Impact of phenylalanine on Hanseniaspora vineae aroma metabolism during wine fermentation.

Hanseniaspora vineae exhibits extraordinary positive oenological characteristics contributing to the aroma and texture of wines, especially by its ability to produce great concentrations of benzenoid and phenylpropanoid compounds compared with conventional Saccharomyces yeasts. Consequently, in practice, sequential inoculation of H. vineae and Saccharomyces cerevisiae allows to improve the aromatic quality of wines. In this work, we evaluated the impact on wine aroma produced by increasing the concentration of phenylalanine, the main amino acid precursor of phenylpropanoids and benzenoids. Fermentations were carried out using a Chardonnay grape juice containing 150 mg N/L yeast assimilable nitrogen. Fermentations were performed adding 60 mg/L of phenylalanine without any supplementary addition to the juice. Musts were inoculated sequentially using three different H. vineae strains isolated from Uruguayan vineyards and, after 96 h, S. cerevisiae was inoculated to complete the process. At the end of the fermentation, wine aromas were analysed by both gas chromatography-mass spectrometry and sensory evaluation through a panel of experts. Aromas derived from aromatic amino acids were differentially produced depending on the treatments. Sensory analysis revealed more floral character and greater aromatic complexity when compared with control fermentations without phenylalanine added. Moreover, fermentations performed in synthetic must with pure H. vineae revealed that even tyrosine can be used in absence of phenylalanine, and phenylalanine is not used by this yeast for the synthesis of tyrosine derivatives.

Health-Promoting Potential of Mandarin Pomace Extracts Enriched with Phenolic Compounds.

The aim of this work was to assess the effect of in vitro human digestion on the chemical composition (carbohydrates and phenolic compounds) and bioactivity of hydro-alcoholic-acid pomace extracts from two mandarin varieties (Clemenule and Ortanique) by measuring their antioxidant, antidiabetic, anti-glycative, hypolipidemic, and anti-inflammatory properties. The phenolic compound profile showed that nobiletin was the main flavonoid found in the extracts and digests of Clemenule pomace and extract, while isosinensetin/sinensetin/tangeretin were the ones in the Ortanique samples. The digests of Clemenule and Ortanique extracts showed Folin reaction values of 9.74 and 9.20 mg gallic acid equivalents (GAE)/g of sample, ABTS values of 83.2 and 91.7 µmol Trolox equivalents (TE)/g of sample, and ORAC-FL values of 142.8 and 891.6 µmol TE/g of sample, respectively. Extracts (50-500 µg/mL) inhibited intracellular reactive oxygen species (ROS) formation in CCD-18Co cells under physiological and oxidative-induced conditions. Clemenule and Ortanique extract digests showed IC50 values of 13.50 and 11.07 mg/mL for α-glucosidase, 28.79 and 69.64 mg/mL for α-amylase, and 16.50 and 12.77 mg/mL for AGEs, and 2.259 ± 0.267 and 0.713 ± 0.065 mg/mL for pancreatic lipase inhibition, respectively. Ortanique extract (250-1000 µg/mL) inhibited the production of nitric oxide in RAW264.7 macrophages under inflammation-induced conditions, and intracellular ROS formation. In conclusion, altogether, the results supported the potential of mandarin extracts to be used as health promoters by reducing the risk of non-communicable chronic diseases.

