Influence of Sucrose and Activated Charcoal on Phytochemistry and Vegetative Growth in Zephyranthes irwiniana (Ravenna) Nic. García (Amaryllidaceae).

Zephyranthes irwiniana (Ravenna) Nic. García is an endemic, red list threatened species from the Brazilian savanna (Cerrado) with pharmacological potential to treat the symptoms of Alzheimer's Disease (AD). This work analyzed the vegetative growth and phytochemistry of its potential compounds, in response to variations in sucrose concentration and activated carbon (AC). Seeds were germinated in vitro and in the greenhouse. The in vitro bulbs were separated in six treatments with different sucrose concentrations (30, 45 and 60 gL-1) and/or AC (1 gL-1). Biomass increases in individuals grown in the greenhouse were higher than those cultivated in vitro. Sucrose concentration significantly increased biomass and root number. AC had a positive influence on leaf and root size, and a negative influence on root number. GC-MS analyses indicated great variation in the abundance of α-terpenyl-acetate, ethyl linoleate, clionasterol and lycorine between treatments, with maximum concentrations of 53.06%, 38.68, 14.34% and 2.57%, respectively. Histolocalization tests indicated the presence of alkaloids in the leaf chlorenchyma and bulb cataphylls. Finally, the present study provided new evidence that the constitution of the culture medium directly influences the vegetative growth and phytochemistry of this species, providing a good medium condition for propagating the species under threat.

Leaf anatomy and photosynthetic parameters of Vellozia squamata Pohl (Velloziaceae) grown under different light intensities along in vitro cultivation / Anatomia foliar e parâmetros fotossintéticos de Vellozia squamata Pohl (Velloziaceae) desenvolvendo-se sob diferentes intensidades luminosas ao longo do cultivo in vitro

The present work evaluates photosynthetic parameters and the leaf anatomy of plants of Vellozia squamata Pohl (Velloziaceae) in each phase of in vitro production, with wild plants serving as a control. The anatomical approach was done for the leaf middle portion. The photosynthetic curves were obtained with an infra-red gas analyzer. The in vitro plantlets showed the thinner and poorest developed leaves, the wild plants had the thickest and most developed leaves, and the plants in acclimatization showed intermediate features. The physiological pattern was similar, and the in vitro plantlets were not capable of net carbon absorption. The young seedlings at the garden showed a maximal net carbon assimilation rate inferior to the wild plants. In conclusion, the light intensity for the in vitro phase should be adjusted to produce seedlings.

O presente trabalho avalia parâmetros fotossintéticos e a anatomia foliar de Vellozia squamata Pohl (Velloziaceae) em cada etapa da produção in vitro, sendo as plantas silvestres o grupo-controle. A abordagem anatômica foi feita para a porção mediana foliar. As curvas fotossintéticas foram obtidas com analisador de gases por infravermelho. As plântulas in vitro apresentaram folhas mais delgadas e menos desenvolvidas, as silvestres apresentaram folhas mais espessas e desenvolvidas e aquelas em aclimatação exibiram características intermediárias. As plântulas in vitro não foram capazes de absorção líquida de carbono. As plântulas jovens de casa-de-vegetação exibiram taxa líquida de assimilação de carbono inferior às selvagens. Em conclusão, para o cultivo in vitro as intensidades luminosas para a produção de mudas devem ser melhor ajustadas.

Wound Healing Effect of Essential Oil Extracted from Eugenia dysenterica DC (Myrtaceae) Leaves.

The use of natural oils in topical pharmaceutical preparations has usually presented safe agents for the improvement of human health. Based on research into the immense potential of wound management and healing, we aimed to validate the use of topical natural products by studying the ability of the essential oil of Eugenia dysenterica DC leaves (oEd) to stimulate in vitro skin cell migration. Skin cytotoxicity was evaluated using a fibroblast cell line (L929) by MTT assay. The oil chemical profile was investigated by GC-MS. Moreover, the inhibition of lipopolysaccharide (LPS) induced nitric oxide (NO) production in the macrophage cell line (RAW 264.7) tested. The Chick Chorioallantoic Membrane (CAM) assay was used to evaluate the angiogenic activity and irritating potential of the oil. The oEd induces skin cell migration in a scratch assay at a concentration of 542.2 µg/mL. α-humulene and ß-caryophyllene, the major compounds of this oil, as determined by GC-MS, may partly explain the migration effect. The inhibition of nitric oxide by oEd and α-humulene suggested an anti-inflammatory effect. The CAM assay showed that treatment with oEd ≤ 292 µg/mL did not cause skin injury, and that it can promote angiogenesis in vivo. Hence, these results indicate the feasibility of the essential oil of Eugenia dysenterica DC leaves to developed dermatological products capable of helping the body to repair damaged tissue.

