Actividad antiparasitaria in vitro de plantas de la medicina tradicional Tacana sobre Plasmodium faciparum a través del método fluorométrico-SYBR GreenI / In vitro antiparasitic activity from plants of the Tacana traditional medicine on Plasmodiumfaciparum through thefluorometric method-SYBR GreenI

INTRODUCCIÓN: la presencia de compuestos activos en las plantas las posiciona como una fuente alternativa para el descubrimiento de nuevos fármacos. OBJETIVO: realizar la bioprospección de plantas utilizadas en la medicina tradicional tacana frente a cultivos de Plasmodium falciparum. MÉTODOS: se obtuvieron extractos por maceración en etanol a temperatura ambiente, de 31 órganos colectados de 23 plantas, estos fueron evaluados sobre cultivos asincrónicos de la cepa de Plasmodium falciparum resistente a la Cloroquina (FCR3). A los extractos que mostraron actividad antiplasmódica (CI50<20µg/mL), se evaluó la citotoxicidad (DL50) frente a células HeLa y se calculó el índice de selectividad (IS=DL50/CI50). Los extractos que dieron resultados IS>5, fueron seleccionados como promisorios. RESULTADOS: se obtuvieron 3 plantas muy activas (CI50<10µg/mL); 2 moderadamente activas (10µg/mL20µg/mL). De las 9 plantas que presentaron actividad, solo 2 plantas presentaron IS>5. CONCLUSIONES: incorporar los conocimientos del uso tradicional para realizar las evaluaciones biológicas es de mucha ayuda en la selección de plantas con efectos antiplasmódicos.

INTRODUCTION: the presence of active compounds in plants, converts them as an alternative to find new drugs. OBJECTIVE: carry out the bioprospecting of plants used in Tacana traditional medicine against Plasmodium falciparum cultures. METHODS: from the 31 collected organs of 23 plants, raw extracts were obtained by ethanolic maceration at room temperature and these were evaluated on asynchronic cultures of the strain Plasmodium falciparum resistant to Chloroquine (FCR3). The active extracts (IC50<20µg/mL), were evaluated for cytotoxicity (LD50) against HeLa cells and the Selectivity Index (IS=DL50/IC50) was calculated. The extracts that sowed IS>5were selected as promising. RESULTS: a total of 3 species were very active (IC50<10µg/mL); 2 were moderately active (10µg/mL20µg/mL). From the 9 active plants only 2 presented IS>5. CONCLUSIONS: incorporating the traditional knowledge to carry out biological evaluations is very helpful in the selection of plants with antiplasmodial effects.

Aplicación del método bioquímico-colorimétrico para identificar agentes trypanocidas / Application of the biochemical-colorimetric method to identify trypanocidal agents

La tripanosomiasis americana es una enfermedad infecciosa desatendida, causada por el parásito protozoo Trypanosoma cruzi, que no cuenta con tratamiento en la fase crónica de esta enfermedad mortal, uno de los desafíos es encontrar terapias efectivas para esta compleja enfermedad, dado que no presenta síntomas asociables a la parasitosis por lo que es desconocida entre los médicos tradicionales. Nuestra Facultad está evaluando la medicina tradicional tacana como fuente de agentes antiparasitarios potenciales. El objetivo de este trabajo fue identificar productos naturales trypanocidas utilizando el método colorimétrico XTT-PMS. Para ello, se realizaron curvas de crecimiento de epimastigotes de T. cruzi y determinamos el tiempo óptimo de realización de los ensayos. Se seleccionó la población inicial de trabajo (3x106 parásitos/mL), las condiciones de incubación (medio LIT, 27ºC, 72 horas) y revelado (XTT-PMS, 4 horas). Con el protocolo optimizado, se realizaron evaluaciones de actividad de drogas control, controles naturales y 20 extractos crudos de plantas medicinales de la amazonía. La actividad se basó en cálculos de concentración inhibitoria media y se consideraron activos las sustancias con CI50<50µg/mL. De los 20 extractos evaluados, el 40% fueron activos. Las plantas más interesantes fueron Sipu sipu (CI50=8.9±1.7µg/mL), Ejije bid'u (CI50=9.1±1.5µg/mL) e Id'ene eidhue (CI50=10.8±1.1µg/mL) con valores de CI50 cercanos a los controles, confirmando la utilidad y potencial del protocolo desarrollado

