In vivo growth-inhibition of Sarcoma 180 by piplartine and piperine, two alkaloid amides from Piper

Piplartine {5,6-dihydro-1-[1-oxo-3-(3,4,5-trimethoxyphenyl)-2-propenyl]-2(1H)pyridinone} and piperine {1-5-(1,3)-benzodioxol-5-yl)-1-oxo-2,4-pentadienyl]piperidine} are alkaloid amides isolated from Piper. Both have been reported to show cytotoxic activity towards several tumor cell lines. In the present study, the in vivo antitumor activity of these compounds was evaluated in 60 female Swiss mice (N = 10 per group) transplanted with Sarcoma 180. Histopathological and morphological analyses of the tumor and the organs, including liver, spleen, and kidney, were performed in order to evaluate the toxicological aspects of the treatment with these amides. Administration of piplartine or piperine (50 or 100 mg kg-1 day-1 intraperitoneally for 7 days starting 1 day after inoculation) inhibited solid tumor development in mice transplanted with Sarcoma 180 cells. The inhibition rates were 28.7 and 52.3% for piplartine and 55.1 and 56.8% for piperine, after 7 days of treatment, at the lower and higher doses, respectively. The antitumor activity of piplartine was related to inhibition of the tumor proliferation rate, as observed by reduction of Ki67 staining, a nuclear antigen associated with G1, S, G2, and M cell cycle phases, in tumors from treated animals. However, piperine did not inhibit cell proliferation as observed in Ki67 immunohistochemical analysis. Histopathological analysis of liver and kidney showed that both organs were reversibly affected by piplartine and piperine treatment, but in a different way. Piperine was more toxic to the liver, leading to ballooning degeneration of hepatocytes, accompanied by microvesicular steatosis in some areas, than piplartine which, in turn, was more toxic to the kidney, leading to discrete hydropic changes of the proximal tubular and glomerular epithelium and tubular hemorrhage in treated animals.

Efeito hipoglicêmico de rações à base de berinjela (Solanum melongena,L.) em ratos

Os objetivos do presente trabalho foram verificar se dietas ricas em berinjela apresentam efeito hipoglicêmico, se contribuem para o retardamento de absorção da glicose pós-prandial e se a fração fibra solúvel está correlacionada com estes efeitos. Os experimentos foram realizados com ratos Wistar machos adultos, normais e diabéticos, que receberam rações à base de caseína, berinjela com casca e sem casca, e de casca de berinjela por um período de 42 dias. A glicose sangüínea foi determinada no início do experimento e aos 13, 21, 34 e 42 dias. Os testes orais de tolerância à glicose (TOTG) foram realizados no final do experimento. Os resultados mostraram que as rações à base de farinha de berinjela com casca e de casca de berinjela apresentaram redução nos níveis de glicose. Este efeito não foi significativo para a ração à base de farinha de berinjela sem casca. Os animais do grupo diabético que receberam ração à base de berinjela com casca apresentaram menor área sob a curva de glicose, do que os dos grupos controle (ração à base de caseína e de berinjela com casca) e do que o do grupo diabético que recebeu a ração de caseína. Este efeito não foi observado nos animais que receberam ração à base de berinjela sem casca e casca de berinjela. Estes resultados indicam a presença de um composto responsável pelo efeito hipoglicêmico na casca de berinjela e que a pectina solúvel da ração não foi suficiente para promover tal efeito, mas que a administração contínua de pectina solúvel contribuiu para a melhora da TOTG.

