Strawberries treated with biodegradable film containing plant extracts.

Strawberry (Fragaria x ananassa Duch.) is a highly perishable fruit whose characteristics make it susceptible to developing microorganisms. Plant extracts have been studied as an alternative to pesticides to control spoilage microorganisms, responding to the expectation of the population seeking a healthier way of life. The fungus Botrytis cinerea is a facultative pathogen of vegetables, which can affect all stages of the development of several fruits, such as the strawberry, where it causes gray rot. Trichilia catigua (catuaba), Paullinia cupana (guarana), Stryphnodendron barbatiman (barbatimão), and Caesalpinia peltophoroides (sibipiruna) are planted in the Brazilian flora and have demonstrated pharmacological properties in their extracts. This work aimed to treat strawberries with a biodegradable film containing extracts of these species to evaluate strawberry conservation. There were notable distinctions in mass loss between the extract-treated and control samples. The pH, total acidity (TA), and soluble solids parameters exhibited consistently significant means across both sets of samples. Luminosity increased over the course of days in the color parameters, with the exception of strawberries coated with guarana. The red color showed greater intensity, except for those coated with barbatimão extract. Considering the results, it is possible to conclude that the coatings used can become an alternative to enhance the conservation of strawberries.

Cytotoxicity of extracts and compounds isolated from Croton echioides in animal tumor cell (HTC).

The search for compounds with anticancer effects is of paramount importance today due to the high incidence of the disease. The Euphorbiaceae family is known for having compounds with therapeutic properties, one of its genera being Croton. It has several species, which contain compounds already known for their biological activities, presenting anti-inflammatory, antimicrobial and anticancer properties. Thus, the cytotoxicity/antiproliferative activity of semi-purified fractions and compounds isolated from Croton echioides in liver tumor cells of Rattus norvegicus (HTC) was evaluated by the MTT test. The semi-purified fractions showed cytotoxicity at concentrations above 200 µg/mL, at 24, 48 and 72 hours, reaching cell viability of 24.78% [400 µg/mL] at 24 hours, 12.79% [500 µg/mL] at 48 hours and 10.57% [300 µg/mL] at 72 hours. For the isolated compounds, lupeol had a cytotoxic effect in all concentrations (1, 5, 10, 15, 20, 40, 60, 80 and 100 µg/mL) and tested times (24, 48 and 72 hours), reaching minimum viability of 4.37% [100 µg/mL], within 72 hours. The clerodan diterpenes CEH-1 and CEH-4 also showed antiproliferative activity, with minimum viability of 36.19% [100 µg/mL] over 72 hours and 21.33% [100 µg/mL] over 48 hours, respectively. However, the clerodan diterpenes CEH-2 and CEH-3 did not shows a cytotoxic effect for HTC cells. Thus, there is a cytotoxic/antiproliferative potential of C. echioides against tumor cells, with targeted to mitochondrial enzymes, associated with cell proliferation, indicating that this species deserves prominence in the search for new molecules for the treatment of cancer.

Cytotoxicity of extracts and compounds isolated from Croton echioides in animal tumor cell (HTC) / Citotoxicidade de extratos e compostos isolados de Croton echioides em célula tumoral animal (HTC)

The search for compounds with anticancer effects is of paramount importance today due to the high incidence of the disease. The Euphorbiaceae family is known for having compounds with therapeutic properties, one of its genera being Croton. It has several species, which contain compounds already known for their biological activities, presenting anti-inflammatory, antimicrobial and anticancer properties. Thus, the cytotoxicity/antiproliferative activity of semi-purified fractions and compounds isolated from Croton echioides in liver tumor cells of Rattus norvegicus (HTC) was evaluated by the MTT test. The semi-purified fractions showed cytotoxicity at concentrations above 200 µg/mL, at 24, 48 and 72 hours, reaching cell viability of 24.78% [400 µg/mL] at 24 hours, 12.79% [500 µg/mL] at 48 hours and 10.57% [300 µg/mL] at 72 hours. For the isolated compounds, lupeol had a cytotoxic effect in all concentrations (1, 5, 10, 15, 20, 40, 60, 80 and 100 µg/mL) and tested times (24, 48 and 72 hours), reaching minimum viability of 4.37% [100 µg/mL], within 72 hours. The clerodan diterpenes CEH-1 and CEH-4 also showed antiproliferative activity, with minimum viability of 36.19% [100 µg/mL] over 72 hours and 21.33% [100 µg/mL] over 48 hours, respectively. However, the clerodan diterpenes CEH-2 and CEH-3 did not shows a cytotoxic effect for HTC cells. Thus, there is a cytotoxic/antiproliferative potential of C. echioides against tumor cells, with targeted to mitochondrial enzymes, associated with cell proliferation, indicating that this species deserves prominence in the search for new molecules for the treatment of cancer.

