Susceptibilidad in vitro a infección por Leishmania y sensibilidad a medicamentos difiere según tipo de macrófagos

RESUMEN Introducción: Los sistemas in vitro son útiles en la evaluación de compuestos con actividad biológica, permitiendo por ejemplo, determinar el potencial citotóxico y leishmanicida de un compuesto.Objetivo: Identificar el tipo de célula mamífera que permita una óptima infección in vitro por Leishmania y que constituya el sistema adecuado para el tamizaje in vitro de compuestos con actividad anti-Leishmania.Métodos: La susceptibilidad de las células a infección por L. panamensis se evalúo según la Concentración Infectiva 50 determinada por microscopía de luz y citometría de flujo; la supervivencia intracelular de los amastigotes se evaluó por microscopía de fluorescencia y la sensibilidad de las células a anfotericina B y antimoniato de meglumina se evalúo por espectrofotometría.Resultados: Los cultivos primarios son más susceptibles a la infección por L. panamensis in vitro. Sin embargo, la supervivencia intracelular del parásito fue mejor en U-937. Por su parte, la sensibilidad de las células a anfotericina B y antimoniato de meglumina vario según el tipo de célula.Conclusiones: Las células U-937 son las adecuadas para la infección por Leishmania porque: a) presentan crecimiento ilimitado y se les puede inducir transformación a macrófagos. b) la susceptibilidad a la infección por Leishmania es similar a la observada en cultivos primarios de macrófagos y c) permiten mayor supervivencia de los amastigotes luego de la infección. Adicionalmente, las células U-937 son menos sensibles a la acción de los fármacos comúnmente utilizados como control en la detección de compuestos con actividad leishmanicida. Salud UIS 2010; 42:   Palabras clave: Susceptibilidad a infección por Leishmania, células U-937, células THP-1, células Vero, células J774A,1, macrófagos ABSTRACT Introduction: In vitro systems are useful in the evaluation of compounds with biological activity determining the cytotoxic and leishmanicidal activity of the candidates.Objective: To identify the mammalian cell that allows the optimal in vitro infection by Leishmania and therefore, identify the suitable system for the in vitro evaluation of leishmanicidal activity of drugs.Methodology: The susceptibility to the infection by L. panamensis was evaluated according to the Infective Concentration 50 tested by light microscopy and flow citometry; the intracellular survival of amastigotes was determined by fluorescence microscopy and the sensitivity to amphotericine B and meglumine  antimoniate was evaluated by spectrophotometry.Results: The primary culture cells were more susceptible to the in vitro infection by L. panamensis because they did require fewer parasites per cell ratio to achieve the 50% infection rate whereas the intracellular survival of parasites was better in the U-937 cells. All cells showed differential sensitivity to amphotericine B and meglumine antimoniate.Conclusion: The U-937 cells are the most suitable model for the in vitro infection by L. panamensis because: a) they are a cell line with unlimited growth where transformation into macrophages can be induced. b) The susceptibility to infection by L. panamensis is similar to that observed in primary cultures macrophages and, c) They allow the intracellular survival of amastigotes after the infection process. In addition the U-937 cells are less sensitive to the action of the commonly drugs used as a control in the screening of compounds with leishmanicidal activity. Salud UIS 2010; 42:   Keywords: Susceptibility to infection by Leishmania, U-937 cells, THP-1 cells, Vero cells, J774A.1 cells, macrophages

Susceptibilidad in vitro a infección por Leishmania y sensibilidad a medicamentos difiere según tipo de macrófagos

