Qualitative and quantitative study of flavonoids in Notoginseng Radix et Rhizoma based on UPLC-Q-TOF-MS and HPLC-DAD / 中国中药杂志

The flavonoids in Panax notoginseng were qualitatively analyzed by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS), and the content of three main flavonoids in P. notoginseng of different specifications and grades collected from different habitats was determined by HPLC-DAD. Flavonoids and anthocyanins were analyzed by UPLC-Q-TOF-MS/MS in the positive and negative ion modes, respectively. Twelve flavonoid glycosides and one anthocyanin glycoside in P. notoginseng were identified, but no flavonoid aglycones were detected. Among them, 12 compounds were identified in the underground part of P. notoginseng for the first time and eight compounds were first reported in this plant. Moreover, six and four compounds were identified in the Panax genus and the Araliaceae family for the first time, respectively. A method for simultaneous determination of three flavonoids in P. notoginseng was established by HPLC-DAD. The content of flavonoids in 721 P. notoginseng samples of 124 specifications and grades collected from 20 different habitats was simultaneously determined. Among three flavonoids determined, the content of quercetin-3-O-(2″-β-D-xylosyl)-β-D-galactoside was the highest with the average content in the tested samples of 161.0 μg·g~(-1). The content of compounds quercetin-3-O-hexosyl-hexoside and kaempferol-3-O-pentosyl-hexoside was relatively low, with the average content of 18.5 μg·g~(-1)(calculated as quercetin-3-O-sophoroside) and 49.4 μg·g~(-1)(calculated as kaempferol-3-O-sangbu diglycoside). There were significant differences in flavonoids content of samples from different production area. The content of flavonoids in spring P. notoginseng was significantly lower than that in winter P. notoginseng when the other influencing factors such as production areas, germplasm resources, and cultivation conditions were fixed. As for P. notoginseng of different specifications, the flavonoid content in the part connecting the taproot and the aboveground stem was significantly higher than that in other parts. The results of large-scale data showed that the flavonoid content gradually increased with the increase in the number of heads. There were significant differences between the flavonoid content in most specifications and grades, especially the 20-head P. notoginseng and countless head P. notoginseng, whose content was significantly lower and significantly higher than that of other specifications and grades, respectively. This study provides a scientific basis for the study of the effective components and quality control of P. notoginseng from the perspective of flavonoids.

Cyanidin-3-O-glucoside plays a protective role against renal ischemia/ reperfusion injury via the JAK/STAT pathway

Purpose: To investigate the role of cyanidin-3-O-glucoside (C3G) in renal ischemia/reperfusion (I/R) injury and the potential mechanisms. Methods: Mouse models were established by clamping the left renal vessels, and in vitro cellular models were established by hypoxic reoxygenation. Results: Renal dysfunction and tissue structural damage were significantly higher in the I/R group. After treatment with different concentrations of C3G, the levels of renal dysfunction and tissue structural damage decreased at different levels. And its protective effect was most pronounced at 200 mg/kg. The use of C3G reduced apoptosis as well as the expression of endoplasmic reticulum stress (ERS)-related proteins. Hypoxia/reoxygenation (H/R)-induced apoptosis and ERS are dependent on oxidative stress in vitro. In addition, both AG490 and C3G inhibited the activation of JAK/STAT pathway and attenuated oxidative stress, ischemia-induced apoptosis and ERS. Conclusions: The results demonstrated that C3G blocked renal apoptosis and ERS protein expression by preventing reactive oxygen species (ROS) production after I/R via the JAK/STAT pathway, suggesting that C3G may be a potential therapeutic agent for renal I/R injury.

Evaluation of antioxidant activity, phenolic content, anthocyanins, and flavonoids of fresh and dried 'Biloxi' blueberries / Evaluación de la actividad antioxidante, contenido fenólico, antocianinas y flavonoides de arándanos 'Biloxi' frescos y secos

BACKGROUND: The phytochemical content present in blueberries has generated great interest, especially in the nutra-pharmaceutical industry, where it is known as the "super fruit" due to its prevention and treatment of neurodegenerative diseases (cardiovascular diseases, diabetes, and cancer, among others). OBJECTIVES: This study evaluated the functional potential of fresh blueberries and dried blueberries using forced convection by measuring phytochemical content to conclude if this drying technology is convenient for prolonging the product's shelf life. METHODS: For this purpose, antioxidant activity, phenolic content, total anthocyanins, and total flavonoids of 'Biloxi' blueberry cultivars were determined. Fresh and dried blueberries' results were studied. Fruit extracts were analyzed to determine antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) as a free radical, total phenolic content with Folin-Ciocalteu reagent, total anthocyanins by pH differential method, and total flavonoids by Aluminum Chloride method. RESULTS: Results for fresh blueberries yielded ranges of antioxidant activity (90.8-93.9% Free radical scavenging rate), total phenolic content (275 to 645mgGAE/100gFW), total anthocyanins content (28.55 to 43.75mgCy3G/100gFW) and total flavonoids content (159.92 to 335.75mgQE/100gFW). For the forced convection oven process, ranges of antioxidant activity (85.5-92.6% Free radical scavenging rate), total phenolic content (261 to 308mgGAE/100gFW), total anthocyanins content (4.74 to 5.12mgCy3G/100gFW) and total flavonoids content (30.66±0.38mgQE/100gFW) were obtained. CONCLUSIONS: In general, blueberries studied proved to have similar concentrations of functional properties compared to a wide variety of cultivars grown around the globe. Furthermore, higher concentrations of phytochemical content than those reported previously for strawberries, blackberries, and raspberries were evidenced. Although dried blueberries studied proved to have diminished phytochemical content, this functional component content stands out among the fruits market and give nutritional value to end consumers. Drying processes could potentially increase the commerce of blueberries by significantly reducing their perishable nature

