Berberine-fluconazole microparticle-based combination therapy to treat candidiasis infections.

AIM: This study aims to incorporate alginate microparticles containing berberine and fluconazole into two different types of pharmaceutical formulations, to subsequently evaluate the antifungal activity against Candida albicans. METHODS AND RESULTS: Alginate microparticles containing BBR (berberine) and FLU (fluconazole) were produced by the spray-drying technique, characterized and incorporated in two pharmaceutical formulations, a vaginal cream and artificial saliva. Broth microdilution, checkerboard, time-kill curve, and scanning electron microscopy were carried out to determine the antifungal effects of BBR and FLU against C. albicans. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of free BBR were 125 µg ml-1. Synergism between BBR and FLU was demonstrated by a fractional inhibitory concentration index (FICI) = 0.0762. The time-kill curve for the combination BBR + FLU showed a more pronounced decrease in fungal growth in comparison to free drugs, and an antibiofilm effect of BBR occurred in the formation and preformed biofilm. CONCLUSION: Alginate microparticles containing BBR and FLU were obtained and incorporated in a vaginal cream and artificial saliva. Both formulations showed good stability, antifungal effects, and organoleptic characteristics, which suggest that BBR-FLU microparticles in formulations have potential as antifungal therapy.

Development, characterization and anti - Mycobacterium tuberculosis activity of solid lipid nanoparticles for oral Piper corcovadensis roots extract delivery / Desarrollo, caracterización y actividad anti - Mycobacterium tuberculosis de nanopartículas lipídicas sólidas para la administración oral de extracto de raíces de Piper corcovadensis

The present study thus aimed at the development and physicochemical characterization of solid lipid nanoparticles loaded with crude extract of Piper corcovadensis roots (SLN - CEPc) and chitosan - coated solid lipid nanoparticles loaded with crude extract of P. corcovadensis roots (C - SLN - CEPc), as well as the determination of its antimycobacterial activity against Mycobacterium tuberculosis H37Rv, its cytotoxicity against the Vero cell line and evaluation in the hemolysis assay. Both formulat ions containing the encapsulated extract showed high encapsulation efficiency, formed by a monodispersed system with small and spherical particles, and there was no aggregation of particles. In the biological assays, SLN - CEPc and C - SLN - CEPc showed promisin g anti - M. tuberculosis activity with a minimum inhibitory concentration (MIC) of 12.5 µg/mL, whereas the cytotoxic concentrations obtained at 50% (CC 50 ) in Vero cells were 60.0 and 70.0 µg/mL, respectively. Therefore, nanoencapsulation showed satisfactory results, justifying its usage in the development of new products.

El presente estudio apuntó al desarrollo y caracterización fisicoquímica de na nopartículas lípidas en estado sólido, cargadas con extracto crudo de raíz de Piper c orcovadensis (SLN - CEPc) y nanopartículas lípidas en estado sólido cubiertas con quitosano cargadas co n extracto crudo de raíz de P. corcovadensis (C - SLN - CEPc), así como la determinación de su actividad antimico bacterial contra Mycobacterium tuberculosis H37Rv, su citotoxicidad contra la línea celular Vero y su evaluación en ensayo de hemólisis. Ambas formulaciones que contenían el extracto encapsulado mostraron alta eficien cia de encapsulación, formado por un sistema monodispersado con pequeñas partículas esféricas, y no hubo agregación de partículas. En los ensayos biológicos, SLN - CEPc y C - SLN - CEPc mostraron un a prometedora actividad anti - M. tuberculosis con una mínima conc entración inhibitoria (MIC) de 12,5 µg/mL, mientras que las concentraciones citotóxicas obtenidas al 50% (CC 50 ) en células Vero estuvo en 60,0 y 70,0 µg/mL, respectivamente. Por lo tanto, la nanoencapsulación mostró resultados satisfactorios, justificando su uso en el desarrollo de nuevos productos.

Valorization of Campomanesia xanthocarpa leaves: chemical composition and antioxidant activity of crude extract and fractions / Valorización de hojas de Campomanesia xanthocarpa: composición química y actividad antioxidante de extracto crudo y fracciones

Campomanesia xanthocarpa leaves are a byproduct of fruit production without studies on antioxidant activity. Thus, this study aimed to identify the antioxidant compounds of C. xanthocarpaleaves by ultra-high performance liquid chromatography coupled with electrospray ionization-quadrupole-time of flight-mass spectrometry (UHPLC-ESI/qTOF) and by different in vitro antioxidant methods. The crude extract of C. xanthocarpa leaves had a yield of 15.2% and only five out of 37 fractions of the crude extract had antioxidant activity. The crude extract presented greater antioxidant activity than the isolated fractions. The identified antioxidant compounds were phenolic acids (gallic acid and chlorogenic acid), flavonoids (quercetin and naringenin 7,4'-dimethoxy) and an organic acid (quinic acid). Leaves of C. xanthocarpa have high concentration of antioxidant compounds and it is a promising plant for the development of applications in the food, cosmetic, and pharmaceutical fields. The extraction of antioxidant compounds can add value to the productive chain of this plant.

