Orientador: Adriana Zerlotti Mercadante / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-11T06:49:41Z (GMT). No. of bitstreams: 1 Ferreira_DanielaSouza_M.pdf: 861449 bytes, checksum: 0972445ef423fb371289eeef23009060 (MD5) Previous issue date: 2008 / Resumo: Dentre as várias opções de espécies frutíferas com boas perspectivas de comercialização, surge a amora-preta (Rubus spp.) como umas das mais promissoras. A amora-preta é uma pequena fruta que tem apresentado sensível crescimento nos últimos anos no Rio Grande do Sul, Sul de Minas Gerais e tem elevado potencial para ser cultivada no estado de São Paulo. No Rio Grande do Sul, a amora-preta tem tido grande aceitação pelos produtores, devido ao seu baixo custo de produção, facilidade de manejo, rusticidade e pouca utilização de defensivos agrícolas. Muitos fitoquímicos presentes em amora-preta exibem propriedades benéficas à saúde, como compostos fenólicos, com destaque para os pigmentos antociânicos. Estes pigmentos, que conferem a coloração atraente à fruta, possuem baixa estabilidade frente a algumas condições do meio como pH neutro e alcalino, alta temperatura e presença de luz. Assim, através deste estudo foram determinados espectrofotometricamente o teor de alguns compostos bioativos presentes em amora-preta cultivar Tupy, como antocianinas totais, monoméricas, poliméricas e copigmentadas, além de compostos fenólicos totais, flavonóides totais e carotenóides. A atividade antioxidante foi avaliada frente aos radicais ABTS e DPPH. Por cromatografia líquida de alta eficiência (HPLC), acoplada em série a detectores de arranjo de diodos (PDA) e de espectrômetro de massas (MS) foram identificadas as antocianinas e os carotenóides presentes no extrato de amora. Com o intuito de aumentar a estabilidade das antocianinas, o extrato antociânico da amora-preta foi encapsulado utilizando a técnica de gelificação térmica. O material de parede selecionado foi a curdlana por não perder a capacidade gelificante em pH abaixo de 2. Após o processo de encapsulação foram avaliadas algumas características das partículas como o tamanho médio, a eficiência de encapsulação e o perfil de liberação do recheio. As antocianinas identificadas em amora-preta foram cianidina 3-glucosídeo, cianidina 3-dioxalil-glucosídeo, cianidina 3-malonil-glucosídeo e cianidina 3- rutinosídeo. A antocianina majoritária foi a cianidina 3-glucosídeo perfazendo 92,9 % da área total. O teor de antocianinas totais foi de 90,5 ± 0,1 mg/100 g, sendo composto por 76,2 ± 0,3 % de monoméricas, 22,8 ± 0,4 % de poliméricas e 1,6 ± 0,1 % de copigmentadas. As antocianinas monoméricas foram encontradas como 104,1 ± 1,7 mg em cianidina 3-glucosídeo/100 g de fruta. Foi observado que a amora-preta possui baixo teor de carotenóides (86,5 ± 0,1 mg/100 g) e os carotenóides encontrados foram: all-trans-b-caroteno (39,6 %), all-trans-luteína (28,2 %), all-trans-b-criptoxantina (13,9 %), 9-cis-b-caroteno (3,8%), all-trans-a-caroteno (3,3 %), 13-cis-b-criptoxantina (3,1 %), all-transzeaxantina (2,7 %), 13-cis-b-caroteno (1,7 %), 5,6-epóxi-b-criptoxantina + fitoeno (0,8%), além de um carotenóide não identificado representando 2,9 %. Com a presença de elevados teores de compostos fenólicos totais (241,7 ± 0,8 mg/100 g) e de flavonóides totais (173,7 ± 0,7 mg/100 g), pode-se concluir que estes compostos presentes na amora-preta foram os principais responsáveis pela elevada capacidade antioxidante avaliada pela habilidade de capturar radicais livres ABTS (TEAC 2209,7 ± 68,4 mM/ g) e DPPH (EC50 33,8 ± 1,8 g amostra/g DPPH). As partículas formadas por gelificação térmica com 4,3, 5,1 e 5,6 % de curdlana apresentaram forma esférica e multinucleada. A distribuição de tamanho apresentou perfil semelhante para as diferentes concentrações de curdlana, embora as partículas obtidas com maior concentração de curdlana (5.6 %) apresentaram maior uniformidade no tamanho, maior umidade e maior eficiência de encapsulação. O perfil de liberação de todas as partículas contendo antocianinas apresentou cinética exponencial de primeira ordem, com total liberação nos primeiros 20 minutos em tampão pH 1,0 / Abstract: Among the fruit species with good perspectives for commercialization, blackberry fruit (Rubus spp.) is one of the best options. Blackberry is a small fruit, which has been increasingly cultivated in Rio Grande do Sul State, South of Minas Gerais State, also presenting high potential for cultivation in São Paulo State. In Rio Grande do Sul, blackberry has been widely accepted by the farmers, due to its low production cost, easy handling, rusticity and use of low amounts of agricultural defensives. Several phytochemicals found in blackberry, such as phenolic compounds and anthocyanins, show beneficial health properties to humans. The anthocyanins, responsible for the attractive color of blackberries, possess low stability in some medium conditions, such as neutral and alkaline pH, high temperature and presence of light. Thus, using spectrophotometric methods, the levels of some bioactive compounds were determined in blackberries cv. Tupy, such as total anthocyanins, monomeric, polymeric and copigmented anthocyanins, as well as phenolic compounds, total flavonoids and total carotenoids. The antioxidant activity was determined using the free radicals ABTS and DPPH. Both anthocyanins and carotenoids from blackberry extracts were separated and identified by high performance liquid chromatography (HPLC) connected in series to diode array (PDA) and mass spectrometer (MS) detectors