CONTEXTO: El contenido fitoquímico presente en los arándanos ha generado gran interés, especialmente en la industria nutra-farmacéutica donde es conocido como una "super fruta" debido a su ayuda en la prevención y tratamiento de enfermedades neurodegenerativas, enfermedades cardiovasculares, diabetes, cáncer, entre otras. OBJETIVOS: Este estudio evaluó el potencial funcional de arándanos frescos y deshidratados por convección forzada mediante la determinación de su contenido fitoquímico con el objetivo de concluir si esta tecnología de secado es conveniente para aumentar la vida útil del producto. MÉTODOS: Para este propósito, se determinó la actividad antioxidante, el contenido fenólico, las antocianinas totales y los flavonoides totales de cultivos de arándanos 'Biloxi' La información recopilada de la literatura fue analizada. Se estudió el contenido en compuestos funcionales en arándanos frescos y deshidratados. Los extractos de fruta fueron analizados para determinar actividad antioxidante por medio de 2,2-Difenil-1-Picrilhidrazilo (DPPH) como radical libre, fenólicos totales con el reactivo Folin-Ciocalteu, antocianinas totales usando el método diferencial de pH y flavonoides totales con el método de Cloruro de Aluminio. RESULTADOS: Para los arándanos frescos se obtuvieron rangos de actividad antioxidante de 90.8-93.9% Tasa de captación de radicales libres, contenido fenólico total de 275-645mgEAG/100gPF, contenido de antocianinas totales de 28.55-43.75mgCy3G/100gPF y contenido total de flavonoides de 159.92-335.75mgEQ/100gPF. Para los arándanos deshidratados por convección forzada, se obtuvieron rangos de actividad antioxidante de 85.5-92.6% Tasa de captación de radicales libres, contenido fenólico total de 261-308mgEAG/100gPF, contenido de antocianinas totales de 4.74-5.12mgCy3G/100gPF y contenido total de flavonoides de 30.24-30.96mgEQ/100gPF. CONCLUSIONES: En general, los arándanos estudiados probaron tener concentraciones similares de propiedades funcionales comparados con una amplia variedad de cultivos alrededor del mundo. Además, fueron evidenciadas concentraciones más altas de contenido fitoquímico comparadas con las reportadas previamente para fresas, moras y frambuesas. Aunque los arándanos secos estudiados demostraron tener menor contenido fitoquímico, la cantidad de estos componentes funcionales destaca dentro del mercado de las frutas y dan valor nutricional a los consumidores. Los procesos de secado pueden potencialmente incrementar el comercio de arándanos derivado de una disminución significativa en su naturaleza perecedera

Efecto de la suplementación del medio de incubación con extracto de Berberis microphylla (calafate) sobre la funcionalidad espermática en semen bovino descongelado / Effect of supplementation of incubation medium with Berberis microphylla (calafate) extract on sperm functionality in thawed bovine semen

RESUMEN: En el semen criopreservado, los procesos de congelación/descongelación y posterior manipulación, dañan las células espermáticas provocando disminución de la capacidad fecundante de los espermatozoides descongelados. Estos procesos han sido asociados con el estado de estrés oxidativo (EO) inducido por altos niveles de especies reactivas de oxígeno (EROS), causando daño a la función y estructura espermática. Los espermatozoides descongelados pueden ser protegidos de este daño, con la adición de antioxidantes (AO) al medio de incubación. El fruto de Calafate (Berberis microphylla G. Forst.) posee una alta capacidad antioxidante, lo que hace interesante investigar el efecto de sus componentes antioxidantes en estos procesos biotecnológicos especialmente postdescongelación. El objetivo de este estudio fue determinar el efecto de la suplementación de extracto liofilizado de fruto de Calafate (ELC), sobre la calidad espermática post-descongelación. Previamente se caracterizó el ELC, determinando la actividad antioxidante y metabolitos como fenoles y antocianinas; posteriormente, espermatozoides de bovino descongelados fueron incubados en un medio base suplementado con diferentes concentraciones de ELC. Post-incubación se evaluó la motilidad progresiva; la viabilidad e integridad de la membrana plasmática (SYBR14- PI) y acrosomal (FITC-PNA/PI) y la peroxidación lipídica (BODIPY) por citometría de flujo. La caracterización de ELC demostró que tanto la actividad antioxidante como los fenoles y antocianinas incrementan concomitante con el aumento de la concentración de ELC. La adición de ELC al medio de incubación, dependiendo de la concentración y tiempo de incubación, sería eficaz en proteger la motilidad, viabilidad e integridad de la membrana plasmática y disminuir la lipoperoxidación en los espermatozoides de bovino descongelados.

SUMMARY: In cryopreserved semen, the freezing/thawing process following of manipulation, damage the sperm cell, decreasing the fertilizing capacity of the thawed sperm; being one of the main factors of this damage the oxidative stress. The sperm once thawed can be protected from this damage, with the addition of antioxidants to the incubation medium. The Calafate fruit (Berberis microphylla G. Forst.) has a high antioxidant capacity, making it an interesting resource for investigating the effect of its antioxidant components on biotechnological processes. The objective of this study was to determine the effect of supplementation of Calafate fruit lyophilized extract (ELC) on sperm quality. The lyophilized extract of the Calafate fruit was characterized, determining the antioxidant activity and metabolites such as phenols and anthocyanins; subsequently, thawed bovine sperm were incubated in a medium supplemented with different concentrations of ELC. Post-incubation, progressive motility was evaluated. By flow cytometry, the viability and integrity of the plasma (SYBR14-PI), and acrosomal (FITC-PNA / PI), as well as lipid peroxidation (BODIPY), was determined. The characterization of Calafate fruits lyophilized extract indicated that antioxidant activity, phenols and anthocyanins increased concomitantly with the increase of dose extract used. The addition of ELC to the incubation medium, depending on the concentration and incubation time, would be effective to protect motility, viability and integrity of the plasma membrane and decreased lipid peroxidation in thawed bovine sperm.

Gall traits and galling insect survival in a multi-enemy context / Características de las agallas, y supervivencia de insectos que producen agallas, en el contexto de enemigos múltiples