Determinação de glicoalcalóides em batatas in natura (Solanum Tuberosum L.) comercializadas na cidade de Campinas, Estado de São Paulo

A batata é uma das principais hortaliças cultivadas no Brasil, constituindo-se em um dos mais importantes alimentos na dieta humana, em decorrência de sua disponibilidade e características nutricionais. Entretanto, membros da família Solanaceae produzem, durante crescimento e após colheita, compostos potencialmente tóxicos denominados glicoalcalóides, dos quais alfa-solanina e alfa-chaconina predominam. A concentração máxima desses compostos em batata in natura considerada segura para consumo humano é estimada em 200mg·kg-1, expressa como glicoalcalóides totais (GAT). No presente estudo, foram extraídos e quantificados os GAT em amostras de tubérculos de batata in natura comercializados na cidade de Campinas, SP. A técnica utilizada foi a cromatografia líquida de alta eficiência (CLAE) com coluna C18 e detetor de arranjo de diodos. O valor médio de recuperação foi 102,6% para alfa-solanina e 100,2% para alfa-chaconina. Os limites de quantificação foram 2,5µg·mL-1 para alfa-solanina e 1,4µg·mL-1 para alfa-chaconina. As concentrações de GAT em amostras individuais de tubérculos inteiros das diferentes variedades e tipos estudados (Bintje, Monaliza, Asterix e Bolinha) variaram de 22,4 a 246,9mg·kg-1. Das amostras analisadas, 82% apresentaram níveis de GAT inferiores a 100mg·kg-1, o que indica que as variedades de batatas estudadas podem ser consideradas seguras para consumo humano.

Elaboração de biscoitos tipo salgado, com alto teor de fibra alimentar, utilizando farinha de berinjela (Solanum melongena, L.)

Devido ao fato da berinjela (Solanum melongena, L.) apresentar um alto teor de fibra alimentar e tendo em vista que biscoitos possuem grande potencial para servir como produto nutritivo, o presente estudo teve como objetivo utilizar a farinha de berinjela na obtenção de biscoitos tipo salgado para apresentarem alto teor de fibra e serem sensorialmente aceitáveis. Utilizaram-se níveis de 10, 15 e 20% de farinha de berinjela. Nos biscoitos prontos avaliaram-se as características químicas e físicas. Os teores de proteína, cinzas, extrato etéreo e fibra alimentar total, dos biscoitos elaborados, aumentaram com o incremento das proporções utilizadas. Por outro lado, a expansão e o peso unitário dos biscoitos diminuíram com o aumento da concentração de farinha de berinjela e sua coloração se tornou mais escura. Os biscoitos elaborados com os três níveis de farinha de berinjela podem ser classificados como produtos de alto teor de fibra alimentar e prontos para consumo.

Desenvolvimento e aceitabilidade de geléia dietética de cubiu (Solanum sessiliflorum Dunal)

Visando o aproveitamento de frutos amazônicos e atender à demanda crescente por novos produtos no mercado, foi formulada e avaliada a aceitabilidade e vida-de-prateleira da geléia de cubiu contendo xilitol em substituição à sacarose. Os frutos procedentes da Estação Experimental do Ariaú do Instituto Nacional de Pesquisas da Amazônia foram branqueados, despolpados, triturados, peneirados e acondicionados em embalagens plásticas à temperatura de -20 ± 1 °C até o momento da utilização. Concomitantemente, uma alíquota da polpa foi analisada quanto à umidade, proteínas, lipídios, fibras solúveis e insolúveis, cinza, pH, acidez total, açúcares totais e redutores, e compostos fenólicos. Para a formulação da geléia com uso de xilitol e convencional foi utilizada a proporção de 1:1 (polpa:xilitol e polpa:sacarose), respectivamente. A mistura foi concentrada até a obtenção de 65 °Brix , com ajuste de pH e adição de pectina. As geléias foram avaliadas quanto aos teores de proteínas, lipídios, cinza, pH, acidez, açúcares e compostos fenólicos e a vida-de-prateleira por meio de análises físico-química, microbiológica e sensorial, mensalmente, por um período de 180 dias. Os resultados demonstraram que não houve diferença estatística entre a aceitabilidade das geléias à base de xilitol e convencional, e que ambas apresentaram estabilidade físico-química e microbiológica durante o seu armazenamento. O xilitol pode ser uma alternativa viável em substituição à sacarose para o preparo de geléia de cubiu.