In order to increase the anthocyanin stability, the anthocyanic blackberry extract was encapsulated by thermal gelification. Curdlan was the wall material selected, since the gel formed was stable in pH values lower than 2. After the encapsulation process, some particle characteristics, such as medium size, encapsulation efficiency and release profile, were evaluated. The anthocyanins identified in blackberry were cyanidin 3-glucoside, cyanidin 3-dioxalyl-glucoside, cyanidin 3-malonil-glucoside and cyanidin 3-rutinoside. The major anthocyanin was cyanidin 3-glucoside, representing 92.9 % of the total area. The levels of total anthocyanins were 90.5 ± 0.1 mg/100 g, composed by 76.2 ± 0.3 % of monomeric, 22.8 ± 0.4 % of polymeric and 1.6 ± 0.1 % of copigmented ones. The monomeric anthocyanins were quantified as 104.1 ± 1.7 mg of cyanidin 3-glucoside/100 g of fruit. The levels of carotenoids found in blackberry were low (86.5 ± 0.1 mg/100 g), represented by all-trans-b-carotene (39.6 %), all-trans-lutein (28.2 %), all-trans-b-cryptoxanthin (13.9 %), 9-cis-b-carotene (3.8%), all-trans-a-carotene (3.3 %), 13-cis-b-cryptoxanthin (3.1 %), all-trans-zeaxanthin (2.7 %), 13-cis-b-carotene (1.7 %), 5,6-epoxy-b-cryptoxanthin + phytoene (0.8%), and a not identified carotenoid representing 2.9 %. The elevated levels of phenolic compounds (241.7 ± 0.8 mg/100 g) and flavonoids (173.7 ± 0.7 mg/100 g) are most probably responsible for the high antioxidant activity against the free radicals ABTS (TEAC 2209.7 ± 68.4 mM/100g) and DPPH (EC50 33.8 ± 1.8 g sample/g DPPH). The particles formed by thermal gelification with 4.3, 5.1 and 5.6 % of curdlan presented spherical and multinucleated forms. The profiles of the size distribution were similar for all curdlan concentrations; although the particles with the highest curdlan concentration (5.6 %) showed highest size distribution uniformity, highest moisture and highest encapsulation efficiency. The release profile of anthocyanins from all particles followed first-order kinetics, with total release in 20 minutes in buffer pH 1.0 under agitation at room temperature / Mestrado / Mestre em Ciência de Alimentos

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-07-11T13:40:59Z No. of bitstreams: 1 2012 - Luzimary de Jesus Ferreira Godinho Rocha.pdf: 1344154 bytes, checksum: 0659f7d53a1501d32b67bcaa4079a4da (MD5) / Made available in DSpace on 2017-07-11T13:40:59Z (GMT). No. of bitstreams: 1 2012 - Luzimary de Jesus Ferreira Godinho Rocha.pdf: 1344154 bytes, checksum: 0659f7d53a1501d32b67bcaa4079a4da (MD5) Previous issue date: 2012-07-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The pitaya or Cereus undatus pitaya or, synonymy: Hylocereus guatemalensis, H.undatus is exotic fruit consumption and slightly increasing in our country. The functional assignments given to this fruit, common sense encourages the study of their physical, chemical and microbiological. It should be noted that the fruits are primary sources of several vitamins and other bioactive compounds, for example, phenolic fibers, and sugars. The intake of these compounds increases the immunity of individuals, leading to improved levels of health, physical and mental performance. The reference values for these nutrients pitaya, also, are unknown to the general public, as this is a fruit intake of a wealthy class, for its price is too high for our Brazilian standards. In food matrices are very complex, given its inherent characteristics. Thus, there are several techniques used for quantitative analysis of bioactive compounds, among them, have a High Performance Liquid Chromatography (HPLC) and UV-visible spectrophotometry. The objective of this study is to quantify the presence of vitamin C, anthocyanins and sugars in pitayas white and red pulp by HPLC, as well as to determine, although the content of antioxidant activity by the method of capturing the radical 2,2 '- azinobis ( 3-etilbenzenotiazolina-6-sulfonic acid - ABTS), soluble solids (? Brix) is determined in a digital refractometer, as well as acidity and pH. The soluble solids found can confirm that the samples of red pitaya have higher sugar content than the white pulp. The pH and total acidity (g citric acid/100g fruit), should also be monitored and analyzed, possibly under conditions of controlled cultivation of this food because it is an exotic fruit and consumption in our country recently. As for the results of anthocyanins, because it is a dye that rapidly degrades, its presence became insignificant, being found only in the halo of red pulp pitaya, requiring further analysis and monitoring standards in a more specific. There was a low antioxidant activity in the samples, as well as its content of vitamin C, these values should be found, first, the storage time, which decreases the levels of these analytes. / A pitaya ou Cereus undatus, sinon?mia: Hylocereus guatemalensis, H.undatus ? uma fruta ex?tica e de consumo ligeiramente crescente no nosso pa?s. As atribui??es funcionais dadas a essa fruta, pelo senso comum, incita ao estudo das suas caracter?sticas f?sicas, qu?micas e microbiol?gicas. Deve-se ressaltar que as frutas s?o fontes prim?rias de v?rias vitaminas e outros compostos bioativos, como por exemplo, os compostos fen?licos, fibras e a??cares. A ingest?o desses compostos aumenta a imunidade dos indiv?duos, induzindo a melhoria dos n?veis de sa?de, rendimento f?sico e mental. Os valores de refer?ncia desses nutrientes para a pitaya, ainda, s?o desconhecidos do grande p?blico, por ser esta uma fruta de consumo de uma classe abastada, por seu pre?o ser demasiadamente alto para os nossos padr?es brasileiros. As matrizes em alimentos s?o muito complexas, dadas as suas caracter?sticas intr?nsecas. Diante disso, v?rias s?o as t?cnicas utilizadas para determina??es anal?ticas de compostos bioativos, dentre elas, t?m-se a Cromatografia L?quida de Alta Efici?ncia (CLAE) e espectrofotometria U.V vis?vel. O objetivo deste trabalho ? quantificar a presen?a de vitamina C, antocianinas e a??cares nas pitayas de polpas branca e vermelha por CLAE, bem como, determinar, ainda, o teor de atividade antioxidante pelo m?todo de captura do radical 2,2?