A busca por compostos com efeitos anticâncer é de suma importância nos dias atuais devido à alta incidência desta doença. A família Euphorbiaceae é conhecida por possuir compostos com propriedades terapêuticas, sendo um de seus gêneros o Croton. Este possui diversas espécies, que contêm compostos já conhecidos por suas atividades biológicas, apresentando propriedades anti-inflamatórias, antimicrobianas e anticancerígenas. Assim, a citotoxicidade/atividade antiproliferativa de frações semipurificadas e compostos isolados de Croton echioides em células tumorais hepáticas de Rattus norvegicus (HTC) foi avaliada pelo teste MTT. As frações semipurificadas apresentaram citotoxicidade em concentrações acima de 200 µg/mL, em 24, 48 e 72 horas, atingindo viabilidade celular de 24,78% [400 µg/mL] em 24 horas, 12,79% [500 µg/mL] em 48 horas e 10,57% [300 µg/mL] às 72 horas. Para os compostos isolados, o lupeol teve efeito citotóxico em todas as concentrações (1, 5, 10, 15, 20, 40, 60, 80 e 100 µg/mL) e tempos testados (24, 48 e 72 horas), atingindo a viabilidade mínima de 4,37% [100 µg/mL], em 72 horas. Os diterpenos clerodan CEH-1 e CEH-4 também apresentaram atividade antiproliferativa, com viabilidade mínima de 36,19% [100 µg/mL] em 72 horas e 21,33% [100 µg/mL] em 48 horas, respectivamente. No entanto, os diterpenos clerodanos CEH-2 e CEH-3 não apresentaram efeito citotóxico para células HTC. Assim, existe um potencial citotóxico/antiproliferativo de C. echioides contra células tumorais, com enzimas direcionadas a enzimas mitocondriais, associadas à proliferação celular, indicando que esta espécie merece destaque na busca de novas moléculas para o tratamento do câncer.

Antibacterial activity of Barbatimão (Stryphnodendron adstringens) against Staphylococcus aureus: in vitro and in silico studies.

We evaluated the activity of the aqueous fraction and the ethyl acetate fraction of Stryphnodendron adstringens against Staphylococcus aureus and proposed their mechanism of action. The antibacterial activity of S. adstringens fractions was evaluated against S. aureus and the cell targets were rated by docking. The fractions showed moderate antibacterial activity against S. aureus without toxicity on two mammalian cell lines. They also showed synergistic antibacterial activity with tannic acid (TA). In silico assays indicated FabG, FabZ and FabI as probable targets. The metabolic pathway for fatty acid biosynthesis in S. aureus was affected by components of S. adstringens. The synergistic effect when combining TA with S. adstringens fractions suggests a natural alternative to S. aureus control. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study describing the possible targets of action of Stryphnodendron adstringens on Staphylococcus aureus. Molecular dynamics simulations showed that the components of S. adstringens affected the metabolic pathway for fatty acid biosynthesis (FAS II) in S. aureus, inhibiting the FabI, FabG and FabZ enzymes. As tannic acid (TA) is a known inhibitor of some targets identified, we showed synergistic antibacterial activity of S. adstringens in combination with TA. This combination did not show toxicity against HaCaT and Vero cells and based on all these results we suggest that S. adstringens can be a natural and sustainable alternative to S. aureus control.

Desenvolvimento de Maytenus ilicifolia e de seus polifenóis totais sob condição de sombreamento e poda / Shadowing and pruning on the development and polyphenol contents in Maytenus ilicifolia

