159 results on '"Ricardo Toshio Fujiwara"'
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2. Leishmaniose Visceral Canina: um problema de saúde pública em expansão
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Thiago Pasqua Narciso, Ana Paula Peconick, Angélica Terezinha Barth Wouters, Tarcísio de Freitas Milagres, Ricardo Toshio Fujiwara, Luiza Almeida Figueiredo, Agostinho Gonçalves Viana, Ingrid Marciano Alvarenga, Richardson Costa Carvalho, Thales Augusto Barçante, José Cherem, and Joziana Muniz de Paiva Barçante
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General Medicine - Abstract
As leishmanioses são doenças negligenciadas de grande importância em saúde pública. Esta doença, causada por parasitos do gênero Leishmania e transmitidos por insetos vetores pode se manifestar sob as formas visceral, cutânea e muco-cutânea. A leishmaniose visceral ganhou importante destaque na abordagem da “Saúde Única” em função da interface humana, veterinária e ambiental. Neste contexto, a detecção de novas áreas com ocorrência de casos autóctones de LV canina é o marco para iniciar a investigação, vigilância e monitoramento epidemiológico. O município de Lavras era considerado área silenciosa e não vulnerável para leishmaniose visceral canina até o ano de 2013. Diante da necessidade de confirmação da infecção canina, o objetivo do presente trabalho foi confirmar a ocorrência de casos autóctones de LV canina no município de Lavras – MG, com técnicas parasitológicas, sorológicas e moleculares. A partir da Reação em Cadeia da Polimerase em tempo real – qPCR, foi analisada a carga parasitária de amostras de baço e medula óssea de nove cães naturalmente infectados, sabidamente positivos para LVC nos testes DPP® e EIE LVC Bio-Manguinhos®. Foram observadas quatro amostras positivas para Leishmania infantum em medula óssea, e cinco amostras positivas de baço. Não houve diferença significativa para os valores de carga parasitária entre os dois tipos de tecido analisados. A confirmação da infecção por L. infantum em cães realizada neste estudo é a primeira na macrorregião de Lavras - sul do estado de Minas Gerais, e serve de alerta para implementação das ações de vigilância e controle, no sentido de evitar a dispersão da doença e a ocorrência de casos humanos.
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- 2023
3. Chemokines and chemokine receptors: Insights from human disease and experimental models of helminthiasis
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Lucas, Kraemer, Derek M, McKay, Remo Castro, Russo, and Ricardo Toshio, Fujiwara
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Helminths ,Endocrinology, Diabetes and Metabolism ,Immunology ,Helminthiasis ,Animals ,Humans ,Immunology and Allergy ,Receptors, Chemokine ,Chemokines ,Models, Theoretical ,General Biochemistry, Genetics and Molecular Biology ,Host-Parasite Interactions - Abstract
Infection with helminth parasites affects more than 1.5 billion people and is concentrated in global areas of extreme poverty, having a significant impact on public health, social life and the economy. Upon entry into the host, helminth parasites often migrate through specific tissues triggering host immunity. The immune response triggered by helminth infections is complex and depends on parasite load, site of infection, acuteness/chronicity of the infection and is species-dependent. In general, susceptibility or resistance to the infection involves the participation of the innate immune response and then the balance between several effector CD4
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- 2022
4. Vaccine Development for Human Leishmaniasis
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Marianna de Carvalho Clímaco, Lucas Kraemer, and Ricardo Toshio Fujiwara
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The development of vaccines for human leishmaniasis is one of the most important approaches for effectively controlling and/or eradicating the several forms of the disease. Based on the knowledge obtained from the practice of leishmanization and its protective immune response, several strategies have been used to develop vaccines against Leishmania species, such as the use of whole killed and attenuated parasites, recombinant proteins, and DNA vaccines. An ideal vaccine should be safe, effective, and immunogenic. Although several candidates have achieved safety and some level of effectiveness, the current challenge in the development of prophylactic vaccines is to achieve long-lasting immune protection by generating a robust and irreversible Th1 adaptive immune response in the host, with rapid recruitment of memory and effectors T cells at key acute points of infection. However, despite all efforts over the years, due to the antigenic diversity of the parasite and the complexity of the host’s immune response, human vaccine trials have been disappointing in mediating long-term immunity against sandfly-delivered infection. Therefore, more investments in this field should be carried out to translate preclinical findings from mice to humans through effective vaccine development strategies.
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- 2023
5. COVID-19 Pandemic Impacted the Actions of the Schistosomiasis Control Program in Brazil: Is the Goal of Controlling the Disease by 2030 at Risk?
