Caracterização de antígenos componentes de complexos ribonucleoprotéicos de Trypanosoma cruzi contendo seqüências com repetições de aminoácidos

Exportado OPUSMade available in DSpace on 2019-08-12T03:59:26Z (GMT). No. of bitstreams: 1 fabiano.pdf: 10118 bytes, checksum: dfdf2d8aee58adc11410e99a6d98c1f7 (MD5) Previous issue date: 17O Trypanosoma cruzi expressa diversos antígenos contendo seqüências com aminoácidos repetitivos. Neste trabalho foram caracterizados os antígenos TcRpL7a e TcRBP28 que contém repetições de aminoácidos semelhantes e apresentam similaridade com a proteína ribossômica L7a de eucariotos e com proteínas de ligação a RNA de T. brucei respectivamente. Ensaios de Western blot usando a proteína TcRpL7a completa e truncada indicaram que a resposta humoral dos pacientes é exclusiva contra a região repetitiva. Uma análise das seqüências de TcRpL7a de seis cepas diferentes do parasito indicaram que o número de módulos repetitivos é maior em cepas pertencentes à linhagem II de T. cruzi. 73% de um painel de 59 pacientes chagásicos apresentaram reatividade contra TcRpL7a, 71% contra TcRBP28 e 80% contra uma mistura dos dois antígenos. Um peptídeo sintético correspondente a uma parte da região repetitiva de TcRpL7a apresentou 46% de reatividade com os soros. Anticorpos gerados em camundongos identificaram as proteínas nativas presentes em níveis similares em todas as formas do ciclo de vida do parasito. Análises de frações subcelulares indicaram que TcRBP28 está presente no citoplasma e que TcRpL7a co-precipita com a fração de polissomos. A fusão com GFP confirmou a localização citoplasmática de TcRBP28 e mostrou um acúmulo da proteína GFP::TcRpL7a no núcleo da célula onde ocorre a biogênese do ribossomo. Camundongos imunizados com TcRpL7a::his desenvolveram anticorpos anti-IgG1/2a e apresentaram maior número de células secretoras de IFN-g. A capacidade de TcRpL7a de conferir proteção está sendo avaliada no presente momento.Trypanosoma cruzi expresses several proteins containing highly antigenic amino acid repeats. Here we characterized TcRpL7a and TcRBP28, which carry similar repeat motifs and share homology to the eukaryotic L7a ribosomal protein and to a T. brucei RNA binding protein, respectively. Analyses of the full length and truncated recombinant TcRpL7a showed that the humoral response of patients with Chagas disease is directed towards its repetitive domain. Sequence analyses of copies of TcRpL7a genes present in the genome of six T. cruzi strains indicate that the number of repeats is higher in T. cruzi II than in T. cruzi I strains. A serum panel of 59 patients showed that 73% reacted with TcRpL7a, 71% reacted with TcRBP28 and 80% reacted with 1:1 mixture of both antigens. Synthetic peptide harboring the TcRpL7a repeat motif reacted with 46% of chagasic sera. Antibodies raised against both antigens identified equivalent amounts of the native proteins in all three stages of the parasite life cycle. Analyses of subcellular fractions indicated that TcRBP28 is present in the cytoplasm whereas TcRpL7a co-fractionates with polysomes. Confirming their predicted cellular localization, GFP fusions showed that, whereas GFP::TcRBP28 localizes in the cytoplasm, GFP::TcRpL7a accumulates in the nucleus, where ribosome biogenesis occurs. Mice immunized with TcRpL7a::his presented high levels of IgG1 and IgG2a antibodies and INF-g secreting cells as well. Ongoing experiments are being done to evaluate the capacity of TcRpL7a to generate protection in mice against parasite infection

Assessing the efficiency of multiple sequence alignment programs.

