Antigenotoxic Effect of Ascorbic Acid and Resveratrol in Erythrocytes of Ambystoma mexicanum, Oreochromis niloticus and Human Lymphocytes Exposed to Glyphosate.

Glyphosate is a controversial herbicide. Its genotoxicity and presence in various ecosystems have been reported. The use of ascorbic acid and resveratrol could protect different organisms from glyphosate-induced genetic damage. In the present study, specific genetic damage induced by glyphosate was evaluated in erythrocytes of Oreochromis niloticus, Ambystoma mexicanum and human lymphocytes. Simultaneously, the antigenotoxic capacity of various concentrations of ascorbic acid and resveratrol was evaluated by means of pretreatment and simultaneous treatment protocols. The 0.03, 0.05 and 0.07 mM concentrations of glyphosate induced significant genotoxic activity (p < 0.05) in human lymphocytes and in erythrocytes of the species studied, and could cause genomic instability in these populations. The reduction in genetic damage observed in human lymphocytes exposed to high concentrations of glyphosate is only apparent: excessive genetic damage was associated with undetectable excessive tail migration length. A significant (p < 0.05) antigenotoxic effect of ascorbic acid and resveratrol was observed in all concentrations, organisms and protocols used. Both ascorbic acid and resveratrol play an important role in maintaining the integrity of DNA. Ascorbic acid in Oreochromis niloticus, Ambystoma mexicanum reduced glyphosate-induced genetic damage to a basal level. Therefore, our data indicate that these antioxidants could help preserve the integrity of the DNA of organisms exposed to glyphosate. The consumption of antioxidants is a useful tool against the genotoxicity of glyphosate.

Entrenamiento de sprint mediante el uso de trineo y paracaídas

El entrenamiento individualizado de carreras específicas con medios resistidos es una importante herramienta para la mejora de la velocidad. En virtud a la demanda de esta capacidad para el buen desempeño de los jugadores de béisbol, se reconoce como objetivo del presente estudio diseñar un entrenamiento de sprint mediante el uso de trineo y paracaídas. Se planifica un cuasiexperimento, con dos grupos: control y experimental y en dos momentos: pre y postest. La etapa experimental se desarrolla durante la pretemporada, conformada por una muestra de diez sujetos con 20.84 años de edad y 79.82 kg. de peso promedio. Se emplearon como métodos teóricos el analítico-sintético, inductivo-deductivo, histórico-lógico, sistémico-estructural-funcional y como empíricos el análisis de contenido, la observación, la medición. Para la medición de la velocidad lineal, las variables analizadas son el test de 60 yardas y los test de fuerza máxima del trineo y de squat. Los resultados alcanzados indicaron mejoras significativas en el grupo experimental en los tres test realizados, con % de incrementos iguales a 3.48, 7.25 y 7.46 % respectivamente. Además, se obtiene que existe una elevada correlación entre la fuerza máxima del trineo y la de squat con respecto al peso de rendimiento al esfuerzo de los atletas, con coeficientes de Pearson iguales a 63.6 % y 62.9 % respectivamente y para un 95 % de confianza. Se demuestra que el entrenamiento resistido nos proporciona información clave en la fase de velocidad máxima para la mejora del rendimiento de sprint en el béisbol.

O treinamento individualizado de corridas específicas com mídia resistida é uma ferramenta importante para melhorar a velocidade. Em virtude da demanda desta capacidade para o bom desempenho dos jogadores de beisebol, é reconhecido como um objetivo do presente estudo projetar um treinamento de sprint usando trenó e pára-quedas. Está prevista uma quase-experimentação, com dois grupos: controle e experimental e em dois momentos: pré-teste e pós-teste. A fase experimental ocorreu durante a pré-época, com uma amostra de dez sujeitos de 20,84 anos de idade e pesando uma média de 79,82 kg. Os métodos teóricos utilizados foram analítico-sintético, indutivo-dedutivo, histórico-lógico, sistêmico-estrutural-funcional, e os métodos empíricos foram análise de conteúdo, observação e medição. Para a medição da velocidade linear, as variáveis analisadas foram o teste das 60 jardas e os testes de resistência máxima do trenó e do agachamento. Os resultados alcançados indicaram melhorias significativas no grupo experimental nos três testes realizados, com aumentos de % iguais a 3,48, 7,25 e 7,46 %, respectivamente. Além disso, obtém-se que existe uma alta correlação entre a força máxima do trenó e o agachamento com relação ao peso do desempenho de esforço dos atletas, com coeficientes de Pearson iguais a 63,6 % e 62,9 % respectivamente e para um 95 % de confiança. Demonstra-se que o treinamento resistido nos fornece informações chave na fase de velocidade máxima para a melhoria do desempenho do sprint no beisebol.