Introduction: The enemy hypothesis postulates that gall traits protect galling insects against natural enemies. Galls show a huge range of sizes, colors and ornaments, which vary even intraspecifically. However, galling insects are targets of various organisms that attack them directly or indirectly. In this context, to consider only one gall trait to investigate gall structure acting against only one guild of enemies can conceal an understanding of the community-level interactions. Objective: Herein, we take these ideas into consideration to investigate the conspicuous galls induced by Palaeomystella oligophaga Becker and Adamski 2008 (Lepidoptera) on Macairea radula (Bonpl.) (Melastomataceae) as a model system. We characterize this system through categorization of the different enemy guilds present in the community. We identified them to the lowest taxonomic level possible and determined the kind of interaction responsible for galling insects' deaths. Considering the enemy hypothesis and the selection of secondary characteristics, we also aimed to determine which of the multiple gall traits influence the survival success of galling insects in a multi-enemy context. Methods: We inspected galls and characterized the enemy guilds affecting the galling insect and the mortality rates produced by each one of them. Next, we tested whether the distinct gall traits measured (parenchyma thickness, color, projections) promote galling insect survival with respect to each enemy. Results: The mortality induced by indirect enemies (organisms that interact with gall tissues and can interact secondarily with galling insect) was 47.3 %, being higher than that caused by parasitoids and predators (31.5 %). Despite the gall's structural complexity, live galling insects showed the smallest occurrence (21.2 %). Parenchyma thickness was negatively related to Calliephialtes parasitoids, Gelechiidae cecidophages and predation signals. Conclusions: We demonstrated that the attacks to gall tissues by the cecidophages represented the highest threat to P. oligophaga survival, being higher than the mortality caused by direct enemies. That is, the gall traits were not as efficient as supposed to protect the galling insect from the attack of natural enemies. Nevertheless, we also demonstrated that parenchyma thickness can be negatively related to some organisms, especially direct enemies. Other traits hypothesized as defensive (e.g. projections, coloration) may simply play no role.

Introducción: La hipótesis del enemigo postula que las características de la agalla protegen al agallero contra los enemigos naturales. Las agallas muestran una gran variedad de tamaños, colores y adornos, que varían incluso de forma intraespecífica. Sin embargo, los agalleros son objetivos de varios organismos que los atacan directa o indirectamente. En este contexto, considerar solo una característica para investigar la estructura de la agalla actúando contra un solo gremio de enemigos puede ocultar una comprensión de las interacciones a nivel comunitario. Objetivos: Para investigar las ideas presentadas usamos las agallas conspicuas inducidas por Palaeomystella oligophaga Becker y Adamski 2008 (Lepidoptera) en Macairea radula (Bonpl.) (Meslastomataceae) como sistema modelo. Describimos este sistema a través de la categorización de los diferentes gremios enemigos presentes en la comunidad. Los identificamos al nivel taxonómico más bajo posible y determinamos el tipo de interacción responsable de la muerte de los agalleros. Teniendo en cuenta la hipótesis del enemigo y la selección de características secundarias, también buscamos determinar cuáles de las múltiples caracteristicas de la agalla influyen en el éxito de supervivencia de los agalleros en un contexto de enemigos múltiples. Métodos: Inspeccionamos las agallas y caracterizamos los gremios enemigos que afectan al agallero y las tasas de mortalidad producidas por cada uno de ellos. Luego, probamos si las distintas caracteristicas de las agallas medidas (grosor del parénquima, color, proyecciones) promueven la supervivencia de los agalleros con respecto a cada enemigo. Resultados: La mortalidad indirecta inducida por los cecidofagos fue del 47.3 %, superior a la causada por los parasitoides y los depredadores (31.5 %). Apesar de la complejidad estructural de la agalla, los agalleros vivos mostraron la menor presencia (21.2 %). El grosor del parénquima se relacionó negativamente con los parasitoides de Calliephialtes, los cecidófagos de Gelechiidae y la depredación. Conclusiones: Demostramos que los ataques a los tejidos biliares por cecidófagos representaron la mayor amenaza para la supervivencia de P. oligophaga. Esto es interesante porque, de acuerdo con la hipótesis del enemigo, la estructura de las agallas debería proporcionar protección para los insectos agalleros en lugar de atraer a los insectos que se alimentan de la agalla misma. Sin embargo, también demostramos que el grosor del parénquima puede estar relacionado negativamente con algunos organismos, especialmente los enemigos directos. Otras caracteristicas hipotéticas como defensivas (por ejemplo, proyecciones, coloración) pueden simplemente no desempeñar ningún papel.

Greek yogurt with added sorghum flours: antioxidant potential and sensory acceptance / Yogurt griego adicionado de harinas integrales de sorgo: potencial antioxidante y evaluación sensorial

ABSTRACT The aim of this study was to develop and evaluate the antioxidant characteristics and sensory acceptance of Greek yogurt with added whole sorghum flours (WSF) from genotypes BRS305 (tannin) and BR501 (tannin-free). Five formulations were elaborated: 1) Control (without WSF); 2) 2% BR501; 3) 4% BR501; 4) 2% BRS305 and 5) 4% BRS305 and evaluated for antioxidant capacity, total anthocyanins, total phenols, condensed tannins, sensory acceptance and purchase intention. The 4% BRS305 formulation presented higher antioxidant properties, but lower sensory acceptance. No significant difference was observed for: color, flavor and overall acceptability between the control and 2% BR501; nor for texture and overall acceptability between the control and 4% BR501. Principal component analysis explained that all variables associated with antioxidant properties were positively correlated with the first major component (PC1: 82.7%). The 2% and 4% BR501 formulations were more highly correlated with PC1 (92.7%), as well as the control in relation to sensorial characteristics. The addition of WSF BRS305 improved the antioxidant properties of yogurts and the addition of WSF BR501 did not interfere with the sensory acceptance of the formulations. The similarity of the sensory acceptance of the yogurt containing WSF from the BR 501 genotype to the control, opens perspectives for the insertion of sorghum in human food, using dairy products as suitable matrices, adding potential functionality to this type of product.

RESUMEN El trabajo tuvo como propósito desarrollar y evaluar las características antioxidantes y la aceptación sensorial de yogurt griego con adición de harinas integrales de sorgo (HIS) de los genotipos BRS305 (con taninos) y BR501 (sin taninos). Cinco formulaciones fueron elaboradas: 1) Control (sin HIS); 2) 2% BR501; 3) 4% BR501; 4) 2% BRS305 y 5) 4% BRS305. Las formulaciones fueron evaluadas en función de su actividad antioxidante, antocianinas totales, fenólicos totales, taninos condensados, aceptación sensorial e intención de compra. La formulación 4% BRS305 presentó más propiedades antioxidantes, pero menor aceptación sensorial que las otras formulaciones. No se observó diferencia significativa entre el control y el 2% BR501 para el color, sabor e impresión global, ni tampoco para textura e impresión global en relación al 4% BR501. El análisis de componente principal ha explicado que todas las variables asociadas a las propiedades antioxidantes han sido positivamente correlacionadas con el primer componente principal (PC1: 82,7%). Las formulaciones 2% y 4% BR501 se han correlacionado más con el PC1 (92,7%), así como el control con las características sensoriales. La adición de HIS BRS305 ha mejorado las propiedades antioxidantes de los yogurts y la adición de HIS BR501 no ha interferido en la aceptación sensorial de las formulaciones. La similitud de la aceptación sensorial del yogurt que contiene el HIS del genotipo BR501 con el control, abre perspectivas para la inserción del sorgo en la alimentación humana, utilizando productos lácteos como matrices adecuadas, agregando funcionalidad potencial a este tipo de producto.