- azinobis (3-etilbenzenotiazolina-6-?cido sulf?nico ? ABTS), teores de s?lidos sol?veis (?Brix) determinado em um refrat?metro digital, al?m da acidez e pH. O teor de s?lidos sol?veis encontrados pode confirmar que as amostras de pitaya vermelha t?m maiores teores de a??cares que a de polpa branca. Quanto ao valor de pH e acidez total titul?vel (g de ?cido c?trico/100g de fruta), precisam, ainda, ser monitorados e analisados, possivelmente, sob condi??es de cultivo controlado desse alimento por se tratar de uma fruta ex?tica e de recente consumo no nosso pa?s. Quanto aos resultados das antocianinas, por se tratar de um corante que degrada rapidamente, a sua presen?a mostrou-se irris?ria, sendo encontrada somente no halo da pitaya de polpa vermelha, necessitando de mais an?lises e padroniza??es em condi??es mais espec?ficas de monitoramento. Houve baixa atividade antioxidante nas amostras analisadas, bem como o seu teor de vitamina C, esses valores encontrados devem-se, primeiramente, ao tempo de armazenamento, que diminui os teores desses analitos.

O presente trabalho teve como foco principal, a preparação one pot de compostos orgânicos alquílicos e vinílicos contendo o grupamento butiltelurenila em sua estrutura, e o estudo do comportamento desses em variados tipos de reações. Uma delas foi a reação de troca Te/Li. Assim, teluretos foram utilizados como equivalentes sintéticos de vários reagentes organometálicos. Os teluretos foram submetidos à reação de troca Te/Li, e os respectivos compostos organolítio gerados, transformados em reagentes organometálicos de zinco, cobre e cério através de reações de transmetalação. Esses reagentes gerados in situ foram submetidos à reação com variados eletrófilos. Os reagentes organometálicos de zinco e cobre gerados foram submetidos à reação de acilação frente a cloreto de ácidos, resultando em 1,4-hidroxicetonas. Um exemplo dessas hidroxicetonas foi obtido em sua forma enantiopura utilizando um telureto quiral, e esse utilizado na síntese formal e enantiosseletiva da (-)-Pirenoforina. Diferentes estequiometrias de cupratos e outros reagentes organometálicos oriundos de um telureto também foram preparados, e esses testados em reações de substituição de tosilatos e abertura de epóxidos. Os substratos gerados dessas reações foram empregados na síntese das moléculas bioativas Endo-Brevicomina e Frontalina. Apresentaremos também, o resultado obtido na reação direta entre uma espécie di-litiada quiral na presença de benzonitrila. A hidroxicetona assim produzida é um análogo do Ipomeanol, um composto com atividade contra câncer de pulmão. Será apresentado também, o comportamento de entidades di-aniônicas contendo grupamentos silila em reações catalisadas por CeCl3, em que a captura desses com eletrófilos como aldeídos, forneceram 1,4-enedióis em bons rendimentos químicos e alta diastereosseletividade. Será apresentado também o estudo do comportamento de teluretos com hibridização sp2 em reações pericíclicas. A reação pericíclica estudada compreende compostos contendo telúrio (II) e (IV) em reações de Diels-Alder. / One of the main purposes of this work was to develop a new direct methodology to prepare functionalized organic tellurides, to be submitted to a series of different reactions, as for example the Te/Li exchange reaction, aimed to afford synthetic equivalents of several organometallic reagents. Zinc, copper and cerium organometallic compounds were generated via a transmetalation reaction of the lithium species, and were reacted with several electrophiles. As a result, 1,4-hydroxyketones could be successfully prepared. According to this methodology, an enantioenriched 1,4-hydroxyketone could be obtained, using a chiral telluride as starting material. This chiral nonracemic intermediate was employed in an enantioselective formal synthesis of (-)-Pyrenophorin. Cuprates and other organometallic reagents, prepared from a specific telluride and using the above described methodology, were the choosen nucleophiles for performing some aliphatic substitution reactions and, in particular, for the ring opening of epoxides. The resulting products were employed as building blocks for the synthesis of the bioactive cyclic compounds Endo-Brevicomin and Frontalin. The direct reaction of the chiral dilithiated specie with benzonitrile afforded a 1,4-hydroxyketone as an analogue of Ipomeanol, a bioactive compound for cancer therapy. Bis-anionic sililated species, prepared from a telluride, were submitted to reaction with a series of aldehydes. Such reactions, in which CeCl3 was employed as catalyst, showed to be highly diastereoselectives, affording (E)-1,4-enediols in good yields. The pericyclic reaction of insaturated tellurides was also investigated. It is worth mentioning that the same Diels-Alder reaction could be performed either with tellurium II or tellurium IV species.

