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Sample records for screen yields trypanosoma

  1. A target-based high throughput screen yields Trypanosoma brucei hexokinase small molecule inhibitors with antiparasitic activity.

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    Elizabeth R Sharlow

    2010-04-01

    Full Text Available The parasitic protozoan Trypanosoma brucei utilizes glycolysis exclusively for ATP production during infection of the mammalian host. The first step in this metabolic pathway is mediated by hexokinase (TbHK, an enzyme essential to the parasite that transfers the gamma-phospho of ATP to a hexose. Here we describe the identification and confirmation of novel small molecule inhibitors of bacterially expressed TbHK1, one of two TbHKs expressed by T. brucei, using a high throughput screening assay.Exploiting optimized high throughput screening assay procedures, we interrogated 220,233 unique compounds and identified 239 active compounds from which ten small molecules were further characterized. Computation chemical cluster analyses indicated that six compounds were structurally related while the remaining four compounds were classified as unrelated or singletons. All ten compounds were approximately 20-17,000-fold more potent than lonidamine, a previously identified TbHK1 inhibitor. Seven compounds inhibited T. brucei blood stage form parasite growth (0.03

  2. Targeted screening strategies to detect Trypanosoma cruzi infection in children.

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    Michael Z Levy

    2007-12-01

    Full Text Available Millions of people are infected with Trypanosoma cruzi, the causative agent of Chagas disease in Latin America. Anti-trypanosomal drug therapy can cure infected individuals, but treatment efficacy is highest early in infection. Vector control campaigns disrupt transmission of T. cruzi, but without timely diagnosis, children infected prior to vector control often miss the window of opportunity for effective chemotherapy.We performed a serological survey in children 2-18 years old living in a peri-urban community of Arequipa, Peru, and linked the results to entomologic, spatial and census data gathered during a vector control campaign. 23 of 433 (5.3% [95% CI 3.4-7.9] children were confirmed seropositive for T. cruzi infection by two methods. Spatial analysis revealed that households with infected children were very tightly clustered within looser clusters of households with parasite-infected vectors. Bayesian hierarchical mixed models, which controlled for clustering of infection, showed that a child's risk of being seropositive increased by 20% per year of age and 4% per vector captured within the child's house. Receiver operator characteristic (ROC plots of best-fit models suggest that more than 83% of infected children could be identified while testing only 22% of eligible children.We found evidence of spatially-focal vector-borne T. cruzi transmission in peri-urban Arequipa. Ongoing vector control campaigns, in addition to preventing further parasite transmission, facilitate the collection of data essential to identifying children at high risk of T. cruzi infection. Targeted screening strategies could make integration of diagnosis and treatment of children into Chagas disease control programs feasible in lower-resource settings.

  3. Selection and optimization of hits from a high-throughput phenotypic screen against Trypanosoma cruzi.

    Science.gov (United States)

    Keenan, Martine; Alexander, Paul W; Chaplin, Jason H; Abbott, Michael J; Diao, Hugo; Wang, Zhisen; Best, Wayne M; Perez, Catherine J; Cornwall, Scott M J; Keatley, Sarah K; Thompson, R C Andrew; Charman, Susan A; White, Karen L; Ryan, Eileen; Chen, Gong; Ioset, Jean-Robert; von Geldern, Thomas W; Chatelain, Eric

    2013-10-01

    Inhibitors of Trypanosoma cruzi with novel mechanisms of action are urgently required to diversify the current clinical and preclinical pipelines. Increasing the number and diversity of hits available for assessment at the beginning of the discovery process will help to achieve this aim. We report the evaluation of multiple hits generated from a high-throughput screen to identify inhibitors of T. cruzi and from these studies the discovery of two novel series currently in lead optimization. Lead compounds from these series potently and selectively inhibit growth of T. cruzi in vitro and the most advanced compound is orally active in a subchronic mouse model of T. cruzi infection. High-throughput screening of novel compound collections has an important role to play in diversifying the trypanosomatid drug discovery portfolio. A new T. cruzi inhibitor series with good drug-like properties and promising in vivo efficacy has been identified through this process.

  4. Cost-Effectiveness of Blood Donation Screening for Trypanosoma cruzi in Mexico

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    Sánchez-González, Gilberto; Figueroa-Lara, Alejandro; Elizondo-Cano, Miguel; Wilson, Leslie; Novelo-Garza, Barbara; Valiente-Banuet, Leopoldo; Ramsey, Janine M.

    2016-01-01

    An estimated 2 million inhabitants are infected with Chagas disease in Mexico, with highest prevalence coinciding with highest demographic density in the southern half of the country. After vector-borne transmission, Trypanosoma cruzi is principally transmitted to humans via blood transfusion. Despite initiation of serological screening of blood donations or donors for T. cruzi since 1990 in most Latin American countries, Mexico only finally included mandatory serological screening nationwide in official Norms in 2012. Most recent regulatory changes and segmented blood services in Mexico may affect compliance of mandatory screening guidelines. The objective of this study was to calculate the incremental cost-effectiveness ratio for total compliance of current guidelines from both Mexican primary healthcare and regular salaried worker health service institutions: the Secretary of Health and the Mexican Institute for Social Security. We developed a bi-modular model to analyze compliance using a decision tree for the most common screening algorithms for each health institution, and a Markov transition model for the natural history of illness and care. The incremental cost effectiveness ratio based on life-years gained is US$ 383 for the Secretary of Health, while the cost for an additional life-year gained is US$ 463 for the Social Security Institute. The results of the present study suggest that due to incomplete compliance of Mexico’s national legislation during 2013 and 2014, the MoH has failed to confirm 15,162 T. cruzi infections, has not prevented 2,347 avoidable infections, and has lost 333,483 life-years. Although there is a vast difference in T. cruzi prevalence between Bolivia and Mexico, Bolivia established mandatory blood screening for T.cruzi in 1996 and until 2002 detected and discarded 11,489 T. cruzi -infected blood units and prevented 2,879 potential infections with their transfusion blood screening program. In the first two years of Mexico

  5. Cost-Effectiveness of Blood Donation Screening for Trypanosoma cruzi in Mexico.

    Science.gov (United States)

    Sánchez-González, Gilberto; Figueroa-Lara, Alejandro; Elizondo-Cano, Miguel; Wilson, Leslie; Novelo-Garza, Barbara; Valiente-Banuet, Leopoldo; Ramsey, Janine M

    2016-03-01

    An estimated 2 million inhabitants are infected with Chagas disease in Mexico, with highest prevalence coinciding with highest demographic density in the southern half of the country. After vector-borne transmission, Trypanosoma cruzi is principally transmitted to humans via blood transfusion. Despite initiation of serological screening of blood donations or donors for T. cruzi since 1990 in most Latin American countries, Mexico only finally included mandatory serological screening nationwide in official Norms in 2012. Most recent regulatory changes and segmented blood services in Mexico may affect compliance of mandatory screening guidelines. The objective of this study was to calculate the incremental cost-effectiveness ratio for total compliance of current guidelines from both Mexican primary healthcare and regular salaried worker health service institutions: the Secretary of Health and the Mexican Institute for Social Security. We developed a bi-modular model to analyze compliance using a decision tree for the most common screening algorithms for each health institution, and a Markov transition model for the natural history of illness and care. The incremental cost effectiveness ratio based on life-years gained is US$ 383 for the Secretary of Health, while the cost for an additional life-year gained is US$ 463 for the Social Security Institute. The results of the present study suggest that due to incomplete compliance of Mexico's national legislation during 2013 and 2014, the MoH has failed to confirm 15,162 T. cruzi infections, has not prevented 2,347 avoidable infections, and has lost 333,483 life-years. Although there is a vast difference in T. cruzi prevalence between Bolivia and Mexico, Bolivia established mandatory blood screening for T.cruzi in 1996 and until 2002 detected and discarded 11,489 T. cruzi -infected blood units and prevented 2,879 potential infections with their transfusion blood screening program. In the first two years of Mexico's mandated

  6. Cost-Effectiveness of Blood Donation Screening for Trypanosoma cruzi in Mexico.

    Directory of Open Access Journals (Sweden)

    Gilberto Sánchez-González

    2016-03-01

    Full Text Available An estimated 2 million inhabitants are infected with Chagas disease in Mexico, with highest prevalence coinciding with highest demographic density in the southern half of the country. After vector-borne transmission, Trypanosoma cruzi is principally transmitted to humans via blood transfusion. Despite initiation of serological screening of blood donations or donors for T. cruzi since 1990 in most Latin American countries, Mexico only finally included mandatory serological screening nationwide in official Norms in 2012. Most recent regulatory changes and segmented blood services in Mexico may affect compliance of mandatory screening guidelines. The objective of this study was to calculate the incremental cost-effectiveness ratio for total compliance of current guidelines from both Mexican primary healthcare and regular salaried worker health service institutions: the Secretary of Health and the Mexican Institute for Social Security. We developed a bi-modular model to analyze compliance using a decision tree for the most common screening algorithms for each health institution, and a Markov transition model for the natural history of illness and care. The incremental cost effectiveness ratio based on life-years gained is US$ 383 for the Secretary of Health, while the cost for an additional life-year gained is US$ 463 for the Social Security Institute. The results of the present study suggest that due to incomplete compliance of Mexico's national legislation during 2013 and 2014, the MoH has failed to confirm 15,162 T. cruzi infections, has not prevented 2,347 avoidable infections, and has lost 333,483 life-years. Although there is a vast difference in T. cruzi prevalence between Bolivia and Mexico, Bolivia established mandatory blood screening for T.cruzi in 1996 and until 2002 detected and discarded 11,489 T. cruzi -infected blood units and prevented 2,879 potential infections with their transfusion blood screening program. In the first two years

  7. Yield of nat screening in Slovenia

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    Snežna Levičnik Stezinar

    2012-12-01

    Conclusion: The implementation of NAT screening for three viruses has improved the blood safety. In the five-year period, 31 infectious units that were NAT–only positive were eliminated from the stock.

  8. Visual genome-wide RNAi screening to identify human host factors required for Trypanosoma cruzi infection

    CSIR Research Space (South Africa)

    Genovesio, A

    2011-05-01

    Full Text Available The protozoan parasite Trypanosoma cruzi is the etiologic agent of Chagas disease, a neglected tropical infection that affects millions of people in the Americas. Current chemotherapy relies on only two drugs that have limited efficacy...

  9. The early implementation of Trypanosoma cruzi antibody screening of donors and donations within England: preempting a problem.

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    Kitchen, Alan D; Hewitt, Patricia E; Chiodini, Peter L

    2012-09-01

    Trypanosoma cruzi is a parasitic infection endemic in Central and Southern America, but is spreading into nonendemic countries with migration of infected individuals from endemic countries. The parasite is transmitted by transfusion or transplantation and donation screening is performed routinely in endemic countries to prevent transmission. In situations where migrants from endemic countries have settled in nonendemic countries and present as donors (blood or other cellular products), intervention is required to prevent transfusion or transplantation transmission. A screening program for T. cruzi was developed and has been used successfully for over 10 years that includes donor selection and donation screening. Donor selection criteria to identify specific risk of T. cruzi infection were developed together with laboratory screening of donations for T. cruzi antibodies and the subsequent confirmation of screen reactivity. Since the introduction of T. cruzi screening in England in 1998, a total of 38,585 donors and donations have been screened for T. cruzi antibodies, of which 223 were repeat reactive on screening and referred for confirmation: 206 confirmed negative, 14 inconclusive, and three positive. Since the move in 2005 from donor qualification to donation release testing, 15,536 donations were collected and screened, of which 15,499 (99.8%) were T. cruzi antibody negative and released to inventory. An effective program to minimize risk of the transmission of T. cruzi infection via donations has been developed and implemented. Not only does the program minimize risk of transmission, it also minimizes the cumulative, and needless, loss of donors and donations that would ensue if permanent donor deferral alone was adopted. © 2012 American Association of Blood Banks.

  10. Identification of three classes of heteroaromatic compounds with activity against intracellular Trypanosoma cruzi by chemical library screening.

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    Esther Bettiol

    Full Text Available The development of new drugs against Chagas disease is a priority since the currently available medicines have toxic effects, partial efficacy and are targeted against the acute phase of disease. At present, there is no drug to treat the chronic stage. In this study, we have optimized a whole cell-based assay for high throughput screening of compounds that inhibit infection of mammalian cells by Trypanosoma cruzi trypomastigotes. A 2000-compound chemical library was screened using a recombinant T. cruzi (Tulahuen strain expressing beta-galactosidase. Three hits were selected for their high activity against T. cruzi and low toxicity to host cells in vitro: PCH1, NT1 and CX1 (IC(50: 54, 190 and 23 nM, respectively. Each of these three compounds presents a different mechanism of action on intracellular proliferation of T. cruzi amastigotes. CX1 shows strong trypanocidal activity, an essential characteristic for the development of drugs against the chronic stage of Chagas disease where parasites are found intracellular in a quiescent stage. NT1 has a trypanostatic effect, while PCH1 affects parasite division. The three compounds also show high activity against intracellular T. cruzi from the Y strain and against the related kinetoplastid species Leishmania major and L. amazonensis. Characterization of the anti-T. cruzi activity of molecules chemically related to the three library hits allowed the selection of two compounds with IC(50 values of 2 nM (PCH6 and CX2. These values are approximately 100 times lower than those of the medicines used in patients against T. cruzi. These results provide new candidate molecules for the development of treatments against Chagas disease and leishmaniasis.

  11. Identification of three classes of heteroaromatic compounds with activity against intracellular Trypanosoma cruzi by chemical library screening.

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    Bettiol, Esther; Samanovic, Marie; Murkin, Andrew S; Raper, Jayne; Buckner, Frederick; Rodriguez, Ana

    2009-01-01

    The development of new drugs against Chagas disease is a priority since the currently available medicines have toxic effects, partial efficacy and are targeted against the acute phase of disease. At present, there is no drug to treat the chronic stage. In this study, we have optimized a whole cell-based assay for high throughput screening of compounds that inhibit infection of mammalian cells by Trypanosoma cruzi trypomastigotes. A 2000-compound chemical library was screened using a recombinant T. cruzi (Tulahuen strain) expressing beta-galactosidase. Three hits were selected for their high activity against T. cruzi and low toxicity to host cells in vitro: PCH1, NT1 and CX1 (IC(50): 54, 190 and 23 nM, respectively). Each of these three compounds presents a different mechanism of action on intracellular proliferation of T. cruzi amastigotes. CX1 shows strong trypanocidal activity, an essential characteristic for the development of drugs against the chronic stage of Chagas disease where parasites are found intracellular in a quiescent stage. NT1 has a trypanostatic effect, while PCH1 affects parasite division. The three compounds also show high activity against intracellular T. cruzi from the Y strain and against the related kinetoplastid species Leishmania major and L. amazonensis. Characterization of the anti-T. cruzi activity of molecules chemically related to the three library hits allowed the selection of two compounds with IC(50) values of 2 nM (PCH6 and CX2). These values are approximately 100 times lower than those of the medicines used in patients against T. cruzi. These results provide new candidate molecules for the development of treatments against Chagas disease and leishmaniasis.

  12. Novo processo para triagem de medicamentos na infecção experimental pelo Trypanosoma cruzi Proposal of a new process for screening of drugs in experimental infection with Trypanosoma cruzi

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    Rubens Campos

    1991-08-01

    Full Text Available É proposto processo para a triagem da capacidade terapêutica de medicamentos na infecção experimental pelo Trypanosoma cruzi. O método tem base no emprego de triatomíneos parasitados que se alimentam, decorridos períodos diferentes para haver compatibilização com níveis sangüíneos, em camundongos aos quais foi administrado o fármaco sob apreciação; assim, o tubo digestivo do hemíptero participará como estrutura propícia à avaliação. Em observação inicial, ocorreu utilização do benzonidazol, que se mostrou apenas parcialmente ativo, pelo menos de acordo com a maneira de execução do novo procedimento.We propose a screening process for detection of therapeutic activity of drugs against experimental infection with Trypanosoma cruzi. It is based on the use of infected tryatominae that are fed on mice which have received the study drug. Blood meals are made at different time schedule in order to adapt with serum drug levels. The digestive tube of the hemyptera will, thus, work as a suitable structure for examination. In a initial observation, benzonidazole was used, and was shown to be only partially active at least in the conditions of this new procedure.

  13. Yield and Efficiency of Mental Health Screening: A Comparison of Screening Protocols at Intake to Prison.

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    Martin, Michael S; Potter, Beth K; Crocker, Anne G; Wells, George A; Colman, Ian

    2016-01-01

    The value of screening for mental illness has increasingly been questioned in low prevalence settings due to high false positive rates. However, since false positive rates are related to prevalence, screening may be more effective in higher prevalence settings, including correctional institutions. We compared the yield (i.e. newly detected cases) and efficiency (i.e. false positives) of five screening protocols to detect mental illness in prisons against the use of mental health history taking (the prior approach to detecting mental illness). We estimated the accuracy of the six approaches to detect an Axis I disorder among a sample of 467 newly admitted male inmates (83.1% participation rate). Mental health history taking identified only 41.0% (95% CI 32.1, 50.6) of all inmates with mental illness. Screening protocols identified between 61.9 and 85.7% of all cases, but referred between 2 and 3 additional individuals who did not have a mental illness for every additional case detected compared to the mental health history taking approach. In low prevalence settings (i.e. 10% or less) the screening protocols would have had between 4.6 and 16.2 false positives per true positive. While screening may not be practical in low prevalence settings, it may be beneficial in jails and prisons where the prevalence of mental illness is higher. Further consideration of the context in which screening is being implemented, and of the impacts of policies and clinical practices on the benefits and harms of screening is needed to determine the effectiveness of screening in these settings.

  14. Yield and Efficiency of Mental Health Screening: A Comparison of Screening Protocols at Intake to Prison.

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    Michael S Martin

    Full Text Available The value of screening for mental illness has increasingly been questioned in low prevalence settings due to high false positive rates. However, since false positive rates are related to prevalence, screening may be more effective in higher prevalence settings, including correctional institutions. We compared the yield (i.e. newly detected cases and efficiency (i.e. false positives of five screening protocols to detect mental illness in prisons against the use of mental health history taking (the prior approach to detecting mental illness.We estimated the accuracy of the six approaches to detect an Axis I disorder among a sample of 467 newly admitted male inmates (83.1% participation rate. Mental health history taking identified only 41.0% (95% CI 32.1, 50.6 of all inmates with mental illness. Screening protocols identified between 61.9 and 85.7% of all cases, but referred between 2 and 3 additional individuals who did not have a mental illness for every additional case detected compared to the mental health history taking approach. In low prevalence settings (i.e. 10% or less the screening protocols would have had between 4.6 and 16.2 false positives per true positive.While screening may not be practical in low prevalence settings, it may be beneficial in jails and prisons where the prevalence of mental illness is higher. Further consideration of the context in which screening is being implemented, and of the impacts of policies and clinical practices on the benefits and harms of screening is needed to determine the effectiveness of screening in these settings.

  15. Repositioning FDA Drugs as Potential Cruzain Inhibitors from Trypanosoma cruzi: Virtual Screening, In Vitro and In Vivo Studies

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    Isidro Palos

    2017-06-01

    Full Text Available Chagas disease (CD is a neglected disease caused by the parasite Trypanosoma cruzi, which affects underdeveloped countries. The current drugs of choice are nifurtimox and benznidazole, but both have severe adverse effects and less effectivity in chronic infections; therefore, the need to discover new drugs is essential. A computer-guided drug repositioning method was applied to identify potential FDA drugs (approved and withdrawn as cruzain (Cz inhibitors and trypanocidal effects were confirmed by in vitro and in vivo studies. 3180 FDA drugs were virtually screened using a structure-based approach. From a first molecular docking analysis, a set of 33 compounds with the best binding energies were selected. Subsequent consensus affinity binding, ligand amino acid contact clustering analysis, and ranked position were used to choose four known pharmacological compounds to be tested in vitro. Mouse blood samples infected with trypomastigotes from INC-5 and NINOA strains were used to test the trypanocidal effect of four selected compounds. Among these drugs, one fibrate antilipemic (etofyllin clofibrate and three β-lactam antibiotics (piperacillin, cefoperazone, and flucloxacillin showed better trypanocidal effects (LC50 range 15.8–26.1 μg/mL in comparison with benznidazole and nifurtimox (LC50 range 33.1–46.7 μg/mL. A short-term in vivo evaluation of these compounds showed a reduction of parasitemia in infected mice (range 90–60% at 6 h, but this was low compared to benznidazole (50%. This work suggests that four known FDA drugs could be used to design and obtain new trypanocidal agents.

  16. An Anti-proteome Nanobody Library Approach Yields a Specific Immunoassay for Trypanosoma congolense Diagnosis Targeting Glycosomal Aldolase.

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    Steven Odongo

    2016-02-01

    Full Text Available Infectious diseases pose a severe worldwide threat to human and livestock health. While early diagnosis could enable prompt preventive interventions, the majority of diseases are found in rural settings where basic laboratory facilities are scarce. Under such field conditions, point-of-care immunoassays provide an appropriate solution for rapid and reliable diagnosis. The limiting steps in the development of the assay are the identification of a suitable target antigen and the selection of appropriate high affinity capture and detection antibodies. To meet these challenges, we describe the development of a Nanobody (Nb-based antigen detection assay generated from a Nb library directed against the soluble proteome of an infectious agent. In this study, Trypanosoma congolense was chosen as a model system.An alpaca was vaccinated with whole-parasite soluble proteome to generate a Nb library from which the most potent T. congolense specific Nb sandwich immunoassay (Nb474H-Nb474B was selected. First, the Nb474-homologous sandwich ELISA (Nb474-ELISA was shown to detect experimental infections with high Positive Predictive Value (98%, Sensitivity (87% and Specificity (94%. Second, it was demonstrated under experimental conditions that the assay serves as test-of-cure after Berenil treatment. Finally, this assay allowed target antigen identification. The latter was independently purified through immuno-capturing from (i T. congolense soluble proteome, (ii T. congolense secretome preparation and (iii sera of T. congolense infected mice. Subsequent mass spectrometry analysis identified the target as T. congolense glycosomal aldolase.The results show that glycosomal aldolase is a candidate biomarker for active T. congolense infections. In addition, and by proof-of-principle, the data demonstrate that the Nb strategy devised here offers a unique approach to both diagnostic development and target discovery that could be widely applied to other infectious

  17. Yield of opportunistic targeted screening for type 2 diabetes in primary care: the diabscreen study.

    NARCIS (Netherlands)

    Klein Woolthuis, E.P.; Grauw, W.J.C. de; Gerwen, W.H.E.M. van; Hoogen, H.J.M. van den; Lisdonk, E.H. van de; Metsemakers, J.F.M.; Weel, C. van

    2009-01-01

    PURPOSE: In screening for type 2 diabetes, guidelines recommend targeting high-risk individuals. Our objectives were to assess the yield of opportunistic targeted screening for type 2 diabetes in primary care and to assess the diagnostic value of various risk factors. METHODS: In 11 family practices

  18. Choosing algorithms for TB screening: a modelling study to compare yield, predictive value and diagnostic burden.

    Science.gov (United States)

    Van't Hoog, Anna H; Onozaki, Ikushi; Lonnroth, Knut

    2014-10-19

    To inform the choice of an appropriate screening and diagnostic algorithm for tuberculosis (TB) screening initiatives in different epidemiological settings, we compare algorithms composed of currently available methods. Of twelve algorithms composed of screening for symptoms (prolonged cough or any TB symptom) and/or chest radiography abnormalities, and either sputum-smear microscopy (SSM) or Xpert MTB/RIF (XP) as confirmatory test we model algorithm outcomes and summarize the yield, number needed to screen (NNS) and positive predictive value (PPV) for different levels of TB prevalence. Screening for prolonged cough has low yield, 22% if confirmatory testing is by SSM and 32% if XP, and a high NNS, exceeding 1000 if TB prevalence is ≤0.5%. Due to low specificity the PPV of screening for any TB symptom followed by SSM is less than 50%, even if TB prevalence is 2%. CXR screening for TB abnormalities followed by XP has the highest case detection (87%) and lowest NNS, but is resource intensive. CXR as a second screen for symptom screen positives improves efficiency. The ideal algorithm does not exist. The choice will be setting specific, for which this study provides guidance. Generally an algorithm composed of CXR screening followed by confirmatory testing with XP can achieve the lowest NNS and highest PPV, and is the least amenable to setting-specific variation. However resource requirements for tests and equipment may be prohibitive in some settings and a reason to opt for symptom screening and SSM. To better inform disease control programs we need empirical data to confirm the modeled yield, cost-effectiveness studies, transmission models and a better screening test.

  19. Short report: screening for Trypanosoma cruzi in the blood supply by the Red Cross blood bank in Quito, Ecuador.

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    Grijalva, M J; Chiriboga, R; Racines, J R; Escalante, L; Rowland, E C

    1997-12-01

    The status of Chagas' disease in Ecuador is not clear. In response to reports suggesting the possibility of transfusion-associated transmission of Chagas' disease in the blood bank in Quito, the Ecuadorian Red Cross in collaboration with the Instituto Nacional de Higiene, Zona Norte and the Tropical Disease Institute of Ohio University implemented a pilot Chagas' disease screening of the donated blood in the Quito blood bank. The results of the screening showed a low incidence of seropositivity among the donors (0.01% in 1994, 0.04% in 1995, and 0.02% in 1996) to the Quito blood bank and a higher seropositivity in samples donated to smaller blood banks (0.4% in 1994, 0.28% in 1995, and 0.13% in 1996) located in areas considered endemic, as well as from at least two areas previously considered nonendemic for Chagas' disease. This report highlights the need for a comprehensive evaluation of the prevalence and distribution of Chagas' disease in Ecuador.

  20. Feasibility and yield of screening for non-communicable diseases among treated tuberculosis patients in Peru.

    Science.gov (United States)

    Byrne, A L; Marais, B J; Mitnick, C D; Garden, F L; Lecca, L; Contreras, C; Yauri, Y; Garcia, F; Marks, G B

    2018-01-01

    The increasing prevalence of non-communicable diseases (NCDs) poses a major challenge to low- and middle-income countries. Patients' engagement with health services for anti-tuberculosis treatment provides an opportunity for screening for NCDs and for linkage to care. We explored the feasibility and yield of screening for NCDs in patients treated for tuberculosis (TB) in Lima, Peru, as part of a study focused on chronic respiratory sequelae. A representative sample of community controls was recruited from the same geographical area. Screening entailed taking a medical history and performing ambulatory blood pressure measurement and urinalysis. A total of 177 participants with previous TB (33 with multidrug-resistant TB) and 161 community controls were evaluated. There was an almost four-fold increased prevalence of self-reported diabetes mellitus (DM) in the TB group (adjusted prevalence ratio 3.66, 95%CI 1.68-8.01). Among those without self-reported DM, 3.3% had glycosuria, with a number needed to screen (NNS) of 31. The NNS to find one (new) case of hypertension or proteinuria in the TB group was respectively 24 and 5. Patient-centred care that includes pragmatic NCD screening is feasible in TB patients, and the treatment period provides a good opportunity to link patients to ongoing care.

  1. Low Yield of Chest Radiography in a Large Tuberculosis Screening Program1

    Science.gov (United States)

    Pollock, Nira R.

    2010-01-01

    Purpose: To assess the frequency and spectrum of abnormalities on routine screening chest radiographs in the pre-employment evaluation of health care workers with positive tuberculin skin test (TST) results. Materials and Methods: The institutional review board approved this HIPAA-compliant retrospective study and waived the need for written informed patient consent. Chest radiographic reports of all 2586 asymptomatic individuals with positive TST results who underwent pre-employment evaluation between January 1, 2003, and December 31, 2007, were evaluated to determine the frequency of detection of evidence of active tuberculosis (TB) or latent TB infection (LTBI) and the spectrum of imaging findings. All chest radiographs interpreted as positive were reviewed by an experienced board-certified radiologist. If there was a discrepancy between the two readings, a second experienced radiologist served as an independent and final arbiter. Any follow-up chest radiographs or computed tomographic images that had been acquired by employee health services or by the employee’s private physician as a result of a suspected abnormality detected at initial screening were also evaluated. Results: Of the 159 (6.1%) chest radiographic examinations that yielded abnormal results, there were no findings that were consistent with active TB. There were 92 cases of calcified granulomas, calcified lymph nodes, or both; 25 cases of apical pleural thickening; 16 cases of fibrous scarring; and 31 cases of noncalcified nodules. All cases of fibrous scarring involved an area smaller than 2 cm2. All noncalcified nodules were 4 mm in diameter or smaller, with the exception of one primary lung malignancy and one necrotizing granuloma (negative for acid-fast bacilli) that grew Mycobacterium kansasii on culture. Conclusion: Universal chest radiography in a large pre-employment TB screening program was of low yield in the detection of active TB or increased LTBI reactivation risk, and it provided

  2. Performance of six diagnostic tests to screen for Chagas disease in blood banks andprevalence of Trypanosoma cruzi infection among donors with inconclusive serologyscreening based on the analysis of epidemiological variables.

    Science.gov (United States)

    Pereira, Gilberto de Araujo; Louzada-Neto, Francisco; Barbosa, Valdirene de Fátima; Ferreira-Silva, Márcia Maria; de Moraes-Souza, Helio

    2012-01-01

    The frequent occurrence of inconclusive serology in blood banks and the absence of a gold standard test for Chagas'disease led us to examine the efficacy of the blood culture test and five commercial tests (ELISA, IIF, HAI, c-ELISA, rec-ELISA) used in screening blood donors for Chagas disease, as well as to investigate the prevalence of Trypanosoma cruzi infection among donors with inconclusive serology screening in respect to some epidemiological variables. To obtain estimates of interest we considered a Bayesian latent class model with inclusion of covariates from the logit link. A better performance was observed with some categories of epidemiological variables. In addition, all pairs of tests (excluding the blood culture test) presented as good alternatives for both screening (sensitivity > 99.96% in parallel testing) and for confirmation (specificity > 99.93% in serial testing) of Chagas disease. The prevalence of 13.30% observed in the stratum of donors with inconclusive serology, means that probably most of these are non-reactive serology. In addition, depending on the level of specific epidemiological variables, the absence of infection can be predicted with a probability of 100% in this group from the pairs of tests using parallel testing. The epidemiological variables can lead to improved test results and thus assist in the clarification of inconclusive serology screening results. Moreover, all combinations of pairs using the five commercial tests are good alternatives to confirm results.

  3. Screening of high-yield GTF yeast by N+-implantation

    International Nuclear Information System (INIS)

    Gao Yanhong; Lv Jiaping; Liu Lu; Li Shurong

    2009-01-01

    In this study, one of the highest chromium-resistant strain was screened from 12 tested brewer's yeast. N + ion implantation was used to mutate this yeast and screened high-yield GTF yeast strains with Chromium tolerance method. The mutagenesis was conducted by 50 KeV N + ion implantation with the doses of 1 x 2.6 x 10 13 , 2 x 2.6 x 10 13 , 3 x 2.6 x 10 13 , 4 x 2.6 x 10 13 , 5 x 2.6 x 10 13 and 6 x 2.6 x 10 13 ion/cm 2 . Results showed that the optimum dose was 4 x 2.6 x 10 13 ion/cm 2 , and a strain M11-1A11 of high-producing GTF was obtained. Its organic Cr content was increased by 22.4% than the original strain. Its fermentation property was stable after 5 generation transfer inoculation. (authors)

  4. Screening of post emergence herbicides for weed control in cotton (GOSSYPIUM HIRSUTUM) and their effect on yield and yield components

    International Nuclear Information System (INIS)

    Hussain, N.; Khan, M.B.; Khan, M.A.; Hameed, R.A.

    2005-01-01

    Response of varying herbicides at different levels: round up 490 G/L at the rate of 4.7 L ha/sup -1/ and 1.5 L ha/sup -1/ (Glyphosat) and Gramaxone 20 EC (Paraquat) at the rate of 2.5 L ha/sup -1/ against untreated (control, were investigated to cotton cultivar CIM-473 under field conditions during Kharif 2002 at Agronomic Research Area. Central Cotton Research Institute, Multan. Significant control of weeds and increase in yield and yield contributing factors were observed. It was indicated that maximum yield and weed control were obtained by using Round up (Glyphosate) at the rate of 4.7 L ha/sup -1/ as compared to other treatments including untreated (control). Average boll weight was not significant among treatments but significant against control. Maximum net profit was obtained from Round up 490 G/L when treated at the rate of 4.7 L ha/sup -1/ than all other treatments. (author)

  5. Computer tomography colonography participation and yield in patients under surveillance for 6-9 mm polyps in a population-based screening trial

    NARCIS (Netherlands)

    Tutein Nolthenius, Charlotte J.; Boellaard, Thierry N.; de Haan, Margriet C.; Nio, C. Yung; Thomeer, Maarten G. J.; Bipat, Shandra; Montauban van Swijndregt, Alexander D.; van de Vijver, Marc J.; Biermann, Katharina; Kuipers, Ernst J.; Dekker, Evelien; Stoker, Jaap

    2016-01-01

    Surveillance CT colonography (CTC) is a viable option for 6-9 mm polyps at CTC screening for colorectal cancer. We established participation and diagnostic yield of surveillance and determined overall yield of CTC screening. In an invitational CTC screening trial 82 of 982 participants harboured 6-9

  6. Screening with nuclear techniques for yield and N2 fixation in mung bean in Thailand

    International Nuclear Information System (INIS)

    Boonkerd, N.; Wadisrisuk, P.; Siripin, S.; Murakami, T.; Danso, S.K.A.

    1998-01-01

    For a farmer to reap benefit from mung bean's (Vigna radiata) capacity to fix N 2 , the crop's requirement for N must come mainly from the atmosphere through symbiotic fixation in the root nodules. The aim of this study was to evaluate recommended mung-bean cultivars and advanced breeding lines, and identify high fixers. Preliminary investigations with the 15 N natural-abundance method indicated its utility for measuring N 2 fixation, and the examination of five recommended cultivars and two advanced breeding lines of mung using the 15 N-dilution method showed diversity in N 2 fixation and yield. More than 400 lines of mung bean were screened in soil in cement containers for growth, nodulation, N accumulation and N 2 fixation at 35 days after planting, with the natural-abundance method used to determine N 2 fixation. Genetic variability was observed for all characteristics. Estimates of fixed N ranged from 0-300 mg N/plant. Whereas some lines obtained N mainly from fixation, recommended cultivars apparently obtained their N mainly from soil. The data are discussed in terms of reliability of the 15 N natural-abundance method

  7. Household Contact Screening and Yield of Tuberculosis Cases-A Clinic Based Study in Chennai, South India.

    Directory of Open Access Journals (Sweden)

    Dina Nair

    Full Text Available Contact investigation is an active case finding strategy to increase detection of Tuberculosis (TB and a key component of TB control programs. The household contacts are at a higher risk of exposure than members of the general population. The information on the value and yield of household contact screening and the approaches used in high incidence settings like India is limited.To evaluate the yield of active case finding in household contacts of newly diagnosed smear positive TB patients and the factors associated with increased yield.Retrospective record review of the household contacts of newly diagnosed sputum smear positive patients (index case enrolled in a clinical trial at National Institute of Research in Tuberculosis, Chennai during the period 2007-2014. A sequential screening algorithm with chest x-ray followed by symptom screen was employed to identify presumptive TB patients.643 household contacts of 280 index TB patients were identified out of which 544 (85% consented for screening. 71/544 (13% patients had an abnormal chest radiograph and out of them 70% were symptomatic. A total of 29/544 (5.3% contacts were found to have TB among whom 23/29 (79% were sputum smear positive. The number needed to screen (NNS to identify a new TB case among all household contacts was 19 and among those with an abnormal CXR was 02. Age group > 44 years, male gender and siblings of the index case was associated with abnormal chest radiograph whereas age group between 15-44 was significantly associated with developing TB disease among household contacts.Active screening among household contacts is an effective way to improve TB case detection. The yield for new TB cases among contacts with abnormal x-ray was high in this study and the use of Chest X-rays in combination with symptom screen is recommended.

  8. Neonatal screening for inborn errors of metabolism: cost, yield and outcome.

    Science.gov (United States)

    Pollitt, R J; Green, A; McCabe, C J; Booth, A; Cooper, N J; Leonard, J V; Nicholl, J; Nicholson, P; Tunaley, J R; Virdi, N K

    1997-01-01

    OBJECTIVES. To systematically review the literature on inborn errors of metabolism, neonatal screening technology and screening programmes in order to analyse the costs and benefits of introducing screening based on tandem mass-spectrometry (tandem MS) for a wide range of disorders of amino acid and organic acid metabolism in the UK. To evaluate screening for cystic fibrosis, Duchenne muscular dystrophy and other disorders which are tested on an individual basis. HOW THE RESEARCH WAS CONDUCTED. Systematic searches were carried out of the literature on inborn errors of metabolism, neonatal screening programmes, tandem MS-based neonatal screening technology, economic evaluations of neonatal screening programmes and psychological aspects of neonatal screening. Background material on the biology of inherited metabolic disease, the basic philosophy, and the history and current status of the UK screening programme was also collected. Relevant papers in the grey literature and recent publications were identified by hand-searching. Each paper was graded. For each disease an aggregate grade for the state of knowledge in six key areas was awarded. Additional data were prospectively collected on activity and costs in UK neonatal screening laboratories, and expert clinical opinion on current treatment modalities and outcomes. These data were used to construct a decision-analysis model of neonatal screening technologies, comparing tandem MS with the existing phenylketonuria screening methods. This model determined the cost per additional case identified and, for each disease, the additional treatment costs per case, and the cost per life-year saved. All costs and benefits were discounted at 6% per annum. One-way sensitivity analysis was performed showing the effect of varying the discount rate, the incidence rate of each disorder, the number of neonates screened and the cost of tandem MS, on the cost per life-year gained. RESEARCH FINDINGS. The UK screening programmes for

  9. Taxonomy Icon Data: Trypanosoma brucei [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available Trypanosoma brucei Trypanosoma brucei Trypanosoma_brucei_L.png Trypanosoma_brucei_NL.png Trypanoso...ma_brucei_S.png Trypanosoma_brucei_NS.png http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Trypanoso...ma+brucei&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Trypanosoma+brucei&t=NL http://bioscie...ncedbc.jp/taxonomy_icon/icon.cgi?i=Trypanosoma+brucei&t=S http://biosciencedbc.jp.../taxonomy_icon/icon.cgi?i=Trypanosoma+brucei&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=121 ...

  10. Trypanosoma cruzi strain TcIV infects raccoons from Illinois

    Directory of Open Access Journals (Sweden)

    Cailey Vandermark

    Full Text Available BACKGROUND The northern limits of Trypanosoma cruzi across the territory of the United States remain unknown. The known vectors Triatoma sanguisuga and T. lecticularia find their northernmost limits in Illinois; yet, earlier screenings of those insects did not reveal the presence of the pathogen, which has not been reported in vectors or reservoir hosts in this state. OBJECTIVES Five species of medium-sized mammals were screened for the presence of T. cruzi. METHODS Genomic DNA was isolated from heart, spleen and skeletal muscle of bobcats (Lynx rufus, n = 60, raccoons (Procyon lotor, n = 37, nine-banded armadillos (Dasypus novemcinctus, n = 5, Virginia opossums (Didelphis virginiana, n = 3, and a red fox (Vulpes vulpes. Infections were detected targeting DNA from the kinetoplast DNA minicircle (kDNA and satellite DNA (satDNA. The discrete typing unit (DTU was determined by amplifying two gene regions: the Spliced Leader Intergenic Region (SL, via a multiplex polymerase chain reaction, and the 24Sα ribosomal DNA via a heminested reaction. Resulting sequences were used to calculate their genetic distance against reference DTUs. FINDINGS 18.9% of raccoons were positive for strain TcIV; the rest of mammals tested negative. MAIN CONCLUSIONS These results confirm for the first time the presence of T. cruzi in wildlife from Illinois, suggesting that a sylvatic life cycle is likely to occur in the region. The analyses of sequences of SL suggest that amplicons resulting from a commonly used multiplex reaction may yield non-homologous fragments.

  11. Use, appropriateness, and diagnostic yield of screening colonoscopy: an international observational study (EPAGE)

    DEFF Research Database (Denmark)

    Burnand, B; JK, Harris; Wietlisbach, V

    2006-01-01

    was asked of all centers, it is possible that not all consecutive patients were included. Participating centers were a convenience sample and thus may not be representative. CONCLUSIONS: About 1 of 10 colonoscopies were performed for screening, preferentially in middle-aged individuals. A higher diagnostic...

  12. Screening on the high yield validamycin producing strain by implantation with N+ and Ti+ ion source

    International Nuclear Information System (INIS)

    Yu Long; An Xiao

    2007-01-01

    In order to compared the mutagenic effects of the validamycin producing the strain (Streptomyces hygroscopicus var. Jingganggensis Yen.) was implanted with two kinds of ion sources. The results showed that when two kinds of ion sources implanted into the strain by turns, more positive mutants and higher yield would be acquired. Using this method, a high-yielding strain B1-3 was obtained, which produce the titer of validamycin A of 21514, and was 54.4% higher than that of the original strain. (authors)

  13. Screening for ketosis using multiple logistic regression based on milk yield and composition.

    Science.gov (United States)

    Kayano, Mitsunori; Kataoka, Tomoko

    2015-11-01

    Multiple logistic regression was applied to milk yield and composition data for 632 records of healthy cows and 61 records of ketotic cows in Hokkaido, Japan. The purpose was to diagnose ketosis based on milk yield and composition, simultaneously. The cows were divided into two groups: (1) multiparous, including 314 healthy cows and 45 ketotic cows and (2) primiparous, including 318 healthy cows and 16 ketotic cows, since nutritional status, milk yield and composition are affected by parity. Multiple logistic regression was applied to these groups separately. For multiparous cows, milk yield (kg/day/cow) and protein-to-fat (P/F) ratio in milk were significant factors (Pketosis. For primiparous cows, lactose content (%), solid not fat (SNF) content (%) and milk urea nitrogen (MUN) content (mg/dl) were significantly associated with ketosis (Pketosis, provided the sensitivity, specificity and AUC values of (1) 0.711, 0.726 and 0.781; and (2) 0.678, 0.767 and 0.738, respectively.

  14. Use of 15N dilution method for screening soybean lines with high yield and high nitrogen fixation ability

    International Nuclear Information System (INIS)

    Li Haixian; Li Xinmin; Danso, S.K.A.

    1998-01-01

    15 N dilution method was used for screening soybean lines with high nitrogen fixation ability. Screened lines 1005, 8502, 2096, 943, 1454 and Dongnong-42 have high nitrogen fixation ability with their % Ndfa of about 70%. 1454 and 1555 are both high yield and high nitrogen fixation lines. The ability of nitrogen fixation was not related to the yield, but related to maturing time. The cultivars with different maturing time have different levels of nitrogen fixation ability. The longer the maturing period is, the greater the ability of nitrogen fixation it has. There were ten cultivars or lines used in the test of 1992 and 1994. Although the weather condition were greatly different between the two years the results of seven cultivars or lines were the same, indicating that nitrogen fixation ability of the soybean is stable with years. Using 15 N dilution method to estimate nitrogen fixation ability of soybean is reliable, however, the % Ndfa of lines 8502 and 2096 increased by 19% in 1994, a rainy year, indicating that a change in % Ndfa with a few varieties maybe caused by weather

  15. Yield of facility-based verbal screening amongst household contacts of patients with multi-drug resistant tuberculosis in Pakistan

    Directory of Open Access Journals (Sweden)

    Ejaz Qadeer

    2017-05-01

    Full Text Available Background: Household contacts of multidrug-resistant tuberculosis (MDR-TB patients are at a high risk of getting infected with TB/MDR-TB, therefore symptomatic or vulnerable individuals should be screened and treated early. Methods: A cross-sectional study was conducted among household contacts of MDR-TB patients in three high-burden TB sites in Pakistan from July 2013 to June 2014. MDR-TB index patients were asked to provide a list of all members of their household and were asked whether any of them had TB symptoms such as productive cough, fever, weight loss and night sweat (“facility-based verbal screening”. Symptomatic contacts were defined as presumptive TB cases and were invited for investigations at the facility. Those who did not come were paid a home-visit. Confirmed TB/MDR-TB patients were registered in the nearest treatment facility. Results: Of 209 MDR-TB index patients, 1467 household contacts were identified and screened, 95 of them children < 5 years. Of these 172 (12% were symptomatic. Most common symptoms were cough 157 (91% and fever 107 (62%. 58 (34% presumptive TB contacts were not investigated. Of total contacts, 56 (3.8% were diagnosed with TB, among them 54(96% with MDR-TB and 2(4% with drug-susceptible-TB. The number needed to screen (NNS to identify a new MDR-TB case among adult household contacts was 27 and among presumptive adult and pediatric TB contacts was three. All 56 confirmed patients were registered for treatment. Conclusion: Screening household contacts of MDR-TB index cases may be considered a feasible and high yield option, in high-burden, low-resource settings within Pakistan. The number of presumptive TB contacts required to screen to identify a new MDR-TB case was unusually low, indicating an effective strategy that could easily be scaled-up. The screening and management of vulnerable adults and children living with patients having TB of any form is a major priority in the combined efforts

  16. Screening for eri silkworm (Samia ricini Donovan ecoraces using morphological characters, growth, yields, and ISSR marker

    Directory of Open Access Journals (Sweden)

    Duanpen Wongsorn

    2015-10-01

    Full Text Available The selection of eri silkworm ecoraces with high yield and distinct morphological characters is necessary for variety improvement. The five ecoraces SaKKU1, SaKKU2, SaKKU3, SaKKU4 and SaKKU5 were derived mostly by international academic cooperation. They were cultured using castor leaves of TCO 101 cultivar as food plant at 25±2°C, 80±5% R.H. Based on morphological characters, they are similar, except the body of the 5th instar larva of SaKKU1 is clearly covered with more creamy white powder and the mature larva has a shiny dominant yellow color. The duration of the life cycle among ecoraces was also similar; 46-53 days (SaKKU1, 42-53 days (SaKKU2, 42-52 days (SaKKU3, 40-56 days (SaKKU4 and 41-52 days (SaKKU5. SaKKU1 had the highest survival rate at larval stage (1st – 5th instar (100.00% and larva (1st – 5th instar - adult (88.89%, including the predominant heaviest average larva weight of all instars, 0.0317 g (2nd instar, 0.2206 g (3rd instar, 1.0788 g (4th instar, 4.0102 g (5th instar, and 8.9940 g (5 days of 5th instar, which was significantly different (P<0.05 to other ecoraces. Moreover, this ecorace gave the highest average yields: fresh cocoon weight (3.8016 g, pupa weight (3.2532 g, shell weight (0.5287 g, shell ratio (14.01%, fresh cocoon weight/10,000 larvae (38.01 kg, eggs/moth (531.13 eggs, total eggs (6,375.27 eggs and total hatching eggs (6,006.13 eggs, which was also significantly different (P<0.05 than other ecoraces. Of those properties, especially survival rates and yields, this ecorace (SaKKU1 is favored for further varietal improvement program. In parallel, genetic relationship analysis of eri silkworm ecoraces using inter-simple sequence repeat (ISSR technique was also carried out. The result revealed from dendrogram analysis that SaKKU1 was the farthest distance than other ecoraces, especially against SaKKU3. Based on all above results, the SaKKU1 ecorace was considered to be the most suitable for heat tolerant

  17. Screening of synthetic phage display scFv libraries yields competitive ligands of human leptin receptor.

    Science.gov (United States)

    Molek, Peter; Vodnik, Miha; Strukelj, Borut; Bratkovič, Tomaž

    2014-09-26

    Initially considered the main endogenous anorexigenic factor, fat-derived leptin turned out to be a markedly pleiotropic hormone, influencing diverse physiological processes. Moreover, hyperleptinemia in obese individuals has been linked to the onset or progression of serious disorders, such as cancer, autoimmune diseases, and atherosclerosis, and antagonizing peripheral leptin's signalization has been shown to improve these conditions. To develop an antibody-based leptin antagonist we have devised a tailored panning procedure and screened two phage display libraries of single chain variable antibody fragments (scFvs) against recombinant leptin receptor. One of the scFvs was expressed in Escherichia coli and its interaction with leptin receptor was characterized in more detail. It was found to recognize a discontinuous epitope and to compete with leptin for receptor binding with IC50 and Kd values in the nanomolar range. The reported scFv represents a lead for development of leptin antagonists that may ultimately find use in therapy of various hyperleptinemia-related disorders. Copyright © 2014 Elsevier Inc. All rights reserved.

  18. Yield of coeliac screening in abdominal pain-associated functional gastrointestinal system disorders.

    Science.gov (United States)

    Kansu, Aydan; Kuloğlu, Zarife; Demir, Arzu; Yaman, Aytaç

    2015-11-01

    Chronic abdominal pain (CAP) in childhood is common and in the majority functional. While CAP is one of the complaints of coeliac disease (CD), whether CAP as a sole complaint is indicative of CD is unclear. Our aim was to evaluate the relationship between CAP and CD. The study was conducted on 1047 children (61.1% female, mean age 9.6 ± 4.1 years) with CAP. Patients were evaluated according to the Rome III criteria. Patients with alarm symptoms and conditions known to be associated with CD were excluded. Patients were screened for CD using a rapid tissue transglutaminase (tTG) test; positive cases were tested by tTG ELISA, and duodenal biopsies were obtained if tTG was above the normal limit. Functional dyspepsia (FD), irritable bowel syndrome (IBS) and functional abdominal pain (FAP) were diagnosed in 384 (36.7%), 274 (26.2%) and 389 (37.2%) patients, respectively. In 13 patients, the tTG rapid test was positive; 10 were also positive for tTG by ELISA and histopathological evaluations diagnosed CD in all 10 patients. The overall prevalence of CD was 0.95% (2.2%, 0.5% and 0.5% in patients with IBS, FD and FAP, respectively). The prevalence of CD in patients with IBS was higher than expected but with borderline statistical significance (P = 0.053). CD is found as common in children with FD and FAP as in the general population. CD was more commonly diagnosed in IBS patients with borderline statistical significance. We suggest that particular attention be paid to children with IBS. © 2015 The Authors. Journal of Paediatrics and Child Health © 2015 Paediatrics and Child Health Division (Royal Australasian College of Physicians).

  19. The comparative study on screening of pleurotus polysaccharide high-yield strains by use of ion beam implantation and composite mutagenesis

    International Nuclear Information System (INIS)

    Wang Lianfeng; Chen Henglei; Zhang Jun; Zeng Xianxian

    2009-01-01

    In order to screen pleurotus mycelium polysaccharide high-yield strains, the comparative study was made by use of ion beam implantation and composite mutagenesis before screening. The treating mycelium pellet of pleurotus ferulae tentatively with ion beam implantation was performed at the first. Two polysaccharide high-yield strains, PFPH-1and PFPH-2, were selected using fermentation quantitative screening after auxotrophy qualitative primary screening. It has been found that the polysaccharide yield of the mutants is 551.80mg/L and 659.46mg/L respectively,which increases by 46.55% and 75.14% respectively compared to that of initial strain. Then PFPH-1and PFPH-2, as the original strain, is exposed to ultraviolet light and is suffered by additive of LiCl respectively. The results indicate that the polysaccharide yield of strains 1,9 and 10 decreases by 27%, 38% and 37% respectively compared to that of PFPH-1 meanwhile the polysaccharide yield of strain 17 decreases by 28% compared to that of PFPH-2 after high-flux qualitative primary screening. In this study, composite mutagenesis with exposure of ultra-violet and additive of lithium chloride shows some negative effects. (authors)

  20. Phylogenetic position of the giant anuran trypanosomes Trypanosoma chattoni, Trypanosoma fallisi, Trypanosoma mega, Trypanosoma neveulemairei, and Trypanosoma ranarum inferred from 18S rRNA gene sequences.

    Science.gov (United States)

    Martin, Donald S; Wright, André-Denis G; Barta, John R; Desser, Sherwin S

    2002-06-01

    Phylogenetic relationships within the kinetoplastid flagellates were inferred from comparisons of small-subunit ribosomal RNA gene sequences. These included 5 new gene sequences, Trypanosoma fallisi (2,239 bp), Trypanosoma chattoni (2,180 bp), Trypanosoma mega (2,211 bp), Trypanosoma neveulemairei (2,197 bp), and Trypanosoma ranarum (2,203 bp). Trees produced using maximum-parsimony and distance-matrix methods (least-squares, neighbor-joining, and maximum-likelihood), supported by strong bootstrap and quartet-puzzle analyses, indicated that the trypanosomes are a monophyletic group that divides into 2 major lineages, the salivarian trypanosomes and the nonsalivarian trypanosomes. The nonsalivarian trypanosomes further divide into 2 lineages, 1 containing trypanosomes of birds, mammals, and reptiles and the other containing trypanosomes of fish, reptiles, and anurans. Among the giant trypanosomes, T. chattoni is clearly shown to be distantly related to all the other anuran trypanosome species. Trypanosoma mega is closely associated with T. fallisi and T. ranarum, whereas T. neveulemairei and Trypanosoma rotatorium are sister taxa. The branching order of the anuran trypanosomes suggests that some toad trypanosomes may have evolved by host switching from frogs to toads.

  1. Yield of chest X-ray tuberculosis screening of immigrants during the European refugee crisis of 2015: a single-centre experience.

    Science.gov (United States)

    Weinrich, Julius Matthias; Diel, Roland; Sauer, Markus; Henes, Frank Oliver; Meywald-Walter, Karen; Adam, Gerhard; Schön, Gerhard; Bannas, Peter

    2017-08-01

    Our aim was to determine the prevalence of tuberculosis (TB), the number needed to screen (NNS), and the diagnostic accuracy of chest X-ray (CXR) screening to detect active pulmonary TB during the 2015 European refugee crisis. We evaluated data of all refugees who underwent CXR screening in a single-centre of one German metropolitan area in 2015. We determined the prevalence of TB, NNS, and accuracy of CXR to detect active pulmonary TB. Reference method for active TB was the database of all definite TB cases registered at the Department of Public Health. A total of 17,487 immigrants underwent single-centre CXR screening in 2015; prevalence of definite pulmonary TB was 0.103%. The NNS for detecting one case of active pulmonary TB was 1749. CXR had a sensitivity of 55.6% [95% confidence interval (CI) 30.8-78.5%) and a specificity 98.3% (CI 98.1-98.5%) to reveal one case of active TB. Our single-centre study indicates that chest X-ray screening for TB during the 2015 European refugee crisis was of low yield due the low prevalence of TB and high number needed to screen, thus implicating the need for improved screening algorithms adapted to the overwhelming number of refugees. • Prevalence of pulmonary tuberculosis (TB) among refugees in 2015 was low (0.103%). • The number needed to screen to detect one case of active pulmonary TB was 1749. • Tuberculosis X-ray screening resulted in a low sensitivity and high specificity. • Tuberculosis X-ray screening during the European refugee crisis is of low yield. • Improved screening algorithms are needed due to the overwhelming the number of refugees.

  2. Screens

    OpenAIRE

    2016-01-01

    This Sixth volume in the series The Key Debates. Mutations and Appropriations in European Film Studies investigates the question of screens in the context both of the dematerialization due to digitalization and the multiplication of media screens. Scholars offer various infomations and theories of topics such as the archeology of screen, film and media theories, contemporary art, pragmatics of new ways of screening (from home video to street screening).

  3. Comparing yield and relative costs of WHO TB screening algorithms in selected risk groups among people aged 65 years and over in China, 2013.

    Directory of Open Access Journals (Sweden)

    Canyou Zhang

    Full Text Available To calculate the yield and cost per diagnosed tuberculosis (TB case for three World Health Organization screening algorithms and one using the Chinese National TB program (NTP TB suspect definitions, using data from a TB prevalence survey of people aged 65 years and over in China, 2013.This was an analytic study using data from the above survey. Risk groups were defined and the prevalence of new TB cases in each group calculated. Costs of each screening component were used to give indicative costs per case detected. Yield, number needed to screen (NNS and cost per case were used to assess the algorithms.The prevalence survey identified 172 new TB cases in 34,250 participants. Prevalence varied greatly in different groups, from 131/100,000 to 4651/ 100,000. Two groups were chosen to compare the algorithms. The medium-risk group (living in a rural area: men, or previous TB case, or close contact or a BMI <18.5, or tobacco user had appreciably higher cost per case (USD 221, 298 and 963 in the three algorithms than the high-risk group (all previous TB cases, all close contacts. (USD 72, 108 and 309 but detected two to four times more TB cases in the population. Using a Chest x-ray as the initial screening tool in the medium risk group cost the most (USD 963, and detected 67% of all the new cases. Using the NTP definition of TB suspects made little difference.To "End TB", many more TB cases have to be identified. Screening only the highest risk groups identified under 14% of the undetected cases,. To "End TB", medium risk groups will need to be screened. Using a CXR for initial screening results in a much higher yield, at what should be an acceptable cost.

  4. Yield of chest X-ray tuberculosis screening of immigrants during the European refugee crisis of 2015: a single-centre experience

    Energy Technology Data Exchange (ETDEWEB)

    Weinrich, Julius Matthias; Sauer, Markus; Henes, Frank Oliver; Adam, Gerhard; Bannas, Peter [University Medical Center Hamburg-Eppendorf, Department of Diagnostic and Interventional Radiology and Nuclear Medicine, Hamburg (Germany); Diel, Roland [University Medical Hospital Schleswig-Holstein, Airway Research Center North (ARCN), Institute for Epidemiology, Kiel (Germany); Meywald-Walter, Karen [Public Health Department Hamburg Central, Hamburg (Germany); Schoen, Gerhard [University Medical Center Hamburg-Eppendorf, Department of Medical Biometry and Epidemiology, Hamburg (Germany)

    2017-08-15

    Our aim was to determine the prevalence of tuberculosis (TB), the number needed to screen (NNS), and the diagnostic accuracy of chest X-ray (CXR) screening to detect active pulmonary TB during the 2015 European refugee crisis. We evaluated data of all refugees who underwent CXR screening in a single-centre of one German metropolitan area in 2015. We determined the prevalence of TB, NNS, and accuracy of CXR to detect active pulmonary TB. Reference method for active TB was the database of all definite TB cases registered at the Department of Public Health. A total of 17,487 immigrants underwent single-centre CXR screening in 2015; prevalence of definite pulmonary TB was 0.103%. The NNS for detecting one case of active pulmonary TB was 1749. CXR had a sensitivity of 55.6% 95% confidence interval (CI 30.8-78.5%) and a specificity 98.3% (CI 98.1-98.5%) to reveal one case of active TB. Our single-centre study indicates that chest X-ray screening for TB during the 2015 European refugee crisis was of low yield due the low prevalence of TB and high number needed to screen, thus implicating the need for improved screening algorithms adapted to the overwhelming number of refugees. (orig.)

  5. Yield of chest X-ray tuberculosis screening of immigrants during the European refugee crisis of 2015: a single-centre experience

    International Nuclear Information System (INIS)

    Weinrich, Julius Matthias; Sauer, Markus; Henes, Frank Oliver; Adam, Gerhard; Bannas, Peter; Diel, Roland; Meywald-Walter, Karen; Schoen, Gerhard

    2017-01-01

    Our aim was to determine the prevalence of tuberculosis (TB), the number needed to screen (NNS), and the diagnostic accuracy of chest X-ray (CXR) screening to detect active pulmonary TB during the 2015 European refugee crisis. We evaluated data of all refugees who underwent CXR screening in a single-centre of one German metropolitan area in 2015. We determined the prevalence of TB, NNS, and accuracy of CXR to detect active pulmonary TB. Reference method for active TB was the database of all definite TB cases registered at the Department of Public Health. A total of 17,487 immigrants underwent single-centre CXR screening in 2015; prevalence of definite pulmonary TB was 0.103%. The NNS for detecting one case of active pulmonary TB was 1749. CXR had a sensitivity of 55.6% 95% confidence interval (CI 30.8-78.5%) and a specificity 98.3% (CI 98.1-98.5%) to reveal one case of active TB. Our single-centre study indicates that chest X-ray screening for TB during the 2015 European refugee crisis was of low yield due the low prevalence of TB and high number needed to screen, thus implicating the need for improved screening algorithms adapted to the overwhelming number of refugees. (orig.)

  6. Diagnostic yield, interpretation, and clinical utility of mutation screening of sarcomere encoding genes in Danish hypertrophic cardiomyopathy patients and relatives

    DEFF Research Database (Denmark)

    Andersen, Paal Skytt; Havndrup, Ole; Hougs, Lotte

    2008-01-01

    persons. Index patients were screened for mutations in all coding regions of 10 sarcomere genes (MYH7, MYL3, MYBPC3, TNNI3, TNNT2, TPM1, ACTC, CSRP3, TCAP, and TNNC1) and five exons of TTN. Relatives were screened for presence of minor or major diagnostic criteria for HCM and tracking of DNA variants...

  7. Mutation and screening of high-alcoholic-yield yeast by HEPE and optimization of the fermentation condition

    International Nuclear Information System (INIS)

    Han Jingjing; Lu Jiangtao; Zhang Qin; Wang Yan; Fu Yujie; Wang Shilong; Fu Haiying

    2011-01-01

    The Saccharomyces Cerevisiae YE0 was mutated using high-energy-pulse-electron (HEPE) beam. After ethanol stress and determination of the alcohol yield by gas chromatograph, the mutant YF1 with high alcoholic yield was obtained. The results showed that under the optimized fermentation conditions (34 degree C as the fermentation temperature, 72 h as the fermentation time and 30% as the glucose concentration), the alcoholic yield of YF1 was 15.57% which was 58.23% higher than that of the original strain YE0 (9.84%) under the same conditions. The growth rate and lethal temperature of the mutant YF1 were obviously enhanced to the original strain YE0. The mutant YF1 has a great potential application in industrial production of alcohol. And it can also be used as the original strain for further mutagenesis to get the strain of higher alcoholic yield. (authors)

  8. Integration of ARTP mutagenesis with biosensor-mediated high-throughput screening to improve L-serine yield in Corynebacterium glutamicum.

    Science.gov (United States)

    Zhang, Xin; Zhang, Xiaomei; Xu, Guoqiang; Zhang, Xiaojuan; Shi, Jinsong; Xu, Zhenghong

    2018-05-03

    L-Serine is widely used in the pharmaceutical, food, and cosmetics industries. Although direct fermentative production of L-serine from sugar in Corynebacterium glutamicum has been achieved, the L-serine yield remains relatively low. In this study, atmospheric and room temperature plasma (ARTP) mutagenesis was used to improve the L-serine yield based on engineered C. glutamicum ΔSSAAI strain. Subsequently, we developed a novel high-throughput screening method using a biosensor constructed based on NCgl0581, a transcriptional factor specifically responsive to L-serine, so that L-serine concentration within single cell of C. glutamicum can be monitored via fluorescence-activated cell sorting (FACS). Novel L-serine-producing mutants were isolated from a large library of mutagenized cells. The mutant strain A36-pDser was screened from 1.2 × 10 5 cells, and the magnesium ion concentration in the medium was optimized specifically for this mutant. C. glutamicum A36-pDser accumulated 34.78 g/L L-serine with a yield of 0.35 g/g sucrose, which were 35.9 and 66.7% higher than those of the parent C. glutamicum ΔSSAAI-pDser strain, respectively. The L-serine yield achieved in this mutant was the highest of all reported L-serine-producing strains of C. glutamicum. Moreover, the whole-genome sequencing identified 11 non-synonymous mutations of genes associated with metabolic and transport pathways, which might be responsible for the higher L-serine production and better cell growth in C. glutamicum A36-pDser. This study explored an effective mutagenesis strategy and reported a novel high-throughput screening method for the development of L-serine-producing strains.

  9. Mutation-Guided Unbiased Modeling of the Fat Sensor GPR119 for High-Yield Agonist Screening

    DEFF Research Database (Denmark)

    Norn, Christoffer; Pedersen, Maria Hauge; Engelstoft, Maja S.

    2015-01-01

    and is consistent with the structure-activity relationship for a large no. of AR231453 analogs. Another key property of the refined models is their success in sepg. active ligands from decoys in a large-scale virtual screening. These results demonstrate that mutation-guided receptor modeling can provide predictions...

  10. DNA content analysis allows discrimination between Trypanosoma cruzi and Trypanosoma rangeli.

    Science.gov (United States)

    Naves, Lucila Langoni; da Silva, Marcos Vinícius; Fajardo, Emanuella Francisco; da Silva, Raíssa Bernardes; De Vito, Fernanda Bernadelli; Rodrigues, Virmondes; Lages-Silva, Eliane; Ramírez, Luis Eduardo; Pedrosa, André Luiz

    2017-01-01

    Trypanosoma cruzi, a human protozoan parasite, is the causative agent of Chagas disease. Currently the species is divided into six taxonomic groups. The genome of the CL Brener clone has been estimated to be 106.4-110.7 Mb, and DNA content analyses revealed that it is a diploid hybrid clone. Trypanosoma rangeli is a hemoflagellate that has the same reservoirs and vectors as T. cruzi; however, it is non-pathogenic to vertebrate hosts. The haploid genome of T. rangeli was previously estimated to be 24 Mb. The parasitic strains of T. rangeli are divided into KP1(+) and KP1(-). Thus, the objective of this study was to investigate the DNA content in different strains of T. cruzi and T. rangeli by flow cytometry. All T. cruzi and T. rangeli strains yielded cell cycle profiles with clearly identifiable G1-0 (2n) and G2-M (4n) peaks. T. cruzi and T. rangeli genome sizes were estimated using the clone CL Brener and the Leishmania major CC1 as reference cell lines because their genome sequences have been previously determined. The DNA content of T. cruzi strains ranged from 87,41 to 108,16 Mb, and the DNA content of T. rangeli strains ranged from 63,25 Mb to 68,66 Mb. No differences in DNA content were observed between KP1(+) and KP1(-) T. rangeli strains. Cultures containing mixtures of the epimastigote forms of T. cruzi and T. rangeli strains resulted in cell cycle profiles with distinct G1 peaks for strains of each species. These results demonstrate that DNA content analysis by flow cytometry is a reliable technique for discrimination between T. cruzi and T. rangeli isolated from different hosts.

  11. Trypanosoma evansi isolated from capybara (Hidrochaeris hidrochaeris

    Directory of Open Access Journals (Sweden)

    Karina Muñoz

    2001-10-01

    Full Text Available A study was conducted to determine the morphological and biometric characteristics of Trypanosoma isolated from 50 capybaras animals, raised in captivity in the Peruvian Amazon. Trypanosoma was found in 14 blood samples using the microhaematocrit, wide drop, and Giemsa-stain methods and T. evansi was identified through morphological details in all 14 positive samples (the subterminal kinetoplast, the developed undulating membrane, and a long free flagellum were used for the identification of the agent.

  12. Diverse inhibitor chemotypes targeting Trypanosoma cruzi CYP51.

    Directory of Open Access Journals (Sweden)

    Shamila S Gunatilleke

    Full Text Available Chagas Disease, a WHO- and NIH-designated neglected tropical disease, is endemic in Latin America and an emerging infection in North America and Europe as a result of population moves. Although a major cause of morbidity and mortality due to heart failure, as well as inflicting a heavy economic burden in affected regions, Chagas Disease elicits scant notice from the pharmaceutical industry because of adverse economic incentives. The discovery and development of new routes to chemotherapy for Chagas Disease is a clear priority.The similarity between the membrane sterol requirements of pathogenic fungi and those of the parasitic protozoon Trypanosoma cruzi, the causative agent of Chagas human cardiopathy, has led to repurposing anti-fungal azole inhibitors of sterol 14α-demethylase (CYP51 for the treatment of Chagas Disease. To diversify the therapeutic pipeline of anti-Chagasic drug candidates we exploited an approach that included directly probing the T. cruzi CYP51 active site with a library of synthetic small molecules. Target-based high-throughput screening reduced the library of ∼104,000 small molecules to 185 hits with estimated nanomolar K(D values, while cross-validation against T. cruzi-infected skeletal myoblast cells yielded 57 active hits with EC(50 <10 µM. Two pools of hits partially overlapped. The top hit inhibited T. cruzi with EC(50 of 17 nM and was trypanocidal at 40 nM.The hits are structurally diverse, demonstrating that CYP51 is a rather permissive enzyme target for small molecules. Cheminformatic analysis of the hits suggests that CYP51 pharmacology is similar to that of other cytochromes P450 therapeutic targets, including thromboxane synthase (CYP5, fatty acid ω-hydroxylases (CYP4, 17α-hydroxylase/17,20-lyase (CYP17 and aromatase (CYP19. Surprisingly, strong similarity is suggested to glutaminyl-peptide cyclotransferase, which is unrelated to CYP51 by sequence or structure. Lead compounds developed by pharmaceutical

  13. 21 CFR 866.3870 - Trypanosoma spp. serological reagents.

    Science.gov (United States)

    2010-04-01

    ... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3870 Trypanosoma... consist of antigens and antisera used in serological tests to identify antibodies to Trypanosoma spp. in...

  14. Characterization of plasma menbrane polypeptides of trypanosoma from bats

    OpenAIRE

    Pinho,R. T.; Simone,Giovanni de

    1989-01-01

    Cell surface proteins of Trypanosoma dionisii, Trypanosoma vespertilionis and Trypanosoma sp. (M238) were radiodinated and their distribution both in the detergent-poor (DPP) and dertergent-enriched phase (DRP) was studied using a phase separation technique in Triton X-114 as well as polyacrylamide gel electrophoresis in sodium dodecyl sulphate (SDS-PAGE). Significant differences were observed in the proteins present in the DRP when the three species of trypanosoma were compared. Two major ba...

  15. Screening Yield of HIV Antigen/Antibody Combination and Pooled HIV RNA Testing for Acute HIV Infection in a High-Prevalence Population.

    Science.gov (United States)

    Peters, Philip J; Westheimer, Emily; Cohen, Stephanie; Hightow-Weidman, Lisa B; Moss, Nicholas; Tsoi, Benjamin; Hall, Laura; Fann, Charles; Daskalakis, Demetre C; Beagle, Steve; Patel, Pragna; Radix, Asa; Foust, Evelyn; Kohn, Robert P; Marmorino, Jenni; Pandori, Mark; Fu, Jie; Samandari, Taraz; Gay, Cynthia L

    2016-02-16

    Although acute HIV infection contributes disproportionately to onward HIV transmission, HIV testing has not routinely included screening for acute HIV infection. To evaluate the performance of an HIV antigen/antibody (Ag/Ab) combination assay to detect acute HIV infection compared with pooled HIV RNA testing. Multisite, prospective, within-individual comparison study conducted between September 2011 and October 2013 in 7 sexually transmitted infection clinics and 5 community-based programs in New York, California, and North Carolina. Participants were 12 years or older and seeking HIV testing, without known HIV infection. All participants with a negative rapid HIV test result were screened for acute HIV infection with an HIV Ag/Ab combination assay (index test) and pooled human immunodeficiency virus 1 (HIV-1) RNA testing. HIV RNA testing was the reference standard, with positive reference standard result defined as detectable HIV-1 RNA on an individual RNA test. Number and proportion with acute HIV infections detected. Among 86,836 participants with complete test results (median age, 29 years; 75.0% men; 51.8% men who have sex with men), established HIV infection was diagnosed in 1158 participants (1.33%) and acute HIV infection was diagnosed in 168 participants (0.19%). Acute HIV infection was detected in 134 participants with HIV Ag/Ab combination testing (0.15% [95% CI, 0.13%-0.18%]; sensitivity, 79.8% [95% CI, 72.9%-85.6%]; specificity, 99.9% [95% CI, 99.9%-99.9%]; positive predictive value, 59.0% [95% CI, 52.3%-65.5%]) and in 164 participants with pooled HIV RNA testing (0.19% [95% CI, 0.16%-0.22%]; sensitivity, 97.6% [95% CI, 94.0%-99.4%]; specificity, 100% [95% CI, 100%-100%]; positive predictive value, 96.5% [95% CI, 92.5%-98.7%]; sensitivity comparison, P testing detected 82% of acute HIV infections detectable by pooled HIV RNA testing. Compared with rapid HIV testing alone, HIV Ag/Ab combination testing increased the relative HIV diagnostic yield (both

  16. Exploring the Trypanosoma brucei Hsp83 potential as a target for structure guided drug design.

    Directory of Open Access Journals (Sweden)

    Juan Carlos Pizarro

    Full Text Available Human African trypanosomiasis is a neglected parasitic disease that is fatal if untreated. The current drugs available to eliminate the causative agent Trypanosoma brucei have multiple liabilities, including toxicity, increasing problems due to treatment failure and limited efficacy. There are two approaches to discover novel antimicrobial drugs--whole-cell screening and target-based discovery. In the latter case, there is a need to identify and validate novel drug targets in Trypanosoma parasites. The heat shock proteins (Hsp, while best known as cancer targets with a number of drug candidates in clinical development, are a family of emerging targets for infectious diseases. In this paper, we report the exploration of T. brucei Hsp83--a homolog of human Hsp90--as a drug target using multiple biophysical and biochemical techniques. Our approach included the characterization of the chemical sensitivity of the parasitic chaperone against a library of known Hsp90 inhibitors by means of differential scanning fluorimetry (DSF. Several compounds identified by this screening procedure were further studied using isothermal titration calorimetry (ITC and X-ray crystallography, as well as tested in parasite growth inhibitions assays. These experiments led us to the identification of a benzamide derivative compound capable of interacting with TbHsp83 more strongly than with its human homologs and structural rationalization of this selectivity. The results highlight the opportunities created by subtle structural differences to develop new series of compounds to selectively target the Trypanosoma brucei chaperone and effectively kill the sleeping sickness parasite.

  17. Drug discovery for Chagas disease should consider Trypanosoma cruzi strain diversity

    Directory of Open Access Journals (Sweden)

    Bianca Zingales

    2014-09-01

    Full Text Available This opinion piece presents an approach to standardisation of an important aspect of Chagas disease drug discovery and development: selecting Trypanosoma cruzi strains for in vitro screening. We discuss the rationale for strain selection representing T. cruzi diversity and provide recommendations on the preferred parasite stage for drug discovery, T. cruzi discrete typing units to include in the panel of strains and the number of strains/clones for primary screens and lead compounds. We also consider experimental approaches for in vitro drug assays. The Figure illustrates the current Chagas disease drug-discovery and development landscape.

  18. Characterization of Trypanosoma brucei gambiense stocks isolated ...

    African Journals Online (AJOL)

    Trypanosoma brucei gambiense was isolated twice from each of 23 patients in Côte d'Ivoire. Genetic characterization using RAPD (Random Primed Amplified Polymorphic DNA) showed additional variability within a given isoenzyme profile (zymodeme), confirming that this fingerprinting method has a higher discriminative ...

  19. Colorectal cancer screening in patients with spinal cord injury yields similar results to the general population with an effective bowel preparation: a retrospective chart audit.

