In vitro antibacterial-antibiofilm effect of Hypericum atomarium Boiss and chemical composition

Abstract Treatment with plant is considered an effective option against increased antibiotic resistance. In this study antibiofilm activity of methanol (CH3OH), chloroform (CHCl3), ethyl acetate (EtOAc) and water (H2O) extracts of Hypericum atomarium Boiss. which is member of Hypericum genus was evaluated in Pseudomonas aeruginosa PAO1 and antibacterial performance against Gram (+) and Gram (-) strains and also bioactive compounds of extract were analysed using by HPLC and GC-MS. According to antibacterial activity test results the extracts were effective all Gram (+) bacteria and Gram (-) Chromobacterium violaceum (MICs ranging from 0.42 µg/ml to 4.3 mg). Inhibition effect of biofilm formation was found to be different rate in extracts (methanol-63%, chloroform-52%). The major flavonoids were detected (−)-epicatechin (2388.93 µg/ml) and (+)-catechin (788.94 µg/ml). The main phenolic acids were appeared as caffeic acid 277.34 µg/ml and chlorogenic acid 261.79 µg/ml. And according to GC results α-pinene was found main compound for three solvent extracts methanol, chloroform and ethyl acetate 67.05, 62.69, 49.28% rate respectively

Establecimiento in vitro de Hypericum goyanesii Cuatrec. E Hypericum juniperinum Kunth, a partir del cultivo de semillas / Establishing in vitroHypericum goyanesii Cuatrec. E Hypericum juniperinum Kunth, from seeds culture

El propósito de este estudio fue evaluar medios de cultivo de germinación para el establecimiento in vitro de Hypericum goyanesii e Hypericum juniperinum como estrategia de conservación ex situ, por medios biotecnológicos de especies vegetales pertenecientes al ecosistema alto-andino. Las semillas se desinfectaron con hipoclorito de sodio y se sembraron en seis tratamientos basados en los medios de cultivo Murashige & Skoog (MS) y MS con reducción del 50% de sus macro y microsales y vitaminas, con y sin la adición de carbón activado y pulpa de banano. La germinación se favoreció por el empleo de pulpa de banano en el medio MS con reducción de sales y suplementado con carbón activado. Fue posible evidenciar que los explantes sexuales de H. goyanesii e H. juniperinum estuvieron influenciados positivamente en la variable porcentaje de germinación, por el empleo de sustancias orgánicas en reemplazo de reguladores de crecimiento.

The purpose of this study was to evaluate germination culture media for the in vitro establishment Hypericum goyanesii e Hypericum juniperinum as ex situ conservation strategy for biotechnology of plant species belonging to the high-Andean ecosystem. The seeds were disinfected with sodium hypochlorite and seeded into six treatments MS and MS with 50% reduction of its macro and microsales and vitaminas, with and without the addition of activated charcoal and pulp banana. Germination is favored by the use of banana pulp in MS medium with reducing salts and supplemented with activated charcoal. It was possible to show that sexual explants and H. goyanesii e H. goyanesii juniperinum were influenced positively variable germination percentage, by the use of organic substances to replace growth regulators.

Composición química y evaluación de la actividad antifúngica del aceite esencial de corteza de Vismia baccifera var. dealbata / Chemical composition and evaluation of anti-fungal activity of essential oil from Vismia baccifera var. dealbata bark

El objetivo de este trabajo fue evaluar la susceptibilidad de levaduras del género Candida y el Complejo Cryptococcus neoformans al aceite esencial de Vismia baccifera var. dealbata. El aceite, procedente de Chiguará, estado Mérida-Venezuela, fue analizado por CG/ EM logrando la identificación de trece componentes, que constituyeron el 97,7% de la mezcla; tres de ellos se apreciaron como productos mayoritarios, representando el 70,4% de la totalidad (Óxido de cariofileno 31,4%, β-cariofileno 26,4% y α-zingibireno 12,6%). El ensayo de actividad antifúngica mostró que dicho aceite inhibió el crecimiento de varias cepas de los géneros Candida y el Complejo C. neoformans, evaluadas cualitativamente por difusión en agar con disco a una concentración de 1.000 μg/mL; se observaron halos de inhibición entre 8 y 12 mm, exhibiendo valores de CMI entre 1,6 y 1.000 μg/mL en el ensayo cuantitativo por el método “Spot on a lawn”. Los datos obtenidos fueron sometidos a análisis de varianza y la prueba de Tukey, obteniéndose una actividad inhibitoria relevante contra Candida krusei. Este estudio constituye el primer reporte, tanto de la composición química como de evaluación antifúngica del aceite esencial extraído a partir de especies del género Vismia.

