CA88, a nuclear repetitive DNA sequence identified in Schistosoma mansoni, aids in the genotyping of nine Schistosoma species of medical and veterinary importance

CA88 is the first long nuclear repetitive DNA sequence identified in the blood fluke, Schistosoma mansoni. The assembled S. mansoni sequence, which contains the CA88 repeat, has 8,887 nucleotides and at least three repeat units of approximately 360 bp. In addition, CA88 also possesses an internal CA microsatellite, identified as SmBr18. Both PCR and BLAST analysis have been used to analyse and confirm the CA88 sequence in other S. mansoni sequences in the public database. PCR-acquired nuclear repetitive DNA sequence profiles from nine Schistosoma species were used to classify this organism into four genotypes. Included among the nine species analysed were five sequences of both African and Asian lineages that are known to infect humans. Within these genotypes, three of them refer to recognised species groups. A panel of four microsatellite loci, including SmBr18 and three previously published loci, has been used to characterise the nine Schistosoma species. Each species has been identified and classified based on its CA88 DNA fingerprint profile. Furthermore, microsatellite sequences and intra-specific variation have also been observed within the nine Schistosoma species sequences. Taken together, these results support the use of these markers in studying the population dynamics of Schistosoma isolates from endemic areas and also provide new methods for investigating the relationships between different populations of parasites. In addition, these data also indicate that Schistosoma magrebowiei is not a sister taxon to Schistosoma mattheei, prompting a new designation to a basal clade.Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Imunologia e ParasitologiaInstituto de Pesquisas René Rachou-FiocruzThe Natural History Museum Department of ZoologySanta Casa de Belo Horizonte Programa de Pós-Graduação e PesquisaUniversidade Federal de Ouro Preto Escola de Farmácia Laboratório de Pesquisas ClínicasUNIFESP, EPM, Imunologia e ParasitologiaSciEL

Unlocking the in vitro anti-Trypanosoma cruzi activity of halophyte plants from the southern Portugal

Objective: To evaluate the in vitro anti-Trypanosoma cruzi (T. cruzi) activity of organic extracts prepared from halophyte species collected in the southern coast of Portugal (Algarve), and chemically characterize the most active samples.Methods: Acetone, dichloromethane and methanol extracts were prepared from 31 halophyte species and tested in vitro against trypomastigotes and intracellular amastigotes of the Tulahuen strain of T. cruzi. The most active extract was fractionated by preparative HPLC-DAD, affording 11 fractions. The most selective fraction was fully characterized by H-1 NMR.Results: From 94 samples tested, one was active, namely the root dichloromethane extract of Juncus acutus (IC50 < 20 mu g/mL). This extract was fractionated by HPLC, affording 11 fractions, one of them containing only a pure compound (juncunol), and tested for anti-parasitic activity. Fraction 8 (IC50 = 4.1 mu g/mL) was the most active, and was further characterized by H-1 NMR. The major compounds were phenanthrenes, 9,10-dihydrophenanthrenes and benzocoumarins.Conclusion: Our results suggest that the compounds identified in fraction 8 are likely responsible for the observed anti parasitic activity. Further research is in progress aiming to isolate and identify the specific active molecules. To the best of our knowledge, this is the first report on the in vitro anti T. cruzi activity of halophyte species.PROEP/CNPq/FIOCRUZ 401988/2012-0; FCT/CAPES 2358, 2014/201