Protección del ácido clorogénico frente a lesiones inducidas por venzo(a)pireno en Saccharomyces cerevisiae / Protection of chlorogenic acid against benzo(a)pyrene-induced lesions in Saccharomyces cerevisiae / Proteção do ácido clorogênico contra lesões induzidas por benzo(a)pireno em Saccharomyces cerevisiae

El objetivo del presente estudio fue analizar el efecto del ácido clorogénico, uno de los compuestos polifenólicos con mayor concentración en la infusión de Ilex paraguariensis, sobre el daño celular y molecular inducido por el benzo(a)pireno. La infusión de Ilex paraguariensis ("mate") es bebida por la mayoría de los habitantes de Argentina, Paraguay, sur de Brasil y Uruguay. La levadura Saccharomyces cerevisiae (cepas SC7K lys2-3; SX46A y SX46Arad14() se utilizó como modelo eucariota. Las células en crecimiento exponencial se expusieron a concentraciones crecientes de benzo(a)pireno y a tratamientos combinados con una concentración de 250 ng/mL de benzo(a)pireno y ácido clorogénico a una concentración igual a la encontrada en la infusión de yerba mate. Luego de los tratamientos se determinaron fracciones de sobrevida, frecuencia mutagénica y roturas de doble cadena de ADN así como la modulación en la expresión de la proteína Rad14 a través de un análisis de Western Blot. Se observó un aumento significativo en las fracciones de sobrevida así como una disminución en la frecuencia mutagénica después de la exposición combinada con benzo(a)pireno y ácido clorogénico en comparación con los tratamientos con benzo(a)pireno como único agente. En la cepa mutante deficiente en la proteína Rad14 se observó un aumento significativo en la sensibilidad a benzo(a)pireno en comparación con la cepa SC7K lys2-3. En los tratamientos combinados de benzo(a)pireno y ácido clorogénico se observó una importante disminución de la letalidad. Con respecto a la determinación de roturas de doble cadena de ADN no se observó fraccionamiento cromosómico a la concentración de benzo(a)pireno utilizada en los experimentos. Los análisis de Western Blot mostraron un aumento en la expresión de la proteína Rad14 en las muestras tratadas con benzo(a)pireno como único agente en comparación con la muestra control. Adicionalmente se observó una disminución en la expresión de la proteína cuando en los tratamientos se utilizaron benzo(a)pireno y ácido clorogénico combinados. Los resultados indican que el ácido clorogénico disminuye significativamente la actividad mutagénica producida por el benzo(a)pireno, la cual no se encuentra relacionada con un incremento en la remoción de las lesiones inducidas por el sistema de reparación por escisión de nucleótidos.

The aim of this study was to analyze the effect of chlorogenic acid, a polyphenolic compound found at high concentrations in Ilex paraguariensis infusions, on cellular and molecular damage induced by benzo(a)pyrene. Ilex paraguariensis infusions ("mate") are consumed by most of the population in Argentina, Paraguay, southern Brazil and Uruguay. Saccharomyces cerevisiae yeast (SC7K lys2-3; SX46A and SX46Arad14( strains) were used as eukaryotic model organisms. Cells in an exponential growth phase were exposed to increasing concentrations of benzo(a)pyrene, as well as combined treatments of benzo(a)pyrene at a concentration of 250 ng/mL and chlorogenic acid at a concentration matching that which is commonly found in mate. Determinations of surviving fraction, mutagenic frequency and double strand DNA breaks induction were performed, along with the assessment of the modulation of the expression of protein Rad14 by Western Blot. A significant increase of surviving fractions and a decrease in mutagenic frequency were observed after exposure to benzo(a)pyrene plus chlorogenic acid, contrary to benzo(a)pyrene alone. A substantial increase in sensitivity to benzo(a)pyrene was observed for the Rad14 protein-deficient mutating strain when compared to the SC7K lys2-3 strain. An important decrease in lethality was observed when combined benzo(a)pyrene and chlorogenic acid treatments were applied. As for the determination of DSBs, no chromosomic fractionation was observed at the benzo(a)pyrene concentration tested in the experiments. Western Blot analysis showed an increase in the expression of protein Rad14 for samples treated with benzo(a)pyrene as a single agent when compared against the control sample. Additionally, the expression of this protein was observed to diminish when combined treatments with benzo(a)pyrene and chlorogenic acid were used. Results obtained indicate that chlorogenic acid significantly decreases the mutagenic activity of benzo(a)pyrene, which is not related to an increase in the removal of lesions induced by nucleotide excision repair system.

