Efeitos do D-limoneno no metabolismo energético de camundongos C57/Bl6 com obesidade induzida pela dieta / Effects of D-limonene on the energy metabolism of C57/Bl6 mice with diet-induced obesity

A obesidade está associada ao desenvolvimento de doenças crônicas não transmissíveis como hipertensão, resistência insulínica, dislipidemia e esteatose hepática. O consumo de compostos bioativos impacta na manutenção da saúde e na prevenção de risco de desenvolvimento dessas doenças. Entre os compostos bioativos, os monoterpenos são pouco investigados, apesar da literatura demonstrar efeitos promissores desses compostos sobre o metabolismo. O D-limoneno, o principal monoterpeno encontrado na laranja, é caracterizado por possuir efeitos hipolipemiantes, anti-inflamatórios e anti-obesogênicos. Estudos in vitro e in vivo descrevem sua capacidade de promover a ß-oxidação de ácidos graxos em adipócitos e redução da inflamação. Este estudo teve como objetivo investigar o efeito do D-limoneno no metabolismo e inflamação em um modelo de obesidade induzida por dieta. Para isso, quarenta camundongos machos (C57/Bl6) de 11 semanas de idade, foram distribuídos em 4 grupos, sendo que um dos grupos recebeu ração normolipídica e os demais, ração hiperlipídica. O D-limoneno foi suplementado na ração de dois grupos que receberam dieta hiperlipídica nas concentrações de 0,1%, e 0,8%. Considerando-se a ingestão alimentar dos animais, a ração suplementada com 0,1% D-limoneno correspondeu à ingestão de 0,15 g/kg/dia e ração com 0,8% de D-limoneno correspondeu a 1,3 g/kg/dia. Os animais tiveram o peso e a ingestão alimentar monitorados ao longo da intervenção com duração de 7 semanas. Os camundongos que receberam D-limoneno a 0,1% apresentaram menor ganho de peso e de acúmulo de tecido adiposo, comparado com os animais sem suplementação alimentados com a dieta hiperlipídica. Além disso, o D-limoneno promoveu a diminuição da concentração plasmática de marcadores inflamatórios incluindo TNF-α, INF-γ e IL-6 nos animais dos grupos que foram suplementados com D-limoneno. Entretanto, não houve diferença nos marcadores bioquímicos e metabólicos. Uma limitação do estudo foi o fato das complicações metabólicas associadas ao modelo de obesidade não terem sido plenamente estabelecidas, dados o alojamento individual, à curta duração da exposição à ração hiperlipídica e idade dos animais no início da suplementação. Esse fato pode ter dificultado a observação dos efeitos do D-limoneno na reversão dos parâmetros que seriam normalmente deteriorados pelo desenvolvimento da obesidade. Concluímos que o D-limoneno pode interferir no metabolismo energético, com possível efeito anti-obesogênico e anti-inflamatório. Devido às limitações do modelo, são necessários mais estudos para confirmar esses resultados

Obesity is associated with the development of chronic non-communicable diseases such as hypertension, insulin resistance, dyslipidemia, and hepatic steatosis. The intake of dietary bioactive compounds is associated with the maintenance of health and the prevention of chronic diseases. Among the group of bioactive compounds, monoterpenes are poorly investigated, in spite of several reports of their promising effects on metabolism. D-limonene is the main monoterpene found in oranges, known for its hypolipemic, anti-inflammatory, and anti-obesogenic effects. in vitro and in vivo studies associate D-limonene to increased ß-oxidation of fatty acids in adipocytes and reduced inflammation. This study aimed at investigating the effects of D-limonene on metabolism and inflammation in a diet-induced obesity model. For this purpose, forty male mice (C57/Bl6) were distributed in 4 groups, with one group receiving a normolipidic diet and the others, a high-fat diet. D-limonene was supplemented in the diets of two groups that received high-fat diet at the concentrations of 0.1% and 0.8%. Considering the feed intake, mice receiving D-limonene supplementation at 0.1% ingested in average 0.15 g/kg/day, while the mice receiving the supplemmentation at 0.8%, ingested approximately 1.3 g of D-limonene /kg/day. The animals had their weight and food intake monitored throughout the intervention. Mice that received Dlimonene supplementation at 0.1% showed reduced weight gain and accumulation of adipose tissue compared to the non-supplemented mice fed the high-fat diet. In addition, D-limonene promoted a decrease in hepatic inflammatory markers including TNF-α, INF-γ, and IL-6. However, there was no difference in biochemical and metabolic markers. A limitation of the study was that the metabolic complications associated with the obesity model were not fully established, probably due to the age at the start of the protocol (11 weeks), individual housing and short duration of the exposure to the high-fat feed. This fact may have prevented the observation of the positive effects of D-limonene in reversing parameters that would normally be impaired by the development of obesity. We conclude that D-limonene may interfere in energy metabolism, with a possible anti-obesogenic and anti-inflammatory effect. Due to the limitations of the model, further studies are needed to confirm these findings

A (-)-Carvona Modula a Sinalização de Cálcio Intracelular com Ação Antiarrítmica em Corações de Ratos / (-)-Carvone Modulates Intracellular Calcium Signaling with Antiarrhythmic Action in Rat Hearts

Resumo Fundamento: A (-)-carvona é um monoterpeno encontrado em óleos essenciais com atividade antioxidante e anti-inflamátoria. Objetivos: O objetivo deste estudo foi analisar a propriedade antiarrítmica da (-)-carvona no coração de rato e seus efeitos sobre a sinalização de Ca+2 intracelular. Métodos: Os efeitos da (-)-carvona foram avaliados sobre a contratilidade atrial (0,01 - 4 mM) e ventricular (0,5 mM), e no eletrocardiograma (0,5mM). A fração de encurtamento, a corrente de cálcio do tipo L (ICa,L) e a sinalização de Ca+2 foram medidas no cardiomiócito isolado (0,5 mM). O efeito antiarrítmico foi avaliado no modelo de arritmia induzida por sobrecarga de cálcio (0,5 mM) (n = 5). Um p < 0,05 foi adotado como nível de significância estatística. Resultados: No átrio, a (-)-carvona causou inotropismo negativo de maneira concentração-dependente (EC50 0,44 ± 0,11 mM) e diminuiu o inotropismo positivo induzido pelo CaCl2 (0,1 - 8,0 mM) e BAY K8644 (5 - 500 nM), um agonista de canal de cálcio do tipo L. Em coração isolado, a (-)-carvona (0,5mM) reduziu a contratilidade ventricular em 73% e a frequência cardíaca (em 46%), aumentou o Pri (30,7%, tempo desde o início da onda P até a onda R) e o QTc (9,2%, uma medida de despolarização e repolarização dos ventrículos), sem mudar a duração do complexo QRS. A (-)-carvona diminuiu a fração de encurtamento (61%), a (ICa,L) (79%) e o transiente intracelular de Ca+2 (38%). Além disso, a (-)-carvona apresentou ação antiarrítmica, identificada pela redução do escore de arritmia (85%) e ocorrência de fibrilação ventricular. Conclusão: A (-)-carvona reduz a entrada de Ca+2 através de canais de Ca+2 do tipo L e, assim, diminui a contratilidade cardíaca e o Ca+2 intracelular e apresenta promissora atividade antiarrítmica no coração de ratos.