    Science.gov (United States)

    Teng, Brandon J; Song, Shawn H; Svircev, Jelena N; Dominitz, Jason A; Burns, Stephen P

    2018-03-01

    Retrospective chart audit. To compare adequacy of colonoscopy bowel preparation and diagnostic findings between persons with SCI receiving an extended inpatient bowel preparation and the general population. Veterans Affairs Puget Sound Healthcare System, Seattle, WA, USA. We reviewed an electronic database of all colonoscopies performed at a tertiary Veterans Affairs medical center between 7/12/13 and 15/10/15. Patients with SCI received a multi-day bowel preparation with magnesium citrate, and 8-10 liters of polyethylene glycol-3350 and electrolyte colonic lavage solution (PEG-ELS) over two and one half days. The control population received a standard bowel preparation consisting of magnesium citrate and 4 liters of PEG-ELS over 1 day. Two hundred and fifty-five patients were included in the study, including 85 patients with SCI. Average risk screening was a more common colonoscopy indication in patients with SCI vs. the control population (24 vs. 13% p = 0.03). There was no difference in adequacy of bowel preparation (87 vs. 85%, p = 0.73) or adenoma detection rate (55 vs. 51%, p = 0.59) when comparing patients with SCI with the control population. No difference in polyp histopathology was detected (p = 0.748). Our study demonstrated that an extended bowel preparation for patients with SCI produces similar bowel preparation results and diagnostic yield when compared to patients without SCI undergoing colonoscopy.

  20. Development and validation of polar RP-HPLC method for screening for ectoine high-yield strains in marine bacteria with green chemistry.

    Science.gov (United States)

    Chen, Jun; Chen, Jianwei; Wang, Sijia; Zhou, Guangmin; Chen, Danqing; Zhang, Huawei; Wang, Hong

    2018-04-02

    A novel, green, rapid, and precise polar RP-HPLC method has been successfully developed and screened for ectoine high-yield strain in marine bacteria. Ectoine is a polar and extremely useful solute which allows microorganisms to survive in extreme environmental salinity. This paper describes a polar-HPLC method employed polar RP-C18 (5 μm, 250 × 4.6 mm) using pure water as the mobile phase and a column temperature of 30 °C, coupled with a flow rate at 1.0 mL/min and detected under a UV detector at wavelength of 210 nm. Our method validation demonstrates excellent linearity (R 2  = 0.9993), accuracy (100.55%), and a limit of detection LOQ and LOD of 0.372 and 0.123 μgmL -1 , respectively. These results clearly indicate that the developed polar RP-HPLC method for the separation and determination of ectoine is superior to earlier protocols.

  1. Development of resazurin-based assay in 384-well format for high throughput whole cell screening of Trypanosoma brucei rhodesiense strain STIB 900 for the identification of potential anti-trypanosomal agents.

    Science.gov (United States)

    Lim, Kah Tee; Zahari, Zuriati; Amanah, Azimah; Zainuddin, Zafarina; Adenan, Mohd Ilham

    2016-03-01

    To accelerate the discovery of novel leads for the treatment of Human African Trypanosomiasis (HAT), it is necessary to have a simple, robust and cost-effective assay to identify positive hits by high throughput whole cell screening. Most of the fluorescence assay was made in black plate however in this study the HTS assay developed in 384-well format using clear plate and black plate, for comparison. The HTS assay developed is simple, sensitive, reliable and reproducible in both types of plates. Assay robustness and reproducibility were determined under the optimized conditions in 384-well plate was well tolerated in the HTS assay, including percentage of coefficient of variation (% CV) of 4.68% and 4.74% in clear and black 384-well plate, signal-to-background ratio (S/B) of 12.75 in clear 384-well plate and 12.07 in black 384-well plate, Z' factor of 0.79 and 0.82 in clear 384-well plate and black 384-well plate, respectively and final concentration of 0.30% dimethylsulfoxide (DMSO) in both types of plate. Drug sensitivity was found to be comparable to the reported anti-trypanosomal assay in 96-well format. The reproducibility and sensitivity of this assay make it compliant to automated liquid handler use in HTS applications. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Detection of Trypanosoma brucei gambiense and T. b. rhodesiense ...

    African Journals Online (AJOL)

    Detection of Trypanosoma brucei gambiense and T. b. rhodesiense in Glossina fuscipes fuscipes ( Diptera: Glossinidae ) and Stomoxys flies using the polymerase chain reaction (PCR) technique in southern Sudan.

  3. PCR-Based Detection of Trypanosoma evansi Infection in Semi-Captive Asiatic Black Bears (Ursus thibetanus

    Directory of Open Access Journals (Sweden)

    Maliha Shahid, Safia Janjua*, Fakhar-i-Abbas and Jan Schmidt Burbach1

    2013-11-01

    Full Text Available Clinical signs, viz lethargy, increased heart rate and reduced appetite, making trypanosomiasis a possible differential diagnosis, were found in five out of twenty semi-captive Asiatic black bears (Ursus thibetanus in a sanctuary, located in Kund, District Sawabi, KPK, Pakistan. Microscopic examination of blood samples of bears expressing clinical signs and symptoms revealed the presence of haemoflagellates, which was found to be trypanosomes. Subsequently, the PCR technique was exploited to screen for the presence of trypanosomal species in all bears’ blood samples. Blood samples from 20 individual bears were screened using three sets of primers specific to Trypanosoma evansi species. Three primer pairs used are equally effective in successful detection of the parasite. Two out of five, diseased bears died prior to any trypanosoma specific medication while the rest were given an administered dose of Melarsomine (Immiticide. The treated bears survived and were assured to be aparasitemic on post-treatment examination after six weeks.

  4. The population impact of human papillomavirus/cytology cervical cotesting at 3-year intervals: Reduced cervical cancer risk and decreased yield of precancer per screen.

    Science.gov (United States)

    Silver, Michelle I; Schiffman, Mark; Fetterman, Barbara; Poitras, Nancy E; Gage, Julia C; Wentzensen, Nicolas; Lorey, Thomas; Kinney, Walter K; Castle, Philip E

    2016-12-01

    The objective of cervical screening is to detect and treat precancer to prevent cervical cancer mortality and morbidity while minimizing overtreatment of benign human papillomavirus (HPV) infections and related minor abnormalities. HPV/cytology cotesting at extended 5-year intervals currently is a recommended screening strategy in the United States, but the interval extension is controversial. In the current study, the authors examined the impact of a decade of an alternative, 3-year cotesting, on rates of precancer and cancer at Kaiser Permanente Northern California. The effect on screening efficiency, defined as numbers of cotests/colposcopy visits needed to detect a precancer, also was considered. Two cohorts were defined. The "open cohort" included all women screened at least once during the study period; > 1 million cotests were performed. In a fixed "long-term screening cohort," the authors considered the cumulative impact of repeated screening at 3-year intervals by restricting the cohort to women first cotested in 2003 through 2004 (ie, no women entering screening later were added to this group). Detection of cervical intraepithelial neoplasia 3/adenocarcinoma in situ (CIN3/AIS) increased in the open cohort (2004-2006: 82.0/100,000 women screened; 2007-2009: 140.6/100,000 women screened; and 2010-2012: 126.0/100,000 women screened); cancer diagnoses were unchanged. In the long-term screening cohort, the detection of CIN3/AIS increased and then decreased to the original level (2004-2006: 80.5/100,000 women screened; 2007-2009: 118.6/100,000 women screened; and 2010-2012: 84.9./100,000 women screened). The number of cancer diagnoses was found to decrease. When viewed in terms of screening efficiency, the number of colposcopies performed to detect a single case of CIN3/AIS increased in the cohort with repeat screening. Repeated cotesting at a 3-year interval eventually lowers population rates of precancer and cancer. However, a greater number of

  5. The population impact of HPV/cytology cervical cotesting at 3-year intervals: reduced cervical cancer risk and decreased yield of precancer per screen

    Science.gov (United States)

    Silver, Michelle I.; Schiffman, Mark; Fetterman, Barbara; Poitras, Nancy; Gage, Julia C.; Wentzensen, Nicolas; Lorey, Thomas; Kinney, Walter; Castle, Philip E.

    2016-01-01

    Background Cervical screening aims to detect and treat precancer to prevent cervical cancer mortality and morbidity, while minimizing overtreatment of benign human papillomavirus (HPV) infections and related minor abnormalities. HPV/cytology cotesting at extended 5-year intervals is now a recommended screening strategy in the US, but the interval extension is controversial. We studied the impact of a decade of an alternative, 3-year cotesting, on rates of precancer and cancer at Kaiser Permanente Northern California. We also considered the effect on screening efficiency, defined as numbers of cotests/colposcopy visits needed to detect a precancer. Methods Two cohorts were defined. The “open cohort” included all women screened at least once during the study period; >1 million cotests were performed. In a fixed “long-term screening cohort”, we considered the cumulative impact of repeated screening at 3-year intervals by restricting to women first cotested in 2003–4 (i.e., no women entering screening later were added to this group). Results Detection of CIN3/AIS increased in the open cohort (2004–6, 82.0/100,000 women screened; 2007–9, 140.6/100,000; and 2010–12, 126.0/100,000); cancer diagnoses were unchanged. In the long-term screening cohort, detection of CIN3/AIS increased then decreased to the original level (2004–6, 80.5/100,000; 2007–9, 118.6/100,000; and 2010–2, 84.9/100,000). Cancer diagnoses decreased. Seen in terms of screening efficiency, the number of colposcopies performed todetect a single CIN3/AIS increased in the cohort with repeat screening. Conclusion Repeated cotesting at a 3-year interval eventually lowers population rates of precancer and cancer; however, a greater number of colposcopies is required to detect a single precancer. PMID:27657992

  6. Landscape epidemiology in urban environments: The example of rodent-borne Trypanosoma in Niamey, Niger.

    Science.gov (United States)

    Rossi, Jean-Pierre; Kadaouré, Ibrahima; Godefroid, Martin; Dobigny, Gauthier

    2017-10-05

    Trypanosomes are protozoan parasites found worldwide, infecting humans and animals. In the past decade, the number of reports on atypical human cases due to Trypanosoma lewisi or T. lewisi-like has increased urging to investigate the multiple factors driving the disease dynamics, particularly in cities where rodents and humans co-exist at high densities. In the present survey, we used a species distribution model, Maxent, to assess the spatial pattern of Trypanosoma-positive rodents in the city of Niamey. The explanatory variables were landscape metrics describing urban landscape composition and physiognomy computed from 8 land-cover classes. We computed the metrics around each data location using a set of circular buffers of increasing radii (20m, 40m, 60m, 80m and 100m). For each spatial resolution, we determined the optimal combination of feature class and regularization multipliers by fitting Maxent with the full dataset. Since our dataset was small (114 occurrences) we expected an important uncertainty associated to data partitioning into calibration and evaluation datasets. We thus performed 350 independent model runs with a training dataset representing a random subset of 80% of the occurrences and the optimal Maxent parameters. Each model yielded a map of habitat suitability over Niamey, which was transformed into a binary map implementing a threshold maximizing the sensitivity and the specificity. The resulting binary maps were combined to display the proportion of models that indicated a good environmental suitability for Trypanosoma-positive rodents. Maxent performed better with landscape metrics derived from buffers of 80m. Habitat suitability for Trypanosoma-positive rodents exhibited large patches linked to urban features such as patch richness and the proportion of landscape covered by concrete or tarred areas. Such inferences could be helpful in assessing areas at risk, setting of monitoring programs, public and medical staff awareness or even

  7. Rodent-borne Trypanosoma from cities and villages of Niger and Nigeria: A special role for the invasive genus Rattus?

    Science.gov (United States)

    Tatard, C; Garba, M; Gauthier, P; Hima, K; Artige, E; Dossou, D K H J; Gagaré, S; Genson, G; Truc, P; Dobigny, G

    2017-07-01

    Although they are known to sometimes infect humans, atypical trypanosomes are very poorly documented, especially in Africa where one lethal case has yet been described. Here we conducted a survey of rodent-borne Trypanosoma in 19 towns and villages of Niger and Nigeria, with a special emphasis on Niamey, the capital city of Niger. The 1298 rodents that were captured yielded 189 qPCR-positive animals from 14 localities, thus corresponding to a 14.6% overall prevalence. Rats, especially black rats, displayed particularly elevated prevalence (27.4%), with some well sampled sites showing 40-50% and up to 68.8% of Trypanosoma-carrying individuals. Rattus were also characterized by significantly lower Ct values than in the other non-Rattus species. DNA sequences could be obtained for 43 rodent-borne Trypanosoma and corresponded to 41 T. lewisi (all from Rattus) and 2 T. microti (from Cricetomys gambianus). These results, together with data compiled from the available literature, suggest that Rattus may play a particular role for the maintaining and circulation of Trypanosoma, especially T. lewisi, in Africa. Taken into account its strong abilities to invade coastal and inland regions of the continent, we believe that this genus deserves a particular attention in regards to potentially under-looked but emerging atypical trypanosome-related diseases. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Enzymatic Decoration of Prebiotic Galacto-oligosaccharides (Vivinal® GOS) with Sialic Acid using Trypanosoma cruzi Trans-Sialidase and Two Bovine Sialoglycoconjugates as Donor Substrates

    NARCIS (Netherlands)

    Wilbrink, Maarten Hotse; Ten Kate, Geert Albert; Sanders, Peter; Gerwig, Gerrit J; van Leeuwen, Sander S; Sallomons, Erik; Klarenbeek, Bert; Hage, Johannes H; van Vuure, Carine A; Dijkhuizen, Lubbert; Kamerling, Johannis P

    2015-01-01

    Decoration of prebiotic galacto-oligosaccharides (GOS) with sialic acid yields mixtures of GOS and sialylated GOS (Sia-GOS), novel products that are expected to have both prebiotic and anti-adhesive functionalities. The recombinantly produced trans-sialidase enzyme from Trypanosoma cruzi (TcTS), an

  9. Trypanosoma brucei gambiense trypanosomiasis in Terego county, northern Uganda, 1996: a lot quality assurance sampling survey.

    Science.gov (United States)

    Hutin, Yvan J F; Legros, Dominique; Owini, Vincent; Brown, Vincent; Lee, Evan; Mbulamberi, Dawson; Paquet, Christophe

    2004-04-01

    We estimated the pre-intervention prevalence of Trypanosoma brucei gambiense (Tbg) trypanosomiasis using the lot quality assurance sampling (LQAS) methods in 14 parishes of Terego County in northern Uganda. A total of 826 participants were included in the survey sample in 1996. The prevalence of laboratory confirmed Tbg trypanosomiasis adjusted for parish population sizes was 2.2% (95% confidence interval =1.1-3.2). This estimate was consistent with the 1.1% period prevalence calculated on the basis of cases identified through passive and active screening in 1996-1999. Ranking of parishes in four categories according to LQAS analysis of the 1996 survey predicted the prevalences observed during the first round of active screening in the population in 1997-1998 (P LQAS were validated by the results of the population screening, suggesting that these survey methods may be useful in the pre-intervention phase of sleeping sickness control programs.

  10. Congenital Trypanosoma cruzi Transmission in Santa Cruz, Bolivia

    Science.gov (United States)

    Bern, Caryn; Verastegui, Manuela; Gilman, Robert H.; LaFuente, Carlos; Galdos-Cardenas, Gerson; Calderon, Maritza; Pacori, Juan; Abastoflor, Maria del Carmen; Aparicio, Hugo; Brady, Mark F.; Ferrufino, Lisbeth; Angulo, Noelia; Marcus, Sarah; Sterling, Charles; Maguire, James H.

    2017-01-01

    Background We conducted a study of congenital Trypanosoma cruzi infection in Santa Cruz, Bolivia. Our objective was to apply new tools to identify weak points in current screening algorithms, and find ways to improve them. Methods Women presenting for delivery were screened by rapid and conventional serological tests. For infants of infected mothers, blood specimens obtained on days 0, 7, 21, 30, 90, 180, and 270 were concentrated and examined microscopically; serological tests were performed for the day 90, 180, and 270 specimens. Maternal and infant specimens, including umbilical tissue, were tested by polymerase chain reaction (PCR) targeting the kinetoplast minicircle and by quantitative PCR. Results Of 530 women, 154 (29%) were seropositive. Ten infants had congenital T. cruzi infection. Only 4 infants had positive results of microscopy evaluation in the first month, and none had positive cord blood microscopy results. PCR results were positive for 6 (67%) of 9 cord blood and 7 (87.5%) of 8 umbilical tissue specimens. PCR-positive women were more likely to transmit T. cruzi than were seropositive women with negative PCR results (P < .05). Parasite loads determined by quantitative PCR were higher for mothers of infected infants than for seropositive mothers of uninfected infants (P < .01). Despite intensive efforts, only 58% of at-risk infants had a month 9 specimen collected. Conclusions On the basis of the low sensitivity of microscopy in cord blood and high rate of loss to follow-up, we estimate that current screening programs miss one-half of all infected infants. Molecular techniques may improve early detection. PMID:19877966

  11. Secondary Metabolites from Vietnamese Marine Invertebrates with Activity against Trypanosoma brucei and T. cruzi

    Directory of Open Access Journals (Sweden)

    Nguyen Phuong Thao

    2014-06-01

    Full Text Available Marine-derived natural products from invertebrates comprise an extremely diverse and promising source of the compounds from a wide variety of structural classes. This study describes the discovery of five marine natural products with activity against Trypanosoma species by natural product library screening using whole cell in vitro assays. We investigated the anti-trypanosomal activity of the extracts from the soft corals and echinoderms living in Vietnamese seas. Of the samples screened, the methanolic extracts of several marine organisms exhibited potent activities against cultures of Trypanosoma brucei and T. cruzi (EC50 < 5.0 μg/mL. Among the compounds isolated from these extracts, laevigatol B (1 from Lobophytum crassum and L. laevigatum, (24S-ergost-4-ene-3-one (2 from Sinularia dissecta, astropectenol A (3 from Astropecten polyacanthus, and cholest-8-ene-3β,5α,6β,7α-tetraol (4 from Diadema savignyi showed inhibitory activity against T. brucei with EC50 values ranging from 1.57 ± 0.14 to 14.6 ± 1.36 μM, relative to the positive control, pentamidine (EC50 = 0.015 ± 0.003 μM. Laevigatol B (1 and 5α-cholest-8(14-ene-3β,7α-diol (5 exhibited also significant inhibitory effects on T. cruzi. The cytotoxic activity of the pure compounds on mammalian cells was also assessed and found to be insignificant in all cases. This is the first report on the inhibitory effects of marine organisms collected in Vietnamese seas against Trypanosoma species responsible for neglected tropical diseases.

  12. Iron-associated biology of Trypanosoma brucei.

    Czech Academy of Sciences Publication Activity Database

    Basu, Somsuvro; Horáková, Eva; Lukeš, Julius

    2016-01-01

    Roč. 1860, č. 2 (2016), s. 363-370 ISSN 0304-4165 R&D Projects: GA ČR(CZ) GA14-23986S; GA ČR GAP305/12/2261; GA MŠk(CZ) EE2.3.30.0032 EU Projects: European Commission(XE) COST Action CM1307; European Commission(XE) 316304 - MODBIOLIN Grant - others:AV ČR(CZ) M200961204 Institutional support: RVO:60077344 Keywords : iron * Fe/S cluster * heme * Trypanosoma * TAO Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.702, year: 2016

  13. [Trypanosoma cruzi in triatomines from Nuevo Leon, Mexico].

    Science.gov (United States)

    Molina-Garza, Zinnia Judith; Rosales-Encina, José Luis; Galaviz-Silva, Lucio; Molina-Garza, Daniel

    2007-01-01

    To determine the prevalence of Trypanosoma cruzi in triatomines from Nuevo León using the standardization of an improved enzyme-linked immunosorbent assay test. From July to September 2005, 52 triatomines were captured in General Terán, a municipality located in Nuevo León. They were analyzed using optical microscopy (OM) and a polymerase chain reaction (PCR), as standards of reference, to develop a technique for detecting the parasite using enzyme-linked immunosorbent assay (ELISA). Using OM and PCR, 31 triatomines were found to be positive and 21 negative. Using ELISA, 27 samples were identified as positive and 25 negative (specificity 100%, sensitivity 87%, negative predictive value 84%, and positive predictive value 100%). The prevalence of infected triatomines was 59.61% with OM and PCR, and 51.92% with ELISA. Our data confirm that the ELISA assay in triatomines is a fast, reliable and useful tool. Since it was possible to simultaneously analyze a large number of samples with high sensibility and specificity values, the ELISA test proves to be useful for new epidemiologic studies having a high number of vectors. It is also less expensive than PCR. It is therefore recommended for epidemiological and preventive surveillance programs as a first screening test before conducting a confirmatory test using PCR.

  14. Anti-Trypanosoma cruzi antibody detection in eastern Andalusia (Spain).

    Science.gov (United States)

    Marín, Clotilde; Concha-Valdez, Fanny; Cañas, Rocío; Gutiérrez-Sánchez, Ramón; Sánchez-Moreno, Manuel

    2014-03-01

    Chagas disease caused by the protozoan haemoflagellate Trypanosoma cruzi is no longer found exclusively in Latin America; the disease is occurring in Europe, and Spain is the country with the highest prevalence. Our aim was to detect anti-T. cruzi antibodies in blood donors from southeast Spain, and we performed eight serological diagnostic assays on each of 550 blood samples collected in March-June 2010. Two in-house ELISA methods were used to test against a parasite lysate (ELISA-H) and the semi-purified superoxide dismutase excreted by T. cruzi (ELISA-SODe); we also used the Western blot technique against the same antigen (WB-SODe), indirect immunofluorescence (IFA) and four commercial tests. The serological test results showed a range of seroprevalence values, the lowest being 1.1%, determined by IFA and two commercial tests (Ab rapid and Chagascreen); other values were: 1.3% (commercial ELISA [Chagas ELISA IgG+IgM]); 2.1% (immunochromatographic test [Stick Chagas]); 2.7% (ELISA-H); 4.0% (WB-SODe); and 4.2%, the highest value (ELISA-SODe). The excellent specificity of SODe antigen for the detection of antibodies to T. cruzi in donors lead us to affirm that the serological test performed with this biomarker could provide a useful screening and confirmatory test method for cases of Chagas disease.

  15. Prevalence of Trypanosoma cruzi/HIV coinfection in southern Brazil

    Directory of Open Access Journals (Sweden)

    Dulce Stauffert

    2017-03-01

    Full Text Available Chagas disease reactivation has been a defining condition for acquired immune deficiency syndrome in Brazil for individuals coinfected with Trypanosoma cruzi and HIV since 2004. Although the first coinfection case was reported in the 1980s, its prevalence has not been firmly established. In order to know coinfection prevalence, a cross-sectional study of 200 HIV patients was performed between January and July 2013 in the city of Pelotas, in southern Rio Grande do Sul, an endemic area for Chagas disease. Ten subjects were found positive for T. cruzi infection by chemiluminescence microparticle immunoassay and indirect immunofluorescence. The survey showed 5% coinfection prevalence among HIV patients (95% CI: 2.0–8.0, which was 3.8 times as high as that estimated by the Ministry of Health of Brazil. Six individuals had a viral load higher than 100,000 copies per μL, a statistically significant difference for T. cruzi presence. These findings highlight the importance of screening HIV patients from Chagas disease endemic areas.

  16. Phenolic Constituents of Medicinal Plants with Activity against Trypanosoma brucei

    Directory of Open Access Journals (Sweden)

    Ya Nan Sun

    2016-04-01

    Full Text Available Neglected tropical diseases (NTDs affect over one billion people all over the world. These diseases are classified as neglected because they impact populations in areas with poor financial conditions and hence do not attract sufficient research investment. Human African Trypanosomiasis (HAT or sleeping sickness, caused by the parasite Trypanosoma brucei, is one of the NTDs. The current therapeutic interventions for T. brucei infections often have toxic side effects or require hospitalization so that they are not available in the rural environments where HAT occurs. Furthermore, parasite resistance is increasing, so that there is an urgent need to identify novel lead compounds against this infection. Recognizing the wide structural diversity of natural products, we desired to explore and identify novel antitrypanosomal chemotypes from a collection of natural products obtained from plants. In this study, 440 pure compounds from various medicinal plants were tested against T. brucei by in a screening using whole cell in vitro assays. As the result, twenty-two phenolic compounds exhibited potent activity against cultures of T. brucei. Among them, eight compounds—4, 7, 11, 14, 15, 18, 20, and 21—showed inhibitory activity against T. brucei, with IC50 values below 5 µM, ranging from 0.52 to 4.70 μM. Based on these results, we attempt to establish some general trends with respect to structure-activity relationships, which indicate that further investigation and optimization of these derivatives might enable the preparation of potentially useful compounds for treating HAT.

  17. Molecular recognition and self-assembly special feature: A general protocol for creating high-throughput screening assays for reaction yield and enantiomeric excess applied to hydrobenzoin.

    Science.gov (United States)

    Shabbir, Shagufta H; Regan, Clinton J; Anslyn, Eric V

    2009-06-30

    A general approach to high-throughput screening of enantiomeric excess (ee) and concentration was developed by using indicator displacement assays (IDAs), and the protocol was then applied to the vicinal diol hydrobenzoin. The method involves the sequential utilization of what we define herein as screening, training, and analysis plates. Several enantioselective boronic acid-based receptors were screened by using 96-well plates, both for their ability to discriminate the enantiomers of hydrobenzoin and to find their optimal pairing with indicators resulting in the largest optical responses. The best receptor/indicator combination was then used to train an artificial neural network to determine concentration and ee. To prove the practicality of the developed protocol, analysis plates were created containing true unknown samples of hydrobenzoin generated by established Sharpless asymmetric dihydroxylation reactions, and the best ligand was correctly identified.

  18. Species-specific markers for the differential diagnosis of Trypanosoma cruzi and Trypanosoma rangeli and polymorphisms detection in Trypanosoma rangeli.

    Science.gov (United States)

    Ferreira, Keila Adriana Magalhães; Fajardo, Emanuella Francisco; Baptista, Rodrigo P; Macedo, Andrea Mara; Lages-Silva, Eliane; Ramírez, Luis Eduardo; Pedrosa, André Luiz

    2014-06-01

    Trypanosoma cruzi and Trypanosoma rangeli are kinetoplastid parasites which are able to infect humans in Central and South America. Misdiagnosis between these trypanosomes can be avoided by targeting barcoding sequences or genes of each organism. This work aims to analyze the feasibility of using species-specific markers for identification of intraspecific polymorphisms and as target for diagnostic methods by PCR. Accordingly, primers which are able to specifically detect T. cruzi or T. rangeli genomic DNA were characterized. The use of intergenic regions, generally divergent in the trypanosomatids, and the serine carboxypeptidase gene were successful. Using T. rangeli genomic sequences for the identification of group-specific polymorphisms and a polymorphic AT(n) dinucleotide repeat permitted the classification of the strains into two groups, which are entirely coincident with T. rangeli main lineages, KP1 (+) and KP1 (-), previously determined by kinetoplast DNA (kDNA) characterization. The sequences analyzed totalize 622 bp (382 bp represent a hypothetical protein sequence, and 240 bp represent an anonymous sequence), and of these, 581 (93.3%) are conserved sites and 41 bp (6.7%) are polymorphic, with 9 transitions (21.9%), 2 transversions (4.9%), and 30 (73.2%) insertion/deletion events. Taken together, the species-specific markers analyzed may be useful for the development of new strategies for the accurate diagnosis of infections. Furthermore, the identification of T. rangeli polymorphisms has a direct impact in the understanding of the population structure of this parasite.

  19. Repurposing a Library of Human Cathepsin L Ligands: Identification of Macrocyclic Lactams as Potent Rhodesain and Trypanosoma brucei Inhibitors.

    Science.gov (United States)

    Giroud, Maude; Dietzel, Uwe; Anselm, Lilli; Banner, David; Kuglstatter, Andreas; Benz, Jörg; Blanc, Jean-Baptiste; Gaufreteau, Delphine; Liu, Haixia; Lin, Xianfeng; Stich, August; Kuhn, Bernd; Schuler, Franz; Kaiser, Marcel; Brun, Reto; Schirmeister, Tanja; Kisker, Caroline; Diederich, François; Haap, Wolfgang

    2018-04-26

    Rhodesain (RD) is a parasitic, human cathepsin L (hCatL) like cysteine protease produced by Trypanosoma brucei ( T. b.) species and a potential drug target for the treatment of human African trypanosomiasis (HAT). A library of hCatL inhibitors was screened, and macrocyclic lactams were identified as potent RD inhibitors ( K i < 10 nM), preventing the cell-growth of Trypanosoma brucei rhodesiense (IC 50 < 400 nM). SARs addressing the S2 and S3 pockets of RD were established. Three cocrystal structures with RD revealed a noncovalent binding mode of this ligand class due to oxidation of the catalytic Cys25 to a sulfenic acid (Cys-SOH) during crystallization. The P-glycoprotein efflux ratio was measured and the in vivo brain penetration in rats determined. When tested in vivo in acute HAT model, the compounds permitted up to 16.25 (vs 13.0 for untreated controls) mean days of survival.

  20. 3-H-[1,2]Dithiole as a New Anti-Trypanosoma cruzi Chemotype: Biological and Mechanism of Action Studies

    Directory of Open Access Journals (Sweden)

    Marcos Couto

    2015-08-01

    Full Text Available The current pharmacological Chagas disease treatments, using Nifurtimox or Benznidazole, show limited therapeutic results and are associated with potential side effects, like mutagenicity. Using random screening we have identified new chemotypes that were able to inhibit relevant targets of the Trypanosoma cruzi. We found 3H-[1,2]dithioles with the ability to inhibit Trypanosoma cruzi triosephosphate isomerase (TcTIM. Herein, we studied the structural modifications of this chemotype to analyze the influence of volume, lipophilicity and electronic properties in the anti-T. cruzi activity. Their selectivity to parasites vs. mammalian cells was also examined. To get insights into a possible mechanism of action, the inhibition of the enzymatic activity of TcTIM and cruzipain, using the isolated enzymes, and the inhibition of membrane sterol biosynthesis and excreted metabolites, using the whole parasite, were achieved. We found that this structural framework is interesting for the generation of innovative drugs for the treatment of Chagas disease.

  1. Vaccination with Trypanosoma rangeli induces resistance of guinea pigs to virulent Trypanosoma cruzi.

    Science.gov (United States)

    Basso, B; Moretti, E; Fretes, R

    2014-01-15

    Chagas' disease, endemic in Latin America, is spread in natural environments through animal reservoirs, including marsupials, mice and guinea pigs. Farms breeding guinea pigs for food are located in some Latin-American countries with consequent risk of digestive infection. The aim of this work was to study the effect of vaccination with Trypanosoma rangeli in guinea pigs challenged with Trypanosoma cruzi. Animals were vaccinated with fixated epimastigotes of T. rangeli, emulsified with saponin. Controls received only PBS. Before being challenged with T. cruzi, parasitemia, survival rates and histological studies were performed. The vaccinated guinea pigs revealed significantly lower parasitemia than controls (pguinea pigs and dogs. The development of vaccines for use in animals, like domestic dogs and guinea pigs in captivity, opens up new opportunities for preventive tools, and could reduce the risk of infection with T. cruzi in the community. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Indices to screen for grain yield and grain-zinc mass concentrations in aerobic rice at different soil-Zn levels

    NARCIS (Netherlands)

    Jiang, W.; Struik, P.C.; Zhao, M.; Keulen, van H.; Fan, T.Q.; Stomph, T.J.

    2008-01-01

    Zinc is an important micronutrient for both crop growth and human nutrition. In rice production, yields are often reduced and Zn mass concentrations in the grains are often low when Zn is in short supply to the crop. This may result in malnutrition of people dependent on a rice-based diet. Plant

  3. Classical clinical signs in rats experimemtally infected with Trypanosoma brucei

    Directory of Open Access Journals (Sweden)

    Nwoha Rosemary Ijeoma Ogechi

    2015-02-01

    Full Text Available Objective: To investigate clinical signs in Trypanosoma brucei infection in albino rats. Methods: Fourteen rats grouped into 2 with 7 rats in each group were used to determine classical clinical manifestation of Trypanosoma brucei infection in rats. Group A rats were uninfected control and Group B rats were infected with Trypanosoma brucei. Results: Parasitaemia was recorded in Group B by (3.86±0.34 d and the peak of parasitaemia was observed at Day 5 post infection. Classical signs observed included squint eyes, raised whiskers, lethargy, no weight loss, pyrexia, isolation from the other rats, and starry hair coat. Conclusions: These signs could be diagnostic or aid in diagnosis of Trypanosoma brucei infection in rats.

  4. Role of cytokines in Trypanosoma brucei-induced anaemia: A ...

    African Journals Online (AJOL)

    species Trypanosoma brucei that are transmitted by a tsetse fly (Glossina spp.) ... of autologous immunoglobulin antibodies on the red cell surfaces and also to ... development for the detection and management of anaemia in trypanosomiasis.

  5. Quantitative Proteomic and Phosphoproteomic Analysis of Trypanosoma cruzi Amastigogenesis

    DEFF Research Database (Denmark)

    Queiroz, Rayner M L; Charneau, Sebastien; Mandacaru, Samuel C

    2014-01-01

    Chagas disease is a tropical neglected disease endemic in Latin America and it is caused by the protozoan Trypanosoma cruzi. The parasite has four major life stages: epimastigote, metacyclic trypomastigote, bloodstream trypomastigote and amastigote. The differentiation from infective trypomastigo......Chagas disease is a tropical neglected disease endemic in Latin America and it is caused by the protozoan Trypanosoma cruzi. The parasite has four major life stages: epimastigote, metacyclic trypomastigote, bloodstream trypomastigote and amastigote. The differentiation from infective...

  6. Equity yields

    NARCIS (Netherlands)

    Vrugt, E.; van Binsbergen, J.H.; Koijen, R.S.J.; Hueskes, W.

    2013-01-01

    We study a new data set of dividend futures with maturities up to ten years across three world regions: the US, Europe, and Japan. We use these asset prices to construct equity yields, analogous to bond yields. We decompose the equity yields to obtain a term structure of expected dividend growth

  7. Molecular Diversity of Trypanosoma cruzi Detected in the Vector Triatoma protracta from California, USA.

    Directory of Open Access Journals (Sweden)

    Lisa A Shender

    2016-01-01

    Full Text Available Trypanosoma cruzi, causative agent of Chagas disease in humans and dogs, is a vector-borne zoonotic protozoan parasite that can cause fatal cardiac disease. While recognized as the most economically important parasitic infection in Latin America, the incidence of Chagas disease in the United States of America (US may be underreported and even increasing. The extensive genetic diversity of T. cruzi in Latin America is well-documented and likely influences disease progression, severity and treatment efficacy; however, little is known regarding T. cruzi strains endemic to the US. It is therefore important to expand our knowledge on US T. cruzi strains, to improve upon the recognition of and response to locally acquired infections.We conducted a study of T. cruzi molecular diversity in California, augmenting sparse genetic data from southern California and for the first time investigating genetic sequences from northern California. The vector Triatoma protracta was collected from southern (Escondido and Los Angeles and northern (Vallecito California regions. Samples were initially screened via sensitive nuclear repetitive DNA and kinetoplast minicircle DNA PCR assays, yielding an overall prevalence of approximately 28% and 55% for southern and northern California regions, respectively. Positive samples were further processed to identify discrete typing units (DTUs, revealing both TcI and TcIV lineages in southern California, but only TcI in northern California. Phylogenetic analyses (targeting COII-ND1, TR and RB19 genes were performed on a subset of positive samples to compare Californian T. cruzi samples to strains from other US regions and Latin America. Results indicated that within the TcI DTU, California sequences were similar to those from the southeastern US, as well as to several isolates from Latin America responsible for causing Chagas disease in humans.Triatoma protracta populations in California are frequently infected with T. cruzi

  8. Screening and Identification of putative long non coding RNAs from transcriptome data of a high yielding blackgram (Vigna mungo, Cv. T9

    Directory of Open Access Journals (Sweden)

    Pankaj Kumar Singh

    2018-04-01

    Full Text Available Blackgram (Vigna mungo is one of primary legumes cultivated throughout India, Cv.T9 being one of its common high yielding cultivar. This article reports RNA sequencing data and a pipeline for prediction of novel long non-coding RNAs from the sequenced data. The raw data generated during sequencing are available at Sequence Read Archive (SRA of NCBI with accession number- SRX1558530 Keywords: Blackgram, Long non-coding RNA, Legumes, RNA sequencing data

  9. Stearoyl-CoA desaturase is an essential enzyme for the parasitic protist Trypanosoma brucei

    Energy Technology Data Exchange (ETDEWEB)

    Alloatti, Andres [Instituto de Biologia Molecular y Celular de Rosario (IBR), CONICET, Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario, Santa Fe (Argentina); Gupta, Shreedhara; Gualdron-Lopez, Melisa; Nguewa, Paul A. [Research Unit for Tropical Diseases, de Duve Institute and Laboratory of Biochemistry, Universite Catholique de Louvain, Brussels (Belgium); Altabe, Silvia G. [Instituto de Biologia Molecular y Celular de Rosario (IBR), CONICET, Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario, Santa Fe (Argentina); Deumer, Gladys; Wallemacq, Pierre [Department of Clinical Chemistry, Cliniques Universitaires Saint-Luc, LTAP, Universite Catholique de Louvain, Brussels (Belgium); Michels, Paul A.M. [Research Unit for Tropical Diseases, de Duve Institute and Laboratory of Biochemistry, Universite Catholique de Louvain, Brussels (Belgium); Uttaro, Antonio D., E-mail: toniuttaro@yahoo.com.ar [Instituto de Biologia Molecular y Celular de Rosario (IBR), CONICET, Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario, Santa Fe (Argentina)

    2011-08-26

    Highlights: {yields} Inhibiting {Delta}9 desaturase drastically changes T. brucei's fatty-acid composition. {yields} Isoxyl specifically inhibits the {Delta}9 desaturase causing a growth arrest. {yields} RNA interference of desaturase expression causes a similar effect. {yields} Feeding T. brucei-infected mice with Isoxyl decreases the parasitemia. {yields} 70% of Isoxyl-treated mice survived the trypanosome infection. -- Abstract: Trypanosoma brucei, the etiologic agent of sleeping sickness, is exposed to important changes in nutrients and temperature during its life cycle. To adapt to these changes, the fluidity of its membranes plays a crucial role. This fluidity, mediated by the fatty-acid composition, is regulated by enzymes named desaturases. We have previously shown that the oleoyl desaturase is essential for Trypanosoma cruzi and T. brucei. In this work, we present experimental support for the relevance of stearoyl-CoA desaturase (SCD) for T. brucei's survival, in both its insect or procyclic-form (PCF) and bloodstream-form (BSF) stages. We evaluated this essentiality in two different ways: by generating a SCD knocked-down parasite line using RNA interference, and by chemical inhibition of the enzyme with two compounds, Isoxyl and a thiastearate with the sulfur atom at position 10 (10-TS). The effective concentration for 50% growth inhibition (EC{sub 50}) of PCF was 1.0 {+-} 0.2 {mu}M for Isoxyl and 5 {+-} 2 {mu}M for 10-TS, whereas BSF appeared more susceptible with EC{sub 50} values 0.10 {+-} 0.03 {mu}M (Isoxyl) and 1.0 {+-} 0.6 {mu}M (10-TS). RNA interference showed to be deleterious for both stages of the parasite. In addition, T. brucei-infected mice were fed with Isoxyl, causing a reduction of the parasitemia and an increase of the rodents' survival.

  10. The haemoculture of Trypanosoma minasense chagas, 1908

    Directory of Open Access Journals (Sweden)

    Mariangela Ziccardi

    1996-08-01

    Full Text Available Trypanosoma minasense was isolated for the first time in blood axenic culture from a naturally infected marmoset, Callithrix penicillata, from Brazil. The parasite grew profusely in an overlay of Roswell Park Memorial Institute medium plus 20% foetal bovine serum, on Novy, McNeal and Nicolle medium (NNN , at 27°C, with a peak around 168 hr. The morphometry of cultural forms of T. minasense, estimates of cell population size and comparative growth in four different media overlays always with NNN, were studied. The infectivity of cultural forms to marmosets (C. penicillata and C. jacchus and transformation of epimastigotes into metacyclic-like forms in axenic culture in the presence of chitin derivates (chitosan were evaluated.

  11. Proteomics of Trypanosoma evansi infection in rodents.

    Science.gov (United States)

    Roy, Nainita; Nageshan, Rishi Kumar; Pallavi, Rani; Chakravarthy, Harshini; Chandran, Syama; Kumar, Rajender; Gupta, Ashok Kumar; Singh, Raj Kumar; Yadav, Suresh Chandra; Tatu, Utpal

    2010-03-22

    Trypanosoma evansi infections, commonly called 'surra', cause significant economic losses to livestock industry. While this infection is mainly restricted to large animals such as camels, donkeys and equines, recent reports indicate their ability to infect humans. There are no World Animal Health Organization (WAHO) prescribed diagnostic tests or vaccines available against this disease and the available drugs show significant toxicity. There is an urgent need to develop improved methods of diagnosis and control measures for this disease. Unlike its related human parasites T. brucei and T. cruzi whose genomes have been fully sequenced T. evansi genome sequence remains unavailable and very little efforts are being made to develop improved methods of prevention, diagnosis and treatment. With a view to identify potential diagnostic markers and drug targets we have studied the clinical proteome of T. evansi infection using mass spectrometry (MS). Using shot-gun proteomic approach involving nano-lc Quadrupole Time Of Flight (QTOF) mass spectrometry we have identified over 160 proteins expressed by T. evansi in mice infected with camel isolate. Homology driven searches for protein identification from MS/MS data led to most of the matches arising from related Trypanosoma species. Proteins identified belonged to various functional categories including metabolic enzymes; DNA metabolism; transcription; translation as well as cell-cell communication and signal transduction. TCA cycle enzymes were strikingly missing, possibly suggesting their low abundances. The clinical proteome revealed the presence of known and potential drug targets such as oligopeptidases, kinases, cysteine proteases and more. Previous proteomic studies on Trypanosomal infections, including human parasites T. brucei and T. cruzi, have been carried out from lab grown cultures. For T. evansi infection this is indeed the first ever proteomic study reported thus far. In addition to providing a glimpse into the

  12. Proteomics of Trypanosoma evansi infection in rodents.

    Directory of Open Access Journals (Sweden)

    Nainita Roy

    2010-03-01

    Full Text Available Trypanosoma evansi infections, commonly called 'surra', cause significant economic losses to livestock industry. While this infection is mainly restricted to large animals such as camels, donkeys and equines, recent reports indicate their ability to infect humans. There are no World Animal Health Organization (WAHO prescribed diagnostic tests or vaccines available against this disease and the available drugs show significant toxicity. There is an urgent need to develop improved methods of diagnosis and control measures for this disease. Unlike its related human parasites T. brucei and T. cruzi whose genomes have been fully sequenced T. evansi genome sequence remains unavailable and very little efforts are being made to develop improved methods of prevention, diagnosis and treatment. With a view to identify potential diagnostic markers and drug targets we have studied the clinical proteome of T. evansi infection using mass spectrometry (MS.Using shot-gun proteomic approach involving nano-lc Quadrupole Time Of Flight (QTOF mass spectrometry we have identified over 160 proteins expressed by T. evansi in mice infected with camel isolate. Homology driven searches for protein identification from MS/MS data led to most of the matches arising from related Trypanosoma species. Proteins identified belonged to various functional categories including metabolic enzymes; DNA metabolism; transcription; translation as well as cell-cell communication and signal transduction. TCA cycle enzymes were strikingly missing, possibly suggesting their low abundances. The clinical proteome revealed the presence of known and potential drug targets such as oligopeptidases, kinases, cysteine proteases and more.Previous proteomic studies on Trypanosomal infections, including human parasites T. brucei and T. cruzi, have been carried out from lab grown cultures. For T. evansi infection this is indeed the first ever proteomic study reported thus far. In addition to providing a

  13. Use of Nonspecific, Glutamic Acid-Free, Media and High Glycerol or High Amylase as Inducing Parameters for Screening Bacillus Isolates Having High Yield of Polyglutamic Acid.

    Science.gov (United States)

    Baxi, Nandita N

    2014-01-01

    Out of fifty-five Bacillus isolates obtained from ten different regional locations and sources, seven showed the ability to consistently produce specific extracellular polymeric substance (EPS) on rich as well as synthetic but nonspecific media which did not contain glutamic acid. The isolates were identified as either Bacillus licheniformis or Bacillus subtilis. The EPS from all isolates was resistant to alpha protease, proteinase K, and was thus of high molecular weight. Further it was detected after SDS-PAGE by methylene blue but not by coomassie blue R staining as in case of proteins with high proportion of acidic amino acids. Cell-free EPS, after acid hydrolysis, showed absence of carbohydrates and presence of only glutamic acid. Thus the native the EPS from all seven isolates was confirmed to be gamma polyglutamic acid (PGA) and not exopolysaccharide. The Bacillus isolate T which produced maximum polymer on all media tested had higher amylase: protease activity as compared to other strains. If inoculum was developed in rich medium as compared to synthetic medium, the PGA produced increased by twofold in the subsequent synthetic production medium. Similarly, use of inoculum consisting of young and vegetative cells also increased the PGA production by twofold though amount of inoculum did not affect yield of PGA. Though PGA was produced in even in the absence of glutamic acid supplementation in the production medium by all isolates, the yield of PGA increased by fourfold in the presence glutamic acid and the maximum yield was 30 g/l for isolate K. The supplementation of glutamine instead of glutamic acid into the medium caused an increase in the viscosity of the non-Newtonian solution of PGA.

  14. Phytochemical Screening and In Vitro Antitrypanosomal Activity of ...

    African Journals Online (AJOL)

    Phytochemical Screening and In Vitro Antitrypanosomal Activity of the Aerial Parts of Artemisia abyssinica Against Trypanosoma congolense Field Isolate. ... of infected blood and extracts at concentrations of 4, 2 and 0.4 mg/ml coupled with infectivity test in which a mixture of infected blood was inoculated to healthy mice.

  15. Seroprevalence of Trypanosoma cruzi in blood donors at the National Blood Transfusion Services--Guyana.

    Science.gov (United States)

    Bwititi, P T; Browne, J

    2012-09-01

    Blood transfusion is an important transmission route of Trypanosoma cruzi (T cruzi), a major parasitic infection in Central and South America. The limited treatment options are most effective in acute Chagas' infection. At present, there is no current data on the prevalence of T cruzi in the blood donor population of Guyana. This information is necessary to protect the supply of the blood donation programme. This study sought to determine the prevalence of T cruzi in the blood supply at the National Blood Transfusion Services of Guyana with the hope of providing knowledge to the on-going surveillance for Chagas' disease worldwide and therefore address the risk of its spread by blood transfusion. Two commercialized ELISAs utilizing crude or recombinant T cruzi antigens were used to study 2000 blood samples voluntarily donated for the purpose of altruistic or family replacement donation retrospectively. The results showed that approximately 1 in 286 donations tested positive for antibodies to T cruzi. These results indicate that T cruzi continues to be a risk in Guyana and there is a need to continue screening donated blood. Trypanosoma cruzi is a life-long infection and infected persons may be asymptomatic chronic carriers of the disease. Education, housing improvement, and controlled use of insecticides should be introduced to contain Chagas' disease.

  16. Evaluation of In Vitro Activity of Essential Oils against Trypanosoma brucei brucei and Trypanosoma evansi

    Directory of Open Access Journals (Sweden)

    Nathan Habila

    2010-01-01

    Full Text Available Essential oils (EOs from Cymbopogon citratus (CC, Eucalyptus citriodora (EC, Eucalyptus camaldulensis (ED, and Citrus sinensis (CS were obtained by hydrodistillation process. The EOs were evaluated in vitro for activity against Trypanosoma brucei brucei (Tbb and Trypanosoma evansi (T. evansi. The EOs were found to possess antitrypanosomal activity in vitro in a dose-dependent pattern in a short period of time. The drop in number of parasite over time was achieved doses of 0.4 g/ml, 0.2 g/mL, and 0.1 g/mL for all the EOs. The concentration of 0.4 g/mL CC was more potent at 3 minutes and 2 minutes for Tbb and T. evansi, respectively. The GC-MS analysis of the EOs revealed presence of Cyclobutane (96.09% in CS, 6-octenal (77.11% in EC, Eucalyptol (75% in ED, and Citral (38.32% in CC among several other organic compounds. The results are discussed in relation to trypanosome chemotherapy.

  17. Molecular diagnosis of cattle trypanosomes in Venezuela: evidences of Trypanosoma evansi and Trypanosoma vivax infections.

    Science.gov (United States)

    Ramírez-Iglesias, J R; Eleizalde, M C; Reyna-Bello, A; Mendoza, M

    2017-06-01

    In South America Trypanosoma evansi has been determined by molecular methods in cattle from Bolivia, Brazil, Colombia and Peru, reason for which the presence of this parasite is not excluded in Venezuelan livestock. Therefore, the aim of this study was to perform parasitological and molecular diagnosis of cattle trypanosomosis in small livestock units from two regions in this country. The parasitological diagnosis was carried out by MHCT and the molecular by PCR using genus-specific ITS1 primers that differentiate T. vivax and T. evansi infections. 47 cattle were evaluated in the "Laguneta de la Montaña" sector, Miranda State, where 3 animals were diagnosed as positive (6.4 %) by MHCT and 14 (30 %) by PCR as Trypanosoma spp., out of which 9 animals resulted positive for T. vivax , 3 for T. evansi and 2 with double infections. Whilst in the "San Casimiro" sector, State of Aragua, out of the 38 cattle evaluated 7 animals were diagnosed as positive (18.4 %) by MHCT and 19 (50 %) by PCR, determining only the presence of T. evansi in this locality. The molecular diagnosis by PCR using ITS1 primers allowed T. evansi detection in cattle field populations, which suggests the possible role of these animals as reservoirs in the epidemiology of the disease caused by T. evansi in Venezuela.

  18. Troglitazone induces differentiation in Trypanosoma brucei

    International Nuclear Information System (INIS)

    Denninger, Viola; Figarella, Katherine; Schoenfeld, Caroline; Brems, Stefanie; Busold, Christian; Lang, Florian; Hoheisel, Joerg; Duszenko, Michael

    2007-01-01

    Trypanosoma brucei, a protozoan parasite causing sleeping sickness, is transmitted by the tsetse fly and undergoes a complex lifecycle including several defined stages within the insect vector and its mammalian host. In the latter, differentiation from the long slender to the short stumpy form is induced by a yet unknown factor of trypanosomal origin. Here we describe that some thiazolidinediones are also able to induce differentiation. In higher eukaryotes, thiazolidinediones are involved in metabolism and differentiation processes mainly by binding to the intracellular receptor peroxisome proliferator activated receptor γ. Our studies focus on the effects of troglitazone on bloodstream form trypanosomes. Differentiation was monitored using mitochondrial markers (membrane potential, succinate dehydrogenase activity, inhibition of oxygen uptake by KCN, amount of cytochrome transcripts), morphological changes (Transmission EM and light microscopy), and transformation experiments (loss of the Variant Surface Glycoprotein coat and increase of dihydroliponamide dehydrogenase activity). To further investigate the mechanisms responsible for these changes, microarray analyses were performed, showing an upregulation of expression site associated gene 8 (ESAG8), a potential differentiation regulator

  19. Gastrointestinal parasites and Trypanosoma evansi in buffaloes

    International Nuclear Information System (INIS)

    Sani, R.A.; Chandrawathani, P.; Rosli, M.

    1990-01-01

    Gastrointestinal parasitism is common in buffalo calves. The effect of helminths on growth was studied by administration of an anthelmintic to buffalo calves following natural infections with gastrointestinal parasites. In studies conducted on calves belonging to an institute and a smallholder farmer, the treated calves showed improved weight gains. Serial parasitic examinations showed these animals had moderate to high faecal counts with Strongyloides, Toxocara vitulorum and Haemonchus eggs and Eimeria oocytes. In another study, there was no live weight advantage in treated over untreated calves. Few animals in this study had evidence of parasites and even those which were infested had low faecal egg counts. Hence, in general, helminths at certain levels of infection do affect the live weight gains of young buffalo calves. The prevalence of Trypanosoma evansi, as assessed parasitologically using the haematocrit centrifugation technique and mice inoculation, was 2.7 and 1%, respectively, in cattle and buffaloes. The serological prevalence using the enzyme linked immunosorbent assay was 35 and 2% for cattle and buffaloes, respectively. (author). 6 refs, 5 figs, 2 tabs

  20. Cell signaling during Trypanosoma cruzi invasion

    Directory of Open Access Journals (Sweden)

    Fernando Yukio Maeda

    2012-11-01

    Full Text Available Cell signaling is an essential requirement for mammalian cell invasion by Trypanosoma cruzi. Depending on the parasite strain and the parasite developmental form, distinct signaling pathways may be induced. In this short review, we focus on the data coming from studies with metacyclic trypomastigotes (MT generated in vitro and tissue culture-derived trypomastigotes (TCT, used as counterparts of insect-borne and bloodstream parasites respectively. During invasion of host cells by MT or TCT, intracellular Ca2+ mobilization and host cell lysosomal exocytosis are triggered. Invasion mediated by MT surface molecule gp82 requires the activation of mammalian target of rapamycin (mTOR, phosphatidylinositol 3-kinase (PI3K and protein kinase C (PKC in the host cell, associated with Ca2+-dependent disruption of the actin cytoskeleton. In MT, protein tyrosine kinase (PTK, PI3K, phospholipase C (PLC and PKC appear to be activated. TCT invasion, on the other hand, does not rely on mTOR activation, rather on target cell PI3K, and may involve the host cell autophagy for parasite internalization. Enzymes, such oligopeptidase B and the major T. cruzi cysteine proteinase cruzipain, have been shown to generate molecules that induce target cell Ca2+ signal. In addition, TCT may trigger host cell responses mediated by TGF-β receptor or integrin family member. Further investigations are needed for a more complete and detailed picture of T. cruzi invasion.

  1. Flagellar Motility of Trypanosoma cruzi Epimastigotes

    Directory of Open Access Journals (Sweden)

    G. Ballesteros-Rodea

    2012-01-01

    Full Text Available The hemoflagellate Trypanosoma cruzi is the causative agent of American trypanosomiasis. Despite the importance of motility in the parasite life cycle, little is known about T. cruzi motility, and there is no quantitative description of its flagellar beating. Using video microscopy and quantitative vectorial analysis of epimastigote trajectories, we find a forward parasite motility defined by tip-to-base symmetrical flagellar beats. This motion is occasionally interrupted by base-to-tip highly asymmetric beats, which represent the ciliary beat of trypanosomatid flagella. The switch between flagellar and ciliary beating facilitates the parasite's reorientation, which produces a large variability of movement and trajectories that results in different distance ranges traveled by the cells. An analysis of the distance, speed, and rotational angle indicates that epimastigote movement is not completely random, and the phenomenon is highly dependent on the parasite behavior and is characterized by directed and tumbling parasite motion as well as their combination, resulting in the alternation of rectilinear and intricate motility paths.

  2. Differential gene expression during Trypanosoma cruzi metacyclogenesis

    Directory of Open Access Journals (Sweden)

    Marco Aurelio Krieger

    1999-09-01

    Full Text Available The transformation of epimastigotes into metacyclic trypomastigotes involves changes in the pattern of expressed genes, resulting in important morphological and functional differences between these developmental forms of Trypanosoma cruzi. In order to identify and characterize genes involved in triggering the metacyclogenesis process and in conferring to metacyclic trypomastigotes their stage specific biological properties, we have developed a method allowing the isolation of genes specifically expressed when comparing two close related cell populations (representation of differential expression or RDE. The method is based on the PCR amplification of gene sequences selected by hybridizing and subtracting the populations in such a way that after some cycles of hybridization-amplification genes specific to a given population are highly enriched. The use of this method in the analysis of differential gene expression during T. cruzi metacyclogenesis (6 hr and 24 hr of differentiation and metacyclic trypomastigotes resulted in the isolation of several clones from each time point. Northern blot analysis showed that some genes are transiently expressed (6 hr and 24 hr differentiating cells, while others are present in differentiating cells and in metacyclic trypomastigotes. Nucleotide sequencing of six clones characterized so far showed that they do not display any homology to gene sequences available in the GeneBank.

  3. Studies on the glycosome of Trypanosoma brucei

    International Nuclear Information System (INIS)

    Aman, R.A.

    1985-01-01

    Glycosomes (microbodies) have been purified from bloodstream form Trypanosoma brucei by an improved procedure involving freezing and thawing live organisms in 15% glycerol prior to cell disruption. Highly purified organelles of bloodstream form T. brucei contain 11 major proteins of which 8 tentatively identified glycolytic enzymes make up about 90% of the total glycosomal protein. Treatment of these intact isolated organelles with the bisimidoester dimethylsuberimidate (DMSI) resulted in crosslinking of all glycosomal proteins into a large complex suggestive of juxtapositioning of the glycosomal proteins. The crosslinked complex was capable of catalyzing the multienzyme conversion of glucose to glycerol-3-phosphate but did not possess any special kinetic features different from those of the unaggregated enzymes represented by solubilized glycosomes. The multienzyme reaction had a lab phase associated with it and [ 14 C]-glucose label incorporation into sugar phosphate intermediates was effectively competed by unlabeled intermediates. Glycosomes were also purified from culture form T. brucei by several different procedures. Comparison of highly purified organelles from the two different life stages of the organism showed reduced specific activities and contents of the early glycolytic enzymes in organelles from the culture form with a decrease from 87% to 35% of the contribution of glycolytic enzymes to the total glycosomal protein

  4. Variant surface glycoproteins from Venezuelan trypanosome isolates are recognized by sera from animals infected with either Trypanosoma evansi or Trypanosoma vivax.

    Science.gov (United States)

    Camargo, Rocío; Izquier, Adriana; Uzcanga, Graciela L; Perrone, Trina; Acosta-Serrano, Alvaro; Carrasquel, Liomary; Arias, Laura P; Escalona, José L; Cardozo, Vanessa; Bubis, José

    2015-01-15

    Salivarian trypanosomes sequentially express only one variant surface glycoprotein (VSG) on their cell surface from a large repertoire of VSG genes. Seven cryopreserved animal trypanosome isolates known as TeAp-ElFrio01, TEVA1 (or TeAp-N/D1), TeGu-N/D1, TeAp-Mantecal01, TeGu-TerecayTrino, TeGu-Terecay03 and TeGu-Terecay323, which had been isolated from different hosts identified in several geographical areas of Venezuela were expanded using adult albino rats. Soluble forms of predominant VSGs expressed during the early infection stages were purified and corresponded to concanavalin A-binding proteins with molecular masses of 48-67 kDa by sodium dodecyl sulfate-polyacrylamide gel electropohoresis, and pI values between 6.1 and 7.5. The biochemical characterization of all purified soluble VSGs revealed that they were dimers in their native form and represented different gene products. Sequencing of some of these proteins yielded peptides homologous to VSGs from Trypanosoma (Trypanozoon) brucei and Trypanosoma (Trypanozoon) evansi and established that they most likely are mosaics generated by homologous recombination. Western blot analysis showed that all purified VSGs were cross-reacting antigens that were recognized by sera from animals infected with either T. evansi or Trypanosoma (Dutonella) vivax. The VSG glycosyl-phosphatidylinositol cross-reacting determinant epitope was only partially responsible for the cross-reactivity of the purified proteins, and antibodies appeared to recognize cross-reacting conformational epitopes from the various soluble VSGs. ELISA experiments were performed using infected bovine sera collected from cattle in a Venezuelan trypanosome-endemic area. In particular, soluble VSGs from two trypanosome isolates, TeGu-N/D1 and TeGu-TeracayTrino, were recognized by 93.38% and 73.55% of naturally T. vivax-infected bovine sera, respectively. However, approximately 70% of the sera samples did not recognize all seven purified proteins. Hence, the

  5. A Highly Sensitive Rapid Diagnostic Test for Chagas Disease That Utilizes a Recombinant Trypanosoma cruzi Antigen

    Science.gov (United States)

    Barfield, C. A.; Barney, R. S.; Crudder, C. H.; Wilmoth, J. L.; Stevens, D. S.; Mora-Garcia, S.; Yanovsky, M. J.; Weigl, B. H.; Yanovsky, J.

    2011-01-01

    Improved diagnostic tests for Chagas disease are urgently needed. A new lateral flow rapid test for Chagas disease is under development at PATH, in collaboration with Laboratorio Lemos of Argentina, which utilizes a recombinant antigen for detection of antibodies to Trypanosoma cruzi. To evaluate the performance of this test, 375 earlier characterized serum specimens from a region where Chagas is endemic were tested using a reference test (the Ortho T. cruzi ELISA, Johnson & Johnson), a commercially available rapid test (Chagas STAT-PAK, Chembio), and the PATH–Lemos rapid test. Compared to the composite reference tests, the PATH–Lemos rapid test demonstrated an optimal sensitivity of 99.5% and specificity of 96.8%, while the Chagas STAT-PAK demonstrated a sensitivity of 95.3% and specificity of 99.5%. These results indicate that the PATH–Lemos rapid test shows promise as an improved and reliable tool for screening and diagnosis of Chagas disease. PMID:21342808

  6. Experimental methods for screening parameters influencing the growth to product yield (Y(x/CH4 of a biological methane production (BMP process performed with Methanothermobacter marburgensis

    Directory of Open Access Journals (Sweden)

    Sébastien Bernacchi

    2014-12-01

    simple, unstructured mathematical model and experimental verification with Methanobacterium thermoautotrophicum. Biotechnol Bioeng 51: 645-658.12. Peillex JP, Fardeau ML, Belaich JP (1990 Growth of Methanobacterium thermoautotrophicum on hydrogen-carbon dioxide: high methane productivities in continuous culture. Biomass 21:315-321.13. Nishimura N, Kitaura S, Mimura A, et al. (1992 Cultivation of thermophilic methanogen KN-15 on hydrogen-carbon dioxide under pressurized conditions. J Ferment Bioeng 73: 477-480.14. Morii H, Koga Y, Nagai S (1987 Energetic analysis of the growth of Methanobrevibacter arboriphilus A2 in hydrogen-limited continuous cultures. Biotechnol Bioeng 29: 310-315.15. de Poorter LMI, Geerts WJ, Keltjens JT (2007 Coupling of Methanothermobacter thermautotrophicus methane formation and growth in fed-batch and continuous cultures under different H2 gassing regimens. Appl Environ Microbiol 73: 740-749.16. Schoenheit P, Moll J, Thauer RK (1980 Growth parameters (Ks, μmax, Ys of Methanobacterium thermoautotrophicum. Arch Microbiol 127: 59-65.17. Gerhard E, Butsch BM, Marison IW, et al. (1993 Improved growth and methane production conditions for Methanobacterium thermoautotrophicum. Appl Microbiol Biotechnol 40: 432-437.18. Seifert AH, Rittmann S, Bernacchi S, et al. (2013 Method for assessing the impact of emission gasses on physiology and productivity in biological methanogenesis. Bioresour Technol 136:747-751.19. Fardeau ML, Peillex JP, Belaich JP (1987 Energetics of the growth of Methanobacterium thermoautotrophicum and Methanococcus thermolithotrophicus on ammonium chloride and dinitrogen. Arch Microbiol 148: 128-131.20. Fardeau ML, Belaich JP (1986 Energetics of the growth of Methanococcus thermolithotrophicus. Arch Microbiol 144: 381-385.21. Morgan RM, Pihl TD, Nolling J (1997 Hydrogen regulation of growth, growth yields, and methane gene transcription in Methanobacterium thermoautotrophicum Delta H. J Bacteriol 179:889-898.22. Archer DB (1985

  7. First record of Trypanosoma chattoni in Brazil and occurrence of other Trypanosoma species in Brazilian frogs (Anura, Leptodactylidae).

    Science.gov (United States)

    Lemos, M; Morais, D H; Carvalho, V T; D'Agosto, M

    2008-02-01

    The present study provides the first record of Trypanosoma chattoni Mathis and Leger, 1911, in a new host, Leptodactylus fuscus Schneider, 1799 (Anura, Leptodactylidae), and the occurrence of Trypanosoma rotatorium-like species in Leptodactylus chaquensis Cei, 1950. The anurans were captured in the State of Mato Grosso, Brazil. Blood samples were obtained by cardiac puncture, and blood smears were examined for the presence of hemoparasites. The Trypanosoma rotatorium-like species in this study refers to a short-bodied trypomastigote that has a conspicuous undulating membrane but lacks a free flagellum; T. chattoni refers to a monomorphic parasite that has a rounded body, a kinetoplast adjacent to the nucleus, and a short flagellum.

  8. EPIDEMIOLOGÍA MOLECULAR DE TRYPANOSOMA CRUZI

    Directory of Open Access Journals (Sweden)

    Felipe Guhl

    2013-01-01

    Full Text Available La enfermedad de Chagas causada por el parásito Trypanosoma cruzi es una zoonosis compleja, ampliamente distribuida en el continente americano. La infección puede ser adquirida a través de las heces de insectos triatominos, transfusión de sangre, trasplante de órganos, vía oral, por transmisión congénita y por accidentes de laboratorio. El completo entendimiento de la etiología y epidemiología de la enfermedad de Chagas a través de su distribución geográfica es complejo y permanece bajo intensa investigación hasta la actualidad. Los recientes estudios sobre la variabilidad genética del parásito han dado nuevas luces de los diferentes escenarios de los ciclos de transmisión de la enfermedad y su patogénesis en humanos. El propósito principal para la caracterización molecular de T.cruzi y sus múltiples genotipos está dirigido hacia su asociación con la clínica y la patogenesis de la enfermedad, así como al esclarecimiento de los diferentes escenarios de transmisión y los aspectos coevolutivos relacionados con reservorios e insectos vectores. La caracterización molecular de los diferentes aislamientos a partir de humanos, insectos y reservorios, ha permitido identificar la amplia variabilidad genética del parásito, abriendo nuevos caminos hacia la búsqueda de nuevos blancos terapéuticos y pruebas diagnósticas más específicas que contribuyan a mitigar la enfermedad de Chagas.

  9. Trypanosoma brucei mitochondrial respiratome: Composition and organization in procyclic form

    KAUST Repository

    Acestor, Nathalie

    2011-05-24

    The mitochondrial respiratory chain is comprised of four different protein complexes (I-IV), which are responsible for electron transport and generation of proton gradient in the mitochondrial intermembrane space. This proton gradient is then used by F oF 1-ATP synthase (complex V) to produce ATP by oxidative phosphorylation. In this study, the respiratory complexes I, II, and III were affinity purified from Trypanosoma brucei procyclic form cells and their composition was determined by mass spectrometry. The results along with those that we previously reported for complexes IV and V showed that the respiratome of Trypanosoma is divergent because many of its proteins are unique to this group of organisms. The studies also identified two mitochondrial subunit proteins of respiratory complex IV that are encoded by edited RNAs. Proteomics data from analyses of complexes purified using numerous tagged component proteins in each of the five complexes were used to generate the first predicted protein-protein interaction network of the Trypanosoma brucei respiratory chain. These results provide the first comprehensive insight into the unique composition of the respiratory complexes in Trypanosoma brucei, an early diverged eukaryotic pathogen. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. A tropical tale: how Naja nigricollis venom beats Trypanosoma brucei

    DEFF Research Database (Denmark)

    Martos Esteban, Andrea; Laustsen, Andreas Hougaard; Carrington, Mark

    Trypanosoma brucei is a parasitic protozoan species capable to infecting insect vectors whose bite further produces African sleeping sickness inhuman beings [1]. During the parasite’s extracellular life in the mammalian host,its outer coat, mainly composed of Variable Surface Glycoproteins (VSGs)...

  11. What controls glycolysis in bloodstream form Trypanosoma brucei?

    NARCIS (Netherlands)

    Bakker, B.M.; Michels, P.A.M.; Opperdoes, F.R.; Westerhoff, H.V.

    1999-01-01

    On the basis of the experimentally determined kinetic properties of the trypanosomal enzymes, the question is addressed of which step limits the glycolytic flux in bloodstream form Trypanosoma brucei. There appeared to be no single answer; in the physiological range, control shifted between the

  12. Trypanosoma cruzi: avirulence of the PF strain to Callithrix marmosets

    Directory of Open Access Journals (Sweden)

    Humberto Menezes

    1981-06-01

    Full Text Available Callithrix jacchus geoffroy marmosets (HumBol. 1812 were injected once subcutaneously with 10.000 parasites/g body weight and followed for a period of six months. The PF strain of Trypanosoma cruzi was used. Follow-up was done through blood cultures, xenodiagnosis, serological tests, and ECG. A small number of normaI animais served as control.

  13. Role of sialic acids in the midguts of Trypanosoma congolense ...

    African Journals Online (AJOL)

    Administrator

    total sialic acid concentration. The relevance of these findings to the role of sialic acids in the midgut of. T. congolense infected C.p. pipiense mosquitoes is discussed in this paper. Key words: Trypanosoma congolense, Culex pipiense pipiense, sialic acid, midgut. INTRODUCTION. The Culex pipiense pipiense mosquito is ...

  14. Serum total protein, albumin and globulin levels in Trypanosoma ...

    African Journals Online (AJOL)

    The effect of orally administered Scoparia dulcis on Trypanosoma brucei-induced changes in serum total protein, albumin and globulin were investigated in rabbits over a period of twenty eight days. Results obtained show that infection resulted in hyperproteinaemia, hyperglobulinaemia and hypoalbuminaemia. However ...

  15. Targeting the HSP60/10 chaperonin systems of Trypanosoma brucei as a strategy for treating African sleeping sickness.

    Science.gov (United States)

    Abdeen, Sanofar; Salim, Nilshad; Mammadova, Najiba; Summers, Corey M; Goldsmith-Pestana, Karen; McMahon-Pratt, Diane; Schultz, Peter G; Horwich, Arthur L; Chapman, Eli; Johnson, Steven M

    2016-11-01

    Trypanosoma brucei are protozoan parasites that cause African sleeping sickness in humans (also known as Human African Trypanosomiasis-HAT). Without treatment, T. brucei infections are fatal. There is an urgent need for new therapeutic strategies as current drugs are toxic, have complex treatment regimens, and are becoming less effective owing to rising antibiotic resistance in parasites. We hypothesize that targeting the HSP60/10 chaperonin systems in T. brucei is a viable anti-trypanosomal strategy as parasites rely on these stress response elements for their development and survival. We recently discovered several hundred inhibitors of the prototypical HSP60/10 chaperonin system from Escherichia coli, termed GroEL/ES. One of the most potent GroEL/ES inhibitors we discovered was compound 1. While examining the PubChem database, we found that a related analog, 2e-p, exhibited cytotoxicity to Leishmania major promastigotes, which are trypanosomatids highly related to Trypanosoma brucei. Through initial counter-screening, we found that compounds 1 and 2e-p were also cytotoxic to Trypanosoma brucei parasites (EC 50 =7.9 and 3.1μM, respectively). These encouraging initial results prompted us to develop a library of inhibitor analogs and examine their anti-parasitic potential in vitro. Of the 49 new chaperonin inhibitors developed, 39% exhibit greater cytotoxicity to T. brucei parasites than parent compound 1. While many analogs exhibit moderate cytotoxicity to human liver and kidney cells, we identified molecular substructures to pursue for further medicinal chemistry optimization to increase the therapeutic windows of this novel class of chaperonin-targeting anti-parasitic candidates. An intriguing finding from this study is that suramin, the first-line drug for treating early stage T. brucei infections, is also a potent inhibitor of GroEL/ES and HSP60/10 chaperonin systems. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Comparative analysis of the kinomes of three pathogenic trypanosomatids: Leishmania major, Trypanosoma brucei and Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Ward Pauline N

    2005-09-01

    Full Text Available Abstract Background The trypanosomatids Leishmania major, Trypanosoma brucei and Trypanosoma cruzi cause some of the most debilitating diseases of humankind: cutaneous leishmaniasis, African sleeping sickness, and Chagas disease. These protozoa possess complex life cycles that involve development in mammalian and insect hosts, and a tightly coordinated cell cycle ensures propagation of the highly polarized cells. However, the ways in which the parasites respond to their environment and coordinate intracellular processes are poorly understood. As a part of an effort to understand parasite signaling functions, we report the results of a genome-wide analysis of protein kinases (PKs of these three trypanosomatids. Results Bioinformatic searches of the trypanosomatid genomes for eukaryotic PKs (ePKs and atypical PKs (aPKs revealed a total of 176 PKs in T. brucei, 190 in T. cruzi and 199 in L. major, most of which are orthologous across the three species. This is approximately 30% of the number in the human host and double that of the malaria parasite, Plasmodium falciparum. The representation of various groups of ePKs differs significantly as compared to humans: trypanosomatids lack receptor-linked tyrosine and tyrosine kinase-like kinases, although they do possess dual-specificity kinases. A relative expansion of the CMGC, STE and NEK groups has occurred. A large number of unique ePKs show no strong affinity to any known group. The trypanosomatids possess few ePKs with predicted transmembrane domains, suggesting that receptor ePKs are rare. Accessory Pfam domains, which are frequently present in human ePKs, are uncommon in trypanosomatid ePKs. Conclusion Trypanosomatids possess a large set of PKs, comprising approximately 2% of each genome, suggesting a key role for phosphorylation in parasite biology. Whilst it was possible to place most of the trypanosomatid ePKs into the seven established groups using bioinformatic analyses, it has not been

  17. Fragment-based screening in tandem with phenotypic screening provides novel antiparasitic hits.

    Science.gov (United States)

    Blaazer, Antoni R; Orrling, Kristina M; Shanmugham, Anitha; Jansen, Chimed; Maes, Louis; Edink, Ewald; Sterk, Geert Jan; Siderius, Marco; England, Paul; Bailey, David; de Esch, Iwan J P; Leurs, Rob

    2015-01-01

    Methods to discover biologically active small molecules include target-based and phenotypic screening approaches. One of the main difficulties in drug discovery is elucidating and exploiting the relationship between drug activity at the protein target and disease modification, a phenotypic endpoint. Fragment-based drug discovery is a target-based approach that typically involves the screening of a relatively small number of fragment-like (molecular weight <300) molecules that efficiently cover chemical space. Here, we report a fragment screening on TbrPDEB1, an essential cyclic nucleotide phosphodiesterase (PDE) from Trypanosoma brucei, and human PDE4D, an off-target, in a workflow in which fragment hits and a series of close analogs are subsequently screened for antiparasitic activity in a phenotypic panel. The phenotypic panel contained T. brucei, Trypanosoma cruzi, Leishmania infantum, and Plasmodium falciparum, the causative agents of human African trypanosomiasis (sleeping sickness), Chagas disease, leishmaniasis, and malaria, respectively, as well as MRC-5 human lung cells. This hybrid screening workflow has resulted in the discovery of various benzhydryl ethers with antiprotozoal activity and low toxicity, representing interesting starting points for further antiparasitic optimization. © 2014 Society for Laboratory Automation and Screening.