RESUMEN Introducción: Los sistemas in vitro son útiles en la evaluación de compuestos con actividad biológica, permitiendo por ejemplo, determinar el potencial citotóxico y leishmanicida de un compuesto.Objetivo: Identificar el tipo de célula mamífera que permita una óptima infección in vitro por Leishmania y que constituya el sistema adecuado para el tamizaje in vitro de compuestos con actividad anti-Leishmania.Métodos: La susceptibilidad de las células a infección por L. panamensis se evalúo según la Concentración Infectiva 50 determinada por microscopía de luz y citometría de flujo; la supervivencia intracelular de los amastigotes se evaluó por microscopía de fluorescencia y la sensibilidad de las células a anfotericina B y antimoniato de meglumina se evalúo por espectrofotometría.Resultados: Los cultivos primarios son más susceptibles a la infección por L. panamensis in vitro. Sin embargo, la supervivencia intracelular del parásito fue mejor en U-937. Por su parte, la sensibilidad de las células a anfotericina B y antimoniato de meglumina vario según el tipo de célula.Conclusiones: Las células U-937 son las adecuadas para la infección por Leishmania porque: a) presentan crecimiento ilimitado y se les puede inducir transformación a macrófagos. b) la susceptibilidad a la infección por Leishmania es similar a la observada en cultivos primarios de macrófagos y c) permiten mayor supervivencia de los amastigotes luego de la infección. Adicionalmente, las células U-937 son menos sensibles a la acción de los fármacos comúnmente utilizados como control en la detección de compuestos con actividad leishmanicida. Salud UIS 2010; 42:   Palabras clave: Susceptibilidad a infección por Leishmania, células U-937, células THP-1, células Vero, células J774A,1, macrófagos ABSTRACT Introduction: In vitro systems are useful in the evaluation of compounds with biological activity determining the cytotoxic and leishmanicidal activity of the candidates.Objective: To identify the mammalian cell that allows the optimal in vitro infection by Leishmania and therefore, identify the suitable system for the in vitro evaluation of leishmanicidal activity of drugs.Methodology: The susceptibility to the infection by L. panamensis was evaluated according to the Infective Concentration 50 tested by light microscopy and flow citometry; the intracellular survival of amastigotes was determined by fluorescence microscopy and the sensitivity to amphotericine B and meglumine  antimoniate was evaluated by spectrophotometry.Results: The primary culture cells were more susceptible to the in vitro infection by L. panamensis because they did require fewer parasites per cell ratio to achieve the 50% infection rate whereas the intracellular survival of parasites was better in the U-937 cells. All cells showed differential sensitivity to amphotericine B and meglumine antimoniate.Conclusion: The U-937 cells are the most suitable model for the in vitro infection by L. panamensis because: a) they are a cell line with unlimited growth where transformation into macrophages can be induced. b) The susceptibility to infection by L. panamensis is similar to that observed in primary cultures macrophages and, c) They allow the intracellular survival of amastigotes after the infection process. In addition the U-937 cells are less sensitive to the action of the commonly drugs used as a control in the screening of compounds with leishmanicidal activity. Salud UIS 2010; 42:   Keywords: Susceptibility to infection by Leishmania, U-937 cells, THP-1 cells, Vero cells, J774A.1 cells, macrophages

Estructura y actividad de sapogeninas triterpenicas

La leishmaniasis es una enfermedad ampliamente difundida en América latina, desafortunadamente para su tratamiento hay muy pocas drogas, que tambien tienen una baja efectividad. Por esta razón el mundo urge de nuevas y más efectivas moléculas. En la búsqueda de sustancias antiparasitarias se detecto una alta actividad leishmanicida de Sapindus saponaria ricas en sapogeninas. El fraccionamiento y posterior purificación se obtuvieron tres sapogeninas triterpenicas cuyas estructuras se describen ene este articulo

Copy Number Variants Are Ovarian Cancer Risk Alleles at Known and Novel Risk Loci

BACKGROUND: Known risk alleles for epithelial ovarian cancer (EOC) account for approximately 40% of the heritability for EOC. Copy number variants (CNVs) have not been investigated as EOC risk alleles in a large population cohort. METHODS: Single nucleotide polymorphism array data from 13 071 EOC cases and 17 306 controls of White European ancestry were used to identify CNVs associated with EOC risk using a rare admixture maximum likelihood test for gene burden and a by-probe ratio test. We performed enrichment analysis of CNVs at known EOC risk loci and functional biofeatures in ovarian cancer-related cell types. RESULTS: We identified statistically significant risk associations with CNVs at known EOC risk genes; BRCA1 (PEOC = 1.60E-21; OREOC = 8.24), RAD51C (Phigh-grade serous ovarian cancer [HGSOC] = 5.5E-4; odds ratio [OR]HGSOC = 5.74 del), and BRCA2 (PHGSOC = 7.0E-4; ORHGSOC = 3.31 deletion). Four suggestive associations (P < .001) were identified for rare CNVs. Risk-associated CNVs were enriched (P < .05) at known EOC risk loci identified by genome-wide association study. Noncoding CNVs were enriched in active promoters and insulators in EOC-related cell types. CONCLUSIONS: CNVs in BRCA1 have been previously reported in smaller studies, but their observed frequency in this large population-based cohort, along with the CNVs observed at BRCA2 and RAD51C gene loci in EOC cases, suggests that these CNVs are potentially pathogenic and may contribute to the spectrum of disease-causing mutations in these genes. CNVs are likely to occur in a wider set of susceptibility regions, with potential implications for clinical genetic testing and disease prevention