Análise de antocianinas e carotenoides em marapuama (Ptychopetalum olacoides) liofilizada / Analysis of anthocyanins and carotenoids in lyophilized marapuama (Ptychopetalum olacoides)

A marapuama é uma planta medicinal com propriedades de interesse em pesquisa. Liofilizar está planta auxilia na preservação de seus compostos. Este trabalho objetivou determinar o conteúdo de antocianinas monoméricas totais e carotenoides totais presentes no liofilizado de marapuama e otimizar a extração. Aplicou-se um DCCR para antocianinas e um para carotenoides. A maior quantidade de antocianina obtida foi de 0,107 mg/100g, e ajustou-se a um modelo onde os termos quadráticos para concentração de etanol e pH foram significativos (p<0,05). Para carotenoides nenhuma das variáveis foi significativa, podendo-se, portanto, usar os menores níveis do planejamento para reduzir custos. A maior quantidade carotenoide foi de 44,21 µg/mL. Conclui-se que quantidades relevantes de compostos antioxidantes foram encontradas em marapuama liofilizada.

Coloração, antocianinas e ácido ascórbico da pitaya colhida em diferentes estágios de maturação / Coloration, antocyanins and ascorbic acid of pitaya collected in different maturation stages

Objetivou-se avaliar a coloração, antocianinas e ácido ascórbico da pitaya vermelha colhida em diferentes estágios de maturação. Os frutos foram oriundos de produtor de Janaúba, MG, colhidos em 2018. O estudo ocorreu em duas etapas: Na primeira etapa, os frutos foram colhidos maduros, no ponto de colheita adotado por produtores locais, sendo avaliados, no mesmo dia da colheita, quanto à coloração da casca (L, C e ⁰Hue), antocianinas totais e ácido ascórbico. Na segunda etapa, foram colhidos comercialmente imaturos e, no 5º dia de armazenamento, passaram pelas mesmas avaliações da etapa 1. Aplicou-se o teste F a 5% de significância. Pitaya colhida em estágio comercialmente imatura apresentou teor de antocianinas totais mais acentuado que na madura. A cor instrumental e o ácido ascórbico apresentaram-se semelhantes ao fruto colhido maduro.

Estabilidade de antocianinas e da cor em polpa de bacaba conservada por métodos combinados / Stability of the anthocyanins and the color bacaba pulp stored by combined methods

O presente estudo teve como objetivo verificar a estabilidade das antocianinas e da cor de polpa de bacaba (Oenocarpus distichus Mart) durante o armazenamento a 25ºC em função de tratamentos combinados: pasteurização (85°C / 3min.), acidificação com acido cítrico (pH 3,7), adição de sacarose (0, 15 e 30% p/p) e adição de conservantes químicos (sorbato de potássio (0, 0,05 e 0,1% p/p), e metabissulfito de sódio (0, 0,01 e 0,02% p/p)). Um planejamento experimental completo do tipo fatorial 32 foi utilizado na elaboração das formulações, que foram armazenadas em BOD, a 25°C, durante 4 meses, sendo que a cada 30 dias eram determinadas as características colorimétricas e o teor de antocianinas. Dos resultados observou-se que, dentro do tempo de armazenamento estudado o teor de antocianinas decresceu após o primeiro mês de armazenamento, com relação a variações de cor, foi observada uma diminuição de a*, H* e C* possivelmente em razão da perda de pigmentos.

Análise sensorial e avaliação da atividade antioxidante de iogurte adicionado de polpa de juçara para valoração do fruto / Sensorial analysis and evaluation of antioxidant activity of yogurt added of jussara pulp for valuation of the fruit

Estudos sobre compostos bioativos com propriedades antioxidantes apontam para benefícios à saúde através de seu uso. Inseridas nesse grupo estão as antocianinas, cujos efeitos se relacionam à redução do estresse oxidativo e dependem da absorção e interação com a microbiota humana. Um ensaio com sistema modelo de iogurte observou aumento da bioacessibilidade e da capacidade antioxidante das antocianinas presentes na juçara quando a fruta participa do processo fermentativo. O presente estudo avaliou a capacidade antioxidante do iogurte adicionado de polpa de juçara. Dessa forma, o iogurte adicionado de polpa foi submetido à quantificação e identificação de antocianinas, determinação da capacidade antioxidante e análise sensorial. Verificou-se que o iogurte desse estudo possui as potencialidades dos compostos presentes na juçara.

Efeito de extratos ricos em antocianinas ou elagitaninos de amora silvestre (Morus nigra L. ), amora preta (Rubus spp), e grumixama (Eugenia brasiliensis Lam) no crescimento e na expressão de genes e miRNAs de diferentes linhagens de céluas humanas de câncer de mama / Effects of extracts rich in anthocyanins or ellagitans from mulberry (Morus nigra L. ), blackberry (Rubus spp) and grumixama (Eugenia brasiliensis Lam) on cellular growth and gene and miRNAs expression from distinctive human breast cancer cell lines