American trypanosomiasis is listed among the unattended infectious disease, is caused by the protozoan parasite Trypanosoma cruzi, and has no treatment in the chronic phase of this deadly disease. One of the challenges is finding effective therapies for this complex disease, given that it does not present any associated symptoms to the parasitism and is unknown among traditional doctors. Our Faculty is evaluating tacana traditional medicine as a source of potential antiparasitic agents. The objective of this work was to identify trypanocidal natural products using the XTTPMS colorimetric method. For this, growth curves of T. cruzi epimastigotes were made to determine the optimal time to carry out the tests. The initial work population (3x106 parasites/mL), the incubation conditions (medium LIT, 27ºC, 72 hours) and revealed process (XTT-PMS, 4 hours) were selected. With the optimized protocol, activity evaluations of control drugs, natural controls and 20 crude extracts of medicinal plants of the Amazon were carried out. The activity was based on calculations of mean inhibitory concentration and substances with IC50 <50µg/mL were considered active. Of the 20 extracts evaluated, 40% were active. The most interesting plants were Sipu sipu (IC50=8.9±1.7µg/mL), Ejije bid'u (IC50=9.1±1.5µg/mL) and Id'ene eidhue (IC50=10.8±1.1µg/mL) with values of IC50 close to the controls, confirming the usefulness and potential of the developed protocol.

Desparasitación en niños de escuelas rurales 2016-2017: diagnóstico y tratamiento de enteroparásitos intestinales en la escuela de Sapecho A ­ Sud Yungas, La Paz / Deworming in children of rural schools 2016-2017: diagnosis and treatment of intestinal enterocoparasites in Sapecho A School - Sud Yungas, La Paz

Los parásitos intestinales son un grave problema de salud pública donde la alta prevalencia está asociado a la falta de educación sanitaria, hábitos higiénicos e infraestructura inadecuada. El presente trabajo es un estudio descriptivo de corte transversal, donde el universo de trabajo son todos los niños de la Escuela Sapecho A (Gestión 2016-2017), trabajo que involucro a estudiantes de pregrado y de post grado de la Facultad de Ciencia Farmacéuticas y Bioquímicas, donde a través de estudios coproparasitológicos (técnica de Ritchie) se pudo observar que la mayoría de la población, entre 78,24 y 88,4%, se encontraba infectada por uno o más parásitos (Helmintos­Protozoos). Luego de realizar una desparasitación masiva con albendazol (400mg/Dos dosis) se llegó a una reducción respecto a los helmintos de 53,5 y 65,2% respectivamente en cada gestión, sin embargo, este no fue efectivo contra los protozoos. Para alcanzar un éxito en el tratamiento se debe tratar al grupo familiar y dar énfasis al componente educativo de higiene y limpieza.

Intestinal parasites are a serious public health problem where the high prevalence is associated with a lack of education, inadequate hygienic habits and sanitary infrastructure. The present work is a cross-sectional descriptive study, where the universe of work are the children of Sapecho A School (Management 2016-2017), work that involved undergraduate and post-graduate students of the Faculty of Pharmaceutical Science and Biochemistry where through coproparasitological studies (Ritchie's technique) it was observed that the majority of the population, between 78.24 and 88.4%, was infected by one or more parasites (Helminths-Protozoa). After performing a massive deworming treatment with albendazole (400mg/Two doses) a reduction was reached with respect to the helminths of 53.5 and 65.2%, respectively in each year, however this was not effective against the protozoa. To achieve a successful treatment, the family group must be treated and the educational component of hygiene and cleanliness should be emphasized.