Las hojas de Campomanesia xanthocarpa son un subproducto de la producción de frutos sin estudios sobre la actividad antioxidante. Así, este estudio tuvo como objetivo identificar los compuestos antioxidantes de las hojas de C. xanthocarpa mediante cromatografía líquida de ultra alta resolución acoplada con espectrometría de ionización-cuadrupolo-tiempo de vuelo-masa por electropulverización (UHPLC-ESI / qTOF) y mediante diferentes métodos antioxidantes in vitro. El extracto crudo de hojas de C. xanthocarpa tuvo un rendimiento del 15,2% y solo cinco de las 37 fracciones del extracto crudo tuvieron actividad antioxidante. El extracto crudo presentó mayor actividad antioxidante que las fracciones aisladas. Los compuestos antioxidantes identificados fueron ácidos fenólicos (ácido gálico y ácido clorogénico), flavonoides (quercetina y naringenina 7,4'-dimetoxi) y un ácido orgánico (ácido quínico). Las hojas de C. xanthocarpa tienen una alta concentración de compuestos antioxidantes y es una planta prometedora para el desarrollo de aplicaciones en los campos alimentario, cosmético y farmacéutico. La extracción de compuestos antioxidantes puede agregar valor a la cadena productiva de esta planta.

Antimicrobial activity of Tetradenia riparialeaf essential oil / Actividad antimicrobiana del aceite esencial de hoja de Tetradenia riparia

Food spoilage is a widely neglected problem and the constant use of synthetic fungicides could develop resistant fungi. The objective of this study was to evaluate the chemical composition and antimicrobial activity of Tetradenia riparialeaf essential oil against foodborne disease microorganisms. Leaf essential oil was obtained by hydrodistillation and identified by gas chromatography coupled to mass spectrometry. The antimicrobial activity was studied by broth microdilution. The major compounds identified were oxygenated sesquiterpenes (43.6%): 14-hydroxy-9-epi-(E)-cariophylene (20.8%) and τ-cadinol (18.4%); followed by oxygenated diterpenes (24.6%): 6,7-dehydroroyleanone (12.6%) and 9ß, 13ß-epoxy-7-abiethene (10.6%); sesquiterpenic hydrocarbons (17.1%) and oxygenated monoterpenes (7.4%): fenchone (5.6%). The essential oil had broad antibacterial and antifungal activity, mainly against A. versicolor and P. ochrochloron with fungistatic and fungicidal activities and B. cereus, L. monocytogenes, and S. aureuswith bacteriostatic and bactericidal activities. T. riparialeaf essential oil is a potential alternative to control microorganisms-

El deterioro de los alimentos es un problema ampliamente desatendido y el uso constante de fungicidas sintéticos podría desarrollar hongos resistentes. El objetivo de este estudio fue evaluar la composición química y la actividad antimicrobiana del aceite esencial de hoja de Tetradenia riparia contra microorganismos patógenos transmitidos por los alimentos. El aceite esencial de hoja se obtuvo por hidrodestilación y se identificó mediante cromatografía de gases acoplada a espectrometría de masas. La actividad antimicrobiana estudiada fue por microdilución en caldo. Los compuestos principales del aceite esencial se identificaron como sesquiterpenos oxigenados (43,6%): 14-hidroxi-9-epi-(E)-cariofileno (20,8%) y τ-cadinol (18,4%); seguido de diterpenos oxigenados (24,6%): 6-7-deshidroroileanona (12,6%) y 9ß, 13ß-epoxi-7-abieteno (10,6%); hidrocarburos sesquiterpénicos (17,1%) y monoterpenos oxigenados (7,4%): fenchona (5,6%). Tenía amplia actividad antibacteriana y antifúngica, principalmente contra A. versicolor y P. ochrochloron con actividades fungistáticas y fungicidas, y principalmente contra B. cereus, L. monocytogenes y S. aureus con actividades bacteriostáticas y bactericidas. El aceite esencial de hoja de T. riparia es una alternativa potencial para controlar microorganismos.

