Actividad antiparasitaria in vitro de extractos etanólicos de Erythrina crista-galii L., Erythrina falcata Benth y Erythrina dominguezii Hassl / In vitro antiparasitic activity of ethanolic extracts of Erythrina crista-galii L., Erythrina falcata Benth and Erythrina dominguezii Hassl

La tripanosomiasis americana y la leishmaniosis son problemas de salud pública relevantes en Iberoamérica. Se evaluó la actividad antiparasitaria in vitro de extractos de cortezas y hojas de Erythrina crista-galli L., E. dominguezii Hassl y E. falcata Benth sobre formas tripomastigotas de Trypanosoma cruzi y promastigotas de Leishmania amazonensis, L. infantum L. braziliensis. El extracto de corteza de E. dominguezii Hassl presentó CI50 de 11,7 µg/ml y 14,9 µg/ml con dos cepas de T. cruzi, Ypsilon y Cl Brener. El extracto de hoja presentó CI50 mayor a 250 µg/ml. La corteza de E. falcata Benth tuvo CI50 de 11,29 µg/ml y 7,37 µg/ml con Ypsilon y Cl Brener y el extracto de hojas CI50 de 37,2 µg/ml y 25,68 µg/ml. El extracto de hojas de E. crista-galli L. presentó CI50 mayor a 250 µg/ml, y el de corteza CI50 11,2µg/ml y 7,1µg/ml con Ypsilon y Cl Brener. El extracto de corteza de E. dominguezii Hassl presentó CI50 de 3,22 µg/ml, 6,94 µg/ml y 1,9 µg/ml para L. amazonensis, L. infantum y L. braziliensis y en hojas la CI50 fue mayor a 100 µg/ml. La corteza de E. crista-galli L. tuvo CI50 de 2,1 µg/ml, 2,09 µg/ml y 1,6 µg/ml para las tres especies. El extracto de hoja tuvo una CI50 mayor a 100 µg/ml. El extracto de corteza de E. falcata Benth tuvo CI50 de 2,17 µg/ml, 2,09 µg/ml y 1,52 µg/ml con las tres especies de Leishmania y el de hoja CI50 de 14,9 µg/ml, 10,4 µg/ml y 11,6 µg/ml. Los extractos de corteza podrían ser una alternativa para el tratamiento de estas enfermedades parasitarias, previa confirmación con estudios in vivo y toxicidad

Harvesting canthinones: identification of the optimal seasonal point of harvest of Zanthoxylum chiloperone leaves as a source of 5-methoxycanthin-6-one.

This article is focused on the seasonal variation in the contents of 5-methoxycanthin-6-one from the leaves of Zanthoxylum chiloperone (Rutaceae). Based on the pharmacological interest presented by 5-methoxycanthin-6-one, its seasonal variation in Z. chiloperone leaves was analysed in order to determine the best time for harvesting, optimising the 5-methoxycanthin-6-one content. The seasonal dynamics of canthinone alkaloids can be the key to improve the isolation from natural sustainable sources, such as leaves. Complementarily, this study describes the phytochemistry of leaf from this Ruraceae species.

Antiproliferative activity of trans-avicennol from Zanthoxylum chiloperone var. angustifolium against human cancer stem cells.

Zanthoxylum chiloperone var. angustifolium root bark was studied with the aim of finding novel molecules able to overcome cancer stem cell chemoresistance. Purification of a methanol-soluble extract resulted in the isolation of a known pyranocoumarin, trans-avicennol (1). Compound 1 demonstrated antiproliferative activity on glioma-initiating cells, whereas it was inactive on human neural stem cells. trans-Avicennol (1) activated the MAPK/ERK pathway and was also evaluated for its ability to inhibit the enzyme indoleamine-2,3-dioxygenase.

The antiplasmodium effects of a traditional South American remedy: Zanthoxylum chiloperone var. angustifolium against chloroquine resistant and chloroquine sensitive strains of Plasmodium falciparum

Zanthoxylum chiloperone var. angustifolium Engl., Rutaceae, is used in traditional medicine to treat fungal and protozoal infections in the central area of South America. Considering the increasing resistance of Plasmodium falciparum in malarial ridden areas, we explored the anti-plasmodial effects of three compounds isolated from Z. chiloperone. The pyranocoumarin transavicennol and the canthinone alkaloids, canthin-6-one and 5-methoxycanthin-6-one, were found to have IC50 on chloroquine/mefloquine resistant and sensitive strains of P. falciparum of 0.5-2.7, 2.0-5.3 and 5.1-10.4 ƒÊg/mL, respectively. Moreover, the formation of heme adducts by these compounds is described by a novel alternative method based on MS-CID methods. The alkylamide sanshool was also identified, for first time in this plant, in the dichloromethanic and ethanolic extracts and the extracts were found to be notably non-toxic and displayed good anti-plasmodial effects.