  18. Trypanocidal activity of Brazilian plants against epimastigote forms from Y and Bolivia strains of Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Renata Tomé Alves

    2012-01-01

    Full Text Available Chagas disease is one of the main public health problems in Latin America. Since the available treatments for this disease are not effective in providing cure, the screening of potential antiprotozoal agents is essential, mainly of those obtained from natural sources. This study aimed to provide an evaluation of the trypanocidal activity of 92 ethanol extracts from species belonging to the families Annonaceae, Apiaceae, Cucurbitaceae, Lamiaceae, Lauraceae, Moraceae, Nyctaginaceae, and Verbenaceae against the Y and Bolivia strains of Trypanosoma cruzi. Additionally, cytotoxic activity on LLCMK2 fibroblasts was evaluated. Both the trypanocidal activity and cytotoxicity were evaluated using the MTT method, in the following concentrations: 500, 350, 250, and 100 µg/mL. Benznidazole was used for positive control. The best results among the 92 samples evaluated were obtained with ethanol extracts of Ocotea paranapiacabensis (Am93 and Aegiphila lhotzkiana (Am160. Am93 showed trypanocidal activity against epimastigote forms of the Bolivia strain and was moderately toxic to LLCMK2 cells, its Selectivity Index (SI being 14.56, while Am160 showed moderate trypanocidal activity against the Bolivia strain and moderate toxicicity, its SI being equal to 1.15. The screening of Brazilian plants has indicated the potential effect of ethanol extracts obtained from Ocotea paranapiacabensis and Aegiphila lhotzkiana against Chagas disease.

  19. Meiosis and Haploid Gametes in the Pathogen Trypanosoma brucei

    OpenAIRE

    Peacock, Lori; Bailey, Mick; Carrington, Mark; Gibson, Wendy

    2014-01-01

    Summary In eukaryote pathogens, sex is an important driving force in spreading genes for drug resistance, pathogenicity, and virulence [1]. For the parasitic trypanosomes that cause African sleeping sickness, mating occurs during transmission by the tsetse vector [2, 3] and involves meiosis [4], but haploid gametes have not yet been identified. Here, we show that meiosis is a normal part of development in the insect salivary glands for all subspecies of Trypanosoma brucei, including the human...

  20. Trypanosoma brucei Mitochondrial Respiratome: Composition and Organization in Procyclic Form

    Czech Academy of Sciences Publication Activity Database

    Acestor, N.; Zíková, Alena; Dalley, R. A.; Anupama, A.; Panigrahi, A. K.; Stuart, K. D.

    2011-01-01

    Roč. 10, č. 9 (2011), s. 1-14 ISSN 1535-9476 R&D Projects: GA ČR GP204/09/P563 Institutional research plan: CEZ:AV0Z60220518 Keywords : SUCCINATE DEHYDROGENASE * EDITED MESSENGER-RNA * COMPLEX-I * TRYPANOSOMA-BRUCEI * UBIQUINONE OXIDOREDUCTASE * TAP-TAG * PROTEIN INTERACTION * ALTERNATIVE OXIDASE * STATISTICAL-MODEL * MASS-SPECTROMETRY Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 7.398, year: 2011

  1. Trypanosoma brucei solanesyl-diphosphate synthase localizes to the mitochondrion

    Czech Academy of Sciences Publication Activity Database

    Lai, D.-H.; Bontempi, E. J.; Lukeš, Julius

    2012-01-01

    Roč. 183, č. 2 (2012), s. 189-192 ISSN 0166-6851 R&D Projects: GA ČR(CZ) GAP305/11/2179 Institutional support: RVO:60077344 Keywords : Trypanosoma brucei * Sleeping sickness * Ubiquinone * Solanesyl-diphosphate synthase * Digitonin permeabilization * In situ tagging Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.734, year: 2012 http://www.sciencedirect.com/science/article/pii/S0166685112000539

  2. Bats, Trypanosomes, and Triatomines in Ecuador: New Insights into the Diversity, Transmission, and Origins of Trypanosoma cruzi and Chagas Disease.

    Directory of Open Access Journals (Sweden)

    C Miguel Pinto

    Full Text Available The generalist parasite Trypanosoma cruzi has two phylogenetic lineages associated almost exclusively with bats-Trypanosoma cruzi Tcbat and the subspecies T. c. marinkellei. We present new information on the genetic variation, geographic distribution, host associations, and potential vectors of these lineages. We conducted field surveys of bats and triatomines in southern Ecuador, a country endemic for Chagas disease, and screened for trypanosomes by microscopy and PCR. We identified parasites at species and genotype levels through phylogenetic approaches based on 18S ribosomal RNA (18S rRNA and cytochrome b (cytb genes and conducted a comparison of nucleotide diversity of the cytb gene. We document for the first time T. cruzi Tcbat and T. c. marinkellei in Ecuador, expanding their distribution in South America to the western side of the Andes. In addition, we found the triatomines Cavernicola pilosa and Triatoma dispar sharing shelters with bats. The comparisons of nucleotide diversity revealed a higher diversity for T. c. marinkellei than any of the T. c. cruzi genotypes associated with Chagas disease. Findings from this study increased both the number of host species and known geographical ranges of both parasites and suggest potential vectors for these two trypanosomes associated with bats in rural areas of southern Ecuador. The higher nucleotide diversity of T. c. marinkellei supports a long evolutionary relationship between T. cruzi and bats, implying that bats are the original hosts of this important parasite.

  3. Bats, Trypanosomes, and Triatomines in Ecuador: New Insights into the Diversity, Transmission, and Origins of Trypanosoma cruzi and Chagas Disease.

    Science.gov (United States)

    Pinto, C Miguel; Ocaña-Mayorga, Sofía; Tapia, Elicio E; Lobos, Simón E; Zurita, Alejandra P; Aguirre-Villacís, Fernanda; MacDonald, Amber; Villacís, Anita G; Lima, Luciana; Teixeira, Marta M G; Grijalva, Mario J; Perkins, Susan L

    2015-01-01

    The generalist parasite Trypanosoma cruzi has two phylogenetic lineages associated almost exclusively with bats-Trypanosoma cruzi Tcbat and the subspecies T. c. marinkellei. We present new information on the genetic variation, geographic distribution, host associations, and potential vectors of these lineages. We conducted field surveys of bats and triatomines in southern Ecuador, a country endemic for Chagas disease, and screened for trypanosomes by microscopy and PCR. We identified parasites at species and genotype levels through phylogenetic approaches based on 18S ribosomal RNA (18S rRNA) and cytochrome b (cytb) genes and conducted a comparison of nucleotide diversity of the cytb gene. We document for the first time T. cruzi Tcbat and T. c. marinkellei in Ecuador, expanding their distribution in South America to the western side of the Andes. In addition, we found the triatomines Cavernicola pilosa and Triatoma dispar sharing shelters with bats. The comparisons of nucleotide diversity revealed a higher diversity for T. c. marinkellei than any of the T. c. cruzi genotypes associated with Chagas disease. Findings from this study increased both the number of host species and known geographical ranges of both parasites and suggest potential vectors for these two trypanosomes associated with bats in rural areas of southern Ecuador. The higher nucleotide diversity of T. c. marinkellei supports a long evolutionary relationship between T. cruzi and bats, implying that bats are the original hosts of this important parasite.

  4. Regulation and spatial organization of PCNA in Trypanosoma brucei

    International Nuclear Information System (INIS)

    Kaufmann, Doris; Gassen, Alwine; Maiser, Andreas; Leonhardt, Heinrich; Janzen, Christian J.

    2012-01-01

    Highlights: ► Characterization of the proliferating cell nuclear antigen in Trypanosoma brucei (TbPCNA). ► TbPCNA is a suitable marker to detect replication in T. brucei. ► TbPCNA distribution and regulation is different compared to closely related parasites T. cruzi and Leishmania donovani. -- Abstract: As in most eukaryotic cells, replication is regulated by a conserved group of proteins in the early-diverged parasite Trypanosoma brucei. Only a few components of the replication machinery have been described in this parasite and regulation, sub-nuclear localization and timing of replication are not well understood. We characterized the proliferating cell nuclear antigen in T. brucei (TbPCNA) to establish a spatial and temporal marker for replication. Interestingly, PCNA distribution and regulation is different compared to the closely related parasites Trypanosoma cruzi and Leishmania donovani. TbPCNA foci are clearly detectable during S phase of the cell cycle but in contrast to T. cruzi they are not preferentially located at the nuclear periphery. Furthermore, PCNA seems to be degraded when cells enter G2 phase in T. brucei suggesting different modes of replication regulation or functions of PCNA in these closely related eukaryotes.

  5. [Esophageal motor disorders in asymptomatic subjects with Trypanosoma cruzi infection].

    Science.gov (United States)

    Torres-Aguilera, M; Remes-Troche, J M; Roesch-Dietlen, F; Vázquez-Jiménez, J G; De la Cruz-Patiño, E; Grube-Pagola, P; Ruiz-Juárez, I

    2011-01-01

    The indeterminate chronic or "asymptomatic" phase of Trypanosoma cruzi (Chagas' disease) infection is characterized by the absence of gastrointestinal symptoms, and has an estimated duration of 20 to 30 years. However, the intramural denervation that induces dysfunction of the gastrointestinal tract is progressive. Recently, epidemiological studies have shown that the seroprevalence for this infection in our area ranges between 2% and 3% of the population. To detect the presence of esophageal motor disorders in asymptomatic individuals chronically infected with Trypanosoma cruzi using standard esophageal manometry. A cross sectional study in 28 asymptomatic subjects (27 men, age 40.39 ± 10.79) with serological evidence of infection with Trypanosoma cruzi was performed. In all cases demographic characteristics, gastrointestinal symptoms and esophageal motility disorders using conventional manometry were analyzed. In this study 54% (n = 15) of asymptomatic subjects had an esophageal motor disorder: 5 (18%) had nutcracker esophagus, 5 (18%) nonspecific esophageal motor disorders, 3 (11%) hypertensive lower esophageal sphincter (LES), 1 (4%) an incomplete relaxation of the LES and 1 (4%) had chagasic achalasia. More than half of patients that course with Chagas' disease in the indeterminate phase and that are apparently asymptomatic have impaired esophageal motility. Presence of hypertensive LES raises the possibility that this alteration represents an early stage in the development of chagasic achalasia.

  6. Regulation and spatial organization of PCNA in Trypanosoma brucei

    Energy Technology Data Exchange (ETDEWEB)

    Kaufmann, Doris; Gassen, Alwine [University of Munich (LMU), Department Biology I, Genetics, Grosshaderner Str. 2-4, 82152 Martinsried (Germany); Maiser, Andreas; Leonhardt, Heinrich [University of Munich (LMU), Department Biology II, Grosshaderner Str. 2-4, 82152 Martinsried (Germany); Janzen, Christian J., E-mail: christian.janzen@uni-wuerzburg.de [University of Munich (LMU), Department Biology I, Genetics, Grosshaderner Str. 2-4, 82152 Martinsried (Germany)

    2012-03-23

    Highlights: Black-Right-Pointing-Pointer Characterization of the proliferating cell nuclear antigen in Trypanosoma brucei (TbPCNA). Black-Right-Pointing-Pointer TbPCNA is a suitable marker to detect replication in T. brucei. Black-Right-Pointing-Pointer TbPCNA distribution and regulation is different compared to closely related parasites T. cruzi and Leishmania donovani. -- Abstract: As in most eukaryotic cells, replication is regulated by a conserved group of proteins in the early-diverged parasite Trypanosoma brucei. Only a few components of the replication machinery have been described in this parasite and regulation, sub-nuclear localization and timing of replication are not well understood. We characterized the proliferating cell nuclear antigen in T. brucei (TbPCNA) to establish a spatial and temporal marker for replication. Interestingly, PCNA distribution and regulation is different compared to the closely related parasites Trypanosoma cruzi and Leishmania donovani. TbPCNA foci are clearly detectable during S phase of the cell cycle but in contrast to T. cruzi they are not preferentially located at the nuclear periphery. Furthermore, PCNA seems to be degraded when cells enter G2 phase in T. brucei suggesting different modes of replication regulation or functions of PCNA in these closely related eukaryotes.

  7. Identification of compounds with anti-proliferative activity against Trypanosoma brucei brucei strain 427 by a whole cell viability based HTS campaign.

    Directory of Open Access Journals (Sweden)

    Melissa L Sykes

    Full Text Available Human African Trypanosomiasis (HAT is caused by two trypanosome sub-species, Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense. Drugs available for the treatment of HAT have significant issues related to difficult administration regimes and limited efficacy across species and disease stages. Hence, there is considerable need to find new alternative and less toxic drugs. An approach to identify starting points for new drug candidates is high throughput screening (HTS of large compound library collections. We describe the application of an Alamar Blue based, 384-well HTS assay to screen a library of 87,296 compounds against the related trypanosome subspecies, Trypanosoma brucei brucei bloodstream form lister 427. Primary hits identified against T.b. brucei were retested and the IC(50 value compounds were estimated for T.b. brucei and a mammalian cell line HEK293, to determine a selectivity index for each compound. The screening campaign identified 205 compounds with greater than 10 times selectivity against T.b. brucei. Cluster analysis of these compounds, taking into account chemical and structural properties required for drug-like compounds, afforded a panel of eight compounds for further biological analysis. These compounds had IC(50 values ranging from 0.22 µM to 4 µM with associated selectivity indices ranging from 19 to greater than 345. Further testing against T.b. rhodesiense led to the selection of 6 compounds from 5 new chemical classes with activity against the causative species of HAT, which can be considered potential candidates for HAT early drug discovery. Structure activity relationship (SAR mining revealed components of those hit compound structures that may be important for biological activity. Four of these compounds have undergone further testing to 1 determine whether they are cidal or static in vitro at the minimum inhibitory concentration (MIC, and 2 estimate the time to kill.

  8. Rational Design of a New Trypanosoma rangeli Trans-Sialidase for Efficient Sialylation of Glycans

    DEFF Research Database (Denmark)

    Jers, Carsten; Michalak, Malwina; Larsen, Dorte Møller

    2014-01-01

    This paper reports rational engineering of Trypanosoma rangeli sialidase to develop an effective enzyme for a potentially important type of reactivity: production of sialylated prebiotic glycans. The Trypanosoma cruzi trans-sialidase and the homologous T. rangeli sialidase has previously been use...

  9. Melophagus ovinus and Trypanosoma (Megatrypanum) melophagium in ovines in the State of Minas Gerais, Brasil

    OpenAIRE

    Costa, José Oswaldo; Lima, Walter dos Santos; Leite, Antonio César Rios; Guimarães, Marcos Pezzi; Torres, Liléia Diotaiuti

    1983-01-01

    Neste trabalho Melophagus ovinus é identificado pela primeira vez no Estado de Minas Gerais e Trypanosoma (Megatrypanum) melophagium tem sua primeira ocorrência registrada no Brasil.Melophagus ovinus is identified for the first time in Minas Gerais State and Trypanosoma (Megatrypanum) melophagium in Brazil.

  10. Depression Screening

    Science.gov (United States)

    ... Depression Screening Substance Abuse Screening Alcohol Use Screening Depression Screening (PHQ-9) - Instructions The following questions are ... this tool, there is also text-only version . Depression Screening - Manual Instructions The following questions are a ...

  11. Discovery and Optimization of 5-Amino-1,2,3-triazole-4-carboxamide Series against Trypanosoma cruzi.

    Science.gov (United States)

    Brand, Stephen; Ko, Eun Jung; Viayna, Elisabet; Thompson, Stephen; Spinks, Daniel; Thomas, Michael; Sandberg, Lars; Francisco, Amanda F; Jayawardhana, Shiromani; Smith, Victoria C; Jansen, Chimed; De Rycker, Manu; Thomas, John; MacLean, Lorna; Osuna-Cabello, Maria; Riley, Jennifer; Scullion, Paul; Stojanovski, Laste; Simeons, Frederick R C; Epemolu, Ola; Shishikura, Yoko; Crouch, Sabrinia D; Bakshi, Tania S; Nixon, Christopher J; Reid, Iain H; Hill, Alan P; Underwood, Tim Z; Hindley, Sean J; Robinson, Sharon A; Kelly, John M; Fiandor, Jose M; Wyatt, Paul G; Marco, Maria; Miles, Timothy J; Read, Kevin D; Gilbert, Ian H

    2017-09-14

    Chagas' disease, caused by the protozoan parasite Trypanosoma cruzi, is the most common cause of cardiac-related deaths in endemic regions of Latin America. There is an urgent need for new safer treatments because current standard therapeutic options, benznidazole and nifurtimox, have significant side effects and are only effective in the acute phase of the infection with limited efficacy in the chronic phase. Phenotypic high content screening against the intracellular parasite in infected VERO cells was used to identify a novel hit series of 5-amino-1,2,3-triazole-4-carboxamides (ATC). Optimization of the ATC series gave improvements in potency, aqueous solubility, and metabolic stability, which combined to give significant improvements in oral exposure. Mitigation of a potential Ames and hERG liability ultimately led to two promising compounds, one of which demonstrated significant suppression of parasite burden in a mouse model of Chagas' disease.

  12. In Silico Identification and in Vitro Activity of Novel Natural Inhibitors of Trypanosoma brucei Glyceraldehyde-3-phosphate-dehydrogenase

    Directory of Open Access Journals (Sweden)

    Fabian C. Herrmann

    2015-09-01

    Full Text Available As part of our ongoing efforts to identify natural products with activity against pathogens causing neglected tropical diseases, we are currently performing an extensive screening of natural product (NP databases against a multitude of protozoan parasite proteins. Within this project, we screened a database of NPs from a commercial supplier, AnalytiCon Discovery (Potsdam, Germany, against Trypanosoma brucei glyceraldehyde-3-phosphate dehydrogenase (TbGAPDH, a glycolytic enzyme whose inhibition deprives the parasite of energy supply. NPs acting as potential inhibitors of the mentioned enzyme were identified using a pharmacophore-based virtual screening and subsequent docking of the identified hits into the active site of interest. In a set of 700 structures chosen for the screening, 13 (1.9% were predicted to possess significant affinity towards the enzyme and were therefore tested in an in vitro enzyme assay using recombinant TbGAPDH. Nine of these in silico hits (69% showed significant inhibitory activity at 50 µM, of which two geranylated benzophenone derivatives proved to be particularly active with IC50 values below 10 µM. These compounds also showed moderate in vitro activity against T. brucei rhodesiense and may thus represent interesting starting points for further optimization.

  13. In Silico Identification and in Vitro Activity of Novel Natural Inhibitors of Trypanosoma brucei Glyceraldehyde-3-phosphate-dehydrogenase.

    Science.gov (United States)

    Herrmann, Fabian C; Lenz, Mairin; Jose, Joachim; Kaiser, Marcel; Brun, Reto; Schmidt, Thomas J

    2015-09-03

    As part of our ongoing efforts to identify natural products with activity against pathogens causing neglected tropical diseases, we are currently performing an extensive screening of natural product (NP) databases against a multitude of protozoan parasite proteins. Within this project, we screened a database of NPs from a commercial supplier, AnalytiCon Discovery (Potsdam, Germany), against Trypanosoma brucei glyceraldehyde-3-phosphate dehydrogenase (TbGAPDH), a glycolytic enzyme whose inhibition deprives the parasite of energy supply. NPs acting as potential inhibitors of the mentioned enzyme were identified using a pharmacophore-based virtual screening and subsequent docking of the identified hits into the active site of interest. In a set of 700 structures chosen for the screening, 13 (1.9%) were predicted to possess significant affinity towards the enzyme and were therefore tested in an in vitro enzyme assay using recombinant TbGAPDH. Nine of these in silico hits (69%) showed significant inhibitory activity at 50 µM, of which two geranylated benzophenone derivatives proved to be particularly active with IC50 values below 10 µM. These compounds also showed moderate in vitro activity against T. brucei rhodesiense and may thus represent interesting starting points for further optimization.

  14. Perspectives on the Trypanosoma cruzi–host cell receptor interactions

    Science.gov (United States)

    Villalta, Fernando; Scharfstein, Julio; Ashton, Anthony W.; Tyler, Kevin M.; Guan, Fangxia; Mukherjee, Shankar; Lima, Maria F.; Alvarez, Sandra; Weiss, Louis M.; Huang, Huan; Machado, Fabiana S.

    2009-01-01

    Chagas disease is caused by the parasite Trypanosoma cruzi. The critical initial event is the interaction of the trypomastigote form of the parasite with host receptors. This review highlights recent observations concerning these interactions. Some of the key receptors considered are those for thromboxane, bradykinin, and for the nerve growth factor TrKA. Other important receptors such as galectin-3, thrombospondin, and laminin are also discussed. Investigation into the molecular biology and cell biology of host receptors for T. cruzi may provide novel therapeutic targets. PMID:19283409

  15. Trypanosoma sp. diversity in Amazonian bats (Chiroptera; Mammalia) from Acre State, Brazil.

    Science.gov (United States)

    Dos Santos, Francisco C B; Lisboa, Cristiane V; Xavier, Samanta C C; Dario, Maria A; Verde, Rair de S; Calouro, Armando M; Roque, André Luiz R; Jansen, Ana M

    2017-11-16

    Bats are ancient hosts of Trypanosoma species and their flying ability, longevity and adaptability to distinct environments indicate that they are efficient dispersers of parasites. Bats from Acre state (Amazon Biome) were collected in four expeditions conducted in an urban forest (Parque Zoobotânico) and one relatively more preserved area (Seringal Cahoeira) in Rio Branco and Xapuri municipalities. Trypanosoma sp. infection was detected by hemoculture and fresh blood examination. Isolated parasite species were identified by the similarity of the obtained DNA sequence from 18S rDNA polymerase chain reaction and reference strains. Overall, 367 bats from 23 genera and 32 species were examined. Chiropterofauna composition was specific to each municipality, although Artibeus sp. and Carollia sp. prevailed throughout. Trypanosoma sp. infection was detected in 85 bats (23·2%). The most widely distributed and prevalent genotypes were (in order) Trypanosoma cruzi TcI, T. cruzi marinkellei, Trypanosoma dionisii, T. cruzi TcIV and Trypanosoma rangeli. At least one still-undescribed Trypanosoma species was also detected in this study. The detection of T. cruzi TcI and TcIV (the ones associated with Chagas disease in Amazon biome) demonstrates the putative importance of these mammal hosts in the epidemiology of the disease in the Acre State.

  16. Immobilization of NTPDase-1 from Trypanosoma cruzi and Development of an Online Label-Free Assay.

    Science.gov (United States)

    Calil, Felipe Antunes; Lima, Juliana Maria; de Oliveira, Arthur Henrique Cavalcante; Mariotini-Moura, Christiane; Fietto, Juliana Lopes Rangel; Cardoso, Carmen Lucia

    2016-01-01

    The use of IMERs (Immobilized Enzyme Reactors) as a stationary phase coupled to high performance chromatographic systems is an interesting approach in the screening of new ligands. In addition, IMERs offer many advantages over techniques that employ enzymes in solution. The enzyme nucleoside triphosphate diphosphohydrolase (NTPDase-1) from Trypanosoma cruzi acts as a pathogen infection facilitator, so it is a good target in the search for inhibitors. In this paper, immobilization of NTPDase-1 afforded ICERs (Immobilized Capillary Enzyme Reactors). A liquid chromatography method was developed and validated to monitor the ICER activity. The conditions for the application of these bioreactors were investigated, and excellent results were obtained. The enzyme was successfully immobilized, as attested by the catalytic activity detected in the Tc NTPDase-1-ICER chromatographic system. Kinetic studies on the substrate ATP gave K M of 0.317 ± 0.044 mmol·L -1 , which still presented high affinity compared to in solution. Besides that, the ICER was stable for 32 days, enough time to investigate samples of possible inhibitors, including especially the compound Suramin, that inhibited 51% the enzyme activity at 100  µ mol·L -1 , which is in accordance with the data for the enzyme in solution.

  17. Immobilization of NTPDase-1 from Trypanosoma cruzi and Development of an Online Label-Free Assay

    Directory of Open Access Journals (Sweden)

    Felipe Antunes Calil

    2016-01-01

    Full Text Available The use of IMERs (Immobilized Enzyme Reactors as a stationary phase coupled to high performance chromatographic systems is an interesting approach in the screening of new ligands. In addition, IMERs offer many advantages over techniques that employ enzymes in solution. The enzyme nucleoside triphosphate diphosphohydrolase (NTPDase-1 from Trypanosoma cruzi acts as a pathogen infection facilitator, so it is a good target in the search for inhibitors. In this paper, immobilization of NTPDase-1 afforded ICERs (Immobilized Capillary Enzyme Reactors. A liquid chromatography method was developed and validated to monitor the ICER activity. The conditions for the application of these bioreactors were investigated, and excellent results were obtained. The enzyme was successfully immobilized, as attested by the catalytic activity detected in the TcNTPDase-1-ICER chromatographic system. Kinetic studies on the substrate ATP gave KM of 0.317 ± 0.044 mmol·L−1, which still presented high affinity compared to in solution. Besides that, the ICER was stable for 32 days, enough time to investigate samples of possible inhibitors, including especially the compound Suramin, that inhibited 51% the enzyme activity at 100 µmol·L−1, which is in accordance with the data for the enzyme in solution.

  18. Identification of TOEFAZ1-interacting proteins reveals key regulators of Trypanosoma brucei cytokinesis.

    Science.gov (United States)

    Hilton, Nicholas A; Sladewski, Thomas E; Perry, Jenna A; Pataki, Zemplen; Sinclair-Davis, Amy N; Muniz, Richard S; Tran, Holly L; Wurster, Jenna I; Seo, Jiwon; de Graffenried, Christopher L

    2018-05-21

    The protist parasite Trypanosoma brucei is an obligate extracellular pathogen that retains its highly-polarized morphology during cell division and has evolved a novel cytokinetic process independent of non-muscle myosin II. The polo-like kinase homolog TbPLK is essential for transmission of cell polarity during division and for cytokinesis. We previously identified a putative TbPLK substrate named Tip of the Extending FAZ 1 (TOEFAZ1) as an essential kinetoplastid-specific component of the T. brucei cytokinetic machinery. We performed a proximity-dependent biotinylation (BioID) screen using TOEFAZ1 as a means to identify additional proteins that are involved in cytokinesis. Using quantitative proteomic methods, we identified nearly 500 TOEFAZ1-proximal proteins and characterized 59 in further detail. Among the candidates, we identified an essential putative phosphatase that regulates the expression level and localization of both TOEFAZ1 and TbPLK, a previously uncharacterized protein that is necessary for the assembly of a new cell posterior, and a microtubule plus-end directed orphan kinesin that is required for completing cleavage furrow ingression. The identification of these proteins provides new insight into T. brucei cytokinesis and establishes TOEFAZ1 as a key component of this essential and uniquely-configured process in kinetoplastids. This article is protected by copyright. All rights reserved. © 2018 John Wiley & Sons Ltd.

  19. Trypanosoma cruzi benznidazole susceptibility in vitro does not predict the therapeutic outcome of human Chagas disease

    Directory of Open Access Journals (Sweden)

    Margoth Moreno

    2010-11-01

    Full Text Available 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.

  20. Immunodiagnosis of Trypanosoma cruzi (Chagas' Disease Infection in Naturally Infected Dogs

    Directory of Open Access Journals (Sweden)

    Lauricella MA

    1998-01-01

    Full Text Available This study reports on the standardization of an enzyme-linked immunosorbent assay (ELISA for detecting specific antibodies anti-Trypanosoma cruzi in naturally infected dogs. Sera from 182 mongrel dogs of all ages residing in four rural villages in Santiago del Estero, Argentina, were collected in November 1994 and preserved in buffered neutral glycerin. All sera were tested by indirect hemagglutination test (IHAT, indirect immunofluorescence test (IFAT, and ELISA using the flagellar fraction of T. cruzi as antigen. Dog sera from an area without vectorial transmission were used to calculate ELISA specificity and cut-off value. Eighty-six percent of sera had concordant results for all tests. All sera reactive for IHAT and IFAT were also reactive for ELISA, except in one case. Sera tested by ELISA when diluted 1:200 allowed a clearer division between non-reactive and reactive sera than when 1:100 with greater agreement among serologic techniques. The specificity of ELISA was 96.2%. Among 34 adult dogs with a positive xenodiagnosis, sensitivity was 94% both for ELISA and IFAT. ELISA is the first choice for screening purposes and one of the pair of techniques recommended for diagnostic studies in dog populations

  1. Synthesis and biological evaluation of some novel 1-indanone thiazolylhydrazone derivatives as anti-Trypanosoma cruzi agents.

    Science.gov (United States)

    Caputto, María E; Ciccarelli, Alejandra; Frank, Fernanda; Moglioni, Albertina G; Moltrasio, Graciela Y; Vega, Daniel; Lombardo, Elisa; Finkielsztein, Liliana M

    2012-09-01

    A series of novel 4-arylthiazolylhydrazones (TZHs) derived from 1-indanones were synthesized in good yields (66-92%) in a simple procedure using microwave irradiation and then characterized by spectroscopy studies. The compounds were evaluated for their in vitro anti-Trypanosoma cruzi activity against the epimastigote, trypomastigote and amastigote forms of the parasite. Most TZHs displayed excellent activity, and were more potent and selective than the reference drug Benznidazole, used in the current chemotherapy. Analysis of the free sterols from parasite incubated with the compounds showed that inhibition of ergosterol biosynthesis is a possible target for the action of these new TZHs. In particular, TZH 9 emerged as a promising antichagasic compound to be evaluated in animal models. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

  2. Trypanosoma cruzi: Transporte de metabolitos esenciales obtenidos del hospedador Trypanosoma cruzi: Transport of essential metabolites acquired from the host

    Directory of Open Access Journals (Sweden)

    Claudio A. Pereira

    2008-10-01

    Full Text Available El Trypanosoma cruzi es el agente causal de la enfermedad de Chagas, endémica en Argentina y en toda América Latina. Presenta numerosas características metabólicas diferenciales respecto a sus hospedadores insectos y mamíferos. Algunas de estas diferencias fueron consecuencia de millones de años de adaptación al parasitismo en los cuales estos organismos protozoarios reemplazaron, a lo largo de su evolución, muchas rutas metabólicas de biosíntesis por sistemas de transporte de metabolitos desde el hospedador. En esta revisión se describen los avances en el conocimiento de los sistemas de transporte tanto bioquímicos como también de las moléculas involucradas en dichos procesos. Se aborda con especial énfasis los transportadores de aminoácidos y poliaminas de T. cruzi de la familia AAAP (Amino Acid/Auxin Permeases ya que parece ser exclusiva de los tripanosomátidos. Teniendo en cuenta que estas moléculas se encuentran completamente ausentes en mamíferos podrían ser consideradas como potenciales blancos contra el Trypanosoma cruzi.Trypanosoma cruzi is the etiological agent of Chagas disease, a disease endemic not only in Argentina but also in all of Latinamerica. T. cruzi presents several metabolic characteristics which are completely absent in its insect vectors and in mammalian hosts. Some of these differences were acquired after millions of years of adaptation to parasitism, during which this protozoan replaced many biosynthetic routes for transport systems. In the present review, we describe the advances in the knowledge of T. cruzi transport processes and the molecules involved. In particular, we focus on aminoacid and polyamine transporters from the AAAP family (Amino Acid/Auxin Permeases, because they seem to be exclusive transporters from trypanosomatids. Taking into account that these permeases are completely absent in mammals, they could be considered as a potential target against Trypanosoma cruzi.

  3. Benznidazole induces in vitro anaerobic metabolism in Trypanosoma cruzi epimastigotes

    Directory of Open Access Journals (Sweden)

    Marina Clare Vinaud

    2017-11-01

    Full Text Available Objective: To determine the biochemical alterations of the energetic metabolism of Trypanosoma cruzi epimastigotes in vitro exposed to different concentrations of benzinidazole. Methods: Biochemical analyses were performed at 3, 6 (log phase, 9 and 12 (stationary phase days of culture. Parasites were exposed to five concentrations of benzinidazole. Glycolysis, tricarboxilic acid cycle and fatty acids oxidation pathways were quantified through chromatography. Glucose, urea and creatinine were quantified through spectrophotometric analysis. Results: Anaerobic fermentation and fatty acids oxidation were increased in the stationary phase of the culture. Benzinidazole at high concentrations induced anaerobic metabolism in the log phase of the culture while the parasites exposed to the lower concentrations preferred the citric acid cycle as energy production pathway. Benzinidazole did not influence on the proteins catabolism. Conclusions: It is possible to conclude that there are metabolic differences between evolutive forms of Trypanosoma cruzi and the main drug used for its treatment induces the anaerobic metabolism in the parasite, possibly impairing the mitochondrial pathways.

  4. Electrocardiographic alteration among first degree relatives with serologic evidence of Trypanosoma cruzi infection: a sibship study

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    Julio C. Morini

    1994-09-01

    Full Text Available To analyze whether electrocardiographic alterations (ECGA in patients with antibodies to Trypanosoma cruzi showed a patttern of familial aggregation, a sample of 379 young adults (166 men and 213 women distributed in sibships, were assessed for the presence of anti-T.cruzi antibodies, and subjected to a complete clinical examination and a standard resting electrocardiogram (ECG. Positive T. cruzi serology was detected in 165 individuals, 48 of them showing an abnormal ECG (overall prevalence 29 por cento. One hundred and eleven seropositive individuals were distributed in 45 sibships, each of them constituted by more than one seropositive sib, with ECGA being present in 34 out of these patients. Seropositive subjects with ECGA were detected in 27 sibships. Since the index case within each sibship is counted exactly once, affected individuals selected at random as propositi were extracted to calculate the prevalence of ECGA among first degree relatives of probands. Abnormal ECGs were recorded in 7 out of 45 sibs yielding a prevalence that did not differ from estimations registered in the general population or seropositive sibs. Data from the present sample show no familial aggregation for the occurrence of ECGA in patients with T.cruzi infection.

  5. Novel sterol metabolic network of Trypanosoma brucei procyclic and bloodstream forms

    Science.gov (United States)

    Nes, Craigen R.; Singha, Ujjal K.; Liu, Jialin; Ganapathy, Kulothungan; Villalta, Fernando; Waterman, Michael R.; Lepesheva, Galina I.; Chaudhuri, Minu; Nes, W. David

    2012-01-01

    Trypanosoma brucei is the protozoan parasite that causes African trypanosomiasis, a neglected disease of people and animals. Co-metabolite analysis, labelling studies using [methyl-2H3]-methionine and substrate/product specificities of the cloned 24-SMT (sterol C24-methyltransferase) and 14-SDM (sterol C14-demethylase) from T. brucei afforded an uncommon sterol metabolic network that proceeds from lanosterol and 31-norlanosterol to ETO [ergosta-5,7,25(27)-trien-3β-ol], 24-DTO [dimethyl ergosta-5,7,25(27)-trienol] and ergosterol [ergosta-5,7,22(23)-trienol]. To assess the possible carbon sources of ergosterol biosynthesis, specifically 13C-labelled specimens of lanosterol, acetate, leucine and glucose were administered to T. brucei and the 13C distributions found were in accord with the operation of the acetate–mevalonate pathway, with leucine as an alternative precursor, to ergostenols in either the insect or bloodstream form. In searching for metabolic signatures of procyclic cells, we observed that the 13C-labelling treatments induce fluctuations between the acetyl-CoA (mitochondrial) and sterol (cytosolic) synthetic pathways detected by the progressive increase in 13C-ergosterol production (control sterol synthesis that is further fluctuated in the cytosol, yielding distinct sterol profiles in relation to cell demands on growth. PMID:22176028

  6. Trypanosoma cruzi: Correlations of Biological Aspects of the Life Cycle in Mice and Triatomines

    Directory of Open Access Journals (Sweden)

    Lima Valdirene S

    1999-01-01

    Full Text Available The infection pattern in Swiss mice and Triatomine bugs (Rhodnius neglectus of eleven clones and the original stock of a Trypanosoma cruzi isolate, derived from a naturally infected Didelphis marsupialis, were biochemically and biologically characterized. The clones and the original isolate were in the same zymodeme (Z1 except that two clones were found to be in zymodeme 2 when tested with G6PDH. Although infective, neither the original isolate nor the clones were highly virulent for the mice and lesions were only observed in mice infected with the original stock and one of the clones (F8. All clones and the original isolate infected bugs well while only the original isolate and clones E2 and F3 yielded high metacyclogenesis rates. An observed correlation between absence of lesions in the mammal host and high metacyclogenesis rates in the invertebrate host suggest a evolutionary trade off i.e. a fitness increase in one trait which is accompanied by a fitness reduction in a different one. Our results suggest that in a species as heterogeneous as T. cruzi, a cooperation effect among the subpopulations should be considered.

  7. Trypanocidal Effect of Isotretinoin through the Inhibition of Polyamine and Amino Acid Transporters in Trypanosoma cruzi.

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    Chantal Reigada

    2017-03-01

    Full Text Available Polyamines are essential compounds to all living organisms and in the specific case of Trypanosoma cruzi, the causative agent of Chagas disease, they are exclusively obtained through transport processes since this parasite is auxotrophic for polyamines. Previous works reported that retinol acetate inhibits Leishmania growth and decreases its intracellular polyamine concentration. The present work describes a combined strategy of drug repositioning by virtual screening followed by in vitro assays to find drugs able to inhibit TcPAT12, the only polyamine transporter described in T. cruzi. After a screening of 3000 FDA-approved drugs, 7 retinoids with medical use were retrieved and used for molecular docking assays with TcPAT12. From the docked molecules, isotretinoin, a well-known drug used for acne treatment, showed the best interaction score with TcPAT12 and was selected for further in vitro studies. Isotretinoin inhibited the polyamine transport, as well as other amino acid transporters from the same protein family (TcAAAP, with calculated IC50 values in the range of 4.6-10.3 μM. It also showed a strong inhibition of trypomastigote burst from infected cells, with calculated IC50 of 130 nM (SI = 920 being significantly less effective on the epimastigote stage (IC50 = 30.6 μM. The effect of isotretinoin on the parasites plasma membrane permeability and on mammalian cell viability was tested, and no change was observed. Autophagosomes and apoptotic bodies were detected as part of the mechanisms of isotretinoin-induced death indicating that the inhibition of transporters by isotretinoin causes nutrient starvation that triggers autophagic and apoptotic processes. In conclusion, isotretinoin is a promising trypanocidal drug since it is a multi-target inhibitor of essential metabolites transporters, in addition to being an FDA-approved drug largely used in humans, which could reduce significantly the requirements for its possible application in the

  8. In or out? On the tightness of glycosomal compartmentalization of metabolites and enzymes in Trypanosoma brucei

    NARCIS (Netherlands)

    Haanstra, Jurgen R.; Bakker, Barbara M.; Michels, Paul A. M.

    Trypanosomatids sequester large parts of glucose metabolism inside specialised peroxisomes, called glycosomes. Many studies have shown that correct glycosomal compartmentalization of glycolytic enzymes is essential for bloodstream-form Trypanosoma brucel. The recent finding of pore-forming

  9. Novel molecular mechanism for targeting the parasite Trypanosoma brucei with snake venom toxins

    DEFF Research Database (Denmark)

    Martos Esteban, Andrea; Laustsen, Andreas Hougaard; Carrington, Mark

    Trypanosoma brucei is a parasitic protozoan species capable to infecting insect vectors whose bite further produces African sleeping sickness inhuman beings. During parasites’extracellular lives in the mammalian host, its outer coat, mainly composedof Variable surface glycoproteins (VSGs)[2...

  10. Effects of medicinal plant extracts on growth of Leishmania (L. amazonensis and Trypanosoma cruzi Efeito de extratos de plantas medicinais no crescimento de Leishmania (L. amazonensis e Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Patrícia Shima Luize

    2005-03-01

    Full Text Available This study describes the screening of extracts obtained from 19 species of plants used in Brazilian traditional medicine for treatment of a variety of diseases. The extracts were tested against axenic amastigote and promastigote forms of Leishmania (L. amazonensis, and epimastigote forms of Trypanosoma cruzi in vitro at a concentration of 100 mg/ml. Baccharis trimera, Cymbopogon citratus, Matricaria chamomilla, Mikania glomerata, Ocimum gratissimum, Piper regnellii, Prunus domestica, Psidium guajava, Sambucus canadensis, Stryphnodendron adstringens, Tanacetum parthenium, and Tanacetum vulgare showed significant effects against one or both parasites, with a percentage of growth inhibition between 49.5 and 99%. The extracts showed no cytotoxic effect on sheep erythrocytes. These medicinal plants may be sources of new compounds that are clinically active against L. amazonensis and T. cruzi.Este estudo descreve a triagem de extratos obtidos de 19 espécies de plantas usadas na medicina tradicional brasileira para o tratamento de várias doenças. Os extratos foram testados contra formas amastigota axênica e promastigota de Leishmania (L. amazonensis, e formas epimastigota de Trypanosoma cruzi in vitro na concentração de 100 mg/ml. Baccharis trimera, Cymbopogon citratus, Matricaria chamomilla, Mikania glomerata, Ocimum gratissimum, Piper regnellii, Prunus domestica, Psidium guajava, Sambucus canadensis, Stryphnodendron adstringens, Tanacetum parthenium, e Tanacetum vulgare apresentaram efeito significante contra um ou ambos parasitas, com a porcentagem de inibição de crescimento entre 49,5 e 99%. Os extratos não mostraram efeito citotóxico em hemácias de carneiro. Essas plantas medicinais podem ser fontes alternativas de novos compostos clinicamente ativos contra L. amazonensis e T. cruzi.

  11. Trypanocidal drugs for chronic asymptomatic Trypanosoma cruzi infection.

    Science.gov (United States)

    Villar, Juan Carlos; Perez, Juan Guillermo; Cortes, Olga Lucia; Riarte, Adelina; Pepper, Micah; Marin-Neto, Jose Antonio; Guyatt, Gordon H

    2014-05-27

    Prevention of chronic chagasic cardiomyopathy (CCC) by treating infected populations with trypanocidal therapy (TT) remains a challenge. Despite a renewed enthusiasm for TT, uncertainty regarding its efficacy, concerns about its safety and limited availability remain barriers for a wider use of conventional drugs. We have updated a previous version of this review. To systematically search, appraise, identify and extract data from eligible studies comparing the outcome of cohorts of seropositive individuals to Trypanosoma cruzi exposed to TT versus placebo or no treatment. We sought eligible studies in electronic databases (Cochrane Central Register of Controlled Trials (CENTRAL), Issue 1, 2014); MEDLINE (Ovid, 1946 to January week 5 2014); EMBASE (Ovid, 1980 to 2014 week 6) and LILACS (up to 6 May 2010)) by combining terms related with the disease and the treatment. The search also included a Google search, handsearch for references in review or selected articles, and search of expert files. We applied no language restrictions. Review authors screened the retrieved references for eligibility (those dealing with human participants treated with TT) and then assessed the pre-selected studies in full for inclusion. We included randomised controlled trials (RCTs) and observational studies that provided data on either mortality or clinical progression of CCC after at least four years of follow-up. Teams of two review authors independently carried out the study selection, data extraction and risk of bias assessment, with a referee resolving disagreement within the pairs. Data collection included study design, characteristics of the population and interventions or exposures and outcome measures. We defined categories of outcome data as parasite-related (positive serology, xenodiagnosis or polymerase chain reaction (PCR) after TT) and participant-related (including efficacy outcomes such as progression towards CCC, all-cause mortality and side effects of TT). We reported

  12. Mosaic VSGs and the scale of Trypanosoma brucei antigenic variation.

    Directory of Open Access Journals (Sweden)

    James P J Hall

    Full Text Available A main determinant of prolonged Trypanosoma brucei infection and transmission and success of the parasite is the interplay between host acquired immunity and antigenic variation of the parasite variant surface glycoprotein (VSG coat. About 0.1% of trypanosome divisions produce a switch to a different VSG through differential expression of an archive of hundreds of silent VSG genes and pseudogenes, but the patterns and extent of the trypanosome diversity phenotype, particularly in chronic infection, are unclear. We applied longitudinal VSG cDNA sequencing to estimate variant richness and test whether pseudogenes contribute to antigenic variation. We show that individual growth peaks can contain at least 15 distinct variants, are estimated computationally to comprise many more, and that antigenically distinct 'mosaic' VSGs arise from segmental gene conversion between donor VSG genes or pseudogenes. The potential for trypanosome antigenic variation is probably much greater than VSG archive size; mosaic VSGs are core to antigenic variation and chronic infection.

  13. Parasite Genome Projects and the Trypanosoma cruzi Genome Initiative

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    Wim Degrave

    1997-11-01

    Full Text Available Since the start of the human genome project, a great number of genome projects on other "model" organism have been initiated, some of them already completed. Several initiatives have also been started on parasite genomes, mainly through support from WHO/TDR, involving North-South and South-South collaborations, and great hopes are vested in that these initiatives will lead to new tools for disease control and prevention, as well as to the establishment of genomic research technology in developing countries. The Trypanosoma cruzi genome project, using the clone CL-Brener as starting point, has made considerable progress through the concerted action of more than 20 laboratories, most of them in the South. A brief overview of the current state of the project is given

  14. Electron Microscopy Analysis of the Nucleolus of Trypanosoma cruzi

    Science.gov (United States)

    López-Velázquez, Gabriel; Hernández, Roberto; López-Villaseñor, Imelda; Reyes-Vivas, Horacio; Segura-Valdez, María De L.; Jiménez-García, Luis F.

    2005-08-01

    The nucleolus is the main site for synthesis and processing of ribosomal RNA in eukaryotes. In mammals, plants, and yeast the nucleolus has been extensively characterized by electron microscopy, but in the majority of the unicellular eukaryotes no such studies have been performed. Here we used ultrastructural cytochemical and immunocytochemical techniques as well as three-dimensional reconstruction to analyze the nucleolus of Trypanosoma cruzi, which is an early divergent eukaryote of medical importance. In T. cruzi epimastigotes the nucleolus is a spherical intranuclear ribonucleoprotein organelle localized in a relatively central position within the nucleus. Dense fibrillar and granular components but not fibrillar centers were observed. In addition, nuclear bodies resembling Cajal bodies were observed associated to the nucleolus in the surrounding nucleoplasm. Our results provide additional morphological data to better understand the synthesis and processing of the ribosomal RNA in kinetoplastids.

  15. Lysophosphatidylcholine: A Novel Modulator of Trypanosoma cruzi Transmission

    Science.gov (United States)

    Silva-Neto, Mário A. C.; Carneiro, Alan B.; Silva-Cardoso, Livia; Atella, Georgia C.

    2012-01-01

    Lysophosphatidylcholine is a bioactive lipid that regulates a large number of cellular processes and is especially present during the deposition and infiltration of inflammatory cells and deposition of atheromatous plaque. Such molecule is also present in saliva and feces of the hematophagous organism Rhodnius prolixus, a triatominae bug vector of Chagas disease. We have recently demonstrated that LPC is a modulator of Trypanosoma cruzi transmission. It acts as a powerful chemoattractant for inflammatory cells at the site of the insect bite, which will provide a concentrated population of cells available for parasite infection. Also, LPC increases macrophage intracellular calcium concentrations that ultimately enhance parasite invasion. Finally, LPC inhibits NO production by macrophages stimulated by live T. cruzi, and thus interferes with the immune system of the vertebrate host. In the present paper, we discuss the main signaling mechanisms that are likely used by such molecule and their eventual use as targets to block parasite transmission and the pathogenesis of Chagas disease. PMID:22132309

  16. First report of Trypanosoma vegrandis in koalas (Phascolarctos cinereus).

    Science.gov (United States)

    Barbosa, Amanda; Austen, Jill; Gillett, Amber; Warren, Kristin; Paparini, Andrea; Irwin, Peter; Ryan, Una

    2016-08-01

    The present study describes the first report of Trypanosoma vegrandis in koalas using morphology and sequence analysis of the 18S rRNA gene. The prevalence of T. vegrandis in koalas was 13.6% (6/44). It is likely that the small size of T. vegrandis (<10μm in length), coupled with the difficulties in amplifying DNA of this parasite in mixed infections using trypanosome generic primers, are the reason why this organism has not been identified in koalas until now. This study highlights the importance of further research comprising a larger sample size to determine the prevalence of T. vegrandis in koalas as well as its potential impacts upon this marsupial species' health. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  17. Multiple evolutionary origins of Trypanosoma evansi in Kenya.

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    Christine M Kamidi

    2017-09-01

    Full Text Available Trypanosoma evansi is the parasite causing surra, a form of trypanosomiasis in camels and other livestock, and a serious economic burden in Kenya and many other parts of the world. Trypanosoma evansi transmission can be sustained mechanically by tabanid and Stomoxys biting flies, whereas the closely related African trypanosomes T. brucei brucei and T. b. rhodesiense require cyclical development in tsetse flies (genus Glossina for transmission. In this study, we investigated the evolutionary origins of T. evansi. We used 15 polymorphic microsatellites to quantify levels and patterns of genetic diversity among 41 T. evansi isolates and 66 isolates of T. b. brucei (n = 51 and T. b. rhodesiense (n = 15, including many from Kenya, a region where T. evansi may have evolved from T. brucei. We found that T. evansi strains belong to at least two distinct T. brucei genetic units and contain genetic diversity that is similar to that in T. brucei strains. Results indicated that the 41 T. evansi isolates originated from multiple T. brucei strains from different genetic backgrounds, implying independent origins of T. evansi from T. brucei strains. This surprising finding further suggested that the acquisition of the ability of T. evansi to be transmitted mechanically, and thus the ability to escape the obligate link with the African tsetse fly vector, has occurred repeatedly. These findings, if confirmed, have epidemiological implications, as T. brucei strains from different genetic backgrounds can become either causative agents of a dangerous, cosmopolitan livestock disease or of a lethal human disease, like for T. b. rhodesiense.

  18. Semisolid liver infusion tryptose supplemented with human urine allows growth and isolation of Trypanosoma cruzi and Trypanosoma rangeli clonal lineages

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    Emanuella Francisco Fajardo

    2016-06-01

    Full Text Available Abstract: INTRODUCTION This work shows that 3% (v/v human urine (HU in semisolid Liver Infusion Tryptose (SSL medium favors the growth of Trypanosoma cruzi and T. rangeli. METHODS Parasites were plated as individual or mixed strains on SSL medium and on SSL medium with 3% human urine (SSL-HU. Isolate DNA was analyzed using polymerase chain reaction (PCR and pulsed-field gel electrophoresis (PFGE. RESULTS SSL-HU medium improved clone isolation. PCR revealed that T. cruzi strains predominate on mixed-strain plates. PFGE confirmed that isolated parasites share the same molecular karyotype as parental cell lines. CONCLUSIONS SSL-HU medium constitutes a novel tool for obtaining T. cruzi and T. rangeli clonal lineages.

  19. Improvement of trans-sialylation versus hydrolysis activity of an engineered sialidase from Trypanosoma rangeli by use of co-solvents

    DEFF Research Database (Denmark)

    Zeuner, Birgitte; Riisager, Anders; Mikkelsen, Jørn Dalgaard

    2014-01-01

    hexafluorophosphate), were examined as co-solvents for the improvement of the synthesis versus hydrolysis ratio in the trans-sialylation of lactose, catalysed by an engineered sialidase from Trypanosoma rangeli. The use of 25 % (v/v) t-butanol as co-solvent significantly increased 3'-sialyllactose production by 40...... % from 1.04 ± 0.09 to 1.47 ± 0.01 mM. The synthesis versus hydrolysis ratio increased correspondingly by 1.2-times. 1-2.5 % (v/v) EAN or [C2OHMIm][PF6] improved the synthesis versus hydrolysis ratio up to 2.5-times but simultaneously decreased the 3'-sialyllactose yield, probably due to enzyme...... inactivation caused by the ionic liquid. [MMIm][MeSO4] had a detrimental effect on the trans-sialylation yield and on the ratio between synthesis and hydrolysis....

  20. Trypanosoma cruzi, cancer and the Cold War Trypanosoma cruzi, câncer e a Guerra Fria

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    Nikolai Krementsov

    2009-07-01

    Full Text Available In the summer of 1946, the international community of cancer researchers was inspired by the announcement that two Soviet scientists, Nina Kliueva and Grigorii Roskin, had discovered anticancer properties in culture extracts made from the South American protozoan, Trypanosoma cruzi, and had produced a preparation - named after its discoverers KR - which showed clear therapeutic effects on cancer patients. Research teams from various countries enthusiastically pursued the promising new line of investigation. The story of the rise and fall of interest in the anticancer properties of T. cruzi in different countries suggests that during the second half of the twentieth century, the Cold War competition between the superpowers played an important role in shaping the research agendas of cancer studies.No verão de 1946, a comunidade internacional que desenvolve pesquisas sobre o câncer, inspirou-se no anúncio de que dois cientistas soviéticos, Nina Kliueva e Grigorii Roskin, descobriram propriedades anticancerígenas em cultura extraída do protozoário existente na América Latina, o Trypanosoma cruzi e produziram um preparado que foi denominado com as iniciais KR - em sua homenagem. Grupos de pesquisadores de diversos países buscaram com entusiasmo as promessas dessa nova linha de investigação. A história da ascensão e queda do interesse nas propriedades anticâncer do T. cruzzi em diferentes países sugere que durante a segunda metade do século 20, a Guerra Fria teve um papel importante na definição das agendas de pesquisas sobre o câncer.

  1. A Trypanosoma brucei kinesin heavy chain promotes parasite growth by triggering host arginase activity.

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    Géraldine De Muylder

    2013-10-01

    Full Text Available In order to promote infection, the blood-borne parasite Trypanosoma brucei releases factors that upregulate arginase expression and activity in myeloid cells.By screening a cDNA library of T. brucei with an antibody neutralizing the arginase-inducing activity of parasite released factors, we identified a Kinesin Heavy Chain isoform, termed TbKHC1, as responsible for this effect. Following interaction with mouse myeloid cells, natural or recombinant TbKHC1 triggered SIGN-R1 receptor-dependent induction of IL-10 production, resulting in arginase-1 activation concomitant with reduction of nitric oxide (NO synthase activity. This TbKHC1 activity was IL-4Rα-independent and did not mirror M2 activation of myeloid cells. As compared to wild-type T. brucei, infection by TbKHC1 KO parasites was characterized by strongly reduced parasitaemia and prolonged host survival time. By treating infected mice with ornithine or with NO synthase inhibitor, we observed that during the first wave of parasitaemia the parasite growth-promoting effect of TbKHC1-mediated arginase activation resulted more from increased polyamine production than from reduction of NO synthesis. In late stage infection, TbKHC1-mediated reduction of NO synthesis appeared to contribute to liver damage linked to shortening of host survival time.A kinesin heavy chain released by T. brucei induces IL-10 and arginase-1 through SIGN-R1 signaling in myeloid cells, which promotes early trypanosome growth and favors parasite settlement in the host. Moreover, in the late stage of infection, the inhibition of NO synthesis by TbKHC1 contributes to liver pathogenicity.

  2. An Atypical Mitochondrial Carrier That Mediates Drug Action in Trypanosoma brucei.

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    Juan P de Macêdo

    2015-05-01

    Full Text Available Elucidating the mechanism of action of trypanocidal compounds is an important step in the development of more efficient drugs against Trypanosoma brucei. In a screening approach using an RNAi library in T. brucei bloodstream forms, we identified a member of the mitochondrial carrier family, TbMCP14, as a prime candidate mediating the action of a group of anti-parasitic choline analogs. Depletion of TbMCP14 by inducible RNAi in both bloodstream and procyclic forms increased resistance of parasites towards the compounds by 7-fold and 3-fold, respectively, compared to uninduced cells. In addition, down-regulation of TbMCP14 protected bloodstream form mitochondria from a drug-induced decrease in mitochondrial membrane potential. Conversely, over-expression of the carrier in procyclic forms increased parasite susceptibility more than 13-fold. Metabolomic analyses of parasites over-expressing TbMCP14 showed increased levels of the proline metabolite, pyrroline-5-carboxylate, suggesting a possible involvement of TbMCP14 in energy production. The generation of TbMCP14 knock-out parasites showed that the carrier is not essential for survival of T. brucei bloodstream forms, but reduced parasite proliferation under standard culture conditions. In contrast, depletion of TbMCP14 in procyclic forms resulted in growth arrest, followed by parasite death. The time point at which parasite proliferation stopped was dependent on the major energy source, i.e. glucose versus proline, in the culture medium. Together with our findings that proline-dependent ATP production in crude mitochondria from TbMCP14-depleted trypanosomes was reduced compared to control mitochondria, the study demonstrates that TbMCP14 is involved in energy production in T. brucei. Since TbMCP14 belongs to a trypanosomatid-specific clade of mitochondrial carrier family proteins showing very poor similarity to mitochondrial carriers of mammals, it may represent an interesting target for drug

  3. RAS - Screens & Assays - Drug Discovery

    Science.gov (United States)

    The RAS Drug Discovery group aims to develop assays that will reveal aspects of RAS biology upon which cancer cells depend. Successful assay formats are made available for high-throughput screening programs to yield potentially effective drug compounds.

  4. Melophagus ovinus e Trypanosoma (Megatrypanum melophagium em ovinos no Estado de Minas Gerais, Brasil Melophagus ovinus and Trypanosoma (Megatrypanum melophagium in ovines in the State of Minas Gerais, Brasil

    Directory of Open Access Journals (Sweden)

    José Oswaldo Costa

    1983-03-01

    Full Text Available Neste trabalho Melophagus ovinus é identificado pela primeira vez no Estado de Minas Gerais e Trypanosoma (Megatrypanum melophagium tem sua primeira ocorrência registrada no Brasil.Melophagus ovinus is identified for the first time in Minas Gerais State and Trypanosoma (Megatrypanum melophagium in Brazil.

  5. Engineering aspects of Passavant screening

    International Nuclear Information System (INIS)

    Siddle, K.R.; Sharma, R.K.

    1978-01-01

    The Passavant screen was developed in Europe almost 30 years ago. The Passavant screen is a vertical traveling screen; however, the basic difference between the conventional vertical traveling screen and the Passavant screen is that in the conventional screen water passes through the front screen belt and then the back screen belt, whereas in the Passavant screen the water enters in between the two belts and passes laterally through either of the belts. Thus, theoretically, the screening surface of the Passavant screen is doubled as compared to the same size conventional vertical traveling screen. Various design and operational modifications of the Passavant screen are possible to yield optimum design and performance characteristics which make it amenable to installation at power plants for safe removal of not only fish but also smaller organisms such as fish eggs and larvae. In this paper, details of the screen design and operational characteristics are discussed with notes on how these features can be modified to suit site- and organism-specific requirements

  6. Structural basis for the high specificity of a Trypanosoma congolense immunoassay targeting glycosomal aldolase.

    Directory of Open Access Journals (Sweden)

    Joar Pinto

    2017-09-01

    Full Text Available Animal African trypanosomosis (AAT is a neglected tropical disease which imposes a heavy burden on the livestock industry in Sub-Saharan Africa. Its causative agents are Trypanosoma parasites, with T. congolense and T. vivax being responsible for the majority of the cases. Recently, we identified a Nanobody (Nb474 that was employed to develop a homologous sandwich ELISA targeting T. congolense fructose-1,6-bisphosphate aldolase (TcoALD. Despite the high sequence identity between trypanosomatid aldolases, the Nb474-based immunoassay is highly specific for T. congolense detection. The results presented in this paper yield insights into the molecular principles underlying the assay's high specificity.The structure of the Nb474-TcoALD complex was determined via X-ray crystallography. Together with analytical gel filtration, the structure reveals that a single TcoALD tetramer contains four binding sites for Nb474. Through a comparison with the crystal structures of two other trypanosomatid aldolases, TcoALD residues Ala77 and Leu106 were identified as hot spots for specificity. Via ELISA and surface plasmon resonance (SPR, we demonstrate that mutation of these residues does not abolish TcoALD recognition by Nb474, but does lead to a lack of detection in the Nb474-based homologous sandwich immunoassay.The results show that the high specificity of the Nb474-based immunoassay is not determined by the initial recognition event between Nb474 and TcoALD, but rather by its homologous sandwich design. This (i provides insights into the optimal set-up of the assay, (ii may be of great significance for field applications as it could explain the potential detection escape of certain T. congolense strains, and (iii may be of general interest to those developing similar assays.

  7. Mitochondrial membrane potential-based genome-wide RNAi screen of Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Verner, Zdeněk; Paris, Zdeněk; Lukeš, Julius

    2010-01-01

    Roč. 106, č. 5 (2010), s. 1241-1244 ISSN 0932-0113 Institutional research plan: CEZ:AV0Z60220518 Keywords : GENE-FUNCTION * INTERFERENCE * mitochondrion * SUBUNITS Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.812, year: 2010

  8. Kinetoplast adaptations in American strains from Trypanosoma vivax

    Energy Technology Data Exchange (ETDEWEB)

    Greif, Gonzalo [Unidad de Biología Molecular, Institut Pasteur de Montevideo (Uruguay); Rodriguez, Matías [Sección Biomatemática, Facultad de Ciencias, Universidad de la Republica (Uruguay); Reyna-Bello, Armando [Departamento de Ciencias de la Vida, Carrera en Ingeniería en Biotecnología, Universidad de las Fuerzas Armadas (Ecuador); Centro de Estudios Biomédicos y Veterinarios, Universidad Nacional Experimental Simón Rodríguez-IDECYT, Caracas (Venezuela, Bolivarian Republic of); Robello, Carlos [Unidad de Biología Molecular, Institut Pasteur de Montevideo (Uruguay); Departamento de Bioquímica, Facultad de Medicina, Universidad de la República Uruguay (Uruguay); Alvarez-Valin, Fernando, E-mail: falvarez@fcien.edu.uy [Sección Biomatemática, Facultad de Ciencias, Universidad de la Republica (Uruguay)

    2015-03-15

    Highlights: • American T. vivax strains exhibit a drastic process of mitochondrial genome degradation. • T. vivax mitochondrial genes have among the fastest evolutionary rates in eukaryotes. • High rates of kDNA evolution are associated with relaxation of selective constrains. • Relaxed selective pressures are the result of mechanical transmission. • The evolutionary strategy of T. vivax differs from that of T. brucei-species complex. - Abstract: The mitochondrion role changes during the digenetic life cycle of African trypanosomes. Owing to the low abundance of glucose in the insect vector (tsetse flies) the parasites are dependent upon a fully functional mitochondrion, capable of performing oxidative phosphorylation. Nevertheless, inside the mammalian host (bloodstream forms), which is rich in nutrients, parasite proliferation relies on glycolysis, and the mitochondrion is partially redundant. In this work we perform a comparative study of the mitochondrial genome (kinetoplast) in different strains of Trypanosoma vivax. The comparison was conducted between a West African strain that goes through a complete life cycle and two American strains that are mechanically transmitted (by different vectors) and remain as bloodstream forms only. It was found that while the African strain has a complete and apparently fully functional kinetoplast, the American T. vivax strains have undergone a drastic process of mitochondrial genome degradation, in spite of the recent introduction of these parasites in America. Many of their genes exhibit different types of mutations that are disruptive of function such as major deletions, frameshift causing indels and missense mutations. Moreover, all but three genes (A6-ATPase, RPS12 and MURF2) are not edited in the American strains, whereas editing takes place normally in all (editable) genes from the African strain. Two of these genes, A6-ATPase and RPS12, are known to play an essential function during bloodstream stage

  9. Kinetoplast adaptations in American strains from Trypanosoma vivax

    International Nuclear Information System (INIS)

    Greif, Gonzalo; Rodriguez, Matías; Reyna-Bello, Armando; Robello, Carlos; Alvarez-Valin, Fernando

    2015-01-01

    Highlights: • American T. vivax strains exhibit a drastic process of mitochondrial genome degradation. • T. vivax mitochondrial genes have among the fastest evolutionary rates in eukaryotes. • High rates of kDNA evolution are associated with relaxation of selective constrains. • Relaxed selective pressures are the result of mechanical transmission. • The evolutionary strategy of T. vivax differs from that of T. brucei-species complex. - Abstract: The mitochondrion role changes during the digenetic life cycle of African trypanosomes. Owing to the low abundance of glucose in the insect vector (tsetse flies) the parasites are dependent upon a fully functional mitochondrion, capable of performing oxidative phosphorylation. Nevertheless, inside the mammalian host (bloodstream forms), which is rich in nutrients, parasite proliferation relies on glycolysis, and the mitochondrion is partially redundant. In this work we perform a comparative study of the mitochondrial genome (kinetoplast) in different strains of Trypanosoma vivax. The comparison was conducted between a West African strain that goes through a complete life cycle and two American strains that are mechanically transmitted (by different vectors) and remain as bloodstream forms only. It was found that while the African strain has a complete and apparently fully functional kinetoplast, the American T. vivax strains have undergone a drastic process of mitochondrial genome degradation, in spite of the recent introduction of these parasites in America. Many of their genes exhibit different types of mutations that are disruptive of function such as major deletions, frameshift causing indels and missense mutations. Moreover, all but three genes (A6-ATPase, RPS12 and MURF2) are not edited in the American strains, whereas editing takes place normally in all (editable) genes from the African strain. Two of these genes, A6-ATPase and RPS12, are known to play an essential function during bloodstream stage

  10. 6 Grain Yield

    African Journals Online (AJOL)

    create a favourable environment for rice ... developing lines adaptable to many ... have stable, not too short crop duration with ..... Analysis of variance of the effect of site and season on maturity, grain yield and plant ..... and yield components.

  11. Antitrypanosomal activity of Verbascum sinaiticum Benth. (Scrophulariaceae) against Trypanosoma congolense isolates.

    Science.gov (United States)

    Mergia, Ermias; Shibeshi, Workineh; Terefe, Getachew; Teklehaymanot, Tilahun

    2016-09-15

    African Trypanosomiasis is a neglected tropical disease with a large impact on the livelihood of the rural poor in Sub-Saharan Africa. The available drugs for managing this disease are old, expensive and are facing the problem of drug resistance. Thus, the aim of this study was to evaluate in vivo antitrypanosomal efficacy of aqueous and absolute methanol leaf extracts of Verbascum sinaiticum Benth. against Trypanosoma congolense field isolate. Verbascum sinaiticum (Local name 'qetetina') is a biennial plant, and 60-150 cm tall. It is traditionally used to treat wound, stomachache, viral infection, cancer, sunstroke, fever, abdominal colic, diarrhea, hemorrhage, anthrax, and hepatitis. The efficacy of aqueous and absolute methanol leaf extracts of V. sinaiticum was evaluated in a randomized experiment with Swiss albino mice infected with T. congolense field isolate. The extracts were administered at doses of 100, 200 and 400 mg/kg by intraperitoneal injection for seven days at 12 Days Post-Infection (DPI) when the peak parasitaemia level was approximately 10(8) trypanosomes/ml. Parasitaemia, Packed Cell Volume (PCV), mean survival time and change in body weight were used as indices for monitoring the efficacy of the extracts. Diminazene (28 mg/kg) was used as a positive control while 2 % Tween was used as the negative control. Phytochemicals screening were conducted following standard methods. The extracts showed no toxicity effect in Swiss albino mice and had LD50 above 2000 mg/kg. The phytochemicals screened in V. sinaiticum were alkaloids, flavonoids, glycoside, saponins, steroids, phenolic compounds, and tannins. The mice treated with absolute methanol leaf extract of V. sinaiticum at 400 mg/kg dose had significantly lower mean parasitaemia (7.20 ± 0.16) (p < 0.001) as compared to the negative control group (8.82 ± 0.12) on day 14 of treatment. Animals treated with the same dose had significant (p < 0.001) higher PCV value and body

  12. Natural infection of the sand fly Phlebotomus kazeruni by Trypanosoma species in Pakistan

    Directory of Open Access Journals (Sweden)

    Iwata Hiroyuki

    2010-02-01

    Full Text Available Abstract The natural infection of phlebotomine sand flies by Leishmania parasites was surveyed in a desert area of Pakistan where cutaneous leishmaniasis is endemic. Out of 220 female sand flies dissected, one sand fly, Phlebotomus kazeruni, was positive for flagellates in the hindgut. Analyses of cytochrome b (cyt b, glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH and small subunit ribosomal RNA (SSU rRNA gene sequences identified the parasite as a Trypanosoma species of probably a reptile or amphibian. This is the first report of phlebotomine sand flies naturally infected with a Trypanosoma species in Pakistan. The possible infection of sand flies with Trypanosoma species should be taken into consideration in epidemiological studies of vector species in areas where leishmaniasis is endemic.

  13. Polyclonal antibodies for the detection of Trypanosoma cruzi circulating antigens.

    Directory of Open Access Journals (Sweden)

    Edith S Málaga-Machaca

    2017-11-01

    Full Text Available Detection of Trypanosoma cruzi antigens in clinical samples is considered an important diagnostic tool for Chagas disease. The production and use of polyclonal antibodies may contribute to an increase in the sensitivity of immunodiagnosis of Chagas disease.Polyclonal antibodies were raised in alpacas, rabbits, and hens immunized with trypomastigote excreted-secreted antigen, membrane proteins, trypomastigote lysate antigen and recombinant 1F8 to produce polyclonal antibodies. Western blot analysis was performed to determine specificity of the developed antibodies. An antigen capture ELISA of circulating antigens in serum, plasma and urine samples was developed using IgY polyclonal antibodies against T. cruzi membrane antigens (capture antibody and IgG from alpaca raised against TESA. A total of 33 serum, 23 plasma and 9 urine samples were analyzed using the developed test. Among serum samples, compared to serology, the antigen capture ELISA tested positive in 55% of samples. All plasma samples from serology positive subjects were positive in the antigen capture ELISA. All urine positive samples had corresponding plasma samples that were also positive when tested by the antigen capture ELISA.Polyclonal antibodies are useful for detection of circulating antigens in both the plasma and urine of infected individuals. Detection of antigens is direct evidence of the presence of the parasite, and could be a better surrogate of current infection status.

  14. Trypanosoma avium of raptors (Falconiformes): phylogeny and identification of vectors.

    Science.gov (United States)

    Votýpka, J; Oborník, M; Volf, P; Svobodová, M; Lukes, J

    2002-09-01

    Avian trypanosomes are widespread parasites of birds, the transmission of which remains mostly unclear, with various blood-sucking insects mentioned as possible vectors. A search for vectors of trypanosomes of sparrowhawk (Accipiter nisus), buzzard (Buteo buteo), lesser-spotted eagle (Aquila pomarina) and kestrel (Falco tinnunculus) was performed in Czech and Slovak Republics. Black flies (Eusimulium spp.), hippoboscid flies (Ornithomyia avicularia), mosquitoes (Culex pipiens pipiens) and biting midges (Culicoides spp.), trapped while attempting to feed on raptor nestlings, were found to contain trypanosomatids in their intestine. Trypanosomes from the raptors and blood-sucking insects were isolated, and their 18S rRNA sequences were used for species identification and for the inference of intra- and interspecific relationships. Together with the trypanosome isolated from a black fly, the bird trypanosomes formed a well-supported Trypanosoma avium clade. The isolates derived from hippoboscid flies and mosquitoes are most likely also avian trypanosomes infecting birds other than the studied raptors. Analysis of the kinetoplast, that has features characteristic for the avian trypanosomes (minicircle size; dimensions of the kinetoplast disc), provided further evidence for the identification of vectors. It is suggested that all trypanosomes isolated from raptors included in this study belong to the T. avium complex and are transmitted by the ornithophilic simuliids such as Eusimulium securiforme.

  15. Heterogeneous infectiousness in guinea pigs experimentally infected with Trypanosoma cruzi.

    Science.gov (United States)

    Castillo-Neyra, Ricardo; Borrini Mayorí, Katty; Salazar Sánchez, Renzo; Ancca Suarez, Jenny; Xie, Sherrie; Náquira Velarde, Cesar; Levy, Michael Z

    2016-02-01

    Guinea pigs are important reservoirs of Trypanosoma cruzi, the causative parasite of Chagas disease, and in the Southern Cone of South America, transmission is mediated mainly by the vector Triatoma infestans. Interestingly, colonies of Triatoma infestans captured from guinea pig corrals sporadically have infection prevalence rates above 80%. Such high values are not consistent with the relatively short 7-8 week parasitemic period that has been reported for guinea pigs in the literature. We experimentally measured the infectious periods of a group of T. cruzi-infected guinea pigs by performing xenodiagnosis and direct microscopy each week for one year. Another group of infected guinea pigs received only direct microscopy to control for the effect that inoculation by triatomine saliva may have on parasitemia in the host. We observed infectious periods longer than those previously reported in a number of guinea pigs from both the xenodiagnosis and control groups. While some guinea pigs were infectious for a short time, other "super-shedders" were parasitemic up to 22 weeks after infection, and/or positive by xenodiagnosis for a year after infection. This heterogeneity in infectiousness has strong implications for T. cruzi transmission dynamics and control, as super-shedder guinea pigs may play a disproportionate role in pathogen spread. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  16. Crystal structure of arginine methyltransferase 6 from Trypanosoma brucei.

    Directory of Open Access Journals (Sweden)

    Chongyuan Wang

    Full Text Available Arginine methylation plays vital roles in the cellular functions of the protozoan Trypanosoma brucei. The T. brucei arginine methyltransferase 6 (TbPRMT6 is a type I arginine methyltransferase homologous to human PRMT6. In this study, we report the crystal structures of apo-TbPRMT6 and its complex with the reaction product S-adenosyl-homocysteine (SAH. The structure of apo-TbPRMT6 displays several features that are different from those of type I PRMTs that were structurally characterized previously, including four stretches of insertion, the absence of strand β15, and a distinct dimerization arm. The comparison of the apo-TbPRMT6 and SAH-TbPRMT6 structures revealed the fine rearrangements in the active site upon SAH binding. The isothermal titration calorimetry results demonstrated that SAH binding greatly increases the affinity of TbPRMT6 to a substrate peptide derived from bovine histone H4. The western blotting and mass spectrometry results revealed that TbPRMT6 methylates bovine histone H4 tail at arginine 3 but cannot methylate several T. brucei histone tails. In summary, our results highlight the structural differences between TbPRMT6 and other type I PRMTs and reveal that the active site rearrangement upon SAH binding is important for the substrate binding of TbPRMT6.

  17. Telomeric expression sites are highly conserved in Trypanosoma brucei.

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    Christiane Hertz-Fowler

    Full Text Available Subtelomeric regions are often under-represented in genome sequences of eukaryotes. One of the best known examples of the use of telomere proximity for adaptive purposes are the bloodstream expression sites (BESs of the African trypanosome Trypanosoma brucei. To enhance our understanding of BES structure and function in host adaptation and immune evasion, the BES repertoire from the Lister 427 strain of T. brucei were independently tagged and sequenced. BESs are polymorphic in size and structure but reveal a surprisingly conserved architecture in the context of extensive recombination. Very small BESs do exist and many functioning BESs do not contain the full complement of expression site associated genes (ESAGs. The consequences of duplicated or missing ESAGs, including ESAG9, a newly named ESAG12, and additional variant surface glycoprotein genes (VSGs were evaluated by functional assays after BESs were tagged with a drug-resistance gene. Phylogenetic analysis of constituent ESAG families suggests that BESs are sequence mosaics and that extensive recombination has shaped the evolution of the BES repertoire. This work opens important perspectives in understanding the molecular mechanisms of antigenic variation, a widely used strategy for immune evasion in pathogens, and telomere biology.