Larvicidal activity of Brunfelsia uniflora extracts on Aedes aegypti larvae.

The aim of this study was to evaluate the chemical composition and the larvicidal activity of Brunfelsia uniflora leaf and flower extracts against Aedes aegypti larvae. Twenty-four compounds were found in the leaf extract, and the major compounds were phytol (23.1%), 9,12,15-octadecatrienoic acid, ethyl ester (21.3%), and hexadecanoic acid, ethyl ester (12.8%). In the flower extract, twenty-four compounds were also identified and the major compounds were α-amyrin (35.7%), ß-amyrin (16.4%), and (EE)-geranyl linalool (9.6%) by gas chromatography coupled to mass spectrometry. The larvicidal activity was evaluated by larval immersion test. The lethal concentrations (LC) obtained from leaf extract were LC50 = 4.89 and LC99.9 = 11.14 mg/mL and from flower extract were LC50 = 3.82 and LC99.9 = 11.03 mg/mL, and the positive control presented LC50 = 0.40 and LC99.9 = 1.14 mg/mL. Thus, B. uniflora extracts are promising alternatives to control A. aegypti larvae.

Chemical composition and insecticidal activity of Garcinia gardneriana (Planchon & Triana) Zappi (Clusiaceae) essential oil / Composición química y actividad insecticida del aceite esencial de Garcinia gardneriana (Planchon & Triana) Zappi (Clusiaceae)

The present study aimed to analyze the chemical composition of the essential oil from Garcinia gardneriana (Planchon & Triana) Zappi leaves and fruits, and to determine its acaricidal activity on Rhipicephalus microplusy larval packet test and larvicidal action on Aedes aegyptiby larval immersion test. The chemical analysis of the essential oil by gas chromatography-mass spectrometry identified sesquiterpene hydrocarbons and oxygenated sesquiterpenes in bacupari leaves and fruits, and α-cedrene, α-chamigrene, α-trans-bergamotene, and ß-curcumene as major compounds. Essential oil from leaves of G. gardneriana presented acaricidal activity on R. microplus (LC50= 4.8 mg/mL; LC99= 10.8 mg/mL) and larvicidal effect on A. aegypti (LC50= 5.4 mg/mL; LC99 = 11.6 mg/mL), where as essential oil from the fruits of G. gardneriana showed LC50= 4.6 mg/mL and LC99= 8.9 mg/mL against R. microplus and LC50= 6.4 mg/mL and LC99= 13.9 mg/mL against A. aegypti. These results thus demonstrate the potential acaricidal and larvicidal activity of essential oil of G. gardneriana, offering new perspectives for the realization of bioassays from this essential oil.

El presente estudio tuvo como objetivo analizar la composición química del aceite esencial de las hojas y frutos de Garcinia gardneriana (Planchon & Triana) Zappi, y determinar su actividad acaricida en Rhipicephalus microplus y larvicida en Aedes aegypti empleando la prueba de inmersión de larvas. El análisis químico del aceite esencial por cromatografía de gases-espectrometría de masas identificó hidrocarburos sesquiterpénicos y sesquiterpenos oxigenados en hojas y frutos de bacupari, y α-cedreno, α-chamigreno, α-trans-bergamoteno y ß-curcumeno como compuestos principales. El aceite esencial obtenido de las hojas de G. gardneriana presentó actividad acaricida en la garrapata del ganado (LC50= 4,8 mg/mL; LC99= 10,8 mg/mL) y actividad larvicida en A. aegypti (LC50= 5,4 mg/mL; LC99= 11,6 mg/mL), así como, el aceite esencial obtenido de los frutos de G. gardneriana mostró LC50= 4,6 mg/mL y LC99= 8,9 mg/mL contra las larvas de garrapatas de ganado y LC50= 6,4 mg/mL y LC99= 13,9 mg/mL en las larvas de A. aegypti. Por lo tanto, estos resultados demuestran la actividad acaricida y larvicida del aceite essencial de G. gardneriana, ofreciendo nuevas perspectivas para la realización de bioensayos a partir de este aceite esencial.

Screening of six medicinal plant species for antileishmanial activity.