Submitted by Nuzia Santos ([email protected]) on 2015-02-04T12:24:24Z No. of bitstreams: 1 2014_035.pdf: 516742 bytes, checksum: 2fa775121df1fd4b23c2255d7c7684fc (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-02-04T12:24:34Z (GMT) No. of bitstreams: 1 2014_035.pdf: 516742 bytes, checksum: 2fa775121df1fd4b23c2255d7c7684fc (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-02-04T12:30:02Z (GMT) No. of bitstreams: 1 2014_035.pdf: 516742 bytes, checksum: 2fa775121df1fd4b23c2255d7c7684fc (MD5)Made available in DSpace on 2015-02-04T12:30:02Z (GMT). No. of bitstreams: 1 2014_035.pdf: 516742 bytes, checksum: 2fa775121df1fd4b23c2255d7c7684fc (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Centro de Excelência em Bioinformática. Belo Horizonte, MG, Brazil/Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou Grupo de Genômica Biologia Computacional. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Centro de Excelência em Bioinformática. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Centro de Excelência em Bioinformática. Belo Horizonte, MG, Brazil/Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou Grupo de Genômica Biologia Computacional. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Centro de Excelência em Bioinformática. Belo Horizonte, MG, Brazil/Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou Grupo de Genômica Biologia Computacional. Belo Horizonte, MG, BrazilBACKGROUND: Multiple sequence alignment (MSA) is an extremely useful tool for molecular and evolutionary biology and there are several programs and algorithms available for this purpose. Although previous studies have compared the alignment accuracy of different MSA programs, their computational time and memory usage have not been systematically evaluated. Given the unprecedented amount of data produced by next generation deep sequencing platforms, and increasing demand for large-scale data analysis, it is imperative to optimize the application of software. Therefore, a balance between alignment accuracy and computational cost has become a critical indicator of the most suitable MSA program. We compared both accuracy and cost of nine popular MSA programs, namely CLUSTALW, CLUSTAL OMEGA, DIALIGN-TX, MAFFT, MUSCLE, POA, Probalign, Probcons and T-Coffee, against the benchmark alignment dataset BAliBASE and discuss the relevance of some implementations embedded in each program's algorithm. Accuracy of alignment was calculated with the two standard scoring functions provided by BAliBASE, the sum-of-pairs and total-column scores, and computational costs were determined by collecting peak memory usage and time of execution. RESULTS: Our results indicate that mostly the consistency-based programs Probcons, T-Coffee, Probalign and MAFFT outperformed the other programs in accuracy. Whenever sequences with large N/C terminal extensions were present in the BAliBASE suite, Probalign, MAFFT and also CLUSTAL OMEGA outperformed Probcons and T-Coffee. The drawback of these programs is that they are more memory-greedy and slower than POA, CLUSTALW, DIALIGN-TX, and MUSCLE. CLUSTALW and MUSCLE were the fastest programs, being CLUSTALW the least RAM memory demanding program. CONCLUSIONS: Based on the results presented herein, all four programs Probcons, T-Coffee, Probalign and MAFFT are well recommended for better accuracy of multiple sequence alignments. T-Coffee and recent versions of MAFFT can deliver faster and reliable alignments, which are specially suited for larger datasets than those encountered in the BAliBASE suite, if multi-core computers are available. In fact, parallelization of alignme