Activity of crude extracts from Brazilian cerrado plants against clinically relevant Candida species.

BACKGROUND: Medicinal plants have traditionally been used in many parts of the world as alternative medicine. Many extracts and essential oils isolated from plants have disclosed biological activity, justifying the investigation of their potential antimicrobial activity. In this study, the in vitro antifungal activity of six Brazilian Cerrado medicinal plant species were evaluated against clinically relevant Candida species. METHODS: The crude extract plants were evaluated against American Type Culture Collection (ATCC) standard strains of Candida spp. using disk diffusion method and determining the minimum inhibitory concentration (MIC). The chemical study results were confirmed by HPLC method. RESULTS: All six plant species showed antifungal activity. Among the species studied, Eugenia dysenterica and Pouteria ramiflora showed significant inhibitory activity against C. tropicalis at lowest MIC value of 125 and 500 µg/disc, respectively. The Eugenia dysenterica also disclosed MIC value of 125 µg/disc against C. famata, 250 µg/disc against C. krusei and 500 µg/disc against C. guilliermondii and C. parapsilosis. Pouteria torta, Bauhinia rufa, Erythroxylum daphnites and Erythroxylum subrotundum showed activity against the yeast strains with MIC value of 1000 µg/disc. The chemical study of the most bioactive extracts of Eugenia dysenterica and Pouteria ramiflora revealed catechin derivatives and flavonoids as main components. CONCLUSIONS: All six evaluated plant species showed good antifungal potential against several Candida strains. However, E .dysenterica and P. ramiflora showed the higher inhibitory effect against the non-albicans Candida species. Our results may contribute to the continuing search of new natural occurring products with antifungal activity.

Determination of rutin in Erythroxylum suberosum extract by liquid chromatography: applicability in standardization of herbs and stability studies / Determinación de rutina en el extracto Erythroxylum suberosum por cromatografía líquida: aplicación de la normalización de las hierbas y los estudios de estabilidad

In this study, a reverse phase-high performance liquid chromatography (RP-HPLC) technique for determination of rutin in Erythroxylum suberosum extract was developed and validated. A regression analysis was performed, with the observation of good linearity (r = 0.9997). The values obtained for precision and accuracy determination are in agreement with ICH guidelines. The detection and the quantitation limits of rutin were 0.19 ug/mL and 0.60 ug/mL, respectively. The results demonstrated that the developed method is a reliable HPLC technique for determination of rutin in E. suberosum extract. In addition, the applicability of this method in stability studies and standardization of herbs was investigated.

En este estudio, la técnica de cromatografía líquida de alta resolución en fase reversa para la determinación de la rutina en el extracto Erythroxylum suberosum fue desarrollada y validada. Se realizó un análisis de regresión, con la observación de una buena linealidad (r = 0,9997). Los valores obtenidos para la precisión y la determinación de la precisión están de acuerdo con las directrices ICH. La detección y cuantificación de los límites de la rutina fueron 0,19 ug / mL y 0,60 ug / mL, respectivamente. Los resultados demostraron que el método desarrollado es una técnica fiable de HPLC para la determinación de la rutina en el extracto de E. suberosum. Además, se investigó la aplicabilidad de este método en los estudios de estabilidad y la estandarización de hierbas.

Anxiogenic effects of Tabernaemontana solanifolia A. DC. (Apocynaceae) extracts in rats tested in the elevated plus-maze / Determinación mediante la prueba del laberinto elevado en cruz del efecto ansiogénico en ratas de extractos de Tabernaemontana solanifolia A. DC. (Apocynaceae)