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Lucas Almeida Andrade, Wandklebson Silva da Paz, Rosália E. Santos Ramos, Welde N. Borges de Santana, Thuelly Juvêncio da Rocha, Flávia Silva Damasceno, Allan Dantas dos Santos, Débora dos Santos Tavares, Rodrigo Feliciano do Carmo, Carlos Dornels Freire de Souza, Deborah Aparecida Negrão-Corrêa, Ricardo Toshio Fujiwara, Abelardo Silva-Júnior, Wagnner José Nascimento Porto, and Márcio Bezerra-Santos
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- 2023
6. Tissue eosinophilia correlates with mice susceptibility, granuloma formation and damage during Toxocara canis infection
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Thaís Leal-Silva, Camila de Almeida Lopes, Flaviane Vieira-Santos, Fabrício Marcus Silva Oliveira, Lucas Kraemer, Luiza de Lima Silva Padrão, Chiara Cássia Oliveira Amorim, Jorge Lucas Nascimento Souza, Fernando Sérgio Barbosa, Milene Alvarenga Rachid, Remo Castro Russo, Ricardo Toshio Fujiwara, and Lilian Lacerda Bueno
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Infectious Diseases ,Animal Science and Zoology ,Parasitology - Abstract
An increase in peripheral blood eosinophils in helminth infections is expected, and these cells are known to promote immunity against these parasites. However, studies have suggested that in some specific helminths, eosinophils may promote the needs and longevity of these parasites, and their role in these infections remains undefined, including in Toxocara canis infection. Thus, this study aimed to investigate the role of eosinophils in the context of larval migration of T. canis and the immunopathological aspects of infection. For this, we used wild-type mice and mice genetically deficient for the transcription factor GATA-binding factor 1 (GATA1−/−), infected with 1000 eggs of T. canis. At 0, 3, 14 and 63 days post-infection, parasite load, tissue cytokine production, leucocyte profile, bronchoalveolar lavage cells and histopathological analyses were carried out. Collectively, our results demonstrate that the presence of eosinophils mediates susceptibility to T. canis, inducing leucocytosis and the formation of granulomas, increasing the pulmonary and cerebral parasite load, and reducing the number of neutrophils, which may be necessary to control the infection.
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- 2022
7. Biological activity of 1,2,3-triazole-2-amino-1,4-naphthoquinone derivatives and their evaluation as therapeutic strategy for malaria control
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Renata Maria Costa Souza, Lilian Maria Lapa Montenegro Pimentel, Laryssa Kathleen Mendonça Ferreira, Valéria Rêgo Alves Pereira, Aline Caroline Da Silva Santos, Willyenne Marília Dantas, Carla Jasmine Oliveira Silva, Ramayana Morais De Medeiros Brito, José Lucas Andrade, Valter Ferreira De Andrade-Neto, Ricardo Toshio Fujiwara, Lilian Lacerda Bueno, Valdemiro Amaro Silva Junior, Lindomar Pena, Celso Amorim Camara, Brijesh Rathi, and Ronaldo Nascimento De Oliveira
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Pharmacology ,Organic Chemistry ,Drug Discovery ,General Medicine - Published
- 2023
8. Cytokine response to resistance training sessions performed after different recovery intervals
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Ricardo Toshio Fujiwara, Fernando Vitor Lima, Nathalia Maria Resende, Luciana Maria Oliveira, Bruno Pena Couto, Marcos Daniel Motta Drummond, and Karine Naves de Oliveira Goulart
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medicine.medical_specialty ,Sports medicine ,business.industry ,Resistance training ,Repetition maximum ,Repeated measures design ,Squat ,Recovery interval ,Cytokine response ,Anesthesia ,Medicine ,Orthopedics and Sports Medicine ,business ,Leg press - Abstract
This study aimed to compare the inflammatory response during resistance training (RT) sessions performed after recovery intervals of 24 h, 48 h, and 72 h. Thirteen resistance-trained male individuals (24 ± 2 years) performed three experimental conditions, each one including two sessions with a recovery interval of 24 h, 48 h or 72 h, in a randomized order. The RT sessions consisted of five sets of 8–10 repetition maximum (RM) on squat and leg press exercises. The resistance was equal in the two sessions of each condition and repetitions were performed until concentric failure. Volume load (repetitions × resistance lifted) were compared between sessions. Blood was collected pre and immediately post each RT session, for analyses of IL-6, IL-10, IL-12/IL-23p40, IL-17, TNF-α, and IFN-y. A (2 × 3) repeated measures ANOVA compared the volume load and ratio (immediately post/pre) values of all cytokines within time and conditions (α = 0.05). A significant interaction was observed for volume load, which was reduced at post-recovery interval compared to pre for 24 h condition (p = 0.014), although no differences were observed for 48 h (p = 0.231) and 72 h (p = 0.237) conditions. No significant interaction was evident for any cytokine (p > 0.05), though a significant time main effect was observed for IL-6, which was significantly higher at post-recovery interval compared to pre for all conditions. Despite reduced volume load when a resistance protocol to failure is performed at 24 h, no significant differences were observed on the inflammatory responses. Thus, a similar inflammatory profile is evident in trained men regardless recovery interval between resistance training sessions.
