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Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)
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Silva, Andressa Noronha Barbosa da

; Souza, Rita de Cássia Moreira de

; Honorato, Nathan Ravi Medeiros

; Martins, Rand Randall

; Câmara, Antônia Claudia Jácome da

; Galvão, Lúcia Maria da Cunha

; Chiari, Egler

Revista da Sociedade Brasileira de Medicina Tropical

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2020, Volume 53 elocation e20200189

Abstract: En | Text: En | PDF: En

Abstract INTRODUCTION: Knowledge of triatomine bloodmeal sources is essential for understanding vector-host interactions in Trypanosoma cruzi transmission cycles. Expensive commercial deoxyribonucleic acid (DNA) extraction kits are widely used for bloodmeal identification. This study assessed the performance of an inexpensive phenol-chloroform DNA extraction protocol for identification of triatomine bloodmeal sources, comparing it with a commercially available kit. METHODS: Both methods were used to obtain DNA from the intestinal contents of Triatoma brasiliensis blood-fed on either Columba sp., Mus musculus, or Gallus gallus. Subsequently, the mitochondrial 12S ribosomal ribonucleic acid (rRNA) gene was amplified by polymerase chain reaction, sequenced, and compared with GenBank data. RESULTS: Twelve (80%) samples extracted with the commercial kit and four (26.7%) with phenol-chloroform were pure (according to the A260/A280 ratio). Samples extracted with phenol-chloroform, except for Columba sp. samples, had higher DNA concentration than those extracted with the commercial kit. Samples extracted using phenol-chloroform and blood-fed on G. gallus had significantly higher DNA concentration than those blood-fed on Columba sp. (p-value <0.001) and M. musculus (p-value <0.001). The 215-base-pair 12S rRNA fragment was amplified from all samples and produced reliable sequences, enabling the identification of the bloodmeal source, most of which showed identity and coverage above 95%. The phenol-chloroform method was much less expensive than the commercial kit but took considerably more time to perform. CONCLUSIONS: Our data showed that both DNA extraction methods produced reliable sequences enabling identification of triatomine bloodmeal sources but differed greatly in cost and time required.

https://doi.org/10.1590/0037-8682-0189-2020 432 downloads

Synanthropic triatomines (Hemiptera: Reduviidae): infestation, colonization, and natural infection by trypanosomatids in the State of Rio Grande do Norte, Brazil
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Barbosa-Silva, Andressa Noronha

; Souza, Rita de Cássia Moreira de

; Diotaiuti, Liléia

; Aguiar, Lúcia Maria Abrantes

; Câmara, Antonia Cláudia Jácome da

; Galvão, Lúcia Maria da Cunha

; Chiari, Egler

Revista da Sociedade Brasileira de Medicina Tropical

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2019, Volume 52 elocation e20190061

Abstract: En | Text: En | PDF: En

Abstract INTRODUCTION The ecoepidemiological situation in the State of Rio Grande do Norte, Brazil is characterized by frequent invasion and colonization of domiciliary units (DUs) by several triatomine species, with high rates of natural infection by Trypanosoma cruzi. METHODS: We evaluated the possibility of vector transmission of T. cruzi based on records of the occurrence of domiciled triatomines collected by the Secretariat of State for Public Health from 2005 to 2015. During this period, 67.7% (113/167) of municipalities conducted at least one active search and 110 recorded the presence of insects in DUs. These activities were more frequent in municipalities considered to have a high and medium-level risk of T. cruzi transmission. RESULTS Of 51,569 captured triatomines, the most common species were Triatoma brasiliensis (47.2%) and T. pseudomaculata (40.2%). Colonies of T. brasiliensis, T. pseudomaculata, T. petrocchiae, Panstrongylus lutzi, and Rhodnius nasutus were also recorded in the intradomicile and peridomicile. Natural infection by trypanosomatids was detected in 1,153 specimens; the highest rate was found in R. nasutus (3.5%), followed by T. brasiliensis (2.5%) and T. pseudomaculata (2.4%). There have been high levels of colonization over the years; however, not all infested DUs have been sprayed. CONCLUSIONS: This is the first report of intradomicile and peridomicile colonization by P. lutzi. These results demonstrate the risk of new cases of infection by T. cruzi and reinforce the need for continuous entomological surveillance in the State of Rio Grande do Norte.

