185 research outputs found
Reaching for the Holy Grail: insights from infection/cure models on the prospects for vaccines for Trypanosoma cruzi infection
Prevention of Trypanosoma cruzi infection in mammals likely
depends on either prevention of the invading trypomastigotes
from infecting host cells or the rapid recognition and killing
of the newly infected cells by T. cruzi-specific T cells. We
show here that multiple rounds of infection and cure (by drug
therapy) fails to protect mice from reinfection, despite the
generation of potent T cell responses. This disappointing result
is similar to that obtained with many other vaccine protocols
used in attempts to protect animals from T. cruzi infection. We
have previously shown that immune recognition of T. cruzi
infection is significantly delayed both at the systemic level
and at the level of the infected host cell. The systemic delay
appears to be the result of a stealth infection process that
fails to trigger substantial innate recognition mechanisms while
the delay at the cellular level is related to the
immunodominance of highly variable gene family proteins, in
particular those of the trans-sialidase family. Here we discuss
how these previous studies and the new findings herein impact
our thoughts on the potential of prophylactic vaccination to
serve a productive role in the prevention of T. cruzi infection
and Chagas disease
Chromosome level assembly of the hybrid Trypanosoma cruzi genome
<p>Abstract</p> <p>Background</p> <p>In contrast to the essentially fully assembled genome sequences of the kinetoplastid pathogens <it>Leishmania major </it>and <it>Trypanosoma brucei </it>the assembly of the <it>Trypanosoma cruzi </it>genome has been hindered by its repetitive nature and the fact that the reference strain (CL Brener) is a hybrid of two distinct lineages. In this work, the majority of the contigs and scaffolds were assembled into pairs of homologous chromosomes based on predicted parental haplotype, inference from TriTryp synteny maps and the use of end sequences from <it>T. cruzi </it>BAC libraries.</p> <p>Results</p> <p>Ultimately, 41 pairs of chromosomes were assembled using this approach, a number in agreement with the predicted number of <it>T. cruzi </it>chromosomes based upon pulse field gel analysis, with over 90% (21133 of 23216) of the genes annotated in the genome represented. The approach was substantiated through the use of Southern blot analysis to confirm the mapping of BAC clones using as probes the genes they are predicted to contain, and each chromosome construction was visually validated to ensure sufficient evidence was present to support the organization. While many members of large gene families are incorporated into the chromosome assemblies, the majority of genes excluded from the chromosomes belong to gene families, as these genes are frequently impossible to accurately position.</p> <p>Conclusion</p> <p>Now assembled, these chromosomes bring <it>T. cruzi </it>to the same level of organization as its kinetoplastid relatives and have been used as the basis for the <it>T. cruzi </it>genome in TriTrypDB, a trypanosome database of EuPathDB. In addition, they will provide the foundation for analyses such as reverse genetics, where the location of genes and their alleles and/or paralogues is necessary and comparative genome hybridization analyses (CGH), where a chromosome-level view of the genome is ideal.</p
From Questions to Effective Answers: On the Utility of Knowledge-Driven Querying Systems for Life Sciences Data
We compare two distinct approaches for querying data in the context of the
life sciences. The first approach utilizes conventional databases to store the
data and intuitive form-based interfaces to facilitate easy querying of the
data. These interfaces could be seen as implementing a set of "pre-canned"
queries commonly used by the life science researchers that we study. The second
approach is based on semantic Web technologies and is knowledge (model) driven.
It utilizes a large OWL ontology and same datasets as before but associated as
RDF instances of the ontology concepts. An intuitive interface is provided that
allows the formulation of RDF triples-based queries. Both these approaches are
being used in parallel by a team of cell biologists in their daily research
activities, with the objective of gradually replacing the conventional approach
with the knowledge-driven one. This provides us with a valuable opportunity to
compare and qualitatively evaluate the two approaches. We describe several
benefits of the knowledge-driven approach in comparison to the traditional way
of accessing data, and highlight a few limitations as well. We believe that our
analysis not only explicitly highlights the specific benefits and limitations
of semantic Web technologies in our context but also contributes toward
effective ways of translating a question in a researcher's mind into precise
computational queries with the intent of obtaining effective answers from the
data. While researchers often assume the benefits of semantic Web technologies,
we explicitly illustrate these in practice
The covertisation of norms in contact situations : The influence of the nonnative speaker on native speaker behaviour
Recombination events. a) Detailed information of 2087 recombination events detected in TcTS gene family. b) Summary of number and percentage of newly annotated TcTS and previously annotated TcTS participating in recombination events as recombinant product, major donor or minor donor. (XLSX 549 kb
Evaluation of high efficiency gene knockout strategies for Trypanosoma cruzi