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- 2021
9. Accessing the Variability of Multicopy Genes in Complex Genomes using Unassembled Next-Generation Sequencing Reads: The Case of Trypanosoma cruzi Multigene Families
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João Luís Reis-Cunha, Anderson Coqueiro-dos-Santos, Samuel Alexandre Pimenta-Carvalho, Larissa Pinheiro Marques, Gabriela F. Rodrigues-Luiz, Rodrigo P. Baptista, Laila Viana de Almeida, Nathan Ravi Medeiros Honorato, Francisco Pereira Lobo, Vanessa Gomes Fraga, Lucia Maria da Cunha Galvão, Lilian Lacerda Bueno, Ricardo Toshio Fujiwara, Mariana Santos Cardoso, Gustavo Coutinho Cerqueira, and Daniella C. Bartholomeu
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Virology ,Microbiology - Abstract
Repetitive elements cause assembly fragmentation in complex eukaryotic genomes, limiting the study of their variability. The genome of Trypanosoma cruzi, the parasite that causes Chagas disease, has a high repetitive content, including multigene families. Although many T. cruzi multigene families encode surface proteins that play pivotal roles in host-parasite interactions, their variability is currently underestimated, as their high repetitive content results in collapsed gene variants. To estimate sequence variability and copy number variation of multigene families, we developed a read-based approach that is independent of gene-specific read mapping and
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- 2022
10. Serodiagnosis of leishmaniasis in asymptomatic and symptomatic dogs by use of the recombinant dynamin-1-like protein from Leishmania infantum: A preliminary study
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Williane Fernanda Siqueira, Mariana Santos Cardoso, Marianna de Carvalho Clímaco, Ana Luiza Teixeira Silva, Benjamin Heidt, Kasper Eersels, Bart van Grinsven, Daniella Castanheira Bartholomeu, Lilian Lacerda Bueno, Thomas Cleij, Ricardo Toshio Fujiwara, RS: FSE Sensor Engineering, Sensor Engineering, and Faculty of Science and Engineering
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LINKED-IMMUNOSORBENT-ASSAY ,Veterinary (miscellaneous) ,K-39 ,CHAGASI ,VISCERAL-LEISHMANIASIS ,KALA-AZAR ,DIAGNOSIS ,Recombinant antigens ,ANTIGENS ,CANINE LEISHMANIASIS ,Infectious Diseases ,Symptomatic dog ,DIRECT AGGLUTINATION-TEST ,Insect Science ,infantum ,INFECTION ,Parasitology ,Immunodiagnostic ,Asymptomatic dogs - Abstract
Visceral leishmaniasis (VL) is a fatal manifestation of an infection caused by intracellular protozoa of the Leishmania genus. In New World countries, VL is classified as a zoonotic disease with domestic dogs acting as its main reservoir. Asymptomatic dogs are as competent to transmit Leishmania to the vectors as symptomatic dogs, however current diagnostic tests are limited and present low sensitivity for this important group. The devel-opment of accurate tests is fundamental to the early diagnosis, treatment, and control of canine leishmaniasis. In this study, we investigated the use of a recombinant protein (dynamin-1-like protein, Dyn-1) from L. infantum, as a potential target antigen for leishmaniasis serodiagnosis in both symptomatic and asymptomatic dogs. The antigenic performance of the protein was evaluated by means of ELISA assays using sera from symptomatic (n = 25), asymptomatic (n = 34) and non-infected dogs (n = 36) using ELISA. In addition, sera from dogs experi-mentally infected with Trypanosoma cruzi (n = 49) and naturally infected with Babesia sp. (n = 8) were tested to evaluate possible cross-reactivity. A crude soluble antigen (CSA) of Leishmania was used as an antigen control and K39 and K26 were used as reference antigens because they are already widely used in commercial tests. rDyn-1-based assay showed the highest sensitivity (97%) compared to the antigens K39 (88%), K26 (86%) and crude extract (95%). The highest specificity among the tests was also obtained with the protein rDyn-1 (94%), compared with the other antigens K39 (81%), K26 (87%), and crude extract (77%). This study showed that the rDyn-1 ELISA assay was able to identify 100% of asymptomatic dogs, establishing its potential as a target for the diagnosis of canine leishmaniasis.
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- 2022
11. Nanoformulations with
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Jennifer, Ottino, Jaqueline Costa, Leite, Otoni Alves, Melo-Júnior, Marco Antonio Cabrera, González, Tatiane Furtado de, Carvalho, Giani Martins, Garcia, Maurício Azevedo, Batista, Patrícia, Silveira, Mariana Santos, Cardoso, Lilian Lacerda, Bueno, Ricardo Toshio, Fujiwara, Renato Lima, Santos, Paulo Ricardo de Oliveira, Paes, Denise, Silveira-Lemos, Olindo Assis, Martins-Filho, Alexsandro Sobreira, Galdino, Miguel Angel, Chávez-Fumagalli, Walderez Ornelas, Dutra, Vanessa Carla Furtado, Mosqueira, and Rodolfo Cordeiro, Giunchetti
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Leishmaniasis is a widespread vector-borne disease in Brazil, with
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- 2022
12. Impact of the COVID-19 pandemic on the actions of the Schistosomiasis Control Program in an endemic area in Northeastern Brazil
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Neiliane Medeiros Dantas, Lucas Almeida Andrade, Wandklebson Silva da Paz, Welde Natan Borges, Vanessa Gabriela Bernardino Barbosa, Diego Pereira Gonçalo da Hora, Carlos Eduardo da Silva, Rodrigo Feliciano do Carmo, Carlos Dornels Freire de Souza, Allan Dantas dos Santos, Flaviana Santos Wanderley, Deborah Aparecida Negrão-Corrêa, Ricardo Toshio Fujiwara, Márcio Bezerra-Santos, and Wagnner José Nascimento Porto
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Infectious Diseases ,Insect Science ,Veterinary (miscellaneous) ,Parasitology - Published
- 2023
13. Leishmanicidal Activity of the Volatile Oil of Piper macedoi
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Ricardo Toshio Fujiwara, Davyson de Lima Moreira, Gisele Lopes De Oliveira, Sebastião Rodrigo Ferreira, Rodrigo Guimaraes De Deus, Lúcia Pinheiro Santos Pimenta, Victor Neves Dos Santos, and Raquel Martins de Almeida
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Dillapiole ,Piper ,Chromatography ,biology ,Apiole ,Leishmaniasis ,Piperaceae ,biology.organism_classification ,medicine.disease ,chemistry.chemical_compound ,Camphor ,chemistry ,medicine ,Gas chromatography ,General Pharmacology, Toxicology and Pharmaceutics ,Leishmania infantum - Abstract
Leishmaniasis are zoonosis caused by more than 20 species of protozoa. The treatment is carried out with pentavalent antimonials (Sb5+) and amphotericin B, neither of which is fully effective. In the present study, the leishmanicidal activity and the cytotoxic potential of the volatile oil of Piper macedoi Yunck., Piperaceae, were evaluated. Extraction was achieved by hydrodistillation in a Clevenger-type apparatus, followed by chemical analysis using gas chromatography coupled to a flame ionization detector and gas chromatography coupled to mass spectrometry. The in vitro test was performed against amastigotes of Leishmania infantum (BH46) and cytotoxic activity was evaluated against canine macrophages (DH82), human hepatoma cells (HepG2), and monkey renal cells (BGM). P. macedoi proved to be effective with leishmanicidal activity against amastigotes of L. infantum with an IC50 of 222.4 μg/ml. The cytotoxic concentrations (CC50) were 316.8, 194.8, and 300.6 μg/ml to DH82, HepG2, and BGM, respectively. The four major substances found in the volatile oil were the arylpropanoids apiole (14.89%) and dillapiole (11.54%), and the monoterpenes 1,8-cineole (14.08%) and camphor (10.19%).