Revisitingmolecular serotyping of Streptococcus pneumonia

Submitted by Nuzia Santos ([email protected]) on 2016-07-06T11:40:35Z No. of bitstreams: 1 ve_Dhian_Camargo_ Revisitingmolecular_CPqRR_2015.pdf: 1647355 bytes, checksum: d446af4ae8ae1cdd1b0e0853582ff5ec (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2016-07-06T11:43:26Z (GMT) No. of bitstreams: 1 ve_Dhian_Camargo_ Revisitingmolecular_CPqRR_2015.pdf: 1647355 bytes, checksum: d446af4ae8ae1cdd1b0e0853582ff5ec (MD5)Made available in DSpace on 2016-07-06T11:43:26Z (GMT). No. of bitstreams: 1 ve_Dhian_Camargo_ Revisitingmolecular_CPqRR_2015.pdf: 1647355 bytes, checksum: d446af4ae8ae1cdd1b0e0853582ff5ec (MD5) Previous issue date: 2015Made available in DSpace on 2016-07-14T19:10:31Z (GMT). No. of bitstreams: 3 ve_Dhian_Camargo_ Revisitingmolecular_CPqRR_2015.pdf.txt: 41150 bytes, checksum: 677ee69411513ac8a87f484354a4dac8 (MD5) ve_Dhian_Camargo_ Revisitingmolecular_CPqRR_2015.pdf: 1647355 bytes, checksum: d446af4ae8ae1cdd1b0e0853582ff5ec (MD5) license.txt: 2991 bytes, checksum: 5a560609d32a3863062d77ff32785d58 (MD5) Previous issue date: 2015Fundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, Brasil / Fundação Ezequiel Dias. Serviço de Bactérias e Doenças de Fungos. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Grupo de Biologia e Genômica Computacional. Belo Horizonte, MG, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Centro de Excelência em Bioinformática. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Grupo de Biologia e Genômica Computacional. Belo Horizonte, MG, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Centro de Excelência em Bioinformática. Belo Horizonte, MG, Brasil.Fundação Ezequiel Dias. Serviço de Bactérias e Doenças de Fungos. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, Brasil.Background: Ninety-two Streptococcus pneumoniae serotypes have been described so far, but the pneumococcal conjugate vaccine introduced in the Brazilian basic vaccination schedule in 2010 covers only the ten most prevalent in the country. Pneumococcal serotype-shifting after massive immunization is a major concern and monitoring this phenomenon requires efficient and accessible serotyping methods. Pneumococcal serotyping based on antisera produced in animals is laborious and restricted to a few reference laboratories. Alternatively, molecular serotyping methods assess polymorphisms in the cps gene cluster, which encodes key enzymes for capsular polysaccharides synthesis in pneumococci. In one such approach, cps-RFLP, the PCR amplified cps loci are digested with an endonuclease, generating serotype-specific fingerprints on agarose gel electrophoresis. Methods: In this work, in silico and in vitro approaches were combined to demonstrate that XhoII is the most discriminating endonuclease for cps-RFLP, and to build a database of serotype-specific fingerprints that accommodates the genetic diversity within the cps locus of 92 known pneumococci serotypes. Results: The expected specificity of cps-RFLP using XhoII was 76% for serotyping and 100% for serogrouping. The database of cps-RFLP fingerprints was integrated to Molecular Serotyping Tool (MST), a previously published web-based software for molecular serotyping. In addition, 43 isolates representing 29 serotypes prevalent in the state of Minas Gerais, Brazil, from 2007 to 2013, were examined in vitro; 11 serotypes (nine serogroups) matched the respective in silico patterns calculated for reference strains. The remaining experimental patterns, despite their resemblance to their expected in silico patterns, did not reach the threshold of similarity score to be considered a match and were then added to the database. Conclusion: The cps-RFLP method with XhoII outperformed the antisera-based and other molecular serotyping methods in regard of the expected specificity. In order to accommodate the genetic variability of the pneumococci cps loci, the database of cps-RFLP patterns will be progressively expanded to include new variant in vitro patterns. The cps-RFLP method with endonuclease XhoII coupled with MST for computer-assisted interpretation of results may represent a relevant contribution to the real time detection of changes in regional pneumococci population diversity in response to mass immunization programs

Evolutionary analysis of the cystatin family in three Schistosoma species.