O objetivo deste estudo foi analisar o efeito do ácido clorogênico, um dos compostos polifenólicos com maior concentração na infusão de Ilex paraguariensis, sobre o dano celular e molecular induzido pelo benzopireno. A infusão de Ilex paraguariensis ("mate") é consumida pela maioria dos habitantes da Argentina, Paraguai, sul do Brasil e Uruguai. A levedura Saccharomyces cerevisiae (cepas SC7K lys2-3; SX46A e SX46Arad14() foi utilizada como modelo eucariótico. Células em crescimento exponencial foram expostas a concentrações crescentes de benzopireno e tratamentos combinados foram realizados com uma concentração de 250 ng/mL de benzo(a)pireno e ácido clorogênico, igual à encontrada na infusão de erva-mate. Após os tratamentos, foram determinadas as frações de sobrevivência, frequência mutagênica e quebras de fita dupla do DNA, bem como a modulação na expressão da proteína Rad14 por meio de análise de Western Blot. Um aumento significativo nas frações de sobrevivência, bem como uma diminuição na frequência mutagênica foram observados após a exposição combinada de benzo(a)pireno e ácido clorogênico em comparação com tratamentos de agente único de benzo(a)pireno. Um aumento significativo na sensibilidade ao benzo(a)pireno foi observado na cepa mutante deficiente em proteína Rad14 em comparação com a cepa SC7K lys2-3. Nos tratamentos combinados de benzo(a)pireno e ácido clorogênico, observou-se uma diminuição significativa na letalidade. Com relação à determinação das quebras de fita dupla de DNA, não foi observado fracionamento cromossômico na concentração de benzo(a)pireno utilizada nos experimentos. A partir da análise de Western Blot, observou-se um aumento na expressão da proteína Rad14 nas amostras tratadas com benzo(a)pireno como agente único em relação à amostra controle. Além disso, uma diminuição na expressão da proteína foi observada quando combinados de benzo(a)pireno e ácido clorogênico foram usados ​​nos tratamentos. Os resultados obtidos neste trabalho indicam que o ácido clorogênico diminui significativamente a atividade mutagênica produzida pelo benzo(a)pireno, a qual não está relacionada a um aumento na remoção de lesões induzidas pelo sistema de reparo por excisão de nucleotídeos.

Validación del uso tradicional de especies de asclepias contra el veneno de bothrops diporus (yarará chica) en el nordeste de Argentina / Validation of the traditional use of asclepias species against bothrops diporus (yarará chica) venom in northeast of Argentina

La familia Asclepiadaceae posee tradición en etnomedicina. En el nordeste argentino, A.mellodora y A. curassavica se utilizan como cataplasmas en accidentes de ofidios. En este trabajo, los extractos acuosos, etanólicos y hexánicos de A. mellodora y A. curassavica se evaluaron por SDS-PAGE para determinar su actividad alexitérica. El estudio in vitro de la capacidad inhibitoria de las actividades proteolítica, hemolítica indirecta y coagulante permitieron determinar que ambas especies manifiestan actividad, siendo A. mellodora más activa. Estadísticamente los extractos de A. mellodora fueron igualmente activos contra el veneno de Bothrops diporus y no mostraron diferencias significativas respecto del órgano utilizado en la inhibición de la actividad coagulante. Este resultado está en consonancia con la forma tradicional de su uso como cataplasma. Sobre el extracto etanólico de las raíces de A. mellodora se realizó un fraccionamiento bioguiado que permitió identificar fracciones de compuestos responsables de la actividad.