  18. Exosome secretion affects social motility in Trypanosoma brucei.

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    Dror Eliaz

    2017-03-01

    Full Text Available Extracellular vesicles (EV secreted by pathogens function in a variety of biological processes. Here, we demonstrate that in the protozoan parasite Trypanosoma brucei, exosome secretion is induced by stress that affects trans-splicing. Following perturbations in biogenesis of spliced leader RNA, which donates its spliced leader (SL exon to all mRNAs, or after heat-shock, the SL RNA is exported to the cytoplasm and forms distinct granules, which are then secreted by exosomes. The exosomes are formed in multivesicular bodies (MVB utilizing the endosomal sorting complexes required for transport (ESCRT, through a mechanism similar to microRNA secretion in mammalian cells. Silencing of the ESCRT factor, Vps36, compromised exosome secretion but not the secretion of vesicles derived from nanotubes. The exosomes enter recipient trypanosome cells. Time-lapse microscopy demonstrated that cells secreting exosomes or purified intact exosomes affect social motility (SoMo. This study demonstrates that exosomes are delivered to trypanosome cells and can change their migration. Exosomes are used to transmit stress signals for communication between parasites.

  19. Meiosis and haploid gametes in the pathogen Trypanosoma brucei.

    Science.gov (United States)

    Peacock, Lori; Bailey, Mick; Carrington, Mark; Gibson, Wendy

    2014-01-20

    In eukaryote pathogens, sex is an important driving force in spreading genes for drug resistance, pathogenicity, and virulence. For the parasitic trypanosomes that cause African sleeping sickness, mating occurs during transmission by the tsetse vector and involves meiosis, but haploid gametes have not yet been identified. Here, we show that meiosis is a normal part of development in the insect salivary glands for all subspecies of Trypanosoma brucei, including the human pathogens. By observing insect-derived trypanosomes during the window of peak expression of meiosis-specific genes, we identified promastigote-like (PL) cells that interacted with each other via their flagella and underwent fusion, as visualized by the mixing of cytoplasmic red and green fluorescent proteins. PL cells had a short, wide body, a very long anterior flagellum, and either one or two kinetoplasts, but only the anterior kinetoplast was associated with the flagellum. Measurement of nuclear DNA contents showed that PL cells were haploid relative to diploid metacyclics. Trypanosomes are among the earliest diverging eukaryotes, and our results support the hypothesis that meiosis and sexual reproduction are ubiquitous in eukaryotes and likely to have been early innovations. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  20. mRNA localization mechanisms in Trypanosoma cruzi.

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    Lysangela R Alves

    Full Text Available Asymmetric mRNA localization is a sophisticated tool for regulating and optimizing protein synthesis and maintaining cell polarity. Molecular mechanisms involved in the regulated localization of transcripts are widespread in higher eukaryotes and fungi, but not in protozoa. Trypanosomes are ancient eukaryotes that branched off early in eukaryote evolution. We hypothesized that these organisms would have basic mechanisms of mRNA localization. FISH assays with probes against transcripts coding for proteins with restricted distributions showed a discrete localization of the mRNAs in the cytoplasm. Moreover, cruzipain mRNA was found inside reservosomes suggesting new unexpected functions for this vacuolar organelle. Individual mRNAs were also mobilized to RNA granules in response to nutritional stress. The cytoplasmic distribution of these transcripts changed with cell differentiation, suggesting that localization mechanisms might be involved in the regulation of stage-specific protein expression. Transfection assays with reporter genes showed that, as in higher eukaryotes, 3'UTRs were responsible for guiding mRNAs to their final location. Our results strongly suggest that Trypanosoma cruzi have a core, basic mechanism of mRNA localization. This kind of controlled mRNA transport is ancient, dating back to early eukaryote evolution.

  1. Trypanosoma cruzi. Surface antigens of blood and culture forms

    International Nuclear Information System (INIS)

    Nogueira, N.; Chaplan, S.; Tydings, J.D.; Unkeless, J.; Cohn, Z.

    1981-01-01

    The surface polypeptides of both cultured and blood forms of Trypanosoma cruzi were iodinated by the glucose oxidase-lactoperoxidase technique. Blood-form trypomastigotes (BFT) isolated form infected mice displayed a major 90,000-Mr component. In contrast, both epimastigotes and trypomastigotes obtained form acellular cultures expressed a smaller 75,000-Mr peptide. Both major surface components were presumably glycoproteins in terms of their binding to concanavalin A-Sepharose 4B. Within a 3-h period, both blood and culture forms synthesized their respective surface glycoproteins (90,000 Mr and 75,000 Mr, respectively in vitro. [/sub 35/S]methionine-labeled surface peptides were immunoprecipitated with immune sera of both human and murine origin. A panel of sera form patients with chronic Chagas' disease and hyperimmunized mice recognized similar surface peptides. These immunogens were the same components as the major iodinated species. The major BFT surface peptide was readily removed by trypsin treatment of the parasites, although the procedure did not affect the 75,000-Mr peptide from the culture forms. Two-dimensional polyacrylamide gel electrophoresis revealed that the 90,000-Mr peptide found on BFT was an acidic protein of isoelectric point (pI) 5.0, whereas, the 75,000-Mr peptide form culture-form trypomastigotes has a pI of 7.2. The 90,000-Mr component is thought to be responsible for the anti-phagocytic properties of the BFT

  2. Early Trypanosoma cruzi Infection Reprograms Human Epithelial Cells

    Directory of Open Access Journals (Sweden)

    María Laura Chiribao

    2014-01-01

    Full Text Available Trypanosoma cruzi, the causative agent of Chagas disease, has the peculiarity, when compared with other intracellular parasites, that it is able to invade almost any type of cell. This property makes Chagas a complex parasitic disease in terms of prophylaxis and therapeutics. The identification of key host cellular factors that play a role in the T. cruzi invasion is important for the understanding of disease pathogenesis. In Chagas disease, most of the focus is on the response of macrophages and cardiomyocytes, since they are responsible for host defenses and cardiac lesions, respectively. In the present work, we studied the early response to infection of T. cruzi in human epithelial cells, which constitute the first barrier for establishment of infection. These studies identified up to 1700 significantly altered genes regulated by the immediate infection. The global analysis indicates that cells are literally reprogrammed by T. cruzi, which affects cellular stress responses (neutrophil chemotaxis, DNA damage response, a great number of transcription factors (including the majority of NFκB family members, and host metabolism (cholesterol, fatty acids, and phospholipids. These results raise the possibility that early host cell reprogramming is exploited by the parasite to establish the initial infection and posterior systemic dissemination.

  3. Cancer in the parasitic protozoans Trypanosoma brucei and Toxoplasma gondii.

    Science.gov (United States)

    Lun, Zhao-Rong; Lai, De-Hua; Wen, Yan-Zi; Zheng, Ling-Ling; Shen, Ji-Long; Yang, Ting-Bo; Zhou, Wen-Liang; Qu, Liang-Hu; Hide, Geoff; Ayala, Francisco J

    2015-07-21

    Cancer is a general name for more than 100 malignant diseases. It is postulated that all cancers start from a single abnormal cell that grows out of control. Untreated cancers can cause serious consequences and deaths. Great progress has been made in cancer research that has significantly improved our knowledge and understanding of the nature and mechanisms of the disease, but the origins of cancer are far from being well understood due to the limitations of suitable model systems and to the complexities of the disease. In view of the fact that cancers are found in various species of vertebrates and other metazoa, here, we suggest that cancer also occurs in parasitic protozoans such as Trypanosoma brucei, a blood parasite, and Toxoplasma gondii, an obligate intracellular pathogen. Without treatment, these protozoan cancers may cause severe disease and death in mammals, including humans. The simpler genomes of these single-cell organisms, in combination with their complex life cycles and fascinating life cycle differentiation processes, may help us to better understand the origins of cancers and, in particular, leukemias.

  4. Production of amastigotes from metacyclic trypomastigotes of Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Víctor T Contreras

    2002-12-01

    Full Text Available Attempts to recreate all the developmental stages of Trypanosoma cruzi in vitro have thus far been met with partial success. It is possible, for instance, to produce trypomastigotes in tissue culture and to obtain metacyclic trypomastigotes in axenic conditions. Even though T. cruzi amastigotes are known to differentiate from trypomastigotes and metacyclic trypomastigotes, it has only been possible to generate amastigotes in vitro from the tissue-culture-derived trypomastigotes. The factors and culture conditions required to trigger the transformation of metacyclic trypomastigotes into amastigotes are as yet undetermined. We show here that pre-incubation of metacyclic trypomastigotes in culture (MEMTAU medium at 37°C for 48 h is sufficient to commit the parasites to the transformation process. After 72 h of incubation in fresh MEMTAU medium, 90% of the metacyclic parasites differentiate into forms that are morphologically indistinguishable from normal amastigotes. SDS-PAGE, Western blot and PAABS analyses indicate that the transformation of axenic metacyclic trypomastigotes to amastigotes is associated with protein, glycoprotein and antigenic modifications. These data suggest that (a T. cruzi amastigotes can be obtained axenically in large amounts from metacyclic trypomastigotes, and (b the amastigotes thus obtained are morphological, biological and antigenically similar to intracellular amastigotes. Consequently, this experimental system may facilitate a direct, in vitro assessment of the mechanisms that enable T. cruzi metacyclic trypomastigotes to transform into amastigotes in the cells of mammalian hosts.

  5. Geographical Distribution of Trypanosoma cruzi Genotypes in Venezuela

    Science.gov (United States)

    Carrasco, Hernán J.; Segovia, Maikell; Llewellyn, Martin S.; Morocoima, Antonio; Urdaneta-Morales, Servio; Martínez, Cinda; Martínez, Clara E.; Garcia, Carlos; Rodríguez, Marlenes; Espinosa, Raul; de Noya, Belkisyolé A.; Díaz-Bello, Zoraida; Herrera, Leidi; Fitzpatrick, Sinead; Yeo, Matthew; Miles, Michael A.; Feliciangeli, M. Dora

    2012-01-01

    Chagas disease is an endemic zoonosis native to the Americas and is caused by the kinetoplastid protozoan parasite Trypanosoma cruzi. The parasite is also highly genetically diverse, with six discrete typing units (DTUs) reported TcI – TcVI. These DTUs broadly correlate with several epidemiogical, ecological and pathological features of Chagas disease. In this manuscript we report the most comprehensive evaluation to date of the genetic diversity of T. cruzi in Venezuela. The dataset includes 778 samples collected and genotyped over the last twelve years from multiple hosts and vectors, including nine wild and domestic mammalian host species, and seven species of triatomine bug, as well as from human sources. Most isolates (732) can be assigned to the TcI clade (94.1%); 24 to the TcIV group (3.1%) and 22 to TcIII (2.8%). Importantly, among the 95 isolates genotyped from human disease cases, 79% belonged to TcI - a DTU common in the Americas, however, 21% belonged to TcIV- a little known genotype previously thought to be rare in humans. Furthermore, were able to assign multiple oral Chagas diseases cases to TcI in the area around the capital, Caracas. We discuss our findings in the context of T. cruzi DTU distributions elsewhere in the Americas, and evaluate the impact they have on the future of Chagas disease control in Venezuela. PMID:22745843

  6. The Oral Antimalarial Drug Tafenoquine Shows Activity against Trypanosoma brucei.

    Science.gov (United States)

    Carvalho, Luis; Martínez-García, Marta; Pérez-Victoria, Ignacio; Manzano, José Ignacio; Yardley, Vanessa; Gamarro, Francisco; Pérez-Victoria, José M

    2015-10-01

    The protozoan parasite Trypanosoma brucei causes human African trypanosomiasis, or sleeping sickness, a neglected tropical disease that requires new, safer, and more effective treatments. Repurposing oral drugs could reduce both the time and cost involved in sleeping sickness drug discovery. Tafenoquine (TFQ) is an oral antimalarial drug belonging to the 8-aminoquinoline family which is currently in clinical phase III. We show here that TFQ efficiently kills different T. brucei spp. in the submicromolar concentration range. Our results suggest that TFQ accumulates into acidic compartments and induces a necrotic process involving cell membrane disintegration and loss of cytoplasmic content, leading to parasite death. Cell lysis is preceded by a wide and multitarget drug action, affecting the lysosome, mitochondria, and acidocalcisomes and inducing a depolarization of the mitochondrial membrane potential, elevation of intracellular Ca(2+), and production of reactive oxygen species. This is the first report of an 8-aminoquinoline demonstrating significant in vitro activity against T. brucei. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  7. Effects of azadirachtin on Rhodnius prolixus: immunity and trypanosoma interaction

    Directory of Open Access Journals (Sweden)

    Patricia de Azambuja

    1992-01-01

    Full Text Available The effects of azadirachtin, a tetranortriterpenoid from the neem tree Aradirachta indica J. on both immunity and Trypanosoma cruzi interaction within Rhodniusprolixus and other triatomines, were presented Given through a blood meal, azadirachtin affected the immune reactivity as shown by a significant reduction in numbers of hemocytes and consequently nodule formation follwing challenge with Enterobacter cloacae ß12, reduction in ability to produce antibacterial activities in the hemolymph when injected with bacteria, and decreased ability to destroy the infection caused by inoculation of E. cloacae cells. A single dose of azadirachtin was able to block the development of T. cruzi in R. prolixus if given through the meal at different intervals, together with, before or after parasite infection. Similary, these results were observed with different triatomine species and different strains of T. cruzi. Azadirachtin induced a permanent resistance of the vector against reinfection with T. cruzi. The significance of these data is discussed in relation to the general mode of azadirachtin action in insects.

  8. In vitro effects of citral on Trypanosoma cruzi metacyclogenesis

    Directory of Open Access Journals (Sweden)

    Josiane Cardoso

    2010-12-01

    Full Text Available Citral, the main constituent of lemongrass (Cymbopogon citratus essential oil, was added to Trypanosoma cruzi cultures grown in TAU3AAG medium to observe the effect on the epimastigote-to-trypomastigote differentiation process (metacyclogenesis. Our results showed that citral (20 μg/mL did not affect epimastigote viability or inhibit the differentiation process. Concentrations higher than 60 μg/mL, however, led to 100% cell death (both epimastigote and trypomastigote forms. Although epimastigotes incubated with 30 μg/mL citral were viable and able to adhere to the substrate, we observed around 50% inhibition in metacyclogenesis, with a calculated concentration that inhibited metacyclogenesis by 50% after 24 h (IC50/24 h of about 31 μg/mL. Treatment with 30 μg/mL citral did not hinder epimastigote multiplication because epimastigote growth resumed when treated cells were transferred to a drug-free liver infusion tryptose culture medium. Metacyclogenesis was almost totally abolished at 40 μg/mL after 24 h of incubation. Furthermore, the metacyclic trypomastigotes obtained in vitro were similarly susceptible to citral, with an IC50/24 h, concentration that killed 50% of the cells after 24 h, of about 24.5 μg/mL. Therefore, citral appears to be a good candidate as an inhibitory drug for further studies analyzing the T. cruzi metacyclogenesis process.

  9. Lysophosphatidylcholine: A Novel Modulator of Trypanosoma cruzi Transmission

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    Mário A. C. Silva-Neto

    2012-01-01

    Full Text Available Lysophosphatidylcholine is a bioactive lipid that regulates a large number of cellular processes and is especially present during the deposition and infiltration of inflammatory cells and deposition of atheromatous plaque. Such molecule is also present in saliva and feces of the hematophagous organism Rhodnius prolixus, a triatominae bug vector of Chagas disease. We have recently demonstrated that LPC is a modulator of Trypanosoma cruzi transmission. It acts as a powerful chemoattractant for inflammatory cells at the site of the insect bite, which will provide a concentrated population of cells available for parasite infection. Also, LPC increases macrophage intracellular calcium concentrations that ultimately enhance parasite invasion. Finally, LPC inhibits NO production by macrophages stimulated by live T. cruzi, and thus interferes with the immune system of the vertebrate host. In the present paper, we discuss the main signaling mechanisms that are likely used by such molecule and their eventual use as targets to block parasite transmission and the pathogenesis of Chagas disease.

  10. Seropositivity for Trypanosoma cruzi in domestic dogs from Sonora, Mexico.

    Science.gov (United States)

    Arce-Fonseca, Minerva; Carrillo-Sánchez, Silvia C; Molina-Barrios, Ramón M; Martínez-Cruz, Mariana; Cedillo-Cobián, Jesús R; Henao-Díaz, Yuly A; Rodríguez-Morales, Olivia

    2017-09-05

    Chagas disease is an important health problem in Latin America due to its incapacitating effects and associated mortality. Studies on seropositivity for Trypanosoma cruzi in Mexican dogs have demonstrated a direct correlation between seropositivity in humans and dogs, which can act as sentinels for the disease in this region. The objective of this study was to determine the seropositivity for T.cruzi infection in dogs from Sonora, a northern borderstate of Mexico. Responsible pet owners were selected at random from an urban area of Empalme municipality, Sonora, Mexico, and from there, 180 dog samples were collected. Anti-T. cruzi antibodies were determined using the enzyme-linked immunosorbent assay (ELISA) method. Reactive ELISA sera were processed by indirect immunofluorescence to confirm the presence of anti-T. cruzi antibodies. For the statistical analysis, chi-square tests were conducted. Dogs' sera showed a seropositivity rate of 4.44%. The rate of seropositivity was not associated with the dogs' age, sex, or socioeconomics pertaining to the geographical area. One sample (1/180, 0.55%) showed the acute state of the disease. The study found a presence of anti-T. cruzi antibodies in dogs in this area, which suggests vector transmission. There is a need for active surveillance programs throughout the state of Sonora and vector control strategies should also be implemented in endemic regions.

  11. Diterpenoids from Azorella compacta (Umbelliferae active on Trypanosoma cruzi

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    Araya Jorge E

    2003-01-01

    Full Text Available The anti-Trypanosoma cruzi activity of natural products isolated from Azorella compacta was evaluated, with particular emphasis on their effect against intracellular amastigotes. Five diterpenoids from A. compacta derived from mulinane and azorellane were isolated and identified. Only two products, named azorellanol (Y-2 and mulin-11,3-dien-20-oic acid (Y-5, showed trypanocidal activity against all stages of T. cruzi including intracellular amastigotes. At 10 µM, these compounds displayed a strong lytic activity. It ranged from 88.4 ± 0.6 to 99.0 ± 1 % for all strains and stages evaluate, with an IC50 /18 h values of 20-84 µM and 41-87 µM, respectively. The development of intracellular amastigotes was also inhibited by nearly 60% at 25 µM. The trypanocidal molecules Y-2 and Y-5 did show different degrees of cytotoxicity depending on the cell line tested, with an IC50 /24 h ranging from 33.2 to 161.2 µM. We evaluated the effect of diterpenoids against intracellular T. cruzi forms by immunofluorescent identification of a specific membrane molecular marker (Ssp-4 antigen of the T. cruzi amastigote forms. The accuracy and reproducibility of the measurements were found to be outstanding when examined by confocal microscopy.

  12. The activity of aminoglycoside antibiotics against Trypanosoma brucei.

    Science.gov (United States)

    Maina, N W; Kinyanjui, B; Onyango, J D; Auma, J E; Croj, S

    1998-01-01

    The trypanocidal activity of four aminoglycosides was determined against Trypanosoma brucei in vitro. The drug activity in descending order, was as follows; paromomycin kanamycin>gentamycin > neomycin. Paromomycin bad the highest activity and the concentration that inhibited 50% of trypanosome growth (IC50) was 11.4microM. The effect of paromomycin on the causative agents of the East African form of sleeping sickness - T.b. rhodesiense KETRI 265, 2285, 2545, 2562 and EATRO 110,112, 1152 was subsequently assessed. Variations sensitivities between the trypanosome populations were observed and IC50 values ranging from 13.01 to 43.06 microM recorded. However, when paromomycin was administered intraperitoneally (i.p) at 500 mg/kg, it was not effective in curing mice infected with T. b. rhodesienseKETRI 2545 the most drug-sensitive isolate in vitro. Lack of in vivo activity may be because the trypanosome is an extracellular parasite. The pharmacokinetics of paromomycin in the mouse model need to be determined.

  13. The morphology of ovine Trypanosoma melophagium (zoomastigophorea: kinetoplastida).

    Science.gov (United States)

    Büscher, G; Friedhoff, K T

    1984-02-01

    Morphologic and biometric data on bloodstream stages of Trypanosoma melophagium are presented. An increasing parasitemia with 111 trypomastigote stages of T. melophagium were found in Giemsa-stained thin blood smears taken from a splenectomized, cortisone-treated sheep recently infested with Melophagus ovinus infected with T. melophagium . The arithmetic mean and standard deviation in micron of the distances between posterior end and kinetoplast were 14.7 and 2.9, from the kinetoplastic to the center of the nucleus 5.1 and 1.1, and from there to the anterior end 19.5 and 1.9. The free flagellum measured 6.0 microns +/- 1.6 microns. The median and the range of the central 70% of values (median +/- 35%) of the nuclear index were 1.1 and 0.9-1.2 and of the kinetoplastic index 3.8 and 3.3-4.9. The same data in microns for the maximal width were 3.1 and 2.1-4.6, and for the width at the level of the nucleus 2.9 and 2.2-4.6. The larger and smaller diameters of the nucleus measured 2.6 (2.2-3.7) micron and 1.7 (1.3-1.7) micron, respectively. The corresponding kinetoplast diameters were 1.1 (0.9-1.3) microns and 0.9 (0.6-0.9) micron, respectively.

  14. Predominance of Trypanosoma rangeli infection in children from a Chagas disease endemic area in the west-shore of the Panama canal

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    Azael Saldaña

    2005-11-01

    Full Text Available A total of 206 serum samples from children (3-14 years old living in the Amador County (La Chorrera District, Province of Panama were screened by indirect immunofluorescence antibody test (IFAT for the presence of antibodies against Trypanosoma cruzi. Positive sera were confirmed by recombinant enzyme-linked immunosorbent assay (ELISA and Western blot analysis. The presence of blood trypanosomes was investigated by hemoculture and subsequently identify by a duplex polymerase chain reaction (PCR followed by dot blot hybridization. The results indicated a prevalence of 9.7% for trypanosome infections, a seroprevalence of 2.9% against T. cruzi and a predominance of T. rangeli infection (6.8%. The immunological and clinical implications of these findings are discussed.

  15. Rapid Parallel Screening for Strain Optimization

    Science.gov (United States)

    2013-08-16

    fermentation yields of industrially relevant biological compounds. Screening of the desired chemicals was completed previously. Microbes that can...reporter, and, 2) a yeast TAR cloning shuttle vector for transferring catabolic clusters to E. coli. 15. SUBJECT TERMS NA 16. SECURITY CLASSIFICATION OF... fermentation yields of industrially relevant biological compounds. Screening of the desired chemicals was completed previously. Microbes that can utilize

  16. Trypanosoma cruzi: vertebrate and invertebrate cycles in the same mammal host, the opossum Didelphis marsupialis

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    Maria P. Deane

    1984-12-01

    Full Text Available Epimastigotes multiplying extracellularly and metacyclic trypomastigotes, stages that correspond to the cycle of Trypanosoma cruzi in the intestinal lumen of its insect vector, were consistently found in the lumen of the anal glands of opossums Didelphis marsupialis inoculated subcutaneously with infective feces of triatomid bugs.No gambá (Didelphis marsupialis foi observado um ciclo extracelular do Trypanosoma cruzi: o parasita crescia abundantemente no material de secreção acumulado no lumen das glandulas anais de animais criados em cativeiro e infectados por via subcutanea com fezes de triatomineos.

  17. Toxicology screen

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003578.htm Toxicology screen To use the sharing features on this page, please enable JavaScript. A toxicology screen refers to various tests that determine the ...

  18. Yield stress fluids slowly yield to analysis

    NARCIS (Netherlands)

    Bonn, D.; Denn, M.M.

    2009-01-01

    We are surrounded in everyday life by yield stress fluids: materials that behave as solids under small stresses but flow like liquids beyond a critical stress. For example, paint must flow under the brush, but remain fixed in a vertical film despite the force of gravity. Food products (such as

  19. Screening for skin cancer.

    Science.gov (United States)

    Helfand, M; Mahon, S M; Eden, K B; Frame, P S; Orleans, C T

    2001-04-01

    Malignant melanoma is often lethal, and its incidence in the United States has increased rapidly over the past 2 decades. Nonmelanoma skin cancer is seldom lethal, but, if advanced, can cause severe disfigurement and morbidity. Early detection and treatment of melanoma might reduce mortality, while early detection and treatment of nonmelanoma skin cancer might prevent major disfigurement and to a lesser extent prevent mortality. Current recommendations from professional societies regarding screening for skin cancer vary. To examine published data on the effectiveness of routine screening for skin cancer by a primary care provider, as part of an assessment for the U.S. Preventive Services Task Force. We searched the MEDLINE database for papers published between 1994 and June 1999, using search terms for screening, physical examination, morbidity, and skin neoplasms. For information on accuracy of screening tests, we used the search terms sensitivity and specificity. We identified the most important studies from before 1994 from the Guide to Clinical Preventive Services, second edition, and from high-quality reviews. We used reference lists and expert recommendations to locate additional articles. Two reviewers independently reviewed a subset of 500 abstracts. Once consistency was established, the remainder were reviewed by one reviewer. We included studies if they contained data on yield of screening, screening tests, risk factors, risk assessment, effectiveness of early detection, or cost effectiveness. We abstracted the following descriptive information from full-text published studies of screening and recorded it in an electronic database: type of screening study, study design, setting, population, patient recruitment, screening test description, examiner, advertising targeted at high-risk groups or not targeted, reported risk factors of participants, and procedure for referrals. We also abstracted the yield of screening data including probabilities and numbers

  20. Serosurvey for Leishmania spp., Toxoplasma gondii, Trypanosoma cruzi and Neospora caninum in neighborhood dogs in Curitiba-Paraná, Brazil

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    Caroline Constantino

    Full Text Available Abstract Neighborhood dogs may act as reservoirs for several zoonotic protozoan infections, particularly in urban areas, thus constituting a potential public health threat. Accordingly, the aim of the present study was to evaluate the exposure of neighborhood dogs to four protozoan pathogens in public areas with high levels of human movement in Curitiba, southern Brazil. Blood samples from 26 neighborhood dogs were screened by means of the indirect immunofluorescent antibody test (IFAT for Leishmania spp., Toxoplasma gondii, Trypanosoma cruzi and Neospora caninum, and a questionnaire was answered by the respective keeper. A total of 8/26 dogs (30.7% seroreactive to T. gondii, 3/26 (11.5% to N. caninum and 2/26 (7.7% to both were identified. All the samples were seronegative for T. cruzi and Leishmania spp. Pathogen seroreactivity was not associated with the daily human movements or other epidemiological variables investigated (p > 0.05. In conclusion, the low seroprevalence for T. gondii and N. caninum indicated low environmental and food risk for animal infection and the seronegativity for Leishmania spp. and T. cruzi may reflect the absence of these pathogens in urban areas of Curitiba. Moreover, neighborhood dogs may be used as environmental sentinels for the presence of protozoan pathogens and their vectors.

  1. Bond yield curve construction

    Directory of Open Access Journals (Sweden)

    Kožul Nataša

    2014-01-01

    Full Text Available In the broadest sense, yield curve indicates the market's view of the evolution of interest rates over time. However, given that cost of borrowing it closely linked to creditworthiness (ability to repay, different yield curves will apply to different currencies, market sectors, or even individual issuers. As government borrowing is indicative of interest rate levels available to other market players in a particular country, and considering that bond issuance still remains the dominant form of sovereign debt, this paper describes yield curve construction using bonds. The relationship between zero-coupon yield, par yield and yield to maturity is given and their usage in determining curve discount factors is described. Their usage in deriving forward rates and pricing related derivative instruments is also discussed.

  2. Measurements of fission yields

    International Nuclear Information System (INIS)

    Denschlag, H.O.

    2000-01-01

    After some historical introductory remarks on the discovery of nuclear fission and early fission yield determinations, the present status of knowledge on fission yields is briefly reviewed. Practical and fundamental reasons motivating the pursuit of fission yield measurements in the coming century are pointed out. Recent results and novel techniques are described that promise to provide new interesting insights into the fission process during the next century. (author)

  3. Active transcription and ultrastructural changes during Trypanosoma cruzi metacyclogenesis

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    Ludmila R.P. Ferreira

    2008-03-01

    Full Text Available The differentiation of proliferating epimastigote forms of Trypanosoma cruzi , the protozoan parasite that causes Chagas’ disease, into the infective and non-proliferating metacyclic forms can be reproduced in the laboratory by incubating the cells in a chemically-defined medium that mimics the urine of the insect vector. Epimastigotes have a spherical nucleus, a flagellum protruding from the middle of the protozoan cell, and a disk-shaped kinetoplast - an organelle that corresponds to the mitochondrial DNA. Metacyclic trypomastigotes have an elongated shape with the flagellum protruding from the posterior portion of the cell and associated with a spherical kinetoplast. Here we describe the morphological events of this transformation and characterize a novel intermediate stage by three-dimensional reconstruction of electron microscope serial sections. This new intermediate stage is characterized by a kinetoplast compressing an already elongated nucleus, indicating that metacyclogenesis involves active movements of the flagellar structure relative to the cell body. As transcription occurs more intensely in proliferating epimastigotes than in metacyclics, we also examined the presence of RNA polymerase II and measured transcriptional activity during the differentiation process. Both the presence of the enzyme and transcriptional activity remain unchanged during all steps of metacyclogenesis. RNA polymerase II levels and transcriptional activity only decrease after metacyclics are formed. We suggest that transcription is required during the epimastigote-to-metacyclic trypomastigote differentiation process, until the kinetoplast and flagellum reach the posterior position of the parasites in the infective form.A diferenciação de formas epimastigotas (proliferativas do Trypanosoma cruzi, parasita protozoário causador da doença de Chagas, em formas metacíclicas tripomastigotas (infectivas e não proliferativas, pode ser reproduzida em laborat

  4. Molecular basis of mammalian cell invasion by Trypanosoma cruzi

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    Nobuko Yoshida

    2006-03-01

    Full Text Available Establishment of infection by Trypanosoma cruzi, the agent of Chagas' disease, depends on a series of events involving interactions of diverse parasite molecules with host components. Here we focus on the mechanisms of target cell invasion by metacyclic trypomastigotes (MT and mammalian tissue culture trypomastigotes (TCT. During MT or TCT internalization, signal transduction pathways are activated both in the parasite and the target cell, leading to Ca2+ mobilization. For cell adhesion, MT engage surface glycoproteins, such as gp82 and gp35/50, which are Ca2+ signal-inducing molecules. In T. cruzi isolates that enter host cells in gp82-mediated manner, parasite protein tyrosine kinase as well as phospholipase C are activated, and Ca2+ is released from I P3-sensitive stores, whereas in T. cruzi isolates that attach to target cells mainly through gp35/50, the signaling pathway involving adenylate cyclase appears to be stimulated, with Ca2+ release from acidocalciosomes. In addition, T. cruzi isolate-dependent inhibitory signals, mediated by MT-specific gp90, may be triggered both in the host cell and the parasite. The repertoire of TCT molecules implicated in cell invasion includes surface glycoproteins of gp85 family, with members containing binding sites for laminin and cytokeratin 18, enzymes such as cruzipain, trans-sialidase, and an oligopeptidase B that generates a Ca2+-agonist from a precursor molecule.O estabelecimento da infecção por Trypanosoma cruzi, o agente da doença de Chagas, depende de uma série de eventos envolvendo interações de diversas moléculas do parasita com componentes do hospedeiro. Focalizamos aqui os mecanismos de invasão celular por tripomastigotas metacíclicos (TM e por tripomastigotas de cultura de tecido (TCT. Durante a internalização de TM ou TCT, vias de transdução de sinal são ativadas tanto no parasita como na célula alvo, acarretando a mobilização de Ca2+. Para adesão, TM utiliza as glicoprote

  5. Eco-epidemiological aspects of Trypanosoma cruzi, Trypanosoma rangeli and their vector (Rhodnius pallescens in Panama Generalidades do Trypanosoma cruzi, do Trypanosoma rangeli e do seu vetor (Rhodnius pallescens no Panamá

    Directory of Open Access Journals (Sweden)

    Ana Maria de Vasquez

    2004-08-01

    Full Text Available The eco-epidemiology of T. cruzi infection was investigated in the Eastern border of the Panama Canal in Central Panama. Between 1999 and 2000, 1110 triatomines were collected: 1050 triatomines (94.6% from palm trees, 27 (2.4% from periurban habitats and 33 (3.0% inside houses. All specimens were identified as R. pallescens. There was no evidence of vector domiciliation. Salivary glands from 380 R. pallescens revealed a trypanosome natural infection rate of 7.6%, while rectal ampoule content from 373 triatomines was 45%. Isoenzyme profiles on isolated trypanosomes demonstrated that 85.4% (n = 88 were T. cruzi and 14.6% (n = 15 were T. rangeli. Blood meal analysis from 829 R. pallescens demonstrated a zoophilic vector behavior, with opossums as the preferential blood source. Seroprevalence in human samples from both study sites was less than 2%. Our results demonstrate that T. cruzi survives in the area in balanced association with R. pallescens, and with several different species of mammals in their natural niches. However, the area is an imminent risk of infection for its population, consequently it is important to implement a community educational program regarding disease knowledge and control measures.A epidemiologia da infecção do T. cruzi foi investigada na margem oriental do canal do Panamá, na região central da Republica do Panamá. A informação obtida durante o estudo avaliou fatores de risco da doença de Chagas nesta área. Entre 1999 e 2000, 1110 triatomíneos foram coletados: 1050 triatomíneos (94,6% em palmeiras, 27 (2,4% em habitats periurbanos e 33 (3,0% no interior de casas. Todos os espécimens foram identificados como R. pallescens. Não havia nenhuma evidência de domiciliação do vetor. O exame de glândulas salivares de 380 R. pallescens revelaram taxa de infecção natural por Trypanosoma de 7,6%, mas o conteúdo da ampola rectal de 373 triatomíneos mostrou 45% de positividade. Os perfis de isoenzimas em

  6. Fission product yields

    International Nuclear Information System (INIS)

    Valenta, V.; Hep, J.

    1978-01-01

    Data are summed up necessary for determining the yields of individual fission products from different fissionable nuclides. Fractional independent yields, cumulative and isobaric yields are presented here for the thermal fission of 235 U, 239 Pu, 241 Pu and for fast fission (approximately 1 MeV) of 235 U, 238 U, 239 Pu, 241 Pu; these values are included into the 5th version of the YIELDS library, supplementing the BIBFP library. A comparison is made of experimental data and possible improvements of calculational methods are suggested. (author)

  7. The regulation of autophagy differentially affects Trypanosoma cruzi metacyclogenesis.

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    María Cristina Vanrell

    2017-11-01

    Full Text Available Autophagy is a cellular process required for the removal of aged organelles and cytosolic components through lysosomal degradation. All types of eukaryotic cells from yeasts to mammalian cells have the machinery to activate autophagy as a result of many physiological and pathological situations. The most frequent stimulus of autophagy is starvation and the result, in this case, is the fast generation of utilizable food (e.g. amino acids and basic nutrients to maintain the vital biological processes. In some organisms, starvation also triggers other associated processes such as differentiation. The protozoan parasite Trypanosoma cruzi undergoes a series of differentiation processes throughout its complex life cycle. Although not all autophagic genes have been identified in the T. cruzi genome, previous works have demonstrated the presence of essential autophagic-related proteins. Under starvation conditions, TcAtg8, which is the parasite homolog of Atg8/LC3 in other organisms, is located in autophagosome-like vesicles. In this work, we have characterized the autophagic pathway during T. cruzi differentiation from the epimastigote to metacyclic trypomastigote form, a process called metacyclogenesis. We demonstrated that autophagy is stimulated during metacyclogenesis and that the induction of autophagy promotes this process. Moreover, with exception of bafilomycin, other classical autophagy modulators have similar effects on T. cruzi autophagy. We also showed that spermidine and related polyamines can positively regulate parasite autophagy and differentiation. We concluded that both polyamine metabolism and autophagy are key processes during T. cruzi metacyclogenesis that could be exploited as drug targets to avoid the parasite cycle progression.

  8. Protein 3-nitrotyrosine formation during Trypanosoma cruzi infection in mice

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    M. Naviliat

    2005-12-01

    Full Text Available Nitric oxide (·NO is a diffusible messenger implicated in Trypanosoma cruzi resistance. Excess production of ·NO and oxidants leads to the generation of nitrogen dioxide (·NO2, a strong nitrating agent. Tyrosine nitration is a post-translational modification resulting from the addition of a nitro (-NO2 group to the ortho-position of tyrosine residues. Detection of protein 3-nitrotyrosine is regarded as a marker of nitro-oxidative stress and is observed in inflammatory processes. The formation and role of nitrating species in the control and myocardiopathy of T. cruzi infection remain to be studied. We investigated the levels of ·NO and protein 3-nitrotyrosine in the plasma of C3H and BALB/c mice and pharmacologically modulated their production during the acute phase of T. cruzi infection. We also looked for protein 3-nitrotyrosine in the hearts of infected animals. Our results demonstrated that C3H animals produced higher amounts of ·NO than BALB/c mice, but their generation of peroxynitrite was not proportionally enhanced and they had higher parasitemias. While N G-nitro-arginine methyl ester treatment abolished ·NO production and drastically augmented the parasitism, mercaptoethylguanidine and guanido-ethyl disulfide, at doses that moderately reduced the ·NO and 3-nitrotyrosine levels, paradoxically diminished the parasitemia in both strains. Nitrated proteins were also demonstrated in myocardial cells of infected mice. These data suggest that the control of T. cruzi infection depends not only on the capacity to produce ·NO, but also on its metabolic fate, including the generation of nitrating species that may constitute an important element in parasite resistance and collateral myocardial damage.

  9. Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology.

    Directory of Open Access Journals (Sweden)

    Andrés B Lantos

    2016-04-01

    Full Text Available Trypanosoma cruzi, the flagellate protozoan agent of Chagas disease or American trypanosomiasis, is unable to synthesize sialic acids de novo. Mucins and trans-sialidase (TS are substrate and enzyme, respectively, of the glycobiological system that scavenges sialic acid from the host in a crucial interplay for T. cruzi life cycle. The acquisition of the sialyl residue allows the parasite to avoid lysis by serum factors and to interact with the host cell. A major drawback to studying the sialylation kinetics and turnover of the trypomastigote glycoconjugates is the difficulty to identify and follow the recently acquired sialyl residues. To tackle this issue, we followed an unnatural sugar approach as bioorthogonal chemical reporters, where the use of azidosialyl residues allowed identifying the acquired sugar. Advanced microscopy techniques, together with biochemical methods, were used to study the trypomastigote membrane from its glycobiological perspective. Main sialyl acceptors were identified as mucins by biochemical procedures and protein markers. Together with determining their shedding and turnover rates, we also report that several membrane proteins, including TS and its substrates, both glycosylphosphatidylinositol-anchored proteins, are separately distributed on parasite surface and contained in different and highly stable membrane microdomains. Notably, labeling for α(1,3Galactosyl residues only partially colocalize with sialylated mucins, indicating that two species of glycosylated mucins do exist, which are segregated at the parasite surface. Moreover, sialylated mucins were included in lipid-raft-domains, whereas TS molecules are not. The location of the surface-anchored TS resulted too far off as to be capable to sialylate mucins, a role played by the shed TS instead. Phosphatidylinositol-phospholipase-C activity is actually not present in trypomastigotes. Therefore, shedding of TS occurs via microvesicles instead of as a fully

  10. Prevalence of Trypanosoma vivax in cattle in central Sudan

    International Nuclear Information System (INIS)

    Fadl, M.; Babiker, H.I.; Bakheit, M.A.; A Rahman, A.H.

    2000-01-01

    The study was conducted to validate an antibody-detection ELISA test (Ab-ELISA) using pre-coated ELISA plates with crude antigen preparation of Trypanosoma vivax and to study the prevalence of T. vivax infection in central Sudan. A total of 704 blood samples were collected from cattle in central Sudan, a known endemic area of T. vivax infection. Additionally, 74 blood samples were collected from northern Sudan (Atbra town), an area presumed to be T. vivax-free. Sera were collected during the period September 1998 to May 1999 during three different seasons (summer, autumn and winter). Under the existing laboratory conditions, the test showed a clear distinction between different controls, i.e. strong positive control (C++), weak positive control (C+), negative control (C-) and the conjugate control (Cc). A percent positivity of 25% was taken as a cut-off value to determine the positivity or negativity of the test. The acceptable optical density range of strong positive control (C++) was 0.65-1.22. Lower and upper percent positivity limits for different controls were also determined. The study showed that T. vivax is endemic in central Sudan with 1.4% prevalence based on parasitological examination and 29.26% on Ab-ELISA. The infection rate was significantly higher during the autumn and winter than in summer. Young cattle showed significantly lower infection rates than adults as indicated by both the parasitological and the Ab-ELISA test. In relation to husbandry practice, migratory cattle showed significantly higher rates of prevalence than resident cattle. There was no significant difference in average packed red cell volume (PCV) values between ELISA positive and ELISA negative animals. Calves of less than one year of age showed significantly lower PCV values when belonging to migratory herds than to resident herds. (author)

  11. Functional characterization of 8-oxoguanine DNA glycosylase of Trypanosoma cruzi.

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    Carolina Furtado

    Full Text Available The oxidative lesion 8-oxoguanine (8-oxoG is removed during base excision repair by the 8-oxoguanine DNA glycosylase 1 (Ogg1. This lesion can erroneously pair with adenine, and the excision of this damaged base by Ogg1 enables the insertion of a guanine and prevents DNA mutation. In this report, we identified and characterized Ogg1 from the protozoan parasite Trypanosoma cruzi (TcOgg1, the causative agent of Chagas disease. Like most living organisms, T. cruzi is susceptible to oxidative stress, hence DNA repair is essential for its survival and improvement of infection. We verified that the TcOGG1 gene encodes an 8-oxoG DNA glycosylase by complementing an Ogg1-defective Saccharomyces cerevisiae strain. Heterologous expression of TcOGG1 reestablished the mutation frequency of the yeast mutant ogg1(-/- (CD138 to wild type levels. We also demonstrate that the overexpression of TcOGG1 increases T. cruzi sensitivity to hydrogen peroxide (H(2O(2. Analysis of DNA lesions using quantitative PCR suggests that the increased susceptibility to H(2O(2 of TcOGG1-overexpressor could be a consequence of uncoupled BER in abasic sites and/or strand breaks generated after TcOgg1 removes 8-oxoG, which are not rapidly repaired by the subsequent BER enzymes. This hypothesis is supported by the observation that TcOGG1-overexpressors have reduced levels of 8-oxoG both in the nucleus and in the parasite mitochondrion. The localization of TcOgg1 was examined in parasite transfected with a TcOgg1-GFP fusion, which confirmed that this enzyme is in both organelles. Taken together, our data indicate that T. cruzi has a functional Ogg1 ortholog that participates in nuclear and mitochondrial BER.

  12. Cynaropicrin targets the trypanothione redox system in Trypanosoma brucei.

    Science.gov (United States)

    Zimmermann, Stefanie; Oufir, Mouhssin; Leroux, Alejandro; Krauth-Siegel, R Luise; Becker, Katja; Kaiser, Marcel; Brun, Reto; Hamburger, Matthias; Adams, Michael

    2013-11-15

    In mice cynaropicrin (CYN) potently inhibits the proliferation of Trypanosoma brucei-the causative agent of Human African Trypanosomiasis-by a so far unknown mechanism. We hypothesized that CYNs α,β-unsaturated methylene moieties act as Michael acceptors for glutathione (GSH) and trypanothione (T(SH)2), the main low molecular mass thiols essential for unique redox metabolism of these parasites. The analysis of this putative mechanism and the effects of CYN on enzymes of the T(SH)2 redox metabolism including trypanothione reductase, trypanothione synthetase, glutathione-S-transferase, and ornithine decarboxylase are shown. A two step extraction protocol with subsequent UPLC-MS/MS analysis was established to quantify intra-cellular CYN, T(SH)2, GSH, as well as GS-CYN and T(S-CYN)2 adducts in intact T. b. rhodesiense cells. Within minutes of exposure to CYN, the cellular GSH and T(SH)2 pools were entirely depleted, and the parasites entered an apoptotic stage and died. CYN also showed inhibition of the ornithine decarboxylase similar to the positive control eflornithine. Significant interactions with the other enzymes involved in the T(SH)2 redox metabolism were not observed. Alongside many other biological activities sesquiterpene lactones including CYN have shown antitrypanosomal effects, which have been postulated to be linked to formation of Michael adducts with cellular nucleophiles. Here the interaction of CYN with biological thiols in a cellular system in general, and with trypanosomal T(SH)2 redox metabolism in particular, thus offering a molecular explanation for the antitrypanosomal activity is demonstrated. At the same time, the study provides a novel extraction and analysis protocol for components of the trypanosomal thiol metabolism. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Rab23 is a flagellar protein in Trypanosoma brucei

    Directory of Open Access Journals (Sweden)

    Field Mark C

    2011-06-01

    Full Text Available Abstract Background Rab small GTPases are important mediators of membrane transport, and orthologues frequently retain similar locations and functions, even between highly divergent taxa. In metazoan organisms Rab23 is an important negative regulator of Sonic hedgehog signaling and is crucial for correct development and differentiation of cellular lineages by virtue of an involvement in ciliary recycling. Previously, we reported that Trypanosoma brucei Rab23 localized to the nuclear envelope 1, which is clearly inconsistent with the mammalian location and function. As T. brucei is unicellular the potential that Rab23 has no role in cell signaling was possible. Here we sought to further investigate the role(s of Rab23 in T. brucei to determine if Rab23 was an example of a Rab protein with divergent function in distinct taxa. Methods/major findings The taxonomic distribution of Rab23 was examined and compared with the presence of flagella/cilia in representative taxa. Despite evidence for considerable secondary loss, we found a clear correlation between a conventional flagellar structure and the presence of a Rab23 orthologue in the genome. By epitope-tagging, Rab23 was localized and found to be present at the flagellum throughout the cell cycle. However, RNAi knockdown did not result in a flagellar defect, suggesting that Rab23 is not required for construction or maintenance of the flagellum. Conclusions The location of Rab23 at the flagellum is conserved between mammals and trypanosomes and the Rab23 gene is restricted to flagellated organisms. These data may suggest the presence of a Rab23-mediated signaling mechanism in trypanosomes.

  14. Colon cancer screening

    Science.gov (United States)

    Screening for colon cancer; Colonoscopy - screening; Sigmoidoscopy - screening; Virtual colonoscopy - screening; Fecal immunochemical test; Stool DNA test; sDNA test; Colorectal cancer - screening; Rectal ...

  15. Comprehensive proteomic analysis of Trypanosoma cruzi epimastigote cell surface proteins by two complementary methods

    DEFF Research Database (Denmark)

    Queiroz, Rayner M L; Charneau, Sébastien; Motta, Flávia N

    2013-01-01

    Trypanosoma cruzi is a protozoan that causes Chagas' disease, a neglected infectious illness that affects millions of people, mostly in Latin America. Here, the cell surface subproteome of the T. cruzi epimastigote life form was characterized. In order to prepare samples enriched in epimastigote...

  16. Cell surface proteome analysis of human-hosted Trypanosoma cruzi life stages

    DEFF Research Database (Denmark)

    Queiroz, Rayner M L; Charneau, Sébastien; Bastos, Izabela M D

    2014-01-01

    Chagas' disease is a neglected infectious illness, caused by the protozoan Trypanosoma cruzi. It remains a challenging health issue in Latin America, where it is endemic, and so far there is no immunoprophylatic vaccine or satisfactory chemotherapic treatment for its chronic stage. The present work...

  17. Kinetic properties and inhibition of Trypanosoma cruzi 3-hydroxy-3-methylglutaryl CoA reductase

    DEFF Research Database (Denmark)

    Hurtado-Guerrrero, Ramón; Pena Diaz, Javier; Montalvetti, Andrea

    2002-01-01

    A detailed kinetic analysis of the recombinant soluble enzyme 3-hydroxy-3-methylglutaryl CoA reductase (HMGR) from Trypanosoma cruzi has been performed. The enzyme catalyzes the normal anabolic reaction and the reductant is NADPH. It also catalyzes the oxidation of mevalonate but at a lower propo...

  18. The effect of the diterpene 5-epi-icetexone on the cell cycle of Trypanosoma cruzi.

    NARCIS (Netherlands)

    Lozano, E.; Barrera, P.; Tonn, C.; Nieto, M.; Sartor, T.; Sosa, M.A.

    2012-01-01

    Numerous natural compounds have been used against Trypanosoma cruzi, the causative agent of Chagas' disease. Here, we studied the effect of the diterpene 5-epi-icetexone on growth and morphology of parasites synchronized with hydroxyurea, at different periods of time after removal of the nucleotide.

  19. Lack of evidence for integration of Trypanosoma cruzi minicircle DNA in South American human genomes

    Czech Academy of Sciences Publication Activity Database

    Flegontova, Olga; Lukeš, Julius; Flegontov, Pavel

    2012-01-01

    Roč. 42, č. 5 (2012), s. 437-441 ISSN 0020-7519 Grant - others:GA MŠk(CZ) LM2010005 Institutional support: RVO:60077344 Keywords : Trypanosoma cruzi * Kinetoplast minicircle * Chagas disease * Horizontal gene transfer * Human genome Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.637, year: 2012 http://www.sciencedirect.com/science/article/pii/S0020751912000781

  20. Analysis of the mitochondrial maxicircle of Trypanosoma lewisi, a neglected human pathogen

    Czech Academy of Sciences Publication Activity Database

    Lin, R.-H.; Lai, D.-H.; Zheng, L.-L.; Wu, J.; Lukeš, Julius; Hide, G.; Lun, Z.-R.

    2015-01-01

    Roč. 8, 30 December 2015 (2015), s. 665 ISSN 1756-3305 Institutional support: RVO:60077344 Keywords : Trypanosoma lewisi * Kinetoplast maxicircle * Mitochondrial DNA * RNA editing * Palindrome Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.234, year: 2015

  1. YCF45 protein, usually associated with plastids, is targeted into the mitochondrion of Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Týč, Jiří; Long, Shaojun; Jirků, Milan; Lukeš, Julius

    2010-01-01

    Roč. 173, č. 1 (2010), s. 43-47 ISSN 0166-6851 R&D Projects: GA ČR GA204/09/1667 Institutional research plan: CEZ:AV0Z60220518 Keywords : Trypanosoma * Plastid * Mitochondrion * Targeting * YCF45 * Horizontal gene transfer Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.875, year: 2010

  2. AcSDKP is down-regulated in anaemia induced by Trypanosoma ...

    African Journals Online (AJOL)

    We studied the responses of a tetrapeptide, AcSDKP, and IL-10, and their association with bone marrow nucleated cells in a Trypanosoma brucei brucei GVR35 experimental infection model. Methods Mouse infection was done intraperitoneally with 1 × 103 trypanosomes/mL. Mice were either infected or left uninfected (N ...

  3. Efficacy of some essential oils in mice infected with Trypanosoma cruzi

    African Journals Online (AJOL)

    Purpose: To evaluate the efficacy of orally administered Cymbopogon citratus, Zingiber officinale and Syzygium aromaticum essential oils (EOs) in mice infected with Trypanosoma cruzi. Methods: Three experiments were conducted with 48 Swiss mice each. The animals were inoculated with 2 x 106 metacyclic ...

  4. Molecular variation of Trypanosoma brucei subspecies as revealed by AFLP fingerprinting

    NARCIS (Netherlands)

    Agbo, E.E.C.; Majiwa, P.A.O.; Claassen, H.J.H.M.; Pas, te M.F.W.

    2002-01-01

    Genetic analysis of Trypanosoma spp. depends on the detection of variation between strains. We have used the amplified fragment length polymorphism (AFLP) technique to develop a convenient and reliable method for genetic characterization of Trypanosome (sub)species. AFLP accesses multiple

  5. Futile import of tRNAs and proteins into the mitochondrion of Trypanosoma brucei evansi

    Czech Academy of Sciences Publication Activity Database

    Paris, Zdeněk; Hashimi, Hassan; Lun, Sijia; Alfonzo, J. D.; Lukeš, Julius

    2011-01-01

    Roč. 176, č. 2 (2011), 116-120 ISSN 0166-6851 R&D Projects: GA ČR GA204/09/1667; GA MŠk LC07032 Institutional research plan: CEZ:AV0Z60220518 Keywords : Trypanosoma * tRNA * Protein import * Mitochondrion * Kinetoplast Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.551, year: 2011

  6. Functions and cellular localization of cysteine desulfurase and selenocysteine lyase in Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Poliak, Pavel; Van Hoewyk, D.; Oborník, Miroslav; Zíková, Alena; Stuart, K. D.; Tachezy, J.; Pilon, M.; Lukeš, Julius

    2010-01-01

    Roč. 277, č. 2 (2010), s. 383-393 ISSN 1742-464X R&D Projects: GA ČR GA204/09/1667 Institutional research plan: CEZ:AV0Z60220518 Keywords : Fe–S cluster * mitochondrion * RNAi * selenoprotein * Trypanosoma Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.129, year: 2010

  7. The promoter for a variant surface glycoprotein gene expression site in Trypanosoma brucei

    NARCIS (Netherlands)

    Zomerdijk, J. C.; Ouellette, M.; ten Asbroek, A. L.; Kieft, R.; Bommer, A. M.; Clayton, C. E.; Borst, P.

    1990-01-01

    The variant-specific surface glycoprotein (VSG) gene 221 of Trypanosoma brucei is transcribed as part of a 60 kb expression site (ES). We have identified the promoter controlling this multigene transcription unit by the use of 221 chromosome-enriched DNA libraries and VSG gene 221 expression site

  8. Molecular Confirmation of Trypanosoma evansi and Babesia bigemina in Cattle from Lower Egypt

    Directory of Open Access Journals (Sweden)

    Mahmoud M. Elhaig, Abdelfattah Selim, Mohamed M. Mahmoud and Eman K El-Gayar

    2016-11-01

    Full Text Available Trypanosomosis and babesiosis are economically important vector-borne diseases for animal health and productivity in developing countries. In Egypt, molecular epidemiological surveys on such diseases are scarce. In the present study, we examined 475 healthy and 25 clinically diagnosed cattle from three provinces in Lower Egypt, for Trypanosoma (T. and Babesia (B. infections using an ITS1 PCR assay that confirmed Trypanosoma species presence and an 18S rRNA assay that detected B. bigemina. Results confirmed Trypanosoma spp. and B. bigemina presence in 30.4% and 11% individuals, respectively, with eight animals (1.6% being co-infected with both hemoparasites. Subsequent type-specific PCRs revealed that all Trypanosoma PCR positive samples corresponded to T. evansi and that none of the animals harboured T. brucei gambiense or T. brucei rhodesiense. Nucleotide sequencing of the variable surface glycoprotein revealed the T. evansi cattle strain to be most closely related (99% nucleotide sequence identity to strains previously detected in dromedary camels in Egypt, while the 18S rRNA gene phylogeny confirmed the presence of a unique B. bigemina haplotype closely related to strains from Turkey and Brazil. Statistically significant differences in PCR prevalence were noted with respect to gender, clinical status and locality. These results confirm the presence of high numbers of carrier animals and signal the need for expanded surveillance and control efforts.

  9. Non-cytochrome mediated mitochondrial ATP production in bloodstream form Trypanosoma brucei brucei

    NARCIS (Netherlands)

    Bienen, E. J.; Maturi, R. K.; Pollakis, G.; Clarkson, A. B.

    1993-01-01

    The life cycle of Trypanosoma brucei brucei involves a series of differentiation steps characterized by marked changes in mitochondrial development and function. The bloodstream forms of this parasite completely lack cytochromes and have not been considered to have any Krebs cycle function. It has

  10. Procyclic Trypanosoma brucei do not use Krebs cycle activity for energy generation

    NARCIS (Netherlands)

    Weelden, van S.W.H.; Fast, B.; Vogt, A.; Meer, van der P.; Saas, J.; Hellemond, van J.J.; Tielens, A.G.M.; Boshart, M.

    2003-01-01

    The importance of a functional Krebs cycle for energy generation in the procyclic stage of Trypanosoma brucei was investigated under physiological conditions during logarithmic phase growth of a pleomorphic parasite strain. Wild type procyclic cells and mutants with targeted deletion of the gene

  11. Trypanocidal action of bisphosphonium salts through a mitochondrial target in bloodstream form Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Alkhaldi, A.A.M.; Martínek, Jan; Panicucci, Brian; Dardonville, C.; Zíková, Alena; de Koning, H.P.

    2016-01-01

    Roč. 6, č. 1 (2016), s. 23-34 ISSN 2211-3207 R&D Projects: GA MŠk LL1205 Institutional support: RVO:60077344 Keywords : Trypanosoma brucei * mitochondrion * FoF1 ATPase * succinate dehydrogenase * phosphonium salt * SDH complex Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.809, year: 2016

  12. Trypanosoma cruzi in the chicken model: Chagas-like heart disease in the absence of parasitism

    Czech Academy of Sciences Publication Activity Database

    Teixeira, A.R.L.; Gomes, C.; Nitz, N.; Sousa, A.O.; Alvez, R.M.; Guimaro, M.C.; Cordeiro, C.; Bernal, F.M.; Rosa, A.C.; Hejnar, Jiří; Leonardecz, E.; Hecht, M.M.

    2011-01-01

    Roč. 5, č. 3 (2011), e1000 ISSN 1935-2735 Institutional research plan: CEZ:AV0Z50520514 Keywords : Chagas disease * Trypanosoma cruzi * kDNA minicircles * inbred chicken Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.716, year: 2011

  13. Extraction of Trypanosoma cruzi DNA from food: a contribution to the elucidation of acute Chagas disease outbreaks.

    Science.gov (United States)

    Ferreira, Renata Trotta Barroso; Melandre, Aline Martins; Cabral, Maria Luiza; Branquinho, Maria Regina; Cardarelli-Leite, Paola

    2016-04-01

    Before 2004, the occurrence of acute Chagas disease (ACD) by oral transmission associated with food was scarcely known or investigated. Originally sporadic and circumstantial, ACD occurrences have now become frequent in the Amazon region, with recently related outbreaks spreading to several Brazilian states. These cases are associated with the consumption of açai juice by waste reservoir animals or insect vectors infected with Trypanosoma cruzi in endemic areas. Although guidelines for processing the fruit to minimize contamination through microorganisms and parasites exist, açai-based products must be assessed for quality, for which the demand for appropriate methodologies must be met. Dilutions ranging from 5 to 1,000 T. cruzi CL Brener cells were mixed with 2mL of acai juice. Four Extraction of T. cruzi DNA methods were used on the fruit, and the cetyltrimethyl ammonium bromide (CTAB) method was selected according to JRC, 2005. DNA extraction by the CTAB method yielded satisfactory results with regard to purity and concentration for use in PCR. Overall, the methods employed proved that not only extraction efficiency but also high sensitivity in amplification was important. The method for T. cruzi detection in food is a powerful tool in the epidemiological investigation of outbreaks as it turns epidemiological evidence into supporting data that serve to confirm T. cruzi infection in the foods. It also facilitates food quality control and assessment of good manufacturing practices involving acai-based products.

  14. Extraction of Trypanosoma cruzi DNA from food: a contribution to the elucidation of acute Chagas disease outbreaks

    Directory of Open Access Journals (Sweden)

    Renata Trotta Barroso Ferreira

    2016-04-01

    Full Text Available Abstract: INTRODUCTION: Before 2004, the occurrence of acute Chagas disease (ACD by oral transmission associated with food was scarcely known or investigated. Originally sporadic and circumstantial, ACD occurrences have now become frequent in the Amazon region, with recently related outbreaks spreading to several Brazilian states. These cases are associated with the consumption of açai juice by waste reservoir animals or insect vectors infected with Trypanosoma cruzi in endemic areas. Although guidelines for processing the fruit to minimize contamination through microorganisms and parasites exist, açai-based products must be assessed for quality, for which the demand for appropriate methodologies must be met. METHODS: Dilutions ranging from 5 to 1,000 T. cruzi CL Brener cells were mixed with 2mL of acai juice. Four Extraction of T. cruzi DNA methods were used on the fruit, and the cetyltrimethyl ammonium bromide (CTAB method was selected according to JRC, 2005. RESULTS: DNA extraction by the CTAB method yielded satisfactory results with regard to purity and concentration for use in PCR. Overall, the methods employed proved that not only extraction efficiency but also high sensitivity in amplification was important. CONCLUSIONS: The method for T. cruzi detection in food is a powerful tool in the epidemiological investigation of outbreaks as it turns epidemiological evidence into supporting data that serve to confirm T. cruzi infection in the foods. It also facilitates food quality control and assessment of good manufacturing practices involving acai-based products.

  15. Trypanocidal activity of human plasma on Trypanosoma evansi in mice Atividade tripanocida do plasma humano sobre Trypanosoma evansi em camundongos

    Directory of Open Access Journals (Sweden)

    Aleksandro Schafer Da Silva

    2012-03-01

    Full Text Available This study aimed to test an alternative protocol with human plasma to control Trypanosoma evansi infection in mice. Plasma from an apparently 27-year-old healthy male, blood type A+, was used in the study. A concentration of 100 mg.dL-1 apolipoprotein L1 (APOL1 was detected in the plasma. Forty mice were divided into four groups with 10 animals each. Group A comprised uninfected animals. Mice from groups B, C and D were inoculated with a T. evansi isolate. Group B was used as a positive control. At three days post-infection (DPI, the mice were administered intraperitoneally with human plasma. A single dose of 0.2 mL plasma was given to those in group C. The mice from group D were administered five doses of 0.2 mL plasma with a 24 hours interval between the doses. Group B showed high increasing parasitemia that led to their death within 5 DPI. Both treatments eliminated parasites from the blood and increased the longevity of animals. An efficacy of 50 (group C and 80% (group D of human plasma trypanocidal activity was found using PCR. This therapeutic success was likely achieved in the group D due to their higher levels of APOL1 compared with group C.Este estudo teve como objetivo testar um protocolo alternativo com plasma humano para controlar a infecção por Trypanosoma evansi em camundongos. O plasma foi oriundo de um homem aparentemente saudável, com idade entre 27 anos e tipo de sangue A+. Foi detectada uma concentração de 100 mg.dL -1 de apolipoproteína L1 (APOL1 no plasma. Quarenta camundongos foram divididos em quatro grupos, contendo dez animais cada. Grupo A, composto de animais não infectados. Os roedores dos grupos B, C e D foram inoculados intraperitonealmente com um isolado de T. evansi. O Grupo B foi usado como um controle positivo. Três dias pós-infecção (DPI, os camundongos foram tratados com plasma humano. Uma dose única de 0,2 mL de plasma foi administrada nos roedores do grupo C. Os ratos do grupo D receberam cinco

  16. EXPERIMENTAL INFECTION BY Trypanosoma vivax IN GOATS INFECÇÃO EXPERIMENTAL EM CAPRINOS COM Trypanosoma vivax

    Directory of Open Access Journals (Sweden)

    Francisco David Nascimento Sousa

    2008-10-01

    Full Text Available

    Four goats were infected intravenously with 1.0 mL of cattle blood containing about 1.25 x 105 Trypanosoma vivax derived from spontaneous outbreak in cattle at Catolé do Rocha city, Paraíba, Brazil. Other four goats were used as controls. Parasitemia and body temperature were determined daily for 40 days. Animals were weighted each 7 days, and blood samples for blood cells counts were collected each 5 days. It was obtained a sample of liquor from each animal before death; cerebrospinal fluid samples were submitted to biochemical and cytological evaluations, density determination and parasite detection. A positive correlation was found between body temperature and parasitemia in infected animals. These animals presented anemia, leukopenia, hypoglycemia, decreased serum levels of total proteins and cholesterol, and nervous symptoms. Examination of cerebrospinal fluid resulted in decrease of glucose levels and increase in lactate dehydrogenase, cell counts and presence of the parasite. At necropsy it was found pale carcass, generalized infartation of lymphonodes, pulmonary edema, and liquid accumulation of pericardium. Histological changes were characterized by interstitial pneumonia, miocarditis, cardiac fibrosis, meningitis, and encephalitis. All observed changes confirm patogenicity of T. vivax.

    KEY WORDS: Experimental infection, trypanosomiasis, patogenicity.

    Quatro caprinos foram infectados experimentalmente por via intravenosa com 1,0 ml de sangue contendo aproximadamente 1,25 x 105 tripanossomas/ml, utilizando-se um isolado de Trypanosoma vivax de bovinos infectados naturalmente no município de Catolé do Rocha, Paraíba. A parasitemia e a temperatura foram determinadas diariamente durante quarenta dias. A cada cinco dias realizaram-se coletas de sangue para hemograma e análise bioquímica sérica. Antes do

  17. Automated high-content assay for compounds selectively toxic to Trypanosoma cruzi in a myoblastic cell line.

    Directory of Open Access Journals (Sweden)

    Julio Alonso-Padilla

    2015-01-01

    Full Text Available Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, represents a very important public health problem in Latin America where it is endemic. Although mostly asymptomatic at its initial stage, after the disease becomes chronic, about a third of the infected patients progress to a potentially fatal outcome due to severe damage of heart and gut tissues. There is an urgent need for new drugs against Chagas disease since there are only two drugs available, benznidazole and nifurtimox, and both show toxic side effects and variable efficacy against the chronic stage of the disease.Genetically engineered parasitic strains are used for high throughput screening (HTS of large chemical collections in the search for new anti-parasitic compounds. These assays, although successful, are limited to reporter transgenic parasites and do not cover the wide T. cruzi genetic background. With the aim to contribute to the early drug discovery process against Chagas disease we have developed an automated image-based 384-well plate HTS assay for T. cruzi amastigote replication in a rat myoblast host cell line. An image analysis script was designed to inform on three outputs: total number of host cells, ratio of T. cruzi amastigotes per cell and percentage of infected cells, which respectively provides one host cell toxicity and two T. cruzi toxicity readouts. The assay was statistically robust (Z´ values >0.6 and was validated against a series of known anti-trypanosomatid drugs.We have established a highly reproducible, high content HTS assay for screening of chemical compounds against T. cruzi infection of myoblasts that is amenable for use with any T. cruzi strain capable of in vitro infection. Our visual assay informs on both anti-parasitic and host cell toxicity readouts in a single experiment, allowing the direct identification of compounds selectively targeted to the parasite.

  18. Characterization and immobilization of engineered sialidases from Trypanosoma rangeli for transsialylation

    Directory of Open Access Journals (Sweden)

    Birgitte Zeuner

    2017-04-01

    Full Text Available A sialidase (EC 3.2.1.18; GH 33 from non-pathogenic Trypanosoma rangeli has been engineered with the aim of improving its transsialylation activity. Recently, two engineered variants containing 15 and 16 amino acid substitutions, respectively, were found to exhibit significantly improved transsialylation activity: both had a 14 times higher ratio between transsialylation and hydrolysis products compared to the first reported mutant TrSA5mut. In the current work, these two variants, Tr15 and Tr16, were characterized in terms of pH optimum, thermal stability, effect of acceptor-to-donor ratio, and acceptor specificity for transsialylation using casein glycomacropeptide (CGMP as sialyl donor and lactose or other human milk oligosaccharide core structures as acceptors. Both sialidase variants exhibited pH optima around pH 4.8. Thermal stability of each enzyme was comparable to that of previously developed T. rangeli sialidase variants and higher than that of the native transsialidase from T. cruzi (TcTS. As for other engineered T. rangeli sialidase variants and TcTS, the acceptor specificity was broad: lactose, galactooligosaccharides (GOS, xylooligosaccharides (XOS, and human milk oligosaccharide structures lacto-N-tetraose (LNT, lacto-N-fucopentaose (LNFP V, and lacto-N-neofucopentaose V (LNnFP V were all sialylated by Tr15 and Tr16. An increase in acceptor-to-donor ratio from 2 to 10 had a positive effect on transsialylation. Both enzymes showed high preference for formation α(2,3-linkages at the non-reducing end of lactose in the transsialylation. Tr15 was the most efficient enzyme in terms of transsialylation reaction rates and yield of 3’-sialyllactose. Finally, Tr15 was immobilized covalently on glyoxyl-functionalized silica, leading to a 1.5-fold increase in biocatalytic productivity (mg 3’-sialyllactose per mg enzyme compared to free enzyme after 6 cycles of reuse. The use of glyoxyl-functionalized silica proved to be markedly better

  19. Mating compatibility in the parasitic protist Trypanosoma brucei.

    Science.gov (United States)

    Peacock, Lori; Ferris, Vanessa; Bailey, Mick; Gibson, Wendy

    2014-02-21

    Genetic exchange has been described in several kinetoplastid parasites, but the most well-studied mating system is that of Trypanosoma brucei, the causative organism of African sleeping sickness. Sexual reproduction takes place in the salivary glands (SG) of the tsetse vector and involves meiosis and production of haploid gametes. Few genetic crosses have been carried out to date and consequently there is little information about the mating compatibility of different trypanosomes. In other single-celled eukaryotes, mating compatibility is typically determined by a system of two or more mating types (MT). Here we investigated the MT system in T. brucei. We analysed a large series of F1, F2 and back crosses by pairwise co-transmission of red and green fluorescent cloned cell lines through experimental tsetse flies. To analyse each cross, trypanosomes were cloned from fly SG containing a mixture of both parents, and genotyped by microsatellites and molecular karyotype. To investigate mating compatibility at the level of individual cells, we directly observed the behaviour of SG-derived gametes in intra- or interclonal mixtures of red and green fluorescent trypanosomes ex vivo. Hybrid progeny were found in all F1 and F2 crosses and most of the back crosses. The success of individual crosses was highly variable as judged by the number of hybrid clones produced, suggesting a range of mating compatibilities among F1 progeny. As well as hybrids, large numbers of recombinant genotypes resulting from intraclonal mating (selfers) were found in some crosses. In ex vivo mixtures, red and green fluorescent trypanosome gametes were observed to pair up and interact via their flagella in both inter- and intraclonal combinations. While yellow hybrid trypanosomes were frequently observed in interclonal mixtures, such evidence of cytoplasmic exchange was rare in the intraclonal mixtures. The outcomes of individual crosses, particularly back crosses, were variable in numbers of both

  20. Intraclonal mating occurs during tsetse transmission of Trypanosoma brucei

    Directory of Open Access Journals (Sweden)

    Ferris Vanessa

    2009-09-01

    Full Text Available Abstract Background Mating in Trypanosoma brucei is a non-obligatory event, triggered by the co-occurrence of different strains in the salivary glands of the vector. Recombinants that result from intra- rather than interclonal mating have been detected, but only in crosses of two different trypanosome strains. This has led to the hypothesis that when trypanosomes recognize a different strain, they release a diffusible factor or pheromone that triggers mating in any cell in the vicinity whether it is of the same or a different strain. This idea assumes that the trypanosome can recognize self and non-self, although there is as yet no evidence for the existence of mating types in T. brucei. Results We investigated intraclonal mating in T. b. brucei by crossing red and green fluorescent lines of a single strain, so that recombinant progeny can be detected in the fly by yellow fluorescence. For strain 1738, seven flies had both red and green trypanosomes in the salivary glands and, in three, yellow trypanosomes were also observed, although they could not be recovered for subsequent analysis. Nonetheless, both red and non-fluorescent clones from these flies had recombinant genotypes as judged by microsatellite and karyotype analyses, and some also had raised DNA contents, suggesting recombination or genome duplication. Strain J10 produced similar results indicative of intraclonal mating. In contrast, trypanosome clones recovered from other flies showed that genotypes can be transmitted with fidelity. When a yellow hybrid clone expressing both red and green fluorescent protein genes was transmitted, the salivary glands contained a mixture of fluorescent-coloured trypanosomes, but only yellow and red clones were recovered. While loss of the GFP gene in the red clones could have resulted from gene conversion, some of these clones showed loss of heterozygosity and raised DNA contents as in the other single strain transmissions. Our observations suggest

  1. The Complement System: A Prey of Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Kárita C. F. Lidani

    2017-04-01

    Full Text Available Trypanosoma cruzi is a protozoan parasite known to cause Chagas disease (CD, a neglected sickness that affects around 6–8 million people worldwide. Originally, CD was mainly found in Latin America but more recently, it has been spread to countries in North America, Asia, and Europe due the international migration from endemic areas. Thus, at present CD represents an important concern of global public health. Most of individuals that are infected by T. cruzi may remain in asymptomatic form all lifelong, but up to 40% of them will develop cardiomyopathy, digestive mega syndromes, or both. The interaction between the T. cruzi infective forms and host-related immune factors represents a key point for a better understanding of the physiopathology of CD. In this context, the complement, as one of the first line of host defense against infection was shown to play an important role in recognizing T. cruzi metacyclic trypomastigotes and in controlling parasite invasion. The complement consists of at least 35 or more plasma proteins and cell surface receptors/regulators, which can be activated by three pathways: classical (CP, lectin (LP, and alternative (AP. The CP and LP are mainly initiated by immune complexes or pathogen-associated molecular patterns (PAMPs, respectively, whereas AP is spontaneously activated by hydrolysis of C3. Once activated, several relevant complement functions are generated which include opsonization and phagocytosis of particles or microorganisms and cell lysis. An important step during T. cruzi infection is when intracellular trypomastigotes are release to bloodstream where they may be target by complement. Nevertheless, the parasite uses a sequence of events in order to escape from complement-mediated lysis. In fact, several T. cruzi molecules are known to interfere in the initiation of all three pathways and in the assembly of C3 convertase, a key step in the activation of complement. Moreover, T. cruzi promotes secretion

  2. Aspects of resistance to experimental infection with Trypanosoma cruzi

    International Nuclear Information System (INIS)

    Dias, Viviane Liotti

    2010-01-01

    Chagas disease, a zoonosis caused by the protozoan Trypanosoma cruzi, has a wide distribution in Latin America and extends from the southern part of the United States to Argentina. A number of 10 million of infected people is estimated and another 25 million exposed to the risk. Although discovered over a century, Chagas disease is still a serious infection that causes great socioeconomic impact, with no effective treatment at the chronic phase and in which, a lack of scientific knowledge can be observed. The main goal of this work was that obtaining and using consomic strain of mice, the resistance could be investigated. Consomic strains were produced by programmed mating, in which the animals were monitored with DNA polymorphic markers, and one of his chromosomes was replaced by his homologue from another strain. As parental, were used, the inbred strains C57BL/6/J Unib with resistant phenotype (donor) and as receiver, the A/JUnib strain, that has a susceptible phenotype. These models were used to produce five consomic strains: for the chromosomes 7 (CSs7), 11 (CSs11), 14 (CSs14), 17 (CSs17) and 19 (CSs19), described by Passos et al. (2003) as important in controlling infection caused by the Y strain of T. cruzi. In experimental testing, the consomics were inoculated intraperitoneally at doses of 10 1 , 10 2 , 10 3 and 10 4 using as control, animals from both parental lines. In all consomics, resistance was higher than that observed in the susceptible parental. In a second protocol, the consomics were mated with scheduled associations and the progenies were challenged with inocula employing increasing doses of trypomastigotes. The resistance observed in this group was also higher than that observed in the parental with susceptible phenotype. The observed results demonstrate that the use of the consomic strains that were produced order to assess the contribution of each chromosome in the resistance, as well as the effects of association between chromosomes are an

  3. Zoonotic trypanosomes in South East Asia : attempts to control Trypanosoma lewisi using human and animal trypanocidal drugs

    OpenAIRE

    Desquesnes, M.; Yangtara, S.; Kunphukhieo, P.; Jittapalapong, S.; Herder, Stéphane

    2016-01-01

    Beside typical human trypanosomes responsible of sleeping sickness in Africa and Chagas disease in Latin America, there is a growing number of reported atypical human infections due to Trypanosoma evansi, a livestock parasite, or Trypanosoma lewisi, a rat parasite, especially in Asia. Drugs available for the treatment of T. brucei ssp. in humans are obviously of choice for the control of T. evansi because it is derived from T. brucei. However, concerning T. lewisi, there is an urgent need to ...