Zanthoxylum chiloperone leaves extract: first sustainable Chagas disease treatment.

ETHNOPHARMACOLOGICAL RELEVANCE: Zanthoxylum chiloperone var. angustifolium Engl. (Rutaceae) stem bark is used traditionally in Paraguay for its antiparasitic properties. Canthin-6-one is main compound isolated from Zanthoxylum chiloperone var angustifolium with broad spectrum antifungal, leishmanicidal and trypanocidal activities. AIM OF THE STUDY: The qualitative and quantitative characterization and the isolation of main alkaloidal components of different organs of Zanthoxylum chiloperone are investigated by HPLC-UV-MS. The in vitro biological activity of each extract against trypomastigote and amastigote forms of Trypanosoma cruzi parasites were evaluated, then comparison the in vivo efficacy of the ethanolic leaves extract of Zanthoxylum chiloperone with reference drug, benznidazole, in acute Trypanosoma cruzi infected mice when administered by oral route. We have also evaluated the mutagenic and cytotoxic activity of the main component of Zanthoxylum chiloperone, i.e. canthin-6-one, by mouse bone marrow micronucleus test. MATERIALS AND METHODS: The compositions of the ethanol extracts obtained after the maceration process were studied by HPLC-UV-MS methods. The quantitation analysis was performed by external standard method, using a calibration curve constructed utilizing solutions containing different concentrations of the reference samples. The anti-trypomastigote activity was evaluated by the lysis effect on mouse blood trypomastigotes (Y strain Trypanosoma cruzi). The anti-amastigote Trypanosoma cruzi activity was evaluated by a modified colorimetric method with chlorophenol red-ß-d-galactopyranoside (CPRG). The cytotoxicity of extracts and compounds was performed on NCTC 929 cells. The in vivo efficacy of the ethanolic leaves extract of Zanthoxylum chiloperone and benznidazole, in acute Trypanosoma cruzi (two different strains) was evaluated in Trypanosoma cruzi infected mice; the drugs were administered by oral route. The mortality rates were recorded and parasitaemias in control and treated mice were determined once weekly for 70 days. The mutagenic and cytotoxic activity of the main component of Zanthoxylum chiloperone, canthin-6-one, by mouse bone marrow micronucleus test. RESULTS: Canthin-6-one was the main compound of stem and root bark and 5-methoxy-canthin-6-one in leaves and fruits. The ethanolic leaves extract, canthin-6-one and benznidazole presented, approximately, the same level of in vitro activity against trypomastigote and amastigote forms of Trypanosoma cruzi. We have also evaluated the mutagenic and cytotoxic effects of canthin-6-one by micronucleus test in mice. This test showed any mutagenic and cytotoxic damages. The effects of oral or subcutaneous treatments at 10 mg/kg daily for 2 weeks with the ethanolic extract of leaves of Zanthoxylum chiloperone were examined in Balb/c mice infected acutely with Trypanosoma cruzi (CL or Y strain) and compared with benznidazole at 50 mg/kg for 2 weeks. In these experiments, 70 days after infection, parasitaemia and serological response were significantly reduced with the oral ethanolic extract treatment compared with reference drug. CONCLUSIONS: This study have shown the efficacy of the leaves extract of Zanthoxylum chiloperone in reducing Trypanosoma cruzi parasitaemia in vivo assays and could be welcomed by scientific and rural communities of Paraguay because it could help them towards the use of local resources to treat an endemic infection, Chagas disease, affecting 20% of the population of this country.

Antileishmanial activity of furoquinolines and coumarins from Helietta apiculata.

UNLABELLED: The bark infusion of H. apiculata are used to treat wound healing related to cutaneous leishmaniasis and as anti-inflammatory. AIM OF THE STUDY: To isolate, purify active constituents of H. apiculata stem bark, and evaluate their in vitro and in vivo antileishmanial activities. MATERIALS AND METHODS: Isolation by chromatographic methods and chemical identification of furoquinoline alkaloids and coumarins, then evaluation of the in vitro leishmanicidal activity of these compounds against three strains of Leishmania sp. promastigotes and in vivo against Leishmania amazonensis in Balb/c mice. RESULTS: Furoquinoline alkaloids and coumarins presented a moderate in vitro activity against promastigote forms of Leishmania sp. with IC(50) values in the range between 17 and >50 microg/ml. Balb/c mice infected with Leishmania amazonensis were treated with gamma-fagarine by oral route, or with 3-(1'-dimethylallyl)-decursinol or (-)-heliettin by subscutaneous route for 14 days at 10mg/kg daily. In these conditions, gamma-fagarine, 3-(1'-dimethylallyl)-decursinol and (-)-heliettin showed the same efficacy as the reference drug reducing by 97.4, 95.6 and 98.6% the parasite loads in the lesion, respectively. CONCLUSION: These compounds showed significant efficacy in L. amazonensis infected mice, providing important knowledge to improve its potential role for a future use in the treatment of cutaneous leishmaniasis.

