Thesis (Ph. D.)--University of Canterbury, 2010. / Typescript (photocopy). Includes bibliographical references (p. 231-255). Also available via the World Wide Web.

Entre as frutas, a uva é uma das maiores fontes de compostos fenólicos, sendo que a indústria de derivados de uva gera uma grande quantidade de resíduos que, na maioria das vezes, são desprezados. Estes resíduos concentram quantidade significativa de compostos bioativos, quedespertam o interesse devido a suas propriedades antioxidantes e seus efeitos benéficos para a saúde humana. No intuito de aproveitamento deste subproduto, o presente estudo objetivou elaborar uma farinha utilizando resíduo de uva e analisar suas propriedades bioativas. Foram utilizadas duas técnicas de secagem do resíduo de uva (estufa e liofilizador) para a obtenção de dois tipos de farinhas, uma obtida pelo resíduo proveniente do sistema convencional de cultivo e outra proveniente do sistema orgânico de cultivo. Além disso, parte das uvas havia sido tratada com radiação UV-C. A farinha foi utilizada para elaboração de biscoitos tipo cookies, para avaliar o potencial como ingrediente alimentício e agregação de compostos bioativos no produto. Teores de antocianinas totais, compostos fenólicos totais e atividade antioxidante, expressa em EC50 e em TEAC foram determinados para o resíduo, farinha e biscoitos. Também foram determinados o pH, o teor de cinzas e cor para o resíduo, a farinha e o biscoito Em geral pode-se observar que o tratamento com UV-C apresentou efeito negativo ou nulo nos resultados. A liofilização demonstrou manter mais as propriedades bioativas. O sistema de cultivo convencional demonstrou uma maior manutenção das propriedades bioativas em parte das análises. / Among the fruits, the grape is one of the largest sources of phenolic compounds. the industry of grape spinoffs produces a large amount of waste that most often are underrated. These residues concentrate significant amount of bioactive compounds that arouse interest due to its antioxidant properties and its beneficial effects on human health. In order to take advantage of this by-product, the present study aimed to elaborate flour using residual grape and analyze their bioactive properties. Was used two techniques of drying the residue grape (dryer and freeze dryer) to obtain two kinds of flour, one obtained from the residue from the conventional cultivation and another derived from the organic cultivation. Furthermore, the grapes had been treated with UV-C radiation. The flour was used to prepare cookies, to evaluate its potential as a food ingredient and aggregation of bioactive compounds in the product. The contents of total anthocyanins, phenolic compounds and antioxidant activity, expressed as EC50 and TEAC were determined in the residue, fluor and cookies. Overall treatment with UV-C showed negative or null results. Lyophilization demonstrated longer maintain bioactive properties, as phenolic compounds and anthocyanins. Regarding the type of cultivation, the conventional system in some analyzes showed better maintenance bioactive properties.

Orientador: Lin Chau Ming / Banca : Regina Marta Evangelista / Banca: Pablo Forlan Vargas / Resumo: A batata-doce é uma das principais hortaliças cultivadas no mundo, as suas raízes tuberosas têm múltiplos usos na alimentção humana e animal e na produção industrial de farinha, amido e etanol. Os teores de compostos biativos, caracteristicas fisico-quimicas de quailidade pós-colheita e produtividade, podem diferir entre genótipos e idade da planta. O Vale do Ribeira é uma região de comunidades tradicionais que mantêm diferentes acessos de batata-doce. Este estudo foi realizado de fevereiro a outubro de 2015, o objetivo principal foi realizar a caracterização físico-química e de produtividade de três acessos de batata-doce pertencentes ao Banco de Germplasma da Universidade Estadual Paulista.Registro. As análises do experimento foram feitas nos acessos VR 13-01 (casca roxa e polpa roxa fortemente pigmntada), VR13-11 (casca rosa e polpa branca) e VR13-50 (casca creme e polpa branca). O delineamento experimental foi em blocos casualizados com seis repetições por tratamento e cada parcela tinha 5 plantas.O espaçamento de plantio entre plantas foi de 0,25 m e 0,80 m entre linhas. As avaliações das analises físico-químicas, compostos bioativos e produtividade foram feitas aos 120,150 e 180 dias após o plantio. Foram determinados a produçao total de matéria fresca da parte aérea, produçao total de matária seca da raiz, produtividade total de raizes, produtividade comercial de raízes, produção de raizes não comercias, teor de sólidos solúveis, teor de amido, comteúdo de composto... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract:The sweet potato is one the most grown vegetable crops in the world, its storage roots have multiple uses as human and animal foods and in industrial production of flour, starch and ethanol. The starch and sugar content may differ genotypes and harvest season. The Vale do Ribeira region has traditional communities keep different accessions of sweet potato. This study was conducted from February to October 2015, and main objetive was the physico-chemical characterization, bioactive compounds and productivity of three accessions of sweet potato belonging to the Germplasn Bank of Universidad Estadual Paulista. Accession were VR 13-01 (purple skin and strongly purple pulp).VR13-11( lightly purple skin and white pulp) and VR13-50 ( cream skin and white pulp).The experimental design was randomized blocks with six replications per treatment and each plot had 5 plants. The planting distance between plants was 0.25 m and 0.80 m between rows. The biochemical, physico chemical characterization and production yields was made 120,150 and 180 days after planting. The total productions yield, soluble solids content, and starch content were determined and significant differences among accessions obtained by ANOVA and means separated by a Scott-Knott test. Comparing all three accessions. VR13-01 showed highhest soluble solid content (16,6° Brix), anthocyanin content (111,6 mg 100-1 and accessions VR13-50 showed the highest starch content (25,3 %) and the highest production yields ... / Mestre