Orientador: Marcos José Salvador / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-25T08:29:51Z (GMT). No. of bitstreams: 1 Andreazza_NathaliaLuiza_D.pdf: 7725533 bytes, checksum: 49ffd2e5015cd199c9f7b084991b2d1e (MD5) Previous issue date: 2014 / Resumo: Neste trabalho realizou-se estudo físico, químico e biológico para a prospecção de metabólitos secundários e fitoderivados em Guatteria blepharophylla (Annonaceae) e Indigofera truxillensis (Fabaceae) para aplicação em Terapia Fotodinâmica (PDT). A caracterização fitoquímica dos extratos metanólicos das espécies estudadas resultou no isolamento de cinco alcaloides isoquinolínicos de G. blepharophylla e um alcaloide bis-indólico de I. truxillensis. A investigação fotofísica dos extratos, frações e das substâncias isomoschatolina e índigo, revelou perfis de absorção na região entre 600 a 800 nm. Nas análises fotoquímicas, os extratos e fração alcaloídica de G. blepharophylla apresentaram resultados sugestivos quanto a produção de oxigênio singleto, assim como a substância isomoschatolina. Já em ensaio microbiológico in vitro com o emprego de substâncias supressoras de espécies reativas de oxigênio, observou-se a prevalência do mecanismo fotoquímico do tipo II para a isomoschatolina e do tipo I para o índigo. Nos ensaios biológicos de PDT antimicrobiana e antitumoral com os extratos, frações e as substâncias isoladas isomoschatolina e índigo observaram-se, para todas as estas amostras-teste, resultados efetivos na inativação de algumas das cepas indicadoras com redução do crescimento microbiano (UFC/ml) superior a 90%, assim como diminuição da sobrevida celular para as linhagens humanas de fibroblasto (3T3) e melanoma (UACC-62), sendo observado efeito em menores concentrações somente para UACC-62 quando associado à irradiação laser (660nm). Em ensaio biológico na presença das substâncias CaCl2 e MgCl2, a ação biocida dos alcaloides isomoschatolina e índigo, na fotoinativação da cepa Escherichia coli ATCC 10799 foi intensificada se comparado aos grupos controles. Portanto o conjunto de resultados observados sugerem que os extratos brutos e frações alcaloídicas das duas plantas estudadas e os alcaloides isomoschatolina e índigo apresentaram fotoatividade frente a bactérias, leveduras e frente à cultura de células de melanoma em concentração subinibitória. Na análise da interação dos alcaloides fotossensibilizantes berberina e isomoschatolina com LDL em modelos celulares, a caracterização dos complexos alcaloide/LDL permitiu identificar uma primeira classe de fixação, denominada classe P, a partir de alterações na fluorescência da apoproteína B-100. Para berberina e isomoschatolina verificou-se que cerca de 250 e 135 moléculas se complexam a porção proteica da partícula de LDL e proximidades, apresentando constante de afinidade iguais a 1,05.108 M-1 e 5,06.107 M-1, respectivamente. Uma segunda classe de fixação, chamada de classe L, determinada somente para a berberina, foi caracterizada a partir de alterações na fluorescência deste alcaloide e correspondeu a fixação de cerca de 80 moléculas de berberina na porção lipídica da partícula de LDL, com uma constante de afinidade de 7,10.107 M-1. Estes valores revelam que o processo predominante de interação da berberina com o LDL é a fixação de classe P. Nos ensaios em nível celular, realizados com a berberina, verificou-se que a sua complexação com o LDL não prejudica o seu reconhecimento pelos receptores apoproteína B/E da superfície membranar das células U87MG e que a sua internalização pode ocorrer associada o LDL pela via de endocitose. No entanto, não foi verificado acumulo de berberina nos lisossomos, mas apenas nas mitocôndrias, o que sugere que após a internalização, o alcaloide rapidamente sofre um processo de redistribuição intracelular. Para células cultivas em Ultroserum G, observou-se aumento da produção de produtos da peroxidação lípidica nas células tratadas com berberina complexada em relação àquelas tratadas com o alcaloide livre, nas doses de irradiação de 50 e 100 J/cm2. Todas estas evidências revelam a possibilidade de complexação de ambos os alcaloides com o LDL como molécula carreadora para aplicação em PDT antitumoral, sendo que para berberina a associação não só aumenta sua incorporação celular, assim como sua efetividade como agente fotoativo / Abstract: In this work it was carried out chemical and biological studies for secondary metabolites and phyto-derivatives prospecting from Guatteria blepharophylla (Annonaceae) and Indigofera truxillensis (Leguminosae) for antimicrobial and antitumor photodynamic therapy (PDT) application. The phytochemical characterization of the methanol extracts from both species resulted in the isolation of five isoquinoline alkaloids from G. blepharophylla, isomoschatoline, O-methylmoschatoline, liriodenine, subsessiline and lysicamine, and one bis-indole alkaloid, indigo, from I. truxillensis. Photophysical characterization of crude extracts, fractions and the substances isomoschatoline and indigo showed absorption profiles in the region between 600 to 800 nm and fluorescence emission when excited at their absorption wavelenght. In photochemical analysis, at 1,3-DPBF probe assay, crude extracts and alkaloidal fraction from G. blepharophylla showed positive results regarding singlet oxygen production, as well as isomoschatoline. At in vitro microbiological assay with reactive oxygen species suppressors, it was observed the prevalence of type II photochemical mechanism for isomoschatoline and type I for indigo, as also suggested by 1,3- DPBF assay results. In the antimicrobial and antitumor PDT biological assays, with crude extracts, fractions, isomoschatoline and indigo, effective results for microbial strains inactivation were found with microbial growth reduction (CFU/mL) superior to 90%, as well as decreased cell survival of human fibroblast (3T3) and melanoma (UACC-62) cell lines, in which this effect for the latter, was observed at lower concentrations when combined with laser irradiation. It was also found that both isomoschatoline and indigo exhibit enhanced photodynamic antimicrobial activity against Gram-negative bacteria (Escherichia coli ATCC 10799 strain) in comparison to control groups, when CaCl2 and MgCl2 additives were employed. Therefore, in light of all assays results it is suggested that crude extracts and alkaloid fractions from both plant species and the alkaloids isomoschatoline and indigo showed photoactivity against bacteria and yeasts strains and against