The individualized training of specific races with resisted means is an important tool for the improvement of speed. Due to the demand of this capacity for the good performance of baseball players, the objective of this study is to design a sprint training using sled and parachute. A quasi-experiment is planned, with two groups: control and experimental and in two moments: pre-test and post-test. The experimental stage is developed during the pre-season, conformed by a sample of ten subjects with 20.84 years of age and 79 82kg. of average weight. Theoretical methods used were analytical synthetic, inductive-deductive, historical-logical and systemic-structural-functional. The empirical methods were content analysis, observation and measurement. For the measurement of linear speed, the variables analyzed are the 60-yard test and the maximum strength tests of the sled and squat. The results achieved indicated significant improvements in the experimental group in the three tests performed, with % increases equal to 3.48, 7.25 and 7.46 respectively. In addition, it is obtained that there is a high correlation between the maximum strength of the sled and the squat with respect to the weight of the athletes' effort performance, with Pearson coefficients equal to 63.6 % and 62.9 % respectively and for a 95 % of confidence. It is demonstrated that resisted training provides key information in the maximum velocity phase for the improvement of sprint performance in baseball.

Ecopathogenic complexes of American trypanosomiasis in endemic areas of Venezuela: Diagnosis and variability of Trypanosoma cruzi.

BACKGROUND & OBJECTIVES: Trypanosoma cruzi, the causative agent of American trypanosomiasis, has been reported in 180 mammalian species and 154 triatomines species of Neotropic. This is a clonal parasite with variable biological behaviour, associated with the genetics of the parasite and its hosts. To know the eco-pathogenic complex of this zoonosis, it was proposed to characterize T. cruzi isolates obtained from triatomines and domestic, peridomestic and wild mammals of the eastern and central-western regions of Venezuela. METHODS: The positivity to T. cruzi was established and the isolates were genetically characterized by PCR amplification of the mini-exon gene, the DNA coding for 24Sa and 18S rRNA, and polymorphic sequences-RFLPs. The sampling sites were georeferenced using the MapSource Software and ArcGis 9.3 programs to generate distribution maps of the isolates. RESULTS: Of the 460 hosts (205 triatomines and 255 mammals), 49% were positive for the parasite. On the other hand, 38 isolates obtained from the triatomines and 23 isolates obtained from mammals were evaluated. The TcI genotype predominated in most of the isolates; however, in those obtained from triatomines the presence of the TcIII genotype in single infections and TcI + TcIII or TcI + TcIV in mixed infections was also evidenced. INTERPRETATION & CONCLUSION: There is a possibility that the triatomines act as biological syringes for these genotypes associated exclusively to them. The heterogeneity in T. cruzi isolates demonstrated the complexity of parasitosis in these regions, presenting its control and prevention as a challenge.

Programa de ejercicios con bandas elásticas de resistencia para el incremento de la velocidad en la carrera home-primera base, con jugadores de béisbol, categoría juvenil de Matanzas / Program of exercises with elastic resistance bands for the increase of the speed in the home-first base race with junior baseball players from Matanzas

RESUMEN En las últimas décadas, se han incorporado a los programas de entrenamiento ejercicios con la utilización de tubos (redondos) o bandas (planas) elásticas, de resistencia progresiva, lo que constituye una herramienta fundamental para desarrollar la fuerza muscular e imitar movimientos o gestos deportivos en numerosas especialidades deportivas, debido a su facilidad de uso. El objetivo del presente estudio fue diseñar un programa de entrenamiento con bandas elásticas de resistencia para el incremento de la velocidad, en la carrera home-primera base, con jugadores de la categoría juvenil de Matanzas que participaron en el XLVI Campeonato Nacional de Béisbol. La etapa experimental se desarrolla durante el periodo preparatorio, que constó de 20 microciclos de duración, con una frecuencia de tres días por semana, utilizando la combinación de ejercicios de velocidad con bandas o tubos elásticos y los modelos de periodización de la fuerza y el Sistema de Bloque o Fuerza Concentrada. Los resultados demuestran cambios significativos en los tiempos de la carrera home-primera base de los jugadores, en los diferentes controles efectuados para un 95 % de confianza, pues el valor de la probabilidad (p-value=0.0001) es menor que 0.05. Además, se alcanza un 4.37 % de incremento, con una reducción de los tiempos de 0.37 segundos.