Genome of the Avirulent Human-Infective Trypanosome—Trypanosoma rangeli

Background: Trypanosoma rangeli is a hemoflagellate protozoan parasite infecting humans and other wild and domestic mammals across Central and South America. It does not cause human disease, but it can be mistaken for the etiologic agent of Chagas disease, Trypanosoma cruzi. We have sequenced the T. rangeli genome to provide new tools for elucidating the distinct and intriguing biology of this species and the key pathways related to interaction with its arthropod and mammalian hosts.  Methodology/Principal Findings: The T. rangeli haploid genome is ,24 Mb in length, and is the smallest and least repetitive trypanosomatid genome sequenced thus far. This parasite genome has shorter subtelomeric sequences compared to those of T. cruzi and T. brucei; displays intraspecific karyotype variability and lacks minichromosomes. Of the predicted 7,613 protein coding sequences, functional annotations could be determined for 2,415, while 5,043 are hypothetical proteins, some with evidence of protein expression. 7,101 genes (93%) are shared with other trypanosomatids that infect humans. An ortholog of the dcl2 gene involved in the T. brucei RNAi pathway was found in T. rangeli, but the RNAi machinery is non-functional since the other genes in this pathway are pseudogenized. T. rangeli is highly susceptible to oxidative stress, a phenotype that may be explained by a smaller number of anti-oxidant defense enzymes and heatshock proteins.  Conclusions/Significance: Phylogenetic comparison of nuclear and mitochondrial genes indicates that T. rangeli and T. cruzi are equidistant from T. brucei. In addition to revealing new aspects of trypanosome co-evolution within the vertebrate and invertebrate hosts, comparative genomic analysis with pathogenic trypanosomatids provides valuable new information that can be further explored with the aim of developing better diagnostic tools and/or therapeutic targets

Inter-relações entre os ciclos de transmissão do Trypanosoma cruzi no município de Bambuí, Minas Gerais, Brasil Relationships between Trypanosoma cruzi transmission cycles in the county of Bambuí, Minas Gerais, Brazil

Neste trabalho apresentamos um estudo das inter-relações recentes entre os ciclos de transmissão silvestre e doméstico do T. cruzi no município de Bambuí, Minas Gerais, Brasil. No final da década de 1930, o Panstrongylus megistus era encontrado em 75% das casas. Subseqüentemente, o Triatoma infestans tornou-se a espécie predominante, sendo encontrada em 20% das residências urbanas e em mais de 60% das periurbanas. Com as intensas campanhas de borrifação desenvolvidas entre 1956 e 1969, o T. infestans foi erradicado do município, e a transmissão da doença de Chagas ao homem, interrompida, com aparecimento de P. megistus em residências rurais. Amostras de T. cruzi isoladas via xenodiagnóstico e hemocultura de 43 gambás (Didelphis albiventris) capturados em ambiente peridomiciliar e silvestre foram caracterizadas isoenzimaticamente e, independentemente da via de isolamento, apresentaram perfil de zimodema Z1. Por meio do Programa de Vigilância Epidemiológica da doença de Chagas, no período de agosto de 1986 a dezembro de 1988, 154 exemplares de P. megistus foram capturados pela população no peridomicílio e intradomicílio rural, estando 9,8% infectados pelo T. cruzi. Na caracterização isoenzimática de 13 amostras de T. cruzi isoladas desses triatomíneos, seis pertenciam ao zimodema Z1 (ciclo de transmissão silvestre), e sete ao Z2 (ciclo de transmissão doméstico). A captura de exemplares de P. megistus no intradomicílio, naturalmente infectados com parasitas de ambos os ciclos, indica a superposição dos ciclos de transmissão da doença de Chagas no município de Bambuí. Outra evidência da inter-relação dos ciclos pôde ser observada no isolamento de T. cruzi Z2 de um gato e a participação do cão como reservatório de T. cruzi Z1. A presença do P. megistus no peridomicílio representa importante elo entre o ambiente silvestre e o intradomicílio, servindo como veiculador do T. cruzi Z1 e na manutenção de ciclos de transmissão do T. cruzi Z2 no peridomicílio e domicílio, propiciando de forma gradual a reinfestação do município caso a Vigilância Epidemiológica seja interrompida.This study examines recent relationships between domestic and sylvatic transmission cycles of T.cruzi in the county of Bambuí, MG, Brazil. In the late l930s, Panstrongylus megistus was found in 75% of houses. Subsequently, Triatoma infestans became the predominant species, found in 20% of urban households and in more than 60% of periurban homes. With intense insecticide control campaigns between l956 and l969, T. infestans was eradicated from the county, transmission of Chagas' disease to man was interrupted, and P. mesgistus appeared in rural residences. Samples of T. cruzi isolated by xenodiagnosis and hemoculture from 43 opossums (Didelphis albiventris) captured in both peridomiciliary and sylvatic areas were characterized by isoenzyme analysis and - regardless of isolation method - were found to present the Z1 zymodeme profile. Through the "Chagas' Disease Epidemiological Surveillance Program", from August l986 to December l988, 154 specimens of P. megistus were captured by the local population in both peridomiciliary and intradomiciliary environments, of which 9.8% were infected with T. cruzi. In isoenzyme analyses of l3 T. cruzi strains isolated from these triatomines, six were found to be of the Z1 zymodeme (sylvatic transmission cycles) and seven were found to be of the Z2 zymodeme (domestic transmission cycle). The capture of P. megistus specimens in intradomiciliary environments that were naturally infected with parasites of both cycle types indicates an overlap of transmission cycles of Chagas' disease in the county of Bambuí. Further evidence for the interrelationship of the two cycles was provided by the isolation of T. cruzi of the Z2 zymodeme from a cat and the participation of the dog as a reservoir of Z1 T. cruzi. The presence of P. megistus in the peridomiciliary environment represents an important link between the sylvatic and intradomestic environments serving as a carrier of Z1 T. cruzi and maintaining the transmission cycles of Z2 T. cruzi in the peridomestic and intradomestic environments thus providing the potential for a gradual reinfestation of the county if the "Epidemiological Surveillance" is interrupted