Estudio piloto de diagnóstico de parasitosis intestinal y respuesta al uso de Mebendazol en niños de edad escolar en las escuelas Charcas II, El Sillar y la Cascada, gestión 2010 La Paz / Pilot study of intestinal parasitosis diagnostic and response to Mebendazol on school age children at schools in Charcas II, El Sillar and La Cascada, year 2010 La Paz

El proyecto IDH 09: Desparasitación de niños en escuelas rurales", llevo adelante un trabajo piloto experimental de diagnóstico sobre parásitos intestinales en niños en las Escuelas de las Comunidades: Charcas II; La Cascada y El Sillar, Provincia Sud Yungas, Departamento de La Paz, Bolivia. El análisis coproparasitológico fue en 93 muestras tomadas entre Inicial y quinto de primaria, con edades entre 5 y 12 años. En promedio, el 97,9% de las muestras indicaron presencia de Protozoarios y hasta un 54,5% de Helmintos, concomitantemente, con una relación promedio de 2,0 veces más Protozoarios. En las escuelas de Charcas II y La Cascada la relación fue de 1,8 y en El Sillar fue de 2,5. Hasta 12 parásitos fueron identificados entre los Protozoarios: Blatocystis hominis (92,7%); Entamoeba coli (50,3%); Endolimax nana (44,0%); Giardia lamblia (39,3%); Iodamoeba bütschlii (25,0%) y Chilomastix mesnili (8,3%) y entre los Helmintos: Ascaris lumbricoides (30,0%); Uncinaria spp (21,7%), Strongyloides stercoralis (9,0%); Hymenolepis nana (7,0%) y Trichuris trichiura (5,7%), en una muestra se detectó Enterobius vermicularis. En la escuela Charcas II, de acuerdo a sus programas de desparasitación, los niños recibieron tratamiento con Mebendazol y el efecto de la medicación fue evaluado, aleatoriamente, a los 7 días, sobre un total de 21 niños. El Mebendazol (1200mg) elimino 50% de los Helmintos. A. lumbricoides fue eliminado de todas las muestras, Uncinaria spp, S. stercoralisy T. trichiura fueron eliminados en un 50%, mientras que H. nana persistió en todas las muestras, mientras que los Protozoarios fueron eliminados solo en un 19% de las muestras.

The Project Deworming of children in rural schools carried out a pilot experimental field work to determine intestinal parasites levels in kids in rural schools, at Charcas II, La Cascada and El Sillar communities, South Yungas province, Department of La Paz, Bolivia. The coproparasitologic studies were carry out on 93 feces samples, from kids from initial to fifth grade, within ages from 5 to 12 years. An average of 97,9% of the samples showed presence of protozoa parasites and up to 54,5% showed, additionally, presence of Helminthes, with a general ratio of Protozoan to Helminthes of 2,0. At Charcas II School and La Cascada School the ratio was of 1,8; while at El Sillar gave a ratio of 2,5. A total of 12 parasites were identified, among the protozoa: Blatocystishominis (92,7%); Entamoeba coli (50,3%); Endolimax nana (44,0%); Giardia lamblia (39,3%); Iodamoeba bütschlii (25,0%) and Chilomastix mesnili:(8,3%) and among the Helminthes: Ascaris lumbriocoides (30,0%); Uncinaria spp (21,7%), Strongyloides stercoralis (9,0%); Hymenolepis nana (7,0%) and Trichuris trichiura (5,7%) and in one sample we detected Enterobius vermicularis. According to their deworming program, at Charcas II School, kids received treatment with Mebendazol (1200mg) and the effect was evaluated 7 days after treatment. On a total of 21 children. Mebendazol eliminated 50% of Helminthes. A. lumbricoides was eliminated from all samples; Uncinaria spp, S. stercoralis y T. trichiura only from 50% of the samples and H. nana persisted in all samples, while Protozoan parasites were eliminated on nearly 19% of the samples

Estudios de susceptibilidad de cepas de Leishmania aethiopica frente a alcaloides de Galipea longiflora (Evanta) / Susceptibility studies on Leishmania aethiopica strains against total alkaloids from Galipea longiflora (Evanta)