Abstract Background: (-)-Carvone is a monoterpene found in essential oils with antioxidant and anti-inflammatory activity. Objective: The aim of this paper was to analyze the antiarrhythmic property of (-)-carvone in the rat heart and its effects on the intracellular Ca2+ signaling. Methods: The effects of (-)-carvone were evaluated on the ventricular (0.5 mM) and atrial contractility (0.01 - 4 mM) and on electrocardiogram (0.5 mM). Fractional shortening, L-type calcium current (ICa,L) and Ca2+ signaling were measured in the isolated cardiomyocyte (0.5 mM). Antiarrhythmic effect was evaluated in arrhythmia model induced by calcium overload (0.5 mM) (n = 5). P < 0.05 was used as the significance level. Results: In the atrium, (-)-carvone evoked negative inotropism that was concentration-dependent (EC50 0.44 ± 0.11 mM) and decreased the positive inotropism evoked by CaCl2 (0.1 to 8.0 mM) or BAY K8644 (5 to 500 nM), an agonist of L-type Ca2+ channel. In isolated heart, (-)-carvone (0.5 mM) promoted reduction of ventricular contractility (73%) and heart rate (46%), increased PRi (30.7%, time from the onset of the P wave until the R wave) and QTc (9.2%, a measure of the depolarization and repolarization of the ventricles) without changing the QRS complex duration. (-)-Carvone decreased the fractional shortening (61%), ICa,L (79%) and Ca2+ intracellular transient (38%). Furthermore, (-)-carvone showed antiarrhythmic action, verified by decrease of the arrhythmia score (85%) and occurrence of ventricular fibrillation. Conclusion: (-)-Carvone decreases Ca2+ entry through L-type Ca2+ channels, reducing the cardiac contractility and intracellular Ca2+, and, therefore, presenting promising antiarrhythmic activity in the rat hearts.

Avaliação in vitro e in silico da atividade do S-(-)-Álcool Perílico sobre linhagens celulares do Carcinoma Epidermóide de Língua / In vitro and in silico evaluation of the activity of S-(-)-Peryllic Alcohol on Tongue Squamous Carcinoma cell lines

O carcinoma epidermóide oral (CEO) é a neoplasia maligna mais frequente da cavidade oral e constitui um problema de saúde pública devido a sua alta taxa de incidência e mortalidade devido em muitos casos ao fracasso terapêutico e a resistência tumoral. Assim sendo, destaca-se a busca por novas moléculas biologicamente ativas, como as encontradas nos produtos de origem natural. Este trabalho tem como objetivo avaliar a atividade antineoplásica do S-(-)-álcool perílico (POH) em culturas de células de CEO de língua e predizer sua afinidade através de modelo computacional sobre proteínas que regulam o ciclo celular. Para isso, foram utilizadas duas linhagens celulares de CEO de língua, HSC-3 e SCC-25. Os seguintes grupos foram analisados: G0 (controle; células cultivadas na ausência de POH), G1 (células tratadas com cisplatina a 40 µM), G2 (células tratadas com POH a 0,5 mM), G3 (células tratadas com POH a 1,0 mM), G4 (células tratadas com POH a 1,5 mM) e G5 (células tratadas com POH a 3,0 mM). Diferenças entre estes grupos foram investigadas através dos seguintes ensaios: viabilidade celular (Alamar Blue e Live/Dead assay) e atividade migratória (Wound healing). Foi também realizada a predição de afinidade entre o POH e as moléculas de controle do ciclo celular utilizando a docagem molecular com emprego do software Molegro Virtual Docker, v. 6.0.1. Os dados foram tratados estatisticamente pelo GraphPad Prism 6.0 (GraphPad Software, EUA), análises paramétricas utilizando teste Anova, pós-teste de Tukey e teste estatístico não-paramétricos de Kruskal-Wallis, seguido pelo teste t de estudent foram adotados para determinação de diferenças entre os grupos experimentais. O índice de significância considerado neste trabalho foi de 5%. Para ambas as técnicas de avaliação da viabilidade celular (Alamar Blue e Live/dead assay) analisadas neste trabalho, o POH foi capaz de reduzir a viabilidade celular de linhagens do CEO de língua de maneira dosedependente e tempo-dependente (p<0,05). As concentrações de 1,5 mM e 3 mM do POH obtiveram resultados melhores ou semelhantes aos encontrados na cisplatina 40 µM, para as duas linhagens, na avaliação da viabilidade celular (p<0,05). Os valores de IC50 do POH foram de 1,5 mM para a célula SCC-25 em todos os intervalos de tempo (24 h, 48 h e 72 h), uma vez que, para a linhagem HSC-3, foram de 3 mM para os tempos de 24 h e 48 h e de 1,5 mM para o intervalo de 72 h. O POH foi capaz de inibir a migração das duas linhagens celulares de CEO de maneira dependente da concentração (p≤0,05), comparados ao grupo controle. A habilidade da molécula POH se ligar a proteínas responsáveis pela ativação do ciclo celular foi avaliada usando docking models. Dentre elas, a proteína GTPase Kras mostrou a melhor energia de ligação (-86.70 kcal/mol), apresentando ligações de hidrogênio com os resíduos THR58 (A) e ASP57 (A) e ligações estéricas com os resíduos TRY32 (A) e ALA18 (A). As evidências deste estudo corroboram a ideia de que o POH possui atividade sobre o CEO, sugerindo que essa molécula possa ser uma forte candidata para o desenvolvimento de medicamentos direcionados ao tratamento desta patologia (AU).

Oral squamous cell carcinoma (OSCC) is the most frequent malignant neoplasm of the oral cavity and constitutes a public health problem due to its high incidence and mortality rate caused in many cases by therapeutic failure and tumor resistance. Therefore, the search for new biologically active molecules stands out, such as those found in products of natural origin. This work aims to evaluate the antineoplastic activity of S-(-)-perillyl alcohol (POH) in cell cultures of tongue CEO and to predict its affinity through a computer model on proteins that regulate the cell cycle. For this purpose, two cell lines of tongue CEO were used, HSC-3 and SCC-25. The following groups were analyzed: G0 (control; cells cultured in the absence of POH), G1 (cells treated with 40 µM cisplatin), G2 (cells treated with 0.5 mM POH), G3 (cells treated with 1 .0 mM), G4 (cells treated with 1.5 mM POH) and G5 (cells treated with 3.0 mM POH). Differences between these groups were investigated through the following assays: cell viability (Alamar Blue and Live/Dead assay) and migratory activity (Wound healing). Affinity prediction between POH and cell cycle control molecules were also performed using molecular docking using Molegro Virtual Docker, v. 6.0.1. The data was statistically treated by GraphPad Prism 6.0 (GraphPad Software, USA), parametric analysis using Anova test, Tukey post-test and Kruskal-Wallis non-parametric statistical test, followed by t student test were adopted for determination of differences between the experimental groups. The significance index considered in this work was 5%. For both cell viability assessment techniques (Alamar Blue and Live/dead assay) analyzed in this work, POH was able to reduce the cell viability of tongue CEO lines in a dose-dependent and time-dependent manner (p<0 .05). The concentrations of 1.5 mM and 3 mM of POH obtained better or similar results to those found in 40 µM cisplatin, for the two strains, in the evaluation of cell viability (p<0.05). The IC50 values of POH were 1.5 mM for the SCC-25 cell at all time intervals (24 h, 48 h and 72 h), since for the HSC-3 line they were 3 mM for 24 h and 48 h times and 1.5 mM for the 72 h interval. POH was able to inhibit the migration of the two DSC cell lines in a concentration-dependent manner (p≤0.05), compared to the control group. The ability of the POH molecule to bind to proteins responsible for cell cycle activation was evaluated using docking models. Among them, the protein GTPase Kras showed the best binding energy (-86.70 kcal/mol), featuring hydrogen bonds with residues THR58 (A) and ASP57 (A) and steric bonds with residues TRY32 (A) and ALA18 ( THE). The evidence from this study supports the idea that POH has antineoplastic activity on the CEO, suggesting that this molecule may be a strong candidate for the development of drugs aimed at the treatment of this pathology (AU).