Submitted by Nuzia Santos ([email protected]) on 2015-07-10T12:30:46Z No. of bitstreams: 1 2014_129.pdf: 2673173 bytes, checksum: f82bf3993fce5f7832126b14568b22be (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-07-10T12:30:53Z (GMT) No. of bitstreams: 1 2014_129.pdf: 2673173 bytes, checksum: f82bf3993fce5f7832126b14568b22be (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-07-10T12:36:43Z (GMT) No. of bitstreams: 1 2014_129.pdf: 2673173 bytes, checksum: f82bf3993fce5f7832126b14568b22be (MD5)Made available in DSpace on 2015-07-10T12:36:43Z (GMT). No. of bitstreams: 1 2014_129.pdf: 2673173 bytes, checksum: f82bf3993fce5f7832126b14568b22be (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Genômica e Biologia Computacional. Centro de Excelência em Bioinformática. Belo Horizonte, MG, Brasil /Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais. Belo Horizonte, MG, Brasil/ Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Genômica e Biologia Computacional. Centro de Excelência em Bioinformática. Belo Horizonte, MG, Brasil /Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais. Belo Horizonte, MG, Brasil/ Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Genômica e Biologia Computacional. Centro de Excelência em Bioinformática. Belo Horizonte, MG, Brasil /Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais. Belo Horizonte, MG, Brasil/Faculdade Infórium de Tecnologia, Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Genômica e Biologia Computacional. Centro de Excelência em Bioinformática. Belo Horizonte, MG, Brasil /Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Genômica e Biologia Computacional. Centro de Excelência em Bioinformática. Belo Horizonte, MG, Brasil /Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais. Belo Horizonte, MG, Brasil/Faculdade Infórium de Tecnologia, Belo Horizonte, MG, BrazilThe cystatin family comprises cysteine protease inhibitors distributed in 3 subfamilies (I25A-C). Family members lacking cystatin activity are currently unclassified. Little is known about the evolution of Schistosoma cystatins, their physiological roles, and expression patterns in the parasite life cycle. The present study aimed to identify cystatin homologs in the predicted proteome of three Schistosoma species and other Platyhelminthes. We analyzed the amino acid sequence diversity focused in the identification of protein signatures and to establish evolutionary relationships among Schistosoma and experimentally validated human cystatins. Gene expression patterns were obtained from different developmental stages in Schistosoma mansoni using microarray data. In Schistosoma, only I25A and I25B proteins were identified, reflecting little functional diversification. I25C and unclassified subfamily members were not identified in platyhelminth species here analyzed. The resulting phylogeny placed cystatins in different clades, reflecting their molecular diversity. Our findings suggest that Schistosoma cystatins are very divergent from their human homologs, especially regarding the I25B subfamily. Schistosoma cystatins also differ significantly from other platyhelminth homologs. Finally, transcriptome data publicly available indicated that I25A and I25B genes are constitutively expressed thus could be essential for schistosome life cycle progression. In summary, this study provides insights into the evolution, classification, and functional diversification of cystatins in Schistosoma and other Platyhelminthes, improving our understanding of parasite biology and opening new frontiers in the identification of novel therapeutic targets against helminthiases

Global characterization of gene expression in the brain of starved immature Rhodnius prolixus

Background Rhodnius prolixus is a vector of Chagas disease and has become a model organism to study physiology, behavior, and pathogen interaction. The publication of its genome allowed initiating a process of comparative characterization of the gene expression profiles of diverse organs exposed to varying conditions. Brain processes control the expression of behavior and, as such, mediate immediate adjustment to a changing environment, allowing organisms to maximize their chances to survive and reproduce. The expression of fundamental behavioral processes like feeding requires fine control in triatomines because they obtain their blood meals from potential predators. Therefore, the characterization of gene expression profiles of key components modulating behavior in brain processes, like those of neuropeptide precursors and their receptors, seems fundamental. Here we study global gene expression profiles in the brain of starved R. prolixus fifth instar nymphs by means of RNA sequencing (RNA-Seq). Results The expression of neuromodulatory genes such as those of precursors of neuropeptides, neurohormones, and their receptors; as well as the enzymes involved in the biosynthesis and processing of neuropeptides and biogenic amines were fully characterized. Other important gene targets such as neurotransmitter receptors, nuclear receptors, clock genes, sensory receptors, and takeouts genes were identified and their gene expression analyzed. Conclusion We propose that the set of neuromodulatory-related genes highly expressed in the brain of starved R. prolixus nymphs deserves functional characterization to allow the subsequent development of tools targeting them for bug control. As the brain is a complex structure that presents functionally specialized areas, future studies should focus on characterizing gene expression profiles in target areas, e.g. mushroom bodies, to complement our current knowledge.Fil: Coraiola Nevoa, Jessica. Fundación Oswaldo Cruz; BrasilFil: Latorre Estivalis, Jose Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Sviatopolk Mirsky Pais, Fabiano. Fundación Oswaldo Cruz; BrasilFil: Pinto Marliere, Newmar. Fundación Oswaldo Cruz; BrasilFil: da Rocha Fernandes, Gabriel. Fundación Oswaldo Cruz; BrasilFil: Lorenzo, Marcelo Gustavo. Fundación Oswaldo Cruz; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Aparecida Guarneri, Alessandra. Fundación Oswaldo Cruz; Brasi