Evaluation of compliance through specific interviews: a prospective study of 73 children with acute lymphoblastic leukemia

Submitted by Nuzia Santos ([email protected]) on 2012-12-17T12:37:51Z No. of bitstreams: 1 58. Evaluation of compliance.pdf: 165285 bytes, checksum: 3cf90316b3dc77bd5a351fdf5237ee13 (MD5)Made available in DSpace on 2012-12-17T12:37:51Z (GMT). No. of bitstreams: 1 58. Evaluation of compliance.pdf: 165285 bytes, checksum: 3cf90316b3dc77bd5a351fdf5237ee13 (MD5) Previous issue date: 2005FAPEMIG, CNPq, FIOCRUZ, Pronex.Universidade Federal de Minas Gerais. School of Medicine. Department of Pediatrics. Belo Horizonte, MG, Brasil.Department of Pediatrics, School of Medicine, UFMG, Belo Horizonte, MG, Brazil.CNPq research scholarship holder.FAPEMIG research scholarship holder.Oswaldo Cruz Foundation. Research Center René Rachou. Belo Horizonte, MG, Brasil.Objective: To evaluate compliance in children with acute lymphoblastic leukemia. Method: Compliance was assessed through specific interviews. Results: A total of 73 patients, aged under 18 and who had concluded the maintenance phase of chemotherapy, were enrolled on the study. Eighty-one per cent of the interviews were conducted with the patients. mothers; 92% of the families stated that medical instructions had been understood well. Interviews indicated that 27% of the patients did not receive their medication twice or more during the maintenance phase, without medical direction for this. These children were considered non-compliant. Sixteen per cent of the children failed to receive their medication three times or more. The main reason for non-compliance was forgetfulness. In ten cases the reported dosage of drugs was not that which was prescribed. No significant associations of non-compliance with parents. schooling level, number of family members or per capita family income were detected. The 8.5-year estimated probability of event free survival was 72.4% (95% CI: 59.2-82.3). The event free survival curves for non-compliant children were not statistically different from those for the compliant group. Conclusions: Results suggest that comprehensive approaches to the problem of non-ompliance are urgently needed

Ocorrência de leishmaniose visceral canina autóctone em uma área não-endêmica: Bom Sucesso, Minas Gerais, Brasil