Chemical composition and larvicidal activity of the essential oil of Pimenta dioica leaves / Composición química y actividad larvicida del aceite esencial de hojas de Pimenta dioica

In this study, we investigated the main constituent, the predominant class and biological activity of the essential oil extracted from the leaves of Pimenta dioica and the pattern of the major constituent against larvae in the third stage of Aedes aegypti. For this reason, we extracted the oil by hydrodistillation, identified its components by gas chromatography coupled with mass spectrometry (GC/MS) and calculated the lethal concentration (LC50) of the larvicidal activity using the Reed-Muench method. The results show that the oil consists mainly of eugenol, in which the phenylpropanoid class predominated and the lethal concentration, LC50, was 38.86 µg mL-1at a confidence level of 2.25 µg mL-1, while the eugenol standard presented LC5079.75 µg mL-1at a confidence level of 2.10 µg mL-1. Given the facts, we conclude that the oil is more active than the standard and that it has the potential to replace chemical larvicides.

En este estudio, investigamos el constituyente principal, la clase predominante y la actividad biológica del aceite esencial extraído de las hojas de Pimenta dioica y el patrón del constituyente principal contra las larvas en la tercera etapa de Aedes aegypti. Por este motivo, extrajimos el aceite por hidrodestilación, identificamos sus componentes mediante cromatografía de gases acoplada a espectrometría de masas (GC/MS) y calculamos la concentración letal (CL50) de la actividad larvicida mediante el método Reed-Muench. Los resultados muestran que el aceite está constituido principalmente por eugenol, en el que predominó la clase fenilpropanoide y la concentración letal, CL50, fue de 38,86 µg.mL-1 a un nivel de confianza de 2,25 µg.mL-1, mientras que el estándar de eugenol presentó CL50 79,75 µg.mL -1 a un nivel de confianza de 2,10 µg.mL-1. Dados los hechos, concluimos que el aceite es más activo que el estándar y que tiene el potencial de reemplazar los larvicidas químicos.

Dynamic control of ERG20 expression to improve production of monoterpenes by engineering Saccharomyces cerevisiae / 中国中药杂志

Monoterpenes are widely used in cosmetics, food, medicine, agriculture and other fields. With the development of synthetic biology, it is considered as a potential way to create microbial cell factories to produce monoterpenes. Engineering Saccharomyces cerevisiae to produce monoterpenes has been a research hotspot in synthetic biology. In S. cerevisiae, the production of geranyl pyrophosphate(GPP) and farnesyl pyrophosphate(FPP) is catalyzed by a bifunctional enzyme farnesyl pyrophosphate synthetase(encoded by ERG20 gene) which is inclined to synthesize FPP essential for yeast growth. Therefore, reasonable control of FPP synthesis is the basis for efficient monoterpene synthesis in yeast cell factories. In order to achieve dynamic control from GPP to FPP biosynthesis in S. cerevisiae, we obtained a novel chassis strain HP001-pERG1-ERG20 by replacing the ERG20 promoter of the chassis strain HP001 with the promoter of cyclosqualene cyclase(ERG1) gene. Further, we reconstructed the metabolic pathway by using GPP and neryl diphosphate(NPP), cis-GPP as substrates in HP001-pERG1-ERG20. The yield of GPP-derived linalool increased by 42.5% to 7.6 mg·L~(-1), and that of NPP-derived nerol increased by 1 436.4% to 8.3 mg·L~(-1). This study provides a basis for the production of monoterpenes by microbial fermentation.

Metabolitos bioactivos y actividad antioxidante in vitro del aceite esencial extraído de dos especies del género Tagetes

Objetivo: este estudio investigó los metabolitos bioactivos y actividad antioxidante de aceites esenciales de dos especies del género Tagetes. Método: el aceite esencial se obtuvo por arrastre a vapor de agua, posteriormente se determinó su rendimiento de extracción, densidad relativa, índice de refracción y su solubilidad en etanol (70 % v/v). La composición química fue evaluada mediante cromatografía de gases acoplada a espectrometría de masas (GC-MS). La actividad antioxidante fue determinada mediante el método del radical libre 2,2-difenil-1-picrilhidracilo (DPPH) y la capacidad de atrapamiento del catión radical ABTS*+. Resultados: en los aceites esenciales de las especies Tagetes se logró identificar 26 componentes químicos para la especie Tagetes elliptica Sm. y 16 para Tagetes minuta L., ambas especies presentaron como componentes principales a los monoterpenos (61 %) y sesquiterpenos (44 %). Los metabolitos bioactivos de aceites esenciales entre ambas especies de Tagetes fueron, (3-trans-ocimeno (25,03 %), transtagetona (51,37 %), ß-mirceno (2,78) y ß-cariofileno (1,17 %). Los rendimientos de extracción oscilaron entre 0,05 y 0,048 %, la densidad entre 0,90 y 0,88 (g/ml) con un índice de refracción de 1,493 y 1,482 y una solubilidad (v/v) positiva entre ambas especies. La actividad antioxidante del aceite esencial en ambas especies mostró una variación entre 1,77 y 2,56 mg/mL para el DPPH y 21,02 a 41,06 mg/mL para BTS*+. Conclusión: los aceites esenciales de las especies Tagetes elliptica Sm. y Tagetes minuta L. son una fuente de metabolitos bioactivos fomentan potencialidades antimicrobianas y antioxidantes con fines de su uso como conservantes alimentarios.