Overexpression of eukaryotic initiation factor 5A (eIF5A) affects susceptibility to benznidazole in Trypanosoma cruzi populations

Eukaryotic initiation factor 5A (eIF5A) is a conserved protein with an essential role in translation elongation. Using one and two-dimensional western blotting, we showed that the eIF5A protein level was 2-fold lower in benznidazole (BZ)-resistant (BZR and 17LER) Trypanosoma cruzi populations than in their respective susceptible counterparts (BZS and 17WTS). To confirm the role of eIF5A in BZ resistance, we transfected BZS and 17WTS with the wild-type eIF5A or mutant eIF5A-S2A (in which serine 2 was replaced by alanine). Upon overexpressing eIF5A, both susceptible lines became approximately 3- and 5-fold more sensitive to BZ. In contrast, the eIF5A-S2A mutant did not alter its susceptibility to BZ. These data suggest that BZ resistance might arise from either decreasing the translation of proteins that require eIF5A, or as a consequence of differential levels of precursors for the hypusination reactions (e.g., spermidine and trypanothione), both of which alter BZ’s effects in the parasite

ZIKV – CDB: A Collaborative Database to Guide Research Linking SncRNAs and ZIKA Virus Disease Symptoms

Background: In early 2015, a ZIKA Virus (ZIKV) infection outbreak was recognized in northeast Brazil, where concerns over its possible links with infant microcephaly have been discussed. Providing a causal link between ZIKV infection and birth defects is still a challenge. MicroRNAs (miRNAs) are small noncoding RNAs (sncRNAs) that regulate post-transcriptional gene expression by translational repression, and play important roles in viral pathogenesis and brain development. The potential for flavivirus-mediated miRNA signalling dysfunction in brain-tissue development provides a compelling hypothesis to test the perceived link between ZIKV and microcephaly. Methodology/Principal Findings: Here, we applied in silico analyses to provide novel insights to understand how Congenital ZIKA Syndrome symptoms may be related to an imbalance in miRNAs function. Moreover, following World Health Organization (WHO) recommendations, we have assembled a database to help target investigations of the possible relationship between ZIKV symptoms and miRNA-mediated human gene expression. Conclusions/Significance: We have computationally predicted both miRNAs encoded by ZIKV able to target genes in the human genome and cellular (human) miRNAs capable of interacting with ZIKV genomes. Our results represent a step forward in the ZIKV studies, providing new insights to support research in this field and identify potential targets for therapy.</p

ZIKV – CDB: A Collaborative Database to Guide Research Linking SncRNAs and ZIKA Virus Disease Symptoms