The purpose of this study was to evaluate the susceptibility of Candida genus yeasts and the Cryptococcus neoformans Complex to essential oil from Vismia baccifera var. dealbata. This oil, obtained at Chiguará, Mérida State, was analyzed by GC/MS, which identified thirteen components that constituted 97.7% of the mixture; three of them were established as majority products, representing 70.4% of the total: (cariophyllene oxide 31.4%, β-cariophyllene 26.4%, and α-zingibirene 12.6%). The anti-fungal assay showed that this oil inhibited the growth of several Candida genus strains and of the C. neoformans Complex, qualitatively evaluated by agar disk diffusion at a 1,000 μg/ mL concentration; inhibition halos of a between 8 to 12 mm diameter were observed, showing MIC values between 1.6 and 1,000 μg/mL in the “Spot on a lawn” quantitative assay. The data obtained were submitted to ANOVA and Tukey’s test, demonstrating a relevant inhibitory activity against Candida krusei. This study constitutes the first report, both of the chemical composition and of the anti-fungal evaluation, of the essential oil extracted from Vismia genus species.

Hypericum caprifoliatum and Hypericum connatum affect human trophoblast-like cells differentiation and Ca2+influx

Objective:To study the effect of crude methanol and n-hexane extracts of Hypericum connatum (H. connatum) and Hypericum caprifoliatum on trophoblast-like cells. Methods: BeWo and JEG-3 trophoblast-like cells were submitted to different extract concentrations (1, 5, 10 and 15 μg/mL) and evaluated in relation to cell viability and in vitro trophoblast differentiation and function. Cell viability was evaluated using WST-1 reagent. Differentiation was measured by luciferase production, hCG production/release, and mitogen-activated protein kinase signaling pathway activation. The function of the trophoblast-like cells was measured by 45Ca2+influx evaluation. Results:The results showed a decrease in cell viability/proliferation. Both plants and different extracts induced a significant decrease in hCG production/release and luciferase production. H. connatum did not cause mitogen-activated protein kinase signaling pathway disturbance;however, Hypericum caprifoliatum n-hexane extract at 15 μg/mL inhibited extracellular signal-regulated kinase 1/2 activation. The significant increase in Ca2+influx by JEG-3 cells was seen after short and long incubation times with H. connatum methanolic extract at 15 μg/mL. Conclusions: The results indicated that these two Hypericum species extracts can interfere on trophoblast differentiation and Ca2+influx, according to their molecular diversity. Although in vivo experiments are necessary to establish their action on placental formation and function, this study suggests that attention must be paid to the potential toxic effect of these plants.

Hypericum caprifoliatum and Hypericum connatum affect human trophoblast-like cells differentiation and Ca

Objective: To study the effect of crude methanol and n-hexane extracts of Hypericum connatum (H. connatum) and Hypericum caprifoliatum on trophoblast-like cells. Methods: BeWo and JEG-3 trophoblast-like cells were submitted to different extract concentrations (1, 5, 10 and 15 μg/mL) and evaluated in relation to cell viability and in vitro trophoblast differentiation and function. Cell viability was evaluated using WST-1 reagent. Differentiation was measured by luciferase production, hCG production/release, and mitogen-activated protein kinase signaling pathway activation. The function of the trophoblast-like cells was measured by

Hypericum caprifoliatum and Hypericum connatum affect human trophoblast-like cells differentiation and Ca(2+) influx

<p><b>OBJECTIVE</b>To study the effect of crude methanol and n-hexane extracts of Hypericum connatum (H. connatum) and Hypericum caprifoliatum on trophoblast-like cells.</p><p><b>METHODS</b>BeWo and JEG-3 trophoblast-like cells were submitted to different extract concentrations (1, 5, 10 and 15 µg/mL) and evaluated in relation to cell viability and in vitro trophoblast differentiation and function. Cell viability was evaluated using WST-1 reagent. Differentiation was measured by luciferase production, hCG production/release, and mitogen-activated protein kinase signaling pathway activation. The function of the trophoblast-like cells was measured by (45)Ca(2+) influx evaluation.</p><p><b>RESULTS</b>The results showed a decrease in cell viability/proliferation. Both plants and different extracts induced a significant decrease in hCG production/release and luciferase production. H. connatum did not cause mitogen-activated protein kinase signaling pathway disturbance; however, Hypericum caprifoliatum n-hexane extract at 15 µg/mL inhibited extracellular signal-regulated kinase 1/2 activation. The significant increase in Ca(2+) influx by JEG-3 cells was seen after short and long incubation times with H. connatum methanolic extract at 15 µg/mL.</p><p><b>CONCLUSIONS</b>The results indicated that these two Hypericum species extracts can interfere on trophoblast differentiation and Ca(2+) influx, according to their molecular diversity. Although in vivo experiments are necessary to establish their action on placental formation and function, this study suggests that attention must be paid to the potential toxic effect of these plants.</p>