Maytenus ilicifolia Mart. ex Reissek (Celastraceae), conhecida popularmente por espinheira-santa, é nativa e cultivada, principalmente, no sul do Brasil. Suas folhas apresentam triterpenos e substâncias polifenólicas (flavonóides e taninos) relacionadas ao efeito antiulcerogênico. O objetivo do trabalho foi avaliar a produção de massa seca, altura, ramificação, e o teor de polifenóis totais em plantas de M. ilicifolia que se desenvolveram sob duas condições de luminosidade, após dois tratamentos distintos de poda. O delineamento experimental foi em blocos cazualizados com plantas desenvolvidas sob sombra e a pleno sol, e com plantas manejadas por poda parcial e rasa, com quatro repetições. No tratamento de poda rasa a maior altura das plantas foi observada nas plantas cultivadas a sombra em comparação com as conduzidas a pleno sol. No tratamento de poda parcial o número de ramos terciários aumentou significativamente nas plantas conduzidas a pleno sol. As plantas cultivadas a pleno sol apresentaram diferença significativa na concentração de polifenóis totais em relação às plantas produzidas à sombra alcançando valores de 10,29 ± 0,20% (CV=1,94%) e 7,16 ± 0,09% (CV=1,30%) respectivamente.

The Maytenus ilicifolia Mart. ex Reissek (Celastraceae), traditionally known as "espinheira-santa" in Portuguese, is native and cultivated in South Brazil. Its leaves contain triterpenes and polyphenolic compounds (flavonoids and tannins), which are related to the antiulcerogenic effect. The aim of this work was to evaluate the dried biomass production (g), height (m), ramification, and also the total content in polyphenol compounds of the leaves from the M. ilicifolia specie, which were developed under two different light conditions after two different pruning treatments. The design of the experiment was randomized blocks with two levels of light (shadow or full sunlight) and two levels of prune (partial and drastic), with four replications. The height of the plants grown in the shadow was greater for the treatment of drastic pruning than in plants in full sunlight. The ramification increased significantly with drastic prune in full sunlight. The plants grown in full sunlight showed a significant higher content in total polyphenols than the plants grown in the shadow, 10.29±0.20% (RSD=1.94%) and 7.16±0.09% (RSD=1.30%) respectively.

Validação de metodologia analítica para a determinação de derivados do ácido o-hidroxicinâmico de Echinodorus grandiflorus / Validation of analytical methodology for the determination of derivatives of o-hydroxycinnamic acid from Echinodorus grandiflorus

Este artigo descreve o desenvolvimento e a validação de método espectrofotométrico UV-Visível para quantificação de derivados do ácido o-hidroxicinâmico em folhas de Echinodorus grandiflorus. O método demonstrou ser linear (r² = 0,9974), preciso (DPR < 15%) na análise de matriz complexa e exata (recuperação = 107,56%).

This paper describes the development and the validation of an UV-Vis spectrophotometric method for the quantification of derivatives of o-hydroxycinnamic acid in leaves of Echinodorus grandiflorus. The method showed to be linear (r² = 0.9974), precise (RSD < 15%) in the analysis of complex matrix and accurate (recovery = 107.56%).

Acute and subchronic toxicological evaluation of the semipurified extract of seeds of guaraná (Paullinia cupana) in rodents.

We evaluated the toxicity of a semipurified extract (EPA fraction, containing caffeine and several flavan-3-ols and proanthocyanidins) of seeds of the native Amazon plant Paullinia cupana (guaraná) in rodents. Acute toxicity was tested in male Swiss mice, which received different doses orally (OR) and intraperitoneally (ip); control groups received water. These tests produced acute mortality, with LD(50) of 1.825 g/kg (OR) and 0.827 g/kg (ip), and a significant weight decrease in lungs of mice receiving a dose of 0.1g/kg. In the repeated-dose toxicity test, the EPA was administered OR daily to male and female Wistar rats at doses of 30, 150, and 300 mg/kg/day/90 days. Their behavior, mortality, weight changes, laboratory tests, and the weights and histopathology of organs were evaluated. No rats died during the tests. Males dosed at 150 or 300 mg/kg gained weight more slowly and lost kidney weight (absolute and relative weights, compared to the control group). Hematological and biochemical tests showed few changes, differing somewhat between males and females; the histopathological evaluation indicated no significant changes. These results indicate that the EPA fraction of guaraná caused no toxicity in rats at the smallest dose evaluated (30 mg/kg). No other species was evaluated.