This study is aimed to investigate the in vitro anti-leishmanial activity of ethanolic, aqueous or dichloromethane extracts of leaves, flowers, fruits or roots, of six medicinal plant species, namely, Nectandra megapotamica, Brunfelsia uniflora, Myrcianthes pungens, Anona muricata, Hymenaea stigonocarpa and Piper corcovandesis. After isolation and analysis of chemical components by ultra-high performance liquid chromatography-high-resolution tandem mass spectrometry (UHPLC-HRMS/MS), the extracts were also tested for toxicity in J774.A1 macrophages and human erythrocytes. Phenolic acids, flavonoids, acetogenins, alkaloids and lignans were identified in these extracts. Grow inhibition of promastigotes forms of Leishmania amazonensis and Leishmania braziliensis and the cytotoxicity in J774.A1 macrophages were estimated by the XTT method. The most promising results for L. amazonensis and L. braziliensis were shown by the ethanolic extract of the fruits of Hymenaea stigonocarpa and dichloromethane extract of the roots of Piper corcovadensis, with IC 50 of 160 and 150 µg mL-1, resp. Ethanolic extracts of A. muricata (leaf), B. uniflora (flower and leaf), M. pungens (fruit and leaf), N. megapotamica (leaf), and aqueous extract of H. stigonocarpa (fruit) showed IC 50 > 170 µg mL-1 for L. amazonensis and > 200 µg mL-1 for L. braziliensis. The extracts exhibited low cytotoxicity towards J774.A1 macrophages with CC 50 > 1000 µg mL-1 and hemolytic activity from 0 to 46.1 %.

Crude extract of the tropical tree Gallesia integrifolia (Phytolaccaceae) for the control of Aedes aegypti (Diptera: Culicidae) larvae / Extracto crudo del árbol tropical Gallesia integrifolia (Phytolaccaceae) para el control de larvas de Aedes aegypti (Diptera: Culicidae)

Introduction: Phytoinsecticides are alternatives to control insects in different stages, Gallesia integrifolia (Spreng.) Harms, Phytolacaceae family, popularly known as pau d'alho, garlic tree, and guararema in Brazil, is known due to its strong alliaceous odor because of the presence of sulfur molecules in the plant. This species presents biological activity and potential insecticide effect that is still unexploited. Objective: This study aimed to evaluate the biological activity of the ethanolic crude extract from G. integrifolia leaves, flowers, and fruits on the control of Aedes aegypti third-stage larvae and pupae. Methods: The botanical material was collected in city Umuarama, Paraná, Brazil at the coordinates (23º46'16" S & 53º19'38" WO), and altitude of 442 m, the fruits of G. integrifolia were collected in May and the leaves and flowers in December 2017. The crude extracts of G. integrifolia leaves, flowers, and fruits were prepared by dynamic maceration technique. The chemical composition of the extracts was determined by gas chromatography coupled to a mass spectrometry. The insecticidal activity of the crude extracts of G. integrifolia were carried out on larvae and pupae of A. aegypti in concentrations between 0.001 to 25 000 mg/mL, and afterwards the lethal concentrations that kill 50 % (LC50) and 99.9 % (LC99.9) were determined by probit analysis. Anticholinesterase activity was determined by bioautographic method at concentrations from 0.000095 to 50 mg/mL. Results: The yield of G. integrifolia crude extracts were 8.2, 9.1, and 17.3 % for flowers, fruits, and leaves, respectively. The chemical composition of G. integrifolia extracts was characterized by presence of fatty acid esters, phytosterols, vitamins, oxygenated diterpenes and organosulfur compounds. The flower extract presented the high amount of sulfur compounds (20.2 %) such as disulfide, bis (2-sulfhydryl ethyl) (11.9 %), 2,3,5-trithiahexane (6.2 %), 1,2,4-trithiolane (1.1 %), and 2,4-dithiapentane (1.1 %). Regarding the insecticidal activity, flower extract showed highly active with LC99.9 of 0.032 mg/mL and LC99.9 of 0.969 mg/mL on A. aegypti larvae and pupae, respectively, and the highest inhibition of acetylcholinesterase enzyme (0.00019 mg/mL) ex situ. The flower extract presented anticholinesterase and larvicide activity, respectively, 12.8 % and 35.6 % greater than the control temephos. Conclusions: This study opens new perspectives on the use of extracts from G. integrifolia as a bioinsecticide alternative for the control of A. aegypti larvae and pupae.