https://doi.org/10.1590/0037-8682-0061-2019 1229 downloads

Combined parasitological and molecular-based diagnostic tools improve the detection of Trypanosoma cruzi in single peripheral blood samples from patients with Chagas disease
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Volpato, Fabiana Caroline Zempulski

; Sousa, Giovane Rodrigo

; D’Ávila, Daniella Alchaar

; Galvão, Lúcia Maria da Cunha

; Chiari, Egler

Revista da Sociedade Brasileira de Medicina Tropical

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aug 2017, Volume 50 N. 4 Pages 506 - 515

Abstract: En | Text: En | PDF: En

Abstract INTRODUCTION In order to detect Trypanosoma cruzi and determine the genetic profiles of the parasite during the chronic phase of Chagas disease (ChD), parasitological and molecular diagnostic methods were used to assess the blood of 91 patients without specific prior treatment. METHODS Blood samples were collected from 68 patients with cardiac ChD and 23 patients with an indeterminate form of ChD, followed by evaluation using blood culture and polymerase chain reaction. T . cruzi isolates were genotyped using three different genetic markers. RESULTS: Blood culture was positive in 54.9% of all patients, among which 60.3% had the cardiac form of ChD, and 39.1% the indeterminate form of ChD. There were no significant differences in blood culture positivity among patients with cardiac and indeterminate forms. Additionally, patient age and clinical forms did not influence blood culture results. Polymerase chain reaction (PCR) was positive in 98.9% of patients, although comparisons between blood culture and PCR results showed that the two techniques did not agree. Forty-two T . cruzi stocks were isolated, and TcII was detected in 95.2% of isolates. Additionally, one isolate corresponded to TcIII or TcIV, and another corresponded to TcV or TcVI. CONCLUSIONS Blood culture and PCR were both effective for identifying T. cruzi using a single blood sample, and their association did not improve parasite detection. However, we were not able to establish an association between the clinical form of ChD and the genetic profile of the parasite.

https://doi.org/10.1590/0037-8682-0046-2017 1185 downloads

2 nd Brazilian Consensus on Chagas Disease, 2015
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Dias, João Carlos Pinto

; Ramos Jr., Alberto Novaes

; Gontijo, Eliane Dias

; Luquetti, Alejandro

; Shikanai-Yasuda, Maria Aparecida

; Coura, José Rodrigues

; Torres, Rosália Morais

; Melo, José Renan da Cunha

; Almeida, Eros Antonio de

; Oliveira Jr., Wilson de

; Silveira, Antônio Carlos

; Rezende, Joffre Marcondes de

; Pinto, Fabiane Scalabrini

; Ferreira, Antonio Walter

; Rassi, Anis

; Fragata Filho, Abílio Augusto

; Sousa, Andréa Silvestre de

; Correia, Dalmo

; Jansen, Ana Maria

; Andrade, Glaucia Manzan Queiroz

; Britto, Constança Felícia De Paoli de Carvalho

; Pinto, Ana Yecê das Neves

; Rassi Jr., Anis

; Campos, Dayse Elisabeth

; Abad-Franch, Fernando

; Santos, Silvana Eloi

; Chiari, Egler

; Hasslocher-Moreno, Alejandro Marcel

; Moreira, Eliane Furtado

; Marques, Divina Seila de Oliveira

; Silva, Eliane Lages

; Marin-Neto, José Antonio

; Galvão, Lúcia Maria da Cunha

; Xavier, Sergio Salles

; Valente, Sebastião Aldo da Silva

; Carvalho, Noêmia Barbosa

; Cardoso, Alessandra Viana

; Silva, Rafaella Albuquerque e

; Costa, Veruska Maia da

; Vivaldini, Simone Monzani

; Oliveira, Suelene Mamede

; Valente, Vera da Costa

; Lima, Mayara Maia

; Alves, Renato Vieira

Revista da Sociedade Brasileira de Medicina Tropical

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dec 2016, Volume 49 Suppl. 1 Pages 3 - 60

Abstract: En | Text: En | PDF: En

Abstract Chagas disease is a neglected chronic condition with a high burden of morbidity and mortality. It has considerable psychological, social, and economic impacts. The disease represents a significant public health issue in Brazil, with different regional patterns. This document presents the evidence that resulted in the Brazilian Consensus on Chagas Disease. The objective was to review and standardize strategies for diagnosis, treatment, prevention, and control of Chagas disease in the country, based on the available scientific evidence. The consensus is based on the articulation and strategic contribution of renowned Brazilian experts with knowledge and experience on various aspects of the disease. It is the result of a close collaboration between the Brazilian Society of Tropical Medicine and the Ministry of Health. It is hoped that this document will strengthen the development of integrated actions against Chagas disease in the country, focusing on epidemiology, management, comprehensive care (including families and communities), communication, information, education, and research .