O câncer de mama caracteriza-se globalmente como a neoplasia de maior incidência e mortalidade na população feminina. Antocianinas e elagitaninos presentes em frutas como as berries destacam-se por seu promissor efeito protetor em diferentes estágios do desenvolvimento do câncer de mama. Grumixama (G; Eugenia brasiliensis Lam) é uma espécie de cereja nativa do Brasil que assim como as amora-preta (AP; Rubus spp) e silvestre (AS; Morus nigra L.) contém alto teor de antocianinas e elagitaninos. Poucos estudos focaram na ação anticâncer destas berries no câncer de mama. Portanto, o objetivo do presente estudo foi avaliar os efeitos de extratos ricos em antocianinas ou elagitaninos de G, AP e AS no crescimento e na expressão de genes e miRNAs das linhagens de células humanas de câncer de mama MCF-7 (receptor de hormônio positiva) e MDA-MB 231 (receptor de hormônio negativa). Não se observou citotoxicidade após 72 e 96 horas de tratamento com os extratos (25-200µg/mL) ricos em antocianinas (ASANT, APANT e GANT) ou elagitaninos (APELA e GELA), em ambas as linhagens celulares. Após 72 horas de tratamento, GANT e APANT induziram parada de ciclo celular em G0/G1 (12,5 e 50µg/mL, p<0,05) em células MCF-7. Após 96h, ASANT, APANT e GANT induziram parada de ciclo celular em G0/G1 (12,5 e 50µg/mL; p<0,05) nessas mesmas células. Entretanto, na concentração de 200µg/mL apenas GANT induziu parada em G0/G1 (72 e 96h; p<0,05). Em células MDA-MB 231, após 96h APANT e GANT induziram parada de ciclo celular em G0/G1 nas concentrações testadas (12,5, 50 e 200µg/mL, p<0,05), assim como ASANT nas concentrações de 12,5 e 50µg/mL (p<0,05). Em células MCF-7, após 72h APELA e GELA induziram aumento da proporção de células em subG0 (200µg/mL, p<0,05). Em MDA-MB 231, após 72 e 96h, APELA e GELA (200µg/mL) induziram aumento da proporção de células em subG0 (p<0,05) e parada em G0/G1 (p<0,05). Em células MCF-7, GANT induziu morte celular por apoptose (p<0,05) após 72 e 96h de tratamento. Entretanto, em MDA-MB 231 os extratos ricos em antocianinas não induziram morte celular. Em células MCF-7, após 96h GELA e APELA induziram principalmente necrose (p<0,05). Em MDA-MB 231, APELA e GELA induziram apoptose (p<0,05) após 72 e 96h. Em células MDA-MB 231, após 72h de tratamento foi observada inibição da proliferação celular por GELA, GANT e APELA (200µg/mL; p<0,05). Em células MDA-MB 231, 48h de tratamento com GELA; GANT e APELA (200µg/mL) aumentaram a expressão 5 genes (ESR2, FOXA1, JUN, PTGS2,VEGFA) e inibiram a expressão de 10 genes (ADAM23, ATM, BCL2, CDH1, EGF, GLI1, ID1, MKI67, SNAI2 e THBS1) correlacionados ao câncer de mama. Adicionalmente, GELA; GANT e APELA (200µg/mL) induziram aumento da expressão de miR- 210(p<0,05) e APELA (200µg/mL) reduziu a expressão de miRNA 19a/b (p<0,05) em células MDAMB 231. Coletivamente estes resultados sugerem que antocianinas de grumixama e elagitaninos de amora preta e grumixama apresentam potencial efeito protetor contra o câncer de mama. Adicionalmente, essa ação anticarcinogênica pode ser mediada por indução de morte celular, mais especificamente apoptose, redução de proliferação celular e modulação da expressão de genes e miRNAs relacionados ao câncer de mama.

Breast cancer is characterized as the neoplasia with the highest incidence and mortality rates in women worldwide. Anthocyanins and ellagitannins present in certain fruits, such as berries, stand out for their promising protective effect at different stages of breast cancer development. Grumixama (G; Eugenia brasiliensis Lam), a cherry species from Brazil, as well as blackberry (AP; Rubus spp) and mulberry (AS; Morus nigra L.) contain elevated concentrations of anthocyanins and ellagitannins. Few studies focused on the anticarcinogenic action of these berries in breast cancer development. Therefore, the aim of the present study was to evaluate the effects of extracts rich in anthocyanins or ellagitannins from G, AP and AS on cellular growth and genes and miRNAs expression in human MCF-7 (hormone receptor positive) and MDA-MB 231 (hormone receptor negative) cell lines. No cytotoxicity was observed after 72 and 96 hours of treatment with extracts rich in anthocyanin (25-200 µg/mL) (ASANT, APANT and GANT) or ellagitannins (APELA and GELA) in both cell lines. After 72 hours of treatment, GANT and APANT induced cell cycle arrest at G0/G1 (12.5µg/mL and 50µg/mL, p<0.05) in MCF-7 cells. After 96h, ASANT, APANT and GANT induced cycle arrest at G0/G1 (12.5 and 50µg/mL; p<0.05) in MCF-7 cells. However, at 200µg/mL, only GANT induced G0/G1 (72 and 96h; p<0.05). In MDA-MB 231 cells, after 96h APANT and GANT induced cell cycle arrest at G0/G1 with the three tested concentrations (12.5, 50 and 200µg/mL, p<0.05), as well as ASANT at concentrations 12,5 and 50µg/mL (p <0.05). In MCF-7 cells, after 72h APELA and GELA induced an increase in the proportion of cells in subG0 (200µg/mL, p<0.05). In MDA-MB 231, after 72 and 96h, APELA and GELA (200µg/mL) induced an increase in the proportion of cells in subG0 (p<0.05) and cell cycle arrest in G0/G1 (p<0.05). In MCF-7 cells, GANT induced apoptosis (p<0.05) after 72 and 96h of treatment. However, in MDA-MB 231, extracts rich in anthocyanins did not induce cell death. In MCF-7 cells, after 96h GELA and APELA induced mainly necrosis (p<0.05). In MDA-MB 231, APELA and GELA induced apoptosis (p<0.05) after 72 and 96h. In MDA-MB 231 cells, inhibition of cell proliferation by GELA, GANT and APELA (200?g/mL; p<0.05) was observed after 72h of treatment. In MDA-MB 231 cells, treatment for 48h with GELA, GANT and APELA (200µg) increased expression of 5 genes (ESR2, FOXA1, JUN, PTGS2, VEGFA) and inhibited expression of 10 genes (ADAM23, ATM, BCL2, CDH1, EGF, GLI1, ID1, MKI67, SNAI2 and THBS1) correlated with breast cancer. In addition, GELA; GANT and APELA (200µg/mL) induced increased expression of miR-210 (p<0.05) and APELA (200µg/mL) reduced the expression of miRNA 19a/b (p<0.05) in MDA-MB cells 231. Collectively these results suggest that anthocyanins of grumixama and ellagitannins of blackberry and grumixama have potential protective effect against breast cancer. Additionally, this anticarcinogenic action can be mediated by induction of cell death, more specifically apoptosis, reduction of cell proliferation and modulation of the expression of genes and miRNAs related to breast cancer.