Efeito da substituição parcial da farinha de trigo por farinha de casca de batata (Solanum Tuberosum Lineu)

O aproveitamento dos subprodutos da agroindústria de alimentos diminui os custos da produção, aumenta o aproveitamento total do alimento e reduz o impacto que esses subprodutos podem causar ao serem descartados no ambiente. Dessa forma, alguns subprodutos da batata são aproveitados e transformados em ingredientes alimentícios, como é o caso da casca. Neste estudo utilizou-se a casca de batata para produção de farinha visando a sua utilização em produtos de panificação, sendo utilizadas as seguintes porcentagens na composição da farinha mista: 0-50-50%; 3-47-50%; 6-44-50%; 9-41-50% e 12-38-50% de farinha de casca de batata, farinha de trigo branca e farinha de trigo integral, respectivamente. Foram realizadas análises físico-químicas na farinha de casca de batata, análises físicas na farinha de casca, nas farinhas de trigo branca e integral e análises reológicas nas farinhas mistas. A farinha de casca de batata apresentou bons teores de fibras e minerais, e dentro deste último, destacam-se os teores de fósforo, cálcio e magnésio. Quanto à granulometria e à diferença de cor, as farinhas de casca de batata e a farinha de trigo integral apresentaram valores mais próximos uma da outra. À medida que se aumentou a substituição da farinha de trigo branca pela farinha de casca de batata, as análises reológicas indicaram uma queda na qualidade tecnológica das massas.

Physicochemical changes in cubiu fruits (Solanum sessiliflorum Dunal) at different ripening stages

Cubiu shrubs (Solanum sessiliflorum Dunal) have drawn the attention of researchers for their biological versatility (preferential heliophilous or facultative ombrophilous shrubs), their capacity to grow in upland or lowland areas, and the good technological quality of their fruits for the food industry. The aim of this study was to verify physicochemical changes in cubiu fruits during maturation. The fruits were harvested from the experimental station of olericulture of the Instituto Nacional de Pesquisas da Amazônia (INPA), Brazil. The analyses were performed in whole cubiu fruits (peel, pulp, and placenta) at four traditional ripening stages (green, turning, ripe, and fully ripe) for the determination of weight, moisture, total solids, total carotenoids, proteins, lipids, and ash. Cubiu fruits showed large weight variation, with amodal distribution. The ripe stage was critical to maintain moisture, and from that stage on, water loss became evident. The lipids increased steadily over the four ripening stages, maintaining, however, insignificant calorie content. Total carotenoids, proteins, and ash reached the maximum level at the fully ripe stage. With the exception of weight and moisture, all physicochemical changes exhibited the same general behavior, i.e. they increased as the fruits ripened at the four investigated stages.

Phenolic content and antioxidant capacity in organically and conventionally grown eggplant (Solanum melongena) fruits following thermal processing

AbstractThermal processing and production practices used in vegetables can cause changes in their phytochemical contents. Eggplant is characterized by its high antioxidant content. The objective of this work was to determine levels of anthocyanins, polyphenols, and flavonoids and antioxidant capacity in organically and conventionally grown eggplant prepared fresh or subjected to one of three thermal preparation methods: boiling, baking or steaming. The soluble and hydrolyzable polyphenols and flavonoids content were quantified by Folin-Ciocalteu and Aluminum chloride methods, respectively. Anthocyanins were quantified according to the pH differential method. Antioxidant capacity was determined by DPPH and ORAC methods. The results showed differences between organic and conventional eggplant for some variables although cultivation method did not have a consistent effect. Hydrolysable polyphenol content was greater, and soluble and hydrolysable antioxidant capacities were higher in organically grown eggplant, while anthocyanin content was greater in conventionally grown eggplant. Fresh eggplant produced under conventional cultivation had a much greater content of anthocyanins compared to that of other cultivation method-thermal treatment combination. In general, steamed eggplant contained higher total polyphenol and flavonoid levels as well as greater antioxidant capacity. Steamed eggplant from both conventional and organic systems also had high amounts of anthocyanins compared to other thermal treatments.