https://doi.org/10.1590/0037-8682-0505-2016 9267 downloads

II Consenso Brasileiro em Doença de Chagas, 2015
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Dias, João Carlos Pinto

; Ramos Jr., Alberto Novaes

; Gontijo, Eliane Dias

; Luquetti, Alejandro

; Shikanai-Yasuda, Maria Aparecida

; Coura, José Rodrigues

; Torres, Rosália Morais

; Melo, José Renan da Cunha

; Almeida, Eros Antonio de

; Oliveira Jr., Wilson de

; Silveira, Antônio Carlos

; Rezende, Joffre Marcondes de

; Pinto, Fabiane Scalabrini

; Ferreira, Antonio Walter

; Rassi, Anis

; Fragata Filho, Abílio Augusto

; Sousa, Andréa Silvestre de

; Correia Filho, Dalmo

; Jansen, Ana Maria

; Andrade, Glaucia Manzan Queiroz

; Britto, Constança Felícia De Paoli de Carvalho

; Pinto, Ana Yecê das Neves

; Rassi Jr., Anis

; Campos, Dayse Elisabeth

; Abad-Franch, Fernando

; Santos, Silvana Eloi

; Chiari, Egler

; Hasslocher-Moreno, Alejandro Marcel

; Moreira, Eliane Furtado

; Marques, Divina Seila de Oliveira

; Silva, Eliane Lages

; Marin-Neto, José Antonio

; Galvão, Lúcia Maria da Cunha

; Xavier, Sergio Salles

; Valente, Sebastião Aldo da Silva

; Carvalho, Noêmia Barbosa

; Cardoso, Alessandra Viana

; Silva, Rafaella Albuquerque e

; Costa, Veruska Maia da

; Vivaldini, Simone Monzani

; Oliveira, Suelene Mamede

; Valente, Vera da Costa

; Lima, Mayara Maia

; Alves, Renato Vieira

Epidemiologia e Serviços de Saúde

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jun 2016, Volume 25 N. spe. Pages 7 - 86

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO A doença de Chagas é uma condição crônica negligenciada com elevada carga de morbimortalidade e impacto dos pontos de vista psicológico, social e econômico. Representa um importante problema de saúde pública no Brasil, com diferentes cenários regionais. Este documento traduz a sistematização das evidências que compõe o Consenso Brasileiro de Doença de Chagas. O objetivo foi sistematizar estratégias de diagnóstico, tratamento, prevenção e controle da doença de Chagas no país, de modo a refletir as evidências científicas disponíveis. Sua construção fundamentou-se na articulação e contribuição estratégica de especialistas brasileiros com conhecimento, experiência e atualização sobre diferentes aspectos da doença. Representa o resultado da estreita colaboração entre a Sociedade Brasileira de Medicina Tropical e o Ministério da Saúde. Espera-se com este documento fortalecer o desenvolvimento de ações integradas para enfrentamento da doença no país com foco em epidemiologia, gestão, atenção integral (incluindo famílias e comunidades), comunicação, informação, educação e pesquisas.

ABSTRACT Chagas disease is a neglected chronic condition that presents high morbidity and mortality burden, with considerable psychological, social, and economic impact. The disease represents a significant public health issue in Brazil, with different regional patterns. This document presents the evidence that resulted in the Brazilian Consensus on Chagas Disease. The objective was to review and standardize strategies for diagnosis, treatment, prevention, and control of Chagas disease in the country, based on the available scientific evidence. The consensus is based on collaboration and contribution of renowned Brazilian experts with vast knowledge and experience on various aspects of the disease. It is the result of close collaboration between the Brazilian Society of Tropical Medicine and the Ministry of Health. This document shall strengthen the development of integrated control measures against Chagas disease in the country, focusing on epidemiology, management, comprehensive care (including families and communities), communication, information, education, and research.

https://doi.org/10.5123/S1679-49742016000500002 3725 downloads

II Consenso Brasileiro em Doença de Chagas, 2015
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Dias, João Carlos Pinto