Introducción: Los fitoinsecticidas son alternativas para el control de insectos en diferentes etapas, Gallesia integrifolia (Spreng.) Harms, familia Phytolacaceae, conocida popularmente como pau d'alho, árbol de ajo y guararema en Brasil, es conocida por su fuerte olor aliáceo debido a la presencia de moléculas de azufre en la planta. Esta especie presenta actividad biológica y potencial efecto insecticida que aún no está explotado. Objetivo: El objetivo de este estudio fue evaluar la actividad biológica del extracto crudo etanólico de las hojas, flores y frutos de G. integrifolia en el control de las larvas y pupas de la tercera etapa de Aedes aegypti. Métodos: El material botánico fue recolectado en la ciudad de Umuarama, Paraná, Brasil (23º46'16" S & 53º19'38" W), a una altitud de 442 m y los frutos de G. integrifolia fueron recolectados en mayo de 2017 y las hojas y flores en diciembre del mismo año. Los extractos crudos de hojas, flores y frutos de G. integrifolia se prepararon por técnica de maceración dinámica. La composición química de los extractos se determinó por cromatografía de gases acoplada a espectrometría de masas. La actividad insecticida de los extractos crudos de G. integrifolia fue evaluada en larvas y pupas de A. aegypti en concentraciones entre 0.001 a 25 000 mg/mL, y las concentraciones letales que matan 50 % (CL50) y 99.9 % (CL99.9) de larvas fueron determinados por análisis de probit. La actividad anticolinesterasa fue evaluada por método bioautográfico en concentraciones de 0.000095 a 50 mg/mL. Resultados: El rendimiento de los extractos crudos de G. integrifolia fue de 8.2, 9.1 y 17.3 % para flores, frutos y hojas, respectivamente. La composición química de los extractos de G. integrifolia se caracterizó por la presencia de ésteres de ácidos grasos, fitosteroles, vitaminas, diterpenos oxigenados y compuestos organosulfurados. El extracto de las flores presentó alta cantidad de compuestos de azufre (20.2 %) como disulfuro, bis(2-sulfhidril etilo) (11.9 %), 2,3,5-tritiahexano (6.2 %), 1,2,4-tritiolano (1.1 %) y 2,4-ditiapentano (1.1 %). En relación con la actividad insecticida, el extracto de las flores mostró una gran actividad con CL99.9 de 0.032 mg/mL y CL99.9 de 0.969 mg/mL en larvas y pupas de A. aegypti, respectivamente, y la inhibición más alta de la enzima acetilcolinesterasa (0.00019 mg/mL) ex situ. El extracto de las flores presentó actividad anticolinesterasa y larvicida, 12.8 y 35.6 %, respectivamente, mayor que el control temephos. Conclusiones: Este estudio abre nuevas perspectivas sobre el uso de extractos de G. integrifolia como alternativa bioinsecticida para el control de larvas y pupas de A. aegypti.

Larvicidal activity of piperovatine and dichloromethane extract from Piper corcovadensis roots against mosquitoes Aedes aegypti L / Actividad larvicida de piperovatina y extracto de diclorometano de las raíces de Piper corcovadensis contra mosquitos vectores del dengue Aedes aegypti L

The research of new substances capable of controlling the Aedes aegypti mosquito is urgent due to the increase in the transmission of the diseases such as dengue, chikungunya and Zika virus by the vector. Thus, the aim of this study was to evaluate the larvicidal activity of crude extract of Piper corcovadensis roots, a native plant from Brazil, and of the isolated compound piperovatine against larvae of A. aegypti by the larval immersion test. The lethal concentration that killed 50% (LC50) and 99% (LC99) of larvae was determined by Probit analysis. The results indicated high larvicidal activity on A. aegypti larvae for crude extract of Piper corcovadensis roots with LC50 of 4.86 µg/mL and LC99 of 15.50 µg/mL and piperovatine with LC50 of 17.78 µg/mL and LC99 of 48.55 µg/mL. This work opens new perspectives to the development of future products with crude extract of Piper corcovadensis roots and piperovatine that can be applied to mosquito control.

La investigación de nuevas sustancias capaces de controlar el mosquito Aedes aegypti es urgente debido al aumento en la transmisión de enfermedades como el dengue, el chikungunya y el virus Zika por el vector. Por lo tanto, el objetivo de este estudio fue evaluar la actividad larvicida del extracto crudo de las raíces de Piper corcovadensis, una planta nativa de Brasil, y del compuesto aislado piperovatine contra larvas de A. aegypti mediante la prueba de inmersión larvaria. La concentración letal que mató al 50% (LC50) y al 99% (LC99) de larvas se determinó mediante análisis Probit. Los resultados indicaron una alta actividad larvicida en larvas de A. aegypti para extracto crudo de las raíces de Piper corcovadensis con LC50 de 4.86 µg/mL y LC99 de 15.50 µg/mL y piperovatine con LC50 de 17.78 µg/mL y LC99 de 48.55 µg/mL. Este trabajo abre nuevas perspectivas para el desarrollo de futuros productos con extracto crudo de las raíces de Piper corcovadensis y piperovatine que pueden aplicarse al control de mosquitos.

