collected from each individual. The leaves were frozen with liquid nitrogen (to interrupt the metabolism) on different moments - immediately after the collection (T0), 30 min after de collection (T0,5), 1 h after the collection (T1) and 2 h after the collection (T2). This procedure happened once a month during six months. On August, 2016, a different collection was performed for a broader observation: six leaves were harvested from three individuals from selected species (Rosemary, Cherry, Pata-de-Vaca, Limon and Coffee), and each leave was frozen on T0 (immediately after the collection), T6 (6 h after the collection), T12 (12 h after the collection), T24 (24 h after the collection), T48 (48 h after the collection) and Ts (after leaf\'s drying - approximately 30 days after collection). The harvested material was extracted and analyzed by UPLC-MS/MS, on MRM mode. The analytical method was validated for linearity, accuracy and precision. Chlorogenic acids\' concentrations did not show a pattern of variation that related to the different treatments or a linear increase on response until 2 hours after the beginning of the tissue catabolism. The samples submitted to a broader treatment interval, however, showed a significant rise on the concentration of different compounds, on leaves that did not have their metabolism interrupted until 30 days after the collection. This accumulation of chlorogenic acids agrees with the hypothesis that the raise on the concentration of these substances can be related to the catabolism of polymeric phenols of higher mass.

Cordia ecalyculata Vell. (Boraginaceae), espécie medicinal vulgarmente conhecida como porangaba e chá-de-bugre, é utilizada popularmente como emagrecedor, diurético, para curar tosses catarrais e reumatismos além de ser usada no tratamento de úlceras externas em forma de banhos. No Brasil existe grande confusão com relação aos nomes populares das espécies medicinais, fato que motivou a realização do presente trabalho. A espécie estudada é confundida com outras espécies vegetais, particularmente com Casearia sylvestris Swartz (Flacourtiaceae), também conhecida pelos nomes de guaçatonga e erva-de-bugre. O estudo farmacobotânico de C. ecalyculata revelou estruturas papilosas na epiderme e células esclereificadas no parênquima cortical, que não haviam sido descritas anteriormente. Foram listadas as principais diferenças morfoanatômicas entre as folhas de C. sylvestris e C. ecalyculata. auxiliares na diagnose da droga vegetal comercializada. As amostras adquiridas no comércio como porangaba não apresentaram características de C. ecalyculata, mas sim, de C. sylvestris, como: duas a três camadas de parênquima paliçádico; cavidades secretoras; drusas e cristais prismáticos em abundância; estômatos paracíticos; células epidérmicas poligonais; ausência de litocistos e areia cristalina. Fotomicrografias ilustram o trabalho. A triagem fitoquímica do extrato hidroetanólico liofilizado (EHEL) de C. ecalyculata apresentou resultado positivo para taninos, flavonóides e saponinas. A análise cromatográfica permitiu verificar a presença de substância coincidente com a alantoína no extrato de C. ecalyculata, além de possibilitar o desenvolvimento do perfil cromatográfico, auxiliar na diferenciação de C. sylvestris. Os ensaios de quantificação de alantoína no EHEL foram realizados por meio de dois métodos espectrofotométricos, cujos valores encontrados foram 0,68% e 0,70% de alantoína. O EHEL não apresentou róxicidade no ensaio de toxicidade aguda, na dose de 5 g/kg de massa corpórea do animal, por via oral. O experimento de atividade antiúlcera do EHEL de C. ecalyculata não apresentou diferenças significativas entre os animais controle e tratados. / Cordia ecalyculata Vell. (Boraginaceae) is a medicinal species most known as porangaba and chá-de-bugre. It is commonly used as diuretic, weight controller, for phleem cough healing, rheumatism healing and also in baths for externai ulcers treatment. The present work aims at clarifying the popular names of medicinal species often misnamed in Brazil. For example, the species cited above has always been mistaken as Casearia sylvestris Swartz. (Flacourtiaceae), also known as guaçatonga and erva-de-bugre. The pharmacological-botanical study of C. ecalyculata revealed papillary structures on the epidermis and sclerenchymatous cells on the cortical parenchyma which had not yet been described in the literature. The main morphological-anatomical differences between the C. sylvestris and the C. ecalyculata leaves were listed. These differences have been of great help concerning the diagnosis of commercialized vegetal drug. The commercialized samples do not show the features of the C. ecalyculata, but the features of the C. sylvestris, such as: two or three palisade parenchyma layers; secretive cavities; plenty of druses and prismatic crystals; paracytic stomata; polygonal epidermical cells; absence of lythocysts and crystal-sand, as it can be seen in the illustrative photomicrographies. The phytochemical screening of the lyophilized hydroethanolical extract (EHEL) of the C. ecalyculata showed a positive outcome for tannins, flavonoids and saponins. The chromatographic analysis not only allowed the evaluation of the coincident substance with the allantoin in the C. ecalyculata extract, but also enable the development of its chromatographic aspect, what eventually helped to differenciate it from C. sylvestris. The quantification assays of allantoin in the EHEL extract of the C. ecalyculata were performed through two spectrophotometric methods and the values found were 0,68% and 0,70% of allantoin. The EHEL extract displayed no toxicity in the acute toxicity trial in the 5 g/kg oral dosage per animal body weight. The anti-ulcer assay of this extract showed no meaningful differences among the subject and the treated animais.