Submitted by Nuzia Santos ([email protected]) on 2013-07-12T18:35:19Z No. of bitstreams: 1 144. SILVA MR, MARQUES MJ.pdf: 169484 bytes, checksum: b16b0f1dc309e502eff490f0e3aeb9d8 (MD5)Made available in DSpace on 2013-07-12T18:35:19Z (GMT). No. of bitstreams: 1 144. SILVA MR, MARQUES MJ.pdf: 169484 bytes, checksum: b16b0f1dc309e502eff490f0e3aeb9d8 (MD5) Previous issue date: 2008Embrapa Gado de Leite. Núcleo de Qualidade do Leite, Saúde Animal e Socioeconomia. Juiz de Fora, Brasil / Universidade Federal de Minas Gerais. Faculdade de Medicina. Belo Horizonte, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Belo Horizonte, MG, Brasil.Universidade Federal de Lavras. Departamento de Medicina Veterinária. Lavras, Brasil.Universidade Federal de Ouro Preto. Instituto de Ciências Exatas e Biológicas. Ouro Preto, Brasil.Universidade Federal de Ouro Preto. Instituto de Ciências Exatas e Biológicas. Ouro Preto, Brasil.O presente trabalho descreve inicialmente um cão com sintomas característicos de leishmaniose visceral. Amostra de soro desse cão foi positiva por imunofluorescência indireta (IFI) conduzida no IgG total anti-Leishmania em 1999. Além disso, tecidos desse cão foram positivos por reação em cadeia pela polimerase (PCR) conduzida em 2004, identificando DNA de Leishmania no cerebelo, fígado, rim e intestino. Esta é a primeira vez que um cão com leishmaniose visceral autóctone foi descrito no Município de Bom Sucesso, Minas Gerais, Brasil. O achado desse cão reagente à IFI levou a uma investigação epidemiológica nesse município. Essa investigação foi conduzida de março de 1999 a dezembro de 2005. Vinte e dois de um total de 734 (3%) cães examinados foram reagentes à IFI. Entre os 22 cães IFI reagentes, seis apresentaram sintomas característicos de leishmaniose visceral canina. Os resultados desta investigação epidemiológica foram enviados às autoridades locais e estaduais de saúde pública requerendo medidas preventivas e de controle para leishmaniose visceral de forma a interromper a transmissão da doença e evitar a ocorrência de casos humanos.The article begins by describing a dog with characteristic symptoms of visceral leishmaniasis. A serum sample from this animal was positive by indirect immunofluorescence (IIF) performed in anti-Leishmania total IgG in 1999. Tissues from the same dog were also positive by polymerase chain reaction (PCR) in 2004, identifying Leishmania DNA in the cerebellum, liver, kidney, and intestine. This is the first report of a dog with autochthonous visceral leishmaniasis in the county of Bom Sucesso, Minas Gerais State, Brazil. The finding of this IIF-positive dog led to a canine visceral leishmaniasis epidemiological investigation in the county. The investigation was conducted from March 1999 to December 2005. IIF was positive for Leishmania in 22 (3%) of 734 examined dogs. Among the 22 IIF-positive dogs, six presented characteristic symptoms of canine visceral leishmaniasis. The results of this epidemiological investigation were sent to local and State public health authorities, requesting visceral leishmaniasis control and preventive measures to interrupt transmission of the disease and avoid the occurrence of human cases

First report on the occurrence of Trypanosoma rangeli Tejera, 1920 in the state of Ceará, Brazil, in naturally infected triatomine Rhodnius nasutus Stål, 1859 (Hemiptera, Reduviidae, Triatominae)

Submitted by Nuzia Santos ([email protected]) on 2013-06-21T12:59:28Z No. of bitstreams: 1 39 DIAS FBS.pdf: 125300 bytes, checksum: 372104899d268ec0b3afbe04e9d6f2fa (MD5)Made available in DSpace on 2013-06-21T12:59:28Z (GMT). No. of bitstreams: 1 39 DIAS FBS.pdf: 125300 bytes, checksum: 372104899d268ec0b3afbe04e9d6f2fa (MD5) Previous issue date: 2007CNPq, CPqRR-FiocruzFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Triatomíneos e Epidemiologia da Doença de Chagas. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Triatomíneos e Epidemiologia da Doença de Chagas. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Parasitologia Celular e Molecular. Belo Horizonte, MG, BrasilSecretaria do Estado da Saúde do Ceará. Fortaleza, CE, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Triatomíneos e Epidemiologia da Doença de Chagas. Belo Horizonte, MG, BrasilThe aim of this work was to identify and report the occurrence of Trypanosoma rangeli and Trypanosoma cruzi in naturally infected Rhodnius nasutus (Hemiptera, Reduviidae, Triatominae) in the state of Ceará, Brazil. Triatomines feces, salivary glands, and hemolymph were collected for fresh examination, and specific detection of T. rangeli and T. cruzi DNA by polymerase chain reaction was carried out. The specific characterization of these two parasites showed the simultaneous presence of both parasites in two (7.7%) of the 26 positive insects. Our results provide further knowledge on the geographical distribution of T. rangeli in Brazil