ABSTRACT In the last decades, exercises with the use of tubes (round) or elastic bands (flat) of progressive resistance have been incorporated into the training programs, being a fundamental tool to develop muscle strength and imitate sport movements or gestures in numerous sports specialties, because of its ease to be used. The objective of this study was to design a training program with elastic resistance bands for the increase of the speed in the home-first base race with junior baseball players from Matanzas who participated in the XLVI National Baseball Championship. The experimental stage is developed during the preparatory period, which had a duration of 20 micro cycles, with a frequency of three days per week, using the combination of speed exercises with bands or elastic tubes and the models of periodization of strength and the System of Block or Concentrated Strength. The results show significant changes in the times of the home-first base race of the players in the different controls carried out with a 95% of confidence, since the value of the probability (p-value=0.0001) is less than 0.05. In addition, a 4.37 % increase was achieved with a reduction of 0.37 seconds.

HaloSeeker 1.0: A User-Friendly Software to Highlight Halogenated Chemicals in Nontargeted High-Resolution Mass Spectrometry Data Sets.

In the present work, we address the issue of nontargeted screening of organohalogenated chemicals in complex matrixes. A global strategy aiming to seek halogenated signatures in full-scan high-resolution mass spectrometry (HRMS) fingerprints was developed. The resulting all-in-one user-friendly application, HaloSeeker 1.0, was developed to promote the accessibility of associated in-house bioinformatics tools to a large audience. The ergonomic web user interface avoids any interactions with the coding component while allowing interactions with the data, including peak detection (features), deconvolution, and comprehensive accompanying manual review for chemical formula assignment. HaloSeeker 1.0 was successfully applied to a marine sediment HRMS data set acquired on a liquid chromatography-heated electrospray ionization [LC-HESI(-)] Orbitrap instrument ( R = 140 000 at m/z 200). Among the 4532 detected features, 827 were paired and filtered in 165 polyhalogenated clusters. HaloSeeker was also compared to three similar tools and showed the best performances. HaloSeeker's ability to filter and investigate halogenated signals was demonstrated and illustrated by a potential homologue series with C12H xBr yCl zO2 as a putative general formula.

In ovo transformation of two emerging flame retardants in Japanese quail (Coturnix japonica).

Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) and Dechlorane Plus (DP) are two chlorinated, alternative flame retardants that have been found in wild birds and bird eggs. Little is known about the fate and effect of these compounds in birds, especially during the vulnerable stages of embryonic development. To investigate the ability of birds to biotransform these compounds, an in ovo exposure experiment with Japanese quail eggs was performed. Quail eggs were injected in the yolk sac with 1000ng/g egg of TDCIPP (2.3 nmol/g ww), DP (1.5 nmol/g ww) or a mixture of both and were then incubated at 37.5°C for 17 days. To get a time-integrated understanding of the in ovo transformation of the compounds, one egg per treatment was removed from the incubator every day and analyzed for TDCIPP and its metabolite bis(1,3-dichloro-2-propyl) phosphate (BDCIPP) and/or for DP. By the end of the incubation period, TDCIPP was completely metabolized, while simultaneously BDCIPP was formed. The conversion of the parent compound into the metabolite did not occur proportionally and the concentration of BDCIPP showed a tendency to decrease when TDCIPP became depleted, both indicating that BDCIPP was further transformed into compounds not targeted for analysis. Further untargeted investigations did not show the presence of other metabolites, possibly due to the volatility of the metabolites. On the other hand, the DP concentration did not decrease during egg incubation. This study indicates that within the incubation period, avian embryos are able to biotransform TDCIPP, but not DP.

Screening halogenated environmental contaminants in biota based on isotopic pattern and mass defect provided by high resolution mass spectrometry profiling.