; Ramos Jr., Alberto Novaes

; Gontijo, Eliane Dias

; Luquetti, Alejandro

; Shikanai-Yasuda, Maria Aparecida

; Coura, José Rodrigues

; Torres, Rosália Morais

; Melo, José Renan da Cunha

; Almeida, Eros Antonio de

; Oliveira Jr., Wilson de

; Silveira, Antônio Carlos

; Rezende, Joffre Marcondes de

; Pinto, Fabiane Scalabrini

; Ferreira, Antonio Walter

; Rassi, Anis

; Fragata Filho, Abílio Augusto

; Sousa, Andréa Silvestre de

; Correia Filho, Dalmo

; Jansen, Ana Maria

; Andrade, Glaucia Manzan Queiroz

; Britto, Constança Felícia De Paoli de Carvalho

; Pinto, Ana Yecê das Neves

; Rassi Jr., Anis

; Campos, Dayse Elisabeth

; Abad-Franch, Fernando

; Santos, Silvana Eloi

; Chiari, Egler

; Hasslocher-Moreno, Alejandro Marcel

; Moreira, Eliane Furtado

; Marques, Divina Seila de Oliveira

; Silva, Eliane Lages

; Marin-Neto, José Antonio

; Galvão, Lúcia Maria da Cunha

; Xavier, Sergio Salles

; Valente, Sebastião Aldo da Silva

; Carvalho, Noêmia Barbosa

; Cardoso, Alessandra Viana

; Silva, Rafaella Albuquerque e

; Costa, Veruska Maia da

; Vivaldini, Simone Monzani

; Oliveira, Suelene Mamede

; Valente, Vera da Costa

; Lima, Mayara Maia

; Alves, Renato Vieira

Epidemiologia e Serviços de Saúde

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jun 2016, Volume 25 N. spe Pages 7 - 86

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

https://doi.org/10.5123/s1679-49742016000500002 63592 downloads

Characteristics of Triatomine infestation and natural Trypanosoma cruzi infection in the State of Rio Grande do Norte, Brazil
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Barbosa-Silva, Andressa Noronha

; Câmara, Antonia Cláudia Jácome da

; Martins, Kiev

; Nunes, Daniela Ferreira

; Oliveira, Pedro Igor Câmara de

; Azevedo, Paulo Roberto Medeiros de

; Chiari, Egler

; Galvão, Lúcia Maria da Cunha

Revista da Sociedade Brasileira de Medicina Tropical

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feb 2016, Volume 49 N. 1 Pages 57 - 67

Abstract: En | Text: En | PDF: En

Abstract: INTRODUCTION Natural and artificial ecotope infestation by the kissing bug triatomines and their colonization and infection by Trypanosoma cruzi , the Chagas disease agent, were evaluated in nine municipalities of the State of Rio Grande do Norte, Brazil. METHODS Following identification, triatomine intestinal contents were analyzed by direct microscopic examination, xenoculture, and polymerase chain reaction (PCR) for parasite detection. Trypanosoma cruzi isolates were genotyped using three different markers. RESULTS Of 842 triatomines captured, 65% were Triatoma brasiliensis , 17.8% Triatoma pseudomaculata , 12.5% Panstrongylus lutzi , and 4.7% Rhodnius nasutus . Triatoma brasiliensis and P. lutzi adults were found in the intradomicile. T. brasiliensis, T. pseudomaculata , and R. nasutus nymphs and adults were found in the peridomicile and wild environment. Intradomiciliary and peridomiciliary infestation indexes were 5.6% and 33.7%, respectively. In the peridomicile, chicken coops were the most infested ecotope. The T. cruzi triatomine infection rate was 30.2%, of which PCR detected 29%. P . lutzi (78.1%), T . brasiliensis (24.5%), and T . pseudomaculata (22.7%) were the most infected species. TcII and III genotypes were detected in T. brasiliensis and TcIII in P. lutzi . CONCLUSIONS T. brasiliensis was found in all environments and most ecotopes with high T. cruzi infection rates. High infection rates were also detected in T . pseudomaculata and P. lutzi , suggesting their role in the interchange between the wild and peridomestic transmission cycles. The combination of PCR, microscopic examination, and xenoculture contributed to improving T. cruzi infection evaluation in triatomine bugs. The TcII and TcIII genotypes were predominant in the study area.

https://doi.org/10.1590/0037-8682-0300-2015 2569 downloads

Chagas disease: morbidity profile in an endemic area of Northeastern Brazil
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Andrade, Cléber de Mesquita