melanoma human cell line at sub-inhibitory concentration when associated with laser irradiation at 660 nm. At interaction studies between photosensitizing alkaloids berberine and isomoschatoline with LDL and cellular models, changes in LDL¿s Apoprotein B-100 fluorescence allowed the identification of a first class of substances fixation in this particle called, P fixation class. For berberine and isomoschatoline it was found that about 250 and 135 molecules, respectively complexes to LDL¿s protein portion and its vicinity, leading to affinities constants equals to 1,05.108 M-1 and 5,06.107 M-1, respectively. A second fixation class, the L fixation class, accessed only for berberine, was obtained by changes in the alkaloid fluorescence and correspond to 80 molecules attached to LDL¿s lipid portion, leading to affinity constant equal to 7,10.107 M-1. These results reveal that the predominant process of berberine interaction with LDL is by P fixation class. At cellular level assays, performed only with berberine, it was found that the alkaloid complexation with LDL did not affect LDL recognition by apoprotein B/E membrane surface receptors of U87MG cells and that berberine internalization may occur associated with the particle via LDL endocytosis. However, there was no accumulation of berberine found in lysosomes. Mitochondria accumulation was observed suggesting that, after internalization, this alkaloid undergoes rapid intracellular redistribution process. At cells cultured with Ultroserum G and treated with complexed berberine it was also observed an increase in lipid peroxidation in comparison with cells incubated with free BBR, under irradiation doses of 50 to 100 J/cm2. All these evidences indicate the possibility of both alkaloids complexation with LDL as a carrier molecule for antitumor PDT application and, for berberine, this association not only increases its cellular uptake, as well as its effectiveness as photoactive agent / Doutorado / Biologia Vegetal / Doutora em Biologia Vegetal

Orientador: Gabriela Alves Macedo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-26T09:06:48Z (GMT). No. of bitstreams: 1 Barbosa_PauladePaulaMenezes_M.pdf: 2172137 bytes, checksum: 707d48554bb74b873d31d80aa96163e6 (MD5) Previous issue date: 2014 / Resumo: Compostos bioativos são moléculas orgânicas derivadas de vegetais, animais ou micro-organismos que estão relacionados à alguma atividade biológica. Os micro-organismos do solo representam uma importante fonte de compostos biologicamente ativos. O ponto crítico na descoberta de novas moléculas bioativas desta fonte é o isolamento de grupos de micro-organismos pouco explorados e conhecidos que são ao mesmo tempo bons produtores de metabólitos secundários. Os fungos filamentosos são conhecidos como produtores de uma grande variedade de metabólitos secundários. Diante disso, o objetivo deste trabalho foi isolar e selecionar linhagens de fungos filamentosos silvestres, de duas regiões do Estado de São Paulo (Biomas de Mata Atlântica e Cerrado), com vistas para produção de compostos antimicrobianos ativos contra os micro-organismos alvo escolhidos nesta pesquisa: Staphylococcus aureus, Bacillus cereus, Escherichia coli, Salmonella choleraesuis e Candida albicans. Estes micro-organismos causam grandes problemas na área de alimentos e saúde. A partir das amostras de solo coletadas na região de Mata Atlântica (Ilhabela) e na região de transição dos biomas Mata Atlântica e Cerrado (Barão Geraldo, Campinas), 118 culturas de fungos filamentosos foram isoladas. Dentre elas 22 apresentaram elevado potencial antimicrobiano contra pelo menos um dos micro-organismos alvo e foram estudadas quanto ao seu efeito antimicrobiano. Ao final, foram obtidas culturas de fungos filamentosos com elevado potencial para produção de metabólitos com atividade antimicrobiana, os quais poderão ser identificados e explorados futuramente / Abstract: Bioactive compounds are organic molecules from vegetables, animals or microorganisms that are related to some biological activity. Soil microorganisms represent an important source of biologically active compounds. The isolation of unexplored and unknown microorganisms that are both good producers of secondary metabolites, provides the potential for novel bioactive molecules. Filamentous fungi are known as good producers of a great variety of secondary metabolites. Therefore, this work goal was to isolate and select wild filamentous fungi strains of two regions of São Paulo State (Biomes of Cerrado and Atlantic Forest), capable to produce antimicrobial compounds against the microorganisms: Staphylococcus aureus, Bacillus cereus, Escherichia coli, Salmonella choleraesuis and Candida albicans. These microorganisms cause important problems in food and health areas. 118 filamentous fungi strains were isolated from soil samples collected at Atlantic Forest region (Ilhabela) and transition region between Atlantic Forest and Cerrado biomes (Barão Geraldo, Campinas). Among them, 22 strains showed high antimicrobial activity against at least one of the pathogenic microorganisms target and their antimicrobial effect were studied. In conclusion, potentially filamentous fungi strains producers of antimicrobial activity metabolites were obtained, which can be identified and future explored / Mestrado / Ciência de Alimentos / Mestra em Ciência de Alimentos

M.Sc. / The continued use and popularity of plant based traditional medicine necessitates scientific validation of the therapeutic potential of medicinal plants through phytochemical and pharmacological screening as well as the isolation and identification of bioactive compounds. Since the pharmacological effectiveness of medicinal plants is affected by several intrinsic and extrinsic factors, studies on the variations on chemical composition and biological activity are important as well. These provide a scientific rationale for using plants from different regions for the same medicinal purposes and allow traditional healers and consumers to make informed decisions with regard to the collection and use. Rapanea melanophloeos is a popular medicinal plant from the Myrsinaceae family widely distributed in southern Africa. It‟s bark, fruits and rarely the leaves are used traditionally for ailments ranging from stomach disorders, respiratory problems to disorders of the nervous system. Available chemical information reports on the accumulation of benzoquinones as major compounds, as well as some triterpenoid saponins and tannins. In