The Aclepiadaceae family has been reported by its use in ethnomedicine. In the northeast of Argentina, A. mellodora and A. curassavica are traditionally used in ofidic accidents as poultices. In this work, aqueous, alcoholic and hexanoic extracts were analyzed by SDS-PAGE to determine their anti-snake activity. The in vitro study of the inhibitory ability of the following activities: proteolytic, indirect hemolytic activity and inhibition of the coagulant activity, allowed demonstrating that both species were active against venom, being A. mellodora the most active. Statistically, all extracts of A. mellodora were active against venom in the inhibition of the coagulant activity, without significant differences with respect to the organ used; which is consistent with the traditional use as external poultice. The alcoholic extract of A. mellodora roots was subjected to a bio-guided separation. The fractions obtained were enriched in compounds which could probably be responsible for the activity against venom.

SDS-PAGE: una herramienta útil en la evaluación preliminar de la actividad alexítera de extractos vegetales / SDS PAGE: a useful tool for preliminary screening of antisnake activity of plant extracts

Some results obtained during our research work in the search of anti-snake compounds from plant origin, allow us to propose sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a valuable method for a fast and reliable screening in order to evaluate plant extracts activity on snake proteins from Bothrops diporus (yarará chica). Such approach will allow to process a larger number of plant extracts and to select the active ones. Venoms used in this study came from B. diporus which was previously vacuum dried. Extracts (aqueous, alcoholic and hexanic) were from native plants: Aristolochia elegans, Aristolochia gibertii, Asclepia curassavica, Cissampelos pareira, Dorstenia brasiliensis, Eclipta prostrata, Iresine diffusa, Mikania micrantha, M. periplocifolia, M. coridifolia, Nectandra angustifolia, N. megapotamica, Sapium haematospermum and Trixis divaricata. The results obtained by SDS-PAGE were compared with those obtained from in vitro assays (coagulation and hemolysis inhibition). The correlation between results obtained from electroforetic and in vitro assays allowed to suggest SDS-PAGE as a suitable technique to assist in preliminary plant screenings for anti-snake activity by snake venom protein interaction with plant compounds.

El desarrollo de nuestro trabajo de investigación en la búsqueda de compuestos alexíteros de origen vegetal nos permite proponer la electroforesis en geles de poliacrilamida en condiciones desnaturalizantes, como método de screening rápido y confiable, para evaluar la actividad de extractos vegetales sobre proteínas del veneno de yarará, de manera de procesar mayor número de muestras vegetales y seleccionar aquellas que son activas. Para el desarrollo de la metodología, se utilizó un pool de veneno de Bothrops diporus desecado al vacío y extractos acuosos, alcohólicos y hexánicos de plantas autóctonas Aristolochia elegans, A. gibertii, Asclepia curassavica, Cissampelos pareira, Dorstenia brasiliensis, Eclipta prostrata, Iresine diffusa, Mikania micrantha, M. periplocifolia, M. coridifolia, Nectandra angustifolia, N. megapotamica, Sapium haematospermum y Trixis divaricata. Se realizaron pruebas in vitro (inhibición de la coagulación y hemólisis) para contrastar con los resultados obtenidos por SDS-PAGE. La correlación de los resultados obtenidos con técnicas in vitro validadas, permite sugerir el empleo de la técnica de SDS-PAGE como una herramienta útil en la evaluación preliminar de la actividad alexítera de extractos vegetales, propiedad evidenciada por la modificación en el perfil de bandas proteicas cuando se compara el veneno puro con el producto de la interacción extracto vegetal-veneno.

Efeito fungicida dos óleos essenciais de Schinus molle L e Schinus terebinthifolius Raddi, Anacardiaceae, do Rio Grande do Sul / Antifungal effect of Schinus molle L, Anacardiaceae, and Schinus terebinthifolius Raddi, Anacardiaceae, essential oils of Rio Grande do Sul

