ABSTRACT
Despite recent advances in the treatment of some forms of leishmaniasis, the available drugs are still far from ideal due to inefficacy, parasite resistance, toxicity and cost. The wide-spectrum antimicrobial activity of 2-nitrovinylfuran compounds has been described, as has their activity against Trichomonas vaginalis and other protozoa. Thus, the aim of this study was to test the antileishmanial activities of six 2-nitrovinylfurans in vitro and in a murine model of leishmaniasis. Minimum parasiticide concentration (MPC) and 50% inhibitory concentration (IC50) values for these compounds against the promastigotes of Leishmania amazonensis, Leishmania infantum and Leishmania braziliensis were determined, as were the efficacies of two selected compounds in an experimental model of cutaneous leishmaniasis (CL) caused by L. amazonensis in BALB/c mice. All of the compounds were active against the promastigotes of the three Leishmania species tested. IC50 and MPC values were in the ranges of 0.8-4.7 µM and 1.7-32 µM, respectively. The compounds 2-bromo-5-(2-bromo-2-nitrovinyl)-furan (furvina) and 2-bromo-5-(2-methyl-2-nitrovinyl)-furan (UC245) also reduced lesion growth in vivo at a magnitude comparable to or higher than that achieved by amphotericin B treatment. The results demonstrate the potential of this class of compounds as antileishmanial agents and support the clinical testing of Dermofural(r) (a furvina-containing antifungal ointment) for the treatment of CL.
Subject(s)
Antiprotozoal Agents/administration & dosage , Cell Proliferation/drug effects , Furans/administration & dosage , Leishmania/drug effects , Leishmaniasis, Cutaneous/drug therapy , Amphotericin B/administration & dosage , Animals , Clinical Trials as Topic , Disease Models, Animal , Female , Humans , In Vitro Techniques , Inhibitory Concentration 50 , KB Cells/drug effects , Leishmania/classification , Leishmania/growth & development , Mice, Inbred BALB C , Neglected Diseases/drug therapy , Time Factors , Vinyl Compounds/administration & dosageABSTRACT
Despite recent advances in the treatment of some forms of leishmaniasis, the available drugs are still far from ideal due to inefficacy, parasite resistance, toxicity and cost. The wide-spectrum antimicrobial activity of 2-nitrovinylfuran compounds has been described, as has their activity against Trichomonas vaginalis and other protozoa. Thus, the aim of this study was to test the antileishmanial activities of six 2-nitrovinylfurans in vitro and in a murine model of leishmaniasis. Minimum parasiticide concentration (MPC) and 50% inhibitory concentration (IC50) values for these compounds against the promastigotes of Leishmania amazonensis, Leishmania infantum and Leishmania braziliensis were determined, as were the efficacies of two selected compounds in an experimental model of cutaneous leishmaniasis (CL) caused by L. amazonensis in BALB/c mice. All of the compounds were active against the promastigotes of the three Leishmania species tested. IC50 and MPC values were in the ranges of 0.8-4.7 µM and 1.7-32 µM, respectively. The compounds 2-bromo-5-(2-bromo-2-nitrovinyl)-furan (furvina) and 2-bromo-5-(2-methyl-2-nitrovinyl)-furan (UC245) also reduced lesion growth in vivo at a magnitude comparable to or higher than that achieved by amphotericin B treatment. The results demonstrate the potential of this class of compounds as antileishmanial agents and support the clinical testing of Dermofural(r) (a furvina-containing antifungal ointment) for the treatment of CL.