Seasonal behavior of Thalassia testudinum (Hydrocharitaceae) metabolites / Comportamiento estacional de los metabolitos de Thalassia testudinum (Hydrocharital: Hydrocharitaceae)

Abstract:The marine angiosperm Thalassia testudinum, commonly known as turtle grass, is a dominant sea- grass that grows in the Caribbean Sea shelf associated to Syringodium filiforme. The hydroalcoholic extract of T. testudinum is rich in polyphenols; the most abundant metabolite in this extract is thalassiolin B, a glycosilated flavonoid with skin damage repairing properties, and antioxidant capacity among others. The present study aimed at generating information about the seasonal behavior of secondary metabolites, as well as to study the antioxidant capacity of the T. testudinum leaves extract, collected monthly during 2012 from the Northeast coastline of Havana, Cuba. For this study, spectrophotometric methods were used to determine the concentrations of polyphenols, flavonoids, anthocyanins, soluble carbohydrates and proteins, chlorophylls a and b, and antioxidant activity of the extracts. In general, results demonstrated seasonal variations of the analyzed parameters. Extracts prepared from the vegetal material collected in October and November showed the highest values of polyphenols (58.81 ± 1.53 and 52.39 ± 0.63 mg/g bs, respectivally) and flavonoids (44.12 ± 1.30 and 51.30 ± 0.67 mg/g dw, respectively). On the contrary, the lowest values of polyphenols were found in extracts of leaves collected in July and August (15.51 ± 0.84 and 13.86 ± 0.48 mg/g, respectively). In accordance with these results, the lower value of Inhibitory Concentration (IC50) was obtained to get a 50 % of maximal effect on free radical scavenging activity with the extracts prepared from leaves collected in October and November, and less significant IC50 was obtained from the extract prepared from leaves collected in August (5.63 mg/mL). A negative correlation (r= -0.694) was observed in this study between the content of polyphenols and the IC50 necessary to get the half of its antioxidant maximal effect. The high correspondence between the maximum values of polyphenols, flavonoids, carbohydrates and proteins in October and November, revealed a close relationship between these metabolites found in the extract of T. testudinum. Our hypothesis about the annual variation in the concentration of these metabolites was validated; and these results will support the correct harvesting of T. testudinum leaves for biotechnology and industrial purposes. Rev. Biol. Trop. 64 (4): 1527-1535. Epub 2016 December 01.

Resumen:La angiosperma marina Thalassia testudinum, comúnmente conocida como "hierba tortuga", es un pasto marino dominante que crece en el Mar Caribe asociada a Syringodium filiforme. El extracto hidroalcohólico de T. testudinum es rico en polifenoles; el metabolito más abundante en este extracto es thalassiolina B, un flavonoide glicosilado con propiedades para la reparación de daños en la piel y la capacidad antioxidante, entre otros. El objetivo del presente estudio fue conocer el comportamiento estacional de los metabolitos secundarios, así como la capacidad antioxidante del extracto de hojas de T. testudinum recolectadas mensualmente durante el 2012, en la costa noreste de La Habana, Cuba. Para este estudio fueron empleados métodos espectrofotométricos para la determinación de la concentración de polifenoles, flavonoides, carbohidratos y proteínas solubles, clorofilas a y b y la actividad antioxidante del extracto. En sentido general, los resultados demostraron las variaciones estacionales de los parámetros analizados; los extractos preparados a partir de material vegetal recolectado en octubre y noviembre mostraron los mayores valores de polifenoles y flavonoides (44.12 ± 1.30 y 51.30 ± 0.67 mg/g bs respectivamente) y se encontraron los valores más bajos en los extractos de hojas recolectadas en julio y agosto. (15.51 ± 0.84 y 13.86 ± 0.48 mg/g respectivamente). De acuerdo con los resultados, se obtuvo el valor más bajo de la concentración inhibitoria (CI50 ) necesaria para obtener un 50 % de efecto máximo en la actividad de captación de radicales libres con los extractos preparados a partir de las hojas recolectadas en octubre y noviembre y la CI50 menos significativa se obtuvo a partir del extracto preparado a partir de las hojas recolectadas en agosto (5.63 mg/ml). Se observó una correlación negativa (r= -0,694) entre el contenido de polifenoles y la CI50 necesaria para obtener la mitad del efecto antioxidante máximo. La alta correspondencia que existe entre los valores máximos de polifenoles, flavonoides, carbohidratos y proteínas en octubre y noviembre reveló una estrecha relación entre estos metabolitos, encontrados en el extracto de T. testudinum. La hipótesis de la variación anual de la concentración de estos metabolitos fue validada. Estos resultados se tendrán en cuenta con el fin de seleccionar el momento de recolecta de las hojas de T. testudinum para su uso con fines biotecnológicos e industriales.

A comparative proteomic study of white and black glutinous rice leaves

Background Black glutinous rice contains remarkable levels of anthocyanins, which possess anti-oxidative properties and thus have health benefits. The accumulation of anthocyanins in grains of thirty black glutinous rice varieties was measured, and the results revealed that the accumulated anthocyanin content ranged from 0.262 to 2.539 mg/g. Black glutinous rice Br no. 19 was selected, and its leaf protein expression profile was compared with that of white glutinous rice RD 6 using 2D-PAGE, and the protein spots were then directly analyzed after proteolysis by LC-MS/MS. Results The proteins from the leaves of the two rice varieties were separated using 2D-PAGE and silver stained, and the spots were analyzed using Image Master 2D Platinum version 5 software. The results showed that the protein profiles of these two rice cultivars were different, with at least six protein spots that were detected only in Br no. 19. In addition, seven protein spots accumulated at higher levels in Br no. 19 than in RD 6. Conclusion The protein spot S1 (AP005098.4) is homologous to the Rc protein. Our results suggest that some of the proteins enriched in Br no. 19 may be involved in anthocyanin synthesis in the black glutinous rice.