Submitted by Nuzia Santos ([email protected]) on 2016-08-02T19:23:10Z No. of bitstreams: 1 Pylro_Victor_ZIKV – CDB_CPqRR_2016.pdf: 339073 bytes, checksum: ecc68d1dda0f81065b378c9d58ecc051 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2016-08-02T19:30:02Z (GMT) No. of bitstreams: 1 Pylro_Victor_ZIKV – CDB_CPqRR_2016.pdf: 339073 bytes, checksum: ecc68d1dda0f81065b378c9d58ecc051 (MD5)Made available in DSpace on 2016-08-02T19:30:02Z (GMT). No. of bitstreams: 1 Pylro_Victor_ZIKV – CDB_CPqRR_2016.pdf: 339073 bytes, checksum: ecc68d1dda0f81065b378c9d58ecc051 (MD5) Previous issue date: 2016FAPEMIG, CNPq, CAPES, FSMPFundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Informática de Biossistemas e Grupo de Genomica. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Informática de Biossistemas e Grupo de Genomica. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Informática de Biossistemas e Grupo de Genomica. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Informática de Biossistemas e Grupo de Genomica. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Informática de Biossistemas e Grupo de Genomica. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Informática de Biossistemas e Grupo de Genomica. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Informática de Biossistemas e Grupo de Genomica. Belo Horizonte, MG, Brasil.University of Chicago. Department of Ecology and Evolution.Chicago, IL, United States of America/Institute for Genomic and Systems Biology. Argonne National Laboratory. Argonne, IL, United States of America/Department of Surgery, The University of Chicago, Chicago, Illinois, United States of America.Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Informática de Biossistemas e Grupo de Genomica. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Informática de Biossistemas e Grupo de Genomica. Belo Horizonte, MG, Brasil.BACKGROUND: In early 2015, a ZIKA Virus (ZIKV) infection outbreak was recognized in northeast Brazil, where concerns over its possible links with infant microcephaly have been discussed. Providing a causal link between ZIKV infection and birth defects is still a challenge. MicroRNAs (miRNAs) are small noncoding RNAs (sncRNAs) that regulate post-transcriptional gene expression by translational repression, and play important roles in viral pathogenesis and brain development. The potential for flavivirus-mediated miRNA signalling dysfunction in brain-tissue development provides a compelling hypothesis to test the perceived link between ZIKV and microcephaly. METHODOLOGY/PRINCIPAL FINDINGS: Here, we applied in silico analyses to provide novel insights to understand how Congenital ZIKA Syndrome symptoms may be related to an imbalance in miRNAs function. Moreover, following World Health Organization (WHO) recommendations, we have assembled a database to help target investigations of the possible relationship between ZIKV symptoms and miRNA-mediated human gene expression. CONCLUSIONS/SIGNIFICANCE: We have computationally predicted both miRNAs encoded by ZIKV able to target genes in the human genome and cellular (human) miRNAs capable of interacting with ZIKV genomes. Our results represent a step forward in the ZIKV studies, providing new insights to support research in this field and identify potential targets for therap

Genetic homogeneity among Leishmania (Leishmania) infantum isolates from dog and human samples in Belo Horizonte Metropolitan Area (BHMA); Minas Gerais; Brazil