Essential oil and fractions isolated of Laurel to control adults and larvae of cattle ticks.

This study, was to evaluate the acaricidal effect of the essential oil (EO) and fractions (FR) obtained from Laurus nobilis leaves on Rhipicephalus (Boophilus) microplus. Eight fractions were obtained, however FR1: sabinene (37.83%), ß-pinene (13.50%), 1,8-cineole (12.66%), α-pinene (12.56%) and FR8: α-terpineol (79.19%) were highlighted as to the larvicidal potential when submitted by Larval Packet Test. The EO was tested by the Adult Immersion Test, at concentrations of 200.00; 100.00 and 50.00 µL/mL caused mortality of engorged females, egg mass reduction and hatching inhibition. Two fractions are shown to be efficient in controlling larvae FR8 (LC50 = 0.13 µL/mL, LC99 = 0.51 µL/mL) and FR1 (LC50 = 0.20 µL/mL, LC99 = 0.56 µL/mL). The fractionation of EO was determinant to elucidate which compounds were responsible for the larvicidal potential. This study opens new perspectives to direct new bioassays with the compounds obtained in the fractionation, since they present high potential on cattle tick larvae.

Antioxidant activity of α and ß-amyrin isolated from Myrcianthes pungens leaves.

Brazil has a great variety of native plants which could be explored and among them guabiju (Myrcianthes pungens) stands out. Thus, this study consisted of isolating pentacyclic triterpenes, α and ß-amyrin from guajibu leaves, and determine the antioxidant activity.The leaves were dried, pulverized and submitted to a dynamic maceration process with ethanol 96° GL, concentrated to obtain crude leaf extract (CLE). CLE was eluted with dichloromethane until total depletion, resulting in a dichloromethane fraction (FRDicl) which was concentrated and analyzed by GC/MS and NMR. The result of the chemical analysis revealed the presence of pentacyclic triterpenes of oleanane (ß-amyrin), and ursane (α-amyrin) groups. The ß-carotene bleaching method revealed a high antioxidant activity for the CLE as well as for FRDicl. The antioxidant protection equivalent to the trolox was of 137 and 129%, respectively at 500 µg/mL. The antioxidant potential of FRDicl can be explained by the presence of α and ß-amyrins.

Ginger essential oil and fractions against Mycobacterium spp.

ETHNOPHARMACOLOGICAL RELEVANCE: Zingiber officinale (ginger) is a perennial herbaceous plant native in tropical Asia and generally cultivated in most American tropical countries with widespread use in popular medicine. Ginger essential oil (GEO) has been reported to exhibit several biological activities, such as antimicrobial. AIMS OF THE STUDY: The aim of this study was to determine the composition and the property of GEO and related fractions against Mtb and NTM, as well as their cytotoxicity. METHODS AND MATERIALS: GEO was obtained by hydrodistillation and fractionation was performed. Chemical characterization of GEO and fractions were carried out by gas chromatography/mass spectrometry. The antimycobacterial activity was evaluated by resazurin microtiter assay plate and broth microdilution method for Mtb and NTM, respectively. The cytotoxicity in Vero cells was assessed by MTT colorimetric assay. RESULTS: The analyses showed 63 compounds in the GEO sample, characterized by a high number of monoterpenes and sesquiterpenes. GEO fractionation rendered 11 fractions (FR1 to FR11). GEO and fractions minimum inhibitory concentration ranged from 31.25 to >250 µg/mL against Mtb and from 15.6 to >250 µg/mL against NTM. GEO showed better activity against NTM, M. chelonae, and M. abscessus sub. massiliense, than the semi-pure fractions. One fraction (FR5), containing γ-eudesmol as the main compound, was the most active against Mtb and NTM. The GEO and semi-pure fractions cytotoxicity assay showed CC50 63.3 µg/mL, and 36.3-312.5 µg/mL, respectively. CONCLUSIONS: In general, GEO showed a mix of monoterpenes and sesquiterpenes and a better antimycobacterial activity than the semi-pure fractions. Cytotoxic effects of GEO and its fractions should be better investigated.

In vitro antioxidant potential and in vivo effects of Schinus terebinthifolia Raddi leaf extract in diabetic rats and determination of chemical composition by HPLC-ESI-MS/MS.