  4. Soviet test yields

    Science.gov (United States)

    Vergino, Eileen S.

    Soviet seismologists have published descriptions of 96 nuclear explosions conducted from 1961 through 1972 at the Semipalatinsk test site, in Kazakhstan, central Asia [Bocharov et al., 1989]. With the exception of releasing news about some of their peaceful nuclear explosions (PNEs) the Soviets have never before published such a body of information.To estimate the seismic yield of a nuclear explosion it is necessary to obtain a calibrated magnitude-yield relationship based on events with known yields and with a consistent set of seismic magnitudes. U.S. estimation of Soviet test yields has been done through application of relationships to the Soviet sites based on the U.S. experience at the Nevada Test Site (NTS), making some correction for differences due to attenuation and near-source coupling of seismic waves.

  5. Aspects of resistance to experimental infection with Trypanosoma cruzi; Aspectos da resistencia a infecao experimental com Trypanosoma cruzi

    Energy Technology Data Exchange (ETDEWEB)

    Dias, Viviane Liotti

    2010-07-01

    Chagas disease, a zoonosis caused by the protozoan Trypanosoma cruzi, has a wide distribution in Latin America and extends from the southern part of the United States to Argentina. A number of 10 million of infected people is estimated and another 25 million exposed to the risk. Although discovered over a century, Chagas disease is still a serious infection that causes great socioeconomic impact, with no effective treatment at the chronic phase and in which, a lack of scientific knowledge can be observed. The main goal of this work was that obtaining and using consomic strain of mice, the resistance could be investigated. Consomic strains were produced by programmed mating, in which the animals were monitored with DNA polymorphic markers, and one of his chromosomes was replaced by his homologue from another strain. As parental, were used, the inbred strains C57BL/6/J Unib with resistant phenotype (donor) and as receiver, the A/JUnib strain, that has a susceptible phenotype. These models were used to produce five consomic strains: for the chromosomes 7 (CSs7), 11 (CSs11), 14 (CSs14), 17 (CSs17) and 19 (CSs19), described by Passos et al. (2003) as important in controlling infection caused by the Y strain of T. cruzi. In experimental testing, the consomics were inoculated intraperitoneally at doses of 10{sup 1}, 10{sup 2}, 10{sup 3} and 10{sup 4} using as control, animals from both parental lines. In all consomics, resistance was higher than that observed in the susceptible parental. In a second protocol, the consomics were mated with scheduled associations and the progenies were challenged with inocula employing increasing doses of trypomastigotes. The resistance observed in this group was also higher than that observed in the parental with susceptible phenotype. The observed results demonstrate that the use of the consomic strains that were produced order to assess the contribution of each chromosome in the resistance, as well as the effects of association between

  6. Screen dealing

    International Nuclear Information System (INIS)

    Barlow, J.W.

    1991-01-01

    The screen dealing system provides a facility whereby buyers and sellers of spot thermal coal can make bids and offers via the medium of the Reuters screen. A sale results when a market participant notifies his acceptance of a price to a central dealing desk. Use of the system is available to all genuine participants in the coal trade. This paper reports that it provides a focus for information and for the visible making of coal prices. For years screen trading has been used successfully to trade other commodities. At last coal is being traded electronically. It makes sense. It works. Users like it

  7. Enhancement of HHG yield

    International Nuclear Information System (INIS)

    Serrat, C.; Biegert, J.

    2011-01-01

    A static electric field periodically distributed in space controls and enhances the yield in high harmonic generation. The method is relatively simple to implement and allows tuning from the extreme-ultraviolet to soft X-ray. The radiation yield is selectively enhanced due to symmetry breaking induced by a static electric field on the interaction between the driving laser and the medium. The enhanced spectral region is tuned by varying the periodicity of the static electric field. Simulations predict an increase of more than two orders of magnitude for harmonics in the water window spectral range.

  8. Airport Screening

    Science.gov (United States)

    Health Physics Society Specialists in Radiation Safety Airport Screening Fact Sheet Adopted: May 2011 Photo courtesy of Dan ... a safe level. An American National Standards Institute/Health Physics Society industry standard states that the maxi- mum ...

  9. Hypertension screening

    Science.gov (United States)

    Foulke, J. M.

    1975-01-01

    An attempt was made to measure the response to an announcement of hypertension screening at the Goddard Space Center, to compare the results to those of previous statistics. Education and patient awareness of the problem were stressed.

  10. Carrier Screening

    Science.gov (United States)

    ... How accurate is carrier screening? No test is perfect. In a small number of cases, test results ... in which an egg is removed from a woman’s ovary, fertilized in a laboratory with the man’s ...

  11. Molecular characterization and interactome analysis of Trypanosoma cruzi tryparedoxin II.

    Science.gov (United States)

    Arias, Diego G; Piñeyro, María Dolores; Iglesias, Alberto A; Guerrero, Sergio A; Robello, Carlos

    2015-04-29

    Trypanosoma cruzi, the causative agent of Chagas disease, possesses two tryparedoxins (TcTXNI and TcTXNII), belonging to the thioredoxin superfamily. TXNs are oxidoreductases which mediate electron transfer between trypanothione and peroxiredoxins. This constitutes a difference with the host cells, in which these activities are mediated by thioredoxins. These differences make TXNs an attractive target for drug development. In a previous work we characterized TcTXNI, including the redox interactome. In this work we extend the study to TcTXNII. We demonstrate that TcTXNII is a transmembrane protein anchored to the surface of the mitochondria and endoplasmic reticulum, with a cytoplasmatic orientation of the redox domain. It would be expressed during the metacyclogenesis process. In order to continue with the characterization of the redox interactome of T. cruzi, we designed an active site mutant TcTXNII lacking the resolving cysteine, and through the expression of this mutant protein and incubation with T. cruzi proteins, heterodisulfide complexes were isolated by affinity chromatography and identified by mass spectrometry. This allowed us to identify sixteen TcTXNII interacting proteins, which are involved in a wide range of cellular processes, indicating the relevance of TcTXNII, and contributing to our understanding of the redox interactome of T. cruzi. T. cruzi, the causative agent of Chagas disease, constitutes a major sanitary problem in Latin America. The number of estimated infected persons is ca. 8 million, 28 million people are at risk of infection and ~20,000 deaths occur per year in endemic regions. No vaccines are available at present, and most drugs currently in use were developed decades ago and show variable efficacy with undesirable side effects. The parasite is able to live and prolipherate inside macrophage phagosomes, where it is exposed to cytotoxic reactive oxygen and nitrogen species, derived from macrophage activation. Therefore, T. cruzi

  12. Characterization of plasma menbrane polypeptides of trypanosoma from bats Caracterização de polipeptídeos de membrana plasmática de tripanosomas de morcegos

    OpenAIRE

    R. T. Pinho; Giovanni de Simone

    1989-01-01

    Cell surface proteins of Trypanosoma dionisii, Trypanosoma vespertilionis and Trypanosoma sp. (M238) were radiodinated and their distribution both in the detergent-poor (DPP) and dertergent-enriched phase (DRP) was studied using a phase separation technique in Triton X-114 as well as polyacrylamide gel electrophoresis in sodium dodecyl sulphate (SDS-PAGE). Significant differences were observed in the proteins present in the DRP when the three species of trypanosoma were compared. Two major ba...

  13. Comprehensive glycoprofiling of the epimastigote and trypomastigote stages of Trypanosoma cruzi

    DEFF Research Database (Denmark)

    Alves, Maria Julia Manso; Kawahara, Rebeca; Viner, Rosa

    2017-01-01

    Trypanosoma cruzi, the protozoan that causes Chagas disease, has a complex life cycle involving insect and mammalian hosts and distinct developmental stages. During T. cruzi developmental stages, glycoproteins play important role in the host-parasite interaction, such as cellular recognition, host...... the significant T. cruzi stage-specific expression of glycoproteins that can help to better understand the T. cruzi phenotype and response caused by the interaction with different hosts during its complex life cycle. BIOLOGICAL SIGNIFICANCE: Chagas disease caused by the protozoan Trypanosoma cruzi is a neglected...... disease which affects millions of people especially in Latin America. The absence of efficient drugs and vaccines against Chagas disease stimulates the search for novel targets. Glycoproteins are very attractive therapeutic candidate targets since they mediate key processes in the host...

  14. A case of Trypanosoma congolense savannah type infection and its management in a dog

    Directory of Open Access Journals (Sweden)

    Peter Kimeli

    2014-12-01

    Full Text Available A case of Trypanosoma congolense savannah type infection in a 4-year old German shepherd dog weighing 26-kg was presented to the Small Animal Clinic, University of Nairobi, Kenya, with the history of anorexia and difficulty in breathing. The clinical manifestations were fever, pale mucous membrane, dyspnea and wasting. Blood examination revealed the existence of trypanosome parasites, and showed mild anemia. Internal Transcribed Spacer (ITS based polymerase chain reaction confirmed the presence of Trypanosoma congolense savannah type. Along with supporting therapy, the case was successfully managed using diminazene aceturate injection (dosed at 3.5 mg/kg body weight through intramuscular route. Complete recovery of the case was observed on day 6 of post-treatment.

  15. Anti-Trypanosoma cruzi and cytotoxic activities of Eugenia uniflora L.

    Science.gov (United States)

    Santos, Karla K A; Matias, Edinardo F F; Tintino, Saulo R; Souza, Celestina E S; Braga, Maria F B M; Guedes, Gláucia M M; Rolón, Miriam; Vega, Celeste; de Arias, Antonieta Rojas; Costa, José G M; Menezes, Irwin R A; Coutinho, Henrique D M

    2012-05-01

    Chagas disease is caused by Trypanosoma cruzi, being considered a public health problem. An alternative to combat this pathogen is the use of natural products isolated from fruits such as Eugenia uniflora, a plant used by traditional communities as food and medicine due to its antimicrobial and biological activities. Ethanolic extract from E. uniflora was used to evaluate in vitro anti-epimastigote and cytotoxic activity. This is the first record of anti-Trypanosoma activity of E. uniflora, demonstrating that a concentration presenting 50% of activity (EC(50)) was 62.76 μg/mL. Minimum inhibitory concentration (MIC) was ≤ 1024 μg/mL. Our results indicate that E. uniflora could be a source of plant-derived natural products with anti-epimastigote activity with low toxicity. Copyright © 2012 Elsevier Inc. All rights reserved.

  16. The isolation and identification of Trypanosoma cruzi from raccoons in Maryland

    Science.gov (United States)

    Walton, B.C.; Bauman, P.M.; Diamond, L.S.; Herman, C.M.

    1958-01-01

    Five raccoons trapped at Patuxent Research Refuge, Laurel, Maryland, were found to have trypanosomes in the blood which were morphologically indistinguishable from Trypanosoma cruzi on stained smears. The organism grew well in culture. It developed and reproduced in Triatoma protracta, T. infestans, T. phyllosoma, and Rhodnius prolixus. Experimental infections were produced in raccoons, opossums, mice, rats, and monkeys by inoculation of blood, culture, and triatome forms. Typical leishmaniform bodies were found in tissue sections of cardiac muscle fibers from naturally and experimentally infected animals. Cross agglutinations carried out with Iiving cultural forms and rabbit antisera demonstrated a close antigenic relationship between the raccoon trypanosome and T. cruzi (Brazil strain). On the basis of (1) morphology, (2) presence of leishmaniform tissue stages, (3) development in triatomes, (4) infectivity to a variety of mammals, (5) culture characteristics, and (6) cross reactions in serological tests, this parasite is considered conspecific with Trypanosoma cruzi (Chagas, 1909), the causative agent of American human trypanosomiasis.

  17. High yielding rice mutants for West Bengal

    International Nuclear Information System (INIS)

    Debnath, A.R.; Sen, S.

    1980-01-01

    Four high yielding mutants with specific genetic corrections of the simply inherited characters were developed from IR-8 through X-irradiation. Recurrent selections of the promising isolates were made under diverse agro-climatic conditions in Winter and Summer seasons of West Bengal. The isolates CNM 6 and CNM 25 belonging to early maturity group and CNM 20 and CNM 31, to mid-early maturity group were finally selected at X 5 generation on the basis of their resistance qualities, maturity period and grain yield. They were evaluated upto X 10 qeneration at multi-locations as Pre-release and Minikit Varieties at State level. They were also placed at the National Screening Nursery (NSN) for screening against multiple diseases and pests at the National level. CNM 6 is reported to be promising in IRTP nurseries. It is reported that CNM 25 (IET 5646) ranked 2nd on the basis of average grain yield, CNM 20 (IET 5937) and CNM 31 (IET 5936) were resistant to diseases and with yield comparable to Jaya. These four productive mutants of superior types are widely accepted. CNM 6 is recommended for cultivation in Bankura and Birbhum districts and CNM 25 and CNM 31 in the different agro-climatic zones of West Bengal. (author)

  18. Blood transfusion and iatrogenic risks in Mexico city: anti-Trypanosoma cruzi seroprevalence in 43,048 blood donors, evaluation of parasitemia, and electrocardiogram findings in seropositive

    Directory of Open Access Journals (Sweden)

    Nidia Hernández-Becerril

    2005-04-01

    Full Text Available Iatrogenous transmission of Trypanosoma cruziby blood transfusion was suggested as a potential risk by Pellegrino (1949. Seropositive blood donors in Mexico were first reported in 1978, however, limited information is available due to small sampling, the use of heterogeneous serologic assays, and geographically limited studies. A wide survey carried out in 18 out of the 32 states of Mexico, showed a national mean of 1.6% seropositive among 64,969 donors, ranging from 0.2 to 2.8%. In the present study, we have screened 43,048 voluntary blood donors in a period of five years at the Instituto Nacional de Cardiología I. Chávez, a concentration hospital located in Mexico city which serves mainly the metropolitan area and accepts from all over the country. Standardized ELISA and IIF were used to identify seropositive individuals in addition to hemoculture, PCR and standard 12 lead ECG tests that were applied to a group of seropositive patients (29/161. The result showed a seropositivity of 0.37% (161/43,048. From the group of seropositive individuals 40% (12/29 were potential carriers of T. cruzi at the donation time and 5/29 had subclinical ECG abnormalities. Parasitological tests performed in 70 erythrocyte and platelet fractions from seropositive units (70/161 showed negative results. Our findings strongly support T. cruzi screening in the transfusion medicine practice and identify subclinical heart disease among seropositive blood donors.

  19. Genetic control of resistance to Trypanosoma brucei brucei infection in mice

    Czech Academy of Sciences Publication Activity Database

    Šíma, Matyáš; Havelková, Helena; Quan, L.; Svobodová, M.; Jarošíková, T.; Vojtíšková, Jarmila; Stassen, A. P. M.; Demant, P.; Lipoldová, Marie

    2011-01-01

    Roč. 5, č. 6 (2011), e1173 ISSN 1935-2735 R&D Projects: GA AV ČR IAA500520606; GA MŠk(CZ) LC06009 Grant - others:NIH-NCI(US) 1R01CA127162-01 Institutional research plan: CEZ:AV0Z50520514 Keywords : Trypanosoma brucei brucei * mouse recombinant congenic strains * Tbbr Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.716, year: 2011

  20. Genome and transcriptome studies of the protozoan parasites Trypanosoma cruzi and Giardia intestinalis

    OpenAIRE

    Franzén, Oscar

    2012-01-01

    Trypanosoma cruzi and Giardia intestinalis are two human pathogens and protozoan parasites responsible for the diseases Chagas disease and giardiasis, respectively. Both diseases cause su ering and illness in several million individuals. The former disease occurs primarily in South America and Central America, and the latter disease occurs worldwide. Current therapeutics are toxic and lack e cacy, and potential vaccines are far from the market. Increased knowledge about the bio...

  1. Population genetic analysis of Colombian Trypanosoma cruzi isolates revealed by enzyme electrophoretic profiles

    OpenAIRE

    Ruiz-Garcia, Manuel; Montilla, Marleny; Nicholls, Sebastian; Alvarez, Diana

    2001-01-01

    Although Colombia presents an enormous biological diversity, few studies have been conducted on the population genetics of Trypanosoma cruzi. This study was carried out with 23 Colombian stocks of this protozoa analyzed for 13 isoenzymatic loci. The Hardy-Weinberg equilibrium, the genetic diversity and heterogeneity, the genetic relationships and the possible spatial structure of these 23 Colombian stocks of T. cruzi were estimated. The majority of results obtained are in agreement with a clo...

  2. First Case of Natural Infection in Pigs: Review of Trypanosoma cruzi Reservoirs in Mexico

    Directory of Open Access Journals (Sweden)

    Paz María Salazar-Schettino

    1997-07-01

    Full Text Available An epidemiological research project was performed in the State of Morelos including collection of samples for blood smears and culture, serological tests, and xenodiagnoses from a total of 76 domestic and peridomestic mammals. Two strains of Trypanosoma cruzi were isolated by haemocultures; one from a pig (Sus scrofa, the first case of natural infection reported in Mexico, and the other from a dog (Canis familiaris. This study summarizes current information in Mexico concerning confirmed reservoirs of T. cruzi

  3. The import and function of diatom and plant frataxins in the mitochondrion of Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Long, Shaojun; Vávrová, Zuzana; Lukeš, Julius

    2008-01-01

    Roč. 162, č. 1 (2008), s. 100-104 ISSN 0166-6851 R&D Projects: GA AV ČR IAA500960705; GA MŠk LC07032; GA MŠk 2B06129; GA ČR GA204/06/1558 Institutional research plan: CEZ:AV0Z60220518 Keywords : frataxin * mitochondrion * Trypanosoma * diatom * evolutionary conservativeness * import Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.951, year: 2008

  4. DEAD-box RNA helicase is dispensable for mitochondrial translation in Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Richterová, Lenka; Vávrová, Zuzana; Lukeš, Julius

    2011-01-01

    Roč. 127, č. 1 (2011), 300-303 ISSN 0014-4894 R&D Projects: GA ČR GA204/09/1667; GA MŠk LC07032; GA MŠk 2B06129 Institutional research plan: CEZ:AV0Z60220518 Keywords : Trypanosoma * Mitochondrial translation * RNA helicase * Cytochrome c oxidase * Mitochondrion Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.122, year: 2011

  5. Mitochondrial fatty acid synthesis is required for normal mitochondrial morphology and function in Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Guler, J. L.; Kriegová, Eva; Smith, T. K.; Lukeš, Julius; Englund, P. T.

    2008-01-01

    Roč. 67, č. 5 (2008), s. 1125-1142 ISSN 0950-382X R&D Projects: GA ČR GA204/06/1558; GA MŠk LC07032; GA MŠk 2B06129 Grant - others:NIH(US) AI21334; Wellcome Trust(GB) 067441 Institutional research plan: CEZ:AV0Z60220518 Keywords : Trypanosoma * mitochondrion * fatty acid * RNA interference Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.213, year: 2008

  6. Disparate phenotypic effects from the knockdown of various Trypanosoma brucei cytochrome c oxidase subunits

    Czech Academy of Sciences Publication Activity Database

    Gnipová, Anna; Panicucci, Brian; Paris, Zdeněk; Verner, Zdeněk; Horváth, A.; Lukeš, Julius; Zíková, Alena

    2012-01-01

    Roč. 184, č. 2 (2012), s. 90-98 ISSN 0166-6851 R&D Projects: GA AV ČR KJB500960901; GA ČR GA204/09/1667 Institutional research plan: CEZ:AV0Z60220518 Keywords : Trypanosoma * RNA interference * Mitochondrion * Respiratory complexes * Cytochrome c oxidase Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.734, year: 2012 http://www.sciencedirect.com/science/article/pii/S0166685112001065#

  7. The MASP family of Trypanosoma cruzi: changes in gene expression and antigenic profile during the acute phase of experimental infection.

    Directory of Open Access Journals (Sweden)

    Sara Lopes dos Santos

    Full Text Available Trypanosoma cruzi is the etiological agent of Chagas disease, a debilitating illness that affects millions of people in the Americas. A major finding of the T. cruzi genome project was the discovery of a novel multigene family composed of approximately 1,300 genes that encode mucin-associated surface proteins (MASPs. The high level of polymorphism of the MASP family associated with its localization at the surface of infective forms of the parasite suggests that MASP participates in host-parasite interactions. We speculate that the large repertoire of MASP sequences may contribute to the ability of T. cruzi to infect several host cell types and/or participate in host immune evasion mechanisms.By sequencing seven cDNA libraries, we analyzed the MASP expression profile in trypomastigotes derived from distinct host cells and after sequential passages in acutely infected mice. Additionally, to investigate the MASP antigenic profile, we performed B-cell epitope prediction on MASP proteins and designed a MASP-specific peptide array with 110 putative epitopes, which was screened with sera from acutely infected mice.We observed differential expression of a few MASP genes between trypomastigotes derived from epithelial and myoblast cell lines. The more pronounced MASP expression changes were observed between bloodstream and tissue-culture trypomastigotes and between bloodstream forms from sequential passages in acutely infected mice. Moreover, we demonstrated that different MASP members were expressed during the acute T. cruzi infection and constitute parasite antigens that are recognized by IgG and IgM antibodies. We also found that distinct MASP peptides could trigger different antibody responses and that the antibody level against a given peptide may vary after sequential passages in mice. We speculate that changes in the large repertoire of MASP antigenic peptides during an infection may contribute to the evasion of host immune responses during the

  8. Epidemiology and Molecular Typing of Trypanosoma cruzi in Naturally-Infected Hound Dogs and Associated Triatomine Vectors in Texas, USA.

    Directory of Open Access Journals (Sweden)

    Rachel Curtis-Robles

    2017-01-01

    Full Text Available Trypanosoma cruzi is the etiologic agent of Chagas disease throughout the Americas. Few population-level studies have examined the epidemiology of canine infection and strain types of T. cruzi that infect canines in the USA. We conducted a cross-sectional study of T. cruzi infection in working hound dogs in south central Texas, including analysis of triatomine vectors collected within kennel environments.Paired IFA and Chagas Stat-Pak serological testing showed an overall seroprevalence of 57.6% (n = 85, with significant variation across kennels. Dog age had a marginally significant effect on seropositivity, with one year of age increase associated with a 19.6% increase in odds of being seropositive (odds ratio 95% CI 0.996-1.435; p = 0.055. PCR analyses of blood revealed 17.4% of dogs harbored parasite DNA in their blood, including both seronegative and seropositive dogs. Molecular screening of organs from opportunistically sampled seropositive dogs revealed parasite DNA in heart, uterus, and mammary tissues. Strain-typing showed parasite discrete typing units (DTU TcI and TcIV present in dog samples, including a co-occurrence of both DTUs in two individual dogs. Bloodmeal analysis of Triatoma gerstaeckeri and Triatoma sanguisuga insects collected from the kennels revealed exclusively dog DNA. Vector infection with T. cruzi was 80.6% (n = 36, in which T. gerstaeckeri disproportionately harbored TcI (p = 0.045 and T. sanguisuga disproportionately harbored TcIV (p = 0.029. Tracing infection status across dog litters showed some seropositive offspring of seronegative dams, suggesting infection of pups from local triatomine vectors rather than congenital transmission.Canine kennels are high-risk environments for T. cruzi transmission, in which dogs likely serve as the predominant parasite reservoir. Disease and death of working dogs from Chagas disease is associated with unmeasured yet undoubtedly significant financial consequences because working

  9. Developmental and Ultrastructural Characterization and Phylogenetic Analysis of Trypanosoma herthameyeri n. sp. of Brazilian Leptodactilydae Frogs.

    Science.gov (United States)

    Attias, Márcia; Sato, Lyslaine H; Ferreira, Robson C; Takata, Carmen S A; Campaner, Marta; Camargo, Erney P; Teixeira, Marta M G; de Souza, Wanderley

    2016-09-01

    We described the phylogenetic affiliation, development in cultures and ultrastructural features of a trypanosome of Leptodacylus chaquensis from the Pantanal biome of Brazil. In the inferred phylogeny, this trypanosome nested into the Anura clade of the basal Aquatic clade of Trypanosoma, but was separate from all known species within this clade. This finding enabled us to describe it as Trypanosoma herthameyeri n. sp., which also infects other Leptodacylus species from the Pantanal and Caatinga biomes. Trypanosoma herthameyeri multiplies as small rounded forms clumped together and evolving into multiple-fission forms and rosettes of epimastigotes released as long forms with long flagella; scarce trypomastigotes and glove-like forms are common in stationary-phase cultures. For the first time, a trypanosome from an amphibian was observed by field emission scanning electron microscopy, revealing a cytostome opening, well-developed flagellar lamella, and many grooves in pumpkin-like forms. Transmission electron microscopy showed highly developed Golgi complexes, relaxed catenation of KDNA, and a rich set of spongiome tubules in a regular parallel arrangement to the flagellar pocket as confirmed by electron tomography. Considering the basal position in the phylogenetic tree, developmental and ultrastructural data of T. herthameyeri are valuable for evolutionary studies of trypanosome architecture and cell biology. © 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.

  10. Brazilian Soybean Yields and Yield Gaps Vary with Farm Size

    Science.gov (United States)

    Jeffries, G. R.; Cohn, A.; Griffin, T. S.; Bragança, A.

    2017-12-01

    Understanding the farm size-specific characteristics of crop yields and yield gaps may help to improve yields by enabling better targeting of technical assistance and agricultural development programs. Linking remote sensing-based yield estimates with property boundaries provides a novel view of the relationship between farm size and yield structure (yield magnitude, gaps, and stability over time). A growing literature documents variations in yield gaps, but largely ignores the role of farm size as a factor shaping yield structure. Research on the inverse farm size-productivity relationship (IR) theory - that small farms are more productive than large ones all else equal - has documented that yield magnitude may vary by farm size, but has not considered other yield structure characteristics. We examined farm size - yield structure relationships for soybeans in Brazil for years 2001-2015. Using out-of-sample soybean yield predictions from a statistical model, we documented 1) gaps between the 95th percentile of attained yields and mean yields within counties and individual fields, and 2) yield stability defined as the standard deviation of time-detrended yields at given locations. We found a direct relationship between soy yields and farm size at the national level, while the strength and the sign of the relationship varied by region. Soybean yield gaps were found to be inversely related to farm size metrics, even when yields were only compared to farms of similar size. The relationship between farm size and yield stability was nonlinear, with mid-sized farms having the most stable yields. The work suggests that farm size is an important factor in understanding yield structure and that opportunities for improving soy yields in Brazil are greatest among smaller farms.

  11. Estimating Corporate Yield Curves

    OpenAIRE

    Antionio Diaz; Frank Skinner

    2001-01-01

    This paper represents the first study of retail deposit spreads of UK financial institutions using stochastic interest rate modelling and the market comparable approach. By replicating quoted fixed deposit rates using the Black Derman and Toy (1990) stochastic interest rate model, we find that the spread between fixed and variable rates of interest can be modeled (and priced) using an interest rate swap analogy. We also find that we can estimate an individual bank deposit yield curve as a spr...

  12. The miRNA and mRNA Signatures of Peripheral Blood Cells in Humans Infected with Trypanosoma brucei gambiense.

    Directory of Open Access Journals (Sweden)

    Smiths Lueong

    Full Text Available Simple, reliable tools for diagnosis of human African Trypanosomiases could ease field surveillance and enhance patient care. In particular, current methods to distinguish patients with (stage II and without (stage I brain involvement require samples of cerebrospinal fluid. We describe here an exploratory study to find out whether miRNAs from peripheral blood leukocytes might be useful in diagnosis of human trypanosomiasis, or for determining the stage of the disease. Using microarrays, we measured miRNAs in samples from Trypanosoma brucei gambiense-infected patients (9 stage I, 10 stage II, 8 seronegative parasite-negative controls and 12 seropositive, but parasite-negative subjects. 8 miRNAs (out of 1205 tested showed significantly lower expression in patients than in seronegative, parasite-negative controls, and 1 showed increased expression. There were no clear differences in miRNAs between patients in different disease stages. The miRNA profiles could not distinguish seropositive, but parasitologically negative samples from controls and results within this group did not correlate with those from the trypanolysis test. Some of the regulated miRNAs, or their predicted mRNA targets, were previously reported changed during other infectious diseases or cancer. We conclude that the changes in miRNA profiles of peripheral blood lymphocytes in human African trypanosomiasis are related to immune activation or inflammation, are probably disease-non-specific, and cannot be used to determine the disease stage. The approach has little promise for diagnostics but might yield information about disease pathology.

  13. Luminescent screens

    International Nuclear Information System (INIS)

    Lu, C.-I.

    1982-01-01

    Luminescent screens which are useful for such purposes as intensifying screens for radiographs are comprised of a support bearing a layer of finely divided particles of a phosphor dispersed in a cross-linked polymeric matrix formed by heat-curing of a coating composition comprising an unsaturated cross-linkable polymer, a polymerizable acrylic monomer, a thermoplastic polyurethane elastomer, and a heat-activatable polymerization initiator. The phosphor layer includes voids formed by evaporation of an evaporable component which is present in the coating composition from which such layer is formed. (author)

  14. Alcohol Use Screening

    Science.gov (United States)

    ... Depression Screening Substance Abuse Screening Alcohol Use Screening Alcohol Use Screening (AUDIT-C) - Instructions The following questions ... this tool, there is also text-only version . Alcohol Use Screening (AUDIT-C) - Manual Instructions The following ...

  15. Genetic and structural study of DNA-directed RNA polymerase II of Trypanosoma brucei, towards the designing of novel antiparasitic agents

    Directory of Open Access Journals (Sweden)

    Louis Papageorgiou

    2017-03-01

    Full Text Available Trypanosoma brucei brucei (TBB belongs to the unicellular parasitic protozoa organisms, specifically to the Trypanosoma genus of the Trypanosomatidae class. A variety of different vertebrate species can be infected by TBB, including humans and animals. Under particular conditions, the TBB can be hosted by wild and domestic animals; therefore, an important reservoir of infection always remains available to transmit through tsetse flies. Although the TBB parasite is one of the leading causes of death in the most underdeveloped countries, to date there is neither vaccination available nor any drug against TBB infection. The subunit RPB1 of the TBB DNA-directed RNA polymerase II (DdRpII constitutes an ideal target for the design of novel inhibitors, since it is instrumental role is vital for the parasite’s survival, proliferation, and transmission. A major goal of the described study is to provide insights for novel anti-TBB agents via a state-of-the-art drug discovery approach of the TBB DdRpII RPB1. In an attempt to understand the function and action mechanisms of this parasite enzyme related to its molecular structure, an in-depth evolutionary study has been conducted in parallel to the in silico molecular designing of the 3D enzyme model, based on state-of-the-art comparative modelling and molecular dynamics techniques. Based on the evolutionary studies results nine new invariant, first-time reported, highly conserved regions have been identified within the DdRpII family enzymes. Consequently, those patches have been examined both at the sequence and structural level and have been evaluated in regard to their pharmacological targeting appropriateness. Finally, the pharmacophore elucidation study enabled us to virtually in silico screen hundreds of compounds and evaluate their interaction capabilities with the enzyme. It was found that a series of chlorine-rich set of compounds were the optimal inhibitors for the TBB DdRpII RPB1 enzyme. All

  16. Infection rates and genotypes of Trypanosoma rangeli and T. cruzi infecting free-ranging Saguinus bicolor (Callitrichidae), a critically endangered primate of the Amazon Rainforest.

    Science.gov (United States)

    Maia da Silva, F; Naiff, R D; Marcili, A; Gordo, M; D'Affonseca Neto, J A; Naiff, M F; Franco, A M R; Campaner, M; Valente, V; Valente, S A; Camargo, E P; Teixeira, M M G; Miles, M A

    2008-08-01

    Parasites of wild primates are important for conservation biology and human health due to their high potential to infect humans. In the Amazon region, non-human primates are commonly infected by Trypanosoma cruzi and T. rangeli, which are also infective to man and several mammals. This is the first survey of trypanosomiasis in a critically endangered species of tamarin, Saguinus bicolor (Callitrichidae), from the Brazilian Amazon Rainforest. Of the 96 free-ranging specimens of S. bicolor examined 45 (46.8%) yielded blood smears positive for trypanosomes. T. rangeli was detected in blood smears of 38 monkeys (39.6%) whereas T. cruzi was never detected. Seven animals (7.3%) presented trypanosomes of the subgenus Megatrypanum. Hemocultures detected 84 positive tamarins (87.5%). Seventy-two of 84 (85.7%) were morphologically diagnosed as T. rangeli and 3 (3.1%) as T. cruzi. Nine tamarins (9.4%) yielded mixed cultures of these two species, which after successive passages generated six cultures exclusively of T. cruzi and two of T. rangeli, with only one culture remaining mixed. Of the 72 cultures positive for T. rangeli, 62 remained as established cultures and were genotyped: 8 were assigned to phylogenetic lineage A (12.9%) and 54 to lineage B (87.1%). Ten established cultures of T. cruzi were genotyped as TCI lineage (100%). Transmission of both trypanosome species, their potential risk to this endangered species and the role of wild primates as reservoirs for trypanosomes infective to humans are discussed.

  17. Characterization and Stability of Trypanosoma cruzi 24-C4 (Tc24-C4), a Candidate Antigen for a Therapeutic Vaccine Against Chagas Disease.

    Science.gov (United States)

    Biter, Amadeo B; Weltje, Sarah; Hudspeth, Elissa M; Seid, Christopher A; McAtee, C Patrick; Chen, Wen-Hsiang; Pollet, Jeroen B; Strych, Ulrich; Hotez, Peter J; Bottazzi, Maria Elena

    2018-05-01

    Chagas disease due to chronic infection with Trypanosoma cruzi is a neglected cause of heart disease, affecting approximately 6-10 million individuals in Latin America and elsewhere. T. cruzi Tc24, a calcium-binding protein in the flagellar pocket of the parasite, is a candidate antigen for an injectable therapeutic vaccine as an alternative or a complement to chemotherapy. Previously, we reported that a genetically engineered construct from which all cysteine residues had been eliminated (Tc24-C4) yields a recombinant protein with reduced aggregation and improved analytical purity in comparison to the wild-type form, without compromising antigenicity and immunogenicity. We now report that the established process for producing Escherichia coli-expressed Tc24-C4 protein is robust and reproducibly yields protein lots with consistent analytical characteristics, freeze-thaw, accelerated, and long-term stability profiles. The data indicate that, like most proteins, Tc24-C4 should be stable at -80°C, but also at 4°C and room temperature for at least 30 days, and up to 7-15 days at 37°C. Thus, the production process for recombinant Tc24-C4 is suitable for Current Good Manufacturing Practice production and clinical testing, based on process robustness, analytical characteristics, and stability profile. Copyright © 2018 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  18. Hearing Screening

    Science.gov (United States)

    Johnson-Curiskis, Nanette

    2012-01-01

    Hearing levels are threatened by modern life--headsets for music, rock concerts, traffic noises, etc. It is crucial we know our hearing levels so that we can draw attention to potential problems. This exercise requires that students receive a hearing screening for their benefit as well as for making the connection of hearing to listening.

  19. Vision Screening

    Science.gov (United States)

    ... an efficient and cost-effective method to identify children with visual impairment or eye conditions that are likely to lead ... main goal of vision screening is to identify children who have or are at ... visual impairment unless treated in early childhood. Other problems that ...

  20. Stomoxys calcitrans as possible vector of Trypanosoma evansi among camels in an affected area of the Canary Islands, Spain

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    Noé Francisco Rodríguez

    2014-07-01

    Full Text Available Introduction Trypanosoma evansi was first identified in the Canary Islands in 1997, and is still present in a small area of the Archipelago. To date, the disease has exclusively affected camel herds, and has not been detected in any other animal hosts. However potential vectors of Trypanosoma evansi must be identified. Methods One Nzi trap was placed on a camel farm located in the infected area for a period of one year. Results Two thousand five hundred and five insects were trapped, of which Stomoxys calcitrans was the sole hematophagous vector captured. Conclusions Stomoxys calcitrans could be exclusively responsible for the transmission of Trypanosoma evansi among camels in the surveyed area, as other species do not seem to be infected by S. calcitrans in the presence of camels.

  1. Status of fission yield measurements

    International Nuclear Information System (INIS)

    Maeck, W.J.

    1979-01-01

    Fission yield measurement and yield compilation activities in the major laboratories of the world are reviewed. In addition to a general review of the effort of each laboratory, a brief summary of yield measurement activities by fissioning nuclide is presented. A new fast reactor fission yield measurement program being conducted in the US is described

  2. Estudo do papel funcional da cisteína sintase e da cistationina B-sintase na resposta ao estresse oxidativo e nitrosativo em leishmania (viannia) braziliensis, trypanosoma rangeli e trypanosoma cruzi

    OpenAIRE

    Romero Calderon, Ibeth Cristina

    2014-01-01

    Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Programa de Pós-Graduação em Biotecnologia e Biociências, Florianópolis, 2014 Leishmania (Viannia) braziliensis, Trypanosoma rangeli e o Trypanosoma cruzi são parasitos hemoflagelados pertencentes à Ordem Kinetoplastida, família Trypanosomatidae, capazes de infectar insetos, animais silvestres e domésticos, assim como o homem. Durante seu ciclo de vida, estes parasitos são expostos a uma grande quanti...

  3. Enzootic transmission of Trypanosoma cruzi and T. rangeli in the Federal District of Brazil Transmissão enzoótica de Trypanosoma cruzi e T. rangeli no Distrito Federal, Brasil

    Directory of Open Access Journals (Sweden)

    Rodrigo Gurgel-Gonçalves

    2004-04-01

    Full Text Available The Federal District of Brazil (DF lies within the Cerrado biome, where open shrubland (savannas is interspersed with riverside gallery forests and permanent swamps (veredas. Trypanosoma cruzi-infected native triatomines occur in the area, but the enzootic transmission of trypanosomatids remains poorly characterized. A parasitological survey involving sylvatic triatomines (166 Rhodnius neglectus collected from Mauritia flexuosa palms and small mammals (98 marsupials and 70 rodents, totaling 18 species was conducted in 18 sites (mainly gallery forests and veredas of the DF. Parasites were isolated, morphologically identified, and characterized by PCR of nuclear (mini-exon gene and kinetoplast DNA (kDNA. Six R. neglectus, seven Didelphis albiventris and one Akodon cursor were infected by trypanosomes; wild reservoir infection is documented for the first time in the DF. kDNA PCR detected T. cruzi in five R. neglectus and mini-exon gene PCR revealed T. cruzi I in isolates from D. albiventris. Parasites infecting one bug yielded T. rangeli KP1+ kDNA amplicons. In spite of the occurrence of T. cruzi-infected D. albiventris (an important wild and peridomestic reservoir and R. neglectus (a secondary vector displaying synanthropic behavior, a low-risk of human Chagas disease transmission could be expected in the DF, considering the low prevalence infection recorded in this work. The detection of T. rangeli KP1+ associated with R. neglectus in the DF widens the known range of this parasite in Brazil and reinforces the hypothesis of adaptation of T. rangeli populations (KP1+ and KP1- to distinct evolutionary Rhodnius lineages.O Distrito Federal (DF do Brasil está localizado no bioma Cerrado, um complexo de fisionomias savânicas incluindo matas de galeria e campos úmidos permanentes (veredas. Triatomíneos silvestres infectados por Trypanosoma cruzi ocorrem na área, mas a transmissão enzoótica de tripanossomatídeos permanece insuficientemente

  4. Vision Screening

    Science.gov (United States)

    1993-01-01

    The Visi Screen OSS-C, marketed by Vision Research Corporation, incorporates image processing technology originally developed by Marshall Space Flight Center. Its advantage in eye screening is speed. Because it requires no response from a subject, it can be used to detect eye problems in very young children. An electronic flash from a 35 millimeter camera sends light into a child's eyes, which is reflected back to the camera lens. The photorefractor then analyzes the retinal reflexes generated and produces an image of the child's eyes, which enables a trained observer to identify any defects. The device is used by pediatricians, day care centers and civic organizations that concentrate on children with special needs.

  5. Utilización de Lepidium Peruvianum Maca, como medio de cultivo para el crecimiento de Trypanosoma Cruzi

    OpenAIRE

    Saldaña C, Charles; Córdova P, Ofelia; Vargas V¹, Franklin

    2006-01-01

    Por sus características nutritivas de alto valor, se ensayó la posible utilidad del Lepidium peruvianum maca, como un medio para cultivar Trypanosoma cruzi. Bajo condiciones experimentales se procedió a incubar epimastigotes de T. cruzi en cuatro medios de cultivo bifásicos diferentes, a base de Lepidium peruvianum maca, los cuales fueron comparados con el medio de cultivo BHI como control. La incorporación de maca como medio de cultivo permitió el crecimiento de Trypanosoma cruzi; se determi...

  6. Efecto inmunosupresor de la infección por Trypanosoma musculi (Mastigophora: Trypanosomatidae en la toxoplasmosis experimental Immunosuppressor effect of Trypanosoma musculi (Mastigophora: Trypanosomatidae on experimental toxoplasmosis

    Directory of Open Access Journals (Sweden)

    Loretta Piccolo-Johanning

    2013-06-01

    Full Text Available La prevalencia de infecciones por Toxoplasma gondii en el ser humano es de 5-90% según la zona geográfica; en Costa Rica por ejemplo, la seroprevalencia es de un 58%, por lo que es importante comprender algunos procesos inmunológicos, propios en estas afectaciones parasitarias. Con el objeto de determinar si el Trypanosoma musculi ejerce procesos de inmunosupresión sobre Toxoplasma gondii se realizó un experimento en el que se inocularon ratones Swiss con T. musculi cuatro, cinco, seis y siete días previos a la infección con T. gondii, ocurriendo la inmunosupresión cuando la inoculación con T. musculi fue hecha cuatro días antes. Además, la cantidad de tripomastigotos inoculados no influyó en el proceso. Se probaron tres cepas de T. gondii aisladas de las heces de un gato casero (TFC, de un Leopardus pardalis (TLP, de un Leopardus wiedii y de la carne de un Bos taurus (TBT. La cepa TLP resultó ser muy patógena, matando a los animales en un tiempo corto, independientemente de la inoculación con T. musculi; para las otras cepas se mantuvo el patrón de inmunosupresión en los ratones. Se reporta entonces un modelo experimental de inmunosupresión, aspecto muy en boga en este momento, por su relación con enfermedades que inducen esta condición en el ser humano, especialmente a enfermedades como el cáncer y el SIDA. Este modelo es más fácil de aplicar experimentalmente que el correspondiente con T. lewisi previamente descrito, el cual usa ratas blancas de más difícil manejo que los ratones usados en este estudio.The immunosuppression caused by species of the gender Trypanosoma has been widely documented. The influence over experimental infections with Toxoplasma gondii is evident when using Trypanosoma lewisi, a natural parasite of white rats. We decided to test the effect of Trypanosoma musculi from mice, an organism with very similar biological characteristics to T. lewisi, to see if this trypanosomatid could induce a similar

  7. Mode of Action of the Sesquiterpene Lactones Psilostachyin and Psilostachyin C on Trypanosoma cruzi.

    Directory of Open Access Journals (Sweden)

    Valeria P Sülsen

    Full Text Available Trypanosoma cruzi is the causative agent of Chagas' disease, which is a major endemic disease in Latin America and is recognized by the WHO as one of the 17 neglected tropical diseases in the world. Psilostachyin and psilostachyin C, two sesquiterpene lactones isolated from Ambrosia spp., have been demonstrated to have trypanocidal activity. Considering both the potential therapeutic targets present in the parasite, and the several mechanisms of action proposed for sesquiterpene lactones, the aim of this work was to characterize the mode of action of psilostachyin and psilostachyin C on Trypanosoma cruzi and to identify the possible targets for these molecules. Psilostachyin and psilostachyin C were isolated from Ambrosia tenuifolia and Ambrosia scabra, respectively. Interaction of sesquiterpene lactones with hemin, the induction of oxidative stress, the inhibition of cruzipain and trypanothione reductase and their ability to inhibit sterol biosynthesis were evaluated. The induction of cell death by apoptosis was also evaluated by analyzing phosphatidylserine exposure detected using annexin-V/propidium iodide, decreased mitochondrial membrane potential, assessed with Rhodamine 123 and nuclear DNA fragmentation evaluated by the TUNEL assay. Both STLs were capable of interacting with hemin. Psilostachyin increased about 5 times the generation of reactive oxygen species in Trypanosoma cruzi after a 4h treatment, unlike psilostachyin C which induced an increase in reactive oxygen species levels of only 1.5 times. Only psilostachyin C was able to inhibit the biosynthesis of ergosterol, causing an accumulation of squalene. Both sesquiterpene lactones induced parasite death by apoptosis. Upon evaluating the combination of both compounds, and additive trypanocidal effect was observed. Despite their structural similarity, both sesquiterpene lactones exerted their anti-T. cruzi activity through interaction with different targets. Psilostachyin

  8. Subcellular localization of glycolytic enzymes and characterization of intermediary metabolism of Trypanosoma rangeli.

    Science.gov (United States)

    Rondón-Mercado, Rocío; Acosta, Héctor; Cáceres, Ana J; Quiñones, Wilfredo; Concepción, Juan Luis

    2017-09-01

    Trypanosoma rangeli is a hemoflagellate protist that infects wild and domestic mammals as well as humans in Central and South America. Although this parasite is not pathogenic for human, it is being studied because it shares with Trypanosoma cruzi, the etiological agent of Chagas' disease, biological characteristics, geographic distribution, vectors and vertebrate hosts. Several metabolic studies have been performed with T. cruzi epimastigotes, however little is known about the metabolism of T. rangeli. In this work we present the subcellular distribution of the T. rangeli enzymes responsible for the conversion of glucose to pyruvate, as determined by epifluorescense immunomicroscopy and subcellular fractionation involving either selective membrane permeabilization with digitonin or differential and isopycnic centrifugation. We found that in T. rangeli epimastigotes the first six enzymes of the glycolytic pathway, involved in the conversion of glucose to 1,3-bisphosphoglycerate are located within glycosomes, while the last four steps occur in the cytosol. In contrast with T. cruzi, where three isoenzymes (one cytosolic and two glycosomal) of phosphoglycerate kinase are expressed simultaneously, only one enzyme with this activity is detected in T. rangeli epimastigotes, in the cytosol. Consistent with this latter result, we found enzymes involved in auxiliary pathways to glycolysis needed to maintain adenine nucleotide and redox balances within glycosomes such as phosphoenolpyruvate carboxykinase, malate dehydrogenase, fumarate reductase, pyruvate phosphate dikinase and glycerol-3-phosphate dehydrogenase. Glucokinase, galactokinase and the first enzyme of the pentose-phosphate pathway, glucose-6-phosphate dehydrogenase, were also located inside glycosomes. Furthermore, we demonstrate that T. rangeli epimastigotes growing in LIT medium only consume glucose and do not excrete ammonium; moreover, they are unable to survive in partially-depleted glucose medium. The

  9. Vector-borne transmission of Trypanosoma cruzi among captive Neotropical primates in a Brazilian zoo.

    Science.gov (United States)

    Minuzzi-Souza, Thaís Tâmara Castro; Nitz, Nadjar; Knox, Monique Britto; Reis, Filipe; Hagström, Luciana; Cuba, César A Cuba; Hecht, Mariana Machado; Gurgel-Gonçalves, Rodrigo

    2016-01-26

    Neotropical primates are important sylvatic hosts of Trypanosoma cruzi, the etiological agent of Chagas disease. Infection is often subclinical, but severe disease has been described in both free-ranging and captive primates. Panstrongylus megistus, a major T. cruzi vector, was found infesting a small-primate unit at Brasília zoo (ZooB), Brazil. ZooB lies close to a gallery-forest patch where T. cruzi circulates naturally. Here, we combine parasitological and molecular methods to investigate a focus of T. cruzi infection involving triatomine bugs and Neotropical primates at a zoo located in the Brazilian Savannah. We assessed T. cruzi infection in vectors using optical microscopy (n = 34) and nested PCR (n = 50). We used quantitative PCR (qPCR) to examine blood samples from 26 primates and necropsy samples from two primates that died during the study. We determined parasite lineages in five vectors and two primates by comparing glucose-6-phosphate isomerase (G6pi) gene sequences. Trypanosoma cruzi was found in 44 vectors and 17 primates (six genera and eight species); one Mico chrysoleucus and one Saguinus niger had high parasitaemias. Trypanosoma cruzi DNA was detected in three primates born to qPCR-negative mothers at ZooB and in the two dead specimens. One Callithrix geoffroyi became qPCR-positive over a two-year follow-up. All G6pi sequences matched T. cruzi lineage TcI. Our findings strongly suggest vector-borne T. cruzi transmission within a small-primate unit at ZooB - with vectors, and perhaps also parasites, presumably coming from nearby gallery forest. Periodic checks for vectors and parasites would help eliminate T. cruzi transmission foci in captive-animal facilities. This should be of special importance for captive-breeding programs involving endangered mammals, and would reduce the risk of accidental T. cruzi transmission to keepers and veterinarians.

  10. Synthetic Brassica napus L.: Development and Studies on Morphological Characters, Yield Attributes, and Yield

    Directory of Open Access Journals (Sweden)

    M. A. Malek

    2012-01-01

    Full Text Available Brassica napus was synthesized by hybridization between its diploid progenitor species B. rapa and B. oleracea followed by chromosome doubling. Cross with B. rapa as a female parent was only successful. Among three colchicine treatments (0.10, 0.15, and 0.20%, 0.15% gave the highest success (86% of chromosome doubling in the hybrids (AC; 2=19. Synthetic B. napus (AACC, 2=38 was identified with bigger petals, fertile pollens and seed setting. Synthetic B. napus had increased growth over parents and exhibited wider ranges with higher coefficients of variations than parents for morphological and yield contributing characters, and yield per plant. Siliqua length as well as beak length in synthetic B. napus was longer than those of the parents. Number of seeds per siliqua, 1000-seed weight and seed yield per plant in synthetic B. napus were higher than those of the parents. Although flowering time in synthetic B. napus was earlier than both parents, however the days to maturity was little higher over early maturing B. rapa parent. The synthesized B. napus has great potential to produce higher seed yield. Further screening and evaluation is needed for selection of desirable genotypes having improved yield contributing characters and higher seed yield.

  11. Caracterización molecular de los genes histona H2A y ARNsno-Cl de Trypanosoma rangeli: aplicación en pruebas diagnósticas Molecular characterization of histone H2A and snoRNA-Cl genes of Trypanosoma rangeli: application in diagnostic tests

    Directory of Open Access Journals (Sweden)

    Paula Ximena Pavía

    2009-03-01

    Full Text Available La aplicación de la reacción en cadena de la polimerasa (PCR para detectar e identificar Trypanosoma rangeli y Trypanosoma rangeli presenta a menudo dificultades de interpretación. Así, algunas pruebas generan la amplificación de bandas similares provenientes de uno de los dos parásitos, fragmentos polimórficos de un mismo parásito, o la prevalencia en la detección de T. cruzi en infecciones mixtas. En este estudio se presentan y analizan los trabajos de investigación básica realizados con el objeto de diseñar y estandarizar pruebas de PCR específicas de cada parásito. Los iniciadores TcH2AF/R se diseñaron sobre la base de la región diferencial observada entre las unidades génicas que contienen los genes h2a en estos tripanosomas. Esta pareja de iniciadores amplifican un fragmento de 234 pb específico para T. cruzi (cepas I y II. Los iniciadores TrF/R2 anillan en las regiones intergénicas del fragmento génico de 801 pb codificante para seis transcritos que forman la agrupación ARNsno-Cl en T. rangeli. Estos iniciadores amplifican un fragmento de 620 pb exclusivo de las cepas KP1(- y KP1(+ de este parásito. La aplicación de estas PCR en vectores infectados y en pacientes con enfermedad de Chagas muestra que ambas pruebas constituyen herramientas útiles para el diagnóstico y la identificación diferencial de estos tripanosomátidos.The application of polymerase chain reaction (PCR to detect Trypanosoma rangeli and Trypanosoma rangeli often presents interpretation challenges. For example, some tests yield the amplification of similar bands from either parasite, polymorphic fragments of the same parasite, or present deviation towards T. cruzi in mixed infections. In this study, the basic researching needed for designing and standardizating specific PCR tests for each parasite species PCR are shown and analyzed. The TcH2AF/R primers were designed on the basis of the differential gene region observed between the histone h2a

  12. Acerca del ciclo evolutivo del Trypanosoma (Schizotrypanum cruzi Chagas 1909, en sus fases tisular y hematica

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    Cecilio Romaña

    1956-06-01

    Full Text Available El autor pasa en revista los trabajos publicados sobre el ciclo evolutivo del Trypanosoma (S. cruzi en el huésped vertebrado, desde el descubrimiento de la enfermedad hasta nuestros días. Luego analiza las ideas de los autores modernos, fundadas en gran parte en las observaciones que ya en 1914 realizaron MAYER y ROCHA LIMA de las cuales participan actualmente ROMAÑA y MEYER, ELKELES y WOOD. Finalmente expressa que a partir de los tripanosomas infectantes los parásitos que penetram en el protoplasma celular pueden seguir dos mecanismos en su evolución hacia cuerpos leishmanioides: 1.º Por "regresión fusiforme" y 2.º por "regresión orbicular"; llegados a la forma leishmanioide los parásitos se multiplican por división binaria, una vez lleno el protoplasma celular, siguen un processo inverso de transformación hacia tripanosoma que puede seguir igualmente dos mecanismos diversos: 1. "progresión fusiforme" y 2.º "progresión orbicular". Estos diversos mecanismos de transformación están esquematizados en la fig. N.º 1 del trabajo.The author reviews published works about the evolutive cycle of the Trypanosoma cruzi in the vertebrate host, from the discovery of the disease to our days. Then, he analyzes the ideas of the modern authors who based themselves on the observations made formerly, in 1914, by MAYER & ROCHA LIMA, ideas that ROMAÑA and MEYER, ELKELES and WOOD agree at the present time. Last, he states that, from the infective trypanosomas, the parasites which enter the cellular protoplasma may follow two systems to perform their evolution up to leishmanioid bodies: 1.] by fusiform regression, 2.º by an orbicular regression. Once the parasites reach the leishmanioid forms, they multiply by binary division. When the celular protoplasm is filled up with the parasites, these follow an inverted transformation up to trypanosoma state, following also two systems; similar to the repression 1.º a fusiform progression, 2.º an

  13. The capybara (Hydrochoerus hydrochaeris) as a reservoir host for Trypanosoma evansi.

    Science.gov (United States)

    Morales, G A; Wells, E A; Angel, D

    1976-10-01

    Discovery of two ill horses and three dogs naturally infected with Trypanosoma evansi near an experimental station in the Eastern Plains of Colombia led to a search for reservoir hosts of the parasite. Infection was detected in 8/33 healthy capybaras (Hydrochoerus hydrochaeris), none of the remaining 14 horses, and none of 32 Zebu cattle (Bos indicus), 18 paca (Cuniculus paca) and 20 spiny rats (Proechimys sp.). Contrary to common opinion, the results indicated a carrier state in the capybara. Diagnosis was based on morphology, behaviour in albino rats, and pathogenicity and host range in domestic animals.

  14. Heterogeneities in the Ecoepidemiology of Trypanosoma cruzi Infection in Rural Communities of the Argentinean Chaco

    OpenAIRE

    Cardinal, M. Victoria; Orozco, M. Marcela; Enriquez, Gustavo F.; Ceballos, Leonardo A.; Gaspe, María Sol; Alvarado-Otegui, Julián A.; Gurevitz, Juan M.; Kitron, Uriel; Gürtler, Ricardo E.

    2014-01-01

    We conducted a cross-sectional survey of Trypanosoma cruzi infection of Triatoma infestans as well as dogs and cats in 327 households from a well-defined rural area in northeastern Argentina to test whether the household distribution of infection differed between local ethnic groups (Tobas and Creoles) and identify risk factors for host infection. Overall prevalence of infection of bugs (27.2%; 95% confidence interval = 25.3–29.3%), dogs (26.0%; 95% confidence interval = 23.3–30.1%), and cats...

  15. Sialoglycoconjugates in Trypanosoma cruzi-host cell interaction: possible biological model - a review

    Directory of Open Access Journals (Sweden)

    Alane Beatriz Vermelho

    1994-03-01

    Full Text Available A number of glycoconjugates, including glycolipids and glycoproteins, participate in the process of host-cell invasion by Trypanosoma cruzi and one of the most important carbohydrates involved on this interaction is sialic acid. It is known that parasite trans-sialidase participates with sialic acid in a coordinated fashion in the initial stages of invasion. Given the importance of these sialogycoconjugates, this review sets out various possible biological models for the interaction between the parasite and mammalian cells that possess a sialylated receptor/ligand system.

  16. A haptoglobin-hemoglobin receptor conveys innate immunity to Trypanosoma brucei in humans

    DEFF Research Database (Denmark)

    Vanhollebeke, Benoit; De Muylder, Géraldine; Nielsen, Marianne J

    2008-01-01

    The protozoan parasite Trypanosoma brucei is lysed by apolipoprotein L-I, a component of human high-density lipoprotein (HDL) particles that are also characterized by the presence of haptoglobin-related protein. We report that this process is mediated by a parasite glycoprotein receptor, which...... binds the haptoglobin-hemoglobin complex with high affinity for the uptake and incorporation of heme into intracellular hemoproteins. In mice, this receptor was required for optimal parasite growth and the resistance of parasites to the oxidative burst by host macrophages. In humans, the trypanosome...... immunity against the parasite....

  17. Metabolic reprogramming during the Trypanosoma brucei life cycle [version 2; referees: 4 approved

    Directory of Open Access Journals (Sweden)

    Terry K. Smith

    2017-05-01

    Full Text Available Cellular metabolic activity is a highly complex, dynamic, regulated process that is influenced by numerous factors, including extracellular environmental signals, nutrient availability and the physiological and developmental status of the cell. The causative agent of sleeping sickness, Trypanosoma brucei, is an exclusively extracellular protozoan parasite that encounters very different extracellular environments during its life cycle within the mammalian host and tsetse fly insect vector. In order to meet these challenges, there are significant alterations in the major energetic and metabolic pathways of these highly adaptable parasites. This review highlights some of these metabolic changes in this early divergent eukaryotic model organism.

  18. Metabolic reprogramming during the Trypanosoma brucei life cycle [version 1; referees: 4 approved

    Directory of Open Access Journals (Sweden)

    Terry K. Smith

    2017-05-01

    Full Text Available Cellular metabolic activity is a highly complex, dynamic, regulated process that is influenced by numerous factors, including extracellular environmental signals, nutrient availability and the physiological and developmental status of the cell. The causative agent of sleeping sickness, Trypanosoma brucei, is an exclusively extracellular protozoan parasite that encounters very different extracellular environments during its life cycle within the mammalian host and tsetse fly insect vector. In order to meet these challenges, there are significant alterations in the major energetic and metabolic pathways of these highly adaptable parasites. This review highlights some of these metabolic changes in this early divergent eukaryotic model organism.

  19. Metabolic labeling with (14C)-glucose of bloodstream and cell culture trypanosoma cruzi trypomastigotes:

    International Nuclear Information System (INIS)

    Lederkremer, R.M. de; Groisman, J.F.; Lima, C.; Katzin, A.

    1990-01-01

    Trypomastigote forms of Trypanosoma cruzi from infected mouse blood and from cell culture were metabolically labeled by incubation with D-( 14 C)-glucose. Analysis by polyacrylamide gel electrophoresis of lysates from parasites of two strains (RA and CA 1 ) showed a significantly different pattern. The difference was mainly quantitative when the blood and cell culture trypomastigotes of the RA strain were compared. Analysis of the culture medium by paper electrophoresis showed an anionic exometabolite only in the blood forms of both strains. (Author) [es

  20. Studies on the virulence and attenuation of Trypanosoma cruzi using immunodeficient animals

    Directory of Open Access Journals (Sweden)

    Basombrío Miguel Ángel

    2000-01-01

    Full Text Available Tissue invasion and pathology by Trypanosoma cruzi result from an interaction between parasite virulence and host immunity. Successive in vivo generations of the parasite select populations with increasing ability to invade the host. Conversely, prolonged in vitro selection of the parasite produces attenuated sublines with low infectivity for mammals. One such subline (TCC clone has been extensively used in our laboratory as experimental vaccine and tested in comparative experiments with its virulent ancestor (TUL. The experiments here reviewed aimed at the use of immunodeficient mice for testing the infectivity of TCC parasites. It has not been possible to obtain virulent, revertant sublines by prolonged passaged in such mice.

  1. Marker discovery in Trypanosoma vivax through GSS and comparative analysis. Preliminary data and perspectives

    International Nuclear Information System (INIS)

    Davila, A.M.R.; Guerreiro, L.T.A.; Souza, S.S.

    2005-01-01

    Trypanosoma vivax is a haemoparasite affecting the livestock industry in South America and Africa. Despite the high economic relevance of the disease caused by T. vivax, little work has been done on its molecular characterization, in contrast with human trypanosomes, such as T. brucei and T. cruzi. The present study reports the construction of a semi-normalized genomic library and the sequencing of 160 Genome Sequence Survey (GSS) ends of T. vivax. The analyses of this preliminary data show that this simple and rapid approach worked well to generate some potential new markers for this species. (author)

  2. Inositol metabolism in Trypanosoma cruzi: potential target for chemotherapy against Chagas' disease

    Directory of Open Access Journals (Sweden)

    MECIA M. OLIVEIRA

    2000-09-01

    Full Text Available Chagas' disease is a debilitating and often fatal disease caused by the protozoan parasite Trypanosoma cruzi. The great majority of surface molecules in trypanosomes are either inositol-containing phospholipids or glycoproteins that are anchored into the plasma membrane by glycosylphosphatidylinositol anchors. The polyalcohol myo-inositol is the precursor for the biosynthesis of these molecules. In this brief review, recent findings on some aspects of the molecular and cellular fate of inositol in T. cruzi life cycle are discussed and identified some points that could be targets for the development of parasite-specific therapeutic agents.

  3. Histopathologic identification of Trypanosoma cruzi (Chagas' encephalitis in an AIDS patient

    Directory of Open Access Journals (Sweden)

    Dimath Alyemni

    2017-03-01

    Full Text Available Trypanosoma cruzi (Chagas' encephalitis is an uncommon manifestation of T. cruzi infection, typically seen in immunocompromised patients. Encephalitis results from the reactivation of chronic infection predominately in individuals from endemic areas. Increased awareness of this complication is essential especially with increased migration of patients from endemic areas with concomitant HIV infection. Here we report a case of Chagas' encephalitis in an AIDS patient from Mexico in which there was no evidence of acute serologic, CSF, or blood infection by T. cruzi trypomastigotes.

  4. Presence of Trypanosoma cruzi in tissues of experimentally infected Wistar rats and their fetuses

    OpenAIRE

    Alarcón, Maritza; Lugo de Yarbuh, Ana; Moreno, Elio A; Payares, Gilberto; Araujo, Sonia; Colmenares, Melisa

    2006-01-01

    Este estudio fue realizado con un grupo de ratas juveniles hembras (Rattus norvegicus) cepa Wistar con 20 días de nacidas y 250 grs. de peso. Cada rata fue inoculada inyectándole por vía intraperitoneal 0.1 mL de la suspensión sanguínea con 1x105 tripomastigotes sanguícolas de Trypanosoma cruzi (cepa I/PAS/VE/00/PLANALTO). Los parásitos fueron aislados de Panstrongylus geniculatus, naturalmente infectado y capturado en un área urbana del valle de Caracas, Venezuela y mantenidos en ratones NMR...

  5. Avances en el estudio de la Adenilato Quinasa Nuclear de Trypanosoma cruzi

    OpenAIRE

    Cámara, María de los Milagros

    2012-01-01

    Trypanosoma cruzi, el agente etiológico del Mal de Chagas es un eucariota inferior en donde el control de la expresión génica recae mayormente en mecanismos postraduccionales. Durante todo su ciclo de vida se observan fluctuaciones en la expresión génica. En la presente tesis se realizó el estudio de una adenilato quinasa nuclear (TcADKn) que se encuentra involucrada en la biogénesis ribosomal. Las adenilato quinasas nucleares han sido descriptas en muy pocos organismos, se las ha asociado al...

  6. Effect of Integrated Nutrient Management on Yield and Yield ...

    African Journals Online (AJOL)

    Declining soil fertility is one of the major problems causing yield reduction of barley ... (VC) with inorganic NP on growth, yield and yield components of food barley. ... The experiments were laid out in a randomized complete block design with ...

  7. Plants used in Guatemala for the treatment of protozoal infections: II. Activity of extracts and fractions of five Guatemalan plants against Trypanosoma cruzi.

    Science.gov (United States)

    Berger, I; Barrientos, A C; Cáceres, A; Hernández, M; Rastrelli, L; Passreiter, C M; Kubelka, W

    1998-09-01

    The activities of crude plant extracts of five plants popularly used in Guatemala against bacterial and protozoal infections and some of their fractions have been evaluated against the trypomastigote and epimastigote forms of Trypanosoma cruzi in vitro. The most active fraction of Neurolaena lobata has also been screened in vivo. Main in vitro activities against trypomastigotes have been observed for the hexane and ethanol extracts of N. lobata (Asteraceae). Both extracts were also active against epimastigotes, whereas all other extracts tested had no effect on epimastigotes. For the hexane extracts of Petiveria alliacea (Phytolaccaceae) and Tridax procumbens (Asteraceae) a marked inhibition of trypomastigotes has been found. Also the ethanol extracts of Byrsonima crassifolia (Malpighiaceae) leafs and Gliricidia sepium (Papilionaceae) bark showed some trypanocidal activity. Fraction 2 of the ethanol extract of N. lobata was highly active against T. cruzi as well in vitro as in vivo. The chloroforme fraction of P. alliacea showed a high inhibition of trypomastigotes in vitro. Also three fractions of the active extract of B. crassifolia inhibited T. cruzi trypomastigotes. No fraction of G. sepium bark extract showed a marked trypanocidal activity.

  8. Yield enhancement with DFM

    Science.gov (United States)

    Paek, Seung Weon; Kang, Jae Hyun; Ha, Naya; Kim, Byung-Moo; Jang, Dae-Hyun; Jeon, Junsu; Kim, DaeWook; Chung, Kun Young; Yu, Sung-eun; Park, Joo Hyun; Bae, SangMin; Song, DongSup; Noh, WooYoung; Kim, YoungDuck; Song, HyunSeok; Choi, HungBok; Kim, Kee Sup; Choi, Kyu-Myung; Choi, Woonhyuk; Jeon, JoongWon; Lee, JinWoo; Kim, Ki-Su; Park, SeongHo; Chung, No-Young; Lee, KangDuck; Hong, YoungKi; Kim, BongSeok

    2012-03-01

    A set of design for manufacturing (DFM) techniques have been developed and applied to 45nm, 32nm and 28nm logic process technologies. A noble technology combined a number of potential confliction of DFM techniques into a comprehensive solution. These techniques work in three phases for design optimization and one phase for silicon diagnostics. In the DFM prevention phase, foundation IP such as standard cells, IO, and memory and P&R tech file are optimized. In the DFM solution phase, which happens during ECO step, auto fixing of process weak patterns and advanced RC extraction are performed. In the DFM polishing phase, post-layout tuning is done to improve manufacturability. DFM analysis enables prioritization of random and systematic failures. The DFM technique presented in this paper has been silicon-proven with three successful tape-outs in Samsung 32nm processes; about 5% improvement in yield was achieved without any notable side effects. Visual inspection of silicon also confirmed the positive effect of the DFM techniques.

  9. Water screen

    Energy Technology Data Exchange (ETDEWEB)

    Kutepov, A.I.; Fedotov, I.N.; Prokopov, O.I.

    1981-01-01

    The invention refers to ventilation and can be used for repair-fitting operations in a blasting-dangerous gas condition, for example, during elimination of gas-oil gushers, repair of gas-oil pipelines, equipment etc. In order to improve safety of labor, the nozzle adapters of the water collector are oriented towards each other. The collector is installed on a support with the possibility of rotating and vertical movement. The proposed screen excludes the possibility of blasting-dangerous concentrations of gases and guarantees extinguishing of the impact spark during operation of the tool.

  10. RBC Antibody Screen

    Science.gov (United States)

    ... C Cystic Fibrosis (CF) Gene Mutations Testing Cytomegalovirus (CMV) Tests D-dimer Dengue Fever Testing Des-gamma- ... Index of Screening Recommendations Not Listed? Not Listed? Newborn Screening Screening Tests for Infants Screening Tests for ...

  11. Mental Health Screening Center

    Science.gov (United States)

    ... Releases & Announcements Public Service Announcements Partnering with DBSA Mental Health Screening Center These online screening tools are not ... you have any concerns, see your doctor or mental health professional. Depression Screening for Adult Depression Screening for ...

  12. Breast cancer screening

    Science.gov (United States)

    Mammogram - breast cancer screening; Breast exam - breast cancer screening; MRI - breast cancer screening ... is performed to screen women to detect early breast cancer when it is more likely to be cured. ...

  13. No gold standard estimation of the sensitivity and specificity of two molecular diagnostic protocols for Trypanosoma brucei spp. in Western Kenya.

    Directory of Open Access Journals (Sweden)

    Barend Mark de Clare Bronsvoort

    2010-01-01

    Full Text Available African animal trypanosomiasis is caused by a range of tsetse transmitted protozoan parasites includingTrypanosoma vivax, Trypanosoma congolense and Trypansoma brucei. In Western Kenya and other parts of East Africa two subspecies of T. brucei, T.b. brucei and the zoonoticT.b. rhodesiense, co-circulate in livestock. A range of polymerase chain reactions (PCR have been developed as important molecular diagnostic tools for epidemiological investigations of T. brucei s.l. in the animal reservoir and of its zoonotic potential. Quantification of the relative performance of different diagnostic PCRs is essential to ensure comparability of studies. This paper describes an evaluation of two diagnostic test systems for T. brucei using a T. brucei s.l. specific PCR [1] and a single nested PCR targeting the Internal Transcribed Spacer (ITS regions of trypanosome ribosomal DNA [2]. A Bayesian formulation of the Hui-Walter latent class model was employed to estimate their test performance in the absence of a gold standard test for detecting T.brucei s.l. infections in ear-vein blood samples from cattle, pig, sheep and goat populations in Western Kenya, stored on Whatman FTA cards. The results indicate that the system employing the T. brucei s.l. specific PCR (Se1=0.760 had a higher sensitivity than the ITS-PCR (Se2=0.640; both have high specificity (Sp1=0.998; Sp2=0.997. The true prevalences for livestock populations were estimated (pcattle=0.091, ppigs=0.066, pgoats=0.005, psheep=0.006, taking into account the uncertainties in the specificity and sensitivity of the two test systems. Implications of test performance include the required survey sample size; due to its higher sensitivity and specificity, the T. brucei s.l. specific PCR requires a consistently smaller sample size than the ITS-PCR for the detection of T. brucei s.l. However the ITS-PCR is able to simultaneously screen samples for other pathogenic trypanosomes and may thus be, overall, a better

  14. A note on hypoplastic yielding

    OpenAIRE

    Nader, José Jorge

    2010-01-01

    This note discusses briefly the definition of yield surface in hypoplasticity in connection with the physical notion of yielding. The relation of yielding with the vanishing of the material time derivative of the stress tensor and the vanishing of the corotational stress rate is investigated.

  15. Insight into the exoproteome of the tissue-derived trypomastigote form of trypanosoma cruzi

    DEFF Research Database (Denmark)

    Queiroz, Rayner M L; Ricart, Carlos A O; Machado, Mara O

    2016-01-01

    The protozoan parasite Trypanosoma cruzi causes Chagas disease, one of the major neglected infectious diseases. It has the potential to infect any nucleated mammalian cell. The secreted/excreted protein repertoire released by T. cruzi trypomastigotes is crucial in host-pathogen interactions...

  16. Structural model of a putrescine-cadaverine permease from Trypanosoma cruzi predicts residues vital for transport and ligand binding

    NARCIS (Netherlands)

    Soysa, R.; Venselaar, H.; Poston, J.; Ullman, B.; Hasne, M.P.

    2013-01-01

    The TcPOT1.1 gene from Trypanosoma cruzi encodes a high affinity putrescine-cadaverine transporter belonging to the APC (amino acid/polyamine/organocation) transporter superfamily. No experimental three-dimensional structure exists for any eukaryotic member of the APC family, and thus the structural

  17. THE CYTOSOLIC AND GLYCOSOMAL GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE FROM TRYPANOSOMA-BRUCEI - KINETIC-PROPERTIES AND COMPARISON WITH HOMOLOGOUS ENZYMES

    NARCIS (Netherlands)

    LAMBEIR, AM; LOISEAU, AM; KUNTZ, DA; VELLIEUX, FM; MICHELS, PAM; OPPERDOES, FR

    1991-01-01

    The protozoan haemoflagellate Trypanosoma brucei has two NAD-dependent glyceraldehyde-3-phosphate dehydrogenase isoenzymes, each with a different localization within the cell. One isoenzyme is found in the cytosol, as in other eukaryotes, while the other is found in the glycosome, a microbody-like

  18. Mitochondrial translation factors of Trypanosoma brucei: elongation factor-Tu has a unique subdomain that is essential for its function

    Czech Academy of Sciences Publication Activity Database

    Cristodero, M.; Mani, J.; Oeljeklaus, S.; Aeberhard, L.; Hashimi, Hassan; Ramrath, D.J.F.; Lukeš, Julius; Warscheid, B.; Schneider, A.

    2013-01-01

    Roč. 90, č. 4 (2013), s. 744-755 ISSN 0950-382X R&D Projects: GA ČR GAP305/12/2261 Institutional support: RVO:60077344 Keywords : mitochondrial translation * Trypanosoma brucei * EF-Tu Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.026, year: 2013

  19. Adaptations in the glucose metabolism of procyclic Trypanosoma brucei isolates from Tsetse flies and during differentiation of bloodstream forms.

    NARCIS (Netherlands)

    van Grinsven, K.W.A.; van den Abbeele, J.; van den Bossche, P.; van Hellemond, J.J.; Tielens, A.G.M.