Orientador: Sarita Leonel / Banca: Maria Cecília de Arruda Palharini / Banca: Regina Marta Evangelista / Banca: Simone Rodrigues da Silva / Banca: Terezinha de Fatima Fumis / Resumo: O experimento foi realizado na Fazenda Experimental São Manuel, da Faculdade de Ciências Agronômicas da UNESP, Campus de Botucatu (SP), nos anos de 2014 e 2015. Objetivou-se avaliar o efeito da aplicação pré-colheita de cloreto de cálcio (CaCl2) na qualidade de frutos da amoreira preta 'Tupy'. Foram avaliadas plantas, cujas mudas foram plantadas em 23/06/2009. Conduziram-se seis hastes mais vigorosas, numa área experimental não irrigada, no sistema de espaldeira em T, com dois arames e com 1,2 metros de altura. O espaçamento foi de 0,6 m entre plantas e 4,0 m entrelinhas e a densidade de plantio de 4.166 plantas ha-1. No experimento o delineamento experimental adotado foi de blocos casualizados em fatorial 4 x 3, sendo o primeiro fator correspondente às concentrações de cloreto de cálcio (0; 1,5; 3 e 4,5 %) e o segundo aos números de aplicações (1, 2 e 3 aplicações). A fase fenológica adotada para a primeira aplicação do cloreto de cálcio foi dos frutos verdes em início de inchamento e ainda aderidos aos restos florais. Quando realizada mais de uma aplicação, havia um intervalo de cinco dias entre uma e outra. Foram realizadas avaliações físicas, físico-químicas, bioquímicas e enzimáticas nos frutos. Os níveis de Ca+2 foram aferidos nas folhas antes e após as aplicações com CaCl2 e nos frutos logo que realizada a colheita. A aplicação de CaCl2 em pré-colheita, na concentração de 4,5 %, quando os frutos estavam na fase fenológica de inchamento dos frutos com restos florais, pr... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The objective of this experiment was to evaluate the effect of pre-harvest application of calcium chloride (CaCl2) on 'Tupy' blackberry fruits, which was carried out at the São Manuel Experimental Farm, at the Faculty of Agronomic Sciences of UNESP, Botucatu Campus (SP), in the years 2014 and 2015. As the fruits of the black mulberry 'Tupy' plants were evaluated, whose seedlings were planted on 06/23/2009. Six more vigorous stems were conducted in a non-irrigated experimental area in the T-tier system with two wires and 1,2 meters in height. The spacing was 0,6 m between plants and 4,0 m between rows and the planting density of 4,166 plants ha-1. In the experiment, the experimental design was a randomized block in factorial 4 x 3, the first factor corresponding to the concentrations of calcium chloride (0, 1,5, 3 and 4,5 %) and the second to the numbers of applications (1, 2 and 3 applications). The phonological phase adopted for the first application of calcium chloride was of the green fruits at the beginning of swelling and still adhered to the flower remains. When more than one application was performed, there was an interval of five days between one application and another. Physical, physicochemical, biochemical and enzymatic evaluations were performed on the fruits. The levels of Ca+2 were measured in the leaves before and after the applications with CaCl2 and in the fruits as soon as the harvest was carried out. The application of pre-harvest CaCl2, at a concentration of 4,5 %, when fruits were in the phonological stage of fruit swelling with floral remains, promotes lower weight loss, higher firmness and ascorbic acid content in blackberries 'Tupy' ... / Doutor

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

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.