Au cours des dernières décennies, un certain nombre de brevets déposés sur des médicaments à base de plantes se sont vus dénoncés comme des cas de « biopiratage ». Ils sont en effet accusés de constituer une appropriation indue des connaissances de peuples autochtones sur les vertus thérapeutiques de plantes. Il est vrai qu'en droit positif, les produits phytopharmaceutiques peuvent répondre aux conditions légales de la brevetabilité, quand les savoirs médicinaux ne font l'objet d'aucun mécanisme de réservation. En réaction à ces pratiques prolifèrent donc des initiatives visant à définir les instruments juridiques qui permettraient de faire bénéficier les peuples autochtones des fruits de l'exploitation de leurs connaissances. Le droit international s'est ainsi récemment enrichi d'accords visant à la protection des savoirs traditionnels, parmi lesquels la Convention sur la Diversité Biologique de 1992, prolongée par le Protocole de Nagoya en octobre 2010. Les principes posés doivent cependant être traduits en des règles concrètes pour être mis en œuvre. C'est pourquoi, à différents niveaux, les acteurs proposent voire développent leurs propres instruments pour tenter d'apporter une réponse au problème du biopiratage, à la fois dans et en dehors du système de protection de la propriété intellectuelle. Ce sont les modalités d'introduction de ces différents mécanismes en droit positif qu'il s'agit ici d'étudier. Pour ce faire, l'analyse économique du droit est convoquée pour évaluer les tenants et les aboutissants de réformes qui, pour se trouver à la frontière entre les droits de la propriété intellectuelle, de l'environnement, du développement et du commerce international, soulèvent des questions juridiques imprégnées de considérations éthiques, politiques et économiques. / Over the past decades, several patents granted for plant-based medicine have been denounced as “biopiracy” based on the assertion that they misappropriated indigenous people's knowledge over plants' therapeutic properties. Indeed, in the current state of law, while the pharmaceuticals may be the subject of patent law, medicinal knowledge is not the subject of any proprietary regime. Therefore, many initiatives have been suggested that aim at defining the appropriate legal instruments to ensure that a sharing of the benefits is imposed on pharmaceutical companies when utilising medicinal knowledge. The principle that the traditional knowledge associated with medicinal plants requires legal protection has already found expression in a number of political outputs, including such major international agreements as the “Convention on Biological Diversity” signed in 1992. This has been further developed with the Nagoya Protocol adopted in October 2010. However, the principles in these conventions still need to be translated into a concrete legal framework to be fully enforceable. This is why it is equally important to study legal practice to see how various actors can explore ways of dealing with the biopiracy issue, both within and outside the existing intellectual property system. This study assesses the legal accuracy of these posited options at the intersection of intellectual property, environmental, development and international trade laws. It uses an economic analysis of law to evaluate the background and the outcomes of reforms that raise not only legal, but also economic, political, and ethical burning issues.

Thesis (MTech (Horticultural Sciences))--Cape Peninsula University of Technology, 2017. / The increasing demand for medicinal plants has led into serious over-harvesting of wild populations and presents an opportunity for potential profitable cultivation. Production of medicinal plants in controlled environments particularly hydroponic technology provides opportunities for high quality biomass accumulation and optimizes production of secondary metabolites. Water availability and supplies are becoming scarce, thus search for innovative irrigation practices is desirable and vital. The proper irrigation interval and growing media can play a major role in increasing the water use efficiency. Thus, Siphonochilus aethiopicus was cultivated by means of the hydroponic technique, under various substrate combinations and watering regimes.