SUMMARY Aim: This study investigated the bioactive metabolites and antioxidant activity of essential oils of two species of the genus Tagetes. Method: The essential oil was obtained by steam stripping, its extraction performance, relative density, refractive index and its solubility in ethanol (70 % v/v) were subsequently determined. The chemical composition was tested using gas chromatography coupled to mass spectrometry (GC-MS). The antioxidant activity was determined using the 2,2-diphenyl-1-picrylhydracil (DPPH) free radical method and the ABTS*+ radical cation trapping capacity. Results: In the essential oils of the Tagetes species, it was found to identify 26 chemical components for the species Tagetes elliptica Sm. And 16 for Tagetes minuta L., both species presented monoterpenes (61 %) and sesquiterpenes (44 %) as main components. The bioactive metabolites of essential oils between both species of Tagetes were, (-trans-ocimeno (25.03 %), trans-tagetone (51.37 %), ß-myrcene (2.78) and ß-caryophyllene (1.17 %). The extraction yields ranged between 0.05 and 0.048 %, the density between 0.90 and 0.88 (g/ml) with a refractive index of 1.493 and 1.482 and a positive solubility (v/v) between both species. The antioxidant activity of the essential oil in both species showed a variation between 1.77 and 2.56 mg/mL for DPPH and 21.02 to 41.06 mg/mL for BTS*+. Conclusion: The essential oils of the species Tagetes elliptica Sm. and Tagetes minuta L. are a source of bioactive metabolites that promote antimicrobial and anti-oxidant potentialities for use as food preservatives.

Objetivo: este estudo investigou os metabólitos bioativos e a atividade antioxidante de óleos essenciais de duas espécies do gênero Tagetes. Método: o óleo essencial foi obtido por decapagem a vapor, posteriormente foram determinados seu desempenho de extração, densidade relativa, índice de refração e sua solubilidade em etanol (70% v/v). A composição química foi trabalhada por cromatografía gasosa acoplada a espectrometría de massas (GC-MS). A atividade antioxidante foi determinada pelo método do radical livre 2,2-difenil-1-picrilidracila (DPPH) e pela capacidade de captura do cátion radical ABTS* +. Resultados: nos óleos essenciais das espécies Tagetes, identificou-se 26 componentes químicos para a espécie Tagetes elliptica Sm. E 16 para Tagetes minuta L., ambas as espécies apresentaram monoterpenos (61%) e sesquiterpenos (44%) como principais componentes. Os metabólitos bioativos dos óleos essenciais entre as duas espécies de Tagetes foram, (3-trans-ocimeno (25,03%), transtagetona (51,37%), ß-mirceno (2,78) e ß-cariofileno (1,17%). Os rendimentos de extração variaram entre 0,05 e 0,048%, a densidade entre 0,90 e 0,88 (g/ml) com índice de refração de 1,493 e 1,482 e solubilidade positiva (v/v) entre as duas espécies. A atividade antioxidante do óleo essencial em ambas as espécies apresentou variação entre 1,77 e 2,56 mg/mL para DPPH e 21,02 a 41,06 mg/mL para BTS* +. Conclusão: os óleos essenciais das espécies Tagetes elliptica Sm. e Tagetes minuta L. são uma fonte de metabólitos bioativos, atualmente com potencial antimicrobiano e antioxidante com a finalidade de serem conservantes de alimentos.

O potencial antimicrobiano do carveol: uma revisão integrativa

Objetivo: realizar uma revisão bibliográfica sobre o potencial antimicrobiano do carveol. É uma revisão integrativa entre os anos de 2010-2020 nas bases de dados BVS, PubMed, SciELO, ScienceDirect e Web of Science. Metodologia: dos 2753 artigos recuperados nas bases de dados, apenas 5 foram selecionados para compor os resultados, sendo os gêneros mais estudados Candida, Staphylococcus e Escherichia. A técnica de microdiluição em caldo foi a metodologia mais utilizada para determinar sua ação. Em termos de MIC, para a maioria das espécies, foram excelentes. Em relação ao teste Time-kill, o carveol teve um excelente desempenho na concentração de 4 x MIC, que mostrou uma redução de UFC de 6 log em 2 horas. Não foram encontrados na literatura estudos sobre a utilização do método Checkboard. Além disso, este composto apresentou boa atividade antibiofilme contra bactérias Staphylococcus aureus. Resultados: o possível mecanismo de ação da atividade antimicrobiana do carveol ocorre por meio dos graves danos causados às membranas citoplasmáticas, o que favorecerá a lise celular. Dessa forma, existem poucos estudos na coleção científica disponível que abordem o assunto, sendo necessária a realização de mais estudos in vitro e in vivo para melhor elucidar sua ação.

SUMMARY Aim: to carry out a bibliographic review on the antimicrobial potential of carveol. It is an integrative review between the years 2010-2020 in the BVS, PubMed, SciELO, Science-Direct and Web of Science databases. Methodology: of the 2753 articles retrieved from the databases, only 5 were selected to compose the results, the most studied genera being Candida, Staphylococcus and Escherichia. The broth microdilution technique was the most widely used methodology to determine its action. In terms of MIC, for most species, they were excellent. Regarding the Time-kill test, carveol had an excellent performance at a concentration of 4 x MIC, which showed a reduction of CFU by 6 log in 2 hours. No studies have been found in the literature on the use of the Checkboard method. Further-more, this compound had good antibiofilm activity against Staphylococcus aureus bacteria. Results: the possible mechanism of action of the antimicrobial activity of carveol occurs through the severe damage caused to the cytoplasmic membranes, which will favor cell lysis. Thus, there are few studies in the available scientific collection that address the subject, so it is necessary to carry out more in vitro and in vivo studies to better elucidate its action.

Objetivo: realizar una revisión bibliográfica sobre el potencial antimicrobiano del carveol. Se trata de una revisión integradora entre los años 2010-2020 en las bases de datos de BVS, PubMed, SciELO, ScienceDirect y Web of Science. Metodología: de los 2753 artículos recuperados de las bases de datos, solo se seleccionaron 5 para componer los resultados, siendo los géneros más estudiados Candida, Staphylococcus y Escherichia. La técnica de microdilución en caldo fue la metodología más utilizada para determinar su acción. En cuanto a CIM, para la mayoría de las especies, fueron excelentes. En cuanto al ensayo Time-kill, el carveol tuvo un excelente desempeño a una concentración de 4 x MIC, que mostró una reducción de UFC en 6 log en 2 horas. No se ha encontrado ningún estudio en la literatura sobre el uso del método Checkboard. Además, este compuesto tenía una buena actividad antibiofilm contra la bacteria Staphylococcus aureus. Resultados: el posible mecanismo de acción de la actividad antimicrobiana del carveol se produce a través del severo daño causado en las membranas citoplasmáticas, lo que favorecerá la lisis celular. Así, son pocos los estudios en la colección científica disponible que abordan el tema, por lo que es necesario realizar más estudios in vitro e in vivo para dilucidar mejor su acción.

Antimicrobial activity of the essential oils from the leaves and stems of Amomum rubidum Lamxay & N. S. Lý / Actividad antimicrobiana de los aceites esenciales de las hojas y tallos de Amomum rubidum Lamxay & N. S. Lý

This paper described the chemical compositions and antimicrobial activity of the essential oils from the leaves and stem of Amomum rubidumLamxay & N. S. Lý, collected from Bidoup Nui Ba National Park, Lam Dong, Vietnam. The essential oils were obtained by hydrodisitllation method while antimicrobial activity was evaluetd by microdilution broth susceptibility assay. The main constituents of the leaf essential oil were identified as 1,8-cineole (37.7%), δ-3-carene (19.5%) and limonene (16.3%) while δ-3-carene (21.9%), limonene (17.8%) and ß-phellandrene (14.6%) dominated in the stem essentialoil. The leaf and stem essential oils displayed stronger inhibition of Pseudomonas aeruginosa with MIC of 25 µg/mLand 50 µg/mL respectively. The stem essential oil was active against Candida albicans (MIC, 50 µg/mL) while both essential oils inhibited the growth of Fusarium oxysporum (MIC 50 µg/mL). This is the first report on chemical constituents and antimicrobial activity of the essential oils of A. rubidum.