In the present work, we addressed the question of global seeking/screening organohalogenated compounds in a large panel of complex biological matrices, with a particular focus on unknown chemicals that may be considered as potential emerging hazards. A fishing strategy was developed based on untargeted profiling among full scan acquisition datasets provided by high resolution mass spectrometry. Since large datasets arise from such profiling, filtering useful information stands as a central question. In this way, we took advantage of the exact mass differences between Cl and Br isotopes. Indeed, our workflow involved an innovative Visual Basic for Applications script aiming at pairing features according to this mass difference, in order to point out potential organohalogenated clusters, preceded by an automated peak picking step based on the centWave function (xcms package of open access R programming environment). Then, H/Cl-scale mass defect plots were used to visualize the datasets before and after filtering. The filtering script was successfully applied to a dataset generated upon liquid chromatography coupled to ESI(-)-HRMS measurement from one eel muscle extract, allowing for realistic manual investigations of filtered clusters. Starting from 9789 initial obtained features, 1994 features were paired in 589 clusters. Hexabromocyclododecane, chlorinated paraffin series and various other compounds have been identified or tentatively identified, allowing thus broad screening of organohalogenated compounds in this extract. Although realistic, manual review of paired clusters remains time consuming and much effort should be devoted to automation.

Triatoma maculata, the Vector of Trypanosoma cruzi, in Venezuela. Phenotypic and Genotypic Variability as Potential Indicator of Vector Displacement into the Domestic Habitat.

Triatoma maculata is a wild vector of Trypanosoma cruzi, the causative agent of Chagas disease; its incursion in the domestic habitat is scant. In order to establish the possible domestic habitat of T. maculata, we evaluated wing variability and polymorphism of genotypic markers in subpopulations of T. maculata that live in different habitats in Venezuela. As markers, we used the mtCyt b gene, previously apply to evaluate population genetic structure in triatomine species, and the ß-tubulin gene region, a marker employed to study genetic variability in Leishmania subgenera. Adults of T. maculata were captured in the period 2012-2013 at domestic, peridomestic (PD), and wild areas of towns in the Venezuelan states of Anzoátegui, Bolívar, Portuguesa, Monagas, Nueva Esparta, and Sucre. The phenotypic analysis was conducted through the determination of the isometric size and conformation of the left wing of each insect (492 individuals), using the MorphoJ program. Results reveal that insects of the domestic habitat showed significant reductions in wing size and variations in anatomical characteristics associated with flying, in relation to the PD and wild habitats. The largest variability was found in Anzoátegui and Monagas. The genotypic variability was assessed by in silico sequence comparison of the molecular markers and PCR-RFLP assays, demonstrating a marked polymorphism for the markers in insects of the domestic habitat in comparison with the other habitats. The highest polymorphism was found for the ß-tubulin marker with enzymes BamHI and KpnI. Additionally, the infection rate by T. cruzi was higher in Monagas and Sucre (26.8 and 37.0%, respectively), while in domestic habitats the infestation rate was highest in Anzoátegui (22.3%). Results suggest domestic habitat colonization by T. maculata that in epidemiological terms, coupled with the presence in this habitat of nymphs of the vector, represents a high risk of transmission of Chagas disease.

Multimedia sobre historias de vida de profesionales de la Estomatología villaclareña / Multimedia on the life history of Dentistry professionals in Villa Clara

Fundamento: en la enseñanza de los hechos fundamentales de la historia de Cuba, tan importante como la cientificidad, es motivar al estudiante con la utilización de métodos que lo conviertan en un ente activo en la clase. Objetivo: diseñar una multimedia para el conocimiento de la historia de vida de profesionales de la Estomatología. Métodos: se realizó una investigación descriptiva con enfoque cualitativo en la que se utilizaron métodos teóricos: analítico-sintético, inductivo-deductivo, histórico-lógico y lógico-práctico; y como empíricos, el análisis documental: revisión de los programas Estomatología General Integral, III y IV e Historia de Cuba III, cuestionarios a estudiantes que reciben el programa y el criterio de especialistas para la valoración del producto. Resultados: se determinó que es posible insertar las historias de vida en la asignatura Historia de Cuba III, teniendo en cuenta las etapas cronológicas de la patria, en correspondencia con el accionar de estas personalidades, con lo cual coinciden los estudiantes, por lo que se diseñó una multimedia que contiene las historias de vida de algunos profesionales de la Estomatología en Villa Clara. Conclusiones: la multimedia diseñada constituye un medio de enseñanza propicio para el conocimiento del decursar de la Estomatología villaclareña, a partir de las historias de vida de los sujetos que participan en la formación de estos profesionales en el territorio. Fue valorada como pertinente, accesible, y como fuente histórica confiable por la veracidad de los datos que aporta(AU)

