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Asian Pacific Journal of Tropical Medicine 2014年第11期7卷 854-859页

作者： Francisco Olmo Javier Escobedo-Ortegón Patricia Palma Manuel Sánchez-Moreno Ana Mejía-Jaramillo Omar Triana Clotilde Marín Grupo de Parasitologia Molecular Departmento de Parasitologia Universidad de Granada Laboratorio de Zoonosis y ETV's C.I.R.Dr.Hideyo NoguchiUniversidad Autonoma de YucatanAve.Itzaes #490 x 5997000 MeridaYucatan(Mexico) Grupo Biologiay Control de Enfermedades Infecciosas BCEI-SIU Instituto de BiologiaUniversidad de Antioquia

Objective:To classify 21 new isolates of Trypanosoma cruai(T.cruzi) according to the Discrete Typing Unit(DTU) which they belong to,as well as tune up a new pair of primers designed to detect the parasite in biological samples.Methods:Strains were isolated,DNA extracted,and classified by using three Polymerase Chain Reactions(PCR).Subsequently this DNA was used along with other isolates of various biological samples,for a new PCR using primers designed.Finally,the amplified fragments were sequenced.Results:It was observed the predominance of DTU i in Colombia,as well as the specificity of our primers for detection of T.cruzi,while no band was obtained when other species were used.Conclusions:This work reveals the genetic variability of 21 new isolates of T.cruzi in Colombia.Our primers confirmed their specificity for detecting the presence of T.cruzi.

关键词： Trypanosoma cruzi Polymerase chain reaction Colombia Superoxide dismutase gene b-based primers Discrete typing unit

Canova medication changes TNF-α and IL-10 serum levels in mice infected with Trypanosoma cruzi Y strain

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Asian Pacific Journal of Tropical Medicine 2016年第9期9卷 838-843页

作者： áurea Regina Telles Pupulin Lívia Bracht Márcia Machado de Oliveira Dalalio Francielli Maria de Souza Silva-Comar Bruno Ambrósio da Rocha Franciele Queiroz Ames Roberto Kenji Nakamura Cuman Ciomar Aparecida Bersani-Amado Laboratory of Parasitology Department of Basic Sciences of HealthState University of Maringá-PR Laboratory of Hepatic Metabolism Department of BiochemistryState University of Maringá-PR Laboratory of Immunology Department of Basic Sciences of HealthState University of Maringá-PR Laboratory of Inflammation Department of Pharmacology and TherapeuticState University of Maringá-PR

Objective:To identify whether Canova medication changes TNF-α and IL-10 serum levels in mice infected with Trypanosoma cruzi Y strain.Methods:Animals were divided into five groups:non-treated infected animals(I); benznidazole-treated infected animals(Bz; 100 mg/kg body weight,single daily dose by gavage); Canova medication(CM) treated infected animals(CM;0.2 mL/animal,single daily dose by gavage); benznidazole- and Canova medication–treated infected animals with the above-mentioned dose(Bz+CM);and non-infected animals(C).TNF-α and IL-10 levels were determined in serum aliquots after 4,7,10,13,and 29 days of infection.An ELISA technique was employed with R&D System Inc.antibody pairs.Results:A high increase in TNF-α and IL-10 levels occurred in the infected and CM-treated groups within the treatment employed on the 10 th day after infection,coupled with a IL-10 decrease on the 13 th day after infection when compared with the other experimental groups.Conclusions:CM may change the balance between plasma cytokine levels(TNF-α and IL-10) in mice infected with Y strain T.cruzi,with important consequences leading towards a more severe infection.

关键词： Trypanosoma cruzi Canova medication Cytokines

Antitrypanosomal Activity of a Semi-Purified Subfraction Rich in Labdane Sesquiterpenes, Obtained from Flowers of Anthemis Tinctoria, Against Trypanosoma cruzi

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Pharmacology & Pharmacy 2011年第2期2卷 47-55页

作者： Nilza de Lucas Rodrigues Bittencourt Tania Ueda-Nakamura Benedito Prado Dias Filho Celso Vataru Nakamura 不详