Este artículo describe la composición química y la actividad antimicrobiana de aceites esenciales de las hojas y el tallo de Amomum rubidum Lamxay & N. S. Lý recolectados del Parque Nacional Bidoup Nui Ba, Lam Dong, Vietnam. Los aceites esenciales se obtuvieron mediante el método de hidrodisitilación, mientras que la actividad antimicrobiana se evaluó mediante un ensayo de susceptibilidad de caldo de microdilución. Los principales componentes del aceite esencial de la hoja se identificaron como 1,8-cineol (37,7%), δ-3-careno (19,5%) y limoneno (16,3%), mientras que δ-3-careno (21,9%), limoneno (17,8 %) y ß-felandreno (14,6%) dominaron en el aceite esencial del tallo. Los aceites esenciales de hoja y tallo mostraron una inhibición más fuerte de Pseudomonas aeruginosa con un MIC de 25 µg/mL y 50 µg/mL, respectivamente. El aceite esencial del tallo fue activo contra Candida albicans (MIC, 50 µg/mL) mientras que ambos aceites esenciales inhibieron el crecimiento de Fusarium oxysporum (MIC 50 µg/mL). Este es el primer informe sobre los componentes químicos y la actividad antimicrobiana de los aceites esenciales de A. rubidum.

Extracción por microondas libre de solvente del aceite esencial de naranja (Citrus sinensis), y el efecto de las condiciones de proceso en su rendimiento, composición y actividad antimicrobiana

RESUMEN La extracción asistida por microondas (MAE por sus siglas en inglés) es una técnica eco amigable relativamente nueva que ha recibido una atención creciente debido a su menor tiempo de extracción y consumo de energía, mayor rendimiento de compuestos de interés y menor consumo de disolvente; a diferencia de técnicas convencionales de extracción, que implican elevados gastos de energía y tiempo. Así mismo, subproductos de industrias como cáscaras de cítricos, son desechados en grandes cantidades, no aprovechándose las propiedades funcionales sus compuestos. En esta investigación se realizó la extracción de aceite esencial a partir de cáscara de naranja, por microondas libre de solventes (SFME por sus siglas en inglés), a diferentes condiciones de potencia (400, 800 W, y sus combinaciones). El objetivo fue evaluar el efecto de las diferentes condiciones de proceso sobre el rendimiento del aceite esencial extraído, sobre su composición química y actividad antimicrobiana. Los más altos rendimientos de extracción correspondieron a los aceites extraídos a 400 W y a 800/400 W, siendo este último el que conllevó un menor gasto de energía y emisión de CO2. En general, todos los aceites extraídos presentaron los mismos compuestos mayoritarios (limoneno, β- mirceno, linalool y α- pineno), mientras que el aceite sometido a 400 W, presentó cantidades menores de 0,23% de 5 compuestos minoritarios adicionales. Sobre la actividad antimicrobiana, se detectó mayor actividad contra Listeria monocytogenes (ATCC 19115) respecto a Escherichia coli (ATCC 8739) en todos los tratamientos.

ABSTRACT Microwave assisted extraction (MAE) is a relatively new ecofriendly technique that has received increasing attention due to its reduced extraction time and energy consumption, higher yield and lower solvent consumption; unlike conventional extraction techniques, which involve higher energy costs and greater time. Likewise, industry by-products such as citrus peels are disposed of in large quantities, thus functional properties of their compounds are left unused. In the present investigation, solvent-free microwave extraction (SFME) was used to obtain orange peel essential oil at different power conditions (400, 800 W, and their combinations). The aim was to evaluate the effect of the process on the yield, composition and antimicrobial activity of the essential oil obtained. The highest extraction yields were obtained using powers of 400 and 800/400 W, corresponding the minor energy and CO2 emission generated to the latter. In general, all oils had the same major components (limonene, β- myrcene, linalool and α- pinene), with the oil obtained at 400 W having, in addition, five minor compounds, in amounts less than 0.23%. Regarding antimicrobial activity, in general, greater activity against L. monocytogenes (ATCC 19115) was detected in comparison with E. coli (ATCC 8739) in all treatments.

Synergistic antibacterial activity of monoterpenes in combination with conventional antimicrobials against Gram-positive and Gram-negative bacteria / Atividade antibacteriana sinérgica de monoterpenos combinados com antimicrobianos convencionais frente a bactérias Gram-positivas e Gram-negativas

Introduction: bacterial infections are a public health problem. Besides, the emergence of strains resistant to antimicrobials has contributed to the search for new alternatives, such for the terpenes with antimicrobial potential. Objectives: the objective of this study was to determine the possible interaction of isolated monoterpenes (-)-Carveol, Geraniol, Citronellol, α-terpineol, R-(-) Carvone, (-)-Menthol, Linalool, D-Dihydrocarvone, and (-)-Terpine-4-ol with conventional antimicrobials (Chloramphenicol, Minocycline, Amoxicillin and Ciprofloxacin) when they are evaluated on Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa strains. Methodology: the minimum inhibitory concentrations of these test drugs were determined using the microdilution method. The Checkerboard method was used to assess the interactions, by determining the fractional inhibitory concentration index (FIC index). Results: aamong the monoterpenes, only Carveol, Citronellol, and Geraniol presented antimicrobial activity (MIC < 1024 µg/mL). They presented synergistic effects against Pseudomonas aeruginosa ATCC-9027 (FIC index ≤ 0.5) when in combination with Minocycline. Conclusion: this study contributes to the development of new approaches to control bacterial resistance and to the possibility of discovering new drugs.

Introdução: as infecções bacterianas são um problema de saúde pública. Além disso, o surgimento de cepas resistentes aos antimicrobianos tem contribuído para a busca de novas alternativas, como a pesquisa de terpenos com potencial antimicrobiano. Objetivos: o objetivo deste estudo foi determinar a possível interação de monoterpenos isolados (-) - Carveol, Geraniol, Citronelol, α-terpineol, R - (-) Carvona, (-)-Mentol, Linalol, D-Diidrocarvona e (-)-Terpina-4-ol com antimicrobianos convencionais (cloranfenicol, minociclina, amoxicilina e ciprofloxacina) quando avaliados em Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, Escherichia coli e Pseudomonas aeruginosa. Metodologia: as concentrações inibitórias mínimas destas drogas foram determinadas usando o método de microdiluição. O método checkerboard foi utilizado para avaliar as interações, determinando o índice de concentração inibitória fracionária (índice FIC). Resultados: entre os monoterpenos, apenas Carveol, Citronelol e Geraniol apresentaram atividade antimicrobiana (CIM < 1024 µg/mL). Eles apresentaram efeitos sinérgicos contra Pseudomonas aeruginosa ATCC-9027 (índice FIC ≤ 0,5) quando em combinação com Minociclina. Conclusão: este estudo contribui para o desenvolvimento de novas abordagens para o controle da resistência bacteriana e para a possibilidade de descoberta de novas drogas.