Multimedia sobre historias de vida de profesionales de la Estomatología villaclareña / Multimedia on the life history of Dentistry professionals in Villa Clara

Fundamento: en la enseñanza de los hechos fundamentales de la historia de Cuba, tan importante como la cientificidad, es motivar al estudiante con la utilización de métodos que lo conviertan en un ente activo en la clase. Objetivo: diseñar una multimedia para el conocimiento de la historia de vida de profesionales de la Estomatología. Métodos: se realizó una investigación descriptiva con enfoque cualitativo en la que se utilizaron métodos teóricos: analítico-sintético, inductivo-deductivo, histórico-lógico y lógico-práctico; y como empíricos, el análisis documental: revisión de los programas Estomatología General Integral, III y IV e Historia de Cuba III, cuestionarios a estudiantes que reciben el programa y el criterio de especialistas para la valoración del producto. Resultados: se determinó que es posible insertar las historias de vida en la asignatura Historia de Cuba III, teniendo en cuenta las etapas cronológicas de la patria, en correspondencia con el accionar de estas personalidades, con lo cual coinciden los estudiantes, por lo que se diseñó una multimedia que contiene las historias de vida de algunos profesionales de la Estomatología en Villa Clara. Conclusiones: la multimedia diseñada constituye un medio de enseñanza propicio para el conocimiento del decursar de la Estomatología villaclareña, a partir de las historias de vida de los sujetos que participan en la formación de estos profesionales en el territorio. Fue valorada como pertinente, accesible, y como fuente histórica confiable por la veracidad de los datos que aporta.

Background: in the teaching the basic facts of the history of Cuba, motivating the students with the use of methods that make them an active subject in the class is as important as the scientific approach. Objective: to design a multimedia for promoting the knowledge of the life history of Dentistry professionals. Methods: a descriptive study, with qualitative approach, was conducted. Theoretical methods were used: analytic-synthetic, inductive-deductive, historical-logical and logical-practical methods. The empirical methods included a documentary analysis (review of the syllabi of Comprehensive General Dentistry III and IV and History of Cuba III), questionnaires to students who study the subjects and expert judgment for valuation of the product. Results: it was found that it is possible to insert the life history of Dentistry professionals in the subject History of Cuba III, considering the chronological stages of the country, and in line with the actions of these personalities, something with which the students agree. Therefore, a multimedia containing the life history of some Dentistry professionals in Villa Clara was designed. Conclusions: the multimedia constitutes a proper teaching aid for promoting the knowledge about the history of Dentistry in Villa Clara, based on the life history of the individuals involved in the training of these professionals in the province. It was assessed as relevant, accessible, and as a reliable historical source because of the veracity of the data provided.

Comparative analyses of the ß-tubulin gene and molecular modeling reveal molecular insight into the colchicine resistance in kinetoplastids organisms.

Differential susceptibility to microtubule agents has been demonstrated between mammalian cells and kinetoplastid organisms such as Leishmania spp. and Trypanosoma spp. The aims of this study were to identify and characterize the architecture of the putative colchicine binding site of Leishmania spp. and investigate the molecular basis of colchicine resistance. We cloned and sequenced the ß-tubulin gene of Leishmania (Viannia) guyanensis and established the theoretical 3D model of the protein, using the crystallographic structure of the bovine protein as template. We identified mutations on the Leishmania ß-tubulin gene sequences on regions related to the putative colchicine-binding pocket, which generate amino acid substitutions and changes in the topology of this region, blocking the access of colchicine. The same mutations were found in the ß-tubulin sequence of kinetoplastid organisms such as Trypanosoma cruzi, T. brucei, and T. evansi. Using molecular modelling approaches, we demonstrated that conformational changes include an elongation and torsion of an α-helix structure and displacement to the inside of the pocket of one ß-sheet that hinders access of colchicine. We propose that kinetoplastid organisms show resistance to colchicine due to amino acids substitutions that generate structural changes in the putative colchicine-binding domain, which prevent colchicine access.