In Brazil and several other Latin American countries, Chagas' disease still constitutes a serious medical and social problem, and there is a need to develop new, more-potent drugs with fewer side effects to effectively treat this disease. We investigated the antitrypanosomal effect of a crude extract, fractions, and a semi-purified subfraction rich in a mix- ture of isomeric labdane sesquiterpenes, obtained from flowers of Anthemis tinctoria, against Trypanosoma cruzi. In epimastigote forms, the aqueous crude extract, dichloromethane fraction, and semi-purified subfraction showed a dose-dependent inhibitory activity, with IC50 of 2.3 μg/ml, 1.8 μg/ml, and 0.2 μg/ml, respectively. In the interaction in- dex, the semi-purified subfraction showed a reduction in both the percentage of infected LLCMK2 cells and the mean number of amastigotes per infected cell. The cytotoxicity evaluation demonstrated that the cytotoxic concentrations of the semi-purified subfraction were higher for LLCMK2 cells than for the protozoans, with a selectivity index of 35.0. Epimastigote forms treated with the semi-purified subfraction showed ultrastructural and morphological alterations such as rounding of the cells and bleb formation in the flagellum and cytoplasmic membrane. These results show that the flowers from A. tinctoria may be a source of new drugs with antiprotozoal activity. However, additional in vitro and in vivo studies are needed to validate the use of A. tinctoria in the treatment of Chagas’ disease.

关键词： Antiprotozoan Activity Medicinal Plants Trypanosoma cruzi Ultrastructure Alterations

Antibody delivery into viable epimastigotes of <i>Trypanosoma cruzi</i>as a tool to study the parasite biology

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Advances in Bioscience and Biotechnology 2013年第6期4卷 719-726页

作者： Karla Y. Acosta-Viana Huchin-Cetz Julio Jimenez-Coello Matilde Guzman-Marin Eugenia Jose L. Rosales-Encina Departamento de Infectomica y Patogenesis Molecular Centro de Investigación y de Estudios Avanzados del Instituto Politecnico Nacional Unidad Zacatenco México Laboratorio Biologia Celular Centro de Investigaciones Regionales “Dr. Hideyo Noguchi” Universidad Autonoma de Yucatan Merida Mexico

American trypanosomiasis is a zoonosis of worldwide medical importance and currently there is no effective treatment in chronic patients, hence the importance of the study of protein function of the parasite with the objective of finding new drug targets and to know better the biology of the agent causal (Trypano-soma cruzi). T. cruzi is an RNAi-negative parasite, therefore the silencing genes strategies by RNAi is not possible;for that reason, antibodies may be taken as a tool for studying the parasite proteins function by blocking these molecules with specific antibodies. The aim of this work was to establish a methodology for antibody delivery (antibody transfection) into viable parasites. We used anti-cyclin-A antibody (human origin) in western blot assay with epimastigote of T. cruzi proteins and this recognized a ~55 kDa polypeptide. Several methods for antibody transfection (electroporation, saponin permeabilization and a lipid-based formulation) were tested. The first two methods were unsuccessful. In electroporation was impossible to visualize the antibody inside parasites and with saponin permeabilization, antibodies were successfully introduced, but with loss of parasites viability. The lipid-based formulation method forms noncovalent complexes with antibodies. These complexes are internalized by cells and antibodies are released into the cytoplasm. With this method, a successful antibody delivery was achieved. Anti-cyclin antibodies were visualized in the cytoplasm from fixed transfected parasites (immunofluorescence assays). At 24 h post-transfection, parasites maintained their viability (90%) and were able to arrest the cell cycle in G0/G1-phase of cultured epimastigotes (cell population increased in G0/G1-phase from 50.5% to 66.2% and decreased in S-phase from 47.2% to 26%). It was also observed that anti-cyclin-A antibodies inhibit the parasite population doubling (p T. cruzi, with a simple and cheap technique, which will allows carrying out further studies of this protozoan.

关键词： Trypanosoma cruzi Antibody Delivery Antibody Transfection Cell Cycle Anti-Cyclin-A Antibody

The association of Triatoma maculata(Ericsson 1848) with the gecko Thecadactylus rapicauda(Houttuyn 1782)(Reptilia:Squamata:Gekkonidae):a strategy of domiciliation of the Chagas disease peridomestic vector in Venezuela?

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Asian Pacific Journal of Tropical Biomedicine 2011年第4期1卷 279-284页

作者： Reyes-Lugo M Reyes-Contreras M Salvi I Gelves W Avilan A Llavaneras D Navarrete LF Cordero G Sanchez EE Rodriguez-Acosta A Medical Entomology Section Institute of Tropical MedicineUniversidad Central de VenezuelaCaracasVenezuela School of Biology Universidad Simon BolivarCaracasVenezuela Immunochemistry Section Tropical Medicine InstituteUniversidad Central de VenezuelaCaracasVenezuela Tropical Zoology Institute Universidad Central de VenezuelaCaracasVenezuela Texas A&M University-Kingsville Department of ChemistryKingsvilleTexasUSA Instituto Nacional de Higiene Rafael Rangel CaracasVenezuela