view of the plant‟s wide distribution and medicinal use of different plant parts, this study comparatively evaluated the chemical composition of various crude extracts of the leaves, fruits and bark of plants collected from six localities. This was coupled with antibacterial tests to evaluate the therapeutic potential of different solvent extracts of the leaves, fruits and bark as well as the isolation of bioactive compounds from the fruits. Plants were collected from six different localities between Swaziland and South Africa and sequentially extracted with petroleum ether, ethyl acetate, methanol and water as a series of increasingly polar solvents. Thin layer chromatography (TLC), Gas-Chromatography/Mass spectrometry (GC-MS) and High Performance Liquid Chromatography (HPLC) were used to obtain a semi-quantitative chemical composition profile of different extracts. The TLC fingerprints of petroleum ether, ethyl acetate and methanol extracts showed the accumulation of non-polar terpenes, benzoquinones, saponins, tannins and flavonoids in the three plant parts. Slight variations in the chemical composition of the leaves, bark, and fruits were noted. Some components occurred in specific plant parts and others occurred in all three plants. The chemical profile of the leaves and bark were quite comparable in most instances with the fruits showing a generally different profile. This confirms previous literature reports on the comparability of leaves and the bark accounting for interchangeable use in traditional medicine.

Strawberries can be considered a functional food because their consumption has been associated with several health benefits. They are important sources of bioactive compounds, such as vitamins and polyphenolic compounds, with recognized antioxidant capacity (AOC). However, strawberry overall quality and bioactive content are greatly affected by environmental conditions during pre- and post-harvest and, little is known about the stability of its bioactive compounds, specifically ascorbic acid (AA) and polyphenolics compounds. Furthermore, additional research that addresses the impact of polyphenolic compounds on in vitro and in vivo models is needed to understand the mechanisms behind their potential health benefits. Therefore, the objectives of the work presented in this thesis were to: 1) evaluate the impact of different disease control treatments on strawberry bioactive compounds and AOC; 2) understand the relationship between bioactive compounds and AOC in strawberries and fruit juices; 3) investigate the origin of AOC in strawberries by identifying their major polyphenolic compounds and, 4) explore the effects of polyphenol-rich fruits and fruit juices on the proliferation of cancer cells and lifespan of Caenorhabditis elegans. Conscientious consumers are aware of the health benefits of substantial fruit and vegetable consumption but are also concerned about the amount of pesticide residues that can be found in conventionally grown produce, with pesticide-free produce (i.e., organic) becoming more popular. However, the market price for organic strawberries can be more than twice that of conventionally grown fruit which discourages the average American from purchasing this fruit on a regular basis. Therefore, in the first study presented in this thesis, we hypothesized that reducing pesticide usage would provide the consumer with a “sustainable strawberry” that would have better or similar quality at a lower cost than organic fruit while it would also reduce environmental impact and risk to pesticide applicators. Results from this study showed that strawberries from a reduced fungicide treatment, had better or similar bioactive content and AOC than fruit from the conventional disease control treatment. After cold storage, strawberries from the reduced or conventional disease control treatments showed comparable amounts of bioactive compounds and AOC. These results indicate that growing strawberries with a reduced number of fungicide applications can be an alternative to the conventional disease control or organic practices as it may reduce residual fungicides in the fruit, decrease production costs while still retaining important bioactive compounds. In order to understand the relationship between bioactive compounds and AOC in strawberries and fruit juices, 56 different types of commercial beverages were chosen for the second study presented in this thesis. Overall, results showed that the higher the total phenolic contents (TPC) in the beverage the higher their AOC. Amongst all beverages studied, aronia, blackcurrant, and pomegranate juices contained the highest amount of TPC and AOC. Furthermore, after opening the bottles, these juices were maintained for 14 days at 4 °C, to test the stability of their TPC which was in general relatively stable throughout storage. Further investigation on individual polyphenolic compounds and their possible contribution to the overall AOC of fruits and fruit juices, led to a third study. Overall, results showed that the AOC of major individual polyphenolic compounds found in strawberries (i.e., pelargonidin, cyanidin, ellagic acid, quercetin, kaempferol, catechin, epicatechin, caffeic acid, p-coumaric acid, ferulic acid) was significantly higher than that of mixtures of the same compounds. In addition, the AOC of strawberries correlated with its major bioactive compounds (i.e., polyphenolic compounds and ascorbic acid) in a form of a synthetic bioactive strawberry model (“Powerberry”) composed of major strawberry polyphenolic compounds, vitamin C, fructose and glucose in the same ratios found in a real strawberry. These results suggest that even though strawberries contain many different polyphenolic compounds and vitamins, their AOC might only depend on few compounds that are found in significant quantities in the fruit. Finally, using cell and worm models we were able to demonstrate that conventional and organic strawberry, raspberry and blueberry fruits, and aronia, blackcurrant and pomegranate juices successfully inhibited the proliferation of HeLa cervical cancer cell lines. In addition, when introduced in low doses (0.75 mg ml-1 or lower) to the C. elegans diet, aronia, blackcurrant and pomegranate juices promoted longevity. Overall, results suggest that using whole fruit or fruit juices might constitute an alternative of treating cancer cells in vivo and that polyphenolic compounds contained in fruits and fruit juices displayed significant bioactivity in a worm model.

