Trichilones A-E: New Limonoids from Trichilia adolfi.

In addition to the trichilianones A-D recently reported from Trichilia adolfi, a continuing investigation of the chemical constituents of the ethanol extract of the bark of this medicinal plant yielded the five new limonoids 1-5. They are characterized by having four fused rings and are new examples of prieurianin-type limonoids, having a ε-lactone which in 4 and 5 is α, ß- unsaturated. The structures of the isolated metabolites were determined by high field NMR spectroscopy and HR mass spectrometry. The new metabolites were shown to have the ε-lactone fused with a tetrahydrofuran ring which is connected to an oxidized hexane ring joined with a cyclo-pentanone having a 3-furanyl substituent. As the crude extract possesses antileishmanial activity, the compounds were assayed for cytotoxic and antiparasitic activities in vitro in murine macrophage cells (raw 264.7 cells) and in Leishmania amazoniensis as well as L. braziliensis promastigotes. Metabolites 1-3 and 5 showed moderate cytotoxicity (between 30-94 µg/mL) but are not responsible for the antileishmanial effect of the extract.

Trichilianones A-D, Novel Cyclopropane-Type Limonoids from Trichilia adolfi.

The fractionation of an ethanol extract of the bark of Trichilia adolfi yielded four novel limonoids (trichilinones A-D, 1-4), with five fused rings and related to the hortiolide-type limonoids. Starting with an ε-lactone, which is α,ß-unsaturated in trichilinones A and D (1 and 4), attached to a tetrahydrofuran ring that is connected to an unusual bicyclo [5.1.0] hexane system, joined with a cyclopentanone with a 3-furanyl substituent [(2-oxo)-furan-(5H)-3-yl in trichilinone D (4)], the four compounds isolated display a new 7/5/3/5/5 limonoid ring system. Their structures were established based on extensive analysis of NMR spectroscopic data. As the crude extract possessed anti-leishmanial properties, the compounds were assayed for cytotoxic and anti-parasitic activities in vitro in murine macrophages cells (Raw 264.7) and leishmania promastigotes (L. amazoniensis and L. braziliensis), respectively. The compounds showed moderate cytotoxicity (approximately 70 µg/mL), but are not responsible for the leishmanicidal effect of the extract.

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

Leishmanicidal and cytotoxic activity from plants used in Tacana traditional medicine (Bolivia).

ETNOPHARMACOLOGICAL RELEVANCE: Thirty-eight Tacana medicinal plant species used to treat skin problems, including leishmania ulcers, skin infections, inflammation and wound healing, were collected in the community of Buena Vista, Bolivia, with the Tacana people. Twenty two species are documented for the first time as medicinal plants for this ethnic group living in the northern area of the Department of La Paz. AIM OF THE STUDY: To evaluate the leishmanicidal effect (IC50) and cytotoxicity (LD50) of the selected plants. To carry out bioguided studies on the active extracts. To assess the potential of Bolivian plant biodiversity associated with traditional knowledge in the discovery of alternative sources to fight leishmaniasis. MATERIALS AND METHODS: Seventy three ethanol extracts were prepared from 38 species by maceration and were evaluated in vitro against promastigotes of Leishmania amazonensis and L. braziliensis. Active extracts (IC50 ≤ 50 µg/mL) were fractionated by chromatography on Silica gel column and the fractions were assessed against the two Leishmania strains. The most active fractions and the crude extracts were evaluated against reference strains of L. amazonensis, L. braziliensis, L. aethiopica, two native strains (L. Lainsoni and L. braziliensis) and for cytotoxicity against HeLa cells. The chromatographic profile of the active fractions was obtained by reverse phase chromatography using HPLC. RESULTS: From the 73 extracts, 39 extracts (53.4%) were inactive and 34 showed activity. Thirteen species were sselected for bioguided studies. The crude extracts and their 36 fractions were evaluated against two Leishmania strains. The most active fraction were tested in a panel of five leishmania strains and for cytotoxicity. The Selective Index (SI = LD50/IC50) was calculated, and were generally low. Retention time and UV spectra were recorded for the active fractions by HPLC-DAD using a reverse phase column. Profiles were very different from each other, showing the presence of different compounds. CONCLUSION: Bolivian traditional knowledge from the Tacanba was useful to identify plants with effect on Leishmania promastigotes. Chromatographic bioguided studies showed stronger leishmanicidal and cytotoxic activity for the medium polar fraction. HPLC analysis showed different chromatographic profiles of the active fractions.