; Câmara, Antônia Cláudia Jácome da

; Nunes, Daniela Ferreira

; Guedes, Paulo Marcos da Matta

; Pereira, Wogelsanger Oliveira

; Chiari, Egler

; Diniz, Rosiane Viana Zuza

; Galvão, Lúcia Maria da Cunha

Revista da Sociedade Brasileira de Medicina Tropical

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dec 2015, Volume 48 N. 6 Pages 706 - 715

Abstract: En | Text: En | PDF: En

Abstract: INTRODUCTION : This study evaluated the clinical forms and manifestation severities of Chagas disease among serologically reactive individuals from Western Rio Grande do Norte (Northeastern Brazil). METHODS : This cross-sectional study included 186 adults who were evaluated using electrocardiography, echocardiography, chest radiography, and contrast radiography of the esophagus and colon. A clinical-epidemiological questionnaire was also used. RESULTS : The indeterminate, cardiac, digestive, and cardiodigestive clinical forms of Chagas disease were diagnosed in 51.6% (96/186), 32.2% (60/186), 8.1% (15/186) and 8.1% (15/186) of the participants, respectively. Heart failure (functional classes I-IV) was detected in 7.5% (14/186) of the participants, and 36.4% (24/66), 30.3% (20/66), 15.2% (10/66), 13.6% (9/66), and 4.5% (3/66) of the patients were at stage A, B1, B2, C, and D, respectively. Dilated cardiomyopathy and electrocardiographic changes were detected in 10.2% (19/186) and 48.1% (91/186) of the participants, respectively. Apical aneurysm was diagnosed in 10.8% (20/186) of the participants, and other changes in the segmental myocardial contractility of the left ventricle were diagnosed in 33.9% (63/186) of the participants. Megaesophagus (groups I-IV) was observed in 7% (13/186) of the participants, megacolon (grades 1-3) was detected in12.9% (24/186) of the participants, and both organs were affected in 29.2% (7/24) of the megacolon cases. CONCLUSIONS : We detected various clinical forms of Chagas disease (including the digestive form). Our findings indicate that clinical symptoms alone may not be sufficient to exclude or confirm cardiac and/or digestive damage, and the number of patients with symptomatic clinical forms may be underestimated.

https://doi.org/10.1590/0037-8682-0235-2015 3188 downloads

Complete assignments of NMR data and assessment of trypanocidal activity of new eremantholide C derivatives
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SAÚDE-GUIMARÃES, DÊNIA A.

; RASLAN, DÉLIO S.

; CHIARI, EGLER

; OLIVEIRA, ALAÍDE B. DE

Anais da Academia Brasileira de Ciências

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dec 2014, Volume 86 N. 4 Pages 1563 - 1572

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Transformações químicas realizadas em eremantolida C (1), uma lactona sesquiterpênica isolada de Lychnophora trichocarpha Spreng. originaram cinco novos derivados: 1′,2′-epoxieremantolida C (2), 5-n-propilamino-4,5-diidro-1′,2′-epoxieremantolida C (3), cloreto de 5-n-propilamônio-4,5-diidro-1′, 2′-epoxieremantolida C (4), cloreto de 5-n-propilamônio-4,5-diidroeremantolida C (5) e 16-O-etileremantolida C (6). As estruturas químicas de todos estes derivados foram elucidadas com base nos espectros de IV, EM, RMN de 1H e de 13C e por meio de técnicas 1D e 2D. Eremantolida C e os derivados 2, 4 e 5 foram avaliados frente a cepas tripomastigotas Y e CL de Trypanosoma cruzi. Eremantolida C inibiu completamente o crescimento de ambas as cepas de parasitas, enquanto todos os derivados foram parcialmente ativos contra a cepa CL e inativos contra a cepa Y.

Chemical transformations of eremantholide C (1), a sesquiterpene lactone that was isolated from Lychnophora trichocarpha Spreng. led to five new derivatives: 1′,2′- epoxyeremantholide C (2), 5-n-propylamine-4,5-dihydro-1′,2′-epoxyeremantholide C (3), 5-n-propylammonium-4,5-dihydro-1′,2′-epoxyeremantholide C chloride (4), 5-n-propylammonium-4,5-dihydroeremantolide C chloride (5) and 16-O-ethyleremantholide C (6). The structures of all these derivatives were assigned on the basis of IR, MS, 1H and 13C NMR data by 1D and 2D techniques. Eremantholide C and the derivatives 2, 4 and 5 were evaluated against trypomastigotes Y and CL strains of Trypanosoma cruzi. Eremantholide C completely inhibited the growth of both the parasites strains while all derivatives were partially active against the CL strain and inactive against the Y strain.

https://doi.org/10.1590/0001-3765201420140167 1759 downloads

10.