The present study investigated the in vitro and in vivo antioxidant potential and phytochemical composition of Schinus terebinthifolia, which is widely used in folk medicine for various therapeutic purposes. The in vitro analyses indicated that the hydroethanolic extract (HE) had 312.50 ± 0.50 mg GAE/g of total phenols. It also presented anti-DPPH• and anti-ABTS•+ activity, reduced phosphomolybden and metal ions and blocked the bleaching of ß-carotene. The HE at concentrations of 3.0 and 2.0 µg/mL had TRAP values of 2.223 ± 0.018 and 1.894 ± 0.026 µM Trolox, respectively. The HE increased the availability of antioxidants in plasma in treated animals in vivo. HPLC-ESI-MS/MS indicated the presence of 11 phenols: cumaric acid, (+)-catechin, myricetin-3-O-glicuronide, kaempferol-3-O-glucoside, myricetin, myricitrin, quercetin, gallic acid, methyl galate, pentagalloyl glucose and ethyl galate. Thus, S. terebinthifolia has potential for the prevention or treatment of diseases that are related to oxidative stress, such as diabetes mellitus.

Antimicrobial and Antioxidant Activities of the Extract and Fractions of Tetradenia riparia (Hochst.) Codd (Lamiaceae) Leaves from Brazil.

Tetradenia riparia (Lamiaceae) is native to Central Africa popularly known as myrrh, used in folk medicine to treat various diseases like malaria, gastroenteritis, and tropical skin disease. This research was to evaluate the antioxidant and antibacterial activities of the crude extract (CE) and fractions (FR) of the T. riparia by classical chromatography. The CE of T. riparia leaves was submitted to column chromatographic fractionation to obtain four fractions of the interest, which were identified by nuclear magnetic resonance and gas chromatograph coupled to mass spectrum: FR-I (abieta-7,9(11)-dien-13-ß-ol), FR-II (Ibozol), FR-III (8 (14), 15-sandaracopimaradiene-2α, 18-diol and 8 (14), 15-sandaracopimaradiene-7α, 18-diol), and FR-IV (Astragalin, Boronolide and Luteolin). Total phenol content of CE and FR were measured, and antioxidant action by methods of DPPH (2,2-diphenyl-1-picrylhydrazyl), ß-carotene/linoleic acid system, and ferric reducing/antioxidant power (FRAP) and the antibacterial activity was evaluated by the broth microdilution method with the determination of the minimum inhibitory concentration (MIC). The FR-IV presented antioxidant potential with 181.67 µg gallic acid/mg, IC50 of 0.61 µg/mL by DPPH method, 55.61% oxidation protection by ß-carotene/linoleic acid system and 4.59 µM ferrous sulfate/mg of sample by FRAP, and the FR-I showed higher antibacterial potential on the strain Staphylococcus aureus with MIC 0.98 µg/mL, Enterococcus faecalis and Bacillus cereus with MIC 31.2 µg/mL. Thus, the fractionation of CE was extremely important to detect fractions with potential activities, and investigations are necessary regarding the mechanism of action and action in vivo.

Ex vivo cultures of glioblastoma in three-dimensional hydrogel maintain the original tumor growth behavior and are suitable for preclinical drug and radiation sensitivity screening.

Identification of new drugs and predicting drug response are major challenges in oncology, especially for brain tumors, because total surgical resection is difficult and radiation therapy or chemotherapy is often ineffective. With the aim of developing a culture system close to in vivo conditions for testing new drugs, we characterized an ex vivo three-dimensional culture system based on a hyaluronic acid-rich hydrogel and compared it with classical two-dimensional culture conditions. U87-MG glioblastoma cells and seven primary cell cultures of human glioblastomas were subjected to radiation therapy and chemotherapy drugs. It appears that 3D hydrogel preserves the original cancer growth behavior and enables assessment of the sensitivity of malignant gliomas to radiation and drugs with regard to inter-tumoral heterogeneity of therapeutic response. It could be used for preclinical assessment of new therapies.

Cerebral biochemical pathways in experimental autoimmune encephalomyelitis and adjuvant arthritis: a comparative metabolomic study.