Objective:To investigate the bioecological relationship between Chagas disease peridomestic vectors and reptiles as source of feeding.Methods:In a three-story building,triatomines were captured by direct search and electric vacuum cleaner search in and outside the building.Then,age structure of the captured Triatoma maculata(T.maculata) were identified and recorded.Reptiles living in sympatric with the triatomines were also searched.Results:T.maculata were found living sympatric with geckos(Thecadactylus rapicauda) and they bit residents of the apartment building in study.A total of 1 448 individuals of T.maculata were captured within three days,of which 74.2%(1 074 eggs) were eggs,21.5%were nymphs at different stages,and 4.3%were adults.Conclusions:The association of T.maculata and T.rapicauda is an effective strategy of colonizing dwellings located in the vicinity of the habitat where both species are present;and therefore,could have implications of high importance in the intradomiciliary transmission of Chagas disease.

关键词： Chagas disease Gekkonidae Reduviidae Thecadactylus rapicauda Triatoma maculate Trypanosoma cruzi Vector domiciliation

Modulation of immune response in experimental Chagas disease

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World Journal of Experimental Medicine 2013年第1期3卷 1-10页

作者： Beatriz Basso Neonatology Service School of Medical SciencesNational University of Cordoba

Trypanosoma cruzi(T. cruzi), the etiological agent of Chagas disease, affects nearly 18 million people in Latin America and 90 million are at risk of infection. The parasite presents two stages of medical importance in the host, the amastigote, intracellular replicating form, and the extracellular trypomastigote, the infective form. Thus infection by T. cruzi induces a complex immune response that involves effectors and regulatory mechanisms. That is why control of the infection requires a strong humoral and cellular immune response; hence, the outcome of host-parasite interaction in the early stages of infection is extremely important. A critical event during this period of the infection is innate immune response, in which the macrophage's role is vital. Thus, after being phagocytized, the parasite is able to develop intracellularly; however, during later periods, these cells induce its elimination by means of toxic metabolites. In turn, as the infection progresses, adaptive immune response mechanisms are triggered through the TH1 and TH2 responses. Finally, T. cruzi, like other protozoa such as Leishmania and Toxoplasma, have numerous evasive mechanisms to the immune response that make it possible to spread around the host. In our Laboratory we have developed a vaccination model in mice with Trypanosoma rangeli, nonpathogenic to humans, which modulates the immune response to infection by T. cruzi, thus protecting them. Vaccinated animals showed an important innate response(modulation of NO and other metabolites, cytokines, activation of macrophages), a strong adaptive cellular response and significant increase in specific antibodies. The modulation caused early elimination of the parasites, low parasitaemia, the absence of histological lesions and high survival rates. Even though progress has been made in the knowledge of some of these mechanisms, new studies must be conducted which could target further prophylactic and therapeutic trials against T. cruzi infection.

关键词： Trypanosoma cruzi Chagas disease Innate and adaptive immune response

Synthesis and Trypanocidal Evaluation of Some Novel 2-(Substituted Benzylidene)-5, 7-Dibromo-6-Hydroxy-1-Benzofuran-3(2<i>H</i>)-Ones

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International Journal of Organic Chemistry 2012年第3期2卷 295-301页

作者： K. L. Ameta Nitu S. Rathore Biresh Kumar Edith S. Malaga M Manuela Verastegui P Robert H. Gilman B. L. Verma Department of Chemistry FASC Mody Institute of Technology & Science Lakshmangarh India Department of Chemistry MLS University Udaipur India Department of International Health Bloomberg School of Public Health Johns Hopkins University Baltimore USA Department of Microbiology Faculty of Science and Philosophy Universidad Peruana Cayetano Heredia Lima Peru

Substituted 2-benzylidene-1-benzofuran-3-ones are commonly known as aurones. This class of bioactive heterocycles belongs to flavonoid family. The article intends to put forth the rational design and synthesis of a new series of aurones using 3’,5’-dibromo-2’,4’-dihydroxychalcones and copper bromide in presence of DMF-water mixture (8:2, v/v) for the first time. Preliminary bioassay shows that most of compounds have good trypanocidal activity against Trypanosoma cruzi at 10 μg/mL. Few compounds are equally potent to the standard drugs Benznidazole and Nifurtimox. The structures of the newly synthesized products 2a-n were established by elemental analysis, FTIR, 1H NMR, 13C NMR and mass spectroscopic studies.

关键词： Chalcones Aurones Trypanosoma cruzi Chagas Disease Inhibition