Protective effect of Hypericum calabricum Sprengel on oxidative damage and its inhibition of nitric oxide in lipopolysaccharide-stimulated RAW 264.7 macrophages

The present study shows for the first time the phenolic composition and the in vitro properties (antioxidant and inhibition of nitric oxide production) of Hypericum calabricum Sprengel collected in Italy. The content of hypericins (hypericin and pseudohypericin), hyperforin, flavonoids (rutin, hyperoside, isoquercetrin, quercitrin, quercetin and biapigenin) and chlorogenic acid of H. calabricum, have been determined. The ethyl acetate fraction from the aerial parts of H. calabricum exhibited activity against the radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) with IC50 value of 1.6 jig/ml. The test for inhibition of nitric oxide (NO) production was performed using the murine monocytic macrophage cell line RAW 264.7. The ethyl acetate fraction had significant activity with an IC50 value of 102 jig/ml and this might indicate that it would have an anti-inflammatory effect in vivo.

Influence of an aqueous extract of Hypericum perforatum (Hypericin) on the survival of Escherichia coli AB1157 and on the electrophoretic mobility of pBSK plasmid DNA / Influência de um extrato aquoso de Hypericum perforatum (Hipericina) na sobreviência de Escherichia coli AB1157 e na mobilidade eletroforética de DNA plasmidial pBSK

Hiperico (Hypericum perforatum or St John's worth) has been widely used as an herbal medicine to treat depression. Hypericin is the main chemical compound of hiperico. Stannous chloride (SnCl2) is the most used reducing agent in nuclear medicine. The aim of this work was to verify the effect of a hiperico extract on the survival of Escherichia coli AB1157 and on the plasmid DNA topology. Exponentially E. coli AB1157 cultures were incubated with SnCl2 in the presence or absence of hypericin. Aliquots were spread onto Petri dishes containing solidified rich medium, the colonies units were counted after overnight and the survival fraction was calculated. Plasmid DNA samples were incubated with SnCl2 in presence or absence of hypericin extract during 40 minutes, 0.8 percent agarose gel electrophoresis was performed, the gel was stained with ethidium bromide and the plasmid topological forms (bands) were visualized. The results revealed that hiperico extract is neither capable of altering the survival of E. coli cells nor the plasmid DNA topology but it may have protected these cells against the SnCl2 action. The data suggest absence of cytotoxic and genotoxic effects of the aqueous hiperico extract and a protective effect on E. coli cells against the action of SnCl2.

Hipérico (Hypericum perforatum or St John's worth) tem sido usado como uma planta medicinal para tratar a depressão. Hipericina é o principal componente do hipérico. O cloreto estanoso (SnCl2) é o agente redutor mais utilizado em medicina nuclear. O objetivo desse trabalho foi verificar o efeito de um extrato de hipérico na sobrevivência de Escherichia coli AB1157 e na topologia do DNA plasmidial. Culturas de E. coli AB1157, em fase exponencial, foram incubadas com SnCl2 na presença ou ausência de hipericina. Alíquotas foram espalhadas em placas de Petri contendo meio sólido, as unidades formadoras de colônias foram contadas após incubação e as frações de sobrevivência calculadas. DNA plasmidial foi incubado com SnCl2 na presença ou ausência de hipericina durante 40 minutos, eletroforese em gel de agarose a 0,8 por cento foi realizada, o gel foi corado com brometo de etídio e as formas (bandas) topológicas do plasmídeo visualizadas. Os resultados revelaram que o extrato de hipérico não foi capaz de alterar a sobrevivência da cultura de E. coli e a topologia do DNA plasmidial, mas protegeu as bactérias contra a ação do SnCl2. Os resultados sugerem ausência de efeitos citotóxicos e genotóxicos do extrato aquoso do hipérico e um efeito protetor nas células de E. coli contra a ação do SnCl2.