A PCR assay for the identification of Leishmania species of the Viannia subgenus / Un ensayo de PCR para identificar especies de Leishmania del subgénero Viannia

We have identified a novel DNA sequence of 500 bp (β500-DNA) on the Leishmania (Viannia) subgenus, located in the intergenic region of one of the loci of the β-tubulin gene family. The sequence analysis showed that this sequence has no homology to any other sequence described so far, including the β-tubulin gene. We improved a specific β500-PCR assay, which generated a PCR product of 375 bp for total genomic DNA from Leishmania strains belonging to the L. (Viannia) subgenus. In contrast, no amplification was found when using genomic DNA from species of L. (Leishmania) subgenus or other organisms. Under our PCR conditions, the lower detection limit was 1 fg when a purified DNA clone (pLgβ4), which contains one copy of the β500-DNA sequence, was used. The β500-DNA PCR assay confirmed the preliminary diagnosis of cutaneous leishmaniasis in clinical samples in which the Montenegro skin test was positive and parasite cultures were negative. The analytical specificity and the sensitivity of the PCR assay provide a tool for epidemiological studies of the disease.

En este trabajo identificamos una nueva secuencia de DNA de 500 pb (β500-DNA) en Leishmania del subgénero Leishmania (Viannia), localizada en la región intergénica de uno de los loci de la familia de los genes de la β tubulina. El análisis de secuencia mostró que β500 no tiene homología con ninguna otra secuencia previamente descrita, incluido el gen de la β tubulina. Nosotros implementamos un ensayo de PCR específico para β500, β500-PCR, que genera un producto de PCR de 375 pb a partir del DNA genómico de cepas de Leishmania pertenecientes al subgénero L. (Viannia). No hubo amplificación alguna cuando se utilizó el DNA genómico de especies del subgénero L. (Leishmania) o el de otros organismos. En las condiciones establecidas, utilizando DNA purificado del clon pLgβ4, que contiene una copia de la secuencia de DNA de β500, el límite de detección más bajo fue de 1 fentogramo. El ensayo β500-PCR confirmó el diagnóstico preliminar de leishmaniasis cutánea en muestras clínicas de pacientes positivas a la prueba de Montenegro y negativas en el cultivo de parásitos. La especificidad y sensibilidad analítica del ensayo de PCR proporciona una herramienta para estudios epidemiológicos de la enfermedad.

The 6-phosphogluconate dehydrogenase of Leishmania (Leishmania) mexicana: gene characterization and protein structure prediction.

6-Phosphogluconate dehydrogenase (6PGDH) is a key enzyme of the oxidative branch involved in the generation of NADPH and ribulose 5-phosphate. In the present work, we describe the cloning, sequencing and characterization of a 6PGDH gene from Leishmania (Leishmania) mexicana. The gene encodes a polypeptide chain of 479 amino acid residues with a predicted molecular mass of 52 kDa and a pI of 5.77. The recombinant protein possesses a dimeric quaternary structure and displays kinetic parameter values intermediate between those reported for Trypanosoma brucei and T. cruzi with apparent K(m) values of 6.93 and 5.2 µM for 6PG and NADP(+), respectively. The three-dimensional structure of the enzymes of Leishmania and T. cruzi were modelled from their amino acid sequence using the crystal structure of the enzyme of T. brucei as template. The amino acid residues located in the 6PGDH C-terminal region, which are known to participate in the salt bridges maintaining the protein dimeric structure, differed significantly among the enzymes of Leishmania, T. cruzi, and T. brucei. Our results strongly suggest that 6PGDH can be selected as a potential target for the development of new therapeutic drugs in order to improve existing chemotherapeutic treatments against these parasites.

Nuevo espectro de tuberculide papulosa: aspectos clínicos e histopatológicos a propósito de un caso / New spectrum of papular tuberculid: clinical and pathological aspects due to a case

Presentamos el caso de un paciente varón de 59 años de edad quien inicialmente presentó un cuadro clínico de tuberculide papulosa en rostro o tuberculide rosaceiforme, que progresivamente se extiende a cuero cabelludo, cuello, tronco y miembros superiores. Se plantea la posibilidad de que este nuevo espectro de tuberculides constituiría formas de tuberculosis cutánea paucibacilar.

We present the case of a 59-years-old male patient who was diagnosed of papular tuberculid or rosaceiform tuberculid in face, with progressive extension to the scalp, neck, trunk and arms. We propose that this new spectrum of tuberculids could be a paucibacilar form of cutaneous tuberculosis.