Actividad tripanocida in vitro e in vivo de extractos etanólicos de algunas plantas medicinales bolivianas / In vitro and in vivo trypanocidal activity, of ethanolic extracts from some bolivian medicinal plants

A 100 años del descubrimiento de la enfermedad de Chagas, aun no existen drogas que satisfagan completamente. Los extractos de plantas medicinales son una posibilidad de obtener nuevos compuestos que sean activos contra Trypanosoma cruzi. Se evaluó la actividad tripanocida in vitro sobre las formas tripomastigotes de ocho extractos etanólicos provenientes de 4 plantas medicinales bolivianas. El extracto etanólico de la corteza de Anacardium occidentale fue el más activo in vitro (CI50= 200 µg/ml), seguido de las hojas de Bowdichia virgilioides (350 µg/ml). A. occidentale, B. virgilioides y Senna reticulata, no son eficaces en la fase aguda de la enfermedad de Chagas en el modelo murino. (AU)

Activity of a hydroxybibenzyl bryophyte constituent against Leishmania spp. and Trypanosoma cruzi: in silico, in vitro and in vivo activity studies.

The synthesis and potent antiprotozoal activity of 14-hydroxylunularin, a natural hydroxybibenzyl bryophyte constituent is reported. 14-hydroxylunularin was highly active in vitro assays against culture and intracellular forms of Leishmania spp. and Trypanosoma. cruzi, in absence of cytotoxicity against mammalian cells. Preliminary structure-activity relationship studies showed that the reported bioactivity depends on hybridization at the carbon-carbon bridge, position and number of free hydroxy group on the aromatic rings. The reported results were also in agreement with the in silico prediction using Non-Stochastic Quadratic Fingerprints-based algorithms. The same compound also showed antiprotozoal activity in Leishmania spp. infected mice by oral and subcutaneous administration routes, with an optimal treatment of a daily subcutaneous administration of 10 mg/kg of body weight for 15 days. This study suggested that 14-hydroxylunularin may be chosen as a new candidate in the development of leishmanicidal therapy.

Phytochemical and antiprotozoal activity of Ocotea lancifolia.

Thirteen known isoquinoline alkaloids were isolated from Ocotea lancifolia, popularly known as << canela pilosa >> in Brasil and << laurel né >> by the Guarani people which means smell laurel. Their activities against the promastigote forms of three Leishmania strains and the bloodstream form of Trypanosoma cruzi were evaluated, as well as their hepatocytotoxicity. Among them, the noraporphine alkaloid (-) caaverine has shown the most interesting antiprotozoal activity against Leishmania and T. cruzi parasites.

Effects of canthin-6-one alkaloids from Zanthoxylum chiloperone on Trypanosoma cruzi-infected mice.

Canthin-6-one (1), isolated from Zanthoxylum chiloperone (Rutaceae), possesses a broad sprectum of antifungal and leishmanicidal activities. In this study, we have examined the antiparasitic effects of canthin-6-one (1), 5-methoxycanthin-6-one (2), canthin-6-one N-oxide (3), as well as that of the total alkaloids of Zanthoxylum chiloperone stem bark, in Balb/c mice infected either acutely or chronically with Trypanosoma cruzi. The compounds were administered orally or subcutaneously at 5mg/kg/day for 2 weeks, whereas the alkaloidal extract was given at 50mg/kg/day for 2 weeks. The antiparasitic activity was compared with that of benznidazole given at 50mg/kg/day for 2 weeks. In the case of acute infection, parasiteamia was significantly reduced following oral treatment with canthin-6-one (1). Moreover, the total alkaloids of Zanthoxylum chiloperone stem bark led to high levels of parasitological clearance. Seventy days post-infection, the serological response in the acute model was significantly different between oral canthin-6-one (1) and benznidazole-treated mice. Chronic model of the disease showed that both canthin-6-one (1) and the alkaloidal extract at the above dosage induced 80-100% animal survival compared to untreated controls. These results indicate that canthin-6-one (1) exhibits trypanocidal activity in vivo in the mouse model of acute or chronic infection. This is the first demonstration of anti-Trypanosoma cruzi activity for a member of this chemical group (canthinones). Considering the very low toxicity of canthin-6-one (1), our results suggest that long-term oral treatment with this natural product could prove advantageous compared to the current chemotherapy of Chagas disease.