Influence of heat treatment on the sensory and physical characteristics and carbohydrate fractions of french-fried potatoes (Solanum tuberosum L.)

AbstractThe aim of this study was to analyze the impact that heat treatment with salts and freezing processes on the sensory, instrumental, and physico-chemical characteristics of fried potatoes of the Monalisa cultivar. The potatoes were blanched in distilled water (P); sodium chloride solution (B1); calcium chloride solution (B2), and a solution with both of these salts (B3). They were then pre-cooked and frozen for 24 hours and for 30 days. After frying, sensory characteristics were analyzed (color, texture, flavor, oiliness), along with overall preference and instrumental determinations of texture, color, and oil content. Further tests were conducted on the sample with the best results in the sensory analysis (B1), along with sample P as a control, to determine granule microstructure, carbohydrate fractions, glycemic index, and glycemic load. Blanching B3, despite reducing oil absorption and providing less oiliness, obtained lesser overall preference. Freezing for 30 days increased the lightness, except for when sodium chloride was used, which intensified the color yellow. The use of sodium chloride did not interfere with the type of starch granules, nor with the formation of resistant starch; however, longer freezing time reduced the glycemic index and concentrated the dietary fiber content. All samples exhibited low glycemic index and moderate glycemic loads.

Postharvest UV-B treatments increased fructose content of tomato (Solanum lycopersicon L. cv. Tayfun F1) harvested at different ripening stages

Abstract In the postharvest stage, taste and flavor are the key components of the marketability of tomato. Therefore, greater emphasis is now being placed on improving traits such as sugar content. In this study postharvest ultraviolet-B (UV-B) treatments on sugar, total soluble solids, and color of tomatoes harvested at different stages were investigated. Tomatoes harvested at turning, pink, and red stages were treated with two different doses of UV-B irradiation: UVB4 and UVB8. Color L* and hue angle values of tomatoes treated with UV-B were found to be high, which means the red color of tomatoes was improved. UVB4 treatments increased the color a* and saturation index values of tomatoes at pink and red harvest stage, although it did not affect at the turning stage. Additionally, UV-B irradiation treatments had no effect on sucrose content of the tomatoes. Fructose, glucose, and TSS content of tomatoes treated with UVB8 at red harvest stage were found to be high. Hence, the results obtained from this study are of great importance in terms of providing an increase in the amount of sugar without the need for breeding, and also consumer satisfaction.

Morfologia de frutos e sementes de espécies arbóreas do gênero Solanum L.

O objetivo do trabalho foi caracterizar a morfologia do fruto e da semente de três espécies florestais nativas do Brasil, Solanum granuloso-leprosum Dunal, S. lycocarpum A.St.-Hil. e S. pseudoquina A.St.-Hil., recomendadas para plantações destinadas à recuperação de áreas degradadas. Os frutos são indeiscentes, carnosos, do tipo baga, globosos, polispérmicos e constituídos por dois ou mais lóculos. As sementes são estenospérmicas, campilótropas, elipsóides, comprimidas, apresentando seção longitudinal largo-ovalada ou achatado-ovalada e seção transversal elíptica. Hilo mediano-marginal localizado em uma depressão e micrópila arredondada. Sementes albuminosas, com endosperma abundante, periférico, carnoso-firme, semitransparente e de coloração esbranquiçada. Embrião axial, linear, contínuo e curvado. Em Solanum lycocarpum e S. pseudoquina o embrião é circinado e em S. granuloso-leprosum é espiralado.

Bases para a padronização do teste de germinação em três espécies de Solanum L.