<p>Abstract</p> <p>Background</p> <p><it>Trypanosoma cruzi</it>, a kinetoplastid protozoan parasite that causes Chagas disease, infects approximately 15 million people in Central and South America. In contrast to the substantial <it>in silico </it>studies of the <it>T. cruzi </it>genome, transcriptome, and proteome, only a few genes have been experimentally characterized and validated, mainly due to the lack of facile methods for gene manipulation needed for reverse genetic studies. Current strategies for gene disruption in <it>T. cruzi </it>are tedious and time consuming. In this study we have compared the conventional multi-step cloning technique with two knockout strategies that have been proven to work in other organisms, one-step-PCR- and Multisite Gateway-based systems.</p> <p>Results</p> <p>While the one-step-PCR strategy was found to be the fastest method for production of knockout constructs, it does not efficiently target genes of interest using gene-specific sequences of less than 80 nucleotides. Alternatively, the Multisite Gateway based approach is less time-consuming than conventional methods and is able to efficiently and reproducibly delete target genes.</p> <p>Conclusion</p> <p>Using the Multisite Gateway strategy, we have rapidly produced constructs that successfully produce specific gene deletions in epimastigotes of <it>T. cruzi</it>. This methodology should greatly facilitate reverse genetic studies in <it>T. cruzi</it>.</p
Chagas Disease and the London Declaration on Neglected Tropical Diseases
Fil: Tarleton, Rick L.. University of Georgia; Estados Unidos. Chagas Disease Foundation; Estados UnidosFil: Gurtler, Ricardo Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Urbina, Julio A.. Instituto Venezolano de Investigaciones Científicas; VenezuelaFil: Ramsey, Janine. Instituto Nacional de Salud Pública; MéxicoFil: Viotti, Rodolfo Jorge. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; Argentin
Spontaneous dormancy protects Trypanosoma cruzi during extended drug exposure
The ability of the Chagas disease agent Trypanosoma cruzi to resist extended in vivo exposure to highly effective trypanocidal compounds prompted us to explore the potential for dormancy and its contribution to failed drug treatments in this infection. We document the development of non-proliferating intracellular amastigotes in vivo and in vitro in the absence of drug treatment. Non-proliferative amastigotes ultimately converted to trypomastigotes and established infections in new host cells. Most significantly, dormant amastigotes were uniquely resistant to extended drug treatment in vivo and in vitro and could re-establish a flourishing infection after as many as 30 days of drug exposure. These results demonstrate a dormancy state in T. cruzi that accounts for the failure of highly cytotoxic compounds to completely resolve the infection. The ability of T. cruzi to establish dormancy throws into question current methods for identifying curative drugs but also suggests alternative therapeutic approaches.Fil: Sánchez Valdéz, Fernando Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina. University of Georgia; Estados UnidosFil: Poveda Padilla, Angélica Gabriela. University of Georgia; Estados UnidosFil: Wang, Wei. University of Georgia; Estados UnidosFil: Orr, Dylan. University of Georgia; Estados UnidosFil: Tarleton, Rick L.. University of Georgia; Estados Unido
Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
Reliable detection of Trypanosoma cruzi (T. cruzi) in vivo infections have long been needed to understand the complex biology of Chagas disease and to accurately evaluate the outcome of treatment regimens. Here, an integrated pipeline for automated quantification of T. cruzi-infected cells in 3D-reconstructed, cleared organs was developed. Light-sheet fluorescent microscopy allows us to accurately visualize and quantify not only actively proliferating but also dormant T. cruzi parasites and immune effector cells in whole-organs or tissues. Also, CUBIC-HistoVision pipeline to obtain uniform labeling of cleared organs with antibodies and nuclear stains was successfully adopted. Tissue clearing coupled to 3D immunostaining provides an unbiased approach to comprehensively evaluate drug treatment protocols, improve the understanding of the cellular organization of T. cruzi-infected tissues and is expected to advance discoveries related to anti-T. cruzi immune responses, tissue damage and repair in Chagas disease.Fil: Sánchez Valdéz, Fernando Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; ArgentinaFil: Padilla, Angel Marcelo. University of Georgia; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bustamante, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Georgia; Estados UnidosFil: Hawkins, Caleb W. D.. University of Georgia; Estados UnidosFil: Tarleton, Rick L.. University of Georgia; Estados Unido
A semi-quantitative GeLC-MS analysis of temporal proteome expression in the emerging nosocomial pathogen Ochrobactrum anthropi
A semi-quantitative gel-based analysis identifies distinct proteomic profiles associated with specific growth points for the nosocomial pathogen Ochrobactrum anthropi
Kinetoplastids:related protozoan pathogens, different diseases
Kinetoplastids are a group of flagellated protozoans that include the species Trypanosoma and Leishmania, which are human pathogens with devastating health and economic effects. The sequencing of the genomes of some of these species has highlighted their genetic relatedness and underlined differences in the diseases that they cause. As we discuss in this Review, steady progress using a combination of molecular, genetic, immunologic, and clinical approaches has substantially increased understanding of these pathogens and important aspects of the diseases that they cause. Consequently, the paths for developing additional measures to control these “neglected diseases” are becoming increasingly clear, and we believe that the opportunities for developing the drugs, diagnostics, vaccines, and other tools necessary to expand the armamentarium to combat these diseases have never been better
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