Yip Pui Ying. / Thesis submitted in: November 2004. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 147-184). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iii / Acknowledgment --- p.v / Abbreviations --- p.vii / Table of contents --- p.viii / List of Figures --- p.xii / List of Tables --- p.xvii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1. --- The importance of characterization of Chinese medicinal materials and the development of Chinese medicine in Hong Kong --- p.1 / Chapter 1.2. --- Methods for characterization of Chinese medicinal materials --- p.5 / Chapter 1.3. --- Molecular characterization of Chinese medicinal materials --- p.8 / Chapter 1.3.1. --- DNA sequencing --- p.9 / Chapter 1.3.2. --- DNA fingerprinting --- p.14 / Chapter 1.3.3. --- Nucleic acid hybridization --- p.19 / Chapter 1.4. --- Objectives --- p.20 / Chapter Chapter 2 --- Characterization of Plant and Fungal Materials by rDNA ITS Sequence Analysis --- p.22 / Chapter 2.1. --- Introduction --- p.22 / Chapter 2.2. --- Materials and Methods --- p.22 / Chapter 2.2.1. --- Chinese medicinal materials used in this study --- p.22 / Chapter 2.2.1.1. --- Plants and fungi for interspecific ITS study --- p.22 / Chapter 2.2.1.2. --- Plant for intraspecific ITS study and locality study --- p.33 / Chapter 2.2.2. --- Extraction of total DNA --- p.35 / Chapter 2.2.3. --- PCR amplification of ITS1 and ITS2 regions of rRNA gene --- p.35 / Chapter 2.2.4. --- Purification of PCR products --- p.38 / Chapter 2.2.5. --- Cloning using pCR-Script´ёØ Amp SK(+) Cloning Kit --- p.38 / Chapter 2.2.5.1. --- Polishing --- p.38 / Chapter 2.2.5.2. --- Ligation of inserts into pCR-Script´ёØ Amp SK(+) cloning vector --- p.38 / Chapter 2.2.5.3. --- Transformation --- p.40 / Chapter 2.2.5.4. --- PCR screening of white colonies --- p.40 / Chapter 2.2.5.5. --- Purification of PCR screening products --- p.41 / Chapter 2.2.6. --- Sequencing of ITS regions --- p.41 / Chapter 2.2.6.1. --- Cycle sequencing reaction --- p.41 / Chapter 2.2.6.2. --- Purification of sequencing extension products --- p.41 / Chapter 2.2.6.3. --- Electrophoresis by genetic analyzer --- p.42 / Chapter 2.2.7. --- Sequence analysis and alignment --- p.42 / Chapter 2.3. --- Results --- p.42 / Chapter 2.3.1. --- Extraction of total DNA --- p.42 / Chapter 2.3.2. --- PCR amplification of ITS1 and ITS2 regions of rRNA gene --- p.44 / Chapter 2.3.2.1. --- Interspecific ITS study --- p.44 / Chapter 2.3.2.2. --- Intraspecific ITS study --- p.46 / Chapter 2.3.3. --- Sequence analysis and alignment --- p.47 / Chapter 2.3.3.1. --- Interspecific ITS study --- p.47 / Chapter 2.3.3.2. --- Intraspecific ITS study --- p.56 / Chapter 2.4. --- Discussions --- p.60 / Chapter 2.4.1. --- rDNA regions used for studying Chinese medicinal materials --- p.60 / Chapter 2.4.2. --- The results agreed with previously published works --- p.60 / Chapter 2.4.3. --- Explanation of interspecific results within the Ganoderma genus --- p.60 / Chapter 2.4.4. --- Implications from interspecific comparisons --- p.60 / Chapter 2.4.5. --- Implications from intraspecific comparisons --- p.61 / Chapter Chapter 3 --- .Characterization of Astragalus membranaceus by DNA Fingerprinting / Chapter 3.1 --- Introduction --- p.62 / Chapter 3.2 --- Materials and Methods --- p.62 / Chapter 3.2.1 --- Extraction of total DNA --- p.62 / Chapter 3.2.2 --- Generation and detection of DNA fingerprints by AP-PCR --- p.63 / Chapter 3.2.3 --- Analysis of DNA fingerprints --- p.63 / Chapter 3.3 --- Results --- p.63 / Chapter 3.3.1 --- Generation of DNA fingerprints by AP-PCR --- p.63 / Chapter 3.3.2 --- Fingerprint analysis --- p.69 / Chapter 3.4 --- Discussion --- p.85 / Chapter 3.4.1 --- RP-PCR has been used on Chinese medicinal materials --- p.85 / Chapter 3.4.2 --- AP-PCR used instead of RAPD --- p.85 / Chapter 3.4.3 --- Reproducibility and amount of bands --- p.86 / Chapter 3.4.4 --- Alternatives of electrophoresis process --- p.88 / Chapter 3.4.5 --- Explanation of results --- p.88 / Chapter 3.4.6 --- Distinguishing Neimengu and Shanxi samples --- p.89 / Chapter 3.4.7 --- Further studies --- p.90 / Chapter Chapter 4 --- Characterization of Plant and Fungal