Subject(s)
Humans , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Hodgkin Disease/drug therapy , Hodgkin Disease/pathology , Antineoplastic Combined Chemotherapy Protocols/adverse effects , Bleomycin/adverse effects , Bleomycin/therapeutic use , Combined Modality Therapy , Decision Making , Dacarbazine/adverse effects , Dacarbazine/therapeutic use , Doxorubicin/adverse effects , Doxorubicin/therapeutic use , Hodgkin Disease/mortality , Neoplasm Staging , Practice Guidelines as Topic , Risk Assessment , Treatment Outcome , Vinblastine/adverse effects , Vinblastine/therapeutic useABSTRACT
Leishmania es un protozoo parásito causante de la leishmaniasis, enfermedad de variada presentación clínica y de amplia distribución mundial. La Organización Mundial de la Salud la considera una enfermedad re-emergente y no controlada, y sus patrones de transmisión se han visto afectados en los últimos años por la acción humana, entre otros aspectos. El diagnóstico varía de acuerdo con la forma clínica de presentación y actualmente se recomienda la identificación de la especie infectante como elemento de mucha utilidad para indicar un tratamiento adecuado, realizar el monitoreo clínico y como aspecto importante en estudios epidemiológicos, que incluyan el estudio de vectores y/o reservorios. El tratamiento oportuno es, hasta el momento, una de las pocas medidas de control disponibles, ya que a pesar de los esfuerzos realizados, no existe vacuna contra esta afección. En este trabajo se presenta una revisión de la literatura que incluye aspectos importantes de la leishmaniasis, en el contexto internacional actual(AU)
Leishmania is a parasitic protozoon causing Leishmaniasis, disease with a varied clinic presentation and a wide world distribution. WHO considers it as a non-controlled and re-emergent disease and its transmission patterns have been affected in past years by the human action among other features. Its diagnosis change according to the clinical way of presentation and nowadays it is recommended the identification of infectious species like a very useful feature to prescribe a appropriate treatment, to perform the clinical monitoring and the most important in epidemiologic studies including the vector and or reservoirs study. The timely treatment is until now one of the few available control measures since despite the efforts performed there isn't a vaccine against this affection. The aim of present paper is to present a literature review including significant features of Leishmaniasis in the present international context(AU)
Subject(s)
Leishmaniasis , Leishmania/parasitology , CubaABSTRACT
Leishmania es un protozoo parásito causante de la leishmaniasis, enfermedad de variada presentación clínica y de amplia distribución mundial. La Organización Mundial de la Salud la considera una enfermedad re-emergente y no controlada, y sus patrones de transmisión se han visto afectados en los últimos años por la acción humana, entre otros aspectos. El diagnóstico varía de acuerdo con la forma clínica de presentación y actualmente se recomienda la identificación de la especie infectante como elemento de mucha utilidad para indicar un tratamiento adecuado, realizar el monitoreo clínico y como aspecto importante en estudios epidemiológicos, que incluyan el estudio de vectores y/o reservorios. El tratamiento oportuno es, hasta el momento, una de las pocas medidas de control disponibles, ya que a pesar de los esfuerzos realizados, no existe vacuna contra esta afección. En este trabajo se presenta una revisión de la literatura que incluye aspectos importantes de la leishmaniasis, en el contexto internacional actual.
Leishmania is a parasitic protozoon causing Leishmaniasis, disease with a varied clinic presentation and a wide world distribution. WHO considers it as a non-controlled and re-emergent disease and its transmission patterns have been affected in past years by the human action among other features. Its diagnosis change according to the clinical way of presentation and nowadays it is recommended the identification of infectious species like a very useful feature to prescribe a appropriate treatment, to perform the clinical monitoring and the most important in epidemiologic studies including the vector and or reservoirs study. The timely treatment is until now one of the few available control measures since despite the efforts performed there isn't a vaccine against this affection. The aim of present paper is to present a literature review including significant features of Leishmaniasis in the present international context.