Hematological alterations during experimental canine infection by Trypanosoma cruzi
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Guedes, Paulo Marcos Matta

; Veloso, Vanja Maria

; Mineo, Tiago Wilson Patriarca

; Santiago-Silva, Juliana

; Crepalde, Geovan

; Caldas, Ivo Santana

; Nascimento, Manuela Sales Lima

; Lana, Marta

; Chiari, Egler

; Galvão, Lúcia Maria da Cunha

; Bahia, Maria Terezinha

Revista Brasileira de Parasitologia Veterinária

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jun 2012, Volume 21 N. 2 Pages 151 - 156

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Para confirmar que cães Beagle são um bom modelo para doença de Chagas, foram avaliadas as alterações hematológicas durante as fases aguda e crônica em cães Beagle infectados com as cepas Y, Berenice-78 (Be-78) e ABC de Trypanosomacruzi, correlacionando os sinais clínicos com a curva de parasitemia. Foi demonstrado que a fase aguda da infecção foi marcada por letargia e perda de apetite. Simultaneamente, observou-se anemia, leucocitose e linfocitose. Ainda, foram descritas alterações hematológicas e sinais clínicos positivamente correlacionados com a parasitemia durante a infecção experimental com as três cepas de T.cruzi estudadas, demonstrando que a infecção em cães Beagle constitui um modelo fidedigno para a doença de Chagas.

To confirm that Beagle dogs are a good experimental model for Chagas disease, we evaluated hematological alterations during the acute and chronic phases in Beagle dogs infected with the Y, Berenice-78 (Be-78) and ABC strains of Trypanosomacruzi, correlating clinical signs with the parasitemia curve. We demonstrate that the acute phase of infection was marked by lethargy and loss of appetite. Simultaneously, we observed anemia, leukocytosis and lymphocytosis. Also,we describe hematological alterations and clinical signs that were positively correlated with the parasitemia during the experimental infection with the three strains of T.cruzi, and demonstrate that experimental infection of Beagle is a trustworthy model for Chagas disease.

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11.

Seroepidemiology of Trypanosoma cruzi infection in the semiarid rural zone of the State of Rio Grande do Norte, Brazil
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Brito, Carlos Ramon do Nascimento

; Sampaio, George Harisson Felinto

; Câmara, Antonia Cláudia Jácome da

; Nunes, Daniela Ferreira

; Azevedo, Paulo Roberto Medeiros de

; Chiari, Egler

; Galvão, Lúcia Maria da Cunha

Revista da Sociedade Brasileira de Medicina Tropical

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jun 2012, Volume 45 N. 3 Pages 346 - 352

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

INTRODUÇÃO: A infecção pelo Trypanosoma cruzi foi avaliada em uma área endêmica do Estado do Rio Grande do Norte, Brasil, por inquérito soroepidemiológico amostral em moradores da zona rural. MÉTODOS: Dezesseis municípios foram sorteados, 15 da mesorregião oeste e um da central, com população estimada em 83.852 indivíduos. Foram coletadas 1.950 amostras de sangue no oeste e 390 em Caicó. A detecção de anticorpos anti-T. cruzi foi realizada usando os kits Chagatest® ELISA, HAI-hemaglutinação e a reação de imunofluorescência indireta. Nos soros com resultados indeterminados foi realizado o western blot TESAcruzi® para confirmação da reatividade. RESULTADOS: A estimativa da soroprevalência revelou 6,5% para a mesorregião oeste e 3,3% em Caicó. A soropositividade eleva-se progressivamente com a idade dos indivíduos até a quinta década de vida em Caicó e na sexta década na mesorregião oeste. Apenas o grau de escolaridade e o conhecimento do triatomíneo evidenciaram associação à soropositividade. Não foram identificados indivíduos sororreativos com idade inferior a 18 anos. CONCLUSÕES: A infecção pelo T. cruzi persiste mais elevada e concentrada em municípios da área central da mesorregião oeste, mas evidências sugerem o declínio da transmissão vetorial nessa mesorregião e em Caicó. As variáveis epidemiológicas parecem não exercer influência na soropositividade, à exceção da escolaridade e conhecimento do triatomíneo entre indivíduos sororreativos da mesorregião oeste.