Toxicity of the essential oil of basil cultivars and hybrids and its repellent effect on stored grain pests / Toxicidade do óleo essencial de cultivares e híbridos de manjericão e seu efeito repelente sobre pragas de grãos armazenados

Essential oils have emerged as an alternative to synthetic insecticides in the control of stored grain pests. The toxicity and repellency of the essential oils of four basil cultivars and three basil hybrids and the monoterpenes linalool, citral, and (E)-methyl cinnamate were evaluated in the stored grain pests Callosobruchus maculatus and Sitophillus zeamais. The essential oils of the cultivar Genovese and the hybrid 'Genovese' x 'Maria Bonita' were more toxic to C. maculatus. Conversely, the essential oils of the cultivar Sweet Dani and the hybrid 'Cinnamom' x 'Maria Bonita' were more toxic to S. zeamais. Among the monoterpenes, (E)-methyl cinnamate was the most toxic to both pests, taking 0.14 and 0.34 µ L.mL-1 to kill 50% of the C. maculatus and S. zeamais populations, respectively. All essential oils from cultivars, hybrids, and monoterpenes were repellent to S. zeamais, except for (E)-methyl cinnamate. For C. maculatus, this effect was lower, being citral the most repellent compound. Results demonstrate the insecticidal potential of the essential oil of O. basilicum and its monoterpenes in the control of stored grain pests.

Os óleos essenciais surgem como alternativa aos inseticidas sintéticos no controle das pragas de grãos armazenados. A toxicidade e a repelência dos óleos essenciais de quatro cultivares e três híbridos de manjericão e dos monoterpenos linalol, citral e (E)-cinamato de metila foram avaliadas nas pragas de grãos armazenados Callosobruchus maculatus e Sitophillus zeamais. Os óleos essenciais da cultivar Genovese e do híbrido 'Genovese' x 'Maria Bonita' foram mais tóxicos para C. maculatus. Já para S. zeamais, os óleos essenciais das cultivar Sweet Dani e do híbrido 'Cinnamom' x 'Maria Bonita' apresentaram maior toxicidade. Dentre os monoterpenos, o (E)-cinamato de metila foi o mais tóxico para ambas as pragas. Foram necessários 0,14 e 0,34 µ L.mL-1 para matar 50% da população de C. maculatus e S. zeamais. Todos os óleos essenciais das cultivares, dos híbridos e dos monoterpenos foram repelentes a S. zeamais, com exceção do (E)-cinamato de metila. Já para C. maculatus, este efeito foi reduzido, sendo o citral o composto mais repelente. Nos resultados demonstram o potencial inseticida dos óleos essenciais de O. basilicum e seus monoterpenos para o controle de pragas de grãos armazenados.

Elettaria Cardamom Extract Synergistically Enhaning Sorafenib-Induced Apoptosis in HepG2 Cells

Introduction: Liver cancer is ranked as the second most common cause of death globally as a result of its poorprognosis. It can be treated with sorafenib, but its use is limited due to its toxicity and adverse reactions. Lowerdoses of sorafenib with other complementary agents are recommended to minimize toxicity. Cardamom seeds areone of the most common ingredients of Indian and Chinese traditional medicine, and different studies havesuggested that cardamom extract can display anti-cancer activities. Aim: this study aims to investigate theefficiency of Elettaria Cardamom Extract (ECE) on enhancement of Sorafenib-induced apoptosis in HepG2.Methods: Human liver cancer cell line (HepG2) were exposed to increasing concentrations of individual andcombined treatments of Sorafenib and ECE for 24 h. The viability of cells was examined using MTT Assay.Clonogenicity and cell migration assays were carried out. Reactive oxygen species (ROS) generation andmitochondrial membrane potential (MMP) level were determined by DCFH-DA and JC-1 dye, respectively.Agarose gel electrophoresis and comet examinations were carried out to estimate the DNA damage. Results:Combined treatment of ECE with sorafenib suppressed the proliferation, colony formation and cell migration ofHepG2 cells more than the sorafenib did alone. The half maximal inhibitory concentration (IC50), after 24h ofincubation were 15 µM of sorafenib and 9 and 7.3 µM of sorafenib enhanced by 5 and 10 µg / 100 µl of ECErespectively. HepG2 treated cells displayed biochemical features of apoptotic cell death. The combined treatmentincreased the ROS production, reduced the level of MMP, increased Comet tail length and induced DNAfragmentation more than sorafenib did alone. Conclusions: These findings demonstrate that ECE enhanced thesorafenib effect in HepG2 cells and suggest that the ECE may be a promising agent for reducing sorafenib sideeffects in hepatocellular carcinoma (HCC).

Research progress on processing history evolution, chemical constituents and pharmacological action of Paeoniae Radix Alba / 中草药

Paeoniae Radix Alba (PRA) is a clinically commonly used drug for tonifying blood and regulating meridians, which was listed as one of the highest grade herb in Shengnong's Classic of Materia Medica. The main chemical constituents of PRA are monoterpenes, triterpenes and flavonoids, and its processing methods are cleansing, cutting, adding auxiliary materials, etc. At present, the mainstream processing methods are stir-frying or stir-frying with wine. Modern studies have shown that PRA has the effects of protecting cardiovascular, anti-inflammatory and anti-oxidant. In this paper, we consulted with the ancient Chinese materia medica and ancient Chinese medical book, analyzed the local traditional Chinese medicine slices processing norms and modern pharmacopoeia. Meanwhile, we reviewed the history of PRA in function and processing, and discussed the modern processing technology, chemical composition and pharmacological research in some extent. We hope it can be helpful for some related research of PRA.

Orofacial antinociceptive activity and anchorage molecular mechanism in silico of geraniol

Abstract We aimed to evaluate the orofacial antinociceptive effect of geraniol in mice and its molecular anchorage mechanism. Seven mice per group (probabilistic sample) were treated with geraniol (12.5, 25 and 50 mg/kg, i.p.), morphine (6 mg/kg, i.p.) and vehicle (saline + Tween 80 at 0.2%, i.p.) 30 minutes prior to the beginning of the experiment. Injecting glutamate (25 μM), capsaicin (2.5 μg) and formalin (2%) into the right upper lip (perinasal) of the mouse induced nociception. Behavioral analysis of the animals considered the friction time (in seconds) of the mentioned region using hind or front paws by a researcher blinded to the treatment groups. The statistical analysis was performed blindly, considering α = 5%. The results showed that in the glutamate and capsaicin tests, concentrations of 25 mg/kg and 50 mg/kg presented antinociceptive activity (p < 0.005, power> 80%). In the formalin test, geraniol was able to reduce nociception at a concentration of 50 mg/kg (p < 0.005, power> 80%). In the molecular anchorage study, high values of binding between the evaluated substance and receptors of glutamate were observed (metabotropic glutamate receptor, -87.8501 Kcal/mol; N-methyl-D-aspartate, -86.4451 Kcal/mol; α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid, -85.6755 Kcal/mol). Geraniol presented orofacial antinociceptive activity, probably by interacting with glutamate-related receptors.