    2009-01-01

    Procyclic forms of Trypanosoma brucei isolated from the midguts of infected tsetse flies, or freshly transformed from a strain that is close to field isolates, do not use a complete Krebs cycle. Furthermore, short stumpy bloodstream forms produce acetate and are apparently metabolically preadapted

  20. Phylogeny and morphological variability of trypanosomes from African pelomedusid turtles with redescription of Trypanosoma mocambicum Pienaar, 1962

    Czech Academy of Sciences Publication Activity Database

    Dvořáková, N.; Čepička, I.; Qablan, M. A.; Gibson, W.; Blažek, Radim; Široký, P.

    2015-01-01

    Roč. 166, č. 6 (2015), s. 599-608 ISSN 1434-4610 Institutional support: RVO:68081766 Keywords : Trypanosoma * turtle * Pelusios * polymorphism * phylogeny * SSU rRNA gene Subject RIV: EG - Zoology Impact factor: 2.898, year: 2015

  1. Effect of stage of maturity on dry matter yield, morphological ...

    African Journals Online (AJOL)

    The experiment evaluated effect of stage of maturity on dry matter yield, morphological characteristics and nutritive value of burgundy bean (Macroptilium bracteatum) at the screen house of Department of Agronomy, Bayero University Kano, Nigeria. The treatments were 3 stages of growth repeated 3 times in a completely ...

  2. Evaluation of polycross sweetpotato seedlings for root yield potential ...

    African Journals Online (AJOL)

    The study aimed at determining the root yield potential of the sweetpotato seedlings, the variation in storage root flesh colour and response of the storage roots to major pests and diseases attacking sweetpotato in the field .The experiment was carried out in the screen house and at the Eastern experimental field of National ...

  3. Trypanosoma cruzi: strain selection by diferent schedules of mouse passage of an initially mixed infection

    Directory of Open Access Journals (Sweden)

    Maria P. Deane

    1984-12-01

    Full Text Available From an initial double infection in mice, established by simultaneous and equivalent inocula of bloodstream forms of strains Y and F of Trypanosoma cruzi, two lines were derived by subinoculations: one (W passaged every week, the other (M every month. Through biological and biochemical methods only the Y strain was identified at the end of the 10th and 16th passages of line W and only the F strain at the 2nd and 4th passages of line M. The results illustrate strain selection through laboratory manipulation of initially mixed populations of T. cruzi.De uma infecção inicialmente dupla em camundongo, estabelecida por inóculo simultaneo e equivalente de formas sanguíneas das cepas Y e F de Trypanosoma cruzi, duas linhagens foram originadas por subinoculações: uma (W passada casa semana, a outra (M cada mês. Por métodos biológicos e bioquímicos apenas a cepa Y foi identificada ao fim a 10a. e 16a. passagens da linhagem W e apenas a cepa F na 2a. e 4a.passagens de linhagem M. Os resultados demonstram a seleção de cepas através de manipulação em laboratorio de populações inicialmente mistas de T. cruzi.

  4. Moderate physical exercise protects myenteric metabolically more active neurons in mice infected with Trypanosoma cruzi.

    Science.gov (United States)

    Moreira, Neide Martins; de Moraes, Solange Marta Franzói; Dalálio, M M O; Gomes, Mônica Lúcia; Sant'ana, D M G; de Araújo, Silvana Marques

    2014-02-01

    Trypanosoma cruzi causes neuronal myenteric depopulation compromising intestinal function. The purpose of this study was to evaluate the influence of moderate physical exercise on NADH diaphorase (NADH-d)-positive neurons in the myenteric plexus and intestinal wall of the colon in mice infected with T. cruzi. Forty 30-day-old male Swiss mice were divided into the following groups: trained infected (TI), sedentary infected (SI), trained control (TC), and sedentary control. The TC and TI groups were subjected to a moderate physical exercise program on a treadmill for 8 weeks. Three days after finishing physical exercise, the TI and SI groups were intraperitoneally inoculated with 1,300 blood trypomastigotes of the Y strain of Trypanosoma cruzi. Parasitemia was evaluated from days 4 to 61 after inoculation. On day 75 of infection, myenteric neurons in the colon were quantified (NADH-d), and inflammatory foci were counted. Tumor necrosis factor-α (TNF-α) and transforming growth factor-β (TGF-β) levels were evaluated in plasma. The results were compared using analysis of variance and the Kruskal-Wallis test at a 5 % significance level. Moderate physical exercise reduced the parasite peak on day 8 of infection (p = 0.0132) and total parasitemia (p = 0.0307). It also prevented neuronal depopulation (p  0.05). These results reinforce the therapeutic benefits of moderate physical exercise for T. cruzi infection.

  5. In vivo trypanocidal activity of Nymphaea lotus Linn. methanol extract against Trypanosoma brucei brucei

    Directory of Open Access Journals (Sweden)

    Muhammad Haruna Garba

    2015-10-01

    Full Text Available Objective: To evaluate the antitrypanosomal potentials of methanol extract of Nymphaea lotus Linn. (N. lotus with the aim of obtaining a new lead for formulating safe, inexpensive, nontoxic and readily available trypanocidal drugs. Methods: Seventy percent (v/v (methanol/water crude extract of N. lotus was evaluated for antitrypanosomal activity in experimental trypanosomiasis using Trypanosoma brucei bruceiinfected mice. Infected mice in different groups were administered intraperitoneally 100, 200, 300 and 400 mg/kg body weight/day of the crude for two weeks, while a positive control group was treated with standard drug, berenil. Results: The crude extract at a dose of 100 mg/kg body weight/day was more effective than the higher doses in completely clearing parasites from the blood of mice infected with Trypanosoma brucei brucei. Pre-treatment of healthy mice with the crude extract for 5 days before infection did not prevent the establishment of the infection, indicating that the extract had no prophylactic activity. Subinoculation of the blood and cerebrospinal fluid drawn from the cured mice into healthy mice failed to produce any infection within 50 days post inoculation. Administration of 1 000 mg/kg body weight of the crude extract led to the death of 50% of the experimental animals indicating a high level of toxicity of the extract at higher doses. Conclusions: This study has demonstrated the potency of the crude extract of N. lotus in treating experimental trypanosomiasis at lower doses.

  6. Investigating the Chaperone Properties of a Novel Heat Shock Protein, Hsp70.c, from Trypanosoma brucei

    Directory of Open Access Journals (Sweden)

    Adélle Burger

    2014-01-01

    Full Text Available The neglected tropical disease, African Trypanosomiasis, is fatal and has a crippling impact on economic development. Heat shock protein 70 (Hsp70 is an important molecular chaperone that is expressed in response to stress and Hsp40 acts as its co-chaperone. These proteins play a wide range of roles in the cell and they are required to assist the parasite as it moves from a cold blooded insect vector to a warm blooded mammalian host. A novel cytosolic Hsp70, from Trypanosoma brucei, TbHsp70.c, contains an acidic substrate binding domain and lacks the C-terminal EEVD motif. The ability of a cytosolic Hsp40 from Trypanosoma brucei J protein 2, Tbj2, to function as a co-chaperone of TbHsp70.c was investigated. The main objective was to functionally characterize TbHsp70.c to further expand our knowledge of parasite biology. TbHsp70.c and Tbj2 were heterologously expressed and purified and both proteins displayed the ability to suppress aggregation of thermolabile MDH and chemically denatured rhodanese. ATPase assays revealed a 2.8-fold stimulation of the ATPase activity of TbHsp70.c by Tbj2. TbHsp70.c and Tbj2 both demonstrated chaperone activity and Tbj2 functions as a co-chaperone of TbHsp70.c. In vivo heat stress experiments indicated upregulation of the expression levels of TbHsp70.c.

  7. Trans-sialidase inhibition assay detects Trypanosoma cruzi infection in different wild mammal species.

    Science.gov (United States)

    Sartor, Paula A; Ceballos, Leonardo A; Orozco, Marcela M; Cardinal, Marta V; Gürtler, Ricardo E; Leguizamón, María S

    2013-08-01

    The detection of Trypanosoma cruzi infection in mammals is crucial for understanding the eco-epidemiological role of the different species involved in parasite transmission cycles. Xenodiagnosis (XD) and hemoculture (HC) are routinely used to detect T. cruzi in wild mammals. Serological methods are much more limited because they require the use of specific antibodies to immunoglobulins of each mammalian species susceptible to T. cruzi. In this study we detected T. cruzi infection by trans-sialidase (TS) inhibition assay (TIA). TIA is based on the antibody neutralization of a recombinant TS that avoids the use of anti-immunoglobulins. TS activity is not detected in the co-endemic protozoan parasites Leishmania spp and T. rangeli. In the current study, serum samples from 158 individuals of nine wild mammalian species, previously tested by XD, were evaluated by TIA. They were collected from two endemic areas in northern Argentina. The overall TIA versus XD co-reactivity was 98.7% (156/158). All 18 samples from XD-positive mammals were TIA-positive (co-positivity, 100%) and co-negativity was 98.5% (138/140). Two XD-negative samples from a marsupial (Didelphis albiventris) and an edentate (Dasypus novemcinctus) were detected by TIA. TIA could be used as a novel tool for serological detection of Trypanosoma cruzi in a wide variety of sylvatic reservoir hosts.

  8. The flagellum of Trypanosoma brucei: new tricks from an old dog

    Science.gov (United States)

    Ralston, Katherine S.; Hill, Kent L.

    2010-01-01

    African trypanosomes, i.e. Trypanosoma brucei and related sub-species, are devastating human and animal pathogens that cause significant human mortality and limit sustained economic development in sub-Saharan Africa. Trypanosoma brucei is a highly motile protozoan parasite and coordinated motility is central to both disease pathogenesis in the mammalian host and parasite development in the tsetse fly vector. Since motility is critical for parasite development and pathogenesis, understanding unique aspects of the T. brucei flagellum may uncover novel targets for therapeutic intervention in African sleeping sickness. Moreover, studies of conserved features of the T. brucei flagellum are directly relevant to understanding fundamental aspects of flagellum and cilium function in other eukaryotes, making T. brucei an important model system. The T. brucei flagellum contains a canonical 9 + 2 axoneme, together with additional features that are unique to kinetoplastids and a few closely-related organisms. Until recently, much of our knowledge of the structure and function of the trypanosome flagellum was based on analogy and inference from other organisms. There has been an explosion in functional studies in T. brucei in recent years, revealing conserved as well as novel and unexpected structural and functional features of the flagellum. Most notably, the flagellum has been found to be an essential organelle, with critical roles in parasite motility, morphogenesis, cell division and immune evasion. This review highlights recent discoveries on the T. brucei flagellum. PMID:18472102

  9. Molecular profiles of Venezuelan isolates of Trypanosoma sp. by random amplified polymorphic DNA method.

    Science.gov (United States)

    Perrone, T M; Gonzatti, M I; Villamizar, G; Escalante, A; Aso, P M

    2009-05-12

    Nine Trypanosoma sp. Venezuelan isolates, initially presumed to be T. evansi, were collected from three different hosts, capybara (Apure state), horse (Apure state) and donkey (Guarico state) and compared by the random amplification polymorphic DNA technique (RAPD). Thirty-one to 46 reproducible fragments were obtained with 12 of the 40 primers that were used. Most of the primers detected molecular profiles with few polymorphisms between the seven horse, capybara and donkey isolates. Quantitative analyses of the RAPD profiles of these isolates revealed a high degree of genetic conservation with similarity coefficients between 85.7% and 98.5%. Ten of the primers generated polymorphic RAPD profiles with two of the three Trypanosoma sp. horse isolates, namely TeAp-N/D1 and TeGu-N/D1. The similarity coefficient between these two isolates and the rest, ranged from 57.9% to 68.4% and the corresponding dendrogram clustered TeAp-N/D1 and Te Gu-N/D1 in a genetically distinct group.

  10. Trypanosoma cf. varani in an imported ball python (Python reginus) from Ghana.

    Science.gov (United States)

    Sato, Hiroshi; Takano, Ai; Kawabata, Hiroki; Une, Yumi; Watanabe, Haruo; Mukhtar, Maowia M

    2009-08-01

    Peripheral blood from a ball python (Python reginus) imported from Ghana was cultured in Barbour-Stoenner-Kelly (BSK) medium for Borrelia spp. isolation, resulting in the prominent appearance of free, and clusters of, trypanosomes in a variety of morphological forms. The molecular phylogenetic characterization of these cultured trypanosomes, using the small subunit rDNA, indicated that this python was infected with a species closely related to Trypanosoma varani Wenyon, 1908, originally described in the Nile monitor lizard (Varanus niloticus) from Sudan. Furthermore, nucleotide sequences of glycosomal glyceraldehyde-3-phosphate dehydrogenase gene of both isolates showed few differences. Giemsa-stained blood smears, prepared from the infected python 8 mo after the initial observation of trypanosomes in hemoculture, contained trypomastigotes with a broad body and a short, free flagellum; these most closely resembled the original description of T. varani, or T. voltariae Macfie, 1919 recorded in a black-necked spitting cobra (Naja nigricollis) from Ghana. It is highly possible that lizards and snakes could naturally share an identical trypanosome species. Alternatively, lizards and snakes in the same region might have closely related, but distinct, Trypanosoma species as a result of sympatric speciation. From multiple viewpoints, including molecular phylogenetic analyses, reappraisal of trypanosome species from a wide range of reptiles in Africa is needed to clarify the relationship of recorded species, or to unmask unrecorded species.

  11. Molecular epidemiology of Trypanosoma cruzi and Triatoma dimidiata in costal Ecuador.

    Science.gov (United States)

    Wong, Yim Yan; Sornosa Macias, Karen Jeniffer; Guale Martínez, Doris; Solorzano, Luis F; Ramirez-Sierra, Maria Jesus; Herrera, Claudia; Dumonteil, Eric

    2016-07-01

    Chagas disease is a neglected tropical disease caused by the protozoan parasite Trypanosoma cruzi. In Ecuador, Triatoma dimidiata and Rhodnius ecuadoriensis are the main vector species, responsible for over half of the cases of T. cruzi infection in the country. T. dimidiata is believed to have been introduced in Ecuador during colonial times, and its elimination from the country is thus believed to be feasible. We investigated here the molecular ecology of T. dimidiata and T. cruzi in costal Ecuador to further guide control efforts. Analysis of the Internal Transcribed Spacer 2 (ITS-2) of 23 specimens from Progreso, Guayas, unambiguously supported the likely importation of T. dimidiata from Central America to Ecuador. The observation of a very high parasite infection rate (54%) and frequent feeding on humans (3/5) confirmed a continued risk of transmission to humans. All genotyped parasites corresponded to TcI DTU and Trypanosoma rangeli was not detected in T. dimidiata. TcI subgroups corresponded to TcIa (25%), and mixed infections with TcIa and TcId (75%). Further studies should help clarify T. cruzi genetic structure in the country, and the possible impact of the introduction of T. dimidiata on the circulating parasite strains. The elevated risk posed by this species warrants continuing efforts for its control, but its apparent mobility between peridomestic and domestic habitats may favor reinfestation following insecticide spraying. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Plants of Brazilian restingas with tripanocide activity against Trypanosoma cruzi strains.

    Science.gov (United States)

    Faria, Robson Xavier; Souza, André Luis Almeida; Lima, Barbara; Tietbohl, Luis Armando Candido; Fernandes, Caio Pinho; Amaral, Raquel Rodrigues; Ruppelt, Bettina Monika; Santos, Marcelo Guerra; Rocha, Leandro

    2017-12-01

    Chagas disease is caused by the Trypanosoma cruzi affecting millions of people, and widespread throughout Latin America. This disease exhibits a problematic chemotherapy. Benznidazole, which is the drug currently used as standard treatment, lamentably evokes several adverse reactions. Among other options, natural products have been tested to discover a novel therapeutic drug for this disease. A lot of plants from the Brazilian flora did not contain studies about their biological effects. Restinga de Jurubatiba from Brazil is a sandbank ecosystem poorly studied in relation to plant biological activity. Thus, three plant species from Restinga de Jurubatiba were tested against in vitro antiprotozoal activity. Among six extracts obtained from leaves and stem parts and 2 essential oils derived from leave parts, only 3 extracts inhibited epimastigote proliferation. Substances present in the extracts with activity were isolated (quercetin, myricetin, and ursolic acid), and evaluated in relation to antiprotozoal activity against epimastigote Y and Dm28 Trypanosoma cruzi strains. All isolated substances were effective to reduce protozoal proliferation. Essentially, quercetin and myricetin did not cause mammalian cell toxicity. In summary, myricetin and quercetin molecule can be used as a scaffold to develop new effective drugs against Chagas's disease.

  13. The MORPHEUS II protein crystallization screen

    International Nuclear Information System (INIS)

    Gorrec, Fabrice

    2015-01-01

    MORPHEUS II is a 96-condition initial crystallization screen formulated de novo. The screen incorporates reagents selected from the Protein Data Bank to yield crystals that are not observed in traditional conditions. In addition, the formulation facilitates the optimization and cryoprotection of crystals. High-quality macromolecular crystals are a prerequisite for the process of protein structure determination by X-ray diffraction. Unfortunately, the relative yield of diffraction-quality crystals from crystallization experiments is often very low. In this context, innovative crystallization screen formulations are continuously being developed. In the past, MORPHEUS, a screen in which each condition integrates a mix of additives selected from the Protein Data Bank, a cryoprotectant and a buffer system, was developed. Here, MORPHEUS II, a follow-up to the original 96-condition initial screen, is described. Reagents were selected to yield crystals when none might be observed in traditional initial screens. Besides, the screen includes heavy atoms for experimental phasing and small polyols to ensure the cryoprotection of crystals. The suitability of the resulting novel conditions is shown by the crystallization of a broad variety of protein samples and their efficiency is compared with commercially available conditions

  14. The MORPHEUS II protein crystallization screen

    Energy Technology Data Exchange (ETDEWEB)

    Gorrec, Fabrice, E-mail: fgorrec@mrc-lmb.cam.ac.uk [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH (United Kingdom)

    2015-06-27

    MORPHEUS II is a 96-condition initial crystallization screen formulated de novo. The screen incorporates reagents selected from the Protein Data Bank to yield crystals that are not observed in traditional conditions. In addition, the formulation facilitates the optimization and cryoprotection of crystals. High-quality macromolecular crystals are a prerequisite for the process of protein structure determination by X-ray diffraction. Unfortunately, the relative yield of diffraction-quality crystals from crystallization experiments is often very low. In this context, innovative crystallization screen formulations are continuously being developed. In the past, MORPHEUS, a screen in which each condition integrates a mix of additives selected from the Protein Data Bank, a cryoprotectant and a buffer system, was developed. Here, MORPHEUS II, a follow-up to the original 96-condition initial screen, is described. Reagents were selected to yield crystals when none might be observed in traditional initial screens. Besides, the screen includes heavy atoms for experimental phasing and small polyols to ensure the cryoprotection of crystals. The suitability of the resulting novel conditions is shown by the crystallization of a broad variety of protein samples and their efficiency is compared with commercially available conditions.

  15. Epidemiology and Molecular Typing of Trypanosoma cruzi in Naturally-Infected Hound Dogs and Associated Triatomine Vectors in Texas, USA

    Science.gov (United States)

    Curtis-Robles, Rachel; Snowden, Karen F.; Dominguez, Brandon; Dinges, Lewis; Rodgers, Sandy; Mays, Glennon

    2017-01-01

    Background Trypanosoma cruzi is the etiologic agent of Chagas disease throughout the Americas. Few population-level studies have examined the epidemiology of canine infection and strain types of T. cruzi that infect canines in the USA. We conducted a cross-sectional study of T. cruzi infection in working hound dogs in south central Texas, including analysis of triatomine vectors collected within kennel environments. Methodology/Principle Findings Paired IFA and Chagas Stat-Pak serological testing showed an overall seroprevalence of 57.6% (n = 85), with significant variation across kennels. Dog age had a marginally significant effect on seropositivity, with one year of age increase associated with a 19.6% increase in odds of being seropositive (odds ratio 95% CI 0.996–1.435; p = 0.055). PCR analyses of blood revealed 17.4% of dogs harbored parasite DNA in their blood, including both seronegative and seropositive dogs. Molecular screening of organs from opportunistically sampled seropositive dogs revealed parasite DNA in heart, uterus, and mammary tissues. Strain-typing showed parasite discrete typing units (DTU) TcI and TcIV present in dog samples, including a co-occurrence of both DTUs in two individual dogs. Bloodmeal analysis of Triatoma gerstaeckeri and Triatoma sanguisuga insects collected from the kennels revealed exclusively dog DNA. Vector infection with T. cruzi was 80.6% (n = 36), in which T. gerstaeckeri disproportionately harbored TcI (p = 0.045) and T. sanguisuga disproportionately harbored TcIV (p = 0.029). Tracing infection status across dog litters showed some seropositive offspring of seronegative dams, suggesting infection of pups from local triatomine vectors rather than congenital transmission. Conclusions/Significance Canine kennels are high-risk environments for T. cruzi transmission, in which dogs likely serve as the predominant parasite reservoir. Disease and death of working dogs from Chagas disease is associated with unmeasured yet

  16. A rapid method for testing in vivo the susceptibility of different strains of Trypanosoma cruzi to active chemotherapeutic agents

    Directory of Open Access Journals (Sweden)

    Leny S. Filardi

    1984-06-01

    Full Text Available A method is described which permits to determine in vivo an in a short period of time (4-6 hours the sensitivity of T. cruzo strains to known active chemotherapeutic agents. By using resistant- and sensitive T. cruzi stains a fairly good correlation was observed between the results obtained with this rapid method (which detects activity against the circulating blood forms and those obtained with long-term schedules which involve drug adminstration for at least 20 consecutive days and a prolonged period of assessment. This method may be used to characterize susceptibility to active drugs used clinically, provide infomation on the specific action against circulating trypomastigotes and screen active compounds. Differences in the natural susceptibility of Trypanosoma cruzi strains to active drugs have been already reported using different criteria, mostly demanding long-term study of the animal (Hauschka, 1949; Bock, Gonnert & Haberkorn, 1969; Brener, Costa & Chiari, 1976; Andrade & Figueira, 1977; Schlemper, 1982. In this paper we report a method which detects in 4-6 hours the effect of drugs on bloodstream forms in mice with established T. cruzi infections. The results obtained with this method show a fairly good correlation with those obtained by prolonged treatment schedules used to assess the action of drugs in experimental Chagas' disease and may be used to study the sensitivity of T. cruzi strains to active drugs.No presente trabalho descreve-se um metodo que permite determinar in vivo e em curto espaço de tempo (4-6 horas a sensibilidade de cepas de T. cruzi a agentes terapeuticos ativos na doença de Chagas. Usando-se cepas sensíveis e resistentes aos medicamentos foi possível observar uma boa correlação entre os resultados obtidos com o método rápido (que detecta atividade contra as formas circulantes do parasita e aqueles obtidos com esquema de acao prolongada que envolve a administração da droga por 20 dias e posterior avalia

  17. Prevalence of antibodies to Trypanosoma cruzi, Toxoplasma gondii, Encephalitozonn cuniculi, Sarcocystis neurona, Besnoitia darlingi, and Neospora caninum in North American opossum, Didelphis virginiana, from Southern Louisian

    Science.gov (United States)

    We examined the prevalence of antibodies to zoonotic protozoan parasites (Trypanosoma cruzi, Toxoplasma gondii, and Encephalitozoon cuniculi) and protozoan’s of veterinary importance (Neospora caninum, Sarcocystis neurona and Besnoitia darlingi) in a population of North American opossums (Didelphis...

  18. Trypanosoma teixeirae: A new species belonging to the T. cruzi clade causing trypanosomosis in an Australian little red flying fox (Pteropus scapulatus).

    Science.gov (United States)

    Barbosa, Amanda D; Mackie, John T; Stenner, Robyn; Gillett, Amber; Irwin, Peter; Ryan, Una

    2016-06-15

    Little is known about the genetic diversity and pathogenicity of trypanosomes in Australian bats. Recently a novel trypanosome species was identified in an adult female little red flying fox (Pteropus scapulatus) with clinical and pathological evidence of trypanosomosis. The present study used morphology and molecular methods to demonstrate that this trypanosome is a distinct species and we propose the name Trypanosoma teixeirae sp. n. Morphological comparison showed that its circulating trypomastigotes were significantly different from those of Trypanosoma pteropi and Trypanosoma hipposideri, two species previously described from Australian bats. Genetic information was not available for T. pteropi and T. hipposideri but phylogenetic analyses at the 18S ribosomal RNA (rRNA) and glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) loci indicated that T. teixeirae sp. n. was genetically distinct and clustered with other bat-derived trypanosome species within the Trypanosoma cruzi clade. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. [Early stages of development of Trypanosoma rotatorium (Mayer, 1843) from peripheral blood and internal organs of Anurans Bufo bufo (Linnaeus) and Rana sp. (Anura)].

    Science.gov (United States)

    Malysheva, M N

    2014-01-01

    The data on the fauna of trypanosomes of Anura of the Leningrad Province are given. The initial development stages of Trypanosoma rotatorium in peripheral blood and internal organs of the frog are described for the first time.

  20. Characterization of a Novel Class I Transcription Factor A (CITFA) Subunit That Is Indispensable for Transcription by the Multifunctional RNA Polymerase I of Trypanosoma brucei

    KAUST Repository

    Nguyen, T. N.; Nguyen, B. N.; Lee, J. H.; Panigrahi, A. K.; Gunzl, A.

    2012-01-01

    Trypanosoma brucei is the only organism known to have evolved a multifunctional RNA polymerase I (pol I) system that is used to express the parasite's ribosomal RNAs, as well as its major cell surface antigens, namely, the variant surface

  1. On the tissular parasitism of Trypanosoma cruzi y strain in swiss mice Sobre o parasitismo tecidual da cepa Y do Trypanosoma cruzi em camundongos albinos (Swiss-Webster

    Directory of Open Access Journals (Sweden)

    Maria Auxiliadora de Sousa

    1984-12-01

    Full Text Available A review of the tissular parasitism of Trypanosoma cruzi Y strain in Swiss mice was carried out. This strain parasitized preferentially smooth, skeletal and cardiac muscle fibers, with low transitory spleen and liver parasitism, as previously found by some Authors, although differing from other reports. These results can be related to the host genetical constitution and/or the degree of the strain virulence at the time of this study. Furthermore, we discuss that the high macrophagotropism reported for this strain in some instances could be an artificially induced condition resulting from its serial maintenance in mice, either for a longer time and/or by using young animals. The heavy parasitism and inflammation observed in the bladder, pancreas and spermatic duct of some inoculated mice, as well as the testis parasitization, were also noteworthy findings.Através deste trabalho fizemos uma revisão do parasitismo tecidual da cepa Y do Trypanosoma cruzi em camundongos albinos (Swiss-Webster. Esta cepa parasitou preferencialmente as fibras musculares lisas, esqueléticas e cardíacas, sendo baixo e transitório seu parasitismo do baço e fígado, conforme já observado por alguns Autores, embora diferindo de outros achados. Estes resultados podem estar relacionados com o padrão genético do hospedeiro e/ou com o grau de virulência da cepa por ocasião deste estudo. Além do mais, discutimos a possibilidade de que o intenso macrofagotropismo descrito para esta cepa em algumas ocasiões possa ser uma condição artificialmente induzida através de sua manutenção seriada em camundongos por tempo prolongado e/ou pelo uso de animais jovens. Também são dignos de nota, o intenso parasitismo e inflamação da bexiga, pâncreas e canal espermático de alguns animais inoculados, assim como, o encontro de ninhos de amastigotas no testículo.

  2. Effect of experimental single Ancylostoma caninum and mixed infections of Trypanosoma brucei and Trypanosoma congolense on the humoural immune response to anti-rabies vaccination in dogs

    Directory of Open Access Journals (Sweden)

    Nwoha Rosemary Ijeoma Ogechi

    2015-06-01

    Full Text Available Objective: To determine the effect of Ancylostoma caninum (A. caninum and trypanosome parasites on the immune response to vaccination in dogs in endemic environments. Methods: Sixteen dogs for the experiment were grouped into 4 of 4 members each. Group I was the uninfected control one, and GPII was infected with A. caninum; GPIII was infected with A. caninum/Trypanosoma congolense (T. congolense, and GPIV was infected with Trypanosoma brucei (T. brucei/A. caninum. The dogs were first vaccinated with antirabies vaccine before infecting GPII, GPIII and GPIV with A. caninum which were done 4 weeks after vaccination. By 2-week post-vaccination, trypanosome parasites were superimposed on both GPIII and GPIV. A secondary vaccination was given to GPI, GPII, GPIII, and GPIV by Week 12 of the experiment (4 weeks post treatment. Results: The prepatent period was (3.00 ± 1.40 days, in the conjunct infection of T. brucei/ A. caninum. It was (9.00 ± 1.10 days, in conjunct T. congolense/A. caninum. The prepatent period of A. caninum was (14.0 ± 2.0 days in the single A. caninum group and (13.0 ± 1.0 days in the conjunct trypanosome/A. caninum. At the 1st week after vaccination, the antibody titer in all the vaccinated groups (GPI, GPII, GPIII, and GPIV significantly increased (P < 0.05 and peaked at the 3rd week after vaccination. Following infections, there were marked significant decreases (P < 0.05 in the antibody production against rabies in GPII, GPIII and GPIV. The significant decrease (P < 0.05 in antibody titer was highest in the conjunct groups (GPIII and GPIV compared to the single infection (GPII. Treatment with diminazene aceturate and mebendazole did not significantly improve antibody response in the dogs. A secondary vaccination administered at the 12th week after the primary vaccination significantly increased (P < 0.05 the antibody titer with a peak at the 3rd week after the secondary vaccination. Conclusions: It was therefore concluded

  3. Humoral immune response of horses experimentally infected with Trypanosoma evansi/ Resposta imune humoral de eqüinos infectados experimentalmente com Trypanosoma evansi

    Directory of Open Access Journals (Sweden)

    Lúcia Padilha Cury Thomaz de Aquino

    2001-05-01

    Full Text Available Six adult horses were experimentally infected with Trypanosoma evansi (106 parasites. Three other adult horses served as negative control. Serum samples of the experimentally infected horses with T. evansi and non-infected controls horses were obtained before inoculation, and daily thereafter until 14 days post infection (DPI. After that time the serum samples were obtained weekly. Sera of the infected and non-infected control horses was tested by indirect fluorescent antibody test (IFAT and enzyme-linked immunosorbent assay (ELISA for the detection of antibodies against T. evansi. Both ELISA and IFAT detected trypanosomal antibodies shortly after infection and showed progressive increases in antibodies levels during early stages of infection. The responses started on the eighth and eleventh DPI. Maximum IFAT and ELISA values were reached after four weeks of infection and were maintained at this level until the end of the period of study.Seis eqüinos foram inoculados com 106 tripomastigota sangüícolas de Trypanosoma evansi. Três outros animais foram mantidos como testemunhas. Amostras de soro sangüíneo foram obtidas de todos os animais, antes da inoculação, e diariamente até o 14º dia pós inoculação (DPI; após este período uma vez por semana. Pesquisa de anticorpos anti- T. evansi, foram realizadas através da reação de imunofluorescência indireta (RIFI e do ensaio de imunoabsorção enzimática (ELISA. A resposta imune humoral, detectada através da RIFI e do ELISA, iniciou-se, em média, a partir do oitavo DPI, alcançando títulos máximos após quatro semanas de evolução, e os titulos de anticorpos anti- T. evansi mantiveram-se elevadas até o término das observações.

  4. Estudios sobre Trypanosoma rangeli Tejera, 1920: VIII. Respuesta a las reinfecciones en dos mamíferos Trypanosoma rangeli Tejera, 1920: VIII. Responses to reinfections in 2 mammals

    Directory of Open Access Journals (Sweden)

    N. Añez

    1985-06-01

    Full Text Available Bajo condiciones experimentales se estudia el curso de la infección primaria y la respuesta a las reinfecciones por Trypanosoma rangeli en ratones albinos y Didelphis marsupialis. Durante el curso de la infección primaria en ratones, se observa una parasitemia relativamente baja y de corta duración. Los mismos muestran durante la primera reinfección una parasitemia escasa de cuatro días de duración, siendo resistentes a las sucesivas reinfecciones con T. rangeli. Los ejemplares de D. marsupialis exhiben una parasitemia de más larga duración, pero con un nivel de parásitos sanguícolas mucho menor que el detectado en el modelo ratón, siendo la respuesta a las reinfecciones similar a la observada en ratones. Se detectan anticuerpos hemaglutinantes en los sueros inmunes de ratones y Didelphis marsupialis, sometidos a la reinfección por T. rangeli. Se especula sobre la posible acción sinérgica de una respuesta inmune en el sitio de deposición en contra de las formas metacíclicas de T. rangeli y la acción de anticuerpos circulantes en contra de las formas sanguícolas, para explicar la resistencia de ambos modelos a las reinfecciones por T. rangeli.Under experimental conditions, the course of the infection and the response to the reinfection by Trypanosoma rangeli in mice and Didelphis marsupialis, are studied. During the initial infection the mice show a relatively low parasitaemia and a short patent period. A scanty parasitaemia level of four days length, was observed following the first reinfection, being the mice resistant to new reinfections by T. rangeli. In opossums a lower parasitaemia and a longer patent period than that detected in mice, were observed during the initial infection. The response to reinfections in this mammal, was similar to that observed in mice. After reinfection with T. rangeli, haemagglutinant antibodies in immune-sera of both mice and opossums, were detected. The possible immune-response at the site of

  5. Systematics in delayed neutron yields

    Energy Technology Data Exchange (ETDEWEB)

    Ohsawa, Takaaki [Kinki Univ., Higashi-Osaka, Osaka (Japan). Atomic Energy Research Inst.

    1998-03-01

    An attempt was made to reproduce the systematic trend observed in the delayed neutron yields for actinides on the basis of the five-Gaussian representation of the fission yield together with available data sets for delayed neutron emission probability. It was found that systematic decrease in DNY for heavier actinides is mainly due to decrease of fission yields of precursors in the lighter side of the light fragment region. (author)

  6. Retinopathy of prematurity screening criteria in Iran: new screening guidelines.

    Science.gov (United States)

    Roohipoor, Ramak; Karkhaneh, Reza; Farahani, Afsar; Ebrahimiadib, Nazanin; Modjtahedi, Bobeck; Fotouhi, Akbar; Yaseri, Mehdi; Khodabande, Alireza; Zarei, Mohammad; Imani Fuladi, Marjan; Taheri, Arash; Riazi Esfahani, Mohammad; Loewenstein, John

    2016-07-01

    To test the applicability of existing retinopathy of prematurity (ROP) guidelines on Iranian patients and to develop novel ROP screening criteria in Iran. Both eyes of 1932 infants born ≤37 weeks of gestation and/or weighting ≤3000 g were included in this prospective cohort study that was conducted across nine neonatal intensive care units and a tertiary eye hospital ROP clinic. The patients were examined for ROP and the need for treatment (type 1 ROP or worse). All the patients were screened 4 weeks after birth or at 31 weeks of postmenstrual age, whichever was later. The patients were followed until retinal vascularisation was completed or the patients reached 50 weeks of gestational age (GA) without prethreshold ROP. A receiver operating characteristic curve was used to determine the best screening criteria for ROP. Screening criteria from other countries were applied to our patient data to determine their ability to appropriately detect ROP. Patients with ROP requiring treatment. The mean GA±SD and birth weight (BW)±SD of the screened patients were 32±2.7 weeks and 1713±516 g, respectively. Using criteria of GA≤32 weeks or BW ≤2000 yielded sensitivity and specificity of 100% and 26.7%, respectively, for treatment requiring ROP regardless of clinical comorbidities. Using screening recommendations of American Academy of Pediatrics would miss 25.4% of ROP and 8.4%ROP requiring treatment in our cohort. Other countries screening recommendations would result in a significant amount of missed cases of treatment requiring ROP when applied to Iran. As a result, we have proposed new guidelines for premature babies in Iran. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  7. VARIABILITY OF YIELD AND YIELD COMPONENTS IN “EGUSI ...

    African Journals Online (AJOL)

    journal

    Estimate of expected genetic advance in seed yield plant-1 ranged between. 25.90-48.40%. ..... values in fruit and seed yield characters have been reported in culinary melon, ... and Khund, A. 2004. Extent of heterosis and heritability in some.

  8. Response of Yield and Yield Components of Tef [Eragrostis Tef ...

    African Journals Online (AJOL)

    The partial budget analysis also indicates that applications of 46 kg. N ha-1 and 10 kg P ha-1 are ..... (1994) indicated that where the grain yield response is negative, yield reduction is primarily caused by a .... An Economic Training. Manual.

  9. On yield gaps and yield gains in intercropping

    NARCIS (Netherlands)

    Gou, Fang; Yin, Wen; Hong, Yu; Werf, van der Wopke; Chai, Qiang; Heerink, Nico; Ittersum, van Martin K.

    2017-01-01

    Wheat-maize relay intercropping has been widely used by farmers in northwest China, and based on field experiments agronomists report it has a higher productivity than sole crops. However, the yields from farmers’ fields have not been investigated yet. Yield gap analysis provides a framework to

  10. 7755 EFFECT OF NPK FERTILIZER ON FRUIT YIELD AND YIELD ...

    African Journals Online (AJOL)

    Win7Ent

    2013-06-03

    Jun 3, 2013 ... peasant farmers in Nigeria. With the increased ... did not significantly (p=0.05) increase the fruit yield nor the seed yield. Key words: NPK fertilizer, Fruit ..... SAS (Statistical Analysis System) Version 9.1. SAS Institute Inc., Cary, ...

  11. SLIFER measurement for explosive yield

    International Nuclear Information System (INIS)

    Bass, R.C.; Benjamin, B.C.; Miller, H.M.; Breding, D.R.

    1976-04-01

    This report describes the shorted location indicator by frequency of electrical resonance (SLIFER) system used at Sandia Laboratories for determination of explosive yield of under ground nuclear tests

  12. Infections of Hypostomus spp. by Trypanosoma spp. and leeches: a study of hematology and record of these hirudineans as potential vectors of these hemoflagellates

    Directory of Open Access Journals (Sweden)

    Lincoln Lima Corrêa

    Full Text Available Abstract Among Kinetoplastida, the Trypanosoma is the genus with the highest occurrence infecting populations of marine fish and freshwater in the world, with high levels of prevalence, causing influences fish health and consequent economic losses, mainly for fish populations in situation stress. This study investigated infections of Hypostomus spp. by Trypanosoma spp. and leeches, as well as blood parameters of this host in the network of tributaries of the Tapajós River in the state of Pará, in the eastern Amazon region in Brazil. Of the 47 hosts examined, 89.4% were parasitized by Trypanosoma spp. and 55.4% also had leeches attached around the mouth. The intensity of Trypanosoma spp. increased with the size of the host, but the body conditions were not influenced by the parasitism. The number of red blood cells, and hemoglobin, mean corpuscular volume (MCV, mean corpuscular hemoglobin concentration (MCHC, mean corpuscular hemoglobin (MCH, total number of leukocytes and thrombocytes showed variations and negative correlation with the intensity of Trypanosoma spp. in the blood of the hosts. The results suggest that the leeches were vectors of Trypanosoma spp. in Hypostomus spp.

  13. Effects of betamethasone on the course of experimentai. Infection with Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Frederico G.C. Abath

    1986-09-01

    Full Text Available In this experiment, the effect of betamethasone administered in the early post- acute infection of mice by Trypanosoma cruzi was studied. This drug was administered during 30 days after the 42nd day of infection in a dose of 0.15 mg/day. The betamethasone treatment did not cause fresh outbreaks of parasitemia and the histopathological findings in the chronic phase were not different from those in the control group. The higher cumulative mortality after treatment in the experimental group was due to superimposed bacterial infections. Outbred albino mice infected with low numbers ofY strain Trypanosoma cruzi trypomastigotes were not suitable models for Chagas' disease, since after 7 months of observation only mild histological lesions developed in all the animais. Prolonged betamethasone treatment of mice infected with low numbers o/Trypanosoma cruzi of the Y strain, during the post-acute phase did not aggravate the course of infection.Foram estudados os efeitos da betametasona administrada na fase pós-aguda imediata de uma infecção pelo T. cruzi em camundongos. O tratamento consistiu de 30 doses diárias de 0,15 mg de betametasona, a partir de 42° dia de infecção, não havendo aparecimento de novos surtos de parasitemia. No tempo de duração do experimento (7 meses não houve diferença entre as lesões histopatológicas dos animais tratados e dos não tratados. O grupo experimental apresentou uma maior mortalidade acumulada no 75º dia de infecção, o que pode ser atribuído a infecções bacterianas associadas. Por outro lado, camundongos albinos "outbred", infectados com baixo inóculo, não se apresentaram como bom modelo de doença de Chagas, já que não desenvolveram lesões importantes nem na fase aguda nem após 7 meses de infecção. Em conclusão, o tratamento imunosupressivo prolongado, após a fase aguda de uma infecção mínima com a cepa Ydo T. cruzi não tem influência sobre o curso da infecção, pelo menos no que tange

  14. Factors that determine the effectiveness of screening for congenital heart malformations at child health centres

    NARCIS (Netherlands)

    R.E. Juttmann (Rikard); J. Hess (Jakob); C.W.N. Looman (Caspar); P.J. van der Maas (Paul)

    2000-01-01

    textabstractBACKGROUND: The actual yield from current screening for clinically significant congenital heart malformations in Dutch child health care is far from optimal. In this study factors that determine the effectiveness of this screening are identified and

  15. Overdiagnosis in organised mammography screening in Denmark. A comparative study

    DEFF Research Database (Denmark)

    Jørgensen, Karsten J; Zahl, Per-Henrik; Gøtzsche, Peter C

    2009-01-01

    % of the Danish population has been offered organised mammography screening over a long time-period. METHODS: We collected incidence rates of carcinoma in situ and invasive breast cancer in areas with and without screening over 13 years with screening (1991-2003), and 20 years before its introduction (1971...... an overdiagnosis of 35% when we compared unadjusted incidence rates for the screened and non-screened areas, but after compensating for a small decline in incidence in older, previously screened women. Our adjusted Poisson regression analysis indicated a relative risk of 1.40 (95% CI: 1.35-1.45) for the whole...... screening period, and a potential compensatory drop in older women of 0.90 (95% CI: 0.88-0.96), yielding an overdiagnosis of 33%, which we consider the most reliable estimate. The drop in previously screened women was only present in one of the two screened regions and was small in absolute numbers...

  16. Unraveling the differences of the hydrolytic activity of Trypanosoma cruzi trans-sialidase and Trypanosoma rangeli sialidase: a quantum mechanics-molecular mechanics modeling study.

    Science.gov (United States)

    Bueren-Calabuig, Juan A; Pierdominici-Sottile, Gustavo; Roitberg, Adrian E

    2014-06-05

    Chagas' disease, also known as American trypanosomiasis, is a lethal, chronic disease that currently affects more than 10 million people in Central and South America. The trans-sialidase from Trypanosoma cruzi (T. cruzi, TcTS) is a crucial enzyme for the survival of this parasite: sialic acids from the host are transferred to the cell surface glycoproteins of the trypanosome, thereby evading the host's immune system. On the other hand, the sialidase of T. rangeli (TrSA), which shares 70% sequence identity with TcTS, is a strict hydrolase and shows no trans-sialidase activity. Therefore, TcTS and TrSA represent an excellent framework to understand how different catalytic activities can be achieved with extremely similar structures. By means of combined quantum mechanics-molecular mechanics (QM/MM, SCC-DFTB/Amberff99SB) calculations and umbrella sampling simulations, we investigated the hydrolysis mechanisms of TcTS and TrSA and computed the free energy profiles of these reactions. The results, together with our previous computational investigations, are able to explain the catalytic mechanism of sialidases and describe how subtle differences in the active site make TrSA a strict hydrolase and TcTS a more efficient trans-sialidase.

  17. Breeding for Grass Seed Yield

    DEFF Research Database (Denmark)

    Boelt, Birte; Studer, Bruno

    2010-01-01

    Seed yield is a trait of major interest for many fodder and amenity grass species and has received increasing attention since seed multiplication is economically relevant for novel grass cultivars to compete in the commercial market. Although seed yield is a complex trait and affected...... by agricultural practices as well as environmental factors, traits related to seed production reveal considerable genetic variation, prerequisite for improvement by direct or indirect selection. This chapter first reports on the biological and physiological basics of the grass reproduction system, then highlights...... important aspects and components affecting the seed yield potential and the agronomic and environmental aspects affecting the utilization and realization of the seed yield potential. Finally, it discusses the potential of plant breeding to sustainably improve total seed yield in fodder and amenity grasses....

  18. Fertility of the Small East African goat following pre-pubertal infection with Trypanosoma congolense

    International Nuclear Information System (INIS)

    O'Hara, H.B.; Gombe, S.

    1991-01-01

    Pre-pubertal male and female Small East African goats were infected with Trypanosoma congolense at 4-5 months of age. Changes in body weight and haemogram were monitored weekly. Progesterone and testosterone measurements were made three times weekly until the goats either reached puberty or 18 months of age. Onset of puberty was determined from observation of oestrus behaviour, mating or increase in libidio; this was confirmed by elevation in plasma progesterone or testosterone levels. Trypanosomiasis affected pre-pubertal goats by reducing body weight gain and delaying onset of puberty. Histological examination of the gonads showed pronounced pathological changes. These effects were reversed by treatment with isometamidium chloride (Samorin, May and Baker). It was concluded that early treatment of infected goats before serious gonadal damage could occur allowed full restoration of reproductive function. (author). 6 refs, 4 figs, 1 tab

  19. A new bianthron glycoside as inhibitor of Trypanosoma cruzi glyceraldehyde 3-phosphate dehydrogenase activity

    International Nuclear Information System (INIS)

    Macedo, Edangelo M.S. de; Silva, Maria G.V.; Wiggers, Helton J.; Montanari, Carlos A.; Braz-Filho, Raimundo; Andricopulo, Adriano D.

    2009-01-01

    A phytochemical investigation of the ethanolic extract of stalks of Senna martiana Benth. (Leguminoseae), native specie of northeast Brazil, resulted in the isolation and spectroscopic characterization of a new bianthrone glycoside, martianine 1 (10,10'-il-chrysophanol-10-oxi- 10,10'-bi-glucosyl). Its identification was established by HRMS, IR and 2D NMR experiments. The evaluation of martianine trypanocidal activity was carried out against gliceraldehyde 3-phosphate dehydrogenase enzyme from Trypanosoma cruzi. Its inhibitory constant (K i ) is in the low micromolar concentration and it was determined by isothermal titration calorimetry to be 27.3 +-2.47 μmol L -1 . The non-competitive mechanism is asserted to be putative of the mode of action martianine displays against T. cruzi GAPDH. Results show that martianine has a great potential to become new lead molecule by inhibiting this key enzyme and for the development of new drugs against Chagas disease. (author)

  20. Activity of D-carnitine and its derivatives on Trypanosoma infections in rats and mice

    Directory of Open Access Journals (Sweden)

    Manganaro M.

    2003-06-01

    Full Text Available Little progress has been made in the treatment of African trypanosomiasis over the past decades. L-carnitine has a major role in glycolysis-based energy supply of blood trypanosomes for it stimulates constant ATP production. To investigate whether administration of the isomer D-carnitine could exert a competitive inhibition on the metabolic pathway of the L-form, possibily resulting in parasite replication inhibition, several formulations of this compound were tested on Trypanosoma lewisi and T. brucei rhodesiense in rodent models. High oral dosages of D-carnitine inner salt and proprionyl-D-carnitine were not toxic to animals and induced about 50 % parasite growth inhibition in reversible, i.e. competitive, fashion. A putative mechanism could be an interference in pyruvate kinase activity and hence ATP production. Considering both, lack of toxicity and inhibitory activity, D-carnitine may have a role in the treatment of African trypanosomiasis, in association with available trypanocidal drugs.

  1. Genetic characterization of Trypanosoma cruzi natural clones from the state of Paraíba, Brazil

    Directory of Open Access Journals (Sweden)

    Christian Barnabé

    2005-05-01

    Full Text Available Eighteen Trypanosoma cruzi stocks from the state of Paraíba, Brazil, isolated from man, wild mammals, and triatomine bugs were studied by multilocus enzyme electrophoresis and random primed amplified polymorphic DNA. Despite the low number of stocks, a notable genetic, genotypic, and phylogenetic diversity was recorded. The presence of the two main phylogenetic subdivisions, T. cruzi I and II, was recorded. The strong linkage disequilibrium observed in the population under survey suggests that T. cruzi undergoes predominant clonal evolution in this area too, although this result should be confirmed by a broader sample. The pattern of clonal variation does not suggests a recent origin by founder effect with a limited number of different genotypes.

  2. Prevalencia de infeccion a Trypanosoma cruzi en donadores de sangre en el Estado de Jalisco, Mexico

    Directory of Open Access Journals (Sweden)

    Francisco Trujillo Contreras

    1993-06-01

    Full Text Available Durante el periodo de Octubre de 1991 a Marzo de 1992, se tomaron 3419 muestras de donadores de sangre de 12 localidades rurales y de 8 hospitales urbanos a los que se les realizo un estúdio serológico mediante la reacción de hemaglutinación indirecta encontrándose anticuerpos contra Trypanosoma cruzi en 44 indivíduos 39 masculinosy 5 femininos. El 90,9% de donantes fueron masculinos. De acuerdo a su procedencia, el 73,5% fué del área urbana y el 26,5% del área rural. De acuerdo a los resultados el riesgo de transmisión de T. cruzi por transfusión sanguinea está latente por la creciente urbanización de la enfermedad de Chagas.

  3. Production and expression of inflammation and angiogenic parameters triggered by different genetic population of Trypanosoma cruzi.

    OpenAIRE

    Shrestha, Deena

    2014-01-01

    Programa de Pós-Graduação em Ciências Biológicas. Núcleo de Pesquisas em Ciências Biológicas, Pró-Reitoria de Pesquisa e Pós Graduação, Universidade Federal de Ouro Preto. A cardiopatia induzida pela infecção pelo Trypanosoma cruzi aprensenta a inflamação como sua principal característica imunopatológica. Differente células inflamatórias contribuem para a produção de mediatores inflamatorios e regulatórios promotores diretos ou indiretos do processo denominado angiogênese inflamatória. As ...

  4. Genómica del Trypanosoma cruzi. Nuevas oportunidades para tratar el mal de Chagas

    Directory of Open Access Journals (Sweden)

    Jorge A. Huete-Pérez

    2006-12-01

    Full Text Available LA SECUENCIACIÓN DEL GENOMA HUMANO PUBLICADA EN FEBRERO de 2001 ha sido considerada como el hito científico más importante del siglo XX. La secuenciación, cuatro años más tarde, de tres parásitos tripanosmatidas, entre ellos el Trypanosoma cruzi, podría ser también catalogada como uno de los acontecimientos científicos más importantes para la salud publica del continente americano. Aquí se presenta un panorama general sobre los resultados más significativos del estudio geonómico del T. cruzi, se abordan los trabajos realizados por nuestro laboratorio en la Universidad Centroamericana, finalizando con una discusión sobre las perspectivas del uso de la genómica en Nicaragua.

  5. Effects of water deprivation on renal hydroelectrolytic excretion in chronically Trypanosoma cruzi-infected rats

    Directory of Open Access Journals (Sweden)

    T.T. Rosa

    1995-03-01

    Full Text Available The effect of an 8 hour-period of water deprivation on fluid and electrolyte renal excretion was investigated in male Wistar rats infected with the strain São Felipe (12SF of Trypanosoma cruzi, in comparison with age and sex matched non-infected controls. The median percent reductions in the urinary flow (-40% v -63% and excretion ofsodium (-57% v-79% were smaller in chagasic than in control rats, respectively. So, chagasic rats excreted more than controls. On the other hand, the median percent decrement in the clearance of creatinine was higher in chagasic (-51% than in controls (-39%. Thus, chagasic rats showed some disturbed renal hydroelectrolytic responses to water deprivation, expressed by smaller conservation, or higher excretion of water and sodium in association with smaller glomerularfiltration rate. This fact denoted an elevation in the fractional excretion of sodium and water.

  6. [Seroprevalence of Trypanosoma cruzi infection in the rural population of Sucre State, Venezuela].

    Science.gov (United States)

    García-Jordán, Noris; Berrizbeitia, Mariolga; Rodríguez, Jessicca; Concepción, Juan Luis; Cáceres, Ana; Quiñones, Wilfredo

    2017-10-26

    The current study aimed to determine the seroprevalence of Trypanosoma cruzi infection in Sucre State, Venezuela, and its association with epidemiological risk factors. The cluster sampling design allowed selecting 96 villages and 576 dwellings in the State's 15 municipalities. A total of 2,212 serum samples were analyzed by ELISA, HAI, and IFI. Seroprevalence in Sucre State was 3.12%. Risk factors associated with T. cruzi infection were: accumulated garbage, flooring and wall materials, type of dwelling, living in a house with wattle and daub walls and/or straw roofing, living in a house with risky walls and roofing, risky buildings and wattle and daub outbuildings, poultry inside the human dwelling, and presence of firewood. Infection was associated with individual age, and three seropositive cases were found in individuals less than 15 years of age. Sucre State has epidemiological factors that favor the risk of acquiring T. cruzi infection.

  7. CHARACTERIZATION AND ANTIPARASITIC ACTIVITY OF BENZOPHENONE THIOSEMICARBAZONES ON Trypanosoma brucei brucei

    Directory of Open Access Journals (Sweden)

    Georges C. Accrombessi

    2011-02-01

    Full Text Available The structure of four synthesized thiosemicarbazones, substituted or not, of benzophenone has been confirmed by spectrometrical analysis IR, NMR 1H and 13C. Their anti-trypanosomal activities were evaluated on Trypanosoma brucei brucei. Among these compounds, benzophenone 4 phenyl-3-thiosemicarbazone 4 has the highest activity with the half-inhibitory concentration (IC50 = 8.48 micromolar (µM. Benzophenone 4-methyl-3-thiosemicarbazone 3 and benzophenone thiosemicarbazone 1 showed moderate anti-trypanosomal activity with IC50 values equal to 23.27 µM and 67.17 µM respectively. Benzophenone 2 methyl-3-thiosemicarbazone 2 showed no activity up to IC50 = 371.74 µM.

  8. Enhanced succinic acid production in Aspergillus saccharolyticus by heterologous expression of fumarate reductase from Trypanosoma brucei

    DEFF Research Database (Denmark)

    Yang, Lei; Lübeck, Mette; Ahring, Birgitte K.

    2015-01-01

    production medium as well as the complete medium, but the measured enzyme activities were different depending on the media. Furthermore, a soluble NADH-dependent fumarate reductase gene (frd) from Trypanosoma brucei was inserted and expressed in A. saccharolyticus. The expression of the frd gene led......Aspergillus saccharolyticus exhibits great potential as a cell factory for industrial production of dicarboxylic acids. In the analysis of the organic acid profile, A. saccharolyticus was cultivated in an acid production medium using two different pH conditions. The specific activities...... of the enzymes, pyruvate carboxylase (PYC), malate dehydrogenase (MDH), and fumarase (FUM), involved in the reductive tricarboxylic acid (rTCA) branch, were examined and compared in cells harvested from the acid production medium and a complete medium. The results showed that ambient pH had a significant impact...

  9. Seroprevalence of human Trypanosoma cruzi infection in diferent geografic zones of Chiapas, Mexico.

    Science.gov (United States)

    Mazariego-Arana, M A; Monteón, V M; Ballinas-Verdugo, M A; Hernández-Becerril, N; Alejandre-Aguilar, R; Reyes, P A

    2001-01-01

    A serologic survey was carried out in four different geographic zones of Chiapas, Mexico. A total of 1,333 samples were collected from residents of thirteen communities located on the Coast, Central Mountain, Lacandon Forest and a zone called Mesochiapas. One hundred and fifty one seropositive individuals (11.3%) were identified. Human Trypanosoma cruzi infection was influenced by geography. In the Lacandon Forest and Central Mountains there was a higher seroprevalence 32.1 and 13.8% respectively, than on the coast (1.2%). In Mesochiapas there were no seropositive individuals among the 137 persons tested. An active transmission is probably continuing because seropositive cases (13.8%) were detected in children under 10 years of age. The vector recognized on the Coast was Triatoma dimidiata while in the Lacandon Forest it was Rhodnius prolixus.

  10. Population genetic analysis of Colombian Trypanosoma cruzi isolates revealed by enzyme electrophoretic profiles

    Directory of Open Access Journals (Sweden)

    Manuel Ruiz-Garcia

    2001-01-01

    Full Text Available Although Colombia presents an enormous biological diversity, few studies have been conducted on the population genetics of Trypanosoma cruzi. This study was carried out with 23 Colombian stocks of this protozoa analyzed for 13 isoenzymatic loci. The Hardy-Weinberg equilibrium, the genetic diversity and heterogeneity, the genetic relationships and the possible spatial structure of these 23 Colombian stocks of T. cruzi were estimated. The majority of results obtained are in agreement with a clonal population structure. Nevertheless, two aspects expected in a clonal structure were not discovered in the Colombian T. cruzi stocks. There was an absence of given zymodemes over-represented from a geographical point of view and the presumed temporal stabilizing selective phenomena was not observed either in the Colombian stocks sampled several times through the years of the study. Some hypotheses are discussed in order to explain the results found.

  11. African trypanosomiasis with special reference to Egyptian Trypanosoma evansi: is it a neglected zoonosis?

    Science.gov (United States)

    El-Bahnasawy, Mamdouh M M; Khater, Mai Kh A; Morsy, Tosson A

    2014-12-01

    Trypanosomes (including humans) are blood and sometimes tissue parasites of the order Kinetoplastida, family Trypanosomatidae, genus Trypanosoma, principally transmitted by biting insects where most of them undergo a biological cycle. They are divided into Stercoraria with the posterior station inoculation, including T. cruzi, both an extra- and intracellular parasite that causes Chagas disease, a major human disease affecting 15 million people and threatening 100 million people in Latin America, and the Salivaria with the anterior station inoculation, mainly African livestock pathogenic trypanosomes, including the agents of sleeping sickness, a major human disease affecting around half a million people and threatening 60 million people in Africa. Now, T. evansi was reported in man is it required to investigate its zoonotic potential?

  12. The effect of Bulgarian propolis against Trypanosoma cruzi and during its interaction with host cells

    Directory of Open Access Journals (Sweden)

    Andréia Pires Dantas

    2006-03-01

    Full Text Available Propolis has shown activity against pathogenic microorganisms that cause diseases in humans and animals. The ethanol (Et-Blg and acetone (Ket-Blg extracts from a Bulgarian propolis, with known chemical compositions, presented similar activity against tissue culture-derived amastigotes. The treatment of Trypanosoma cruzi-infected skeletal muscle cells with Et-Blg led to a decrease of infection and of the intracellular proliferation of amastigotes, while damage to the host cell was observed only at concentration 12.5 times higher than those affecting the parasite. Ultrastructural analysis of the effect of both extracts in epimastigotes revealed that the main targets were the mitochondrion and reservosomes. Et-Blg also affected the mitochondrion-kinetoplast complex in trypomastigotes, offering a potential target for chemotherapeutic agents.

  13. The role of adaptations in two-strain competition for sylvatic Trypanosoma cruzi transmission.

    Science.gov (United States)

    Kribs-Zaleta, Christopher M; Mubayi, Anuj

    2012-01-01

    This study presents a continuous-time model for the sylvatic transmission dynamics of two strains of Trypanosoma cruzi enzootic in North America, in order to study the role that adaptations of each strain to distinct modes of transmission (classical stercorarian transmission on the one hand, and vertical and oral transmission on the other) may play in the competition between the two strains. A deterministic model incorporating contact process saturation predicts competitive exclusion, and reproductive numbers for the infection provide a framework for evaluating the competition in terms of adaptive trade-off between distinct transmission modes. Results highlight the importance of oral transmission in mediating the competition between horizontal (stercorarian) and vertical transmission; its presence as a competing contact process advantages vertical transmission even without adaptation to oral transmission, but such adaptation appears necessary to explain the persistence of (vertically-adapted) T. cruzi IV in raccoons and woodrats in the southeastern United States.

  14. Seroprevalence of Trypanosoma cruzi in rural Ecuador and clustering of seropositivity within households.

    Science.gov (United States)

    Black, Carla L; Ocaña-Mayorga, Sofía; Riner, Diana K; Costales, Jaime A; Lascano, Mauricio S; Arcos-Terán, Laura; Preisser, John S; Seed, J Richard; Grijalva, Mario J

    2009-12-01

    We performed a cross-sectional study of Trypanosoma cruzi seroprevalence in 14 communities in three provinces of Ecuador and estimated the magnitude of the association of seropositive individuals within households. A total of 3,286 subjects from 997 households were included. Seroprevalence was 5.7%, 1.0%, and 3.6% in subjects in the Manabí, Guayas, and Loja provinces, respectively. Seroprevalence increased with increasing age in Manabí and Guayas, whereas in Loja, the highest prevalence occurred in children Loja, the odds of seropositivity were more than two times greater for an individual living in a household with another seropositive person. Our results indicate that transmission of T. cruzi is ongoing in Ecuador, although intensity of transmission and mechanisms of interaction between humans and the insect vectors of disease vary between geographic regions.

  15. The Role of Heme and Reactive Oxygen Species in Proliferation and Survival of Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Marcia Cristina Paes

    2011-01-01

    Full Text Available Trypanosoma cruzi, the protozoan responsible for Chagas disease, has a complex life cycle comprehending two distinct hosts and a series of morphological and functional transformations. Hemoglobin degradation inside the insect vector releases high amounts of heme, and this molecule is known to exert a number of physiological functions. Moreover, the absence of its complete biosynthetic pathway in T. cruzi indicates heme as an essential molecule for this trypanosomatid survival. Within the hosts, T. cruzi has to cope with sudden environmental changes especially in the redox status and heme is able to increase the basal production of reactive oxygen species (ROS which can be also produced as byproducts of the parasite aerobic metabolism. In this regard, ROS sensing is likely to be an important mechanism for the adaptation and interaction of these organisms with their hosts. In this paper we discuss the main features of heme and ROS susceptibility in T. cruzi biology.

  16. Deciphering RNA Regulatory Elements Involved in the Developmental and Environmental Gene Regulation of Trypanosoma brucei.

    Science.gov (United States)

    Gazestani, Vahid H; Salavati, Reza

    2015-01-01

    Trypanosoma brucei is a vector-borne parasite with intricate life cycle that can cause serious diseases in humans and animals. This pathogen relies on fine regulation of gene expression to respond and adapt to variable environments, with implications in transmission and infectivity. However, the involved regulatory elements and their mechanisms of actions are largely unknown. Here, benefiting from a new graph-based approach for finding functional regulatory elements in RNA (GRAFFER), we have predicted 88 new RNA regulatory elements that are potentially involved in the gene regulatory network of T. brucei. We show that many of these newly predicted elements are responsive to both transcriptomic and proteomic changes during the life cycle of the parasite. Moreover, we found that 11 of predicted elements strikingly resemble previously identified regulatory elements for the parasite. Additionally, comparison with previously predicted motifs on T. brucei suggested the superior performance of our approach based on the current limited knowledge of regulatory elements in T. brucei.

  17. Trypanosoma cruzi in the anal glands of urban opossums: I- isolation and experimental infections

    Directory of Open Access Journals (Sweden)

    S Urdaneta-Morales

    1996-08-01

    Full Text Available Opossums (Didelphis marsupialis captured in intensely urbanized areas of the city of Caracas, Venezuela, were found infected with Trypanosoma cruzi. The developmental cycle of trypomastigote-epimastigote-metacyclic infective trypomastigote, usually occurring in the intestine of the triatomine vector, was taking place in the anal odoriferous glands of the opossums. Material from the glands, inoculated in young, healthy opossums and white mice by different routes, subcutaneously, intraperitoneally, orally, and into the eye, induced T. cruzi infections in all animals. Parasitemia, invasion of cardiac and skeletal muscle, and intracellular multiplication of amastigotes were observed. Inoculation of metacyclics from anal glands, cultured in LIT medium, gave equivalent results. All opossums survived; all mice died. Excreta of opossums may thus transmit Chagas' disease by contamination, even in urban areas where insect vectors are not present.

  18. Aspectos ultra-estruturais da forma epimastigota do Trypanosoma cruzi em meio LIT

    Directory of Open Access Journals (Sweden)

    Wanderley de Souza

    1975-06-01

    Full Text Available E feito um estudo da ultra-estrutura da forma epismastigota do Trypanosoma cruzi mantida em meio de cultivo acelular. O núcleo das formas em divisão apresenta um aspecto homogêneo. Microtúbulos intranucleares são observados durante a divisão. No entanto, a membrana nuclear permanece íntegra. O citoplasma apresenta-se com vacúolos de dimensões e aspectos variados. Com o método do ácido periódico-tiosemicarbazida-proteinato de prata, polissacaríáeos e/ou glicoproteínas foram localizados na membrana celular e na membrana que delimita certos vacúolos citoplasmáticos.

  19. Mechanism of Trypanosoma cruzi Placenta Invasion and Infection: The Use of Human Chorionic Villi Explants

    Directory of Open Access Journals (Sweden)

    Ricardo E. Fretes

    2012-01-01

    Full Text Available Congenital Chagas disease, a neglected tropical disease, endemic in Latin America, is associated with premature labor and miscarriage. During vertical transmission the parasite Trypanosoma cruzi (T. cruzi crosses the placental barrier. However, the exact mechanism of the placental infection remains unclear. We review the congenital transmission of T. cruzi, particularly the role of possible local placental factors that contribute to the vertical transmission of the parasite. Additionally, we analyze the different methods available for studying the congenital transmission of the parasite. In that context, the ex vivo infection with T. cruzi trypomastigotes of human placental chorionic villi constitutes an excellent tool for studying parasite infection strategies as well as possible local antiparasitic mechanisms.

  20. First report of surra (Trypanosoma evansi infection in a Tunisian dog

    Directory of Open Access Journals (Sweden)

    Rjeibi Mohamed Ridha

    2015-01-01

    Full Text Available Trypanosoma evansi, the agent of surra, is a salivarian trypanosome, originating from Africa. Surra is a major disease in camels, equines and dogs, in which it can often be fatal in the absence of treatment. Animals exhibit nonspecific clinical signs (anaemia, loss of weight and abortion. In the present survey, a blood sample was collected in Sousse (Central Tunisia from a dog that presented clinical signs of trypanosomiasis. Giemsa-stained blood smears and PCR were performed. ITS1 sequences from blood had 99.8 and 99.5% homology with published T. evansi sequences from cattle and camels, respectively. To our knowledge, this is the first report of T. evansi in a Tunisian dog.

  1. Skin Cancer Screening

    Science.gov (United States)

    ... Genetics of Skin Cancer Skin Cancer Screening Research Skin Cancer Screening (PDQ®)–Patient Version What is screening? ... These are called diagnostic tests . General Information About Skin Cancer Key Points Skin cancer is a disease ...

  2. Screening for Cancer

    Science.gov (United States)

    Cancer screening is checking for cancer in people who don't have symptoms. Screening tests can help doctors find and treat several types of cancer early, but cancer screening can have harms as well as benefits.

  3. Colorectal Cancer Screening

    Science.gov (United States)

    ... Genetics of Colorectal Cancer Colorectal Cancer Screening Research Colorectal Cancer Screening (PDQ®)–Patient Version What is screening? Go ... These are called diagnostic tests . General Information About Colorectal Cancer Key Points Colorectal cancer is a disease in ...

  4. Screen time and children

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/patientinstructions/000355.htm Screen time and children To use the sharing features on ... videos is considered unhealthy screen time. Current Screen Time Guidelines Children under age 2 should have no ...

  5. Stomach (Gastric) Cancer Screening

    Science.gov (United States)

    ... Stomach Cancer Prevention Stomach Cancer Screening Research Stomach (Gastric) Cancer Screening (PDQ®)–Patient Version What is screening? Go ... are called diagnostic tests . General Information About Stomach (Gastric) Cancer Key Points Stomach cancer is a disease in ...

  6. Transcriptional profiling of cattle infected with Trypanosoma congolense highlights gene expression signatures underlying trypanotolerance and trypanosusceptibility

    Directory of Open Access Journals (Sweden)

    Naessens Jan

    2009-05-01

    Full Text Available Abstract Background African animal trypanosomiasis (AAT caused by tsetse fly-transmitted protozoa of the genus Trypanosoma is a major constraint on livestock and agricultural production in Africa and is among the top ten global cattle diseases impacting on the poor. Here we show that a functional genomics approach can be used to identify temporal changes in host peripheral blood mononuclear cell (PBMC gene expression due to disease progression. We also show that major gene expression differences exist between cattle from trypanotolerant and trypanosusceptible breeds. Using bovine long oligonucleotide microarrays and real time quantitative reverse transcription PCR (qRT-PCR validation we analysed PBMC gene expression in naïve trypanotolerant and trypanosusceptible cattle experimentally challenged with Trypanosoma congolense across a 34-day infection time course. Results Trypanotolerant N'Dama cattle displayed a rapid and distinct transcriptional response to infection, with a ten-fold higher number of genes differentially expressed at day 14 post-infection compared to trypanosusceptible Boran cattle. These analyses identified coordinated temporal gene expression changes for both breeds in response to trypanosome infection. In addition, a panel of genes were identified that showed pronounced differences in gene expression between the two breeds, which may underlie the phenomena of trypanotolerance and trypanosusceptibility. Gene ontology (GO analysis demonstrate that the products of these genes may contribute to increased mitochondrial mRNA translational efficiency, a more pronounced B cell response, an elevated activation status and a heightened response to stress in trypanotolerant cattle. Conclusion This study has revealed an extensive and diverse range of cellular processes that are altered temporally in response to trypanosome infection in African cattle. Results indicate that the trypanotolerant N'Dama cattle respond more rapidly and with a

  7. Prevalencia de infeccion a Trypanosoma cruzi en donadores de sangre en el Estado de Jalisco, Mexico

    Directory of Open Access Journals (Sweden)

    Francisco Trujillo Contreras

    1993-06-01

    Full Text Available Durante el periodo de Octubre de 1991 a Marzo de 1992, se tomaron 3419 muestras de donadores de sangre de 12 localidades rurales y de 8 hospitales urbanos a los que se les realizo un estúdio serológico mediante la reacción de hemaglutinación indirecta encontrándose anticuerpos contra Trypanosoma cruzi en 44 indivíduos 39 masculinosy 5 femininos. El 90,9% de donantes fueron masculinos. De acuerdo a su procedencia, el 73,5% fué del área urbana y el 26,5% del área rural. De acuerdo a los resultados el riesgo de transmisión de T. cruzi por transfusión sanguinea está latente por la creciente urbanización de la enfermedad de Chagas.A Chagas Disease serological study was done frorn October 1991 to March 1992 and 3419 samples were takenfrom people who donated blood at 12 county areas of Jalisco, México and 8 urban hospitais, by means of indirect hemagglutination reaction. The results indicate that: 73.5% of the donors were from urban area, 26.5% were from rural areas; 1.28% of the donors (N=44 were considered infected. Thirty nine of them (1.14 were males and 5 females. According to the above mentioned data, we can confirm that the risk of transmission of Trypanosoma cruzi can occur by blood transfusion and this is potentially latent because of the growing urbanization of Chagas disease.

  8. Interactions between 4-aminoquinoline and heme: Promising mechanism against Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Guilherme Curty Lechuga

    2016-12-01

    Full Text Available Chagas disease is a neglected tropical disease caused by the flagellated protozoan Trypanosoma cruzi. The current drugs used to treat this disease have limited efficacy and produce severe side effects. Quinolines, nitrogen heterocycle compounds that form complexes with heme, have a broad spectrum of antiprotozoal activity and are a promising class of new compounds for Chagas disease chemotherapy. In this study, we evaluated the activity of a series of 4-arylaminoquinoline-3-carbonitrile derivatives against all forms of Trypanosoma cruzi in vitro. Compound 1g showed promising activity against epimastigote forms when combined with hemin (IC50<1 μM, with better performance than benznidazole, the reference drug. This compound also inhibited the viability of trypomastigotes and intracellular amastigotes. The potency of 1g in combination with heme was enhanced against epimastigotes and trypomastigotes, suggesting a similar mechanism of action that occurs in Plasmodium spp. The addition of hemin to the culture medium increased trypanocidal activity of analog 1g without changing the cytotoxicity of the host cell, reaching an IC50 of 11.7 μM for trypomastigotes. The mechanism of action was demonstrated by the interaction of compound 1g with hemin in solution and prevention of heme peroxidation. Compound 1g and heme treatment induced alterations of the mitochondrion-kinetoplast complex in epimastigotes and trypomastigotes and also, accumulation of electron-dense deposits in amastigotes as visualized by transmission electron microscopy. The trypanocidal activity of 4-aminoquinolines and the elucidation of the mechanism involving interaction with heme is a neglected field of research, given the parasite's lack of heme biosynthetic pathway and the importance of this cofactor for parasite survival and growth. The results of this study can improve and guide rational drug development and combination treatment strategies.

  9. Trypanosoma cruzi Detection in Colombian Patients with a Diagnosis of Esophageal Achalasia.

    Science.gov (United States)

    Panesso-Gómez, Santiago; Pavia, Paula; Rodríguez-Mantilla, Iván Enrique; Lasso, Paola; Orozco, Luis A; Cuellar, Adriana; Puerta, Concepción J; Mendoza de Molano, Belén; González, John M

    2018-03-01

    Achalasia is a motility disorder of the esophagus that might be secondary to a chronic Trypanosoma cruzi infection. Several studies have investigated esophageal achalasia in patients with Chagas disease (CD) in Latin America, but no related studies have been performed in Colombia. The goals of the present study were to determine the presence of anti- T. cruzi antibodies in patients with esophageal achalasia who visited a referral hospital in Bogotá, Colombia, and to detect the presence of the parasite and its discrete typing units (DTUs). This cross-sectional study was conducted in adult patients (18-65 years old) who were previously diagnosed with esophageal achalasia and from whom blood was drawn to assess antibodies against T. cruzi using four different serological tests. Trypanosoma cruzi DNA was detected by conventional polymerase chain reaction (cPCR) and quantitative polymerase chain reaction (qPCR). In total, 38 patients, with an average age of 46.6 years (standard deviation of ±16.2) and comprising 16 men and 22 women, were enrolled. Five (13.15%) patients were found to be positive for anti- T. cruzi antibodies by indirect immunofluorescence assay (IFA), and two patients who were negative according to IFA were reactive by both enzyme-linked immunosorbent assay and immunoblot (5.3%). Parasite DNA was detected in two of these seven patients by cPCR and in one of these by qPCR. The parasite DTU obtained was TcI. In summary, this study identified T. cruzi in Colombian patients with esophageal achalasia, indicating that digestive compromise could also be present in patients with chronic CD.