Arylfurans as potential Trypanosoma cruzi trypanothione reductase inhibitors

Submitted by Nuzia Santos ([email protected]) on 2013-06-06T18:32:43Z No. of bitstreams: 1 87.pdf: 480919 bytes, checksum: 0ec2d7dcbb146ddb70e377368034b213 (MD5)Made available in DSpace on 2013-06-06T18:32:43Z (GMT). No. of bitstreams: 1 87.pdf: 480919 bytes, checksum: 0ec2d7dcbb146ddb70e377368034b213 (MD5) Previous issue date: 2006Fiocruz, CNPq, FapemigFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Química de Produtos Naturais. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Química de Produtos Naturais. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Parasitologia Celular e Molecular. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Parasitologia Celular e Molecular. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Exatas. Departamento de Química. NEQUIM. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Parasitologia Celular e Molecular. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Química de Produtos Naturais. Belo Horizonte, MG, Brasil.The natural lignans veraguensin and grandisin have been reported to be active against Trypanosoma cruzi bloodstream forms. Aiming at the total synthesis of these and related compounds, we prepared three 2-arylfurans and eight 2,5-diarylfurans. They were evaluated for their potential as T. cruzitrypanothione reductase (TR) inhibi-tors as well against the parasite’s intracellular (amastigote) and bloodstream (trypomastigote) forms. Compound 12 was the most effective against TR with an IC50 of 48.5 µM while 7and 14 were active against amastigotes, inhibiting the parasite development by 60% at 20 µg/ml (59 and 90 µM, respectively). On the other hand, none of the compounds was significantly active against the parasite bloodstream forms even at 250 µg/ml (0.6-1.5 mM)

Experimental Trypanosoma cruzi infection in platelet-activating factor receptor-deficient mice.

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2015-04-08T13:58:03Z No. of bitstreams: 1 Talvani A Experimental trypanosoma....pdf: 165677 bytes, checksum: 67a2e20b99a4c197e86ef3f2c094ec8a (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2015-04-08T14:15:28Z (GMT) No. of bitstreams: 1 Talvani A Experimental trypanosoma....pdf: 165677 bytes, checksum: 67a2e20b99a4c197e86ef3f2c094ec8a (MD5)Made available in DSpace on 2015-04-08T14:15:28Z (GMT). No. of bitstreams: 1 Talvani A Experimental trypanosoma....pdf: 165677 bytes, checksum: 67a2e20b99a4c197e86ef3f2c094ec8a (MD5) Previous issue date: 2003Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, BrasilUniversity of Tokyo. Faculty of Medicine. Department of Biochemistry and Molecular Biology. Tokyo, Japan / CREST of Japan Science and Technology Corporation. Tokyo, JapanUniversity of Tokyo. Faculty of Medicine. Department of Biochemistry and Molecular Biology. Tokyo, Japan / CREST of Japan Science and Technology Corporation. Tokyo, JapanFundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Belo Horizonte, MG, BrasilUniversidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Ribeirão Preto, SP, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Belo Horizonte, MG, BrasilThe generation of an inflammatory response driven by Trypanosoma cruzi or its subproducts appears to be essential for tissue injury and disease pathogenesis. However, this inflammatory response is also relevant in the control of T. cruzi replication. The lipid mediator platelet-activating factor (PAF) has been implicated in a number of pathological conditions characterized by tissue inflammation. In the present study, we aimed at evaluating the role of PAF during T. cruzi infection by using mice that were genetically deficient in the PAF receptor. We observed that infected hearts of PAFR(-/-) mice had an increased number of parasite nests, associated with a more intense inflammatory infiltrate. This was associated with greater parasitemia and lethality. When wild-type and PAFR(-/-) mice were compared, there were no marked changes in the kinetics of the expression of MCP-1, RANTES, IFN-gamma and TNF-alpha in heart tissue of infected animals. Moreover, serum concentrations of TNF-alpha, nitrate and parasite-specific IgM were similar in both groups of mice. In vitro, macrophages from PAFR(-/-) animals did not phagocytose trypomastigote forms when activated with PAF, leukotriene B(4) or MCP-1 and produced less nitric oxide when infected and activated with IFN-gamma. These results are consistent with the hypothesis that endogenous synthesis of PAF and activation of PAF receptors control T. cruzi replication in mice in great part via facilitation of the uptake of the parasite and consequent activation of macrophage