Amiodarone and miltefosine act synergistically against Leishmania mexicana and can induce parasitological cure in a murine model of cutaneous leishmaniasis.

Leishmaniasis is parasitic disease that is an important problem of public health worldwide. Intramuscularly administered glucantime and pentostam are the most common drugs used for treatment of this disease, but they have significant limitations due to toxicity and increasing resistance. A recent breakthrough has been the introduction of orally administered miltefosine for the treatment of visceral, cutaneous, and mucocutaneous leishmaniasis, but the relative high cost and concerns about teratogenicity have limited the use of this drug. Searching for alternative drugs, we previously demonstrated that the antiarrhythmic drug amiodarone is active against Leishmania mexicana promastigotes and intracellular amastigotes, acting via disruption of intracellular Ca(2+) homeostasis (specifically at the mitochondrion and the acidocalcisomes of these parasites) and through inhibition of the parasite's de novo sterol biosynthesis (X. Serrano-Martín, Y. García-Marchan, A. Fernandez, N. Rodriguez, H. Rojas, G. Visbal, and G. Benaim, Antimicrob. Agents Chemother. 53:1403-1410, 2009). In the present work, we found that miltefosine also disrupts the parasite's intracellular Ca(2+) homeostasis, in this case by inducing a large increase in intracellular Ca(2+) levels, probably through the activation of a plasma membrane Ca(2+) channel. We also investigated the in vitro and in vivo activities of amiodarone and miltefosine, used alone or in combination, on L. mexicana. It was found that the drug combination had synergistic effects on the proliferation of intracellular amastigotes growing inside macrophages and led 90% of parasitological cures in a murine model of leishmaniasis, as revealed by a PCR assay using a novel DNA sequence specific for L. mexicana.

Identificación de una secuencia de ADN genómico de Leishmania especifica del subgénero Leishmania / Identification of a genomic DNA sequence of Leishmania, specific of Leishmania subgenus

Leishmania es el agente causante de la compleja enfermedad conocida como leishmaniasis. Las distintas especies de este parásito protozoario se encuentran agrupadas en dos subgéneros, Viannia y Leishmania, de acuerdo a su desarrollo en el mosquito vector. Un ensayo de PCR, β500-PCR, específico del subgénero Viannia, ha sido desarrollado utilizando la secuencia de ADN genómico denominada β500. En este trabajo se presenta el aislamiento e identificación de una secuencia genómica de 280 pb, L280, a partir del ADN genómico de Leishmania (Leishmania) mexicana luego de aplicar el ensayo β500-PCR en condiciones de baja rigurosidad. La secuenciación parcial de L280 permitió diseñar un ensayo de PCR (L280-PCR) que generó un producto de amplificación de 260 pb, en distintas condiciones de rigurosidad, cuando se utilizó el ADN genómico de distintas especies pertenecientes al subgénero Leishmania. El ensayo L280-PCR resultó negativo para el ADN genómico de distintas especies del subgénero Viannia al igual que para el ADN de otros organismos kinetoplastidos o humano. Los resultados sugieren que el ensayo L280-PCR es específico del subgénero Leishmania.

Leishmania is the causal agent of the leishmaniasis disease. The different species of this protozoa parasite are grouped in two subgenera, Viannia and Leishmania, according to their development in the sandfly vector. A specific PCR assay, β500-PCR, has been developed for the Viannia subgenus using the genomic β500 DNA sequence. In the present work we present the isolation and identification of a genomic sequence of 280 bp, L280, obtained from genomic DNA of Leishmania (Leishmania) mexicana after application of the β500-PCR assay at low stringency. After partial sequencing of L280 a PCR assay was generated, L280-PCR, this yielded a product of 260 bp at different conditions of stringency, when genomic DNA of different species of Leishmania subgenus was used. The L280-PCR assay was negative to genomic DNA of species belonging to the Viannia subgenus and also to other kinetoplastid organisms and human. The results suggest specificity of the L280-PCR assay for the Leishmania subgenus.

Glibenclamide, a blocker of K+(ATP) channels, shows antileishmanial activity in experimental murine cutaneous leishmaniasis.

Glibenclamide reduced the rate of lesion growth in BALB/c mice infected with Leishmania (Leishmania) mexicana, the effect was dose dependent, and the highest dose proved more effective than glucantime. Cross-resistance to glucantime was found in animals infected with a glibenclamide-resistant line, but combined therapy reduced lesion progression even in the glibenclamide-resistant strain.