Antifungal compounds from Zanthoxylum chiloperone var. angustifolium.

An alkaloidal extract of the stem barks of Zanthoxylum chiloperone var. angustifolium exhibited antifungal activity against Candida albicans, Aspergillus fumigatus and Trichophyton mentagrophytes var. interdigitale using a TLC bioautographic method. Bioassay-guided fractionation of this extract resulted in the isolation of two active compounds identi fi ed as canthin-6-one and 5-methoxycanthin-6-one. Canthin-6-one exhibited a broad spectrum of activities against Aspergillus fumigatus, A. niger, A. terreus, Candida albicans, C. tropicalis, C. glabrata, Cryptococcus neoformans, Geotrichum candidum, Saccharomyces cerevisiae, Trichosporon beigelii, Trichosporon cutaneum and Trichophyton mentagrophytes var. interdigitale with MICs values between 5.3 and 46 micro mol/L. 5-methoxy-canthin-6-one was active against only Trichophyton mentagrophytes var. interdigitale with a MIC value of 12.3 micro mol/L.

Evaluación de la técnica de hemocultivo y epidemiología de la enfermedad de Chagas en pacientes agudos y crónicos / Annual reports 1998 / Evaluation of the hemoculture technique and epidemiological characteristic of acute and chronic Chagas disease patients

El diagnóstico de la enfermedad de Chagas por métodos parasitológicos se realiza mediante la visualización directa o indirecta del parásito. La detección del mismo es relativamente fácil en la fase aguda, cuando la parasistemia es alta; sin embargo, durante las fases intermedias y la cronica se debe recurrir a métodos que requieren una observación más prologada como el xenodiagnóstico y el hemocultivo, siendo la sensibilidad de estos últimos menor. En este trabajo se determino la sensibilidad del hemocultivo en paciente chagasicos y se provee informaciones complementaria sobre las carácteristicas epidemiólogicas de los mismos. Se realizó un levantamiento retrospectivos de datos obtenidos de fichas de 100 pacientes chagasicos que consultaron en el Departamento de Medicina Tropical del IICS durante el periodo de enero 1984 a enero 1992. A todos estos pacientes se le realizo la prueba de hemocultivo en forma de rutina. El 21 por ciento de estos pacientes presentaron resultados positivos para T.cruzi por la tecnica de hemocultivo. Teniendo en cuenta el cuadro clinico de los pacientes, se observo que el 100 por ciento (4/4) de los casos agudos, el 73,2 por ciento (6/25) de los pacientes con lesiones cardiacas o digestivas y el 16,4 por ciento (11/67) de los individuos en la etapa indeterminada fueron positivos a la T.cruzi y el resto de los pacientes sin diagnostico clinico fue negativo por esta técnica. Teniendo en cuenta la edad de estos pacientes se encontro que los menores de 30 años dieron hemocultivo positivo más frecuentemente que aquellos que pertenecieron a grupos de mayor edad. En este estudio, la psibilidad del hemocultivo mostro cierta asociación con la edad y la procedencia del paciente chagasico

Producciòn local de Leishmanina / Local Leishmanin production / Annual reports 1996

La prueba cutànea de la leishmanina (reacciòn de Montenegro) es ùtil como mètodo de tamizaje para el diagnòstico epidemiològico de las leishmaniasis, permitiendo definir las zonas endèmicas para la enfermedad. Sòlo con una buena confiabilidad en los antìgenos se asegura que la prueba cutànea responda a las expectativas en estudios epidemiològicos. En el IICS se han producido hasta el momento seis lotes de dicho antìgeno a partir de promastigotes de L. amazonensis, que han aprobado los diversos ensayos de control, segùn dictados internacionales para productos de administraciòn parenteral. De todos los ensayos realizados, sòlo el ensayo de acividad, desarrollado a nivel experimental en animales, no arroja aùn resultados concluyentes. La disponibilidad de esta producciòn en el IICS permitirà desarrollar estudios epidemiològicos acerca de la prevalenciaq de la leishmaniasis en el Paraguay.

