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An immunoproteomics approach to identify Leishmania infantum proteins to be applied for the diagnosis of visceral leishmaniasis and human immunodeficiency virus co-infection
- Amanda S. Machado, Fernanda F. Ramos, João A. Oliveira-da-Silva, Thaís T. O. Santos, Grasiele S. V. Tavares, Lourena E. Costa, Daniela P. Lage, André Teixeira-Ferreira, Jonas Perales, Ana Paula Fernandes, Ricardo L. F. Moreira, Mariana C. Duarte, Unaí Tupinambás, Rachel B. Caligiorne, Gláucia F. Cota, Eduardo A. F. Coelho, Fernanda Ludolf
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- Journal:
- Parasitology / Volume 147 / Issue 9 / August 2020
- Published online by Cambridge University Press:
- 20 April 2020, pp. 932-939
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The co-infection between visceral leishmaniasis (VL) and human immunodeficiency virus (HIV) has increased in several countries in the world. The current serological tests are not suitable since they present low sensitivity to detect the most of VL/HIV cases, and a more precise diagnosis should be performed. In this context, in the present study, an immunoproteomics approach was performed using Leishmania infantum antigenic extracts and VL, HIV and VL/HIV patients sera, besides healthy subjects samples; aiming to identify antigenic markers for these clinical conditions. Results showed that 43 spots were recognized by antibodies in VL and VL/HIV sera, and 26 proteins were identified by mass spectrometry. Between them, β-tubulin was expressed, purified and tested in ELISA experiments as a proof of concept for validation of our immunoproteomics findings and results showed high sensitivity and specificity values to detect VL and VL/HIV patients. In conclusion, the identified proteins in the present work could be considered as candidates for future studies aiming to improvement of the diagnosis of VL and VL/HIV co-infection.
Screening diagnostic candidates from Leishmania infantum proteins for human visceral leishmaniasis using an immunoproteomics approach
- Daniela P. Lage, Fernanda Ludolf, Patrícia C. Silveira, Amanda S. Machado, Fernanda F. Ramos, Daniel S. Dias, Patrícia A. F. Ribeiro, Lourena E. Costa, Danniele L. Vale, Grasiele S. V. Tavares, Vívian T. Martins, Miguel A. Chávez-Fumagalli, Rachel B. Caligiorne, Ana T. Chaves, Denise U. Gonçalves, Manoel O. C. Rocha, Mariana C. Duarte, Eduardo A. F. Coelho
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- Journal:
- Parasitology / Volume 146 / Issue 11 / September 2019
- Published online by Cambridge University Press:
- 13 June 2019, pp. 1467-1476
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There is no suitable vaccine against human visceral leishmaniasis (VL) and available drugs are toxic and/or present high cost. In this context, diagnostic tools should be improved for clinical management and epidemiological evaluation of disease. However, the variable sensitivity and/or specificity of the used antigens are limitations, showing the necessity to identify new molecules to be tested in a more sensitive and specific serology. In the present study, an immunoproteomics approach was performed in Leishmania infantum promastigotes and amastigotes employing sera samples from VL patients. Aiming to avoid undesired cross-reactivity in the serological assays, sera from Chagas disease patients and healthy subjects living in the endemic region of disease were also used in immunoblottings. The most reactive spots for VL samples were selected, and 29 and 21 proteins were identified in the promastigote and amastigote extracts, respectively. Two of them, endonuclease III and GTP-binding protein, were cloned, expressed, purified and tested in ELISA experiments against a large serological panel, and results showed high sensitivity and specificity values for the diagnosis of disease. In conclusion, the identified proteins could be considered in future studies as candidate antigens for the serodiagnosis of human VL.