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- 2021
14. Immunological underpinnings of Ascaris infection, reinfection and co-infection and their associated co-morbidities
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Ana Clara Gazzinelli-Guimarães, Lilian Lacerda Bueno, Luisa Md Magalhães, Ricardo Toshio Fujiwara, Flaviane Vieira-Santos, Denise Silva Nogueira, Pedro Henrique Gazzinelli-Guimarães, Fabrício Marcus Silva Oliveira, and Lucas Kraemer
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0301 basic medicine ,biology ,Ascaris ,030231 tropical medicine ,Tropical disease ,Context (language use) ,biology.organism_classification ,medicine.disease ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Infectious Diseases ,Immune system ,Ascariasis ,Immunology ,Coinfection ,medicine ,Animal Science and Zoology ,Parasitology ,Co morbidity ,Co infection - Abstract
Human ascariasis is the most common and prevalent neglected tropical disease and is estimated that ~819 million people are infected around the globe, accounting for 0.861 million years of disability-adjusted life years in 2017. Even with the existence of highly effective drugs, the constant presence of infective parasite eggs in the environment contribute to a high reinfection rate after treatment. Due to its high prevalence and broad geographic distribution Ascaris infection is associated with a variety of co-morbidities and co-infections. Here, we provide data from both experimental models and humans studies that illustrate how complex is the interaction of Ascaris with the host immune system, especially, in the context of reinfections, co-infections and associated co-morbidities.
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- 2021
15. IL-17RA receptor signaling contributes to lung inflammation and parasite burden during Toxocara canis infection in mice
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Thaís Leal-Silva, Camila de Almeida Lopes, Flaviane Vieira-Santos, Fabrício Marcus Silva Oliveira, Lucas Kraemer, Luiza de Lima Silva Padrão, Chiara Cássia Oliveira Amorim, Jorge Lucas Nascimento Souza, Remo Castro Russo, Ricardo Toshio Fujiwara, Luisa Mourão Dias Magalhães, and Lilian Lacerda Bueno
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Immunology ,Immunology and Allergy - Abstract
IL-17 is a cytokine produced by innate and acquired immunity cells that have an action against fungi and bacteria. However, its action in helminth infections is unclear, including in Toxocara canis infection. Toxocariasis is a neglected zoonosis representing a significant public health problem with an estimated seroprevalence of 19% worldwide. In the present study, we describe the immunopathological action of IL-17RA in acute T. canis infection. C57BL/6j (WT) and IL-17RA receptor knockout (IL-17RA-/-) mice were infected with 1000 T. canis eggs. Mice were evaluated 3 days post-infection for parasite load and white blood cell count. Lung tissue was harvested for histopathology and cytokine expression. In addition, we performed multiparametric flow cytometry in the BAL and peripheral blood, evaluating phenotypic and functional changes in myeloid and lymphoid populations. We showed that IL-17RA is essential to control larvae load in the lung; however, IL-17RA contributed to pulmonary inflammation, inducing inflammatory nodular aggregates formation and presented higher pulmonary IL-6 levels. The absence of IL-17RA was associated with a higher frequency of neutrophils as a source of IL-4 in BAL, while in the presence of IL-17RA, mice display a higher frequency of alveolar macrophages expressing the same cytokine. Taken together, this study indicates that neutrophils may be an important source of IL-4 in the lungs during T. canis infection. Furthermore, IL-17/IL-17RA axis is important to control parasite load, however, its presence triggers lung inflammation that can lead to tissue damage.