Fitoterápicos: um mercado promissor / Phytomedicines: a promising market

Observa-se na Fitoterapia uma tendência de contribuição efetiva à saúde da população. Por consequência, a padronização de fitomedicamentos é um pré-requisito para a garantia da qualidade, bem como para a constância dos efeitos terapêuticos e segurança do usuário. A validação de processo analítico deve garantir, através de evidências experimentais, que o método atenda às exigências das aplicações analíticas, assegurando a confiabilidade dos resultados. Assim, os equipamentos e materiais de laboratório devem ser devidamente calibrados e o analista qualificado. As substâncias químicas de referência devem ser certificadas por compêndios oficiais, como as Farmacopeias ou por outros códigos autorizados pela legislação vigente. Tão importante quanto o desenvolvimento e validação de uma metodologia analítica é o posterior estudo de estabilidade, a fim de garantir que o produto mantenha sua qualidade durante toda vida útil. Para a obtenção de registro de um medicamento fitoterápico dentro dos padrões requeridos pela legislação faz-se necessário, portanto, a realização de diferentes testes para validação deste medicamento de forma a garantir sua segurança no uso, eficácia na utilização e qualidade do produto.

There is an observable trend towards phytotherapy making a recognized effective contribution to public health. Consequently, the standardization of phytomedicines is a prerequisite for quality assurance and to ensure the consistency of therapeutic effects and safety of the user. Analytical method validation should ensure, through experimental testing, that the method meets the requirements of the analytical applications, ensuring the reliability of results. Thus, the equipment and laboratory materials must be properly calibrated and the analyst qualified. The chemical references must be certified by official compendia, such as pharmacopoeias or other officially accepted codes. As important as the development and validation of an analytical methodology is the stability study, performed to ensure that the product retains its quality throughout its shelf-life. Therefore, to register a herbal medicine in accordance with the legal standards, it is necessary to carry out various validation tests on the product, to ensure its safety, effectiveness and quality.

Effect of lyophilized extracts from guaraná seeds [Paullinia cupana var. sorbilis (Mart.) Ducke] on behavioral profiles in rats.

The aim of the present study was to investigate the pharmacological properties of the crude lyophilized extract (EBPC) of Paullinia cupana seeds (guaraná) and the semi-purified extracts (EPA and EPB) after acute or chronic administration by the oral route in rats. Anxiolytic-like, antidepressant-like and motor stimulant effects were evaluated using the plus maze (PMT), forced swimming (FST) and open field (OFT) tests, respectively. Acute or chronic administration of EBPC (3.0, 30.0 or 60.0 mg/kg) did not alter the percentage of entries or the time spent in the open arm in the PMT. In the FST, chronic treatment with 30.0 mg EBPC/kg and 4.0 mg EPA/kg extract decreased the immobility time similarly to the antidepressant reference drug, imipramine (20.0 mg/kg). Locomotor activity in the OFT was not increased by these extracts. Caffeine (10.0 mg/kg) significantly reduced the immobility time in the FST, but increased locomotor activity in the OFT, indicating psychostimulant activity. The EPB extract did not induce any effect after acute or chronic treatment in the different models used. The present results suggest that the crude EBPC extract and EPA extract produced an antidepressant-like effect after long-term administration.

Ex vivo evaluation of the percutaneous penetration of proanthocyanidin extracts from Guazuma ulmifolia using photoacoustic spectroscopy.

In this work photoacoustic spectroscopy has been applied to determine ex vivo the percutaneous penetration of proanthocyanidins present in extracts obtained from Guazuma ulmifolia, in rats. Lotion formulations containing 0.0663 mg of procyanidin B2 day(-1)animal(-1) were topically applied during 7, 10 and 13 days in each group of the animals. After the end of treatment the animals were killed, the skin dissected to remove the basal content, and the measurements were carried out as a function of the period of time of treatment. The results showed that despite the very low concentration of the active principle (procyanidin B2) in the lotion, the photoacoustic method was able to show the presence of optical absorption bands from this substance in the dermis region, evidencing once again that this method may be useful for studies of topically applied formulations of interest in the pharmacokinetic area.

Estudo mineral das espécies vegetais Alternanthera brasiliana (L.) Kuntze e Bouchea fluminensis (Vell) Mold. / Mineral evaluation of the species Alternanthera brasiliana (L.) Kuntze and Bouchea fluminensis (Vell) Mold.

Alternanthera brasiliana e Bouchea fluminensis são plantas encontradas amplamente na América do Sul, sendo utilizada pela população pelas suas propriedades analgésicas e antiinflamatórias. O trabalho objetivou determinar o teor de cinzas totais e os elementos minerais em ambas espécies. O teor de cinzas totais das folhas de A. brasiliana e B. fluminensis foi de 13,23 por cento ± 0,47 e 8,28 por cento ± 0,07, respectivamente. Em relação à presença de macro e micronutrientes nas folhas observou-se uma maior concentração de nitrogênio (3,13 por cento) e manganês (0,296 por cento) para A. brasiliana e de cálcio (3,08 por cento) e ferro (0,256 por cento) para B. fluminensis. Os dados obtidos, cinzas totais, macro e microelementos, contribuem significativamente no controle de qualidade e padronização de ambas drogas vegetais, além de dar suporte ao uso como suplemento nutricional.