Bradycardic and Antiarrhythmic Effects of the D-Limonene in Rats / Efeitos Bradicárdicos e Antiarrítmicos do D-Limoneno em Ratos

Abstract Background: D-limonene (DL) is a monoterpene and is the major component in the essential oil of citrus fruit. It presents antihyperglycemic and vasodilatation activities. Objectives: This study evaluated the cardiovascular effects and potential antiarrhythmic of DL in rats. Methods: Hemodynamic and electrocardiographic (ECG) parameters were measured in male Wistar rats, which under anesthesia had been cannulated in the abdominal aorta and lower vena cava and had electrodes subcutaneously implanted. In the in vitro approach, the heart was removed and perfused using the Langendorff technique. The significance level adopted was 5% (p < 0.05). Results: DL, in doses of 10, 20, and 40 mg/kg (i.v), produced intense and persistent bradycardia associated with hypotension. Bradycardia with prolonged QTc was observed in the ECG in vivo recording. In the in vivo model of arrhythmia induced by Bay K8644, DL (10 mg/kg) decreased the arrhythmia score from 15.33 ± 3.52 to 4.0 ± 2.64 u.a (p < 0.05, n = 4). In isolated perfused hearts, DL (10-3 M) promoted significant reductions in heart rate (from 228.6 ± 8.5 ms to 196.0 ± 9.3 bpm; p < 0.05) and left ventricular development pressure (from 25.2 ± 3.4 to 5.9 ± 1.8 mmHg; n = 5, p < 0.05). Conclusions: DL produces bradycardia and antiarrhythmic activity in rat heart.

Resumo Fundamento: O D-limoneno (DL) é um monoterpeno e o principal componente do óleo essencial de frutas cítricas. Ele apresenta atividades anti-hiperglicêmicas e vasodilatadoras. Objetivos: Este estudo avaliou os efeitos cardiovasculares e antiarrítmicos potenciais do DL em ratos. Métodos: Os parâmetros hemodinâmicos e eletrocardiográficos (ECG) foram mensurados em ratos Wistar machos que, sob anestesia, tiveram a aorta abdominal e a veia cava inferior canuladas e receberam eletrodos implantados subcutaneamente. Na abordagem in vitro, o coração foi removido e perfundido utilizando a técnica de Langendorff. O nível de significância adotado foi de 5% (p < 0,05). Resultados: DL, nas doses de 10, 20 e 40 mg/kg (i.v), produziu bradicardia intensa e persistente associada à hipotensão. A bradicardia com QTc prolongado foi observada no registro in vivo do ECG. No modelo in vivo de arritmia induzida por Bay K8644, DL (10 mg / kg) houve diminuição do escore da arritmia de 15,33 ± 3,52 para 4,0 ± 2,64 u.a (p < 0,05, n = 4). Em corações perfundidos isolados, o DL (10-3 M) promoveu reduções significativas na frequência cardíaca (de 228,6 ± 8,5 ms para 196,0 ± 9,3 bpm; p < 0,05) e na pressão desenvolvida do ventrículo esquerdo (de 25,2 ± 3,4 para 5,9 ± 1,8 mmHg; n = 5, p < 0,05). Conclusões: O DL produz bradicardia e atividade antiarrítmica no coração de ratos.

Anti-inflammatory and anti-nociceptive activities of essential oil of Waltheria indica / Actividades antiinflamatorias y nociceptivas del aceite esencial de Waltheria indica

This paper reports for the first time volatile compounds, anti-nociceptive and anti-inflammatory activities of essential oils from the leaves of Waltheria indica L. (Stericullaceae) growing in Nigeria. The essential oil was hydro-distilled and characterized by gas chromatography-flame ionization detection (GC-FID) and gas chromatography coupled with mass spectrometry (GC-MS) analyses. The anti-inflammatory activity was evaluated on carrageenan induced rat paw edema while the anti-nociceptive test was based on hot plate model. The hydro-distillation afforded 0.41% (dry weight basis) of light green oil. Forty compounds representing 99.8% were identified in the oil. The main constituents of the oil were limonene (34.7%), sabinene (21.2%) and citronellal (9.7%). The anti-nociceptive property of the essential oils statically inhibited edema development (p<0.001) at a dose of 200 and 400 mg/kg independent of time of exposure. However, the 100 mg/kg Waltheria indica essential oils (WIEO) displayed a relatively low inhibition (p<0.01-p>0.5) which declines as exposure time increases. The anti-inflammatory activities shows a steady rate and non-dose dependent activity (p<0.001) up to the 3rd h of inflammation study. Conversely, a sharp reduction at the rate of p<0.5, 0.1 and 0.01 for the 100, 200 and 400 mg/kg WIEO doses respectively. Overall, the results presented sustain and establish the anti-nociceptive and anti-inflammatory properties and justifies the need for further evaluation and development of the essential oils from this plant.

Este artículo informa por primera vez de compuestos volátiles, actividades anti-nociceptivas y antiinflamatorias de aceites esenciales de las hojas de Waltheria indica L. (Stericullaceae) que crecen en Nigeria. El aceite esencial fue hidro-destilado y se caracterizó por cromatografía de gases-detección de ionización de llama (GC-FID) y cromatografía de gases junto con análisis de espectrometría de masas (GC-MS). La actividad antiinflamatoria se evaluó en el edema de pata de rata inducido por carragenano, mientras que la prueba antinociceptiva se basó en el modelo de placa caliente. La destilación hidráulica proporcionó 0,41% (en peso seco) de aceite verde claro. Cuarenta compuestos que representan el 99.8% fueron identificados en el aceite. Los principales componentes del aceite fueron el limoneno (34,7%), el sabineno (21,2%) y el citronelal (9,7%). La propiedad anti-nociceptiva de los aceites esenciales inhibió estáticamente el desarrollo del edema (p<0.001) a una dosis de 200 y 400 mg/kg independientemente del tiempo de exposición. Sin embargo, los aceites esenciales de Waltheria indica de 100 mg/kg (WIEO) mostraron una inhibición relativamente baja (p<0.01-p>0.5) que disminuye a medida que aumenta el tiempo de exposición. Las actividades antiinflamatorias muestran una tasa constante y una actividad no dependiente de la dosis (p<0.001) hasta la tercera hora del estudio de inflamación. Por el contrario, una fuerte reducción a una tasa de p<0.5, 0.1 y 0.01 para las dosis de 100, 200 y 400 mg/kg de WIEO respectivamente. En general, los resultados presentados sostienen y establecen las propiedades anti-nociceptivas y antiinflamatorias y justifican la necesidad de una mayor evaluación y desarrollo de los aceites esenciales de esta planta.