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- 2022
16. Organosulfur Compounds: Leishmanicidal and Nematicidal Activities of Mansoa alliacea Leaf Essential Oil on Leishmania amazonensis and Caenorhabditis elegans
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Francisco das Chagas do Nascimento, Jonkácio Almeida de Melo, Luciana Poty Manso dos Santos, Ricardo Toshio Fujiwara, Raquel Martins de Almeida, Sebastião Rodrigo Ferreira, and Leandro da Silva Nascimento
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General Chemistry - Abstract
Doencas parasitarias representam um serio problema global e afetam diretamente a saude e a qualidade de vida da populacao. Mansoa alliacea e uma bignoniaceae encontrada principalmente nas matas brasileiras, conhecida como “cipo-de-alho” devido ao forte odor de alho. E uma planta utilizada na medicina popular, com relatos de diversas atividades biologicas (anti-inflamatoria, antifungica, antimicrobiana e antioxidante). O objetivo deste trabalho e caracterizar o oleo essencial de M. alliacea e sua avaliacao inedita das atividades nematicida e leishmanicida contra Leishmania amazonensis e Caenorhabditis elegans. O oleo essencial foi obtido por hidrodestilacao, sua composicao quimica foi analisada por GC-MS e quantificada por GC-FID. A atividade leishmanicida foi realizada em amastigotas intracelulares de L. amazonensis usando macrofagos caninos DH82, a atividade foi medida usando Enzyme Linked Immuno Sorbent Assay (ELISA). A atividade nematicida foi avaliada em larvas do modelo nematoide C. elegans por meio da avaliacao da motilidade. A citotoxicidade do oleo foi avaliada por ensaio colorimetrico de sal de tetrazolio (MTT) em diferentes concentracoes. Os farmacos utilizados como controle positivo foram: antimonio tetravalente na atividade leishmanicida e ivermectina na atividade nematicida. A analise de GC-MS revelou o perfil organossulfurado do oleo, tendo como majoritarios: metil alil dissulfeto (2,4%), dialil tetrassulfeto (3,4%) dialil dissulfeto (36,6%), dialil trissulfeto (53,3%). O oleo apresentou excelente atividade leishmanicida, IC50 de 9,4 μg/mL, valor 5 vezes menor que o antimonio (III) (IC50 53 μg/mL). A dose citotoxica (CC50) do oleo foi 90,7 μg/mL, enquanto o antimonio foi CC50=1,7 μg/mL. O indice de seletividade (razao entre CC50/IC50) do oleo foi 10 e o antimonio (III) foi 0,03. O oleo tambem foi eficiente em paralisar as larvas de C. elegans (IC50=113 µg/mL), na ordem de 94,65% das larvas, superior ao farmaco de referencia, ivermectina (92,2 %), na mesma concentracao. Os resultados ineditos, aqui apresentados, demonstram o potencial de M. alliacea no desenvolvimento de novos farmacos para o tratamento de leishmanioses e nematoides.
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- 2021
17. Vaccination with chimeric protein induces protection in murine model against ascariasis
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Maria Elena Bottazzi, Joseane Camilla de Castro, Bin Zhan, Denise Silva Nogueira, Fabrício Marcus Silva Oliveira, Daniella Castanheira Bartholomeu, Laila Viana de Almeida, Lilian Lacerda Bueno, João Luís Reis-Cunha, Peter J. Hotez, Ricardo Toshio Fujiwara, Luísa Mourão Dias Magalhães, and Mariana Santos Cardoso
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Recombinant Fusion Proteins ,030231 tropical medicine ,Monophosphoryl Lipid A ,Epitope ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Antigen ,medicine ,Animals ,030212 general & internal medicine ,Ascaris suum ,B cell ,Ascariasis ,Mice, Inbred BALB C ,General Veterinary ,General Immunology and Microbiology ,biology ,Ascaris ,Vaccination ,Public Health, Environmental and Occupational Health ,biology.organism_classification ,Fusion protein ,Virology ,Disease Models, Animal ,Infectious Diseases ,medicine.anatomical_structure ,Antigens, Helminth ,Molecular Medicine ,Female - Abstract
An estimated 400 million people are infected by parasites of the genus Ascaris and the existing control measures are inefficient. Vaccine development using B cell antigens is a promising strategy for increased protection against this parasite. The present study aimed at developing a chimeric protein capable of conferring protection against infection by Ascaris sp. For this purpose, we performed B-cell epitope predictions on previously described vaccine candidate proteins from Ascaris suum and the corresponding peptides were used to construct a chimeric protein. Female BALB / c mice were immunized subcutaneously in three doses at 10 day intervals with a vaccine formulation comprised of the chimeric protein together with monophosphoryl lipid A (MPLA). Control groups included protein alone, MPLA, or PBS. After challenge infection, animals vaccinated with chimeric protein plus MPLA showed a reduction of 73.54% of larval load in the lung compared to control group animals. Animals immunized with chimeric protein plus MPLA also display higher IgG response and a reduction in lung inflammation. Our study highlights how chimeric proteins containing more than one B cell epitope can enhance immune protection against helminthic infection and offer new approaches to the development of Ascaris vaccines.