ABSTRACT
Leishmania panamensis and Leishmania guyanensis are two species of the subgenus Viannia that are genetically very similar. Both parasites are usually associated with cutaneous leishmaniasis, but also have the potential to cause the mucocutaneous form of the disease. In addition, the study of foci and consequently the identification of vectors and probable reservoirs involved in transmission require a correct differentiation between both species, which is important at epidemiological level. We explored the possibility of identifying these species by using restriction fragment length polymorphisms (RFLP) in the gene coding for heat-shock protein 70 (hsp70). Previously, an hsp70 PCR-RFLP assay proved to be very effective in differentiating other Leishmania species when HaeIII is used as restriction enzyme. Based on hsp70 sequences analysis, BccI was found to generate species-specific fragments that can easily be recognized by agarose gel electrophoresis. Using the analysis of biopsies, scrapings, and parasite isolates previously grouped in a cluster comprising both L. panamensis and L. guyanensis, we showed that our approach allowed differentiation of both entities. This offers the possibility not only for identification of parasites in biological samples, but also to apply molecular epidemiology in certain countries of the New World, where several Leishmania species could coexist.
Subject(s)
HSP70 Heat-Shock Proteins/genetics , Leishmania guyanensis/genetics , Polymerase Chain Reaction/methods , Humans , Leishmania guyanensis/classification , Leishmaniasis, Cutaneous/genetics , Leishmaniasis, Cutaneous/parasitology , Polymorphism, Restriction Fragment Length , Sequence Analysis, DNA , Species SpecificityABSTRACT
In this study, a methanolic extract from Echinaster (Othilia) echinophorus was evaluated for activity against Leishmania amazonensis. The extract showed activity against the promastigote and amastigote forms with IC50 values of 62,9 and 37,5 ìg.mL-1 respectively. This extract showed a moderate toxicity on macrophages from BALB/c mice. A dose of 100 mg/kg/day was effective when administered during 15 days by intraperitoneal route to BALB/c mice infected experimentally...(AU)
Neste estudo descreve-se o efeito de um extrato metanólico de Echinaster echinophorus spp. no parasita Leishmania amazonensis. Em testes com as formas promastigotas e amastigotas, o IC50 do extrato foi 62,9 e 37,5 μg.mL-1, respectivamente. O extrato também tem toxicidade moderada em macrófagos de camundongos BALB/c. O tratamento de camundongos BALB/c infectados com L. amazonensis com doses diárias de 100 mg/kg/dia via intraperitoneal durante 15 dias mostrou-se relativamente efetivo no controle da infecção. Esta investigação confirma a importância de produtos naturais como fonte para a descoberta de fármacos com funções anti-Leishmania(AU)
Subject(s)
Humans , Leishmania mexicana , Antiprotozoal AgentsABSTRACT
INTRODUCCIÓN: la inducción de las proteínas de choque térmico constituyen un mecanismo homeostático que protege a las células del efecto destructivo del calor u otras condiciones de estrés ambiental, paralelamente, ellas cumplen importantes funciones celulares. La proteína de choque térmico de 20 kDa se reportó recientemente en Leishmania amazonensis. OBJETIVO: describir la metodología utilizada para realizar el clonaje de las proteínas de choque térmico, lo que permitió acometer estudios de algunas propiedades biológicas. MÉTODOS: la región codificante del gen hsp20 se amplificó mediante la reacción en cadena de la polimerasa con cebadores adecuados. El producto amplificado se clonó inicialmente en el vector pCR2.1 (Invitrogen) y después en el vector de expresión en procariotas pET-28b (Novagen), para obtener proteína recombinante. De manera paralela, el mismo fragmento se clonó en el vector de expresión en eucariotas pcDNA3 (Invitrogen) para obtener un posible preparado vacunal de ADN. Se realizó la secuenciación nucleotídica de los clones obtenidos, con la finalidad de verificar su fidelidad. RESULTADOS: se obtuvieron plásmidos recombinantes que codifican la HSP20 de Leishmania, y permiten la obtención de proteína recombinante y de ADN en forma masiva. CONCLUSIONES: ambos plásmidos fueron útiles para estudiar algunas de las propiedades biológicas de esta proteína. Este acercamiento puede ser de interés en otros trabajos de esta índole y constituir una guía metodológica(AU)
