Targeted regulation of protein levels is an important tool to investigate the role of proteins essential for cell function and development. In recent years, methods based on the Escherichia coli dihydrofolate reductase destabilization domain (ecDHFR DD) have been established and used in various cell types. ecDHFR DD destabilizes the fused protein of interest and causes its degradation by protea...

We have infected BALB/c and C57BL/6 mice with a cloned Leishmania mexicana amazonensis population, obtained from the "Maria" strain. Progression of infection and histopathological examination confirmed the extreme susceptibility of BALB/c mice and the resistant pattern of the C57 BL/6. Anti-Leishmania antibody titers were higher in BALB/c than in C57BL/6 mice through the period of infection. Te...

A total of 519 wild animals belonging to eleven species were collected during a two year study in a cutaneous leishmaniasis endemic area in Venezuela (La Matica, Lara State). The animals were captured in home-made Tomahawk-like traps baited with maize, bananas or other available local fruits, and parasites were isolated from 27 specimens. Two different species were found naturally infected with...

A flow cytometric technique was developed for detection of amastigotes of the protozoan Leishmania infantum in human nonadherent monocyte-derived macrophages. The cells were fixed and permeabilized with paraformaldehyde-ethanol, and intracellular amastigotes were labeled with Leishmania lipophosphoglycan-specific monoclonal antibody. Results showed that flow cytometry provided accurate quantifi...

Functional analysis of genes from parasitic helminths requires, at the present time, heterologous expression. We have adapted the well-characterized system of transfection in Leishmania protozoal parasites, as a means of analysing the effect of single filarial genes on the mammalian immune system. For example, testing the function of the Brugia malayi abundant larval transcript (ALT) gene-trans...

The ultrastructural study in different tissues of mice experimentally infected with isolates of Trypanosoma evansi, Trypanosoma cruzi, and Leishmania mexicana reveals changes in cardiac myocytes, skeletal muscle fibers, and hepatic, adrenal, kidney, and spleen cells. Some of these changes were cytoarchitectural and others consisted of necrosis. Alterations in the microvasculature were also foun...

The protozoan Leishmania mexicana parasite causes chronic non-healing cutaneous lesions in humans and mice with poor parasite control. The mechanisms preventing the development of a protective immune response against this parasite are unclear. Here we provide data demonstrating that parasite sequestration by neutrophils is responsible for disease progression in mice. Within hours of infection L...

Immunization with the GP46/M-2 membrane glycoprotein of Leishmania amazonensis has been shown to induce a protective immune response against infection. We have surveyed a variety of trypanosomatid species and genera for the presence and expression of this gene family, information that will be relevant to future vaccine studies against leishmaniasis. Molecular karyotype analysis revealed the pre...

The effect of ergosterol depletion by ketoconazole on the leishmanicidal activity of the pore-forming antibiotic amphotericin B (AmB) was investigated. Leishmania mexicana promastigotes were lysed within minutes by the addition of micromolar concentrations of AmB (0.5 microM) but became insensitive to AmB after growth in the presence of ketoconazole (0.25 microM, 90 h). Lipid chromatographic an...

We present the genome sequence of Leishmania mexicana MNYC/BZ/62/M379 modified to express Cas9 and T7 RNA-polymerase, revealing high similarity reference (MHOM/GT2001/U1103). Through RNAseq-based annotation coding sequences untranslated regions, we provide primer for construct sgRNA template generation CRISPR-assisted gene deletion endogenous tagging.