High-through identification of T cell-specific phage-exposed mimotopes using PBMCs from tegumentary leishmaniasis patients and their use as vaccine candidates against Leishmania amazonensis infection
- Gerusa B. Carvalho, Lourena E. Costa, Daniela P. Lage, Fernanda F. Ramos, Thaís T. O. Santos, Patrícia A. F. Ribeiro, Daniel S. Dias, Beatriz C. S. Salles, Mariana P. Lima, Lívia M. Carvalho, Ana C. S. Dias, Patrícia T. Alves, Michelle L. Franklin, Renata A. M. Silva, Mariana C. Duarte, Daniel Menezes-Souza, Bruno M. Roatt, Miguel A. Chávez-Fumagalli, Luiz Ricardo Goulart, Antonio L. Teixeira, Eduardo A. F. Coelho
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- Journal:
- Parasitology / Volume 146 / Issue 3 / March 2019
- Published online by Cambridge University Press:
- 10 September 2018, pp. 322-332
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In the current study, phage-exposed mimotopes as targets against tegumentary leishmaniasis (TL) were selected by means of bio-panning cycles employing sera of TL patients and healthy subjects, besides the immune stimulation of peripheral blood mononuclear cells (PBMCs) collected from untreated and treated TL patients and healthy subjects. The clones were evaluated regarding their specific interferon-γ (IFN-γ) and interleukin-4 (IL-4) production in the in vitro cultures, and selectivity and specificity values were calculated, and those presenting the best results were selected for the in vivo experiments. Two clones, namely A4 and A8, were identified and used in immunization protocols from BALB/c mice to protect against Leishmania amazonensis infection. Results showed a polarized Th1 response generated after vaccination, being based on significantly higher levels of IFN-γ, IL-2, IL-12, tumour necrosis factor-α (TNF-α) and granulocyte-macrophage colony-stimulating factor (GM-CSF); which were associated with lower production of specific IL-4, IL-10 and immunoglobulin G1 (IgG1) antibodies. Vaccinated mice presented significant reductions in the parasite load in the infected tissue and distinct organs, when compared with controls. In conclusion, we presented a strategy to identify new mimotopes able to induce Th1 response in PBMCs from TL patients and healthy subjects, and that were successfully used to protect against L. amazonensis infection.
Zygosity Differences in Height and Body Mass Index of Twins From Infancy to Old Age: A Study of the CODATwins Project
- Aline Jelenkovic, Yoshie Yokoyama, Reijo Sund, Chika Honda, Leonie H Bogl, Sari Aaltonen, Fuling Ji, Feng Ning, Zengchang Pang, Juan R. Ordoñana, Juan F. Sánchez-Romera, Lucia Colodro-Conde, S. Alexandra Burt, Kelly L. Klump, Sarah E. Medland, Grant W. Montgomery, Christian Kandler, Tom A. McAdams, Thalia C. Eley, Alice M. Gregory, Kimberly J. Saudino, Lise Dubois, Michel Boivin, Adam D. Tarnoki, David L. Tarnoki, Claire M. A. Haworth, Robert Plomin, Sevgi Y. Öncel, Fazil Aliev, Maria A. Stazi, Corrado Fagnani, Cristina D’Ippolito, Jeffrey M. Craig, Richard Saffery, Sisira H. Siribaddana, Matthew Hotopf, Athula Sumathipala, Fruhling Rijsdijk, Timothy Spector, Massimo Mangino, Genevieve Lachance, Margaret Gatz, David A. Butler, Gombojav Bayasgalan, Danshiitsoodol Narandalai, Duarte L Freitas, José Antonio Maia, K. Paige Harden, Elliot M. Tucker-Drob, Bia Kim, Youngsook Chong, Changhee Hong, Hyun Jung Shin, Kaare Christensen, Axel Skytthe, Kirsten O. Kyvik, Catherine A. Derom, Robert F. Vlietinck, Ruth J. F. Loos, Wendy Cozen, Amie E. Hwang, Thomas M. Mack, Mingguang He, Xiaohu Ding, Billy Chang, Judy L. Silberg, Lindon J. Eaves, Hermine H. Maes, Tessa L. Cutler, John L. Hopper, Kelly Aujard, Patrik K. E. Magnusson, Nancy L. Pedersen, Anna K. Dahl Aslan, Yun-Mi Song, Sarah Yang, Kayoung Lee, Laura A. Baker, Catherine