Research progress of chemical constituents and biological activities of essential oil of Pistacia lentiscus / 中国中药杂志

Pistacia lentiscus,which belongs to foreign medicine resources,is widely distributed in the Mediterranean and Middle Eastern area. The essential oils are a mixture of several volatile compounds mainly monoterpenes and sesquiterpenes obtained from different parts of P. lentiscus by hydrodistillation. The variability of chemical composition,biological activities and content of essential oil is strongly affected by extraction technology,environmental and sex factors. It is indicated that essential oils of P. lentiscus have kinds of biological activities such as antibacterial,anticancer,anti-atherogenesis,antioxidant,anti-inflammatory and insecticidal activities.Many scholars hold the opinion that combination of different components with synergistic and/or additive actions should account for their biological activities. Due to its diverse efficacy and special taste,the essential oil of P. lentiscus has been extensively used in medicine,food and cosmetics industries. A mini review of chemical constituents and biological activities of essential oil of P. lentiscus in the past20 years is made here to provide valuable reference for the construction of " the Belt and Road".

Essential oils of Myrcia lundiana Kiaersk and their major compounds show differentiated activities against three phytopathogenic fungi / Óleos essenciais de Myrcia lundiana Kiaersk e seus compostos majoritários apresentam atividades diferenciadas sobre três fungos fitopatogênicos

The present study evaluated the antifungal activity of the essential oils of chemotypes of Myrcia lundiana and their major compounds on the fungi Fusarium pallidoroseum, Fusarium solani, and Colletotrichum musae. The essential oils were obtained by hydrodistillation and analyzed by GCMS/FID. For the evaluation of the antifungal activity, the essential oils and the major compounds were tested at the concentration of 0.1 mL/L until the fungicidal effect was detected. The major compounds detected in the essential oil were 1,8-cineole, isopulegol, and citral. The chemotypes (MLU-005 and MLU-019) provided 100% mycelial growth inhibition for the fungus F. pallidoroseum from the concentration of 1.1 mL/L (minimum inhibition concentration - MIC). For chemotype (MLU-022), the minimum fungicidal concentration (MFC) was 0.3 mL/L. For F. solani, the essential oils of the chemotypes (MLU-005 and MLU-019) presented MIC at concentrations of 7.0 and 5.0 mL/L, respectively. The essential oil of the chemotype (MLU-022) presented MFC of 0.6 mL/L. Different MIC was observed for the three studied chemotypes for the fungus C. musae, ranging between 0.4 mL/L, for the chemotype (MLU-005); 0.5 mL/L, for the chemotype (MLU-022); and 0.7 mL/L, for the chemotype (MLU-019). The best MFC was observed for the chemotype (MLU-005) (0.5 mL/L). The major compounds tested separately presented better MIC values when compared with their chemotypes, except for the compound 1,8-cineole, which presented lower mycelial growth inhibition for the three fungi tested, suggesting that the chemical profile or the presence of some other compound of the essential oil may inhibit the growth of the three fungi studied. The compound isopulegol provided lower MFC for the fungus C. musae (0.4517 mL/L) when compared with the fungi F. pallidoroseum and F. solani, (MFC of 0.4927 mL/L). The compound citral provided a lower MFC on the fungus C. musae (0.1668 mL/L) in relation to the other fungi tested. The essential oils of the chemotypes of M. lundiana and their major compounds showed potential to control the studied phytopathogens and can be an alternative for agriculture for presenting an inhibitory and fungicidal effect against these organisms at lower concentrations.

O presente trabalho avaliou a atividade antifúngica de óleos essenciais de quimiotipos de Myrcia lundiana dos seus compostos majoritários sobre os fungos Fusarium pallidoroseum, Fusarium solani e Colletotrichum musae. Os óleos essenciais foram obtidos por hidrodestilação e analisados por CGEM/DIC. Para avaliação da atividade antifúngica, foram testados os óleos essenciais e os compostos majoritários na concentração de 0,1 mL/L até encontrar o efeito fungicida. Os principais compostos presentes no óleo essencial foram 1,8-cineol, isopulegol e citral. Os quimiotipos (MLU-005 e MLU-019) proporcionaram 100% de inibição do crescimento micelial para o fungo F. pallidoroseum a partir da concentração de 1,1 mL/L (Concentração Inibitória Mínima ­ CIM). Para o quimiotipo (MLU-022), a melhor concentração fungicida mínima (CFM) foi de 0,3 mL/L. Para F. solani, os óleos essenciais dos quimiotipos (MLU-005 e MLU-019) apresentaram CIM nas concentrações de 7,0 e 5,0 mL/L, respectivamente. O óleo essencial do quimiotipo (MLU-022) apresentou CFM de 0,6 mL/L. Observou-se CIM diferenciado para os três quimiotipos estudados para o fungo C. musae, variando entre 0,4 mL/L, para o quimiotipo (MLU-005); 0,5 mL/L, para o quimiotipo (MLU-022); e 0,7 mL/L, para o quimiotipo (MLU-019). O quimiotipo MLU-005 apresentou o melhor CFM, 0,5 mL/L. Os compostos majoritários testados separadamente apresentaram melhores valores de CIM frente aos seus quimiotipos, exceto o composto 1,8-cineol, que apresentou menor inibição do crescimento micelial para os três fungos testados, sugerindo que o perfil químico ou a presença de algum outro composto no óleo essencial pode estar atuando na inibição do crescimento dos três fungos estudados. O composto isopulegol proporcionou menor CFM para o fungo C. musae (0,4517 mL/L) em relação aos fungos F. pallidoroseum e F. solani, para os quais apresentou CFM de 0,4927 mL/L. O composto citral proporcionou um menor CFM sobre o fungo C. musae (0,1668 mL/L), em relação aos demais fungos testados. Os óleos essenciais de quimiotipos de M. lundiana e seus compostos majoritários apresentaram potencial para o controle dos fitopatógenos estudados, podendo ser considerados como uma alternativa para a agricultura, uma vez que em concentrações mais baixas apresentaram efeito inibitório e fungicida frente a estes organismos.

Influence of growth regulators on distribution of trichomes and the production of volatiles in micropropagated plants of Plectranthus ornatus

ABSTRACT The profile of volatile organic compounds, the glandular and non-glandular trichomes of Plectranthus ornatus, obtained by in vitro cultivation, was evaluated in plants grown in Murashide and Skoog medium supplemented with benylaminopurine at 4.5, 9.0, and 18.0 µM + naphthaleneacetic acid at 5.37 µM, kinetin at 4.7, 9.3 and 18.5 µM + naphthaleneacetic acid (5.37 µM) or Murashide and Skoog 0 medium (as a control). Scanning Electron Microscopy was performed on samples of the third leaf node of the 90 days old plants obtained from treatment with 4.5 or 9.0 µM benylaminopurine, and 4.7 or 9.3 µM kinetin. Headspace Solid Phase Micro-Extraction of the 30, 60 and 90 days old in vitro plants permitted to determinate by GC/MS the composition comprised of 62 compounds. The data were analyzed using Principal Component Analysis and Hierarchical Clustering Analysis and, the major constituents of these oils after treatment and aging were monoterpenes and sesquiterpenes. Morphoanatomical analysis of trichomes, by Scanning Electron Microscopy, enabled the identification of non-glandular trichomes and four types of glandular trichomes, which comprised capitate and peltate glandular trichomes that were distributed on both sides of the leaf. We observed that the regulators influenced qualitative and quantitative profiles of the volatile organic compounds and the number and distribution of hairs on the leaf surface.