Many diseases, including brain disorders, are associated with perturbations of tissue metabolism. However, an often overlooked issue is the impact that inflammations outside the brain may have on brain metabolism. Our main goal was to study similarities and differences between brain metabolite profiles of animals suffering from experimental autoimmune encephalomyelitis (EAE) and adjuvant arthritis (AA) in Lewis rat models. Our principal objective was the determination of molecular protagonists involved in the metabolism underlying these diseases. EAE was induced by intraplantar injection of complete Freund's adjuvant (CFA) and spinal-cord homogenate (SC-H), whereas AA was induced by CFA only. Naive rats served as controls (n = 9 for each group). Two weeks after inoculation, animals were sacrificed, and brains were removed and processed for metabolomic analysis by NMR spectroscopy or for immunohistochemistry. Interestingly, both inflammatory diseases caused similar, though not identical, changes in metabolites involved in regulation of brain cell size and membrane production: among the osmolytes, taurine and the neuronal marker, N-acetylaspartate, were decreased, and the astrocyte marker, myo-inositol, slightly increased in both inoculated groups compared with controls. Also ethanolamine-containing phospholipids, sources of inflammatory agents, and several glycolytic metabolites were increased in both inoculated groups. By contrast, the amino acids, aspartate and isoleucine, were less concentrated in CFA/SC-H and control vs. CFA rats. Our results suggest that inflammatory brain metabolite profiles may indicate the existence of either cerebral (EAE) or extra-cerebral (AA) inflammation. These inflammatory processes may act through distinct pathways that converge toward similar brain metabolic profiles. Our findings open new avenues for future studies aimed at demonstrating whether brain metabolic effects provoked by AA are pain/stress-mediated and/or due to the presence of systemic proinflammatory molecules. Regardless of the nature of these mechanisms, our findings may be of interest for future clinical studies, e.g. by in-vivo magnetic resonance spectroscopy.

Pilocytic astrocytoma of the optic pathway: a tumour deriving from radial glia cells with a specific gene signature.

Pilocytic astrocytomas are WHO grade I gliomas that occur predominantly in childhood. They share features of both astroglial and oligodendroglial lineages. These tumours affect preferentially the cerebellum (benign clinical course) and the optic pathway, especially the hypothalamo-chiasmatic region (poor prognosis). Understanding the molecular basis responsible for the aggressive behaviour of hypothalamo-chiasmatic pilocytic astrocytomas is a prerequisite to setting up new molecular targeted therapies. We used the microarray technique to compare the transcriptional profiles of five hypothalamo-chiasmatic and six cerebellar pilocytic astrocytomas. Validation of the microarray results and comparison of the tumours with normal developing tissue was done by quantitative real-time PCR and immunohistochemistry. Results demonstrate that cerebellar and hypothalamo-chiasmatic pilocytic astrocytomas are two genetically distinct and topography-dependent entities. Numerous genes upregulated in hypothalamo-chiasmatic pilocytic astrocytomas also increased in the developing chiasm, suggesting that developmental genes mirror the cell of origin whereas migrative, adhesive and proliferative genes reflect infiltrative properties of these tumours. Of particular interest, NOTCH2, a gene expressed in radial glia and involved in gliomagenesis, was upregulated in hypothalamo-chiasmatic pilocytic astrocytomas. In order to find progenitor cells that could give rise to hypothalamo-chiasmatic pilocytic astrocytomas, we performed a morphological study of the hypothalamo-chiasmatic region and identified, in the floor of the third ventricle, a unique population of vimentin- and glial fibrillary acidic protein-positive cells highly suggestive of radial glia cells. Therefore, pilocytic astrocytomas of the hypothalamo-chiasmatic region should be considered as a distinct entity which probably originates from a unique population of cells with radial glia phenotype.

Thalamic gliomas in children: an extensive clinical, neuroradiological and pathological study of 14 cases.

OBJECT: Thalamic tumors represent only 1 to 5% of brain neoplasms but frequently affect children. However, pediatric series are rare and go back to several years in spite of recent advances in the neuroradiological, pathological, and molecular fields. METHODS: We report a series of 14 pediatric thalamic gliomas with clinical, neuroradiological, and pathological studies including p53 immunostaining in 11 cases and 1p19q status in three cases. RESULTS: Our series included five pilocytic astrocytomas, seven oligodendrogliomas, and two glioblastomas. Pilocytic astrocytomas were characterized by strong contrast enhancement, lack of p53 expression, and excellent prognosis. Oligodendrogliomas frequently demonstrated an aspect of unilateral thalamic enlargement lacking or with slight contrast enhancement. Some of them expressed p53 or demonstrated 1p loss. Anaplastic oligodendrogliomas and glioblastomas displayed a poor outcome, with a mean survival of 8 months after surgery. CONCLUSION: Our series of pediatric thalamic gliomas clearly distinguishes pilocytic astrocytomas from anaplastic oligodendrogliomas regarding neuroimaging, pathology, and prognosis.