Tuvblad, Morten Bjerregaard-Andersen, Henning Beck-Nielsen, Morten Sodemann, Kauko Heikkilä, Qihua Tan, Dongfeng Zhang, Gary E. Swan, Ruth Krasnow, Kerry L. Jang, Ariel Knafo-Noam, David Mankuta, Lior Abramson, Paul Lichtenstein, Robert F. Krueger, Matt McGue, Shandell Pahlen, Per Tynelius, Glen E. Duncan, Dedra Buchwald, Robin P. Corley, Brooke M. Huibregtse, Tracy L. Nelson, Keith E. Whitfield, Carol E. Franz, William S. Kremen, Michael J. Lyons, Syuichi Ooki, Ingunn Brandt, Thomas Sevenius Nilsen, Fujio Inui, Mikio Watanabe, Meike Bartels, Toos C. E. M. van Beijsterveldt, Jane Wardle, Clare H. Llewellyn, Abigail Fisher, Esther Rebato, Nicholas G. Martin, Yoshinori Iwatani, Kazuo Hayakawa, Joohon Sung, Jennifer R. Harris, Gonneke Willemsen, Andreas Busjahn, Jack H. Goldberg, Finn Rasmussen, Yoon-Mi Hur, Dorret I. Boomsma, Thorkild I. A. Sørensen, Jaakko Kaprio, Karri Silventoinen
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- Journal:
- Twin Research and Human Genetics / Volume 18 / Issue 5 / October 2015
- Published online by Cambridge University Press:
- 04 September 2015, pp. 557-570
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A trend toward greater body size in dizygotic (DZ) than in monozygotic (MZ) twins has been suggested by some but not all studies, and this difference may also vary by age. We analyzed zygosity differences in mean values and variances of height and body mass index (BMI) among male and female twins from infancy to old age. Data were derived from an international database of 54 twin cohorts participating in the COllaborative project of Development of Anthropometrical measures in Twins (CODATwins), and included 842,951 height and BMI measurements from twins aged 1 to 102 years. The results showed that DZ twins were consistently taller than MZ twins, with differences of up to 2.0 cm in childhood and adolescence and up to 0.9 cm in adulthood. Similarly, a greater mean BMI of up to 0.3 kg/m2 in childhood and adolescence and up to 0.2 kg/m2 in adulthood was observed in DZ twins, although the pattern was less consistent. DZ twins presented up to 1.7% greater height and 1.9% greater BMI than MZ twins; these percentage differences were largest in middle and late childhood and decreased with age in both sexes. The variance of height was similar in MZ and DZ twins at most ages. In contrast, the variance of BMI was significantly higher in DZ than in MZ twins, particularly in childhood. In conclusion, DZ twins were generally taller and had greater BMI than MZ twins, but the differences decreased with age in both sexes.
The CODATwins Project: The Cohort Description of Collaborative Project of Development of Anthropometrical Measures in Twins to Study Macro-Environmental Variation in Genetic and Environmental Effects on Anthropometric Traits
- Karri Silventoinen, Aline Jelenkovic, Reijo Sund, Chika Honda, Sari Aaltonen, Yoshie Yokoyama, Adam D. Tarnoki, David L. Tarnoki, Feng Ning, Fuling Ji, Zengchang Pang, Juan R. Ordoñana, Juan F. Sánchez-Romera, Lucia Colodro-Conde, S. Alexandra Burt, Kelly L. Klump, Sarah E. Medland, Grant W. Montgomery, Christian Kandler, Tom A. McAdams, Thalia C. Eley, Alice M. Gregory, Kimberly J. Saudino, Lise Dubois, Michel Boivin, Claire M. A. Haworth, Robert Plomin, Sevgi Y. Öncel, Fazil Aliev, Maria A. Stazi, Corrado Fagnani, Cristina D’Ippolito, Jeffrey M. Craig, Richard Saffery, Sisira H. Siribaddana, Matthew Hotopf, Athula Sumathipala, Timothy Spector, Massimo Mangino, Genevieve Lachance, Margaret Gatz, David A. Butler, Gombojav Bayasgalan, Danshiitsoodol Narandalai, Duarte L. Freitas, José Antonio Maia, K. Paige Harden, Elliot M. Tucker-Drob, Kaare Christensen, Axel Skytthe, Kirsten O. Kyvik, Changhee Hong, Youngsook Chong, Catherine A. Derom, Robert