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- 2021
18. Vaccination with Formulation of Nanoparticles Loaded with Leishmania amazonensis Antigens Confers Protection against Experimental Visceral Leishmaniasis in Hamster
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Marco Antonio Cabrera González, Ana Alice Maia Gonçalves, Jennifer Ottino, Jaqueline Costa Leite, Lucilene Aparecida Resende, Otoni Alves Melo-Júnior, Patrícia Silveira, Mariana Santos Cardoso, Ricardo Toshio Fujiwara, Lilian Lacerda Bueno, Renato Lima Santos, Tatiane Furtado de Carvalho, Giani Martins Garcia, Paulo Ricardo de Oliveira Paes, Alexsandro Sobreira Galdino, Miguel Angel Chávez-Fumagalli, Marília Martins Melo, Denise Silveira-Lemos, Olindo Assis Martins-Filho, Walderez Ornelas Dutra, Vanessa Carla Furtado Mosqueira, and Rodolfo Cordeiro Giunchetti
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Pharmacology ,Infectious Diseases ,Drug Discovery ,Immunology ,visceral leishmaniasis ,nanotechnology ,vaccine ,Pharmacology (medical) - Abstract
Visceral leishmaniasis (VL) is a fatal disease caused by the protozoa Leishmania infantum for which dogs are the main reservoirs. A vaccine against canine visceral leishmaniasis (CVL) could be an important tool in the control of human and CVL by reducing the infection pressure of L. infantum. Despite the CVL vaccine available on the market, the Brazilian Ministry of Health did not implement the use of it in their control programs. In this sense, there is an urgent need to develop more efficient vaccines. In this study, the association between two polymeric nanoformulations, (poly (D, L-lactic) acid (PLA) polymer) loading Leishmania amazonensis antigens, was evaluated as a potential immunobiological agent against VL using golden hamsters as an experimental model. The results indicated that no significant adverse reactions were observed in animals vaccinated with LAPSmP. LAPSmP presented similar levels of total anti-Leishmania IgG as compared to LAPSmG. The LAPSmP and LAPSmG groups showed an intense reduction in liver and spleen parasitic load by qPCR. The LAPSmP and LAPSmG vaccines showed exceptional results, indicating that they may be promising candidates as a VL vaccine.
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- 2023
19. Visceral leishmaniasis in a recent transmission region: 27.4% infectivity rate among seronegative dogs
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Carolina Novato Gondim, Sidney de Almeida Ferreira, Beatriz Ketelin Sousa Vasconcelos, Flademir Wouters, Ricardo Toshio Fujiwara, Joseane Camilla de Castro, and Joziana Muniz de Paiva Barçante
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Infectious Diseases ,Animal Science and Zoology ,Parasitology - Abstract
American visceral leishmaniasis (VL) is a parasitic disease whose main domestic reservoir in the urban environment is dog and is considered one of the most important zoonoses in the context of public health. Serological tests are typically used for the diagnostic screening of the disease. This study aimed to analyse the performance of different methodologies used in the diagnosis of VL in dogs sampled from a recent transmission area. The sample consisted of 52 dogs separated into groups based on the absence and presence of clinical signs of VL. The following serological techniques were carried out: the DPP® rapid test (RT), the ALERE® RT and an RT and immunoenzymatic assay with a recently developed protein (rKDDR-plus). In addition, molecular techniques were carried out with conjunctival swabs, and bone marrow aspirate samples and parasitological samples were obtained directly from bone marrow aspirates. It was concluded that 27.4% of seronegative dogs were infected, but the serological tests, used as screening tests, showed unsatisfactory sensitivity results (average: 51.2%) for dogs without clinical signs. It was suggested that polymerase chain reaction with conjunctival swabbing be used as a screening test for dogs without clinical signs, as this is a non-invasive collection technique with high-sensitivity values.
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- 2022
20. Genetic Background Affects the Mucosal Secretory IgA Levels, Parasite Burden, Lung Inflammation, and Mouse Susceptibility toAscaris suumInfection
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Ana Maria Caetano Faria, Soraya Gaze, Chiara Cássia Oliveira Amorim, Lucas Kraemer, Matheus Silvério Mattos, Natália Gabriela Silva Pinheiro, Ana Clara Gazzinelli-Guimarães, Ricardo Toshio Fujiwara, Nathalia Maria Resende, Lilian Lacerda Bueno, Luciana Maria Oliveira, Remo Castro Russo, Fernando Sérgio Barbosa, Denise Silva Nogueira, and Ricardo M. Geraldi
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Lung ,biology ,Immunology ,Tropical disease ,Inflammation ,biology.organism_classification ,medicine.disease ,Microbiology ,Infectious Diseases ,medicine.anatomical_structure ,Immune system ,Intestinal mucosa ,Ascariasis ,medicine ,Helminths ,Parasitology ,medicine.symptom ,Ascaris suum - Abstract
Ascariasis is a neglected tropical disease, widespread in the world and causing important socioeconomic impacts. The presence of various stages of worm development in the pulmonary and intestinal mucosa induces a humoral and cellular immune response. However, although there is much evidence of the protective role of mucosal immunity against various pathogens, including helminthes, there is still a gap in the knowledge about the immune response and the mechanisms of action that are involved in protection against diseases, especially in the initial phase of ascariasis. Then, the aim of this study was to evaluate the kinetic aspects of the immune parasitological parameters in intestinal and pulmonary mucosa in male mice with early ascariasis. Therefore, two mice strains showed a different susceptibility to ascariasis (BALB/c and C57BL6/j), when experimentally infected with 2,500 infective eggs of Ascaris suum from time-point 0 and the immune parasitological parameters evaluated each two days after infection, during the period of 12 days. The results were suggestive of a synergetic action of intestinal and pulmonary SIgA contributing for the protection against early ascariasis by reducing the amount of migrating larval as well as the influx of leukocytes in the lung and the consequent impair of the pulmonary capacity.