Foi objetivo deste trabalho obter informações básicas sobre a germinação de sementes de três espécies do gênero Solanum: S. granuloso-leprosum (gravitinga), S. lycocarpum (lobeira) e S.pseudoquina (quina-de-são paulo). Dependendo da espécie, foram testados diferentes lotes, bem como sementes de diferentes densidades e colorações. Os testes de germinação foram conduzidos a 25ºC e a 20-30ºC, com fotoperíodo de oito horas sob luz branca, sobre papel de filtro. Foram avaliados a porcentagem final e o índice de velocidade de germinação das sementes e determinados os períodos de duração do teste, adotando diferentes critérios de germinação. Os resultados mostraram que as sementes de gravitinga de baixa densidade devem ser descartadas; as sementes de gravitinga de coloração amarela e as de quina-de-são paulo de coloração verde são de maior qualidade fisiológica; houve diferença na germinação das sementes de lobeira e de quina-de-são paulo extraídas de diferentes lotes. A alternância da temperatura e luz beneficiou a germinação das sementes das três espécies. O teste padrão de germinação, teve a duração de 60 dias para as sementes de gravitinga (primeira contagem com 37 dias), de 62 dias para as de quina-de-são paulo (primeira contagem com 33 dias) e de 88 dias para as de lobeira (primeira contagem com 39 dias). Quando o critério botânico para germinação foi adotado (emissão da raiz primária) houve redução no período de duração de germinação em 30 dias para as sementes de gravitinga e de quina-de-são paulo e de 58 dias para as sementes de lobeira.

Metabolic, biochemical and molecular profiling of Catharanthus roseus flower cultivars and transformed hairy roots for monoterpenoid indole alkaloid accumulation /

Catharanthlls rosellS (L.) G Don is a commercially significant flower species and in addition is the only source of the monoterpenoid indole alkaloids (MIA) vinblastine and vincristine, which are key pharmaceutical compounds that are used to combat a number of different cancers. Therefore, procurement of the antineoplastic agents is difficult but essential procedure. Alternatively, CatharanthllS tissue cultures have been investigated as a source of these agents; however they do not produce vindoline, which is an obligate precursor to vinblastine and vincristine. The interest in developing high MIA cultivars of Catharantlws rosellS has prompted metabolic profiling studies to determine the variability of MIA accumulation of existing flowering cultivars, with particular focus on the vindoline component ofthe pathway. Metabolic profiling studies that used high performance liquid chromatography of MIAs from seedlings and young leaf extracts from 50 different flowering cultivars showed that, except for a single low vindoline cultivar (Vinca Mediterranean DP Orchid), they all accumulate similar levels of MIAs. Further enzymatic studies with extracts from young leaves and from developing seedlings showed that the low vindoline cultivar has a IO-fold lower tabersonine-16-hydroxylase activity than those of CatharanthllS rosellS cv Little Delicata. Additionally, studies aimed at metabolic engineering ofvindoline bios}l1thesis in Catharanthus rosellS hairy root cultures have been performed by expressing the last step in vindoline biosynthesis [Dcacetylvindoline-4-0- acetyltransferase (DAT)]. Enzymatic profiling studies with transformed hairy roots have confirmed that over-expressing DAT leads to lines with high levels of O-acetyltransferase activity when compared to non-expressing hairy roots. One particular DA T over111 expressing hairy root culture (line 7) contained 200 times the OAT activity than leaves of control lines. Additional MIA analyses revealed that DAT over-expressing hairy roots have an altered alkaloid profile with significant variation in the accumulation of h6rhammericine. Further analysis of transformed hairy root line 7 suggests a correlation between the expression of OAT activity and h6rhammericine accumulation with root maturation. These studies show that metabolic and selective enzymatic profiling can enhance our ability to search for relevant MIA pathway mutants and that genetic engineering with appropriate pathway genes shows promise as a tool to modify the MIA profile of Catharanthus roseus.

