ABSTRACT
The municipality of Bauru, São Paulo, Brazil, is an area endemic for leishmaniasis. At the zoo, a spider monkey (Ateles paniscus) showed nonpathognomonic symptoms, such as weight loss and pale mucous membranes. Blood was collected from the jugular vein and investigated for the presence of Leishmania spp. DNA by polymerase chain reaction and restriction fragment length polymorphism. Parasite DNA was detected, and the pattern observed was identical to Leishmania amazonensis. This study presents molecular evidence of L. amazonensis infection in a captive spider monkey.
Subject(s)
Leishmania/classification , Leishmania/isolation & purification , Leishmaniasis/veterinary , Monkey Diseases/parasitology , Animals , Animals, Zoo , Atelinae , Brazil/epidemiology , DNA, Protozoan/genetics , Leishmaniasis/epidemiology , Leishmaniasis/parasitology , Male , Monkey Diseases/epidemiology , Real-Time Polymerase Chain Reaction/veterinaryABSTRACT
The protozoan Trypanosoma cruzi is the causative agent of the neglected infectious illness Chagas disease. During its life cycle it differentiates into replicative and non-replicative life stages. So far, T. cruzi cell division has been investigated by transcriptomics but not by proteomics approaches. Here we show the first quantitative proteome analysis of T. cruzi cell division. T. cruzi epimastigote cultures were subject to synchronization with hydroxyurea and harvested at different time points. Analysis by flow cytometry, bright field and fluorescence microscopy indicated that samples collected at 0 h, 2 h, 6 h and 14 h overrepresented G1, G1-S, S and M cell cycle phases, respectively. After trypsin digestion of these samples, the resulting peptides were labelled with iTRAQ and subjected to LC-MS/MS. Also, iTRAQ-labelled phosphopeptides were enriched with TiO2 to access the phosphoproteome. Overall, 597 protein groups and 94 phosphopeptides presented regulation with the most remarkable variation in abundance at 6 h (S-phase). Comparison of our proteomic data to previous transcriptome-wise analysis of epimastigote cell cycle showed 16 sequence entries in common, with the highest mRNA/protein correlation observed in transcripts with peak abundance in G1-phase. Our data revealed regulated proteins and phosphopeptides which play important roles in the control of cell division in other organisms and some of them were previously detected in the nucleus or associated with T. cruzi chromatin.
Subject(s)
Cell Cycle , Phosphoproteins/metabolism , Proteomics/methods , Protozoan Proteins/metabolism , Trypanosoma cruzi/metabolism , Cell Nucleus/metabolism , Chromatin/metabolism , Chromatography, Liquid/methods , Flow Cytometry , Microscopy, Fluorescence , Tandem Mass Spectrometry/methods , Transcriptome , Trypanosoma cruzi/cytologyABSTRACT
A leishmaniose visceral (LV) é uma zoonose endêmica na América Latina e 96% dos casos de LV são diagnosticados no Brasil. A coinfecção HIV-LV tem sido diagnosticada em áreas endêmicas e não endêmicas para LV. O aumento do número de casos de coinfecções em todo o mundo deve-se, em parte, à coincidência das áreas de circulação desses organismos. Deve-se ressaltar que a concomitância das duas infecções é potencialmente deletéria, portanto a associação dos dois patógenos constitui um desafio para o diagnóstico e controle da LV. A interação entre Leishmania e HIV é prejudicial, pois há o risco de progressão rápida de ambas as doenças por compartilharem mecanismos imunológicos semelhantes. Neste relato, é apresentado o caso de um paciente com infecção pelo HIV associada à LV, que evoluiu rapidamente para o óbito.
Subject(s)
Leishmaniasis, Visceral , Acquired Immunodeficiency Syndrome , HIV , CoinfectionABSTRACT
Mutantes auxotróficos e Nif de Bacillus azotofixans foram isolados a partir de culturas tratadas com EMS, empregando-se um método de enriquecimento com penicilina, desenvolvido para este microrganismo. A frequência de obtençäo de mutantes auxotróficos (0,19 por cento) foi cerce de três vezes maior do que a de obtençäo de mutantes Nif(0,063 por cento), embora ambas as frequências fossem extremamente baixas. Os fatores de crescimento exigidos pelos mutantes auxotróficos incluiam aminoácidos (47,05 por cento), bases nitrogenadas (17,64 por cento) ou vitaminas (11,76 por cento). Estes mutantes seräo posteriormente usados em experimentos de transferência gênica, realizados com o objetivo de se localizar os genes nif no cromossomo desse microrganismo