Introduction: Tabernaemontana solanifolia A. DC. (Apocynaceae) grows in the Cerrado (savanna), Caatinga (shrublands), and Atlantic Forest of Brazil. Objective: to investigate the potential anxiolytic activity of the crude extracts and ß-amyrin acetate obtained from the leaves of Tabernaemontana solanifolia in the elevated plus-maze (EPM) test for anxiety in rats. Methods: the crude extracts (aqueous, ethanol, and hexane), as well as ß-amyrin acetate, obtained from Tabernaemontana solanifolia leaves were evaluated for possible anxiolytic effects in rats tested in the elevated plus-maze. Acute toxicity in rats was determined by OECD 423 guidelines. The leaves of T. solanifolia specimens collected in Brasília, Brazil, were air dried at 40 °C and macerated with hexane or ethanol. After filtration, the solvents were removed under reduced pressure, yielding the crude hexane and ethanolic extracts (5.96 and 18.62 % yield, respectively). The crude aqueous extract was obtained by infusion, followed by lyophilization (13.5 % yield). Thirty minutes before the elevated plus-maze experiments, the animals were treated with the crude ethanol extract (1 000 mg/kg of body weight [bw], p.o.), the crude hexane extract (1 000 mg/kg bw, p.o.), the crude extract (1 000 mg/kg bw, p.o.), or ß-amyrin acetate (21 mg/kg bw, p.o.). Results: the extracts cauded no mortality up to 2 000 mg/kg, so half of DL50 doses were selected for the present study. The crude extracts (hexane, ethanolic and aqueous) (1.0 g/kg) as well as the substance ß-amyrin acetate (21 mg/kg) were administered once, increasing the number of entries into and the time spent in the closed arms of the elevated plus-maze. Conclusion: the oral administration of the extracts and ß-amyrin acetate obtained from Tabernaemontana solanifolia had an anxiogenic effect in rats. Further studies are needed in order to identify and characterize the anxiogenic properties of Tabernaemontana solanifolia and to understand the anxiogenic mechanisms of the extracts.

Introducción: Tabernaemontana solanifolia A. DC. (Apocynaceae) crece en el Cerrado (sabana), Caatinga (matorrales) y la Mata Atlántica de Brasil. Objetivo: investigar la posible actividad ansiolítica de los extractos crudos y el acetato de ß-amirina obtenidos de las hojas de Tabernaemontana solanifolia en la prueba del laberinto elevado en cruz (EPM) para la ansiedad en ratas. Métodos: los extractos crudos (etanol acuoso y hexano), así como el acetato de ß-amirina, se evaluaron para posibles efectos ansiolíticos en ratas mediante la prueba del laberinto elevado en cruz. La toxicidad aguda en ratas se determinó por las directrices OCDE 423. Las hojas de Tabernaemontana solanifolia se secaron al aire a 40 °C y fueron maceradas con hexano y etanol. Después de la filtración, los disolventes se eliminaron bajo presión reducida, produciendo los extractos crudos hexano y etanólico (5,96 y 18,62 % de rendimiento, respectivamente). El extracto acuoso bruto se obtuvo por infusión, seguida por liofilización (13,5 % de rendimiento). A 30 min antes de los experimentos de laberinto elevado en cruz, los animales se trataron con el extracto de etanol crudo (1 000 mg/kg de peso corporal [bw], p.o.), el extracto de hexano crudo (1 000 mg/kg de peso corporal, por vía oral), el extracto acuoso (1 000 mg/kg de peso corporal, por vía oral) o ß-amirina acetato (21 mg/kg de peso corporal, por vía oral). Resultados: los extractos no produjeron mortalidad hasta 2 000 mg/kg, por lo que para el presente estudio se seleccionó la mitad de la dosis DL50. Los extractos crudos (hexano, etanol y acuoso) (1 000 mg/kg), así como la sustancia acetato de ß-amirina (21 mg/kg) se administraron una vez, aumentando el número de entradas y el tiempo pasado en los brazos cerrados de la prueba del laberinto elevado en cruz. Conclusión: la administración oral de los extractos y el acetato de ß-amirina obtenidos de Tabernaemontana solanifolia tuvo un efecto ansiogénico en ratas. Se necesitan estudios adicionales para identificar y caracterizar las propiedades ansiogénicas de Tabernaemontana solanifolia y para entender los mecanismos ansiogénicos de los extractos.

Plants from Brazilian Cerrado with potent tyrosinase inhibitory activity.

The increased amount of melanin leads to skin disorders such as age spots, freckles, melasma and malignant melanoma. Tyrosinase is known to be the key enzyme in melanin production. Plants and their extracts are inexpensive and rich resources of active compounds that can be utilized to inhibit tyrosinase as well as can be used for the treatment of dermatological disorders associated with melanin hyperpigmentation. Using in vitro tyrosinase inhibitory activity assay, extracts from 13 plant species from Brazilian Cerrado were evaluated. The results showed that Pouteria torta and Eugenia dysenterica extracts presented potent in vitro tyrosinase inhibition compared to positive control kojic acid. Ethanol extract of Eugenia dysenterica leaves showed significant (p<0.05) tyrosinase inhibitory activity exhibiting the IC50 value of 11.88 µg/mL, compared to kojic acid (IC50 value of 13.14 µg/mL). Pouteria torta aqueous extract leaves also showed significant inhibitory activity with IC50 value of 30.01 µg/mL. These results indicate that Pouteria torta and Eugenia dysenterica extracts and their isolated constituents are promising agents for skin-whitening or antimelanogenesis formulations.