F. Vlietinck, Ruth J. F. Loos, Wendy Cozen, Amie E. Hwang, Thomas M. Mack, Mingguang He, Xiaohu Ding, Billy Chang, Judy L. Silberg, Lindon J. Eaves, Hermine H. Maes, Tessa L. Cutler, John L. Hopper, Kelly Aujard, Patrik K. E. Magnusson, Nancy L. Pedersen, Anna K. Dahl Aslan, Yun-Mi Song, Sarah Yang, Kayoung Lee, Laura A. Baker, Catherine Tuvblad, Morten Bjerregaard-Andersen, Henning Beck-Nielsen, Morten Sodemann, Kauko Heikkilä, Qihua Tan, Dongfeng Zhang, Gary E. Swan, Ruth Krasnow, Kerry L. Jang, Ariel Knafo-Noam, David Mankuta, Lior Abramson, Paul Lichtenstein, Robert F. Krueger, Matt McGue, Shandell Pahlen, Per Tynelius, Glen E. Duncan, Dedra Buchwald, Robin P. Corley, Brooke M. Huibregtse, Tracy L. Nelson, Keith E. Whitfield, Carol E. Franz, William S. Kremen, Michael J. Lyons, Syuichi Ooki, Ingunn Brandt, Thomas Sevenius Nilsen, Fujio Inui, Mikio Watanabe, Meike Bartels, Toos C. E. M. van Beijsterveldt, Jane Wardle, Clare H. Llewellyn, Abigail Fisher, Esther Rebato, Nicholas G. Martin, Yoshinori Iwatani, Kazuo Hayakawa, Finn Rasmussen, Joohon Sung, Jennifer R. Harris, Gonneke Willemsen, Andreas Busjahn, Jack H. Goldberg, Dorret I. Boomsma, Yoon-Mi Hur, Thorkild I. A. Sørensen, Jaakko Kaprio
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- Twin Research and Human Genetics / Volume 18 / Issue 4 / August 2015
- Published online by Cambridge University Press:
- 27 May 2015, pp. 348-360
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For over 100 years, the genetics of human anthropometric traits has attracted scientific interest. In particular, height and body mass index (BMI, calculated as kg/m2) have been under intensive genetic research. However, it is still largely unknown whether and how heritability estimates vary between human populations. Opportunities to address this question have increased recently because of the establishment of many new twin cohorts and the increasing accumulation of data in established twin cohorts. We started a new research project to analyze systematically (1) the variation of heritability estimates of height, BMI and their trajectories over the life course between birth cohorts, ethnicities and countries, and (2) to study the effects of birth-related factors, education and smoking on these anthropometric traits and whether these effects vary between twin cohorts. We identified 67 twin projects, including both monozygotic (MZ) and dizygotic (DZ) twins, using various sources. We asked for individual level data on height and weight including repeated measurements, birth related traits, background variables, education and smoking. By the end of 2014, 48 projects participated. Together, we have 893,458 height and weight measures (52% females) from 434,723 twin individuals, including 201,192 complete twin pairs (40% monozygotic, 40% same-sex dizygotic and 20% opposite-sex dizygotic) representing 22 countries. This project demonstrates that large-scale international twin studies are feasible and can promote the use of existing data for novel research purposes.
Acute effects on the thyroid gland after non-directed radiation therapy in children and adolescents
- C. C. Bonato, H. B. Dias, M. da S. Alves, L. O. Duarte, T. M. Dias, M. O. Dalenogare, C. C. B. Viegas, R. H. Elnecave
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- Journal:
- Journal of Radiotherapy in Practice / Volume 13 / Issue 3 / September 2014
- Published online by Cambridge University Press:
- 12 June 2013, pp. 310-316
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Background
During radiation therapy, unwanted scatter to healthy tissues outside the target field may occur. Children and adolescents are more sensitive to radiation injury, and the thyroid gland is particularly susceptible to these effects.