Producciòn local de Leishmanina / Local Leishmanin production / Annual reports 1996

La prueba cutànea de la leishmanina (reacciòn de Montenegro) es ùtil como mètodo de tamizaje para el diagnòstico epidemiològico de las leishmaniasis, permitiendo definir las zonas endèmicas para la enfermedad. Sòlo con una buena confiabilidad en los antìgenos se asegura que la prueba cutànea responda a las expectativas en estudios epidemiològicos. En el IICS se han producido hasta el momento seis lotes de dicho antìgeno a partir de promastigotes de L. amazonensis, que han aprobado los diversos ensayos de control, segùn dictados internacionales para productos de administraciòn parenteral. De todos los ensayos realizados, sòlo el ensayo de acividad, desarrollado a nivel experimental en animales, no arroja aùn resultados concluyentes. La disponibilidad de esta producciòn en el IICS permitirà desarrollar estudios epidemiològicos acerca de la prevalenciaq de la leishmaniasis en el Paraguay

La escuela como puerta de entrada para el control de la enfermedad de Chagas en las comunidades: una experiencia dentro de un proyecto de mejoramiento de la vivienda / Annual reports 1993 / The rural school as an entrance gate to communities for Chagas disease control an experience inside a housing improvement program

Se considera que la enfermedad de Chagas es el resultado de la interacciòn entre el paràsito (T.cruzi), los vectores triatominos y los huèspedes que viven en condiciones socioeconòmicas de pobreza. La interrupciòn del la transmisiòn vectorial se basa en el rociamiento con insecticidas, el mejoramiento de las viviendas, los programas de educaciòn sanitaria y la participaciòn comunitaria. Sin embargo, la evaluaciòn de la efectividad de tales medidas ha sido realizada aisladamente, lo que les resta valor conclusivo. En el presente estudio se evaluò la capacidad de los niños, que concurren a la escuela de una comunidad donde se desarrollò un proyecto de control de la enfermedad de Chagas, para comprender aspectos bàsicos de la enfermedad asì como para reconocer rastros de triatominos. El èxito de los niños en ambas evaluaciones justifica la incorporaciòn de informaciòn sobre la enfermedad y su control en los programas educativos regulares de zonas endèmicas a fin de preparar a los niños para un eficaz monitoreo de intervenciones de control

Acción tripanocida y toxicidad de alcaloides bisbenzylisouinqlicos y de un derivado fenolico (FC.108), en ensayos in vivo en ratones c3h infectados con T.cruzi experimentalmente / Annual reports 1994

Un total de sesenta ratones C3H infectados con la cepa epsilon, fueron tratados con alcaloides bisbenzylisoquinolicos (50 ratones) y un derivado fenólico (Fc.108), (10 ratones) con el propósito de determinar la actividad tripanocida in vivo contra el T.cruzi de estas sustancias. También se evaluaron otros efectos como: (actividad protectiva y/o toxicidad) al comparar las lesiones del miocardio por medio de estudios histopatológicos en los ratones tratados y no tratados con estos alcaloides. Se encontraron lesiones histopatogicas en todos los ratones tratados y no tratados pero sólo en estos últimos (grupos controles), se hallaron daños mas severos y nidos de amastigotes de T. cruzi en el miocardio. Se encontraron marcadas diferencias entre el grupo control y el grupo de ratones tratados con cycleanine y curine. Todos los ratones tratados presentaron menor parasitemia y una mayor sobrevida

Caracterización de cepas Paraguayas de Trypanosoma Cruzi aislada de pacientes agudos de la enfermedad de Chagas / Characterization of paraguayan Trypanosoma Cruzi strains isolated from acute patients of Chagas disease / Annual Reports 1992

Six Trypanosoma Cruzi Paraguayan strains, isolated from accute chagasic patients, were studied by their behaviour in Bald/C mice, metacylogenesis, biometry of metaciclic froms and lectin agglutination. Early, intermediate and late parasitemic peaks were observed and the strains did not show high virulence. Metacyclogenesis capacity was not higher than 50 percent in all cases, and there were no big differences inthe biometry and lectin agglutination assays. Although previous schizodeme analysis of these strains showed rather homogeneous profiles, biological feautures were quite heterogeneous

Hemocultivos en monos cebus apella experimentalmente infectados con Trypanosoma cruzi y tratado con espirogermanium / Annual Reports 1991 / Hemoculture in Cebus apella monkeys experimentally infected with Trypanosoma cruzi and treated with spirogermanium

twelve cebus apella monkeys experimentally infected with 3 x 10 sobre 5 T. cruzi trypomastigotes Y strain, and treated with different doses of Spirogermanium and Nifurtimox during the acute phase of the infection, were subjected to hemoculture in order to test the sensitivity of this methd in comparsson to direct parasitemia through hemoconcentration. The 75 percent of positive results obtained with hemocutive shows a high sensitivity when compared with direct parasitemia, which did not yield any positive. This makes hemoculture suitable for long term evaluation of potential trypanocide drugs