Alternanthera brasiliana and Bouchea fluminensis are plants widely distributed in South America. These plants are used by native populations as analgesic and anti-inflammatory agent. The aim of this paper was to assay the total ashes and mineral content in both species. The levels of total ashes in leaves of A. brasiliana and B. fluminensis were 13.23 percent ± 0.47 and 8.28 percent ± 0.07, respectively. The analysis of macro and micronutrients content in leaves of A. brasiliana shown as main components, nitrogen (3.13 percent) and manganese (0.296 percent). However, in B. fluminensis they were calcium (3.08 percent) and iron (0.256 percent). These data are very useful for the quality control and standardization of the plant raw materials, and support their use as nutritional agents.

Efeitos da intensidade luminosa sobre a morfo-anatomia foliar de Bouchea fluminensis (Vell.) Mold. (Verbenaceae) e sua importância no controle de qualidade da droga vegetal / The effect of light intensity on morpho-anatomy of leaves of Bouchea fluminensis (Vell.) Mold. (Verbenaceae) and their importance on the plant drug control quality

Para a implantação de rigorosos padrões de controle de qualidade das drogas vegetais, além da detecção dos princípios ativos e das metodologias clássicas de microscopia, deve-se também considerar as alterações morfo-anatômicas ocasionadas pelo ambiente, especialmente quando a droga constitui-se de folhas. Neste estudo, foram analisadas as folhas de Bouchea fluminensis cultivadas em pleno sol ou na sombra. As folhas "de sol" são menores do que as "de sombra", com limbo mais ondulado e mais coriáceas ao tato. A análise anatômica revelou espessa cutícula estriada, estômatos anomocíticos em ambas faces, embora raros na face adaxial das folhas de "sombra". Dois tipos de tricomas foram observados, sendo um tector simples com paredes ornamentadas e outro glandular, assim como feixes vasculares colaterais sem tecidos esclerificados, estando os menores envoltos por bainha de células não esclerificadas. A espessura média do mesofilo das folhas "de sol", bem como a altura das células do parênquima paliçádico são significativamente maiores do que nas folhas "de sombra" o que permite diferenciá-las facilmente. A não observância de alterações fotomorfogênicas, nas diversas espécies vegetais utilizadas na forma de droga, pode ocasionar falhas ou dúvidas na identificação das mesmas.

In order to establish rigorous quality control standards for plant drugs, in addition to active substances detection and use of classical microscopical methods, the morpho-anatomical changes caused by the environment must be considered, specially when the plant drug are leaves. In this study, leaves of Bouchea fluminensis cultivated in full sun or in shade were analyzed. Sun leaves are smaller than shade leaves, and their blades are more undulating and leathery to the touch. Anatomical analysis revealed a thick striated cuticle with anomocytic stomates on both faces, although few stomates were present on the adaxial face of shade leaves. Two types of hair were observed, one is a simple tector with ornamented walls, and the other is glandular, as well as collateral vascular sheafs without sclerified tissues, the smaller ones surrounded by a sheath of unsclerified cells. The mean thickness of the mesophyll and the height of the palisade parenchyma cells of sun leaves are significantly higher than in shade leaves, which allows them to be easily distinguished. Failure to observe these photomorphogenic changes in plant species may give misleading results in the identification of plant drugs.

Estudo físico-químico, químico e biológico de extrato das cascas de Stryphnodendron polyphyllum Mart. (Leguminosae)

Two flavan-3-ols and one proanthocyanidins have been isolated from the stem bark of Stryphnodendron polyphyllum Mart., which is traditionally used in Brazil against various diseases. The structure was determined on the basis of spectroscopic data including 1-D (¹H, 13C) and 2-D NMR (¹H/¹H COSY) and MS. The antibacterial activities of an acetone:water and semipurified extracts from the stem bark of Stryphnodendron polyphyllum Mart. were evaluated. Both the crude and semipurified extracts showed activity against Bacillus subtilis and Staphylococcus aureus. Quality control was determined using several pharmacopoeial assay.