El Instituto de Investigaciones Fármaco Bioquímicas (IIFB), de la Facultad de Ciencias Farmacéuticas y Bioquímicas, de la UMSA, desarrolla trabajos sobre la actividad leishmanicida, de los alcaloides totales (CAT) obtenidos de la corteza de la especie medicinal amazónica conocida como Evanta (Galipea longiflora) por los Pueblos Tacana, Tsimane y Mosetene. Como parte de las actividades del Proyecto UMSA-ASDI Biomoleculas de interés medicinal e industrial (antiparasitarios) hemos podido contar con la estadía, en el IIFB, de un investigador del Armauer Hansen Research Institute (AHRI) de Etiopia, lo que nos ha permitido desarrollar evaluaciones de CAT, Miltefocine y Amfotericina B, frente a cepas de Leishmania aethiopica, agente causante de las diversa formas de Leishmaniais cutánea en Etiopía. Un total de seis cepas, de L. aethiopica, fueron adaptadas a condiciones in vitro y mostraron un comportamiento homogéneo frente a CAT, cinco de estas cepas mostraron un valor promedio de IC50 = 8,68 ±1,56 mg/mL, valor algo inferior a los calculados para nuestras cepas de referencia, L. amazonensis y L. braziliensis con IC50 = 11,73 ± 4,32 mg/mL y IC50 = 12,28 ±- 2,95 mg/mL, respectivamente. Excepto por una cepa de L. aethiopica que mostro valores consistentemente más elevados que el resto con IC50= 14,37 ± 3,58 mg/mL. Como consecuencia de esta interacción científica, la Universidad Mayor de San Andrés (UMSA) ha firmado un Memorandum de Entendimiento para el desarrollo de investigaciones conjuntas, con el Armauer Hansen Research Institute (AHRI), dependiente del Ministerio de Salud de Etiopia y explorar la posibilidad de que nuestra experiencia de validación clínica con Evanta en el tratamiento de leishmaiasis cutánea, en Bolivia, podría ser replicada en Etiopía, donde se reportan entre 20,000 a 30,000 nuevos casos de Leismaniasis por año.

The Instituto de Investigaciones Fármaco Bioquímicas (IIFB), at the Faculty of Pharmaceutical and Biochemical Sciences, from UMSA, carry out work related to the leishmanicidal activity of the total alkaloids (CAT) obtained from the bark of the Amazonian medicinal species known as Evanta (Galipea longiflora) by the Tacana, Tsimane y Mosetene people. As part of the activities develop by the UMSA-ASDI Project Biomolecules of medicinal and industrial Interest (antiparasitic) we had a visit, in our laboratories at IIFB, of a researcher from The Armauer Hansen Research Institute (AHRI) from Ethiopia, during his stay we were able to carry out evaluations of CAT, Miltefocine and Anphotericin B, against strains of L. aethiopica, causative agent of the different manifestations of cutaneous leishmaniasis in Ethiopia. A total of six strains of L. aethiopica, were adapted to in vitro a conditions, at IIFB; and did show homogenous behavior against CAT. Five of the strains, showed an average calculated value for IC50 = 8.68 ±1.56 mg/mL, a value somewhat lower to the calculated for the reference strains L. amazonensis and L. braziliensis with IC50 = 11.73 ± 4.32 mg/mL and IC50 = 12.28 +/- 2.95 mg/mL, respectively. Except for one strain that showed values somewhat higher, to the other strains, consistently through our studies, with IC50 = 14.37 ± 3.58 mg/mL. As a consequence of our scientific interaction, the Universidad Mayor de San Andrés (UMSA) has signed a Memorandum of Understanding for the development of joint research with the Armauer Hansen Research Institute (AHRI) that belongs to the Ministry of Health in Ethiopia, and explore the possibilities to replicate the Bolivian clinical validation experience of Evanta in the treatment of cutaneous leishmaniasis, in Ethiopia where the annual incidence is estimated to be between 20, 000 to 30, 0000.