Detección parsitológica y molecular de infecciones naturales por trypanosoma evansy y trypanosoma vivax en búfalos de agua (bubalus bubalis) y chigüires (hydrochoerus hydrochaeris) en los estados Apure Cojedes y Guárico Venezuela / Parasitological and molecular detection of natural infection by trypanosoma evansi and trypanosoma vivax in water buffaloes (bubalus bubalis) and capybaras (hydrochoerus hydrochaeris) in Apure Cojedes and Guarico states Venezuela

Este estudio evaluó la presencia de Trypanosoma evansi y Trypanosoma vivax en búfalos de agua (Bubalus bubalis) y chigüires (Hydrochoerus hydrochaeris) de tres estados de Venezuela por técnicas parasitológicas y moleculares (frotis teñido: FT; microcentrifugación capilar: TMC; reacción en cadena de la Polimerasa: PCR), estableciéndose el porcentaje de infecciones activas. En 316 muestras sanguíneas de búfalos las tasas de detección para FT, TMC y PCR fueron de 20 (6,33 por ciento), 36 (11,39 por ciento) y 60 (18,98 por ciento),respectivamente. Por PCR se caracterizó a T. vivax como la especie responsable de todas las infecciones, no detectándose presencia de T. evansi. En 186 muestras de chigüires FT y TMC identificaron positividad en 36 (19,35 por ciento) y 71 (38,17 por ciento) de estas, respectivamente, con altas parasitemias; en 27 de estas se observaron tripanosomas del Subgénero Megatrypanum. Por PCR se caracterizaron como T. evansi 51 muestras de chigüires, seis con resultados TMC-negativo. No se detectó T. vivax en chigüires. Un análisis de t-student estableció diferencias (p<0.05) entre los valores de hematocrito (Ht) de búfalos positivos y negativos a tripanosomas; mientras que por análisis de varianza se detectó efecto (p< 0.05) del grado de parasitemia sobre el Ht. No hubo ningún efecto (p>0.05) al realizar los mismos análisis para las muestras de chigüires.

Detección diferencial de trypanosoma evansi y trypanosoma vivax mediante un ensayo de reacción en cadena de la polimerasa / Differential detection of trypanosoma evansi and trypanosoma vivax through a polymerase chain reaction assay

El propósito de este trabajo fue optimizar un ensayo de reacción en cadena de la polimerasa (PCR) para efectuar el diagnóstico diferencial de Trypanosoma evansi y Trypanosoma vivax, usando cebadores previamente descritos: 21-mer/22-mer e ILO1264/ILO1265. La especificidad se evaluó efectuando pruebas preliminares sobre muestras de ADN purificado de diversas especies de parásitos y sobre mezclas de éstas. La sensibilidad se valoró empleando diluciones de ADN de aislados de referencia de T. evansi y T. vivax y concentraciones variables de cebadores. El ensayo optimizado mostró una sensibilidad de 10 pg de ADN, con especificidad para el Subgénero Trypanozoon (cebadores 21-mer/22-mer) y especificidad absoluta para T. vivax (cebadores ILO1264/ILO1265). Ambos grupos de cebadores mostraron potencialidad para detectar infecciones mixtas T. evansi/T. vivax. Un análisis de hibridación sobre el patrón de cariotipo de diversos Kinetoplastida, usando la sonda Te-ADN, mostró su carácter repetitivo y la distribución de su secuencia en diferentes cromosomas de T. evansi TE0. La PCR fue evaluada como herramienta diagnóstico de la presencia de tripanosomas en muestras sanguíneas de animales con infecciones experimentales, demostrando alta sensibilidad al detectar positividad hasta 72 horas antes que lo hicieran los métodos parasitológicos. Este ensayo también fue evaluado sobre muestras sanguíneas de búfalos de agua con infecciones naturales, mostrando alta sensibilidad y especificidad al detectar 9/86 muestras como positivas a T. vivax, revelando una tasa de infección activa de 10,47 por ciento; valor tres veces superior a los resultados de la evaluación microscópica de frotis teñidos (3,49 por ciento) y casi dos veces superior (6,98 por ciento) a la técnica parasitológica de microcentrifugación capilar. En ninguna de las muestras de búfalos evaluadas se detectó T. evansi.