Materials by DNA-DNA Hybridization on Microarrays --- p.91 / Chapter 4.1 --- Introduction --- p.91 / Chapter 4.2 --- Materials and Methods --- p.92 / Chapter 4.2.1 --- Samples for microarray study --- p.92 / Chapter 4.2.2 --- Extraction of total DNA --- p.95 / Chapter 4.2.3 --- Amplification and sequencing of ITS 1 region of rRNA gene --- p.95 / Chapter 4.2.4 --- Preparation of labeled probe --- p.95 / Chapter 4.2.5 --- Amplification of ITS1 fragments --- p.97 / Chapter 4.2.6 --- Preparation of slides --- p.103 / Chapter 4.2.7 --- Hybridization and washing --- p.104 / Chapter 4.2.8 --- Scanning and data analysis --- p.105 / Chapter 4.3 --- Results --- p.105 / Chapter 4.3.1 --- DNA extraction --- p.105 / Chapter 4.3.2 --- Amplification and sequencing of ITS1 region of rRNA gene --- p.107 / Chapter 4.3.3 --- Preparation of labeled probe and amplification of ITS1 fragments… --- p.112 / Chapter 4.3.4 --- Preparation of slides --- p.112 / Chapter 4.3.5 --- Scanning and data analysis --- p.116 / Chapter 4.4 --- Discussion --- p.134 / Chapter 4.4.1 --- Implications --- p.134 / Chapter 4.4.2 --- Applying the findings --- p.134 / Chapter 4.4.3 --- Ways to maximize specificity --- p.137 / Chapter 4.4.4 --- Optimisation --- p.138 / Chapter 4.4.5 --- Microarray may be more advantageous over sequencing --- p.138 / Chapter Chapter Five --- General Discussion and Summary --- p.140 / Chapter 5.1. --- Objectives of this study --- p.140 / Chapter 5.2. --- rDNA ITS sequencing --- p.140 / Chapter 5.2.1. --- Description of the approach and summary of the results --- p.140 / Chapter 5.2.2. --- Implications from the results --- p.140 / Chapter 5.2.3. --- Advantages and limitations of DNA sequencing --- p.141 / Chapter 5.3. --- AP-PCR fingerprinting --- p.141 / Chapter 5.3.1. --- Description of the approach and summary of the results --- p.141 / Chapter 5.3.2. --- Advantages and limitations of DNA fingerprinting --- p.142 / Chapter 5.4. --- DNA-DNA hybridization on microarrays --- p.143 / Chapter 5.4.1. --- Description of the approach and summary of the results --- p.143 / Chapter 5.4.2. --- Implications from the results --- p.143 / Chapter 5.4.3. --- Advantages and limitations of DNA hybridization on microarrays. --- p.144 / Chapter 5.5. --- Overall summary --- p.144 / Chapter 5.6. --- Future studies --- p.146 / References --- p.147 / Appendix --- p.185

Objective To evaluate the cytotoxic and antibacterial effect of Bixa orellana L. (B. orellana) (achiote) methanol extract against Streptococcus mutans (ATCC 25175) (S. mutans) and Streptococcus sanguinis (ATCC 10556) (S. sanguinis). Methods Two methanol extracts of B. orellana were prepared in vitro, from the seeds and leaves. The antibacterial activity of extracts against S. mutans and S. sanguinis was evaluated using the cup-plate agar diffusion method. The minimum inhibitory concentration (MIC) was determined using the microdilution method and the cytotoxic activity was determinated by using the cell line MDCK. Results A stronger antibacterial effect was observed with the leaves methanolic extract with an inhibition zone of (19.97 ± 1.31) mm against S. mutans and (19.97 ± 1.26) mm against S. sanguinis. The methanolic extract of the seeds had an activity of (15.11 ± 1.03) mm and (16.15 ± 2.15) mm against S. mutans and S. sanguinis, respectively. The MIC of the leaf and the seed extracts against S. sanguinis was 62.5 and 125 μg/mL, respectively, and the MIC of the leaf extract against S. mutans was 62.5 μg/mL, and for the seed extract it was 31.25 μg/mL. The 50% cytotoxic concentration was 366.45 and 325.05 μg/mL for the leaves and seeds extracts, respectively. Conclusions The experimental findings demonstrated the antibacterial effect of the methanolic extract of B. orellana (achiote) on S. mutans and S. sanguinis. The extract of this plant is cytotoxic at high concentrations. / Peer review