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- 2022
21. Tissue eosinophilia correlates with mice susceptibility, granuloma formation, and damage during
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Thaís, Leal-Silva, Camila de Almeida, Lopes, Flaviane, Vieira-Santos, Fabrício Marcus Silva, Oliveira, Lucas, Kraemer, Luiza de Lima Silva, Padrão, Chiara Cássia Oliveira, Amorim, Jorge Lucas Nascimento, Souza, Fernando Sérgio, Barbosa, Milene Alvarenga, Rachid, Remo Castro, Russo, Ricardo Toshio, Fujiwara, and Lilian Lacerda, Bueno
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- 2022
22. IL-17RA receptor signaling contributes to lung inflammation and parasite burden during
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Thaís, Leal-Silva, Camila de Almeida, Lopes, Flaviane, Vieira-Santos, Fabrício Marcus Silva, Oliveira, Lucas, Kraemer, Luiza de Lima Silva, Padrão, Chiara Cássia Oliveira, Amorim, Jorge Lucas Nascimento, Souza, Remo Castro, Russo, Ricardo Toshio, Fujiwara, Luisa Mourão Dias, Magalhães, and Lilian Lacerda, Bueno
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Mice, Inbred C57BL ,Mice ,Receptors, Interleukin-17 ,Toxocariasis ,Interleukin-17 ,Animals ,Cytokines ,Toxocara canis ,Interleukin-4 ,Pneumonia - Abstract
IL-17 is a cytokine produced by innate and acquired immunity cells that have an action against fungi and bacteria. However, its action in helminth infections is unclear, including in
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- 2022
23. Diagnostic application of sensitive and specific phage-exposed epitopes for visceral leishmaniasis and human immunodeficiency virus coinfection
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Ricardo Toshio Fujiwara, Vívian T. Martins, Amanda S. Machado, Grasiele S.V. Tavares, Isabela A. P. Gonçalves, Vanessa Gomes Fraga, Fernanda F. Ramos, Daniela P. Lage, Thiago A.R. Reis, Fernanda Ludolf, João A. Oliveira-da-Silva, Nathalia Sernizon Guimarães, Thaís T.O. Santos, Camila S. Freitas, Ana C. S. Dias, Raquel S. Bandeira, Lilian Lacerda Bueno, Ana Thereza Chaves, Eduardo A.F. Coelho, Patrícia Terra Alves, Luiz Ricardo Goulart, Gláucia Fernandes Cota, Unaí Tupinambás, Manoel Otávio da Costa Rocha, and Miguel A. Chávez-Fumagalli
- Subjects
Coinfection ,Human immunodeficiency virus (HIV) ,HIV ,HIV Infections ,Biology ,medicine.disease ,medicine.disease_cause ,Virology ,Epitope ,Epitopes ,Infectious Diseases ,Visceral leishmaniasis ,medicine ,Humans ,Leishmaniasis, Visceral ,Animal Science and Zoology ,Parasitology ,Bacteriophages - Abstract
The diagnosis of visceral leishmaniasis (VL) has improved with the search of novel antigens; however, their performance is limited when samples from VL/human immunodeficiency virus (HIV)-coinfected patients are tested. In this context, studies conducted to identify more suitable antigens to detect both VL and VL/HIC coinfection cases should be performed. In the current study, phage display was performed using serum samples from healthy subjects and VL, HIV-infected and VL/HIV-coinfected patients; aiming to identify novel phage-exposed epitopes to be evaluated with this diagnostic purpose. Nine non-repetitive and valid sequences were identified, synthetized and tested as peptides in enzyme-linked immunosorbent assay experiments. Results showed that three (Pep2, Pep3 and Pep4) peptides showed excellent performance to diagnose VL and VL/HIV coinfection, with 100% sensitivity and specificity values. The other peptides showed sensitivity varying from 50.9 to 80.0%, as well as specificity ranging from 60.0 to 95.6%. Pep2, Pep3 and Pep4 also showed a potential prognostic effect, since specific serological reactivity was significantly decreased after patient treatment. Bioinformatics assays indicated that Leishmania trypanothione reductase protein was predicted to contain these three conformational epitopes. In conclusion, data suggest that Pep2, Pep3 and Pep4 could be tested for the diagnosis of VL and VL/HIV coinfection.
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- 2022
24. Effect on cellular recruitment and the innate immune response by combining saponin, monophosphoryl lipid-A and Incomplete Freund’s Adjuvant with Leishmania (Viannia) braziliensis antigens for a vaccine formulation
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Paula Melo de Abreu Vieira, Maria Norma Melo, Rodolfo Cordeiro Giunchetti, Alexandre Barbosa Reis, Cláudia Martins Carneiro, Fernando Augusto Siqueira Mathias, Ricardo Toshio Fujiwara, Andréa Teixeira-Carvalho, Nádia das Dores Moreira, Juliana Vitoriano-Souza, Rodrigo Dian de Oliveira Aguiar-Soares, Bruno Mendes Roatt, and Rory Cristiane Fortes de Brito
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Male ,medicine.medical_treatment ,Freund's Adjuvant ,030231 tropical medicine ,Antibodies, Protozoan ,Monophosphoryl Lipid A ,Antigens, Protozoan ,chemical and pharmacologic phenomena ,Biology ,Leishmania braziliensis ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Adjuvants, Immunologic ,Antigen ,medicine ,Animals ,030212 general & internal medicine ,IL-2 receptor ,Leishmaniasis Vaccines ,Innate immune system ,General Veterinary ,General Immunology and Microbiology ,Immunogenicity ,Public Health, Environmental and Occupational Health ,Saponins ,Acquired immune system ,Lipids ,Immunity, Innate ,Lipid A ,Infectious Diseases ,Antibody Formation ,Immunology ,Leishmaniasis, Visceral ,Molecular Medicine ,Adjuvant - Abstract
The poor immunogenicity displayed by some antigens has encouraged the development of strategies to improve the immune response and safety of vaccine candidates, resulting in an intense search for substances that potentiate vaccine response. Adjuvants have these properties helping vaccine candidates to induce a strong, durable, and fast immune response. In this study, we evaluated the specific immune response of adjuvants alone, Saponin (SAP), Incomplete Freund’s Adjuvant (IFA) and Monophosphoryl lipid-A SE (MPL-SE®) and in combination with total antigen of L. braziliensis (LB): LBSAP, LBIFA and LBMPL. The specific immune response induced by these compositions demonstrated that they were powerfully immunogenic, increasing cellular infiltration in the skin. Draining lymph nodes cultures showed that LBIFA and LBMPL have higher ability to increase the capacity of APCs to present antigens, with increased frequency of CD11c+CD86+ cells. SAP, MPL, LBSAP, LBIFA and LBMPL could activate lymphocytes increasing expression of CD69 and CD25. LBSAP group was an excellent inducer of pro-inflammatory cytokines at 24 h. At 48 h, higher cytokines production was observed in IFA, LBIFA, MPL and LBMPL groups. Our data demonstrate that LBSAP and LBMPL are potential formulations to be tested in other experimental models. Also, the data obtained could expand the knowledge about immune response after sensitization and also contribute to the development of safe, immunogenic and effective vaccines.