Antileishmanial metabolites from Lantana balansae

ABSTRACT Eleven compounds, 12-oxo-phytodienoic acid (1), persicogenin (2), eriodictyol 3′,4′,7-trimethyl ether (3), phytol (4), spathulenol (5), 4-hydroxycinnamic acid (6), onopordin (7), 5,8,4′-trihydroxy-7,3′-dimethoxyflavone (8), quercetin (9), jaceosidin (10), and 8-hydroxyluteolin (11), were isolated from an ethanol extract of Lantana balansae Briq., Verbenaceae, that was found to possess antileishmanial activity. The structures of the compounds were determined by NMR spectroscopy and HR mass spectrometry, and 1, 2, 3, 7, 8 and 9 were investigated for antiprotozoal activity toward promastigotes of Leishmania amazonensis and Leishmania braziliensis. Compound 1 was shown to be the most potent, with the IC50 values 2.0 µM toward L. amazonensis and 0.68 µM toward L. braziliensis, although less potent than the positive control Amphotericin B. All compounds have been reported previously, but this is the first report of the isolation of a cyclopentenone fatty acid (1) and flavanones (2 and 3) from a Lantana species.

Drimanes from Drimys brasiliensis with leishmanicidal and antimalarial activity

This paper evaluates CHCl3 and CH3OH extracts of the stem bark, branches and leaves of Drimys brasiliensis and drimane sesquiterpenes isolated from the stem bark against strains of Leishmania amazonensis and Leishmania braziliensis promastigotes and Plasmodium falciparum trophozoites. All of the extracts and compounds were tested in cell lines in comparison with reference standards and cell viability was determined by the XTT method. The CHCl3 and CH3OH extracts from the stem bark and branches yielded promising results against two strains of Leishmania, with 50% inhibitory concentrations (IC50 ) values ranging from 39-100 µg/mL. The CHCl3 extract of the stem bark returned IC50 values of 39 and 40.6 µg/mL for L. amazonensis and L. braziliensis, respectively. The drimanes were relatively effective: 1-β-(p-coumaroyloxy)-polygodial produced IC50 values of 5.55 and 2.52 µM for L. amazonensis and L. braziliensis, respectively, compared with 1-β-(p-methoxycinnamoyl)-polygodial, which produced respective IC50 values of 15.85 and 17.80 µM. The CHCl3 extract demonstrated activity (IC50 of 3.0 µg/mL) against P. falciparum. The IC50 values of 1-β-(p-cumaroyloxyl)-polygodial and 1-β-(p-methoxycinnamoyl)-polygodial were 1.01 and 4.87 µM, respectively, for the trophozoite strain. Therefore, the results suggest that D. brasiliensis is a promising plant from which to obtain new and effective antiparasitic agents.

Chemical composition of essential oils of Piper jacquemontianum and Piper variabile from Guatemala and bioactivity of the dichloromethane and methanol extracts

The essential oils from two native species from Guatemala were studied for their chemical composition and the dichloromethane and methanol extracts for their biological activity. A GC-MS analysis of the essential oil from Piper jacquemontianum Kunth, Piperaceae, showed 34 constituents, consisting mainly of linalool (69.4 percent), while Piper variabile C. DC. essential oil had 36 constituents, camphor (28.4 percent), camphene (16.6 percent) and limonene (13.9 percent) being the major components. Dichloromethane extracts of both species were cytotoxic against MCF-7, H-460 and SF-268 cell lines (<7 µg/mL). Dichloromethane extract of P. jacquemontianum was slightly active against bacteria (0.5 mg/mL), was active against promastigotes of Leishmania (20.4-61.0 µg/mL), and epimastigotes of Trypanosoma cruzi (51.9 µg/mL). The methanol extract of P. variabile showed antimalarial activity against Plasmodium falciparum F32 (4.5 µg/mL), and the dichloromethane extract against Leishmania (55.8-76.3 µg/mL) and T. cruzi (45.8 µg/mL). None of the extracts from the two species was active against Aedes aegypti larvae and Artemia salina nauplii.