O uso de plantas medicinais no tratamento de doenças tem aumentado de forma significativa nos últimos anos, bem como as pesquisas sobre a caracterização química destas plantas. Neste estudo, foram determinados os constituintes inorgânicos presentes nas folhas e nos extratos de três espécies da planta do gênero Casearia (C. sylvestris, C. decandra e C. obliqua) coletadas em regiões distintas da Mata Atlântica, SP. As composições elementares dos solos em que estas plantas foram cultivadas também foram determinadas. Tradicionalmente, estas plantas são utilizadas devido às suas propriedades antiinflamatórias, antiácidas, anti-sépticas e cicatrizantes. Além disso, as atividades antiulcerogênicas e antitumorais do gênero Casearia, e sua capacidade de neutralizar venenos de serpentes e abelhas, também tem sido comprovadas cientificamente. A metodologia analítica empregada foi a análise por ativação com nêutrons. Irradiações longas e curtas das amostras e dos padrões foram realizadas utilizando fluxos de nêutrons do reator de pesquisas IEA-R1 do IPEN/CNEN-SP. Nas folhas, o elemento K foi encontrado em níveis de porcentagens, os elementos Ca, Cl, Mg e Na na ordem de mg g-1, Br, Fe, Mn, Rb e Zn em μg g-1 e os elementos As, Co, Cr, Cs, La, Sb, Sc e Se em teores mais baixos, da ordem de ng g-1. Os resultados obtidos nas análises dos extratos indicaram que os mesmos elementos determinados nas folhas de Casearia foram também encontrados em seus respectivos extratos. A comparação das composições elementares das folhas de Casearia sylvestris coletadas em três regiões diferentes da Mata Atlântica mostrou que as concentrações dos elementos encontrados nas plantas dependem da composição dos solos em que elas foram cultivadas. Por outro lado, espécies diferentes do gênero Casearia cultivadas em uma mesma região apresentaram composições elementares similares. Estes resultados mostram a importância da realização de estudos que avaliem as atividades farmacológicas de Casearias cultivadas em diferentes tipos de solos. A qualidade analítica dos resultados foi avaliada pela análise dos materiais de referência certificados NIST 1573a Tomato Leaves, NIST 1515 Apple Leaves, INCT-TL-1 Tea Leaves, IAEA-Soil-7 e USGS W1. Os desvios padrão relativos e os erros relativos obtidos nestas análises indicaram boa precisão e exatidão dos resultados. / The use of medicinal plants in the treatment of diseases has increased significantly in the last years, as has research concerning chemical characterization of these plants. In this study, inorganic constituents were determined in leaves and in extracts from three medicinal plant species of the Casearia genus (C. sylvestris, C. decandra and C. obliqua) collected in distinct regions of the Atlantic Forest, SP. The elemental compositions of the soils in which these plants were grown were also determined. Traditionally, these plants are used due to their antiinflamatory, antiacid, antiseptic and cicatrizing properties. The antiulcer and the antitumor activities of the Casearia genus and its capacity to neutralize snake and bee venoms, have also been scientifically confirmed. The analytical methodology used was neutron activation analysis. Long and short irradiation periods of the samples and the standards were carried out at IPEN’s IEA-R1 nuclear research reactor. In the leaf K was found at the percentage levels, Ca, Cl, Mg and Na at mg g-1 levels and the elements Br, Fe, Mn, Rb and Zn at the μg g-1 levels. As, Co, Cr, Cs, La, Sb, Sc and Se at the ng g-1 levels. Results obtained in the extracts indicated that the same elements present in the leaves are also found in their extracts. The comparison between the inorganic composition of Casearia sylvestris leaves collected from three different regions of the Atlantic Forest showed that the elemental concentrations in the plants leaves varied depending on the place where they were grown. Different Casearia species cultivated in a same region presented similar elemental compositions. Based on these findings it can be concluded that the studies about the pharmacological effect of Casearia genus plants grown in different types of soil are of great importance. The quality of the obtained results was assured by the analyses of the certified reference materials NIST 1573a Tomato Leaves, NIST 1515 Apple Leaves, INCT-TL-1 Tea Leaves, IAEA-Soil-7 and USGS W1. The relative standard deviations and the relative errors obtained in these analyses indicated good precision and accuracy of the results.