PurposeTo assess acute changes in thyroid function and volume in children and adolescents undergoing radiotherapy for a variety of non-thyroid cancers.
Materials and MethodsThirty-one children and adolescents underwent radiation therapy of various body areas in which the thyroid was not included. Thyroid-stimulating hormone (TSH), thyroxine (T4), free thyroxine (fT4), triiodothyronine (T3), anti-thyroperoxidase antibodies and thyroglobulin were measured before, on the last day and at 1 and 3 months after the end of radiotherapy. Ultrasound scans were taken and 6- and 24-hour 131I uptake was measured before and after treatment. The scattered dose to the thyroid region was estimated with a treatment planning system or measured with thermoluminescent dosimeters.
ResultsThe median radiation dose scattered to the thyroid was 296·6 cGy (IQR 16·7–1,709·0). Levels of TSH (p = 0·575), T4 (p = 0·950), fT4 (p = 0·510), T3 (p = 0·842), thyroglobulin (p = 0·620) and anti-thyroid peroxidase antibodies (p = 0·546) were statistically similar at all four time points. There were no differences between pre- and post-radiotherapy thyroid volume and 131I uptake (p = 0·692 and 0·92, respectively).
ConclusionMore sensitive methods may be required to ascertain whether acute injury to the follicular epithelium occurs with lower radiation doses scattered to the thyroid.
Contributors
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- By Jennifer Alvarez, Ananda B. Amstadter, Metin Başoğlu, David M. Benedek, Charles C. Benight, George A. Bonanno, Evelyn J. Bromet, Richard A. Bryant, Barbara Lopes Cardozo, M. L. Somchai Chakkraband, Claude Chemtob, Roman Cieslak, Lauren M. Conoscenti, Joan M. Cook, Judith Cukor, Carla Kmett Danielson, JoAnn Difede, Charles DiMaggio, Anja J.E. Dirkzwager, Cristiane S. Duarte, Jon D. Elhai, Diane L. Elmore, Yael L.E. Errera, Julian D. Ford, Carol S. Fullerton, Sandro Galea, Freya Goodhew, Neil Greenberg, Lindsay Greene, Linda Grievink, Michael J. Gruber, Sumati Gupta, Johan M. Havenaar, Alesia O. Hawkins, Clare Henn-Haase, Kimberly Eaton Hoagwood, Christina W. Hoven, Sabra S. Inslicht, Krzysztof Kaniasty, Ronald C. Kessler, Rachel Kimerling, Richard V. King, Rolf J. Kleber, Jessica Mass Levitt, Brett T. Litz, Maria Livanou, Katelyn P. Mack, Paula Madrid, Shira Maguen, Paul Maguire, Donald J. Mandell, Charles R. Marmar, Andrea R. Maxwell, Shannon E. McCaslin, Alexander C. McFarlane, Thomas J. Metzler, Summer Nelson, Yuval Neria, Elana Newman, Thomas C. Neylan, Fran H. Norris, Carol S. North, Lawrence A. Palinkas, Benjaporn Panyayong, Maria Petukhova, Betty Pfefferbaum, Marleen Radigan, Beverley Raphael, James Rodriguez, G. James Rubin, Kenneth J. Ruggiero, Ebru Şalcıoğlu, Nancy A. Sampson, Arieh Y. Shalev, Bruce Shapiro, Laura M. Stough, Prawate Tantipiwatanaskul, Warunee Thienkrua, Phebe Tucker, J. Blake Turner, Robert J. Ursano, Bellis van den Berg, Peter G. van der Velden, Frits van Griensven, Miranda Van Hooff, Edward Waldrep, Philip S. Wang, Simon Wessely, Leslie H. Wind, C. Joris Yzermans, Heidi M. Zinzow
- Edited by Yuval Neria, Columbia University, New York, Sandro Galea, University of Michigan, Ann Arbor, Fran H. Norris
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- Book:
- Mental Health and Disasters
- Published online:
- 07 May 2010
- Print publication:
- 20 July 2009, pp xi-xvi
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