Morphometric and phytochemical characterization of chaura fruits (Gaultheria pumila): a native Chilean berry with commercial potential

BACKGROUND: For the first time, a morphometric characterization of chaura (Gaultheria pumila) fruits has been conducted between natural populations growing in the Villarrica National Park, Araucania Region, Chile. Chaura is a native Ericaceae from Chile that produces aromatic and tasty fruits which could be of agricultural interest. RESULTS: To influence the decision for a further domestication of G. pumila, both the fruit sizes (indicator of productivity) and the nutritional properties of the fruits have been determined from different subpopulations. Samples were a total of 74 plants and 15 fruits per plant which were randomly harvested following its natural distribution around the Villarrica volcano. Altogether, fresh weight, shape, color, diameter in the pole and the equatorial dimensions were determined as phenotypic traits of the G. pumila fruits. Meanwhile the total soluble solids, anthocyanin and pectin contents were calculated as nutritional traits of the Chaura fruits. Results showed a high phenotypic diversity between the sampled population with three main fruit shapes and three predominant colors. The round shapes were the most abundant, whereas a significant correlation was found among fruit size with weight and color. The highest fresh weight (597.3 mg), pole diameter (7.1 mm) and equatorial diameter (6.5 mm) were estimated in the pink color fruits. CONCLUSIONS: The total amount of anthocyanin was higher in red fruits, while the maximum pectin content was obtained in the round white fruits. Overall results must pave the way for a further domestication and introduction of the Chaura species in the agro-productive system in Chile.

Actividad antioxidante de cinco variedades de maíz cultivadas en Campeche, México / Activity antioxidant of maize extracts from Campeche, Mexico

We studied the antioxidant capacity of five varieties of corn grown in Hopelchén, Mexico by the techniques of DPPH, DMPD, oxidation rate, ferric ion reduction and peroxide, the purple variety had the highest antioxidant activity except in test DMPD the red variety which had a better capacity to reduce, in general, the white varieties (native and hybrid) showed similar activity and yellow corn had the lowest activity of all. It also determines the concentration of phenolic compounds and anthocyanins that are present in the corn kernels in a range of 3.39 to 1558 mg of polyphenols and 0,847 to 410 mg of anthocyanins per 100 g of flour. The content of antioxidants in maize varieties can consider it as a functional food by providing exogenous antioxidants to consumers with a consequent protective effects.

Se estudió la capacidad antioxidante de cinco variedades de maíz cultivadas en Hopelchén, México por las técnicas de DPPH (2,2-difenil-1-pricrilhidrazilo), DMPD (N, N,- dimetil-p-fenilendiamina), índice de oxidación, reducción del ion férrico y del peróxido. La variedad morada presentó la mayor actividad antioxidante; excepto en el ensayo de DMPD, en el cual la variedad roja tuvo una mejor capacidad reductora. En general, las variedades blancas (criolla e híbrida) mostraron una actividad similar y, la variedad amarillo tuvo la menor actividad de todas. También se determinó la concentración de compuestos fenólicos y antociánicos que están presentes en las diferentes variedades de maíz en un rango de 3.39 a 1558 mg de polifenoles y de 0.847 a 410 mg de antocianidinas por cada 100 g de harina. El contenido de antioxidantes en las variedades de maíz permite considerarlo como alimento funcional al aportar antioxidantes exógenos a su consumidor con sus consecuentes efectos protectores.

Time-dependence of Ferric Reducing Antioxidant Power (FRAP) index in Chilean apples and berries / Influencia del tiempo de incubación sobre la capacidad antioxidante medida según FRAP, en manzanas y berries chilenos

We hypothesize that the Ferric Reducing Antioxidant Power (FRAP) assay that follows the reaction of Fe3+-TPTZ at 593 nm underestimates the antioxidant capacity of fruits, since the standardized time of the reaction (4 min) is not enough to titrate all the reducing compounds available. We measured FRAP, total phenolics and anthocyanins content in a variety of Chilean berry fruits (blueberries, blackberries, raspberries and strawberries) and apples (cv. Fuji, Granny Smith, Pink Lady, Red Delicious and Royal Gala). Taking into account the dependence of FRAP on the time course of the reaction, we propose to measure FRAP indexes after 1 min (FRAP-1), 30 min (FRAP-30) and 120 min (FRAP-120) of incubation. Most fruit extracts showed significant correlations between the antioxidant capacity and the incubation time, although in some cases the FRAP indexes did not correlate with the total phenolics and/or anthocyanins content. In fact, in apples and berries the correlation between anthocyanins content and FRAP indexes decreased with the incubation time. It is concluded that the fruit extracts analyzed require an incubation period higher than the established in the original experimental protocol to reach the equilibrium, due to the presence of a complex mixture of antioxidant compounds. In addition, a kinetic profile should be realized in each sample studied to establish the most suitable incubation period to titrate all the reactive antioxidant species.

Se plantea que el ensayo de la capacidad antioxidante de frutas, medido según el poder reductor de hierro (FRAP), que sigue la reacción de Fe3+-TPTZ a 593 nm, subestima la capacidad antioxidante, debido a que el tiempo de reacción (4 min) no sería suficiente para que reaccionen todos los compuestos reductores disponibles en las muestras. Se analizó la capacidad antioxidante FRAP, el contenido de fenoles y de antocianinas en diversos berries (arándano, mora, frambuesa y frutilla) y manzanas (cv. Fuji, Granny Smith, Pink Lady, Red Delicious y Royal Gala). Tomando en cuenta la dependencia del tiempo de incubación en el valor FRAP, se propone medir los índices FRAP después de 1 min (FRAP-1), 30 min (FRAP-30) y 120 min (FRAP-120). Diversos extractos de las frutas analizadas mostraron una correlación significativa entre la capacidad antioxidante y el tiempo de incubación; sin embargo, en algunos casos los índices FRAP no se correlacionaron con el contenido de fenoles totales y/o antocianinas. En efecto, en manzanas y berries la correlación entre el contenido de antocianinas e índices FRAP disminuyó con el tiempo de incubación. Se concluye que los extractos analizados requieren un tiempo de incubación mayor al que establece el protocolo analítico original para alcanzar el equilibrio, debido a la presencia de una compleja mezcla de compuestos antioxidantes. Además, el perfil cinético de cada muestra debería ser estudiado para establecer el periodo de incubación más adecuado para titular todas las especies antioxidantes reactivas.