INTRODUCTION: A seroepidemiological survey was carried out to evaluate Trypanosoma cruzi infection in an endemic area of the State of Rio Grande do Norte, Brazil, involving rural residents. METHODS: Sixteen municipalities were randomly selected, 15 from the west mesoregion and one from the central, with an estimated population of 83,852 individuals. A total of 1,950 blood samples were collected in the west mesoregion and 390 in Caicó. Anti-T. cruzi antibodies were detected using the Chagatest® ELISA HAI-hemagglutination kits and indirect immunofluorescence. As sera presented indeterminate results, TESAcruzi® western blot was performed to confirm reactivity. RESULTS: An estimated seroprevalence of 6.5% was determined for the west mesoregion and 3.3% for Caicó. Seropositivity rises progressively with the age of individuals, up to 40 years in Caicó and up to 50 years in the west mesoregion. Only educational level and knowledge regarding the triatomine were associated with seropositivity. No seroreactive individuals under 18 years of age were identified. CONCLUSIONS: Infection by T. cruzi remains high and is concentrated in municipalities in the central western area of the west mesoregion; however, evidence suggests a decline in vector transmission in this mesoregion and in Caicó. Epidemiological variables appear not to influence seropositivity, with the exception of education and knowledge concerning the triatomine, among seroreactive individuals from the west mesoregion.

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12.

Trypanosoma cruzi benznidazole susceptibility in vitro does not predict the therapeutic outcome of human Chagas disease
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Moreno, Margoth

; D'ávila, Daniella A

; Silva, Marcelo N

; Galvão, Lúcia MC

; Macedo, Andrea M

; Chiari, Egler

; Gontijo, Eliane D

; Zingales, Bianca

Memórias do Instituto Oswaldo Cruz

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nov 2010, Volume 105 N. 7 Pages 918 - 924

Abstract: En | Text: En | PDF: En

Therapeutic failure of benznidazole (BZ) is widely documented in Chagas disease and has been primarily associated with variations in the drug susceptibility of Trypanosoma cruzi strains. In humans, therapeutic success has been assessed by the negativation of anti-T. cruzi antibodies, a process that may take up to 10 years. A protocol for early screening of the drug resistance of infective strains would be valuable for orienting physicians towards alternative therapies, with a combination of existing drugs or new anti-T. cruzi agents. We developed a procedure that couples the isolation of parasites by haemoculture with quantification of BZ susceptibility in the resultant epimastigote forms. BZ activity was standardized with reference strains, which showed IC50 to BZ between 7.6-32 µM. The assay was then applied to isolates from seven chronic patients prior to administration of BZ therapy. The IC50 of the strains varied from 15.6 ± 3-51.4 ± 1 µM. Comparison of BZ susceptibility of the pre-treatment isolates of patients considered cured by several criteria and of non-cured patients indicates that the assay does not predict therapeutic outcome. A two-fold increase in BZ resistance in the post-treatment isolates of two patients was verified. Based on the profile of nine microsatellite loci, sub-population selection in non-cured patients was ruled out.

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13.

Differential tissue tropism of Trypanosoma cruzi strains: an in vitro study
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Andrade, Luciana O

; Galvão, Lúcia MC

; Meirelles, Maria de Nazareth SL

; Chiari, Egler

; Pena, Sergio DJ

; Macedo, Andrea M

Memórias do Instituto Oswaldo Cruz

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sep 2010, Volume 105 N. 6 Pages 834 - 837

Abstract: En | Text: En | PDF: En

We have previously demonstrated selection favoring the JG strain of Trypanosoma cruziin hearts of BALB/c mice that were chronically infected with an equal mixture of the monoclonal JG strain and a clone of the Colombian strain, Col1.7G2. To evaluate whether cell invasion efficiency drives this selection, we infected primary cultures of BALB/c cardiomyocytes using these same T. cruzi populations. Contrary to expectation, Col1.7G2 parasites invaded heart cell cultures in higher numbers than JG parasites; however, intracellular multiplication of JG parasites was more efficient than that of Col1.7G2 parasites. This phenomenon was only observed for cardiomyocytes and not for cultured Vero cells. Double infections (Col1.7G2 + JG) showed similar results. Even though invasion might influence tissue selection, our data strongly suggest that intracellular development is important to determine parasite tissue tropism.