Orientador: Hérida Regina Nunes Salgado / Banca: Hérida Regina Nunes Salgado / Banca: Eloir Paulo Schenkel / Banca: Taís Maria Bauab / Resumo: As espécies Vismia guianensis (Aubl.) (Clusiaceae) e Senna reticulata (Willd.) Irwin & Barneby (Caesalpiniaceae), conhecidas popularmente por lacre e mata-pasto, respectivamente, são encontradas na região amazônica, onde são utilizadas na medicina popular para o tratamento de diversas moléstias, principalmente infecções e dermatomicoses. Diante do exposto, o presente trabalho objetivou avaliar a atividade antimicrobiana dos extratos e frações das referidas espécies vegetais, através da bioautografia e da determinação da concentração inibitória mínima (CIM) pelo método de microdiluição. Foi avaliada também a atividade imunológica dos extratos hidroetanólicos das cascas do caule e das folhas de Senna reticulata e de Vismia guianensis através da produção e inibição de óxido nítrico por células peritoneais de camundongos. Todos os extratos apresentaram atividade inibitória frente às cepas padrão (ATCC) e clínicas dos fungos Trichophyton rubrum, Trichophyton mentagrophytes e Microsporum canis. O extrato hidroalcoólico das cascas de V. guianensis apresentou melhor atividade antimicrobiana, com CIM de 62,5 μg/mL, de 125 μg/mL e de 500 μg/mL frente às cepas padrão de T. mentagrophytes, T. rubrum e M. canis, respectivamente; CIM de 125 μg/mL para as cepas clínicas de T. rubrum e T. mentagrophytes e, de 15,63 μg/mL frente à cepa clínica de M. canis. O extrato hidroetanólico das cascas de V. guianensis também apresentou atividade antimicrobiana contra todas as bactérias testadas, sendo as cepas de S. aureus, S. epidermidis e B. subtilis foram as mais sensíveis. As cepas de C. albicans foram resistentes aos extratos de ambas espécies vegetais. Através dos ensaios de atividade imunológica in vitro observou-se que na concentração de 50 μg/mL os extratos não apresentaram citotoxicidade e que apresentaram potencial atividade antiinflamatória / Abstract: The Vismia guianensis (Aubl.) (Clusiaceae) and Senna reticulata (Willd.) Irwin & Barneby (Caesalpiniaceae), popularly known as seal and obtusifolia, respectively, are found in the Amazon region, where they are used in folk medicine for the treatment of various diseases, especially infections and dermatomycoses. In this light, this study aimed to evaluate the antimicrobial activity of extracts and fractions of these plant species by bioautography and determination of minimum inhibitory concentration (MIC) by microdilution method. We also evaluate the immunological activity of the hydroethanolic from the stem bark and leaves of Senna reticulata and Vismia guianensis and production through the inhibition of nitric oxide by peritoneal cells of mice. All the extracts showed activity against the standard strains (ATCC) and clinical fungi Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum canis. The hydroalcoholic extract of the bark of V. guianensis showed better antimicrobial activity, with MIC of 62.5 μg/mL, 125 μg/mL and 500 μg/mL against the standard strains of T. mentagrophytes, T. rubrum and M. canis, respectively, MIC of 125 μg/mL for clinical strains of T. rubrum and T. mentagrophytes and of 15.63 μg/mL against the clinical strain of M. canis. The hydroalcoholic extract of the bark of V. guianensis also showed antimicrobial activity against all bacteria tested, with strains of S. aureus, S. epidermidis and B. subtilis were the most sensitive. The strains of C. albicans were resistant to the extracts of both species. Through the in vitro testing of immune activity was observed that the concentration of 50 μg/mL extracts showed no cytotoxicity and showed potential anti-inflammatory activity / Mestre

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