INTRODUCTION: the induction of heat shock proteins is a homeostatic mechanism that protects cells from the deleterious effects of thermal and other environmental stresses. In addition, they have important cell functions. The 20kDa heat shock protein in Leishmania amazonensis was recently reported. OBJECTIVE: to describe the methodology used for cloning of heat shock proteins, which allowed the study of some biological properties. METHODS: the hsp20 gene coding region was amplified by polymerase chain reaction using adequate primers. The amplified product was initially cloned in pCR2.1 vector (Invitrogen) and then in pET-28b vector (Novagen), to obtain recombinant protein. The same fragment was cloned also in the eukariote expression vector pcDNA3 (Invitrogen). The nucleotidic sequencing of the different clones was made, in order to verify their fidelity. RESULTS: the recombinant plasmids that encode HSP20 protein in Leishmania and allow obtaining massively recombinant protein and DNA were produced. CONCLUSIONS: both plasmids were useful to study some of the biological properties of this protein. This approach could be useful for similar research and represent a suitable methodological guideline(AU)
Subject(s)
Leishmania mexicana/parasitology , Genetic Vectors , Heat-Shock ProteinsABSTRACT
INTRODUCCIÓN: la inducción de las proteínas de choque térmico constituyen un mecanismo homeostático que protege a las células del efecto destructivo del calor u otras condiciones de estrés ambiental, paralelamente, ellas cumplen importantes funciones celulares. La proteína de choque térmico de 20 kDa se reportó recientemente en Leishmania amazonensis. OBJETIVO: describir la metodología utilizada para realizar el clonaje de las proteínas de choque térmico, lo que permitió acometer estudios de algunas propiedades biológicas. MÉTODOS: la región codificante del gen hsp20 se amplificó mediante la reacción en cadena de la polimerasa con cebadores adecuados. El producto amplificado se clonó inicialmente en el vector pCR2.1 (Invitrogen) y después en el vector de expresión en procariotas pET-28b (Novagen), para obtener proteína recombinante. De manera paralela, el mismo fragmento se clonó en el vector de expresión en eucariotas pcDNA3 (Invitrogen) para obtener un posible preparado vacunal de ADN. Se realizó la secuenciación nucleotídica de los clones obtenidos, con la finalidad de verificar su fidelidad. RESULTADOS: se obtuvieron plásmidos recombinantes que codifican la HSP20 de Leishmania, y permiten la obtención de proteína recombinante y de ADN en forma masiva. CONCLUSIONES: ambos plásmidos fueron útiles para estudiar algunas de las propiedades biológicas de esta proteína. Este acercamiento puede ser de interés en otros trabajos de esta índole y constituir una guía metodológica.
INTRODUCTION: the induction of heat shock proteins is a homeostatic mechanism that protects cells from the deleterious effects of thermal and other environmental stresses. In addition, they have important cell functions. The 20kDa heat shock protein in Leishmania amazonensis was recently reported. OBJECTIVE: to describe the methodology used for cloning of heat shock proteins, which allowed the study of some biological properties. METHODS: the hsp20 gene coding region was amplified by polymerase chain reaction using adequate primers. The amplified product was initially cloned in pCR2.1 vector (Invitrogen) and then in pET-28b vector (Novagen), to obtain recombinant protein. The same fragment was cloned also in the eukariote expression vector pcDNA3 (Invitrogen). The nucleotidic sequencing of the different clones was made, in order to verify their fidelity. RESULTS: the recombinant plasmids that encode HSP20 protein in Leishmania and allow obtaining massively recombinant protein and DNA were produced. CONCLUSIONS: both plasmids were useful to study some of the biological properties of this protein. This approach could be useful for similar research and represent a suitable methodological guideline.
ABSTRACT
Current therapy for leishmaniasis is not satisfactory. We describe the in vitro antiproliferative effects of new thiadiazine derivatives against Leishmania amazonensis. The compounds were found to be active against the amastigote form of the parasite, inhibiting parasite growing, from 10 to 89%, at a concentration of 100 ng/ml. This activity suggests that thiadiazine derivatives could be considered as potential antileishmanial compounds.