Submitted by Nuzia Santos ([email protected]) on 2016-01-29T17:47:35Z No. of bitstreams: 1 Genetic homogeneity among Leishmania (Leishmania) infantum isolates.pdf: 10721186 bytes, checksum: 6344edd6cdea306724b13c911ba8a110 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2016-02-01T11:46:00Z (GMT) No. of bitstreams: 1 Genetic homogeneity among Leishmania (Leishmania) infantum isolates.pdf: 10721186 bytes, checksum: 6344edd6cdea306724b13c911ba8a110 (MD5)Made available in DSpace on 2016-02-01T11:46:00Z (GMT). No. of bitstreams: 1 Genetic homogeneity among Leishmania (Leishmania) infantum isolates.pdf: 10721186 bytes, checksum: 6344edd6cdea306724b13c911ba8a110 (MD5) Previous issue date: 2015Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Pesquisas Clínicas. Belo Horizonte, MG, Brasil/Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Laboratório de Epidemiologia das Doenças Infecciosas e Parasitárias. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Pesquisas Clínicas. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Pesquisas Clínicas. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Laboratório de Epidemiologia das Doenças Infecciosas e Parasitárias. Belo Horizonte, MG, Brasil/Universidade Federal de Minas Gerais. Faculdade de Medicina. Pós-graduação em Ciências da Saúde: Infectologia e Medicina Tropical. Belo Horizonte, MG, Brasil/Universidade Federal de Ouro Preto. Escola de Farmácia. Laboratório de Pesquisas Clínicas. Ouro Preto, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Laboratório de Toxoplasmose Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Genômica e Biologia Computacional. Belo Horizonte, MG, Brasil.Universidade Federal de Ouro Preto. Escola de Farmácia. Laboratório de Pesquisas Clínicas. Ouro Preto, MG, Brasil.Universidade Federal de Ouro Preto. Escola de Farmácia. Laboratório de Pesquisas Clínicas. Ouro Preto, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Pesquisas Clínicas. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Laboratório de Epidemiologia das Doenças Infecciosas e Parasitárias. Belo Horizonte, MG, Brasil/Universidade Federal de Minas Gerais. Faculdade de Medicina. Pós-graduação em Ciências da Saúde: Infectologia e Medicina Tropical. Belo Horizonte, MG, Brasil.BACKGROUND: Certain municipalities in the Belo Horizonte Metropolitan Area (BHMA), Minas Gerais, Brazil, have the highest human visceral leishmaniasis (VL) mortality rates in the country and also demonstrate high canine seropositivity. In Brazil, the etiologic agent of VL is Leishmania (Leishmania) infantum. The aim of this study was to evaluate the intraspecific genetic variability of parasites from humans and from dogs with different clinical forms of VL in five municipalities of BHMA using PCR-RFLP and two target genes: kinetoplast DNA (kDNA) and gp63. METHODS: In total, 45 samples of DNA extracted from clinical samples (n = 35) or L. infantum culture (n = 10) were evaluated. These samples originated from three groups: adults (with or without Leishmania/HIV co-infection; n = 14), children (n = 18) and dogs (n = 13). The samples were amplified for the kDNA target using the MC1 and MC2 primers (447 bp), while the Sg1 and Sg2 (1330 bp) primers were used for the gp63 glycoprotein target gene. RESULTS: The restriction enzyme patterns of all the samples tested were monomorphic. CONCLUSIONS: These findings reveal a high degree of genetic homogeneity for the evaluated gene targets among L. infantum samples isolated from different hosts and representing different clinical forms of VL in the municipalities of BHMA studied

Genetic homogeneity among Leishmania (Leishmania) infantum isolates from dog and human samples in Belo Horizonte Metropolitan Area (BHMA), Minas Gerais, Brazil.

Background: Certain municipalities in the Belo Horizonte Metropolitan Area (BHMA), Minas Gerais, Brazil, have the highest human visceral leishmaniasis (VL) mortality rates in the country and also demonstrate high canine seropositivity. In Brazil, the etiologic agent of VL is Leishmania (Leishmania) infantum. The aim of this study was to evaluate the intraspecific genetic variability of parasites from humans and from dogs with different clinical forms of VL in five municipalities of BHMA using PCR-RFLP and two target genes: kinetoplast DNA (kDNA) and gp63. Methods: In total, 45 samples of DNA extracted from clinical samples (n = 35) or L. infantum culture (n = 10) were evaluated. These samples originated from three groups: adults (with or without Leishmania/HIV co-infection; n = 14), children (n = 18) and dogs (n = 13). The samples were amplified for the kDNA target using the MC1 and MC2 primers (447 bp), while the Sg1 and Sg2 (1330 bp) primers were used for the gp63 glycoprotein target gene. Results: The restriction enzyme patterns of all the samples tested were monomorphic. Conclusions: These findings reveal a high degree of genetic homogeneity for the evaluated gene targets among L. infantum samples isolated from different hosts and representing different clinical forms of VL in the municipalities of BHMA studied