  10. Interactions between 4-aminoquinoline and heme: Promising mechanism against Trypanosoma cruzi.

    Science.gov (United States)

    Lechuga, Guilherme Curty; Borges, Júlio Cesar; Calvet, Claudia Magalhães; de Araújo, Humberto Pinheiro; Zuma, Aline Araujo; do Nascimento, Samara Braga; Motta, Maria Cristina Machado; Bernardino, Alice Maria Rolim; Pereira, Mirian Claudia de Souza; Bourguignon, Saulo Cabral

    2016-12-01

    Chagas disease is a neglected tropical disease caused by the flagellated protozoan Trypanosoma cruzi. The current drugs used to treat this disease have limited efficacy and produce severe side effects. Quinolines, nitrogen heterocycle compounds that form complexes with heme, have a broad spectrum of antiprotozoal activity and are a promising class of new compounds for Chagas disease chemotherapy. In this study, we evaluated the activity of a series of 4-arylaminoquinoline-3-carbonitrile derivatives against all forms of Trypanosoma cruzi in vitro. Compound 1g showed promising activity against epimastigote forms when combined with hemin (IC50<1 μM), with better performance than benznidazole, the reference drug. This compound also inhibited the viability of trypomastigotes and intracellular amastigotes. The potency of 1g in combination with heme was enhanced against epimastigotes and trypomastigotes, suggesting a similar mechanism of action that occurs in Plasmodium spp. The addition of hemin to the culture medium increased trypanocidal activity of analog 1g without changing the cytotoxicity of the host cell, reaching an IC50 of 11.7 μM for trypomastigotes. The mechanism of action was demonstrated by the interaction of compound 1g with hemin in solution and prevention of heme peroxidation. Compound 1g and heme treatment induced alterations of the mitochondrion-kinetoplast complex in epimastigotes and trypomastigotes and also, accumulation of electron-dense deposits in amastigotes as visualized by transmission electron microscopy. The trypanocidal activity of 4-aminoquinolines and the elucidation of the mechanism involving interaction with heme is a neglected field of research, given the parasite's lack of heme biosynthetic pathway and the importance of this cofactor for parasite survival and growth. The results of this study can improve and guide rational drug development and combination treatment strategies. Copyright © 2016 The Authors. Published by Elsevier

  11. The Effectiveness of Natural Diarylheptanoids against Trypanosoma cruzi: Cytotoxicity, Ultrastructural Alterations and Molecular Modeling Studies.

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    Vitor Sueth-Santiago

    Full Text Available Curcumin (CUR is the major constituent of the rhizomes of Curcuma longa and has been widely investigated for its chemotherapeutic properties. The well-known activity of CUR against Leishmania sp., Trypanosoma brucei and Plasmodium falciparum led us to investigate its activity against Trypanosoma cruzi. In this work, we tested the cytotoxic effects of CUR and other natural curcuminoids on different forms of T. cruzi, as well as the ultrastructural changes induced in epimastigote form of the parasite. CUR was verified as the curcuminoid with more significant trypanocidal properties (IC50 10.13 μM on epimastigotes. Demethoxycurcumin (DMC was equipotent to CUR (IC50 11.07 μM, but bisdemethoxycurcumin (BDMC was less active (IC50 45.33 μM and cyclocurcumin (CC was inactive. In the experiment with infected murine peritoneal macrophages all diarylheptanoids were more active than the control in the inhibition of the trypomastigotes release. The electron microscopy images showed ultrastructural changes associated with the cytoskeleton of the parasite, indicating tubulin as possible target of CUR in T. cruzi. The results obtained by flow cytometry analysis of DNA content of the parasites treated with natural curcuminoids suggested a mechanism of action on microtubules related to the paclitaxel`s mode of action. To better understand the mechanism of action highlighted by electron microscopy and flow cytometry experiments we performed the molecular docking of natural curcuminoids on tubulin of T. cruzi in a homology model and the results obtained showed that the observed interactions are in accordance with the IC50 values found, since there CUR and DMC perform similar interactions at the binding site on tubulin while BDMC do not realize a hydrogen bond with Lys163 residue due to the absence of methoxyl groups. These results indicate that trypanocidal properties of CUR may be related to the cytoskeletal alterations.

  12. Population genetics of Trypanosoma brucei rhodesiense: clonality and diversity within and between foci.

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    Craig W Duffy

    2013-11-01

    Full Text Available African trypanosomes are unusual among pathogenic protozoa in that they can undergo their complete morphological life cycle in the tsetse fly vector with mating as a non-obligatory part of this development. Trypanosoma brucei rhodesiense, which infects humans and livestock in East and Southern Africa, has classically been described as a host-range variant of the non-human infective Trypanosoma brucei that occurs as stable clonal lineages. We have examined T. b. rhodesiense populations from East (Uganda and Southern (Malawi Africa using a panel of microsatellite markers, incorporating both spatial and temporal analyses. Our data demonstrate that Ugandan T. b. rhodesiense existed as clonal populations, with a small number of highly related genotypes and substantial linkage disequilibrium between pairs of loci. However, these populations were not stable as the dominant genotypes changed and the genetic diversity also reduced over time. Thus these populations do not conform to one of the criteria for strict clonality, namely stability of predominant genotypes over time, and our results show that, in a period in the mid 1990s, the previously predominant genotypes were not detected but were replaced by a novel clonal population with limited genetic relationship to the original population present between 1970 and 1990. In contrast, the Malawi T. b. rhodesiense population demonstrated significantly greater diversity and evidence for frequent genetic exchange. Therefore, the population genetics of T. b. rhodesiense is more complex than previously described. This has important implications for the spread of the single copy T. b. rhodesiense gene that allows human infectivity, and therefore the epidemiology of the human disease, as well as suggesting that these parasites represent an important organism to study the influence of optional recombination upon population genetic dynamics.

  13. Heme A synthesis and CcO activity are essential for Trypanosoma cruzi infectivity and replication.

    Science.gov (United States)

    Merli, Marcelo L; Cirulli, Brenda A; Menéndez-Bravo, Simón M; Cricco, Julia A

    2017-06-27

    Trypanosoma cruzi , the causative agent of Chagas disease, presents a complex life cycle and adapts its metabolism to nutrients' availability. Although T. cruzi is an aerobic organism, it does not produce heme. This cofactor is acquired from the host and is distributed and inserted into different heme-proteins such as respiratory complexes in the parasite's mitochondrion. It has been proposed that T. cruzi's energy metabolism relies on a branched respiratory chain with a cytochrome c oxidase-type aa 3 (C c O) as the main terminal oxidase. Heme A, the cofactor for all eukaryotic C c O, is synthesized via two sequential enzymatic reactions catalyzed by heme O synthase (HOS) and heme A synthase (HAS). Previously, TcCox10 and TcCox15 ( Trypanosoma cruzi Cox10 and Cox15 proteins) were identified in T. cruzi They presented HOS and HAS activity, respectively, when they were expressed in yeast. Here, we present the first characterization of TcCox15 in T. cruzi , confirming its role as HAS. It was differentially detected in the different T. cruzi stages, being more abundant in the replicative forms. This regulation could reflect the necessity of more heme A synthesis, and therefore more C c O activity at the replicative stages. Overexpression of a non-functional mutant caused a reduction in heme A content. Moreover, our results clearly showed that this hindrance in the heme A synthesis provoked a reduction on C c O activity and, in consequence, an impairment on T. cruzi survival, proliferation and infectivity. This evidence supports that T. cruzi depends on the respiratory chain activity along its life cycle, being C c O an essential terminal oxidase. © 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

  14. Chemical characterisation of Nigerian red propolis and its biological activity against Trypanosoma Brucei.

    Science.gov (United States)

    Omar, Ruwida M K; Igoli, John; Gray, Alexander I; Ebiloma, Godwin Unekwuojo; Clements, Carol; Fearnley, James; Ebel, Ru Angeli Edrada; Zhang, Tong; De Koning, Harry P; Watson, David G

    2016-01-01

    A previous study showed the unique character of Nigerian red propolis from Rivers State, Nigeria (RSN), with regards to chemical composition and activity against Trypanosoma brucei in comparison with other African propolis. To carry out fractionation and biological testing of Nigerian propolis in order to isolate compounds with anti-trypanosomal activity. To compare the composition of the RSN propolis with the composition of Brazilian red propolis. Profiling was carried out using HPLC-UV-ELSD and HPLC-Orbitrap-FTMS on extracts of two samples collected from RSN with data extraction using MZmine software. Isolation was carried out by normal phase and reversed phase MPLC. Elucidation of the compounds with a purity > 95% was performed by 1D/2D NMR HRMS and HRLC-MS(n) . Ten phenolic compounds were isolated or in the case of liquiritigenin partially purified. Data for nine of these correlated with literature reports of known compounds i.e. one isoflavanone, calycosin (1); two flavanones, liquiritigenin (2) and pinocembrin (5); an isoflavan, vestitol (3); a pterocarpan, medicarpin (4); two prenylflavanones, 8-prenylnaringenin (7) and 6-prenylnaringenin (8); and two geranyl flavonoids, propolin D (9) and macarangin (10). The tenth was elucidated as a previously undescribed dihydrobenzofuran (6). The isolated compounds were tested against Trypanosoma brucei and displayed moderate to high activity. Some of the compounds tested had similar activity against wild type T. brucei and two strains displaying pentamidine resistance. Nigerian propolis from RSN has some similarities with Brazilian red propolis. The propolis displayed anti-trypanosomal activity at a potentially useful level. Copyright © 2015 John Wiley & Sons, Ltd.

  15. Congenital transmission of Trypanosoma cruzi in Argentina, Honduras, and Mexico: study protocol

    Science.gov (United States)

    2013-01-01

    Background Trypanosoma cruzi has been divided into Discrete Typing Units I and non-I (II-VI). T. cruzi I is predominant in Mexico and Central America, while non-I is predominant in most of South America, including Argentina. Little is known about congenital transmission of T. cruzi I. The specific aim of this study is to determine the rate of congenital transmission of T. cruzi I compared to non-I. Methods/design We are conducting a prospective study to enroll at delivery, 10,000 women in Argentina, 7,500 women in Honduras, and 13,000 women in Mexico. We are measuring transmitted maternal T. cruzi antibodies by performing two rapid tests in cord blood (Stat-Pak, Chembio, Medford, New York, and Trypanosoma Detect, InBios, Seattle, Washington). If at least one of the results is positive, we are identifying infants who are congenitally infected by performing parasitological examinations on cord blood and at 4–8 weeks, and serological follow-up at 10 months. Serological confirmation by ELISA (Wiener, Rosario, Argentina) is performed in cord and maternal blood, and at 10 months. We also are performing T. cruzi standard PCR, real-time quantitative PCR and genotyping on maternal venous blood and on cord blood, and serological examinations on siblings. Data are managed by a Data Center in Montevideo, Uruguay. Data are entered online at the sites in an OpenClinica data management system, and digital pictures of data forms are sent to the Data Center for quality control. Weekly reports allow for rapid feedback to the sites. Trial registration Observational study with ClinicalTrials.gov Identifier NCT01787968 PMID:24119247

  16. Stearoyl-CoA desaturase is an essential enzyme for the parasitic protist Trypanosoma brucei

    International Nuclear Information System (INIS)

    Alloatti, Andres; Gupta, Shreedhara; Gualdron-Lopez, Melisa; Nguewa, Paul A.; Altabe, Silvia G.; Deumer, Gladys; Wallemacq, Pierre; Michels, Paul A.M.; Uttaro, Antonio D.

    2011-01-01

    Highlights: → Inhibiting Δ9 desaturase drastically changes T. brucei's fatty-acid composition. → Isoxyl specifically inhibits the Δ9 desaturase causing a growth arrest. → RNA interference of desaturase expression causes a similar effect. → Feeding T. brucei-infected mice with Isoxyl decreases the parasitemia. → 70% of Isoxyl-treated mice survived the trypanosome infection. -- Abstract: Trypanosoma brucei, the etiologic agent of sleeping sickness, is exposed to important changes in nutrients and temperature during its life cycle. To adapt to these changes, the fluidity of its membranes plays a crucial role. This fluidity, mediated by the fatty-acid composition, is regulated by enzymes named desaturases. We have previously shown that the oleoyl desaturase is essential for Trypanosoma cruzi and T. brucei. In this work, we present experimental support for the relevance of stearoyl-CoA desaturase (SCD) for T. brucei's survival, in both its insect or procyclic-form (PCF) and bloodstream-form (BSF) stages. We evaluated this essentiality in two different ways: by generating a SCD knocked-down parasite line using RNA interference, and by chemical inhibition of the enzyme with two compounds, Isoxyl and a thiastearate with the sulfur atom at position 10 (10-TS). The effective concentration for 50% growth inhibition (EC 50 ) of PCF was 1.0 ± 0.2 μM for Isoxyl and 5 ± 2 μM for 10-TS, whereas BSF appeared more susceptible with EC 50 values 0.10 ± 0.03 μM (Isoxyl) and 1.0 ± 0.6 μM (10-TS). RNA interference showed to be deleterious for both stages of the parasite. In addition, T. brucei-infected mice were fed with Isoxyl, causing a reduction of the parasitemia and an increase of the rodents' survival.

  17. Trypanosoma Infection Favors Brucella Elimination via IL-12/IFNγ-Dependent Pathways

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    Arnaud Machelart

    2017-07-01

    Full Text Available This study develops an original co-infection model in mice using Brucella melitensis, the most frequent cause of human brucellosis, and Trypanosoma brucei, the agent of African trypanosomiasis. Although the immunosuppressive effects of T. brucei in natural hosts and mice models are well established, we observed that the injection of T. brucei in mice chronically infected with B. melitensis induces a drastic reduction in the number of B. melitensis in the spleen, the main reservoir of the infection. Similar results are obtained with Brucella abortus- and Brucella suis-infected mice and B. melitensis-infected mice co-infected with Trypanosoma cruzi, demonstrating that this phenomenon is not due to antigenic cross-reactivity. Comparison of co-infected wild-type and genetically deficient mice showed that Brucella elimination required functional IL-12p35/IFNγ signaling pathways and the presence of CD4+ T cells. However, the impact of wild type and an attenuated mutant of T. brucei on B. melitensis were similar, suggesting that a chronic intense inflammatory reaction is not required to eliminate B. melitensis. Finally, we also tested the impact of T. brucei infection on the course of Mycobacterium tuberculosis infection. Although T. brucei strongly increases the frequency of IFNγ+CD4+ T cells, it does not ameliorate the control of M. tuberculosis infection, suggesting that it is not controlled by the same effector mechanisms as Brucella. Thus, whereas T. brucei infections are commonly viewed as immunosuppressive and pathogenic, our data suggest that these parasites can specifically affect the immune control of Brucella infection, with benefits for the host.

  18. High Trypanosoma cruzi infection prevalence associated with minimal cardiac pathology among wild carnivores in central Texas

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    Rachel Curtis-Robles

    2016-08-01

    Full Text Available Infection with the zoonotic vector-borne protozoal parasite Trypanosoma cruzi causes Chagas disease in humans and dogs throughout the Americas. Despite the recognized importance of various wildlife species for perpetuating Trypanosoma cruzi in nature, relatively little is known about the development of cardiac disease in infected wildlife. Using a cross-sectional study design, we collected cardiac tissue and blood from hunter-donated wildlife carcasses- including raccoon (Procyon lotor, coyote (Canis latrans, gray fox (Urocyon cinereoargenteus, and bobcat (Lynx rufus – from central Texas, a region with established populations of infected triatomine vectors and increasing diagnoses of Chagas disease in domestic dogs. Based on PCR analysis, we found that 2 bobcats (14.3%, 12 coyotes (14.3%, 8 foxes (13.8%, and 49 raccoons (70.0% were positive for T. cruzi in at least one sample (right ventricle, apex, and/or blood clot. Although a histologic survey of right ventricles showed that 21.1% of 19 PCR-positive hearts were characterized by mild lymphoplasmocytic infiltration, no other lesions and no amastigotes were observed in any histologic section. DNA sequencing of the TcSC5D gene revealed that raccoons were infected with T. cruzi strain TcIV, and a single racoon harbored a TcI/TcIV mixed infection. Relative to other wildlife species tested here, our data suggest that raccoons may be important reservoirs of TcIV in Texas and a source of infection for indigenous triatomine bugs. The overall high level of infection in this wildlife community likely reflects high levels of vector contact, including ingestion of bugs. Although the relationship between the sylvatic cycle of T. cruzi transmission and human disease risk in the United States has yet to be defined, our data suggest that hunters and wildlife professionals should take precautions to avoid direct contact with potentially infected wildlife tissues.

  19. Epidemiology of Babesia, Anaplasma and Trypanosoma species using a new expanded reverse line blot hybridization assay.

    Science.gov (United States)

    Paoletta, Martina Soledad; López Arias, Ludmila; de la Fournière, Sofía; Guillemi, Eliana Carolina; Luciani, Carlos; Sarmiento, Néstor Fabián; Mosqueda, Juan; Farber, Marisa Diana; Wilkowsky, Silvina Elizabeth

    2018-02-01

    Vector-borne hemoparasitic infections are a major problem that affects livestock industries worldwide, particularly in tropical and subtropical regions. In this work, a reverse line blot (RLB) hybridization assay was developed for the simultaneous detection and identification of Anaplasma, Babesia and bovine trypanosomes, encompassing in this way the most relevant hemoparasites that affect cattle. A total of 186 bovine blood samples collected from two different ecoepidemiological regions of northeast Argentina, with and without tick control, were analyzed with this new RLB. High diversity of parasites, such as Babesia bovis, B. bigemina, Anaplasma marginale and three different Trypanosoma species, was found. High rates of coinfections were also detected, and significant differences were observed not only in the prevalence of parasites but also in the level of coinfections between the two analyzed areas. Regarding the Trypanosoma genus, we provide molecular evidence of the presence of T. vivax and T. theileri for the first time in Argentina. Besides, since the RLB is a prospective tool, it allowed the identification of a yet unknown bovine trypanosome which could not be assigned to any of the bovine species known so far. In the present study we provide new insights on the prevalence of several pathogens that directly impact on livestock production in Argentina. The RLB assay developed here allows to identify simultaneously numerous pathogenic species which can also be easily expanded to detect other blood borne pathogens. These characteristics make the RLB hybridization assay an essential tool for epidemiological survey of all vector-borne pathogens. Copyright © 2017 Elsevier GmbH. All rights reserved.

  20. Projected future distributions of vectors of Trypanosoma cruzi in North America under climate change scenarios.

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    Miroslava Garza

    2014-05-01

    Full Text Available Chagas disease kills approximately 45 thousand people annually and affects 10 million people in Latin America and the southern United States. The parasite that causes the disease, Trypanosoma cruzi, can be transmitted by insects of the family Reduviidae, subfamily Triatominae. Any study that attempts to evaluate risk for Chagas disease must focus on the ecology and biogeography of these vectors. Expected distributional shifts of vector species due to climate change are likely to alter spatial patterns of risk of Chagas disease, presumably through northward expansion of high risk areas in North America.We forecast the future (2050 distributions in North America of Triatoma gerstaeckeri and T. sanguisuga, two of the most common triatomine species and important vectors of Trypanosoma cruzi in the southern United States. Our aim was to analyze how climate change might affect the future shift of Chagas disease in North America using a maximum entropy algorithm to predict changes in suitable habitat based on vector occurrence points and predictive environmental variables. Projections based on three different general circulation models (CCCMA, CSIRO, and HADCM3 and two IPCC scenarios (A2 and B2 were analyzed. Twenty models were developed for each case and evaluated via cross-validation. The final model averages result from all twenty of these models. All models had AUC >0.90, which indicates that the models are robust. Our results predict a potential northern shift in the distribution of T. gerstaeckeri and a northern and southern distributional shift of T. sanguisuga from its current range due to climate change.The results of this study provide baseline information for monitoring the northward shift of potential risk from Chagas disease in the face of climate change.

  1. Nitrogen rate and plant population effects on yield and yield ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-12-17

    Gan et al., 2003). Nitrogen increases yield by influencing a variety of agronomic and quality parameters. In general, there was an increase in plant height and dry matter accumulation per plant in soybean (Manral and Saxena, ...

  2. Nitrogen rate and plant population effects on yield and yield ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-12-17

    Dec 17, 2008 ... density and nitrogen rate increased plant height, lowest pod height, harvest index and seed yield. ... since some combine harvester heads are unable to pick ..... as effected by population density and plant distribution.

  3. Trypanosoma janseni n. sp. (Trypanosomatida: Trypanosomatidae isolated from Didelphis aurita (Mammalia: Didelphidae in the Atlantic Rainforest of Rio de Janeiro, Brazil: integrative taxonomy and phylogeography within the Trypanosoma cruzi clade

    Directory of Open Access Journals (Sweden)

    Camila Madeira Tavares Lopes

    Full Text Available BACKGROUND Didelphis spp. are a South American marsupial species that are among the most ancient hosts for the Trypanosoma spp. OBJECTIVES We characterise a new species (Trypanosoma janseni n. sp. isolated from the spleen and liver tissues of Didelphis aurita in the Atlantic Rainforest of Rio de Janeiro, Brazil. METHODS The parasites were isolated and a growth curve was performed in NNN and Schneider's media containing 10% foetal bovine serum. Parasite morphology was evaluated via light microscopy on Giemsa-stained culture smears, as well as scanning and transmission electron microscopy. Molecular taxonomy was based on a partial region (737-bp of the small subunit (18S ribosomal RNA gene and 708 bp of the nuclear marker, glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH genes. Maximum likelihood and Bayesian inference methods were used to perform a species coalescent analysis and to generate individual and concatenated gene trees. Divergence times among species that belong to the T. cruzi clade were also inferred. FINDINGS In vitro growth curves demonstrated a very short log phase, achieving a maximum growth rate at day 3 followed by a sharp decline. Only epimastigote forms were observed under light and scanning microscopy. Transmission electron microscopy analysis showed structures typical to Trypanosoma spp., except one structure that presented as single-membraned, usually grouped in stacks of three or four. Phylogeography analyses confirmed the distinct species status of T. janseni n. sp. within the T. cruzi clade. Trypanosoma janseni n. sp. clusters with T. wauwau in a well-supported clade, which is exclusive and monophyletic. The separation of the South American T. wauwau + T. janseni coincides with the separation of the Southern Super Continent. CONCLUSIONS This clade is a sister group of the trypanosomes found in Australian marsupials and its discovery sheds light on the initial diversification process based on what we currently

  4. Trypanosoma janseni n. sp. (Trypanosomatida: Trypanosomatidae) isolated from Didelphis aurita (Mammalia: Didelphidae) in the Atlantic Rainforest of Rio de Janeiro, Brazil: integrative taxonomy and phylogeography within the Trypanosoma cruzi clade.

    Science.gov (United States)

    Lopes, Camila Madeira Tavares; Menna-Barreto, Rubem Figueiredo Sadok; Pavan, Márcio Galvão; Pereira, Mirian Cláudia De Souza; Roque, André Luiz R

    2018-01-01

    Didelphis spp. are a South American marsupial species that are among the most ancient hosts for the Trypanosoma spp. We characterise a new species (Trypanosoma janseni n. sp.) isolated from the spleen and liver tissues of Didelphis aurita in the Atlantic Rainforest of Rio de Janeiro, Brazil. The parasites were isolated and a growth curve was performed in NNN and Schneider's media containing 10% foetal bovine serum. Parasite morphology was evaluated via light microscopy on Giemsa-stained culture smears, as well as scanning and transmission electron microscopy. Molecular taxonomy was based on a partial region (737-bp) of the small subunit (18S) ribosomal RNA gene and 708 bp of the nuclear marker, glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) genes. Maximum likelihood and Bayesian inference methods were used to perform a species coalescent analysis and to generate individual and concatenated gene trees. Divergence times among species that belong to the T. cruzi clade were also inferred. In vitro growth curves demonstrated a very short log phase, achieving a maximum growth rate at day 3 followed by a sharp decline. Only epimastigote forms were observed under light and scanning microscopy. Transmission electron microscopy analysis showed structures typical to Trypanosoma spp., except one structure that presented as single-membraned, usually grouped in stacks of three or four. Phylogeography analyses confirmed the distinct species status of T. janseni n. sp. within the T. cruzi clade. Trypanosoma janseni n. sp. clusters with T. wauwau in a well-supported clade, which is exclusive and monophyletic. The separation of the South American T. wauwau + T. janseni coincides with the separation of the Southern Super Continent. This clade is a sister group of the trypanosomes found in Australian marsupials and its discovery sheds light on the initial diversification process based on what we currently know about the T. cruzi clade.

  5. Trypanosoma cruzi prevalence and clinical forms in blood donor candidates in Brazil Prevalência e formas clínicas de Trypanosoma cruzi em candidatos a doadores de sangue no Brasil

    Directory of Open Access Journals (Sweden)

    H J Silveira

    2003-12-01

    Full Text Available The prevalence and clinical forms of Trypanosoma cruzi were evaluated among blood donor candidates attended at a general hospital in Rio de Janeiro, Brazil, from January 1997 to April 1999. The investigation was done by means of the indirect hemagglutination test and was confirmed via ELISA. Data were collected from clinical examinations, conventional electrocardiogram, chest radiography and echocar-diography. The results showed that despite Trypanosoma cruzi prevalence of 1.17% (128 patients, mainly in males aged 40 years or over, 70.8% of these patients, mainly males aged 19 to 39 years, demonstrated abnormalities that allowed the diagnosis of cardiopathy and/or esophagopathy. This once again corroborates the importance of Trypanosoma cruzi infection in urban centers.A prevalência e a manifestação das formas clinicas de Trypanosoma cruzi foram avaliadas em candidatos a doadores de sangue atendidos em um hospital geral de Nova Iguaçu, Rio de Janeiro, Brasil, no período de janeiro de 1997 a abril de 1999. A pesquisa sorológica foi realizada por meio do teste de hemaglutinação indireta e confirmada pelo ELISA. Os dados foram coletados considerando os exames clínicos, eletrocardiograma convencional, radiografia de tórax e ecocardiografia. Os resultados demonstraram que, apesar da prevalência ser de 1,17% (128 pacientes, principalmente entre homens com idade igual ou superior a 40 anos, 70,8%, principalmente de homens entre 19 e 39 anos, demonstraram alterações que permitiram o diagnóstico de cardiopatias e/ou esofagopatias, ratificando mais uma vez sua importância nos centros urbanos.

  6. Effects of application boron on yields, yield component and oil ...

    African Journals Online (AJOL)

    The study was conducted to investigate the effects of five boron (B) doses; 0, 2.5, 5.0, 7.5 and 10.0 kg B ha-1 in B-deficient calcareous soils on yield and some yield components of four sunflower genotypes. Genotypes have shown variations with respect to their responses to B applications. AS-615 and Coban had the ...

  7. Decomposing global crop yield variability

    Science.gov (United States)

    Ben-Ari, Tamara; Makowski, David

    2014-11-01

    Recent food crises have highlighted the need to better understand the between-year variability of agricultural production. Although increasing future production seems necessary, the globalization of commodity markets suggests that the food system would also benefit from enhanced supplies stability through a reduction in the year-to-year variability. Here, we develop an analytical expression decomposing global crop yield interannual variability into three informative components that quantify how evenly are croplands distributed in the world, the proportion of cultivated areas allocated to regions of above or below average variability and the covariation between yields in distinct world regions. This decomposition is used to identify drivers of interannual yield variations for four major crops (i.e., maize, rice, soybean and wheat) over the period 1961-2012. We show that maize production is fairly spread but marked by one prominent region with high levels of crop yield interannual variability (which encompasses the North American corn belt in the USA, and Canada). In contrast, global rice yields have a small variability because, although spatially concentrated, much of the production is located in regions of below-average variability (i.e., South, Eastern and South Eastern Asia). Because of these contrasted land use allocations, an even cultivated land distribution across regions would reduce global maize yield variance, but increase the variance of global yield rice. Intermediate results are obtained for soybean and wheat for which croplands are mainly located in regions with close-to-average variability. At the scale of large world regions, we find that covariances of regional yields have a negligible contribution to global yield variance. The proposed decomposition could be applied at any spatial and time scales, including the yearly time step. By addressing global crop production stability (or lack thereof) our results contribute to the understanding of a key

  8. Evaluation of the immune response to CRA and FRA recombinant antigens of Trypanosoma cruzi in C57BL/6 mice.

    Science.gov (United States)

    Pereira, Valéria Rêgo Alves; de Lorena, Virginia Maria Barros; Nakazawa, Mineo; da Silva, Ana Paula Galvão; Montarroyos, Ulisses; Correa-Oliveira, Rodrigo; Gomes, Yara de Miranda

    2003-01-01

    Humoral and cellular immune responses were evaluated in 44 C57BL/6 mice immunized with the Trypanosoma cruzi recombinant antigens CRA and FRA. Both antigens induced cutaneous immediate-type hypersensitivity response. The levels of IgG1, IgG2a, IgG2b and IgG3 were high in CRA immunized mice. IgG3 was the predominant isotype. Although no difference in antibody levels was observed in FRA-immunized mice when compared to control mice, both antigens were able to induce lymphoproliferation in immunized mice. Significant differences were observed between incorporation of [ H]- thymidine by spleen cell stimulated in vitro with CRA or FRA and the control group. These results suggest that CRA and FRA could be involved in mechanisms of resistance to Trypanosoma cruzi infection.

  9. Prenatal screening and genetics

    DEFF Research Database (Denmark)

    Alderson, P; Aro, A R; Dragonas, T

    2001-01-01

    Although the term 'genetic screening' has been used for decades, this paper discusses how, in its most precise meaning, genetic screening has not yet been widely introduced. 'Prenatal screening' is often confused with 'genetic screening'. As we show, these terms have different meanings, and we...... examine definitions of the relevant concepts in order to illustrate this point. The concepts are i) prenatal, ii) genetic screening, iii) screening, scanning and testing, iv) maternal and foetal tests, v) test techniques and vi) genetic conditions. So far, prenatal screening has little connection...... with precisely defined genetics. There are benefits but also disadvantages in overstating current links between them in the term genetic screening. Policy making and professional and public understandings about screening could be clarified if the distinct meanings of prenatal screening and genetic screening were...

  10. screening of pleurotus ostreatus and gleophylum sepiarium strains ...

    African Journals Online (AJOL)

    USER

    sawdust dump-site were screened for protease enzyme production. High yields of protease ... glass spreader, sterilized by dipping in 95% ethanol and flaming. .... because higher temperatures led to a sharp decrease in activity. The result is ...

  11. Sensitivity and Specificity of a Prototype Rapid Diagnostic Test for the Detection of Trypanosoma brucei gambiense Infection: A Multi-centric Prospective Study.

    Science.gov (United States)

    Bisser, Sylvie; Lumbala, Crispin; Nguertoum, Etienne; Kande, Victor; Flevaud, Laurence; Vatunga, Gedeao; Boelaert, Marleen; Büscher, Philippe; Josenando, Theophile; Bessell, Paul R; Biéler, Sylvain; Ndung'u, Joseph M

    2016-04-01

    A major challenge in the control of human African trypanosomiasis (HAT) is lack of reliable diagnostic tests that are rapid and easy to use in remote areas where the disease occurs. In Trypanosoma brucei gambiense HAT, the Card Agglutination Test for Trypanosomiasis (CATT) has been the reference screening test since 1978, usually on whole blood, but also in a 1/8 dilution (CATT 1/8) to enhance specificity. However, the CATT is not available in a single format, requires a cold chain for storage, and uses equipment that requires electricity. A solution to these challenges has been provided by rapid diagnostic tests (RDT), which have recently become available. A prototype immunochromatographic test, the SD BIOLINE HAT, based on two native trypanosomal antigens (VSG LiTat 1.3 and VSG LiTat 1.5) has been developed. We carried out a non-inferiority study comparing this prototype to the CATT 1/8 in field settings. The prototype SD BIOLINE HAT, the CATT Whole Blood and CATT 1/8 were systematically applied on fresh blood samples obtained from 14,818 subjects, who were prospectively enrolled through active and passive screening in clinical studies in three endemic countries of central Africa: Angola, the Democratic Republic of the Congo and the Central African Republic. One hundred and forty nine HAT cases were confirmed by parasitology. The sensitivity and specificity of the prototype SD BIOLINE HAT was 89.26% (95% confidence interval (CI) = 83.27-93.28) and 94.58% (95% CI = 94.20-94.94) respectively. The sensitivity and specificity of the CATT on whole blood were 93.96% (95% CI = 88.92-96.79) and 95.91% (95% CI = 95.58-96.22), and of the CATT 1/8 were 89.26% (95% CI = 83.27-93.28) and 98.88% (95% CI = 98.70-99.04) respectively. After further optimization, the prototype SD BIOLINE HAT could become an alternative to current screening methods in primary healthcare settings in remote, resource-limited regions where HAT typically occurs.

  12. Acute Trypanosoma cruzi Infection in Mouse Induces Infertility or Placental Parasite Invasion and Ischemic Necrosis Associated with Massive Fetal Loss

    OpenAIRE

    Mjihdi, Abdelkarim; Lambot, Marie-Alexandra; Stewart, Ian J.; Detournay, Olivier; Noël, Jean-Christophe; Carlier, Yves; Truyens, Carine

    2002-01-01

    Pathogens may impair reproduction in association or not with congenital infections. We have investigated the effect of acute infection with Trypanosoma cruzi, the protozoan agent of Chagas’ disease in Latin America, on reproduction of mice. Although mating of infected mice occurred at a normal rate, 80% of them did not become gravid. In the few gravid infected mice, implantation numbers were as in uninfected control mice, but 28% of fetuses resorbed. Such infertility and early fetal losses we...

  13. Complex I (NADH:ubiquinone oxidoreductase) is active in but non-essential for procyclic Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Verner, Zdeněk; Čermáková, P.; Škodová, Ingrid; Kriegová, Eva; Horváth, A.; Lukeš, Julius

    2011-01-01

    Roč. 175, č. 2 (2011), s. 196-200 ISSN 0166-6851 R&D Projects: GA ČR GA204/09/1667; GA ČR GD206/09/H026; GA MŠk 2B06129; GA MŠk LC07032 Institutional research plan: CEZ:AV0Z60220518 Keywords : Trypanosoma * Mitochondrion * Respiration * Complex I Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.551, year: 2011

  14. Alternative NADH dehydrogenase (NDH2): intermembrane-space-facing counterpart of mitochondrial complex I in the procyclic Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Verner, Zdeněk; Škodová, Ingrid; Poláková, S.; Ďurišová-Benkovičková, V.; Horváth, A.; Lukeš, Julius

    2013-01-01

    Roč. 140, č. 3 (2013), s. 328-337 ISSN 0031-1820 R&D Projects: GA MŠk LC07032; GA ČR GA204/09/1667 Institutional support: RVO:60077344 Keywords : Trypanosoma * mitochondrion * dehydrogenase * respiration * NDH2 Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.350, year: 2013 http://journals.cambridge.org/action/displayAbstract?fromPage=online&aid=8838254

  15. The assembly of F1FO-ATP synthase is disrupted upon interference of RNA editing in Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Hashimi, Hassan; Benkovičová, V.; Čermáková, P.; Lai, De Hua; Horváth, A.; Lukeš, Julius

    2010-01-01

    Roč. 40, č. 1 (2010), s. 45-54 ISSN 0020-7519 R&D Projects: GA ČR GA204/06/1558; GA AV ČR IAA500960705 Institutional research plan: CEZ:AV0Z60220518 Keywords : RNA editing * ATP synthase * mitochondrion * Trypanosoma * respiratory complex * membrane potential Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.822, year: 2010

  16. Dynamics of Mitochondrial RNA-Binding Protein Complex in Trypanosoma brucei and Its Petite Mutant under Optimized Immobilization Conditions

    Czech Academy of Sciences Publication Activity Database

    Huang, Zhenqiu; Kaltenbrunner, S.; Šimková, Eva; Staněk, David; Lukeš, Julius; Hashimi, Hassan

    2014-01-01

    Roč. 13, č. 9 (2014), s. 1232-1240 ISSN 1535-9778 R&D Projects: GA ČR GAP305/12/2261; GA MŠk(CZ) EE2.3.30.0032 Institutional support: RVO:60077344 ; RVO:68378050 Keywords : mitochondrion * Trypanosoma brucei * YFP Subject RIV: EB - Genetics ; Molecular Biology; EB - Genetics ; Molecular Biology (UMG-J) Impact factor: 2.820, year: 2014

  17. Mitochondrial localization of human frataxin is necessary but processing is not for rescuing frataxin deficiency in Trypanosoma brucei

    Czech Academy of Sciences Publication Activity Database

    Long, Shaojun; Jirků, Milan; Ayala, F. J.; Lukeš, Julius

    2008-01-01

    Roč. 105, č. 36 (2008), s. 13468-13473 ISSN 0027-8424 R&D Projects: GA AV ČR IAA500960705; GA MŠk LC07032; GA MŠk 2B06129; GA ČR GA204/06/1558 Institutional research plan: CEZ:AV0Z60220518 Keywords : frataxin * mitochondrion * Trypanosoma * Kinetoplastida Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 9.380, year: 2008

  18. Hosts and vectors of Trypanosoma cruzi discrete typing units in the Chagas disease endemic region of the Paraguayan Chaco

    OpenAIRE

    ACOSTA, NIDIA; L?PEZ, ELSA; LEWIS, MICHAEL D.; LLEWELLYN, MARTIN S.; G?MEZ, ANA; ROM?N, FABIOLA; MILES, MICHAEL A.; YEO, MATTHEW

    2017-01-01

    SUMMARY Active Trypanosoma cruzi transmission persists in the Gran Chaco region, which is considered hyperendemic for Chagas disease. Understanding domestic and sylvatic transmission cycles and therefore the relationship between vectors and mammalian hosts is crucial to designing and implementing improved effective control strategies. Here we describe the species of triatomine vectors and the sylvatic mammal reservoirs of T. cruzi, in different localities of the Paraguayan and Bolivian Chaco....

  19. Infecção via oral por Trypanosoma evansi em animais de laboratório Oral infection by Trypanosoma evansi in rats and mice

    Directory of Open Access Journals (Sweden)

    Aleksandro Schafer da Silva

    2007-06-01

    Full Text Available Testou-se a infecção de Trypanosoma evansi pela via oral em ratos e camundongos, através de sangue contaminado de ambas as espécies. Dez ratos e dez camundongos foram alocados em quatro grupos iguais A e B (ratos, C e D (camundongos. Os grupos A e C receberam sangue contaminado de um rato e o grupo B e D de um camundongo, através de uma sonda. O volume de sangue administrado foi de 0,2ml, o qual apresentava uma concentração de 10(7 tripanossomas ml-1. Os animais foram mantidos em temperatura e umidade constantes (25°C e 80% UR, sendo realizados esfregaços sanguíneos diários para identificar o período pré-patente e a evolução do parasita na circulação. Nos grupos A e B, o período pré-patente variou de 19 a 25 dias, e o período entre a detecção dos parasitas e a morte dos animais foi em média de 12,7 dias. Os camundongos do grupo C e D não apresentaram infecção pelo parasita, sendo estes avaliados por 60 dias. Os ratos foram susceptíveis a infecção por T. evansi pela via oral; entretanto, os camundongos não se contaminaram com o protozoário por via digestiva.In this research, Trypanosoma evansi infection was tested in rats and mice by oral ingestion of contaminated blood. Groups of ten rats and ten mice were disposed in four experimental groups: A and B (rats, C and D (mice. The groups A and C were contaminated by rat-contaminated blood; B and C groups by mouse-contaminated blood. The blood was given using a probe filled with 0.2ml of contaminated blood with 10(7 trypanosomes ml-1. These animals were maintained at constant temperature and humidity (25°C and 80% UR. Dairy blood smear were done to identify the prepatent period and evolution of parasite in the circulation. In the A and B groups, the pre latency period varied from 19 to 25 days and the period of parasite detection and animals death was an average of 12.7 days. The C and D groups did not present infection by the parasite even when evaluated for 60 days

  20. Risk Factors Associated with Triatomines and Its Infection with Trypanosoma cruzi in Rural Communities from the Southern Region of the State of Mexico, Mexico

    Science.gov (United States)

    Medina-Torres, Imelda; Vázquez-Chagoyán, Juan C.; Rodríguez-Vivas, Roger I.; de Oca-Jiménez, Roberto Montes

    2010-01-01

    Trypanosoma cruzi prevalence in triatomines and risk factors associated to the presence of the insect were studied in 990 rural houses in the southern region of the State of Mexico, Mexico. In each house, triatomines were collected, and information related to house construction material was obtained. T. cruzi infection was diagnosed in all triatomines. A primary screening was performed using 2 × 2 contingency tables of exposure variables. All variables with P ≤ 0.20 were analyzed by logistic regression. Triatomines (N = 125) were collected from 822 houses and analyzed for T. cruzi infection. Triatoma pallidipennis (97.4%) and Triatoma dimidiata (2.6%) were identified in 52.1% of the localities and in 6.1% of the houses. Infection was found in 28.0% of triatomines, from which 28.9% were nymphs. Factors associated with triatomine infestation were flooring construction material (dirt floor: odds ratio [OR], 10.05; 95% confidence interval [CI], 5.31–18.04; P = 0.0001), house rooms (at least three rooms: OR, 2.04; 95% CI, 1.07–3.86; P = 0.028), and ceiling construction material (cardboard lamina tile: OR, 6.84; 95% CI, 1.49–31.31; P = 0.013). This study shows T. cruzi circulation in triatomines in the area of study, and because triatomines are adapted for living and reproducing in the domestic environment, there is a potential risk of Chagas disease transmission to humans. Also, we can conclude that the construction materials and house inhabitants are risk factors of triatomines infestation. PMID:20064995

  1. Prevalence of Chagas disease in pregnant women and congenital transmission of Trypanosoma cruzi in Brazil: a systematic review and meta-analysis.

    Science.gov (United States)

    Martins-Melo, Francisco Rogerlândio; Lima, Mauricélia da Silveira; Ramos, Alberto Novaes; Alencar, Carlos Henrique; Heukelbach, Jörg

    2014-08-01

    To estimate the prevalence of Chagas disease in pregnant women and the risk of congenital transmission of Trypanosoma cruzi infection in Brazil, through a systematic review and meta-analysis. We searched electronic databases, grey literature and reference lists of included publications to identify epidemiological studies on the prevalence of Chagas disease in pregnant women and on the congenital transmission rate of T. cruzi infection in Brazil published between January 1980 and June 2013. Pooled estimates and 95% confidence intervals (95% CIs) were calculated using fixed- and random-effects models. Sixteen articles were included - 12 studies on the prevalence of Chagas disease in pregnant women (549,359 pregnant women) and nine on congenital transmission rates (1687 children born to infected mothers). Prevalence of Chagas disease in pregnant women ranged from 0.1% to 8.5%, and congenital transmission rates from 0% to 5.2%. The pooled prevalence of Chagas disease among pregnant women across studies was 1.1% (95% CI: 0.6-2.0); the pooled congenital transmission rate was 1.7% (95% CI: 0.9-3.1). In 2010, 34,629 pregnant women were estimated to be infected with T. cruzi, and 312-1073 children born (mean: 589 cases) with congenital infection. Congenital Chagas disease is a neglected public health problem in Brazil. Systematic congenital Chagas disease control programs through routine prenatal screening for T. cruzi should be widely implemented in Brazil's endemic areas, to identify infected pregnant women and newborns at risk of congenital infection. © 2014 John Wiley & Sons Ltd.

  2. Prevalence of antibodies against Trypanosoma cruzi in pregnant women in endemic areas of the department of Boyacá, Colombia

    Directory of Open Access Journals (Sweden)

    Suescún-Carrero, Sandra Helena

    2017-10-01

    Full Text Available Objective: To determine the prevalence of antibodies against Trypanosoma cruzi in pregnant women in endemic areas of Boyacá, Colombia, in 2012 and 2013. Materials and methods: Cross-sectional study of 566 pregnant women from endemic municipalities of Boyacá. Samples were analyzed by means of serological tests for Chagas, namely: IgG ELISA, indirect immunofluorescence and indirect hemagglutination. Cases with positive results in two tests were considered as confirmed. Results: The overall prevalence of antibodies against Trypanosoma cruzi was 2.5 % (14/566. Municipalities with the highest prevalence were Chitaraque (8.3 %, and Soatá (3.3 %. Average age of positive women was 32.6 years, and their gestational period, 18.1 weeks. We found a statistically significant association between age and the presence of antibodies against Trypanosoma cruzi. Conclusion: Prevalence of antibodies against T. cruzi in pregnant women demonstrates the importance of the monitoring program for Chagas disease in pregnancy, as a method for congenital disease control.

  3. Fission yield measurements at IGISOL

    Science.gov (United States)

    Lantz, M.; Al-Adili, A.; Gorelov, D.; Jokinen, A.; Kolhinen, V. S.; Mattera, A.; Moore, I.; Penttilä, H.; Pomp, S.; Prokofiev, A. V.; Rakopoulos, V.; Rinta-Antila, S.; Simutkin, V.; Solders, A.

    2016-06-01

    The fission product yields are an important characteristic of the fission process. In fundamental physics, knowledge of the yield distributions is needed to better understand the fission process. For nuclear energy applications good knowledge of neutroninduced fission-product yields is important for the safe and efficient operation of nuclear power plants. With the Ion Guide Isotope Separator On-Line (IGISOL) technique, products of nuclear reactions are stopped in a buffer gas and then extracted and separated by mass. Thanks to the high resolving power of the JYFLTRAP Penning trap, at University of Jyväskylä, fission products can be isobarically separated, making it possible to measure relative independent fission yields. In some cases it is even possible to resolve isomeric states from the ground state, permitting measurements of isomeric yield ratios. So far the reactions U(p,f) and Th(p,f) have been studied using the IGISOL-JYFLTRAP facility. Recently, a neutron converter target has been developed utilizing the Be(p,xn) reaction. We here present the IGISOL-technique for fission yield measurements and some of the results from the measurements on proton induced fission. We also present the development of the neutron converter target, the characterization of the neutron field and the first tests with neutron-induced fission.

  4. Fission yield measurements at IGISOL

    Directory of Open Access Journals (Sweden)

    Lantz M.

    2016-01-01

    Full Text Available The fission product yields are an important characteristic of the fission process. In fundamental physics, knowledge of the yield distributions is needed to better understand the fission process. For nuclear energy applications good knowledge of neutroninduced fission-product yields is important for the safe and efficient operation of nuclear power plants. With the Ion Guide Isotope Separator On-Line (IGISOL technique, products of nuclear reactions are stopped in a buffer gas and then extracted and separated by mass. Thanks to the high resolving power of the JYFLTRAP Penning trap, at University of Jyväskylä, fission products can be isobarically separated, making it possible to measure relative independent fission yields. In some cases it is even possible to resolve isomeric states from the ground state, permitting measurements of isomeric yield ratios. So far the reactions U(p,f and Th(p,f have been studied using the IGISOL-JYFLTRAP facility. Recently, a neutron converter target has been developed utilizing the Be(p,xn reaction. We here present the IGISOL-technique for fission yield measurements and some of the results from the measurements on proton induced fission. We also present the development of the neutron converter target, the characterization of the neutron field and the first tests with neutron-induced fission.

  5. Trypanosoma evansi

    African Journals Online (AJOL)

    Parasitaemia in the untreated control and the treated rats were monitored using Haematocrit Centrifuge Technique. (HCT) twice weekly. Two rats from each group were sacrificed at 45 dpi; visceral ... Protozoology laboratory of the Department of. Veterinary Parasitology and Entomology, Ahmadu Bello. University, Zaria for ...

  6. Contribución al conocimiento de los reservorios del Trypanosoma cruzi (Chagas,1909 en la Provincia de Corrientes, Argentina Contribution to knowledge of reservoirs of Trypanosoma cruzi (Chagas, 1909 in Corrientes Province, Argentina

    Directory of Open Access Journals (Sweden)

    María Esther Bar

    1999-06-01

    Full Text Available Con el propósito de identificar a reservorios del Trypanosoma cruzi se investigaron 60 mamíferos en los Departamentos Capital y San Luis del Palmar. Se examinaron: primates, roedores, marsupiales, carnívoros y edentados; 40 vivían en cautiverio y 20 fueron capturados mediante trampas en una comunidad rural forestal. Los mamíferos fueron analizados por xenodiagnóstico, empleándose ninfas de 3o o 4o estadío de Triatoma infestans ayunadas durante 2 semanas. Las heces de los triatominos fueron observadas al microscopio (400x a los 30, 60 y 90 días post-alimentación. En 2 Saimiri sciureus y en 1 Cebus apella se constató infección por tripanosomas cruziformes. Se concluye que la parasitemia detectada fue baja. La presencia de Didelphis albiventris, reservorio potencial del Trypanosoma cruzi , en una zona de transmisión activa del parásito representa un factor de riesgo, por lo que son necesarias futuras investigaciones epidemiológicas para determinar la real diagnosis de esta parasitosis en la provincia de Corrientes, Argentina.In order to identify Trypanosoma cruzi reservoirs in transmission areas, 60 mammals in Capital and San Luis del Palmar Departments, Corrientes, Argentina were studied. Primates, rodents, carnivores, marsupials and edentates were investigated, 40 of them living in captivity and 20 caught with traps in a rural area. The mammals were examined by xenodiagnosis and third or fourth instars nymphs of Triatoma infestans starved for 2 weeks were used. The feces were microscopically observed (400x for Trypanosoma cruzi infection at 30, 60 and 90 days after feeding. Trypanosoma cruzi-like parasites were identified in 2 Saimiri sciureus and 1 Cebus apella analyzed by xenodiagnosis. It was concluded that parasitemia was low. Howewer, the presence in a forest area of Didelphis albiventris, potential reservoir of the parasite, indicates a risk factor and deserves further epidemiological study for a true diagnosis of this

  7. Cherry tomato yield in greenhouses with different plastic covers

    Directory of Open Access Journals (Sweden)

    Ester Holcman

    2017-08-01

    Full Text Available ABSTRACT: The objective of the present study was to evaluate the influence of different plastic covers on microclimate and cherry tomato yield in greenhouses. The experiments were carried out in Piracicaba, state of São Paulo (Brazil, during three growing periods (2008/2009/2010. A greenhouse was divided in: Environment I (EI - covered with plastic film anti-UV and thermo-reflective shading screen, and Environment II (EII - covered with diffusive plastic film; monitored with automatic weather sensors; and cultivated with cherry tomato (‘Sweet Grape’ and ‘Sweet Million’. Use of diffusive plastic in greenhouses provides a better inside distribution of solar energy without causing major changes in air temperature and relative humidity, resulting in higher yield (kg plant-1, fruits quantity (number plant-1 and fruits average weight than those obtained under thermo-reflective shading screen.

  8. Reseña Histórica de algunos estudios Colombianos sobre Trypanosoma rangeli

    Directory of Open Access Journals (Sweden)

    Hernando Groot Liévano

    2000-08-01

    Full Text Available

    Parecerá extraño que uno de los primeros artículos de esta revista no se refiera a la enfermedad de Chagas. Tanto es así que cuando el doctor Felipe Guhl tuvo la idea de llamarme para esta presentación, mi primera respuesta fue negativa porque, obviamente debería hablarse del Trypanosoma cruzi y no del Trypanosoma rangeli. El presente artículo es un breve recuento de mi experiencia con este parásito y su importancia dado que coexiste con el T. cruzi y que, en ocasiones la diferenciación morfológica entre los dos no es tan clara cuando se examinan preparaciones de sangre en “gota gruesa” de vertebrados o preparaciones del contenido intestinal de los insectos vectores, y además porque tiene ciertas relaciones inmunológicas que es necesario tener en cuenta para evitar posibles confusiones. Por otra parte, su distribución geográfica es muy amplia extendiéndose desde México hasta el Perú y el Brasil.

    El Trypanosoma rangeli, llamado así por un distinguido médico y posteriormente diplomático de Venezuela, el doctor Enrique Tejera, quien encontró en los chipos, o sea en los Rhodnius prolixus de Venezuela, un pequeño flagelado muy largo, bastante diferente del Trypanosoma cruzi y resolvió ponerle el nombre de Trypanosoma o Crithidia rangeli pues no estaba muy seguro del género en el cual debía colocarlo. Evidentemente, sólo había visto la morfología de estos flagelados en el intestino de los Rhodnius y por consiguiente no tenía ningún otro elemento para identi-ficarlos.

    Únicamente comprobó que eran diferentes del cruzi. ¿Por qué le dedicó su descubrimiento a Rangel? Creo que es importante que nosotros los latinoamericanos conozcamos bien los valores científicos que han habido en nuestros países y en vez de preocuparnos por las artificiales fronteras políticas, lo cual en nada contribuye al progreso de la ciencia, comencemos a tener claro conocimiento de lo que en todas estas naciones hermanas por

  9. Experiências sôbre a transmissão do Trypanosoma cruzi por sanguessugas e de tripanosomas de vertebrados de sangue frio por triatomíneos Experiments of the transmission of Trypanosoma cruzi by leechs and cold blooded vertebrate trypanosomas by triatominae

    Directory of Open Access Journals (Sweden)

    Samuel B. Pessôa

    1969-06-01

    Full Text Available Observou-se que o Trypanosoma cruzi não se multiplica na sanguessuga (Haementeria lutzi Pinto; os tripanosomas sugados degeneram após algum tempo; outros permanecem aparentemente normais, porém 48 horas após a ingestão infectante acabam morrendo. Observou-se ainda que os tripanosomas parasitas da rã (T. rotatorium e T. leptodactyli bem como o T. hogei, parasita da serpente Rachidelus brazili, não se multiplicam no intestino dos triatomíneos. O mais resistente (o T. leptodactyli, permanece vivo até 72 horas após a ingestão infectante, porém as outras duas espécies (T. rotatorium e T. hogei não resistem mais de 24 horas após serem sugadas pelos triatomíneos.Trypanosoma cruzi does not reproduce itself in the leech (Haementerm lutzi Pinto; the ingested trypanosomes degenerate after some time; other organisms remain apparently normal, however dying 48 hours after the feeding of the leechs. The parasite trypanosomas of the frog (T. rotatorium and T. leptodactyli as well as those parasiting the ophidian Rachidelus brazili (T. hogei do not multiply in the intestine of the triatominae. The most resistent species (T. leptocbactyli remains alive 72 hours after the feeding of the triatominae; the other two, however, do not survive more than 24 hours.

  10. Increased genetic diversity and prevalence of co-infection with Trypanosoma spp. in koalas (Phascolarctos cinereus and their ticks identified using next-generation sequencing (NGS.

    Directory of Open Access Journals (Sweden)

    Amanda D Barbosa

    Full Text Available Infections with Trypanosoma spp. have been associated with poor health and decreased survival of koalas (Phascolarctos cinereus, particularly in the presence of concurrent pathogens such as Chlamydia and koala retrovirus. The present study describes the application of a next-generation sequencing (NGS-based assay to characterise the prevalence and genetic diversity of trypanosome communities in koalas and two native species of ticks (Ixodes holocyclus and I. tasmani removed from koala hosts. Among 168 koalas tested, 32.2% (95% CI: 25.2-39.8% were positive for at least one Trypanosoma sp. Previously described Trypanosoma spp. from koalas were identified, including T. irwini (32.1%, 95% CI: 25.2-39.8%, T. gilletti (25%, 95% CI: 18.7-32.3%, T. copemani (27.4%, 95% CI: 20.8-34.8% and T. vegrandis (10.1%, 95% CI: 6.0-15.7%. Trypanosoma noyesi was detected for the first time in koalas, although at a low prevalence (0.6% 95% CI: 0-3.3%, and a novel species (Trypanosoma sp. AB-2017 was identified at a prevalence of 4.8% (95% CI: 2.1-9.2%. Mixed infections with up to five species were present in 27.4% (95% CI: 21-35% of the koalas, which was significantly higher than the prevalence of single infections 4.8% (95% CI: 2-9%. Overall, a considerably higher proportion (79.7% of the Trypanosoma sequences isolated from koala blood samples were identified as T. irwini, suggesting this is the dominant species. Co-infections involving T. gilletti, T. irwini, T. copemani, T. vegrandis and Trypanosoma sp. AB-2017 were also detected in ticks, with T. gilletti and T. copemani being the dominant species within the invertebrate hosts. Direct Sanger sequencing of Trypanosoma 18S rRNA gene amplicons was also performed and results revealed that this method was only able to identify the genotypes with greater amount of reads (according to NGS within koala samples, which highlights the advantages of NGS in detecting mixed infections. The present study provides new insights

  11. Increased genetic diversity and prevalence of co-infection with Trypanosoma spp. in koalas (Phascolarctos cinereus) and their ticks identified using next-generation sequencing (NGS).

    Science.gov (United States)

    Barbosa, Amanda D; Gofton, Alexander W; Paparini, Andrea; Codello, Annachiara; Greay, Telleasha; Gillett, Amber; Warren, Kristin; Irwin, Peter; Ryan, Una

    2017-01-01

    Infections with Trypanosoma spp. have been associated with poor health and decreased survival of koalas (Phascolarctos cinereus), particularly in the presence of concurrent pathogens such as Chlamydia and koala retrovirus. The present study describes the application of a next-generation sequencing (NGS)-based assay to characterise the prevalence and genetic diversity of trypanosome communities in koalas and two native species of ticks (Ixodes holocyclus and I. tasmani) removed from koala hosts. Among 168 koalas tested, 32.2% (95% CI: 25.2-39.8%) were positive for at least one Trypanosoma sp. Previously described Trypanosoma spp. from koalas were identified, including T. irwini (32.1%, 95% CI: 25.2-39.8%), T. gilletti (25%, 95% CI: 18.7-32.3%), T. copemani (27.4%, 95% CI: 20.8-34.8%) and T. vegrandis (10.1%, 95% CI: 6.0-15.7%). Trypanosoma noyesi was detected for the first time in koalas, although at a low prevalence (0.6% 95% CI: 0-3.3%), and a novel species (Trypanosoma sp. AB-2017) was identified at a prevalence of 4.8% (95% CI: 2.1-9.2%). Mixed infections with up to five species were present in 27.4% (95% CI: 21-35%) of the koalas, which was significantly higher than the prevalence of single infections 4.8% (95% CI: 2-9%). Overall, a considerably higher proportion (79.7%) of the Trypanosoma sequences isolated from koala blood samples were identified as T. irwini, suggesting this is the dominant species. Co-infections involving T. gilletti, T. irwini, T. copemani, T. vegrandis and Trypanosoma sp. AB-2017 were also detected in ticks, with T. gilletti and T. copemani being the dominant species within the invertebrate hosts. Direct Sanger sequencing of Trypanosoma 18S rRNA gene amplicons was also performed and results revealed that this method was only able to identify the genotypes with greater amount of reads (according to NGS) within koala samples, which highlights the advantages of NGS in detecting mixed infections. The present study provides new insights on the

  12. RNA-Seq analysis validates the use of culture-derived Trypanosoma brucei and provides new markers for mammalian and insect life-cycle stages.

    Science.gov (United States)

    Naguleswaran, Arunasalam; Doiron, Nicholas; Roditi, Isabel

    2018-04-02

    Trypanosoma brucei brucei, the parasite causing Nagana in domestic animals, is closely related to the parasites causing sleeping sickness, but does not infect humans. In addition to its importance as a pathogen, the relative ease of genetic manipulation and an innate capacity for RNAi extend its use as a model organism in cell and infection biology. During its development in its mammalian and insect (tsetse fly) hosts, T. b. brucei passes through several different life-cycle stages. There are currently four life-cycle stages that can be cultured: slender forms and stumpy forms, which are equivalent to forms found in the mammal, and early and late procyclic forms, which are equivalent to forms in the tsetse midgut. Early procyclic forms show coordinated group movement (social motility) on semi-solid surfaces, whereas late procyclic forms do not. RNA-Seq was performed on biological replicates of each life-cycle stage. These constitute the first datasets for culture-derived slender and stumpy bloodstream forms and early and late procyclic forms. Expression profiles confirmed that genes known to be stage-regulated in the animal and insect hosts were also regulated in culture. Sequence reads of 100-125 bases provided sufficient precision to uncover differential expression of closely related genes. More than 100 transcripts showed peak expression in stumpy forms, including adenylate cyclases and several components of inositol metabolism. Early and late procyclic forms showed differential expression of 73 transcripts, a number of which encoded proteins that were previously shown to be stage-regulated. Moreover, two adenylate cyclases previously shown to reduce social motility are up-regulated in late procyclic forms. This study validates the use of cultured bloodstream forms as alternatives to animal-derived parasites and yields new markers for all four stages. In addition to underpinning recent findings that early and late procyclic forms are distinct life-cycle stages

  13. Evaluation of Yield and Yield Attributes of Five Sweet Potato ...

    African Journals Online (AJOL)

    0087, and TIS 2532.OP.1.13) were evaluated for yield and agronomic performance in Imo State University Farm, Owerri. The experiment was laid out in a randomised complete block design with three replications. The planting density was 33,000 ...

  14. Heterosis and combining ability for grain yield and yield component ...

    African Journals Online (AJOL)

    ... ranged from 0 to -13% indicating that the hybrids tend to be earlier in maturity than the parents. The mean squares due to GCA for days to maturity, ear diameter, member of kernels per row, 1000 kernel weight and grain yield were significant, indicating the importance of additive genetic variance in controlling these traits.

  15. Rice yield prediction from yield components and limiting factors

    NARCIS (Netherlands)

    Casanova, D.; Goudriaan, J.; Catala Former, M.M.; Withagen, J.C.M.

    2002-01-01

    This article aims to quantify growth at field level in relation to crop status and soil properties in irrigated direct-seeded rice. Forty fields were selected in the Ebro Delta (Spain). Rice growth was monitored and soil properties measured. Yield was related to soil properties by a deductive

  16. Correlation Analysis of some Growth, Yield, Yield Components and ...

    African Journals Online (AJOL)

    three critical growth stages which was imposed by withholding water (at ... November, 5th December, 19th December and 2nd January) laid out in a split ... Simple correlation coefficient ® of different crop parameters and grain yield ... The husk bran and germ are rich sources of ..... heat in 2009/2010 dry season at Fadam a ...

  17. Diagnostic yield of molecular autopsy in patients with sudden arrhythmic death syndrome using targeted exome sequencing

    DEFF Research Database (Denmark)

    Nunn, Laurence M; Lopes, Luis R; Syrris, Petros

    2016-01-01

    AIMS: The targeted genetic screening of Sudden Arrhythmic Death Syndrome (SADS) probands in a molecular autopsy has a diagnostic yield of up to 35%. Exome sequencing has the potential to improve this yield. The primary aim of this study is to examine the feasibility and diagnostic utility...... of targeted exome screening in SADS victims, utilizing familial clinical screening whenever possible. METHODS AND RESULTS: To determine the feasibility and diagnostic yield of targeted exome sequencing deoxyribonucleic acid (DNA) was isolated from 59 SADS victims (mean age 25 years, range 1-51 years...... previously published rare (0.02-0.5%) candidate mutations-a total yield of 29%. Co-segregation fully confirmed two private SCN5A Na channel mutations. Variants of unknown significance were detected in a further 34% of probands. CONCLUSION: Molecular autopsy using targeted exome sequencing has a relatively...

  18. Status of fission yield data

    International Nuclear Information System (INIS)

    England, T.R.; Blachot, J.

    1988-01-01

    In this paper we summarize the current status of the recent US evaluation for 34 fissioning nuclides at one or more neutron incident energies and for spontaneous fission. Currently there are 50 yields sets, and for each we have independent and cumulative yields and uncertainties for approximately 1100 fission products. When finalized the recommended data will become part of Version VI of the US ENDF/B. Other major evaluations in progress that are included in a recently formed IAEA Coordinated Research Program are also summarized. In a second part we review two empirical models in use to estimate independent yields. Comparison of model estimates with measured data is presented, including a comparison with some recent data obtained from Lohengrin (Cf-249 T). 18 refs., 13 figs., 3 tabs

  19. Trypanosoma cruzi in dogs: electrocardiographic and echocardiographic evaluation, in Malinalco, State of Mexico

    Directory of Open Access Journals (Sweden)

    González-Vieyra SD

    2011-12-01

    Full Text Available Sandra Díaz González-Vieyra1, Ninfa Ramírez-Durán2, Ángel H Sandoval-Trujillo3, Juan C Vázquez-Chagoyán1, Humberto G Monroy-Salazar1, Alberto Barbabosa-Pliego11Research Center of Advanced Studies in Animal Health, Veterinary Husbandry School, 2Medical and Ambiental Microbiology, Research Center of Advanced Studies in Health Science, School of Medicine, Autonomous University of the State of Mexico, Toluca, Mexico; 3Department of Biological Systems, Metropolitan Autonomous University, Xochimilco, Mexico City, MexicoAbstract: Chagas disease caused by Trypanosoma cruzi is an important public health problem in Latin America. Dogs are considered a risk factor for human Chagas disease, a sentinel for T. cruzi infection in endemic regions and an animal model to study pathological aspects of the disease. The potential use of dogs as indicators of human cardiac pathogenicity of local T. cruzi strains has been studied insufficiently. We studied electrocardiographic (EKG and echocardiographic (ECG alteration frequencies observed in an open population of dogs in Malinalco, Mexico, and determined if such frequencies were statistically associated with T. cruzi infection in dogs. Animals (n = 139 were clinically examined and owners were asked to answer a questionnaire about dogs’ living conditions. Two commercial serological tests (IHA, ELISA were conducted to detect anti-T. cruzi serum antibodies. Significant differences between seropositive and seronegative animals in cardiomyopathic frequencies were detected through EKG and ECG (P < 0.05. Thirty dogs (21.58% were serologically positive to anti-T. cruzi antibodies (to ELISA and IHA assays, of which nine (30% had EKG and/or ECG alterations. From the remaining 104 (78.42% seronegative animals, five (4.5% had EKG and/or ECG abnormalities. Our data support the hypothesis that most EKG and ECG alterations found in dogs from Malinalco could be associated with T. cruzi infection. Considering the dog as a

  20. Distantiae transmission of Trypanosoma cruzi: a new epidemiological feature of acute Chagas disease in Brazil.

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    Samanta Cristina das Chagas Xavier

    2014-05-01

    Full Text Available BACKGROUND: The new epidemiological scenario of orally transmitted Chagas disease that has emerged in Brazil, and mainly in the Amazon region, needs to be addressed with a new and systematic focus. Belém, the capital of Pará state, reports the highest number of acute Chagas disease (ACD cases associated with the consumption of açaí juice. METHODOLOGY/PRINCIPAL FINDINGS: The wild and domestic enzootic transmission cycles of Trypanosoma cruzi were evaluated in the two locations (Jurunas and Val-de Cães that report the majority of the autochthonous cases of ACD in Belém city. Moreover, we evaluated the enzootic cycle on the three islands that provide most of the açaí fruit that is consumed in these localities. We employed parasitological and serological tests throughout to evaluate infectivity competence and exposure to T. cruzi. In Val-de-Cães, no wild mammal presented positive parasitological tests, and 56% seroprevalence was observed, with low serological titers. Three of 14 triatomines were found to be infected (TcI. This unexpected epidemiological picture does not explain the high number of autochthonous ACD cases. In Jurunas, the cases of ACD could not be autochthonous because of the absence of any enzootic cycle of T. cruzi. In contrast, in the 3 island areas from which the açaí fruit originates, 66.7% of wild mammals and two dogs displayed positive hemocultures, and 15.6% of triatomines were found to be infected by T. cruzi. Genotyping by mini-exon gene and PCR-RFLP (1f8/Akw21I targeting revealed that the mammals and triatomines from the islands harbored TcI and Trypanosoma rangeli in single and mixed infections. CONCLUSION/SIGNIFICANCE: These findings show that cases of Chagas disease in the urban area of Belém may be derived from infected triatomines coming together with the açaí fruits from distant islands. We term this new epidemiological feature of Chagas disease as "Distantiae transmission".

  1. Trypanocide Treatment of Women Infected with Trypanosoma cruzi and Its Effect on Preventing Congenital Chagas

    Science.gov (United States)

    Fabbro, Diana L.; Danesi, Emmaria; Olivera, Veronica; Codebó, Maria Olenka; Denner, Susana; Heredia, Cecilia; Streiger, Mirtha; Sosa-Estani, Sergio

    2014-01-01

    With the control of the vectorial and transfusional routes of infection with Trypanosoma cruzi, congenital transmission has become an important source of new cases. This study evaluated the efficacy of trypanocidal therapy to prevent congenital Chagas disease and compared the clinical and serological evolution between treated and untreated infected mothers. We conducted a multicenter, observational study on a cohort of mothers infected with T. cruzi, with and without trypanocidal treatment before pregnancy. Their children were studied to detect congenital infection. Among 354 “chronically infected mother-biological child” pairs, 132 were treated women and 222 were untreated women. Among the children born to untreated women, we detected 34 infected with T. cruzi (15.3%), whose only antecedent was maternal infection. Among the 132 children of previously treated women, no infection with T. cruzi was found (0.0%) (p<0.05). Among 117 mothers with clinical and serological follow up, 71 had been treated and 46 were untreated. The women were grouped into three groups. Group A: 25 treated before 15 years of age; Group B: 46 treated at 15 or more years of age; Group C: untreated, average age of 29.2±6.2 years at study entry. Follow-up for Groups A, B and C was 16.3±5.8, 17.5±9.2 and 18.6±8.6 years respectively. Negative seroconversion: Group A, 64.0% (16/25); Group B, 32.6% (15/46); Group C, no seronegativity was observed. Clinical electrocardiographic alterations compatible with chagasic cardiomyopathy: Group A 0.0% (0/25); B 2.2% (1/46) and C 15.2% (7/46). The trypanocidal treatment of women with chronic Chagas infection was effective in preventing the congenital transmission of Trypanosoma cruzi to their children; it had also a protective effect on the women's clinical evolution and deparasitation could be demonstrated in many treated women after over 10 years of follow up. PMID:25411847

  2. Channel-forming activities in the glycosomal fraction from the bloodstream form of Trypanosoma brucei.

    Directory of Open Access Journals (Sweden)

    Melisa Gualdron-López

    Full Text Available BACKGROUND: Glycosomes are a specialized form of peroxisomes (microbodies present in unicellular eukaryotes that belong to the Kinetoplastea order, such as Trypanosoma and Leishmania species, parasitic protists causing severe diseases of livestock and humans in subtropical and tropical countries. The organelles harbour most enzymes of the glycolytic pathway that is responsible for substrate-level ATP production in the cell. Glycolysis is essential for bloodstream-form Trypanosoma brucei and enzymes comprising this pathway have been validated as drug targets. Glycosomes are surrounded by a single membrane. How glycolytic metabolites are transported across the glycosomal membrane is unclear. METHODS/PRINCIPAL FINDINGS: We hypothesized that glycosomal membrane, similarly to membranes of yeast and mammalian peroxisomes, contains channel-forming proteins involved in the selective transfer of metabolites. To verify this prediction, we isolated a glycosomal fraction from bloodstream-form T. brucei and reconstituted solubilized membrane proteins into planar lipid bilayers. The electrophysiological characteristics of the channels were studied using multiple channel recording and single channel analysis. Three main channel-forming activities were detected with current amplitudes 70-80 pA, 20-25 pA, and 8-11 pA, respectively (holding potential +10 mV and 3.0 M KCl as an electrolyte. All channels were in fully open state in a range of voltages ±150 mV and showed no sub-conductance transitions. The channel with current amplitude 20-25 pA is anion-selective (P(K+/P(Cl-∼0.31, while the other two types of channels are slightly selective for cations (P(K+/P(Cl- ratios ∼1.15 and ∼1.27 for the high- and low-conductance channels, respectively. The anion-selective channel showed an intrinsic current rectification that may suggest a functional asymmetry of the channel's pore. CONCLUSIONS/SIGNIFICANCE: These results indicate that the membrane of glycosomes

  3. Trypanocide treatment of women infected with Trypanosoma cruzi and its effect on preventing congenital Chagas.

    Science.gov (United States)

    Fabbro, Diana L; Danesi, Emmaria; Olivera, Veronica; Codebó, Maria Olenka; Denner, Susana; Heredia, Cecilia; Streiger, Mirtha; Sosa-Estani, Sergio

    2014-11-01

    With the control of the vectorial and transfusional routes of infection with Trypanosoma cruzi, congenital transmission has become an important source of new cases. This study evaluated the efficacy of trypanocidal therapy to prevent congenital Chagas disease and compared the clinical and serological evolution between treated and untreated infected mothers. We conducted a multicenter, observational study on a cohort of mothers infected with T. cruzi, with and without trypanocidal treatment before pregnancy. Their children were studied to detect congenital infection. Among 354 "chronically infected mother-biological child" pairs, 132 were treated women and 222 were untreated women. Among the children born to untreated women, we detected 34 infected with T. cruzi (15.3%), whose only antecedent was maternal infection. Among the 132 children of previously treated women, no infection with T. cruzi was found (0.0%) (p<0.05). Among 117 mothers with clinical and serological follow up, 71 had been treated and 46 were untreated. The women were grouped into three groups. Group A: 25 treated before 15 years of age; Group B: 46 treated at 15 or more years of age; Group C: untreated, average age of 29.2 ± 6.2 years at study entry. Follow-up for Groups A, B and C was 16.3 ± 5.8, 17.5 ± 9.2 and 18.6 ± 8.6 years respectively. Negative seroconversion: Group A, 64.0% (16/25); Group B, 32.6% (15/46); Group C, no seronegativity was observed. Clinical electrocardiographic alterations compatible with chagasic cardiomyopathy: Group A 0.0% (0/25); B 2.2% (1/46) and C 15.2% (7/46). The trypanocidal treatment of women with chronic Chagas infection was effective in preventing the congenital transmission of Trypanosoma cruzi to their children; it had also a protective effect on the women's clinical evolution and deparasitation could be demonstrated in many treated women after over 10 years of follow up.

  4. Diminazene aceturate (Berenil modulates the host cellular and inflammatory responses to Trypanosoma congolense infection.