Subject(s)
Antiprotozoal Agents/pharmacology , Leishmania braziliensis/drug effects , Thiadiazines/pharmacology , Animals , Inhibitory Concentration 50 , Mice , Mice, Inbred BALB C , Parasitic Sensitivity TestsABSTRACT
Current therapy for leishmaniasis is not satisfactory. We describe the in vitro antiproliferative effects of new thiadiazine derivatives against Leishmania amazonensis. The compounds were found to be active against the amastigote form of the parasite, inhibiting parasite growing, from 10 to 89 percent, at a concentration of 100 ng/ml. This activity suggests that thiadiazine derivatives could be considered as potential antileishmanial compounds.
Subject(s)
Animals , Mice , Antiprotozoal Agents , Leishmania braziliensis , Thiadiazines , Inhibitory Concentration 50 , Mice, Inbred BALB CABSTRACT
Se construyó una biblioteca genómica de Leishmania amazonensis mediante el vector pcDNA3, con promotor de expresión en células eucariotas, con el objetivo de contribuir a la aplicación de la tecnología de inmunización con ácidos nucleicos en la leishmaniosis. Para demostrar la expresión de la genoteca en el músculo de ratones inmunizados con esta, se realizó la técnica de inmunofluorescencia indirecta. Como anticuerpo primario se utilizó una mezcla de sueros con alto título antileishmania, de una zona donde predomina la infección con L. braziliensis. Se obtuvo una biblioteca con 80 porciento de clones recombinantes. Se demostró la expresión de determinantes antigénicos en el músculo de ratones BALB/c inmunizados, según resultados de la inmunofluorescencia
Subject(s)
Nucleic Acids/immunology , Animals, Laboratory , Fluorescent Antibody Technique, Indirect , Genomic Library , Immunization/methods , Leishmania mexicana , Mice, Inbred A , Mice, Inbred BALB C/immunologyABSTRACT
Se construyó una biblioteca genómica de Leishmania amazonensis mediante el vector pcDNA3, con promotor de expresión en células eucariotas, con el objetivo de contribuir a la aplicación de la tecnología de inmunización con ácidos nucleicos en la leishmaniosis. Para demostrar la expresión de la genoteca en el músculo de ratones inmunizados con esta, se realizó la técnica de inmunofluorescencia indirecta. Como anticuerpo primario se utilizó una mezcla de sueros con alto título antileishmania, de una zona donde predomina la infección con L. braziliensis. Se obtuvo una biblioteca con 80 por ciento de clones recombinantes. Se demostró la expresión de determinantes antigénicos en el músculo de ratones BALB/c inmunizados, según resultados de la inmunofluorescencia(AU)
Subject(s)
Animals , Mice , Immunization/methods , Nucleic Acids/immunology , Genomic Library , Fluorescent Antibody Technique, Indirect , Animals, Laboratory , Mice, Inbred BALB C/immunology , Leishmaniasis/immunology , Leishmania mexicanaABSTRACT
Parasitólogos de diferentes instituciones médicas de Ciudad de La Habana realizaron la validación externa de ENZYMEBA, procedimiento diagnóstico de amebiosis intestinal en el Instituto de Medicina Tropical "Pedro Kourí". Para ello fueron colectadas muestras seriadas de heces de 212 individuos sobre las que se realizó la observación microscópica (prueba de referencia) y el inmunoensayo ENZYMEBA (prueba en validación). ENZYMEBA, comparada con el examen microscópico de heces, mostró satisfactorios índices de sensibilidad y especificidad. No se observaron reacciones cruzadas con muestras de heces en que estaban presentes otros parásitos. Además, si se tiene en cuenta que para el diagnóstico de amebiosis intestinal con ENZYMEBA es suficiente una sola muestra de heces por paciente, este procedimiento pudiera ser útil en estudios de eficacia terapéutica y de prevalencia (AU)