Stroke is a devastating and debilitating condition resulting from a blockage or hemorrhage in the vasculature of the brain. Despite extensive research, the etiology and pathophysiology of the disease at the level of the cell membrane are poorly understood, and effective treatment has been elusive. Though much research has shown marked increases in lipid metabolism following stroke, the impact of these changes have often been overlooked given the technical challenges associated with identifying regionally specific changes in degenerating tissue. The advent of lipidomics – a systems biology approach to the large-scale profiling of individual lipid species in tissues – has renewed interest in understanding the role of membrane lipids and their metabolites in the cell and in ischemic injury. In this thesis, I have used an unbiased LC-ESI-MS-based lipidomic approach to profile the small molecular weight glycerophosphocholine second messenger lipidome in anterior and posterior regions of cortex and striatum in the forebrain of wild-type and platelet activating factor receptor (PAFR) null-mutant mice before and after middle cerebral artery occlusion (MCAO). From these profiles, I have outlined the potential use of lipid second messenger distribution as topographic landmarks to identify functional subdomains within neural tissue. Further, I have demonstrated that ischemia does not simply disrupt lipid second messenger metabolism globally but produces regionally specific changes in discrete species and that these changes are altered by the loss of lipid regulatory effectors (i.e., PAFR null mutation). Based on the lipid species identified in this profile of healthy and ischemic tissue, I proposed that tight regulation of PC(O-22:6/2:0) homeostasis by PAFR-expressing microglia is ii required for proper dopaminergic signaling in prefrontal cortex. Finally, I have outlined a model whereby increased PAF synthesis following ischemia contributes the inflammatory response by promoting blood-brain barrier permeability, microglial activation and immune cell infiltration in a PAFR-dependent manner.

Orientador: Miriam Dupas Hubinger / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-27T10:07:35Z (GMT). No. of bitstreams: 1 Rabelo_RenataSantos_M.pdf: 54735801 bytes, checksum: 79deeea44988fbcfd818411f5b804573 (MD5) Previous issue date: 2015 / Resumo: A alcachofra (*Cynara scolymus L.*) é uma herbácea perene pertencente à família *Asteraceae. *A espécie, rica em compostos fenólicos, é originária de regiões subtropicais, mas também é cultivada no Brasil, onde maior parte da produção se concentra no interior paulista e é voltada à indústria de conservas*,* que geralmente descarta como resíduo, brácteas externas e internas da planta. Em vista disso, o presente trabalho teve como objetivo, desenvolver extratos ricos em compostos bioativos, principalmente fenólicos, a partir dos resíduos de alcachofra, por meio do estudo do processo de extração assistida por ultrassom e da avaliação do desempenho do processo de concentração por membranas. Inicialmente, foram estudadas as variáveis da extração hidroalcoólica assistida por ultrassom como potência do ultrassom, composição do solvente e razão sólido/solvente. Após esta etapa, dois dos extratos, nas melhores condições de extração, foram concentrados por nanofiltração, usando três membranas de poliamida (NF270, DK e DL), em um sistema de filtração tangencial operado a 20 bar. Os processos foram avaliados quanto ao rendimento de compostos fenólicos (Folin-Ciocauteau), ácido clorogênico (CLAE-DAD) e capacidade antioxidante (DPPH e FRAP). Os rendimentos obtidos com a integração das etapas de extração e concentração, a partir de um fator de redução de volume de 2,5 foram de 559,59 ± 32,09 mg de ácido gálico equivalente/g de extrato seco para fenólicos totais, e 22,95 ± 0,35 mg de ácido clorogênico/g de extrato seco, para ácido clorogênico. Os resultados para capacidade antioxidante foram de 198,88 ± 15,15 mg Trolox equivalente/g de extrato seco, para o método de DPPH, e de 385,70 ± 0,35 mg Trolox equivalente/g de extrato seco para o método FRAP. O declínio do fluxo de permeado no processo de nanofiltração foi causado pela formação de torta, e a membrana que apresentou menor resistência ao fluxo, maior estabilidade ao longo do processo de filtração e maior retenção dos compostos de interesse, foi a DK, sendo esta a mais indicada para concentração dos extratos avaliados nesse trabalho / Abstract: The artichoke (Cynara scolymus L.) is a perennial herb belonging to the family Asteraceae. The species, rich in phenolic compounds, is originally from subtropical regions, but is also cultivated in Brazil, where most of its production is concentrated in the interior of São Paulo, and is geared to the canning industry, where external and internal bracts of plant are usually discarded as waste. The present study aimed to develop extracts rich in bioactive compounds, mainly phenolics, from artichoke waste by the study of the extraction process assisted by ultrasound and the assessment of the performance of the membrane concentration process. Initially, the variables of water-alcohol extraction assisted by ultrasound (ultrasound power, solvent composition and solid/solvent ratio) were studied, after this, two extracts in the best extraction