Uma das alternativas para reduzir o uso de agrotóxicos pode ser a utilização de óleos essenciais para o controle de doenças e pragas que afetam a produção agrícola. O objetivo do presente estudo foi verificar o efeito fungicida dos óleos essenciais de Schinus molle L. e Schinus terebinthifolius Raddi, espécies nativas amplamente distribuídas na região. Os óleos foram extraídos por hidrodestilação e analisados por cromatografia gasosa. Os fungos fitopatógenos Alternaria spp., Botrytis spp., Colletotrichum spp. e Fusarium spp., foram isolados a partir de gérberas e rosas com sintomas de doença e os ensaios de atividade antifúngica foram realizados através da técnica de difusão em disco e os dados analisados pelo teste de Tukey e Duncan a 5 por cento. Em geral o óleo essencial de S. molle foi efetivo para os quatro fungos testados, nas quatro diluições utilizadas e para todos os tempos avaliados e o óleo essencial de S. terebinthifolius apresenta efeito fungicida mais pronunciado contra Botrytis spp., a partir de testes in vitro.

One of the alternatives to reduce the use of pesticides may be the use of essential oils to control diseases and pests that affect agricultural production. The purpose of this study was to evaluate the fungicide effect of essential oils of Schinus molle L. and Schinus terebinthifoliusRaddi, native species widely distributed in the region. The oils were extracted by hydro distillation and analyzed by gas chromatography. The phytopathogenic fungi Alternariaspp., Botrytisspp., Colletotrichumspp. and Fusarium spp., were isolated from "gerberas" and roses with symptoms of disease and antifungal activity of the tests were carried out by the technique of spreading disk. The data was analyzed by Tukey and Duncan tests to 5 percent. In general, the essential oil of S. mollewas effective for the four fungi tested, for the four dilutions used and for all the times evaluated. The essential oil of S. terebinthifolius presents more pronounced fungicide effect against Botrytisspp., from in vitro tests.

Plantas Medicinales y Fitoterapia: ¿Indicadores de Dependencia o Factores de Desarrollo? / Medicinal Plants and Phytotherapy: Indicators of Dependence or Development Factors?

Los medicamentos fitoterápicos están constituidos por ingredientes activos de origen vegetal formulados bajo la forma farmacéutica más adecuada para su administración. La posibilidad de utilización de la Fitoterapia en las prácticas terapéuticas, con sustento científico, exige acciones multisectoriales que involucren desde la producción primaria de plantas medicinales hasta el establecimiento de los procesos de control de calidad de las materias primas y medicamentos. La Fitoterapia podría transformarse entonces, de ser actualmente un indicador de dependencia, en un factor real de desarrollo para la agroindustria farmacéutica de Latinoamérica, si fuera encarada por las políticas de ciencia y tecnología regionales y nacionales con un criterio más moderno y ecuánime, y en consonancia con las necesidades de la población. (AU)

"Medicinal Plants and Phytotherapy: Dependence Indicators or Development Issues?". Phytomedicines are constituted exclusively by active principles from plant origin formulated under the best pharmaceutical form adapted for their administration. The application of Phytotherapy in therapeutic practices, with scientific support, demands multisectorial actions that involve from the primary production of medicinal plants to the establishment of quality control processes of raw materials and medicines. Once accomplished these objectives, Phytoterapy could become, from a dependency indicator in a real development factor for the pharmaceutical agroindustry in Latin America, if outside faced by the regional and national policies of science and technology with a more modern and even-tempered criterion, and in agreement with the population needs. (AU)

Essential oils of camphor tree (Cinnamomum camphora Nees & Ebern) cultivated in Southern Brazil

Os óleos essenciais de duas variedades da árvore canforeira (Cinnamomum camphora Nees & Eberm, Lauracea), conhecidas como Hon-Sho e Ho-Sho cultivadas em canteiros experimentais no sul do Brasil foram estudados. Os óleos essenciais foram obtidos das folhas e ramos de plantas jovens por hidrodestilaçäo. A identificaçäo dos componentes foi feita por GC, GC/MS e índices de retençäo nas fases metil silicone e carbowax. Os principais componentes foram linalol no Ho-Sho e cânfora no Hon-Sho