    Directory of Open Access Journals (Sweden)

    Shiby Kuriakose

    Full Text Available BACKGROUND: Trypanosoma congolense are extracellular and intravascular blood parasites that cause debilitating acute or chronic disease in cattle and other domestic animals. Diminazene aceturate (Berenil has been widely used as a chemotherapeutic agent for trypanosomiasis in livestock since 1955. As in livestock, treatment of infected highly susceptible BALB/c mice with Berenil leads to rapid control of parasitemia and survival from an otherwise lethal infection. The molecular and biochemical mechanisms of action of Berenil are still not very well defined and its effect on the host immune system has remained relatively unstudied. Here, we investigated whether Berenil has, in addition to its trypanolytic effect, a modulatory effect on the host immune response to Trypanosoma congolense. METHODOLOGY/PRINCIPAL FINDINGS: BALB/c and C57BL/6 mice were infected intraperitoneally with T. congolense, treated with Berenil and the expression of CD25 and FoxP3 on splenic cells was assessed directly ex vivo. In addition, serum levels and spontaneous and LPS-induced production of pro-inflammatory cytokines by splenic and hepatic CD11b⁺ cells were determined by ELISA. Berenil treatment significantly reduced the percentages of CD25⁺ cells, a concomitant reduction in the percentage of regulatory (CD4⁺Foxp3⁺ T cells and a striking reduction in serum levels of disease exacerbating pro-inflammatory cytokines including IL-6, IL-12, TNF and IFN-γ. Furthermore, Berenil treatment significantly suppressed spontaneous and LPS-induced production of inflammatory cytokines by splenic and liver macrophages and significantly ameliorated LPS-induced septic shock and the associated cytokine storm. CONCLUSIONS/SIGNIFICANCE: Collectively, these results provide evidence that in addition to its direct trypanolytic effect, Berenil also modulates the host immune response to the parasite in a manner that dampen excessive immune activation and production of pathology

  5. Triatominae-Trypanosoma cruzi/T. rangeli: Vector-parasite interactions.

    Science.gov (United States)

    Vallejo, G A; Guhl, F; Schaub, G A

    2009-01-01

    Of the currently known 140 species in the family Reduviidae, subfamily Triatominae, those which are most important as vectors of the aetiologic agent of Chagas disease, Trypanosoma cruzi, belong to the tribes Triatomini and Rhodniini. The latter not only transmit T. cruzi but also Trypanosoma rangeli, which is considered apathogenic for the mammalian host but can be pathogenic for the vectors. Using different molecular methods, two main lineages of T. cruzi have been classified, T. cruzi I and T. cruzi II. Within T. cruzi II, five subdivisions are recognized, T. cruzi IIa-IIe, according to the variability of the ribosomal subunits 24Salpha rRNA and 18S rRNA. In T. rangeli, differences in the organization of the kinetoplast DNA separate two forms denoted T. rangeli KP1+ and KP1-, although differences in the intergenic mini-exon gene and of the small subunit rRNA (SSU rRNA) suggest four subpopulations denoted T. rangeli A, B, C and D. The interactions of these subpopulations of the trypanosomes with different species and populations of Triatominae determine the epidemiology of the human-infecting trypanosomes in Latin America. Often, specific subpopulations of the trypanosomes are transmitted by specific vectors in a particular geographic area. Studies centered on trypanosome-triatomine interaction may allow identification of co-evolutionary processes, which, in turn, could consolidate hypotheses of the evolution and the distribution of T. cruzi/T. rangeli-vectors in America, and they may help to identify the mechanisms that either facilitate or impede the transmission of the parasites in different vector species. Such mechanisms seem to involve intestinal bacteria, especially the symbionts which are needed by the triatomines to complete nymphal development and to produce eggs. Development of the symbionts is regulated by the vector. T. cruzi and T. rangeli interfere with this system and induce the production of antibacterial substances. Whereas T. cruzi is only

  6. Biochemical behavior of Trypanosoma cruzi strains isolated from mice submitted to specific chemotherapy

    Directory of Open Access Journals (Sweden)

    Jesila Pinto M. Marretto

    1994-12-01

    Full Text Available To investigate the influence of chemotherapy on the biochemical beha vior of Trypanosoma cruzi strains, three groups of mice were infected with one of three strains of T. cruzi of different biological and isoenzymic patterns (Peruvian, 21 SF and Colombian strains. Each group was subdivided into subgroups: 1 - treated with nifurtimox; 2 - treated with benznidazole and 3 - untreated infected controls. At the end of treatment, that lasted for 90 days, xenodiagnosis, sub inoculation of blood into new born mice and haemoculture were performed as tests of cure. From the positive tests, 22 samples of T. cruzi were isolated from all subgroups. Electrophoretic analysis of the isoenzymes PGM, GP1, ALAT and AS AT failed to show any difference between parasite strains isolated from treated and untreated mice, which indicates that no detectable clonal selection or parasite genetic markers alterations concerning the isoenzymes analysed have been determined by treatment with drugs of recognized antiparasitic effect, suggesting stability of the phenotypic characteristics of the three biological types of T. cruzi strains.Com o objetivo de investigar a influência da quimioterapia no padrão bioquímico de diferentes cepas do Trypanosoma cruzi, três grupos de camundongos foram infectados respectivamente com as cepas Peruana, 21 SF e Colombiana, que correspondem a diferentes padrões biológicos e isoenzimáticos. Cada grupo foi subdividido em subgrupos: 1 - tratados com nifurtimox; 2 - tratados com benzonidazol; 3- controles infectados não tratados. Ao final do tratamento que durou 90 dias, os animais foram submetidos a testes parasitológicos de cura: xenodiagnóstico, subinoculação do sangue em camundongos recém-nascidos e hemocultura em meio Warren. A partir da positivação destes testes, foram isoladas 22 amostras do T. cruzi dos três subgrupos. A análise eletroforética dos extratos enzimáticos obtidos após cultura para as enzimas PGM, GPI, ALAT e

  7. Trypanosoma cruzi Evades the Complement System as an Efficient Strategy to Survive in the Mammalian Host: The Specific Roles of Host/Parasite Molecules and Trypanosoma cruzi Calreticulin

    Directory of Open Access Journals (Sweden)

    Galia Ramírez-Toloza

    2017-09-01

    Full Text Available American Trypanosomiasis is an important neglected reemerging tropical parasitism, infecting about 8 million people worldwide. Its agent, Trypanosoma cruzi, exhibits multiple mechanisms to evade the host immune response and infect host cells. An important immune evasion strategy of T. cruzi infective stages is its capacity to inhibit the complement system activation on the parasite surface, avoiding opsonizing, immune stimulating and lytic effects. Epimastigotes, the non-infective form of the parasite, present in triatomine arthropod vectors, are highly susceptible to complement-mediated lysis while trypomastigotes, the infective form, present in host bloodstream, are resistant. Thus T. cruzi susceptibility to complement varies depending on the parasite stage (amastigote, trypomastigotes or epimastigote and on the T. cruzi strain. To avoid complement-mediated lysis, T. cruzi trypomastigotes express on the parasite surface a variety of complement regulatory proteins, such as glycoprotein 58/68 (gp58/68, T. cruzi complement regulatory protein (TcCRP, trypomastigote decay-accelerating factor (T-DAF, C2 receptor inhibitor trispanning (CRIT and T. cruzi calreticulin (TcCRT. Alternatively, or concomitantly, the parasite captures components with complement regulatory activity from the host bloodstream, such as factor H (FH and plasma membrane-derived vesicles (PMVs. All these proteins inhibit different steps of the classical (CP, alternative (AP or lectin pathways (LP. Thus, TcCRP inhibits the CP C3 convertase assembling, gp58/68 inhibits the AP C3 convertase, T-DAF interferes with the CP and AP convertases assembling, TcCRT inhibits the CP and LP, CRIT confers ability to resist the CP and LP, FH is used by trypomastigotes to inhibit the AP convertases and PMVs inhibit the CP and LP C3 convertases. Many of these proteins have similar molecular inhibitory mechanisms. Our laboratory has contributed to elucidate the role of TcCRT in the host

  8. Assessing potential sustainable wood yield

    Science.gov (United States)

    Robert F. Powers

    2001-01-01

    Society is making unprecedented demands on world forests to produce and sustain many values. Chief among them is wood supply, and concerns are rising globally about the ability of forests to meet increasing needs. Assessing this is not easy. It requires a basic understanding of the principles governing forest productivity: how wood yield varies with tree and stand...

  9. NIF total neutron yield diagnostic

    International Nuclear Information System (INIS)

    Cooper, Gary W.; Ruiz, Carlos L.

    2001-01-01

    We have designed a total neutron yield diagnostic for the National Ignition Facility (NIF) which is based on the activation of In and Cu samples. The particular approach that we have chosen is one in which we calibrate the entire counting system and which we call the ''F factor'' method. In this method, In and/or Cu samples are exposed to known sources of DD and DT neutrons. The activated samples are then counted with an appropriate system: a high purity Ge detector for In and a NaI coincidence system for Cu. We can then calculate a calibration factor, which relates measured activity to total neutron yield. The advantage of this approach is that specific knowledge of such quantities as cross sections and detector efficiencies is not needed. Unless the actual scattering environment of the NIF can be mocked up in the calibration experiment, the F factor will have to be modified using the results of a numerical simulation of the NIF scattering environment. In this article, the calibration factor methodology will be discussed and experimental results for the calibration factors will be presented. Total NIF neutron yields of 10 9 --10 19 can be measured with this method assuming a 50 cm stand-off distance can be employed for the lower yields

  10. Screening For Inhibitors Of Essential Leishmania Glucose Transporters

    Science.gov (United States)

    2011-07-01

    parasite life cycle and, unlike he amastigote form that lives inside mammalian macrophages, s viable provided that an alternative energy source such as pro...glucose transporters havebeenvalidated asnewdrug targets for proto- zoan parasites including Plasmodium falciparum, Leishmania mexicana and Trypanosoma...such as Leishmania species, Trypanosoma rucei, and Plasmodium falciparum, the causative agents of leish- aniasis, African sleeping sickness, and malaria

  11. Riesgo de transmisión de Trypanosoma cruzi por transfusión de sangre en México Risk of Trypanosoma cruzi transmission by blood transfusion in Mexico

    Directory of Open Access Journals (Sweden)

    Carmen Guzmán Bracho

    1998-08-01

    Full Text Available Datos de finales de los años ochenta indican que 1,6% de la población mexicana estaba infectada por la enfermedad de Chagas y que la transmisión de Trypanosoma cruzi por transfusión de sangre ocurría en casi todos los estados, si bien en zonas de diversa extensión. El riesgo de transmisión por esa vía está poco documentado en México, por lo que en 1994 se realizó una encuesta centinela de 18 bancos de sangre de la Secretaría de Salud, situados en sendos estados. El estudio tuvo como objeto conocer el riesgo de transmisión por transfusión de sangre y estimar la prevalencia nacional de infección en los candidatos a donantes, para disponer de indicadores generales de la situación actual de la enfermedad y de la relevancia de ese tipo de transmisión. La selección de participantes se basó en criterios operativos: todos los centros estatales de transfusión que contaban con la capacidad para tamizar a los donantes de sangre por lo menos durante un año y los candidatos a donar (n = 64969 que cumplían con los requisitos exigidos por la Norma Oficial Mexicana para la disposición de sangre humana y derivados con fines terapéuticos. Para el análisis de los resultados, los centros se agruparon según el flujo migratorio para detectar cualquier posible relación entre este y la transmisión de la enfermedad de Chagas en el país. Como prueba de tamizaje se usó la hemaglutinación indirecta con reactivo producido por el Instituto Nacional de Diagnóstico y Referencia Epidemiológicos y donado a los bancos de sangre. Los casos positivos se confirmaron mediante la inmunofluorescencia indirecta. Se detectaron 996 personas con resultados positivos, que representan una prevalencia de 1,5% (IC95%: 1,44 a 1,63. La concordancia de los resultados finales entre los laboratorios locales y el laboratorio central presentó un índice kappa de 0,87 (IC95%: 0,862 a 0,877. En las ciudades con los índices más altos de emigración el riesgo de

  12. GENETIC ANALYSIS OF YIELD AND YIELD COMPONENTS IN ...

    African Journals Online (AJOL)

    ACSS

    2017-11-16

    Nov 16, 2017 ... used different genotypes and the environmental conditions under which their ... and Jinks (1971):. Y = m + aa + βd + a2aa + 2aβad +β2dd … .... /plant, 100-grain weight per plant and Grain yield per plant (g) of six generations in IET6279 X IR70445-146-3-. 3 cross. Traits. Generation. Mean. Standard. Range.

  13. Potato yield and yield structure depending on irrigation

    Directory of Open Access Journals (Sweden)

    Milić Stanko

    2010-01-01

    Full Text Available In the agroclimatic conditions of the Vojvodina Province, the application of an economic water regime and modern technology is necessary for stable and intensive potato production. A two-year experiment on calcareous chernozem was carried out to determine how irrigation and different pre-irrigation soil moisture affect potato yield and distribution of tuber fraction in the potato yield. The block-design trial had four replicates and was adapted for sprinkler irrigation conditions. It included four treatments: irrigation with pre-irrigation moisture levels of 60 % of field water capacity (FC, irrigation with pre-irrigation moisture levels of 70 % (FC, irrigation with pre-irrigation moisture levels of 80% (FC, and a non-irrigated control treatment. Irrigation significantly increased the yield of potato, which increased from 37.27 % to 75.86 %. Under irrigation, the percentage of small fractions decreased in favour of the 55 mm one, or fractions above the 45-55 mm range. On average, irrigated treatments produced significantly more tubers than the conditions of natural water supply. .

  14. Differential Editosome Protein Function between Life Cycle Stages of Trypanosoma brucei.

    Science.gov (United States)

    McDermott, Suzanne M; Guo, Xuemin; Carnes, Jason; Stuart, Kenneth

    2015-10-09

    Uridine insertion and deletion RNA editing generates functional mitochondrial mRNAs in Trypanosoma brucei. The mRNAs are differentially edited in bloodstream form (BF) and procyclic form (PF) life cycle stages, and this correlates with the differential utilization of glycolysis and oxidative phosphorylation between the stages. The mechanism that controls this differential editing is unknown. Editing is catalyzed by multiprotein ∼20S editosomes that contain endonuclease, 3'-terminal uridylyltransferase, exonuclease, and ligase activities. These editosomes also contain KREPB5 and KREPA3 proteins, which have no functional catalytic motifs, but they are essential for parasite viability, editing, and editosome integrity in BF cells. We show here that repression of KREPB5 or KREPA3 is also lethal in PF, but the effects on editosome structure differ from those in BF. In addition, we found that point mutations in KREPB5 or KREPA3 differentially affect cell growth, editosome integrity, and RNA editing between BF and PF stages. These results indicate that the functions of KREPB5 and KREPA3 editosome proteins are adjusted between the life cycle stages. This implies that these proteins are involved in the processes that control differential editing and that the 20S editosomes differ between the life cycle stages. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  15. Bioenergetic profiling of Trypanosoma cruzi life stages using Seahorse extracellular flux technology.

    Science.gov (United States)

    Shah-Simpson, Sheena; Pereira, Camila F A; Dumoulin, Peter C; Caradonna, Kacey L; Burleigh, Barbara A

    2016-08-01

    Energy metabolism is an attractive target for the development of new therapeutics against protozoan pathogens, including Trypanosoma cruzi, the causative agent of human Chagas disease. Despite emerging evidence that mitochondrial electron transport is essential for the growth of intracellular T. cruzi amastigotes in mammalian cells, fundamental knowledge of mitochondrial energy metabolism in this parasite life stage remains incomplete. The Clark-type electrode, which measures the rate of oxygen consumption, has served as the traditional tool to study mitochondrial energetics and has contributed to our understanding of it in T. cruzi. Here, we evaluate the Seahorse XF(e)24 extracellular flux platform as an alternative method to assess mitochondrial bioenergetics in isolated T. cruzi parasites. We report optimized assay conditions used to perform mitochondrial stress tests with replicative life cycle stages of T. cruzi using the XF(e)24 instrument, and discuss the advantages and potential limitations of this methodology, as applied to T. cruzi and other trypanosomatids. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Isothermal microcalorimetry, a new tool to monitor drug action against Trypanosoma brucei and Plasmodium falciparum.

    Directory of Open Access Journals (Sweden)

    Tanja Wenzler

    Full Text Available Isothermal microcalorimetry is an established tool to measure heat flow of physical, chemical or biological processes. The metabolism of viable cells produces heat, and if sufficient cells are present, their heat production can be assessed by this method. In this study, we investigated the heat flow of two medically important protozoans, Trypanosoma brucei rhodesiense and Plasmodium falciparum. Heat flow signals obtained for these pathogens allowed us to monitor parasite growth on a real-time basis as the signals correlated with the number of viable cells. To showcase the potential of microcalorimetry for measuring drug action on pathogenic organisms, we tested the method with three antitrypanosomal drugs, melarsoprol, suramin and pentamidine and three antiplasmodial drugs, chloroquine, artemether and dihydroartemisinin, each at two concentrations on the respective parasite. With the real time measurement, inhibition was observed immediately by a reduced heat flow compared to that in untreated control samples. The onset of drug action, the degree of inhibition and the time to death of the parasite culture could conveniently be monitored over several days. Microcalorimetry is a valuable element to be added to the toolbox for drug discovery for protozoal diseases such as human African trypanosomiasis and malaria. The method could probably be adapted to other protozoan parasites, especially those growing extracellularly.

  17. Enhancing effects of gamma interferon on phagocytic cell association with and killing of Trypanosoma cruzi

    Science.gov (United States)

    Wirth, J. J.; Kierszenbaum, F.; Sonnenfeld, G.; Zlotnik, A.

    1985-01-01

    Results are reported from a study of the influence gamma interferon (GIFN) and interleukin 2 (IL2) have on the capability of P388D1 cells and mouse resident peritoneal macrophages (MPM) to attach to the blood-resident parasites Trypanosoma cruzi and kill them. Cultures of trypomastigote forms of the Tulahuen strain of T. cruzi grown in bovine serum were introduced into peritoneal cells of mice, along with P388D1 cells incubated with GIFN, IL2 and both. Control cells were also maintained. Statistical analysis were then performed on data on counts of the number of dead T. Cruzi cells. The GIFN enhanced the interaction of MPM and P388D1 cells with the surface of T. Cruzi, provided the interaction was given over 12 hr to take place. A depression of the cytotoxicity of P388D1 cells was attributed to mediation by H2O2, an effect partially offset by incubation with the lymphokine GIFN.

  18. Protein preparation, crystallization and preliminary X-ray analysis of Trypanosoma cruzi nucleoside diphosphate kinase 1

    International Nuclear Information System (INIS)

    Gómez Barroso, J. A.; Pereira, H.; Miranda, M.; Pereira, C.; Garratt, R. C.; Aguilar, C. F.

    2010-01-01

    T. cruzi TcNDPK1 was overexpressed in Escherichia coli as an N-terminally poly-His-tagged fusion protein and crystallized. The flagellated protozoan parasite Trypanosoma cruzi is the aetiological agent of Chagas disease. Nucleoside diphosphate kinases (NDPKs) are enzymes that are involved in energy management and nucleoside balance in the cell. T. cruzi TcNDPK1, a canonical isoform, was overexpressed in Escherichia coli as an N-terminally poly-His-tagged fusion protein and crystallized. Crystals grew after 72 h in 0.2 M MgCl 2 , 20% PEG 3350. Data were collected to 3.5 Å resolution using synchrotron X-ray radiation at the National Synchrotron Light Laboratory (Campinas, Brazil). The crystals belonged to the trigonal space group P3, with unit-cell parameters a = b = 127.84, c = 275.49 Å. Structure determination is under way and will provide relevant information that may lead to the first step in rational drug design for the treatment of Chagas disease

  19. Differential expression profiles in the midgut of Triatoma infestans infected with Trypanosoma cruzi.

    Directory of Open Access Journals (Sweden)

    Diego S Buarque

    Full Text Available Chagas disease, or American trypanosomiasis, is a parasitic disease caused by the protozoan Trypanosoma cruzi and is transmitted by insects from the Triatominae subfamily. To identify components involved in the protozoan-vector relationship, we constructed and analyzed cDNA libraries from RNA isolated from the midguts of uninfected and T. cruzi-infected Triatoma infestans, which are major vectors of Chagas disease. We generated approximately 440 high-quality Expressed Sequence Tags (ESTs from each T. infestans midgut cDNA library. The sequences were grouped in 380 clusters, representing an average length of 664.78 base pairs (bp. Many clusters were not classified functionally, representing unknown transcripts. Several transcripts involved in different processes (e.g., detoxification showed differential expression in response to T. cruzi infection. Lysozyme, cathepsin D, a nitrophorin-like protein and a putative 14 kDa protein were significantly upregulated upon infection, whereas thioredoxin reductase was downregulated. In addition, we identified several transcripts related to metabolic processes or immunity with unchanged expressions, including infestin, lipocalins and defensins. We also detected ESTs encoding juvenile hormone binding protein (JHBP, which seems to be involved in insect development and could be a target in control strategies for the vector. This work demonstrates differential gene expression upon T. cruzi infection in the midgut of T. infestans. These data expand the current knowledge regarding vector-parasite interactions for Chagas disease.

  20. Usefulness of microsatellite typing in population genetic studies of Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Macedo Andrea M

    2001-01-01

    Full Text Available Through microsatellite analysis of 53 monoclonal populations of Trypanosoma cruzi, we found a remarkable degree of genetic polymorphism with no single multilocus genotype being observed more than once. The microsatellite profile proved to be stable during 70 generations of the CL Brener clone in culture. The microsatellite profiling presented also high diagnostic sensitivity since DNA amplifications could be achieved with less than 100 fg DNA, corresponding to half parasite total DNA content. Based on these technical attributes the microsatellite assay turns out to be an important tool for direct typing T. cruzi in biological samples. By using this approach we were able to type T. cruzi in feces of artificially infected bugs and in single cells sorted by FACS. The microsatellites have shown to be excellent markers for T. cruzi phylogenetic reconstruction. We used maximum parsimony based on the minimum number of mutational steps to build an unrooted Wagner network, which confirms previous conclusions based on the analysis of the D7 domain of the LSU rDNA gene that T. cruzi is composed by two major groups. We also obtained evidence that strains belonging to rRNA group 2 are subdivided into two genetically distant clusters, and that one of these clusters is more related to rRNA group 1/2. These results suggest different origins for these strains.

  1. Trypanosoma cruzi strains from triatomine collected in Bahia and Rio Grande do Sul, Brazil

    Directory of Open Access Journals (Sweden)

    Aline Rimoldi Ribeiro

    2014-04-01

    Full Text Available OBJECTIVE Collection of triatomines in domestic, peridomestic and sylvatic environments in states of Bahia and Rio Grande do Sul, Northeastern and Southern Brazil respectively, and isolation of Trypanosoma cruzi strains. METHODS First, the captured triatomines were identified using insect identification keys, then their intestinal content was examined by abdominal compression, and the samples containing trypanosomatid forms were inoculated in LIT medium and Swiss mice. RESULTS Six triatomine species were collected in cities in Bahia, namely Panstrongylus geniculatus (01, Triatoma melanocephala (11, T. lenti (94, T. pseudomaculata (02, T. sherlocki (26 and T. sordida (460, and two in cities in Rio Grande do Sul, namely T. circummaculata (11 and T. rubrovaria (115. Out of the specimens examined, T. cruzi was isolated from 28 triatomine divided into four different species: T. melanocephala (one, T. lenti (one, T. rubrovaria (16 and T. sordida (10. Their index of natural infection by T. cruzi was 6.4%. CONCLUSIONS The isolation of T. cruzi strains from triatomines found in domestic and peridomestic areas shows the potential risk of transmission of Chagas disease in the studied cities. The maintenance of those T. cruzi strains in laboratory is intended to promote studies that facilitate the understanding of the parasite-vector-host relationship.

  2. Benznidazole biotransformation and multiple targets in Trypanosoma cruzi revealed by metabolomics.

    Directory of Open Access Journals (Sweden)

    Andrea Trochine

    2014-05-01

    Full Text Available The first line treatment for Chagas disease, a neglected tropical disease caused by the protozoan parasite Trypanosoma cruzi, involves administration of benznidazole (Bzn. Bzn is a 2-nitroimidazole pro-drug which requires nitroreduction to become active, although its mode of action is not fully understood. In the present work we used a non-targeted MS-based metabolomics approach to study the metabolic response of T. cruzi to Bzn.Parasites treated with Bzn were minimally altered compared to untreated trypanosomes, although the redox active thiols trypanothione, homotrypanothione and cysteine were significantly diminished in abundance post-treatment. In addition, multiple Bzn-derived metabolites were detected after treatment. These metabolites included reduction products, fragments and covalent adducts of reduced Bzn linked to each of the major low molecular weight thiols: trypanothione, glutathione, γ-glutamylcysteine, glutathionylspermidine, cysteine and ovothiol A. Bzn products known to be generated in vitro by the unusual trypanosomal nitroreductase, TcNTRI, were found within the parasites, but low molecular weight adducts of glyoxal, a proposed toxic end-product of NTRI Bzn metabolism, were not detected.Our data is indicative of a major role of the thiol binding capacity of Bzn reduction products in the mechanism of Bzn toxicity against T. cruzi.

  3. Structure of a Trypanosoma brucei α/β-hydrolase fold protein with unknown function

    International Nuclear Information System (INIS)

    Merritt, Ethan A.; Holmes, Margaret; Buckner, Frederick S.; Van Voorhis, Wesley C.; Quartly, Erin; Phizicky, Eric M.; Lauricella, Angela; Luft, Joseph; DeTitta, George; Neely, Helen; Zucker, Frank; Hol, Wim G. J.

    2008-01-01

    T. brucei gene Tb10.6k15.0140 codes for an α/β-hydrolase fold protein of unknown function. The 2.2 Å crystal structure shows that members of this sequence family retain a conserved Ser residue at the expected site of a catalytic nucleophile, but that trypanosomatid sequences lack structural homologs for the other expected residues of the catalytic triad. The structure of a structural genomics target protein, Tbru020260AAA from Trypanosoma brucei, has been determined to a resolution of 2.2 Å using multiple-wavelength anomalous diffraction at the Se K edge. This protein belongs to Pfam sequence family PF08538 and is only distantly related to previously studied members of the α/β-hydrolase fold family. Structural superposition onto representative α/β-hydrolase fold proteins of known function indicates that a possible catalytic nucleophile, Ser116 in the T. brucei protein, lies at the expected location. However, the present structure and by extension the other trypanosomatid members of this sequence family have neither sequence nor structural similarity at the location of other active-site residues typical for proteins with this fold. Together with the presence of an additional domain between strands β6 and β7 that is conserved in trypanosomatid genomes, this suggests that the function of these homologs has diverged from other members of the fold family

  4. Interaction between Didelphis albiventris and Triatoma infestans in relation to Trypanosoma cruzi transmission

    Directory of Open Access Journals (Sweden)

    Nicolás J. Schweigmann

    1995-12-01

    Full Text Available This paper attempts to prove if a high Trypanosoma cruzi prevalence of opossums might be reached with few potential infective contacts. One non-infected Didelphis albiventris to T. cruzi and 10 infected nymphs of Triatoma infestans were left together during 23 hr in a device that simulated a natural opossum burrow. Twenty-six replicates were perfomed using marsupials and triatomines only once. Potentially infective contacts occurred in all the trials. From the 26 opossums used in trials, 54% did not eat any bug. Of the 260 bugs used, 21% were predated. In the 25 trials involving 205 surving bugs, 36 % of them did not feed. In 15/25 cases, maior ou igual a 60% of the triatomines were able to feed. The parasitological follow-up of 24 opossums showed that among 10 that had eaten bugs, 4 turned out infected and among the 14 that had not predate, 3 (21% became positive. In sum, 7/24 (29% of the marsupials acquired the infection after the experiment. This infection rate was similar to the prevalences found for the opossum population of Santiago del Estero, Argentina, suggesting that the prevalences observed in the field might be reached if each marsupial would encounter infected bugs just once in its lifetime.

  5. Studying nanotoxic effects of CdTe quantum dots in Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Cecilia Stahl Vieira

    2011-03-01

    Full Text Available Semiconductor nanoparticles, such as quantum dots (QDs, were used to carry out experiments in vivo and ex vivo with Trypanosoma cruzi. However, questions have been raised regarding the nanotoxicity of QDs in living cells, microorganisms, tissues and whole animals. The objective of this paper was to conduct a QD nanotoxicity study on living T. cruzi protozoa using analytical methods. This was accomplished using in vitro experiments to test the interference of the QDs on parasite development, morphology and viability. Our results show that after 72 h, a 200 μM cadmium telluride (CdTe QD solution induced important morphological alterations in T. cruzi, such as DNA damage, plasma membrane blebbing and mitochondrial swelling. Flow cytometry assays showed no damage to the plasma membrane when incubated with 200 μM CdTe QDs for up to 72 h (propidium iodide cells, giving no evidence of classical necrosis. Parasites incubated with 2 μM CdTe QDs still proliferated after seven days. In summary, a low concentration of CdTe QDs (2 μM is optimal for bioimaging, whereas a high concentration (200 μM CdTe could be toxic to cells. Taken together, our data indicate that 2 μM QD can be used for the successful long-term study of the parasite-vector interaction in real time.

  6. Flux Analysis of the Trypanosoma brucei Glycolysis Based on a Multiobjective-Criteria Bioinformatic Approach

    Directory of Open Access Journals (Sweden)

    Amine Ghozlane

    2012-01-01

    Full Text Available Trypanosoma brucei is a protozoan parasite of major of interest in discovering new genes for drug targets. This parasite alternates its life cycle between the mammal host(s (bloodstream form and the insect vector (procyclic form, with two divergent glucose metabolism amenable to in vitro culture. While the metabolic network of the bloodstream forms has been well characterized, the flux distribution between the different branches of the glucose metabolic network in the procyclic form has not been addressed so far. We present a computational analysis (called Metaboflux that exploits the metabolic topology of the procyclic form, and allows the incorporation of multipurpose experimental data to increase the biological relevance of the model. The alternatives resulting from the structural complexity of networks are formulated as an optimization problem solved by a metaheuristic where experimental data are modeled in a multiobjective function. Our results show that the current metabolic model is in agreement with experimental data and confirms the observed high metabolic flexibility of glucose metabolism. In addition, Metaboflux offers a rational explanation for the high flexibility in the ratio between final products from glucose metabolism, thsat is, flux redistribution through the malic enzyme steps.

  7. Domestic Pig (Sus scrofa) as an Animal Model for Experimental Trypanosoma cruzi Infection

    Science.gov (United States)

    Yauri, Verónica; Castro-Sesquen, Yagahira E.; Verastegui, Manuela; Angulo, Noelia; Recuenco, Fernando; Cabello, Ines; Malaga, Edith; Bern, Caryn; Gavidia, Cesar M.; Gilman, Robert H.

    2016-01-01

    Pigs were infected with a Bolivian strain of Trypanosoma cruzi (genotype I) and evaluated up to 150 days postinoculation (dpi) to determine the use of pigs as an animal model of Chagas disease. Parasitemia was observed in the infected pigs during the acute phase (15–40 dpi). Anti-T.cruzi immunoglobulin M was detected during 15–75 dpi; high levels of anti-T.cruzi immunoglobulin G were detected in all infected pigs from 75 to 150 dpi. Parasitic DNA was observed by western blot (58%, 28/48) and polymerase chain reaction (27%, 13/48) in urine samples, and in the brain (75%, 3/4), spleen (50%, 2/4), and duodenum (25%, 1/4), but no parasitic DNA was found in the heart, colon, and kidney. Parasites were not observed microscopically in tissues samples, but mild inflammation, vasculitis, and congestion was observed in heart, brain, kidney, and spleen. This pig model was useful for the standardization of the urine test because of the higher volume that can be obtained as compared with other small animal models. However, further experiments are required to observe pathological changes characteristic of Chagas disease in humans. PMID:26928841

  8. Potential Role of Carvedilol in the Cardiac Immune Response Induced by Experimental Infection with Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Aline Luciano Horta

    2017-01-01

    Full Text Available Trypanosoma cruzi causes a cardiac infection characterized by an inflammatory imbalance that could become the inciting factor of the illness. To this end, we evaluated the role of carvedilol, a beta-blocker with potential immunomodulatory properties, on the immune response in C57BL/6 mice infected with VL-10 strain of T. cruzi in the acute phase. Animals (n=40 were grouped: (i not infected, (ii infected, (iii infected + carvedilol, and (iv not infected + carvedilol. We analyzed parameters related to parasitemia, plasma levels of TNF, IL-10, and CCL2, and cardiac histopathology after the administration of carvedilol for 30 days. We did not observe differences in the maximum peaks of parasitemia in the day of their detection among the groups. The plasma TNF was elevated at 60 days of infection in mice treated or not with carvedilol. However, we observed a decreased CCL2 level and increased IL-10 levels in those infected animals treated with carvedilol, which impacted the reduction of the inflammatory infiltration in cardiac tissue. For this experimental model, carvedilol therapy was not able to alter the levels of circulating parasites but modulates the pattern of CCL2 and IL-10 mediators when the VL10 strain of T. cruzi was used in C57BL6 mice.

  9. Influence of environmental enrichment on the behavior and physiology of mice infected by Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Déborah Maria Moreira da Silva

    Full Text Available Abstract INTRODUCTION: Enriched environments normally increase behavioral repertoires and diminish the expression of abnormal behaviors and stress-related physiological problems in animals. Although it has been shown that experimental animals infected with microorganisms can modify their behaviors and physiology, few studies have evaluated how environmental enrichment affects these parameters. This study aimed to evaluate the effects of environmental enrichment on the behavior and physiology of confined mice infected with Trypanosoma cruzi. METHODS: The behaviors of 20 T. cruzi-infected mice and 20 non-infected mice were recorded during three treatments: baseline, enrichment, and post-enrichment. Behavioral data were collected using scan sampling with instantaneous recording of behavior every 30s, totaling 360h. Plasma TNF, CCL2, and IL-10 levels and parasitemia were also evaluated in infected enriched/non-enriched mice. Behavioral data were evaluated by Friedman’s test and physiological data by one-way ANOVA and area under the curve (AUC analysis. RESULTS: Results showed that environmental enrichment significantly increased exploratory behaviors and diminished inactivity. The use of environmental enrichment did not diminish circulating levels of TNF and IL-10 but diminished circulating levels of CCL2 and parasitemia. CONCLUSIONS: Positive behavioral and physiological effects of environmental enrichment were observed in mice living in enriched cages. Thus, environmental enrichment improved the welfare of these animals.

  10. Risk factors associated with Trypanosoma cruzi exposure in domestic dogs from a rural community in Panama.

    Science.gov (United States)

    Saldaña, Azael; Calzada, José E; Pineda, Vanessa; Perea, Milixa; Rigg, Chystrie; González, Kadir; Santamaria, Ana Maria; Gottdenker, Nicole L; Chaves, Luis F

    2015-11-01

    Chagas disease, caused by Trypanosoma cruzi infection, is a zoonosis of humans, wild and domestic mammals, including dogs. In Panama, the main T. cruzi vector is Rhodnius pallescens, a triatomine bug whose main natural habitat is the royal palm, Attalea butyracea. In this paper, we present results from three T. cruzi serological tests (immunochromatographic dipstick, indirect immunofluorescence and ELISA) performed in 51 dogs from 24 houses in Trinidad de Las Minas, western Panama. We found that nine dogs were seropositive (17.6% prevalence). Dogs were 1.6 times more likely to become T. cruzi seropositive with each year of age and 11.6 times if royal palms where present in the peridomiciliary area of the dog's household or its two nearest neighbours. Mouse-baited-adhesive traps were employed to evaluate 12 peridomestic royal palms. All palms were found infested with R. pallescens with an average of 25.50 triatomines captured per palm. Of 35 adult bugs analysed, 88.6% showed protozoa flagellates in their intestinal contents. In addition, dogs were five times more likely to be infected by the presence of an additional domestic animal species in the dog's peridomiciliary environment. Our results suggest that interventions focused on royal palms might reduce the exposure to T. cruzi infection.

  11. Is the anti-tumor property of Trypanosoma cruzi infection mediated by its Calreticulin?

    Directory of Open Access Journals (Sweden)

    Galia Andrea Ramírez-Toloza

    2016-07-01

    Full Text Available Eight to 10 million people in 21 endemic countries are infected with Trypanosoma cruzi. However, only 30% of those infected develop symptoms of Chagas’ disease, a chronic, neglected tropical disease worldwide. Similar to other pathogens, T. cruzi has evolved to resist the host immune response. Studies, performed 80 years ago in the Soviet Union, proposed that T. cruzi infects tumor cells with similar capacity to that displayed for target tissues such as cardiac, aortic or digestive. An antagonistic relationship between T. cruzi infection and cancer development was also proposed, but the molecular mechanisms involved have remained largely unknown. Probably, a variety of T. cruzi molecules is involved. This review focuses on how T. cruzi calreticulin (TcCRT, exteriorized from the endoplasmic reticulum, targets the first classical complement component C1 and negatively regulates the Classical Complement activation cascade, promoting parasite infectivity. We propose that this C1-dependent TcCRT-mediated virulence is critical to explain, at least an important part, of the parasite capacity to inhibit tumor development. We will discuss how TcCRT, by directly interacting with venous and arterial endothelial cells, inhibits angiogenesis and tumor growth. Thus, these TcCRT functions not only illustrate T. cruzi interactions with the host immune defensive strategies, but also illustrate a possible co-evolutionary adaptation to privilege a prolonged interaction with its host.

  12. Morphometry of submucous and myenteric esophagic plexus of dogs experimentally reinfected with Trypanosoma cruzi

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    Machado Evandro MM

    2001-01-01

    Full Text Available We carried out a morphometric study of the esophagus of cross-bred dogs experimentally infected or consecutively reinfected with Trypanosoma cruzi 147 and SC-1 strains, in order to verify denervation and/or neuronal hypertrophy in the intramural plexus. The animals were sacrificed in the chronic stage, 38 months after the initial infection. Neither nests of amastigotes, nor myositis or ganglionitis, were observed in all third inferior portions of esophageal rings analyzed. No nerve cell was identified in the submucous of this organ. There was no significant difference (p>0.05 between the number, maximum diameter, perimeter, or area and volume of the nerve cells of the myenteric plexus of infected and/or reinfected dogs and of the non-infected ones. In view of these results we may conclude that the 147 and SC-1 strains have little neurotropism and do not determine denervation and/or hypertrophy in the intramural esophageal plexuses in the animals studied, independent of the reinfections.

  13. Decay-accelerating factor 1 deficiency exacerbates Trypanosoma cruzi-induced murine chronic myositis.

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    Solana, María E; Ferrer, María F; Novoa, María Mercedes; Song, Wen-Chao; Gómez, Ricardo M

    2012-10-01

    Murine infection with Trypanosoma cruzi (Tc) has been used to study the role of T-cells in the pathogenesis of human inflammatory idiopathic myositis. Absence of decay-accelerating factor 1 (Daf1) has been shown to enhance murine T-cell responses and autoimmunity. To determine whether Daf1 deficiency can exacerbate Tc-induced myositis, C57BL/6 DAF(+/+) and DAF(-/-) mice were inoculated with 5 × 10(4) trypomastigotes, and their morbidity, parasitemia, parasite burden, histopathology, and T-cell expansion were studied in the acute and chronic stages. DAF(-/-) mice had lower parasitemia and parasite burden but higher morbidity, muscle histopathology, and increased number of CD44(+) (activated/memory phenotype) splenic CD4(+) and CD8(+) T-cells. An enhanced CD8(+) T-cell immune-specific response may explain the lower parasitemia and parasite burden levels and the increase in histopathological lesions. We propose that Tc-inoculated DAF(-/-) mice are a useful model to study T-cell mediated immunity in skeletal muscle tissues. Copyright © 2012 Wiley Periodicals, Inc.

  14. Effects of buthionine sulfoximine nifurtimox and benznidazole upon trypanothione and metallothionein proteins in Trypanosoma cruzi.

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    JUAN DIEGO MAYA

    2004-01-01

    Full Text Available Proteins rich in sulfhydryl groups, such as metallothionein, are present in several strains of the parasite Trypanosoma cruzi, the etiological agent of Chagas' disease. Metallothionein-like protein concentrations ranged from 5.1 to 13.2 pmol/mg protein depending on the parasite strain and growth phase. Nifurtimox and benznidazole, used in the treatment of Chagas' disease, decreased metallothionein activity by approximately 70%. T. cruzi metallothionein was induced by ZnCl2. Metallothionein from T. cruzi was partially purified and its monobromobimane derivative showed a molecular weight of approximately 10,000 Da by SDS-PAGE analysis. The concentration of trypanothione, the major glutathione conjugate in T. cruzi, ranged from 3.8 to 10.8 nmol/mg protein, depending on the culture phase. The addition of buthionine sulfoximine to the protozoal culture considerably reduced the concentration of trypanothione and had no effect upon the metallothionein concentration. The possible contribution of metallothionein-like proteins to drug resistance in T. cruzi is discussed.

  15. [Entomological study of Trypanosoma cruzi vectors in the rural communities of Sucre state, Venezuela].

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    García-Jordán, Noris; Berrizbeitia, Mariolga; Concepción, Juan Luis; Aldana, Elis; Cáceres, Ana; Quiñones, Wilfredo

    2015-01-01

    The ecological niche of Reduvidae vectors has been modified due to environmental changes and human encroachment into the rural areas. This study evaluates the current entomological indices of triatomines responsible for Trypanosoma cruzi infection in Sucre State, Venezuela. A cross-sectional and prospective study was conducted in 95 towns and 577 dwellings in the 15 municipalities of the state of Sucre, Venezuela, from August to November, 2008. Triatomine bugs were identified on the basis of morphological characteristics, and their feces examined for T. cruzi infection through direct microscopy. Positive slides were stained with Giemsa and parasites were identified by morphologic characterization. The entomological indices expressing the highest values were dispersion (16.67%) and household colonization (33.33%). The triatomine species captured were: Rhodnius prolixus , Rhodnius main intradomiciliary vector. Despite the low index of vector infection (1.72%), the existence of species with domiciliary and peridomiciliary reproductive success ensures the persistence of the epidemiological chain both for the disease and the parasite.

  16. Design of Trypanosoma rangeli sialidase mutants with improved trans-sialidase activity.

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    Christian Nyffenegger

    Full Text Available A sialidase (EC 3.2.1.18 from the non-pathogenic Trypanosoma rangeli, TrSA, has been shown to exert trans-sialidase activity after mutation of five specific amino acids in the active site (M96V, A98P, S120Y, G249Y, Q284P to form the so-called TrSA5mut enzyme. By computational and hypothesis driven approaches additional mutations enhancing the trans-sialidase activity have been suggested. In the present work, we made a systematic combination of these mutations leading to seven new variants of the T. rangeli sialidase, having 6-16 targeted amino acid mutations. The resulting enzyme variants were analyzed via kinetics for their ability to carry out trans-sialidase reaction using CGMP and D-lactose as substrates. The sialidase variants with 15 and 16 mutations, respectively, exhibited significantly improved trans-sialidase activity for D-lactose sialylation. Our results corroborate, that computational studies of trans-glycosylation can be a valuable input in the design of novel trans-glycosidases, but also highlight the importance of experimental validation in order to assess the performance. In conclusion, two of the seven mutants displayed a dramatic switch in specificity from hydrolysis towards trans-sialylation and constitute the most potent trans-sialidase mutants of TrSA described in literature to date.

  17. Coadministration of cruzipain and GM-CSF DNAs, a new immunotherapeutic vaccine against Trypanosoma cruzi infection.

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    Cerny, Natacha; Sánchez Alberti, Andrés; Bivona, Augusto E; De Marzi, Mauricio C; Frank, Fernanda M; Cazorla, Silvia I; Malchiodi, Emilio L

    2016-01-01

    Therapeutic vaccine research and development are especially important in Chagas disease considering the characteristics of the chronic infection and the number of people in the Americas living with a parasite infection for decades. We have previously reported the efficacy of attenuated Salmonella enterica (S) carrying plasmid encoding cruzipain (SCz) to protect against Trypanosoma cruzi infection. In the present work we investigated whether Cz DNA vaccine immunotherapy could be effective in controlling an ongoing T. cruzi infection in mice. We here report the intramuscular administration of naked Cz DNA or the oral administration of Salmonella as Cz DNA delivery system as therapeutic vaccines in mice during acute or chronic infection. The coadministration of a plasmid encoding GM-CSF improved vaccine performance, indicating that the stimulation of innate immune cells is needed in the event of an ongoing infection. These therapeutic vaccines were able to address the response to a protective and sustained Th1 biased profile not only against Cz but also against a variety of parasite antigens. The combined therapeutic vaccine during the chronic phase of infection prevents tissue pathology as shown by a reduced level of enzyme activity characteristic of tissue damage and a tissue status compatible with normal tissue. The obtained results suggest that immunotherapy with Cz and GM-CSF DNAs, either alone or in combination with other drug treatments, may represent a promising alternative for Chagas disease therapy.

  18. The Acute Phase of Trypanosoma cruzi Infection Is Attenuated in 5-Lipoxygenase-Deficient Mice

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    Adriana M. C. Canavaci

    2014-01-01

    Full Text Available In the present work we examine the contribution of 5-lipoxygenase- (5-LO- derived lipid mediators to immune responses during the acute phase of Trypanosoma cruzi infection in 5-LO gene knockout (5-LO−/− mice and wild-type (WT mice. Compared with WT mice, the 5-LO−/− mice developed less parasitemia/tissue parasitism, less inflammatory cell infiltrates, and a lower mortality. This resistance of 5-LO−/− mice correlated with several differences in the immune response to infection, including reduced PGE2 synthesis; sustained capacity of splenocytes to produce high levels of interleukin (IL-12 early in the infection; enhanced splenocyte production of IL-1β, IL-6, and IFN-γ; rapid T-cell polarization to secrete high quantities of IFN-γ and low quantities of IL-10; and greater numbers of CD8+CD44highCD62Llow memory effector T cells at the end of the acute phase of infection. The high mortality in WT mice was associated with increased production of LTB4/LTC4, T cell bias to produce IFN-γ, high levels of serum nitrite, and marked protein extravasation into the peritoneal cavity, although survival was improved by treatment with a cys-LT receptor 1 antagonist. These data also provide evidence that 5-LO-derived mediators negatively affect host survival during the acute phase of T. cruzi infection.

  19. Standardization of serological tests for detecting anti-Trypanosoma cruzi antibodies in dogs

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    M. A. Lauricella

    1993-09-01

    Full Text Available This paper reports on the standardization of four serological reactions currently used in human serodiagnosis for the detection of anti-Trypanosoma cruzi antibodies in naturally and experimentally infected dogs. Indirect immunofluorescence test (IFAT and hemagglutination test (IHAT were standardized, and complement fixation test (CFT and direct agglutination test (DAT were used for diagnostic confirmation. Four hundred and eighty one mongrel dogs that were studied by xenodiagnosis were used: (1 parasitemic dogs of two localities of endemic area (EA of Santiago del Estero province in Argentina (n = 134; (2 non-parasitemic dogs of the same area (n = 285; (3 dogs experimentally infected with T. cruzi in the patent period (n = 6; (4 non-infected dogs (n = 56 which were born in the city of Buenos Aires (BA, one non-EA for Chagas' disease. For IFAT, parasitemic dogs EA showed 95% of reactive sera. Non parasitemic dogs EA showed 77% of non reactive sera. None sera from BA were reactive for dilutions higher than four. For IHAT, 84% of sera of parasitemic dogs EA showed serological reactivity and among non parasitemic dogs BA, 61% were non reactive, while the remainder showed at most titres of 1/16. The cut-off titres for IFAT and IHAT were 1/16 and 1/32 respectively, and for CFT and DAT 1/1 and 1/128 respectively. Sensitivity for IFAT, IHAT, CF and DAT were 95%, 84%, 97% and 95% respectively.

  20. Identification and characterization of a stage specific membrane protein involved in flagellar attachment in Trypanosoma brucei.

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    Katherine Woods

    Full Text Available Flagellar attachment is a visibly striking morphological feature of African trypanosomes but little is known about the requirements for attachment at a molecular level. This study characterizes a previously undescribed membrane protein, FLA3, which plays an essential role in flagellar attachment in Trypanosoma brucei. FLA3 is heavily N-glycosylated, locates to the flagellar attachment zone and appears to be a bloodstream stage specific protein. Ablation of the FLA3 mRNA rapidly led to flagellar detachment and a concomitant failure of cytokinesis in the long slender bloodstream form but had no effect on the procyclic form. Flagellar detachment was obvious shortly after induction of the dsRNA and the newly synthesized flagellum was often completely detached after it emerged from the flagellar pocket. Within 12 h most cells possessed detached flagella alongside the existing attached flagellum. These results suggest that proteins involved in attachment are not shared between the new and old attachment zones. In other respects the detached flagella appear normal, they beat rapidly although directional motion was lost, and they possess an apparently normal axoneme and paraflagellar rod structure. The flagellar attachment zone appeared to be disrupted when FLA3 was depleted. Thus, while flagellar attachment is a constitutive feature of the life cycle of trypanosomes, attachment requires stage specific elements at the protein level.

  1. Trypanosoma cruzi: effects of azadirachtin and ecdysone on the dynamic development in Rhodnius prolixus larvae.

    Science.gov (United States)

    Cortez, M R; Provençano, A; Silva, C E; Mello, C B; Zimmermann, L T; Schaub, G A; Garcia, E S; Azambuja, P; Gonzalez, M S

    2012-07-01

    The effects of azadirachtin and ecdysone on the Trypanosoma cruzi population in the Rhodnius prolixus gut were investigated. T. cruzi were rarely found in the gut compartments of azadirachtin-treated larvae. High parasite numbers were observed in the stomach of the control and ecdysone groups until 10 days after treatment and in the small intestine and rectum until 25 days after treatment. High percentages of round forms developed in the stomachs of all groups, whereas azadirachtin blocked the development of protozoan intermediate forms. This effect was counteracted by ecdysone therapy. In the small intestine and rectum, epimastigotes predominated for all groups, but more of their intermediates developed in the control and ecdysone groups. Azadirachtin supported the development of round forms and their intermediates into trypomastigotes. In the rectum, trypomastigotes did not develop in the azadirachtin group and developed much later after ecdysone therapy. The parallel between the effects of azadirachtin and ecdysone on the host and parasite development is discussed on the basis of the present results because ecdysone appears to act directly or indirectly in determining the synchronic development of T. cruzi forms from round to epimastigotes, but not metacyclic trypomastigotes, in the invertebrate vector. Copyright © 2012 Elsevier Inc. All rights reserved.

  2. Limited antigenic variation in the Trypanosoma cruzi candidate vaccine antigen TSA-1.

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    Knight, J M; Zingales, B; Bottazzi, M E; Hotez, P; Zhan, B

    2014-12-01

    Chagas disease (American trypanosomiasis caused by Trypanosoma cruzi) is one of the most important neglected tropical diseases in the Western Hemisphere. The toxicities and limited efficacies of current antitrypanosomal drugs have prompted a search for alternative technologies such as a therapeutic vaccine comprised of T. cruzi antigens, including a recombinant antigen encoding the N-terminal 65 kDa portion of Trypomastigote surface antigen-1 (TSA-1). With at least six known genetically distinct T. cruzi lineages, variability between the different lineages poses a unique challenge for the development of broadly effective therapeutic vaccine. The variability across the major lineages in the current vaccine candidate antigen TSA-1 has not previously been addressed. To assess the variation in TSA-1, we cloned and sequenced TSA-1 from several different T. cruzi strains representing three of the most clinically relevant lineages. Analysis of the different alleles showed limited variation in TSA-1 across the different strains and fit with the current theory for the evolution of the different lineages. Additionally, minimal variation in known antigenic epitopes for the HLA-A 02 allele suggests that interlineage variation in TSA-1 would not impair the range and efficacy of a vaccine containing TSA-1. © 2014 John Wiley & Sons Ltd.

  3. Trypanosoma (Herpetosoma rangeli Tejera, 1920: intracellular amastigote stages of reproduction in white mice

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    Servio Urdaneta-Morales

    1986-06-01

    Full Text Available The method, site, and stage of multiplication of Trypanosoma (Herpetosoma rangeli Tejera, 1920 has not hitherto been known. "We have now observed many intracellular nests or pseudocysts, containing amastigotes and trypomastigotes of this parasite in the heart, liver, and spleen of suckling (5.0 g male white mice (NMRI strain inoculated i.p. with 9 x 10(4 metatrypomastigotes/g body weight from a 12-day-old culture of the "Dog-82" strain of T. rangeli. At the peak of parasitemia (1.9 x 10(6 trypomastigotes/ml blood, 3 days post-inoculation various tissues were taken for sectioning and staining. The heart was most intensely parasitized. The amastigotes were rounded or ellipsoidal, with a rounded nucleus and the kinetoplast in the form of a straight or curved bar; the average maximum diameter of 50 measured amastigotes was 4.2 p. Binary fission was seen in the nucleus and kinetoplast of some amastigotes; no blood trypomastigotes were seen in division. The above characteristics, as well as the location of the pseudocysts in the tissues, are similar to T. cruzi. Comparison of these results with those reported for other Herpetosoma suggest study of the taxonomic position of T. rangeli.

  4. Myenteric plexus is differentially affected by infection with distinct Trypanosoma cruzi strains in Beagle dogs

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    Nívia Carolina Nogueira-Paiva

    2014-02-01

    Full Text Available Chagasic megaoesophagus and megacolon are characterised by motor abnormalities related to enteric nervous system lesions and their development seems to be related to geographic distribution of distinct Trypanosoma cruzi subpopulations. Beagle dogs were infected with Y or Berenice-78 (Be-78 T. cruzi strains and necropsied during the acute or chronic phase of experimental disease for post mortem histopathological evaluation of the oesophagus and colon. Both strains infected the oesophagus and colon and caused an inflammatory response during the acute phase. In the chronic phase, inflammatory process was observed exclusively in the Be-78 infected animals, possibly due to a parasitism persistent only in this group. Myenteric denervation occurred during the acute phase of infection for both strains, but persisted chronically only in Be-78 infected animals. Glial cell involvement occurred earlier in animals infected with the Y strain, while animals infected with the Be-78 strain showed reduced glial fibrillary acidic protein immunoreactive area of enteric glial cells in the chronic phase. These results suggest that although both strains cause lesions in the digestive tract, the Y strain is associated with early control of the lesion, while the Be-78 strain results in progressive gut lesions in this model.

  5. The Trypanosoma cruzi nucleolus: a morphometrical analysis of cultured epimastigotes in the exponential and stationary phases.

    Science.gov (United States)

    Nepomuceno-Mejía, Tomás; Lara-Martínez, Reyna; Cevallos, Ana María; López-Villaseñor, Imelda; Jiménez-García, Luis Felipe; Hernández, Roberto

    2010-12-01

    Our group is interested in rRNA and ribosome biogenesis in the parasitic protozoan Trypanosoma cruzi. Epimastigotes represent an extracellular replicative stage of T. cruzi and can be cultured in axenic media. The growth curve of epimastigotes allows assessment of potential differences in the nucleoli of cells undergoing growth-rate transitions. To establish cellular parameters for studying ribosome biogenesis in T. cruzi, a morphometric analysis of the nucleoli of cultured cells in the exponential and stationary phases was conducted. Electron micrograph-based measurements of nuclear sections from independent cells demonstrated that the nucleolar area is over twofold higher in exponentially growing cells, as compared with epimastigotes in the stationary phase. The granular component of the nucleoli of actively growing cells was the main structural element. Cycloheximide moderately reduced the apparent size of the nucleoli without an apparent disruption of their architecture. Our results provide a firm basis for the establishment of an experimental model to study the organization of the nucleolus during the growth and development of T. cruzi. © 2010 Federation of European Microbiological Societies Published by Blackwell Publishing Ltd. All rights reserved.

  6. Evaluation of some organic compounds on bloodstream forms of Trypanosoma cruzi

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    João S. Silva

    1992-09-01

    Full Text Available Accidental transmission of Chagas' disease to man by blood transfusion is a serious problem in Latin-America. This paper describes the testing of several synthetic, semi-synthetic, and natural compounds for their activity against blood trypomastigotes in vitro at 4-C. The compounds embody several types of chemical structures: benzoquinone, naphthoquinone, anthracenequinone, phenanthrenequinone, imidazole, piperazine, quinoline, xanthene, and simple benzenic and naphthalenic derivates. Some of them are for the first time tested against Trypanosoma cruzi. The toxic effect these compounds on this parasite was done by two quite distinct sets of experiments. In one set, the compounds were added to infected blood as ethanolic solution. In this situation the most active one was a furan-1, 2-naphthoquinone, in the same range as gentian violet, a new fact to be considered in the assessment of structure-activity relationships in this class of compounds. In other set, we tentatively evaluated the biological activity of water insoluble compounds by adding them in a pure form without solvent into infected blood. In this way some appear to be very active and it was postulated that the effectiveness of such compounds must result from interactions between them and specific blood components.

  7. Aldo-keto reductase and alcohol dehydrogenase contribute to benznidazole natural resistance in Trypanosoma cruzi.

    Science.gov (United States)

    González, Laura; García-Huertas, Paola; Triana-Chávez, Omar; García, Gabriela Andrea; Murta, Silvane Maria Fonseca; Mejía-Jaramillo, Ana M

    2017-12-01

    The improvement of Chagas disease treatment is focused not only on the development of new drugs but also in understanding mechanisms of action and resistance to drugs conventionally used. Thus, some strategies aim to detect specific changes in proteins between sensitive and resistant parasites and to evaluate the role played in these processes by functional genomics. In this work, we used a natural Trypanosoma cruzi population resistant to benznidazole, which has clones with different susceptibilities to this drug without alterations in the NTR I gene. Using 2DE-gel electrophoresis, the aldo-keto reductase and the alcohol dehydrogenase proteins were found up regulated in the natural resistant clone and therefore their possible role in the resistance to benznidazole and glyoxal was investigated. Both genes were overexpressed in a drug sensitive T. cruzi clone and the biological changes in response to these compounds were evaluated. The results showed that the overexpression of these proteins enhances resistance to benznidazole and glyoxal in T. cruzi. Moreover, a decrease in mitochondrial and cell membrane damage was observed, accompanied by a drop in the intracellular concentration of reactive oxygen species after treatment. Our results suggest that these proteins are involved in the mechanism of action of benznidazole. © 2017 John Wiley & Sons Ltd.

  8. Internalization of components of the host cell plasma membrane during infection by Trypanosoma cruzi

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    Carvalho TMU

    1999-01-01

    Full Text Available Epimastigote and trypomastigote forms of Trypanosoma cruzi attach to the macrophage surface and are internalized with the formation of a membrane bounded vacuole, known as the parasitophorous vacuole (PV. In order to determine if components of the host cell membrane are internalized during formation of the PV we labeled the macrophage surface with fluorescent probes for proteins, lipids and sialic acid residues and then allowed the labeled cells to interact with the parasites. The interaction process was interrupted after 1 hr at 37ºC and the distribution of the probes analyzed by confocal laser scanning microscopy. During attachment of the parasites to the macrophage surface an intense labeling of the attachment regions was observed. Subsequently labeling of the membrane lining the parasitophorous vacuole containing epimastigote and trypomastigote forms was seen. Labeling was not uniform, with regions of intense and light or no labeling. The results obtained show that host cell membrane lipids, proteins and sialoglycoconjugates contribute to the formation of the membrane lining the PV containing epimastigote and trypomastigote T. cruzi forms. Lysosomes of the host cell may participate in the process of PV membrane formation.

  9. Modulation of Trypanosoma cruzi-specific T-cell responses after chemotherapy for chronic Chagas disease

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    María Cecilia Albareda

    2015-05-01

    Full Text Available The aim of this review is to describe the contributions of the knowledge of T-cell responses to the understanding of the physiopathology and the responsiveness to etiological treatment during the chronic phase of Chagas disease. T-helper (Th1 and interleukin (IL-10 Trypanosoma cruzi-specific T-cells have been linked to the asymptomatic phase or to severe clinical forms of the disease, respectively or vice versa, depending on the T. cruzi antigen source, the patient’s location and the performed immunological assays. Parasite-specific T-cell responses are modulated after benznidazole (BZ treatment in chronically T. cruzi-infected subjects in association with a significant decrease in T. cruzi-specific antibodies. Accumulating evidence has indicated that treatment efficacy during experimental infection with T. cruzi results from the combined action of BZ and the activation of appropriate immune responses in the host. However, strong support of this interaction in T. cruzi-infected humans remains lacking. Overall, the quality of T-cell responses might be a key factor in not only disease evolution, but also chemotherapy responsiveness. Immunological parameters are potential indicators of treatment response regardless of achievement of cure. Providing tools to monitor and provide early predictions of treatment success will allow the development of new therapeutic options.

  10. The active transport of histidine and its role in ATP production in Trypanosoma cruzi.

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    Barisón, M J; Damasceno, F S; Mantilla, B S; Silber, A M

    2016-08-01

    Trypanosoma cruzi, the aetiological agent of Chagas's disease, metabolizes glucose, and after its exhaustion, degrades amino acids as energy source. Here, we investigate histidine uptake and its participation in energy metabolism. No putative genes for the histidine biosynthetic pathway have been identified in genome databases of T. cruzi, suggesting that its uptake from extracellular medium is a requirement for the viability of the parasite. From this assumption, we characterized the uptake of histidine in T. cruzi, showing that this amino acid is incorporated through a single and saturable active system. We also show that histidine can be completely oxidised to CO2. This finding, together with the fact that genes encoding the putative enzymes for the histidine - glutamate degradation pathway were annotated, led us to infer its participation in the energy metabolism of the parasite. Here, we show that His is capable of restoring cell viability after long-term starvation. We confirm that as an energy source, His provides electrons to the electron transport chain, maintaining mitochondrial inner membrane potential and O2 consumption in a very efficient manner. Additionally, ATP biosynthesis from oxidative phosphorylation was found when His was the only oxidisable metabolite present, showing that this amino acid is involved in bioenergetics and parasite persistence within its invertebrate host.

  11. Trypanosoma cruzi: partial prevention of the natural infection of guinea pigs with a killed parasite vaccine.

    Science.gov (United States)

    Basombrio, M A

    1990-07-01

    Guinea pigs are natural reservoirs of Chagas' disease. Domestic breeding and local trade of these animals are common practices among andean communities in South America. Infection by Trypanosoma cruzi occurs when the animals live in triatomine-infested houses or yards. The preventive effect of a vaccine consisting of cultured T. cruzi killed by freezing and thawing plus saponin was tested both in mice and in the guinea pig ecosystem. Resistance against T. cruzi challenge in mice was improved by increasing the trypomastigote/epimastigote ratio in live attenuated vaccines but not in killed parasite vaccines. Although the killing of attenuated parasites sharply reduced their immunogenicity for mice, a protective effect against natural T. cruzi infection was detected in guinea pigs. A total of 88 guinea pigs were vaccinated in four intradermal sites on three occasions. Eighty controls received similar inoculations of culture medium plus saponin. All animals were kept in a triatomine-infested yard. Parasitemia was studied with the capillary microhematocrit method. After an exposure time averaging 4 months, natural T. cruzi infection occurred in 55% (44/80) of the controls and in 33% (29/88) of the vaccinated group (P less than 0.01). The number of highly parasitemic guinea pigs was also significantly decreased (6/80 vs 0/88, P less than 0.01). Thus, immunizing protocols which are only partially protective against artificial callenge with T. cruzi may nevertheless constrain the exchange of parasites between natural hosts and vectors.

  12. Using of essential oils in the treatment of mice infected with Trypanosoma evansi

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    Matheus D. Baldissera

    2014-06-01

    Full Text Available Objective. This study aimed to test the effectiveness of copaiba, andiroba and aroeira essential oils for controlling trypanosomosis by Trypanosoma evansi with mice as experimental model. Materials and methods. Sixty-six mice were divided into eleven groups (A to L with six animals each. Group A was the unique composed by healthy and uninfected animals (negative control. Animals in groups B to L were inoculated with 0.1 mL of blood containing 2.7 x 106 trypanosomes. Group B was used as a positive control without treatment. In experiment were tested copaiba (C, D and E, andiroba (F, G and H and aroeira (I, J and L oils at doses of 0.6, 0.8 and 1.0 mL kg-1 to infected mice (T. evansi. Results. These protocols did not provide curative efficacy; however, the mice treated with highest dose of copaiba showed a significant increase in the longevity when compared others groups. Conclusions. Previously in our studies, these essential oils have shown trypanocidal activity in vitro, but when they were tested in vivo in mice infected with T. evansi, this trypanocidal activity, or the curative effect was not found, being only able to prolong the lifespan of the animals treated with copaiba oil.

  13. Trypanosoma equiperdum Low Molecular Weight Proteins As Candidates for Specific Serological Diagnosis of Dourine

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    Mirella Luciani

    2018-03-01

    Full Text Available The diagnosis of dourine can be difficult because the clinical signs of this disease in horses are similar to those of surra, caused by Trypanosoma evansi. Moreover, T. equiperdum and T. evansi are closely related and, so far, they cannot be distinguished using serological tests. In a previous work, the T. equiperdum protein pattern recognized by antibodies from dourine-infected horses and the humoral immune response kinetics were investigated by immunoblotting assay; a total of 20 sera from naturally and experimentally infected horses and from healthy animals were tested. Immunoblotting analysis showed that antibodies from infected horses specifically bind T. equiperdum low molecular weight proteins (from 16 to 35 kDa, which are not recognized by antibodies from uninfected horses. In this work, we tested other 615 sera (7 from naturally infected horses and 608 sera from healthy horses and donkeys: results confirmed the data obtained previously. In addition, six SDS-PAGE bands with molecular weight ranging from 10 to 37 kDa were analyzed by mass spectrometry, in order to identify immunogenic proteins that could be used as biomarkers for the diagnosis of dourine. A total of 167 proteins were identified. Among them, 37 were found unique for T. equiperdum. Twenty-four of them could represent possible candidate diagnostic antigens for the development of serological tests specific for T. equiperdum.

  14. [Seroprevalence of antibodies against Trypanosoma cruzi in 13 departments of Uruguay].

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    Salvatella, R; Calegari, L; Casserone, S; Civila, E; Carbajal, S; Pérez, G; Somma, R; Sampaio, I; Llanes, M E; Conti, M

    1989-08-01

    In 1985 a study was undertaken of the prevalence of Trypanosoma cruzi antibodies in 13 departments of Uruguay where transmission of the parasite by the vector Triatoma infestans persists. A total of 5,924 serum samples were selected using a probabilistic method--3,840 from individuals over the age of 12 (sample I) and 2,084 from subjects who were 12 years old (sample II). The population was classified according to place of residence (capital city, non-capital city, suburban area, and rural area). The percentage of positive sera detected by indirect immunofluorescence in the different departments ranged from 1 to 11%, and overall seroprevalence for the area was 3.4%. Based on the results obtained, it was possible to distinguish three areas: A, with seroprevalence from 6 to 11%; B, 2 to 3.2%, and C, 1 to 1.4%. In sample II from the Departments of Paysandú, Soriano, Flores, Florida, and Durazno, no cases of Chagas' disease were detected, which suggests that there is no active transmission of T. cruzi in this age group in the area studied. The number of persons estimated to have the disease was 36,952, or 1.3% of the total population of Uruguay and 4% of the population in the area surveyed. These seroprevalence figures are similar to those recorded in the province of Entre Ríos, Argentina, and in the neighboring municipalities of Rio Grande do Sul, Brazil.

  15. Notes on the occurrence of Trypanosoma sp. (Kinetoplastida: Trypanosomatidae in freshwater fishes from South Africa

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    Maryke L. Ferreira

    2013-03-01

    Full Text Available A total of 257 fishes from four families, Clariidae, Cichlidae, Cyprinidae and Schilbeidae were collected from three localities: the Sand River Dam, Swaziland; the Nylsvlei Nature Reserve, South Africa and the Vaal Dam and Vaal River Barrage, South Africa. Only fishes (n= 154 from Clariidae and Cichlidae were found to be infected with trypanosomes. A total of 221 Clarias gariepinus (Burchell 1822 were collected from the Vaal Dam and Vaal Barrage area, South Africa. Of these, 74%(89/121 were infected with trypanosomes from the Vaal Dam and 63%(63/100 from the Vaal River Barrage, with no seasonal infection pattern. A prevalence of 25%(1/4 was found in C. gariepinus from the Sand River Dam, Swaziland, and a 50% (1/2 prevalence was found in Tilapia sparrmanii from the Nylsvlei Nature Reserve, South Africa. Standard measurements conformed closely to the morphometric and morphological descriptions of Trypanosoma mukasai. This article provides new locality records for T. mukasai from the Vaal Dam, Vaal River Barrage and Nylsvlei Nature Reserve (South Africa and the Sand River Dam (Swaziland. Tilapia sparrmanii collected in the Sand River Dam in Swaziland is also noted as a new host record.

  16. Astrocyte Apoptosis and HIV Replication Are Modulated in Host Cells Coinfected with Trypanosoma cruzi

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    Javier M. Urquiza

    2017-08-01

    Full Text Available The protozoan Trypanosoma cruzi is the etiological agent of Chagas disease. In immunosuppressed individuals, as it occurs in the coinfection with human immunodeficiency virus (HIV, the central nervous system may be affected. In this regard, reactivation of Chagas disease is severe and often lethal, and it accounts for meningoencephalitis. Astrocytes play a crucial role in the environment maintenance of healthy neurons; however, they can host HIV and T. cruzi. In this report, human astrocytes were infected in vitro with both genetically modified-pathogens to express alternative fluorophore. As evidenced by fluorescence microscopy and flow cytometry, HIV and T. cruzi coexist in the same astrocyte, likely favoring reciprocal interactions. In this context, lower rates of cell death were observed in both T. cruzi monoinfected-astrocytes and HIV-T. cruzi coinfection in comparison with those infected only with HIV. The level of HIV replication is significantly diminished under T. cruzi coinfection, but without affecting the infectivity of the HIV progeny. This interference with viral replication appears to be related to the T. cruzi multiplication rate or its increased intracellular presence but does not require their intracellular cohabitation or infected cell-to-cell contact. Among several Th1/Th2/Th17 profile-related cytokines, only IL-6 was overexpressed in HIV-T. cruzi coinfection exhibiting its cytoprotective role. This study demonstrates that T. cruzi and HIV are able to coinfect astrocytes thus altering viral replication and apoptosis.

  17. Immunoglobulin M antibodies against CRA and FRA recombinant antigens of Trypanosoma cruzi in chronic chagasic patients.

    Science.gov (United States)

    Vasconcelos, Romero H T; Azevedo, Elisa A N; Cavalcanti, Maria G A M; Silva, Edimilson D; Ferreira, Antonio G P; Morais, Clarice N L; Gomes, Yara M

    2011-05-01

    Previous works of our research group have demonstrated aspects of the humoral immune response of chronic Chagas disease using the cytoplasmatic repetitive antigen (CRA) and the flagellar repetitive antigen (FRA) of Trypanosoma cruzi. The aim of this work was to analyze the presence of specific immunoglobulin M (IgM) antibodies in chronic chagasic patients using these recombinant antigens of T. cruzi. The positivity of IgM in chronic chagasic patients against CRA and FRA antigens was determined by indirect enzyme-linked immunosorbent assay. We reported no statistical significant differences between the levels of IgM for both recombinant antigens and the different chronic clinical forms of Chagas disease. However, a small proportion of chronic chagasic patients analyzed in this study was positive for this antibody isotype. The findings of this study indicate that the IgM antibodies cannot be used to elucidate the differences in the profile of humoral immune response among chronic chagasic patients with different clinical forms using the CRA and FRA recombinant antigens of T. cruzi. Copyright © 2011 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.

  18. Blocking variant surface glycoprotein synthesis alters endoplasmic reticulum exit sites/Golgi homeostasis in Trypanosoma brucei.

    Science.gov (United States)

    Ooi, Cher-Pheng; Smith, Terry K; Gluenz, Eva; Wand, Nadina Vasileva; Vaughan, Sue; Rudenko, Gloria

    2018-06-01

    The predominant secretory cargo of bloodstream form Trypanosoma brucei is variant surface glycoprotein (VSG), comprising ~10% total protein and forming a dense protective layer. Blocking VSG translation using Morpholino oligonucleotides triggered a precise pre-cytokinesis arrest. We investigated the effect of blocking VSG synthesis on the secretory pathway. The number of Golgi decreased, particularly in post-mitotic cells, from 3.5 ± 0.6 to 2.0 ± 0.04 per cell. Similarly, the number of endoplasmic reticulum exit sites (ERES) in post-mitotic cells dropped from 3.9 ± 0.6 to 2.7 ± 0.1 eight hours after blocking VSG synthesis. The secretory pathway was still functional in these stalled cells, as monitored using Cathepsin L. Rates of phospholipid and glycosylphosphatidylinositol-anchor biosynthesis remained relatively unaffected, except for the level of sphingomyelin which increased. However, both endoplasmic reticulum and Golgi morphology became distorted, with the Golgi cisternae becoming significantly dilated, particularly at the trans-face. Membrane accumulation in these structures is possibly caused by reduced budding of nascent vesicles due to the drastic reduction in the total amount of secretory cargo, that is, VSG. These data argue that the total flux of secretory cargo impacts upon the biogenesis and maintenance of secretory structures and organelles in T. brucei, including the ERES and Golgi. © 2018 The Authors. Traffic published by John Wiley & Sons Ltd.

  19. A novel ABCG-like transporter of Trypanosoma cruzi is involved in natural resistance to benznidazole

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    Bianca Zingales

    2015-05-01

    Full Text Available Benznidazole (BZ is one of the two drugs used for Chagas disease treatment. Nevertheless therapeutic failures of BZ have been reported, which were mostly attributed to variable drug susceptibility among Trypanosoma cruzi strains. ATP-binding cassette (ABC transporters are involved in a variety of translocation processes and some members have been implicated in drug resistance. Here we report the characterisation of the first T. cruzi ABCG transporter gene, named TcABCG1, which is over-expressed in parasite strains naturally resistant to BZ. Comparison of TcABCG1 gene sequence of two TcI BZ-resistant strains with CL Brener BZ-susceptible strain showed several single nucleotide polymorphisms, which determined 11 amino acid changes. CL Brener transfected with TcI transporter genes showed 40-47% increased resistance to BZ, whereas no statistical significant increment in drug resistance was observed when CL Brener was transfected with the homologous gene. Only in the parasites transfected with TcI genes there was 2-2.6-fold increased abundance of TcABCG1 transporter protein. The analysis in wild type strains also suggests that the level of TcABCG1 transporter is related to BZ natural resistance. The characteristics of untranslated regions of TcABCG1 genes of BZ-susceptible and resistant strains were investigated by computational tools.

  20. Relationship between Trypanosoma brucei rhodesiense genetic diversity and clinical spectrum among sleeping sickness patients in Uganda.

    Science.gov (United States)

    Kato, Charles D; Mugasa, Claire M; Nanteza, Ann; Matovu, Enock; Alibu, Vincent P

    2017-10-27

    Human African trypanosomiasis (HAT) due to Trypanosoma brucei rhodesiense in East and southern Africa is reported to be clinically diverse. We tested the hypothesis that this clinical diversity is associated with a variation in trypanosome genotypes. Trypanosome DNA isolated from HAT patients was genotyped using 7 microsatellite markers directly from blood spotted FTA cards following a whole genome amplification. All markers were polymorphic and identified 17 multi-locus genotypes with 56% of the isolates having replicate genotypes. We did not observe any significant clustering between isolates and bootstrap values across major tree nodes were insignificant. When genotypes were compared among patients with varying clinical presentation or outcome, replicate genotypes were observed at both extremes showing no significant association between genetic diversity and clinical outcome. Our study shows that T. b. rhodesiense isolates are homogeneous within a focus and that observed clinical diversity may not be associated with parasite genetic diversity. Other factors like host genetics and environmental factors might be involved in determining clinical diversity. Our study may be important in designing appropriate control measures that target the parasite.