Genetic Data from Nearly 63,000 Women of European Descent Predicts DNA Methylation Biomarkers and Epithelial Ovarian Cancer Risk

DNA methylation is instrumental for gene regulation. Global changes in the epigenetic landscape have been recognized as a hallmark of cancer. However, the role of DNA methylation in epithelial ovarian cancer (EOC) remains unclear. In this study, high-density genetic and DNA methylation data in white blood cells from the Framingham Heart Study (N = 1,595) were used to build genetic models to predict DNA methylation levels. These prediction models were then applied to the summary statistics of a genome-wide association study (GWAS) of ovarian cancer including 22,406 EOC cases and 40,941 controls to investigate genetically predicted DNA methylation levels in association with EOC risk. Among 62,938 CpG sites investigated, genetically predicted methylation levels at 89 CpG were significantly associated with EOC risk at a Bonferroni-corrected threshold of P <7.94 x 10(-7). Of them, 87 were located at GWAS-identified EOC susceptibility regions and two resided in a genomic region not previously reported to be associated with EOC risk. Integrative analyses of genetic, methylation, and gene expression data identified consistent directions of associations across 12 CpG, five genes, and EOC risk, suggesting that methylation at these 12 CpG may influence EOC risk by regulating expression of these five genes, namely MAPT, HOXB3, ABHD8, ARHGAP27, and SKAP1. We identified novel DNA methylation markers associated with EOC risk and propose that methylation at multiple CpG may affect EOC risk via regulation of gene expression. Significance: Identification of novel DNA methylation markers associated with EOC risk suggests that methylation at multiple CpG may affect EOC risk through regulation of gene expression.Peer reviewe

Shared heritability and functional enrichment across six solid cancers

Correction: Nature Communications 10 (2019): art. 4386 DOI: 10.1038/s41467-019-12095-8Quantifying the genetic correlation between cancers can provide important insights into the mechanisms driving cancer etiology. Using genome-wide association study summary statistics across six cancer types based on a total of 296,215 cases and 301,319 controls of European ancestry, here we estimate the pair-wise genetic correlations between breast, colorectal, head/neck, lung, ovary and prostate cancer, and between cancers and 38 other diseases. We observed statistically significant genetic correlations between lung and head/neck cancer (r(g) = 0.57, p = 4.6 x 10(-8)), breast and ovarian cancer (r(g) = 0.24, p = 7 x 10(-5)), breast and lung cancer (r(g) = 0.18, p = 1.5 x 10(-6)) and breast and colorectal cancer (r(g) = 0.15, p = 1.1 x 10(-4)). We also found that multiple cancers are genetically correlated with non-cancer traits including smoking, psychiatric diseases and metabolic characteristics. Functional enrichment analysis revealed a significant excess contribution of conserved and regulatory regions to cancer heritability. Our comprehensive analysis of cross-cancer heritability suggests that solid tumors arising across tissues share in part a common germline genetic basis.Peer reviewe

In Vitro and In Vivo Efficacy of Ether Lipid Edelfosine against Leishmania spp. and SbV-Resistant Parasites

Leishmaniasis represents a major international health problem, has a high morbidity and mortality rate, and is classified as an emerging and uncontrolled disease by the World Health Organization. The migration of population from endemic to nonendemic areas, and tourist activities in endemic regions are spreading the disease to new areas. Unfortunately, treatment of leishmaniasis is far from satisfactory, with only a few drugs available that show significant side-effects. Here, we show in vitro and in vivo evidence for the antileishmanial activity of the ether phospholipid edelfosine, being effective against a wide number of Leishmania spp. causing cutaneous, mucocutaneous and visceral leishmaniasis. Our experimental mouse and hamster models demonstrated not only a significant antileishmanial activity of edelfosine oral administration against different wild-type Leishmania spp., but also against parasites resistant to pentavalent antimonials, which constitute the first line of treatment worldwide. In addition, edelfosine exerted a higher antileishmanial activity and a lower proneness to generate drug resistance than miltefosine, the first drug against leishmaniasis that can be administered orally. These data, together with our previous findings, showing an anti-inflammatory action and a very low toxicity profile, suggest that edelfosine is a promising orally administered drug for leishmaniasis, thus warranting clinical evaluation