Unique and unifying themes in the mechanisms regulating the expression of the arabidopsis thaliana PR-1 and Solanum tuberosum PR-10a inducible defense genes

Arabidopsis is a model plant used to study disease resistance; Solanum tuberosum or potato is a crop species. Both plants possess inducible defense mechanisms that are deployed upon recognition of pathogen invasion. Transcriptional reprogramming is crucial to the activation of defense responses. The Pathogenesis-Related (PR) genes are activated in these defense programs. Expression of Arabidopsis PR-l and potato PR-10a serve as markers for the deployment of defense responses in these plants. PR-l expression indicates induction of systemic acquired resistance (SAR). Activation of SAR requires accumulation of salicylic acid (SA), in addition to the interaction of the non-expressor of pathogenesis-related genes I (NPRI), with the TGA transcription factors. The PR-10a is activated in response to pathogen invasion, wounding and elicitor treatment. PR-10a induction requires recruitment of the Whirly I (Whyl) activator to the promoter. This locus is also negatively regulated by the silencer element binding factor (SEBF). We established that both the PR-l and PR-10a are occupied by repressors under non-inducing conditions. TGA2 was found to be a constitutive resident and repressor of PR-l, which mediates repression by forming an oligomeric complex on the promoter. The DNA-binding activity of this oligomer required the TGA2 N-terminus (NT). Under resting conditions we determined that the PR-10a is bound by a repressosome containing SEBF and curiously the activator Pto interacting protein 4 (Pti4). In the context of this repressosome, SEBF is responsible for PR-10a binding, yet rWe also showed that PR-l and PR-10a are activated by different means. In PR-l activation the NPRI NT domain alleviates TGA2-mediated repression by interacting with the TGA2 NT. TGA2 remains at the PR-l but adopts a dimeric conformation and forms an enhanceosome with NPRl. In contrast, the PR-10a is activated by evicting the repressosome and recruiting Why! to the promoter. These results advance our understanding of the mechanisms regulating PR-l and PR-10a expression under resting and inducing conditions. This study also revealed that the means of regulation for related genes can differ greatly between model and crop s

Localization of monoterpenoid indole alkaloid (MIA) biosynthesis by in situ hybridization (ISH)

Monoterpenoid indole alkaloids (MIA) are among the largest and most complex group of nitrogen containing secondary metabolites that are characteristic of the Apocynaceae plant family including the most notable Catharanthus roseus. These compounds have demonstrated activity as successful drugs for treating various cancers, neurological disorders and cardiovascular conditions. Due to the low yields of these compounds and high pharmacological value, their biosynthesis is a major topic of study. Previous work highlighting the leaf epidermis and leaf surface as a highly active area in MIA biosynthesis and MIA accumulation has made the epidermis a major focus of this thesis. This thesis provides an in-depth analysis of the valuable technique of RNA in situ hybridization (ISH) and demonstrates the application of the technique to analyze the location of the biosynthetic steps involved in the production of MIAs. The work presented in this thesis demonstrates that most of the MIAs of Eurasian Vinca minor, African Tabernaemontana e/egans and five Amsonia species, including North American Amsonia hubrichitii and Mediterranean A. orienta/is, accumulate in leaf wax exudates, while the rest of the leaf is almost devoid of alkaloids. Biochemical studies on Vinca minor displayed high tryptophan decarboxylase (TOe) enzyme activity and protein expression in the leaf epidermis compared to whole leaves. ISH studies aimed at localizing TOe and strictosidine synthase suggest the upper and lower epidermis of V. minor and T. e/egans as probable significant production sites for MIAs that will accumulate on the leaf surface, however the results don't eliminate the possibility of the involvement of other cell types. The monoterpenoid precursor to all MIAs, secologanin, is produced through the MEP pathway occurring in two cell types, the IPAP cells (Gl0H) and epidermal cells (LAMT and SLS). The work presented in this thesis, localizes a novel enzymatic step, UDPG-7-deoxyloganetic acid glucosyltransferase (UGT8) to the IPAP cells of Catharanthus longifolius. These results enable the suggestion that all steps from Gl0H up to and including UGT8 occur in the IPAP cells of the leaf, making the IPAP cells the main site for the majority of secologanin biosynthesis. It also makes the IPAP cells a likely cell type to begin searching for the gene of the uncharacterized steps between Gl0H and UGT8. It also narrows the compound to be transported from the IPAP cells to either 7-deoxyloganic acid or loganic acid, which aids in the identification of the transportation mechanism.