conditions were concentrated by nanofiltration, using three polyamide membranes (NF270, DK, DL), in a tangential filtration system operated at 20 bar. Process yields were evaluated for phenolic compounds (Folin-Ciocauteau), chlorogenic acid (HPLC-DAD) and antioxidant capacity (DPPH and FRAP). The yield obtained with the integration of extraction and concentration processes, from a volume reduction factor of 2.5 were of 559.59 ± 32.09 mg of gallic acid equivalent/g dry extract for total phenolics, and 22.95 ± 0,35 mg of chlorogenic acid/g of dry extract, for chlorogenic acid. The results for antioxidant capacity were of 198.88 ± 15.15 mg Trolox equivalent/g of dry extract to the DPPH method, and 385.70 ± 0.35 mg Trolox equivalent/g of dry extract for the FRAP method. The decline of the permeate flux in nanofiltration process is due to cake formation. The membrane that showed less resistance to flow, greater stability over the filtration process and more retention of the compounds of interest was DK, which is the most suitable for concentration of the extract evaluated in this work / Mestrado / Engenharia de Alimentos / Mestra em Engenharia de Alimentos

Os frutos da pitangueira pertencem à família Myrtaceae e podem ser encontrados em três variedades da mesma espécie, reconhecidos de acordo com a cor dos frutos como amarelo, vermelho e roxo. O objetivo deste estudo é caracterizar as três variedades de frutos de pitanga, com relação aos principais carotenoides e compostos fenólicos, e seu comportamento durante o desenvolvimento. Os frutos foram obtidos da região de Campinas (IAC) e fornecidos pela EMBRAPA - (RS). Os extratos metanólico dos frutos foram caracterizados quanto ao conteúdo antocianinas monoméricas, onde somente a variedade roxa, das diferentes plantas e região, apresentou aumento de concentração de antocianinas monoméricas com o desenvolvimento (0,59 a 1,94 mg cy-3-glu eq/g b.s. e 1,43 a 8,47 cy-3-glu eq/g b.s. para o estádio intermediário e maduro, respectivamente. Além disso, a variedade roxa no estádio maduro apresentou maior concentração de fenólicos totais (6,52 a 18,42 mg de AG/g b.s.) quando comparados às outras. O ácido cítrico foi identificado, por CLAE/DAD, como o ácido orgânico majoritário, seguido pelos ácidos succínico e málico, em todas as variedades, com maior concentração no estádio verde. De acordo com a concentração de ácido ascórbico, a pitanga não é uma boa fonte de vitamina C, apresentando valor máximo de 4,0 mg/g b.s no estádio maduro. Os açúcares majoritários encontrados foram a glicose e a frutose. A antocianina majoritária, identificada por LC-ESI-MS/MS, da variedade roxa, no estádio maduro foi a cianidina-3-glicosídeo, com uma variação de 8,05 a 640,53 mg/100g b.s, já o flavonoide majoritário na variedade amarela e vermelha foram miricetina e quercetina em ambas variedades. O carotenoide majoritário identificado por CLAE/DAD, em todas variedades no estádio maduro foi o licopeno, onde a variedade vermelha se destaca pela maior concentração. Dessa forma, o perfil de flavonoides não é alterado com o desenvolvimento, mas é observado uma redução no seu conteúdo nas variedades vermelha e amarela; já a variedade roxa apresenta alteração no perfil de flavonoides, com a expressão de antocianinas no estádio maduro. Por outro lado, o perfil de carotenoides em todas as variedades não sofreu alteração. / The fruits of the pitangueira belong to the family Myrtaceae and can be found in three varieties of the same species, recognized according to the color of the fruits as yellow, red and purple. The aim of this study was characterize the fruits of three pitanga varieties, according to the carotenoids and phenolic compounds composition, and their behavior during ripening. The fruits were obtained from the Campinas region and also provided by EMBRAPA - (RS). Monomeric anthocyaninswere detected only on purple variety, presenting 0.59 to 1.94 mg cy-3-Ο-glu eq/g d.w. and 1.43 to 8.47 cy-3-Ο-glu eq/g d.w.) for the intermediate and mature stages, respectively. In addition, the purple variety at the mature stage had the highest total phenolics concentration (6.52 to 18.42 mg of AG/g d.b.) as compared to other varieties. The citric acid was identified as the major organic acid, followed by succinic and malic acids, in all varieties, with a higher concentration in the green stage. According to the concentration of ascorbic acid, pitanga could not be considered a good source of vitamin C, presenting the highest amount of the 4.0 mg/g d.w. at the mature stage. The major soluble sugars found were glucose and fructose. The major anthocyanin of the purple variety, identified by LC-ESI-MS/MS in the mature stage, was cyanidin-3-Ο-glucoside, ranging from 8.05 to 640.53 mg /100g d.w., whereas the major flavonoids in the yellow and red varieties were myricetin and quercetin. The flavonoid profile was not altered with ripe developing, but a decrease in its content was observed in the red and yellow varieties; on the other hand, the purple variety presented a change in the flavonoid profile, with the expression of anthocyanins in the mature stage. The carotenoids profile in all varieties was the same. The major carotenoid identified by HPLC/DAD, at the mature stage, was lycopene, where the red variety presented the highest concentration.