Correlação entre atividade antioxidante e compostos bioativos de polpas comerciais de açaí (Euterpe oleracea Mart)

Este trabalho objetivou caracterizar polpas comerciais de açaí, além de correlacionar a contribuição da vitamina C, carotenóides totais e compostos fenólicos com a atividade antioxidante desses produtos, verificando ainda os parâmetros referentes à qualidade dos produtos obtidos. Foram realizadas análises de pH, acidez total titulável, sólidos solúveis totais, açúcares, cor, atividade de água, vitamina C, antocianinas totais e carotenóides totais, atividade antioxidante e compostos fenólicos. Conclui-se que todos os parâmetros apresentaram diferença significativa entre as marcas não adicionadas de conservante e sacarose. O açaí pode ser considerado uma importante fonte de vitamina C, bem como uma grande fonte de antioxidantes. Em relação à correlação com a atividade antioxidante, somente as antocianinas totais e os carotenóides totais apresentaram correlação positiva e significativa.

Correlation between antioxidant activity and bioactive compounds of açaí (Euterpe oleracea Mart) comercial pulps. The purpose of this work was to analyze commercial açaí pulp and to correlate the contribution of vitamin C, total carotenoids and phenolic compounds with the antioxidant activity of there products besides the examination of the standard related to the quality of the obtained products. Analysis were made with regard to pH, acidity total, soluble solids, sugars, color, water activity, vitamin C, total anthocyanins, total carotenoids, antioxidant activity and phenolic compounds. All parameters analyzed showed significative difference among the marks not added with preservatives and sucrose. The açaí fruit can be considered a fair source of vitamin C and good source of natural antioxidants. In regard to the correlation with the antioxidant activity only the total anthocyanins and the total carotenoids presented positive and significative correlation.

Caracterización del acai o manaca (Euterpe olerdcea Mart. ): un fruto del Amazonas / Characterization of the acai or manaca (Euterpe oleracea Mart. ): a fruit of the Amazon

The objective of this study was to determine the proximal composition, the fatty acid profile, the content of minerals, tannins, polyphenols, anthocyanins, the antioxidant capacity and the color of the acai pulp (Euterpe oleracea Mart) collected in the Venezuelan Amazon from 2 harvests of the year 2005. For the proximal analysis, the official methods were used and the minerals were analyzed by the induced plasma technique. Polyphenols, tannins and anthocyanins were determined by spectrophotometric methods and the antioxidant capacity was analyzed by DPPH method. Results in dry basis indicated that acai has a high lipids content (49.4% and 33.1%), proteins (13.8% and 9.3%), ash (5.2% and 2.2%) and total dietary fiber (27.3% and 18.0%). It stands out that 71% of the acai fat is oleic acid and that the Fe content of the first and second harvest was 0.023 and 0.015 mg/100g, respectively; polyphenols 5.02 and 2.20 g/100 g; tannins 0.70 and 1.37 g/100g; anthocyanins 0.73 and 1.60 g/100g and the antioxidant capacity 88.03 and 87.87%, respectively. It is concluded that the acai or manaca collected in the Venezuelan Amazon has a high nutritional value and contains antioxidant compounds which suggests the need to industrialize it to take advantage to the maximum of its properties.

Phenotypic plasticity in Passiflora suberosa L.(Passifloraceae): induction and reversion of two morphs by variation in light intensity / Plasticidade fenotípica em Passiflora suberosa L. (Passifloraceae): indução e reversão de duas formas pela variação na intensidade luminosa

Leaf morphology may vary considerably even within a branch of Passiflora suberosa plants. Leaves are of a typical green type in shaded areas, but in open fields turn into violet, and apparently have greater thickness and trichome density. The proximate causes and the adaptive meaning, if any, for the existence of the violet morph are still unknown. By cultivating P. suberosa clones under two light regimes (total and partial exposure to sunlight), we consecutively induced (first year) and then reversed (second year) the appearance of the violet morph. We evaluated the corresponding changes in morpho-anatomic and chemical leaf characteristics. Plants that were grown under partial sunlight had a greater size and did not alter their green color, but those grown under total sunlight changed into violet, were smaller in size and their leaves were tougher, thicker, and had a greater number of trichomes. The violet morph had increased anthocyanins and phenolic derivatives. It also showed cellular hypertrophy, a greater number of cell layers in the mesophyll, and a lignified pericycle. Since these morphs are interchangeable by changing light conditions, we inferred that they are not determined by genotypic diversity, but are mainly a result of a physiological response to light stress, and thus part of P. suberosa phenotypic plasticity.

A morfologia das folhas de Passiflora suberosa pode variar consideravelmente mesmo dentro dos ramos de um dado espécime. P. suberosa ocorre tipicamente em áreas sombreadas e as folhas são verdes. Porém, em áreas abertas, onde há maior incidência de luz solar, as folhas são de coloração roxa, aparentemente mais duras e com grande densidade de tricomas. As possíveis causas e o significado adaptativo da manifestação destas características ainda são desconhecidas. Com base no cultivo de clones de P. suberosa sob dois regimes de luz solar (incidência total e parcial), nós consecutivamente induzimos (primeiro ano) e então revertemos (segundo ano) o aparecimento da forma roxa. As mudanças nas características morfológicas e químicas das formas verde e roxa foram avaliadas. As plantas que foram cultivadas sob incidência parcial de luz solar apresentaram maior tamanho dos ramos e não alteraram a cor verde das folhas. As plantas que foram cultivadas sob incidência total dos raios solares apresentaram coloração roxa, maior dureza, espessura e pilosidade. A forma roxa apresentou alto teor de antocianinas e derivados fenólicos. As plantas exibiram hipertrofia celular, maior número de camadas celulares no mesofilo e lignificação do periciclo. Considerando que as formas são intercambiáveis perante a mudança na intensidade luminosa, nós inferimos que elas não resultam da diversidade genotípica, mas sim de uma resposta fisiológica ao estresse luminoso e, dessa forma, parte da plasticidade fenotípica de P. suberosa.

Evoluçäo dos fenólicos totais e taninos condensados (Proantocianidinas) durante o desenvolvimento das sementes do feijäo (Phaseolus vulgaris L.) / Evolution of phenolic compounds and condensed tannins (Proanthocyanidins) during the development of bean seeds (Phaseolus vulgaris L.)

The evolution of phenolic compounds and tannins (proanthocyanidins) of bean seeds, Phaseolus vulgaris L., (cultivar aroana 80), from anthesis to maturity (10 to 45 days after anthesis), was investigated. During seed development, phenolic compounds and tannins contents increased by seed unit, until the 31st and 21st day after anthesis respectively, decreasing afterwards. The gradual decrease in catechin and the increase of its polymers (intermediate compounds), as well as the gradual increase in alpha-amylase inhibition capacity were indications that tannins polymerize during seed development