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14.

In vitro and in vivo experimental models for drug screening and development for Chagas disease
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Romanha, Alvaro José

; Castro, Solange Lisboa de

; Soeiro, Maria de Nazaré Correia

; Lannes-Vieira, Joseli

; Ribeiro, Isabela

; Talvani, André

; Bourdin, Bernadette

; Blum, Bethania

; Olivieri, Bianca

; Zani, Carlos

; Spadafora, Carmenza

; Chiari, Egler

; Chatelain, Eric

; Chaves, Gabriela

; Calzada, José Eduardo

; Bustamante, Juan Manuel

; Freitas-Junior, Lucio H

; Romero, Luz I

; Bahia, Maria Terezinha

; Lotrowska, Michel

; Soares, Milena

; Andrade, Sonia Gumes

; Armstrong, Tanya

; Degrave, Wim

; Andrade, Zilton de Araújo

Memórias do Instituto Oswaldo Cruz

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mar 2010, Volume 105 N. 2 Pages 233 - 238

Abstract: En | Text: En | PDF: En

Chagas disease, a neglected illness, affects nearly 12-14 million people in endemic areas of Latin America. Although the occurrence of acute cases sharply has declined due to Southern Cone Initiative efforts to control vector transmission, there still remain serious challenges, including the maintenance of sustainable public policies for Chagas disease control and the urgent need for better drugs to treat chagasic patients. Since the introduction of benznidazole and nifurtimox approximately 40 years ago, many natural and synthetic compounds have been assayed against Trypanosoma cruzi, yet only a few compounds have advanced to clinical trials. This reflects, at least in part, the lack of consensus regarding appropriate in vitro and in vivo screening protocols as well as the lack of biomarkers for treating parasitaemia. The development of more effective drugs requires (i) the identification and validation of parasite targets, (ii) compounds to be screened against the targets or the whole parasite and (iii) a panel of minimum standardised procedures to advance leading compounds to clinical trials. This third aim was the topic of the workshop entitled Experimental Models in Drug Screening and Development for Chagas Disease, held in Rio de Janeiro, Brazil, on the 25th and 26th of November 2008 by the Fiocruz Program for Research and Technological Development on Chagas Disease and Drugs for Neglected Diseases Initiative. During the meeting, the minimum steps, requirements and decision gates for the determination of the efficacy of novel drugs for T. cruzi control were evaluated by interdisciplinary experts and an in vitro and in vivo flowchart was designed to serve as a general and standardised protocol for screening potential drugs for the treatment of Chagas disease.

12866 downloads Cited 9 times in SciELO

15.

Immunological imbalance between IFN-³ and IL-10 levels in the sera of patients with the cardiac form of Chagas disease
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D'Ávila, Daniela A

; Guedes, Paulo MM

; Castro, Ana M

; Gontijo, Eliane D

; Chiari, Egler

; Galvão, Lúcia MC

Memórias do Instituto Oswaldo Cruz

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feb 2009, Volume 104 N. 1 Pages 100 - 105

Abstract: En | Text: En | PDF: En

The immune response is crucial for protection against disease; however, immunological imbalances can lead to heart and digestive tract lesions in chagasic patients. Several studies have evaluated the cellular and humoral immune responses in chagasic patients in an attempt to correlate immunological findings with clinical forms of Chagas disease. Moreover, immunoglobulins and cytokines are important for parasitic control and are involved in lesion genesis. Here, cytokine and IgG isotype production were studied, using total epimastigote antigen on sera of chagasic patients with indeterminate (IND, n = 27) and cardiac (CARD, n = 16) forms of the disease. Samples from normal, uninfected individuals (NI, n = 30) were use as controls. The results showed that sera from both IND and CARD patients contained higher levels of Trypanosoma cruzi-specific IgG1 (IgG1) antibodies than sera from NI. No difference in IgG2 production levels was observed between NI, IND and CARD patients, nor was a difference in IL-10 and IFN-³ production detected in the sera of IND, CARD and NI patients. However, IND patients displayed a positive correlation between IL-10 and IFN-³ levels in serum, while CARD patients showed no such correlation, indicating an uncontrolled inflammatory response in CARD patients. These findings support the hypothesis that a lack of efficient regulation between IFN-³ and IL-10 productions in CARD patients may lead to cardiac immunopathology.

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