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- 2019
25. ASCVac-1, a Multi-Peptide Chimeric Vaccine, Protects Mice Against Ascaris suum Infection
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Ana Clara Gazzinelli-Guimarães, Denise Silva Nogueira, Chiara Cássia Oliveira Amorim, Fabrício Marcus Silva Oliveira, Anderson Coqueiro-Dos-Santos, Samuel Alexandre Pimenta Carvalho, Lucas Kraemer, Fernando Sérgio Barbosa, Vanessa Gomes Fraga, Flaviane Vieira Santos, Joseane Camilla de Castro, Remo Castro Russo, Milena Apetito Akamatsu, Paulo Lee Ho, Maria Elena Bottazzi, Peter J. Hotez, Bin Zhan, Daniella Castanheira Bartholomeu, Lilian Lacerda Bueno, and Ricardo Toshio Fujiwara
- Subjects
Protozoan Vaccines ,Immunology ,Neglected Diseases ,Vaccine Efficacy ,RC581-607 ,epitopes ,chimera ,ascariasis ,Mice ,Th2 Cells ,Antigens, Helminth ,vaccine ,Vaccines, Subunit ,Animals ,Humans ,Immunology and Allergy ,Female ,vaccinology ,Immunologic diseases. Allergy ,Lung ,Ascaris suum ,Original Research - Abstract
Control of human ascariasis, the most prevalent neglected tropical disease globally affecting 450 million people, mostly relies on mass drug administration of anthelmintics. However, chemotherapy alone is not efficient due to the high re-infection rate for people who live in the endemic area. The development of a vaccine that reduces the intensity of infection and maintains lower morbidity should be the primary target for infection control. Previously, our group demonstrated that immunization with crude Ascaris antigens in mice induced an IgG-mediated protective response with significant worm reduction. Here, we aimed to develop a multipeptide chimera vaccine based on conserved B-cell epitopes predicted from 17 common helminth proteomes using a bioinformatics algorithm. More than 480 B-cell epitopes were identified that are conserved in all 17 helminths. The Ascaris-specific epitopes were selected based on their reactivity to the pooled sera of mice immunized with Ascaris crude antigens or infected three times with A. suum infective eggs. The top 35 peptides with the strongest reactivity to Ascaris immune serum were selected to construct a chimeric antigen connected in sequence based on conformation. This chimera, called ASCVac-1, was produced as a soluble recombinant protein in an Escherichia coli expression system and, formulated with MPLA, was used to immunize mice. Mice immunized with ASCVac-1/MPLA showed around 50% reduced larvae production in the lungs after being challenged with A. suum infective eggs, along with significantly reduced inflammation and lung tissue/function damage. The reduced parasite count and pathology in infected lungs were associated with strong Th2 immune responses characterized by the high titers of antigen-specific IgG and its subclasses (IgG1, IgG2a, and IgG3) in the sera and significantly increased IL-4, IL-5, IL-13 levels in lung tissues. The reduced IL-33 titers and stimulated eosinophils were also observed in lung tissues and may also contribute to the ASCVac-1-induced protection. Taken together, the preclinical trial with ASCVac-1 chimera in a mouse model demonstrated its significant vaccine efficacy associated with strong IgG-based Th2 responses, without IgE induction, thus reducing the risk of an allergic response. All results suggest that the multiepitope-based ASCVac-1 chimera is a promising vaccine candidate against Ascaris sp. infections.
- Published
- 2021
26. Genetic Background Affects the Mucosal Secretory IgA Levels, Parasite Burden, Lung Inflammation, and Mouse Susceptibility to
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Luciana Maria, Oliveira, Denise Silva, Nogueira, Ricardo Marcelo, Geraldi, Fernando Sérgio, Barbosa, Chiara Cássia Oliveira, Amorim, Ana Clara, Gazzinelli-Guimarães, Nathália Maria, Resende, Natália, Pinheiro-Rosa, Lucas Rocha, Kraemer, Matheus Silvério, Mattos, Lilian Lacerda, Bueno, Ana Maria Caetano, Faria, Remo Castro, Russo, Soraya, Gaze, and Ricardo Toshio, Fujiwara
- Subjects
Male ,Mice, Inbred C57BL ,Swine Diseases ,Ascariasis ,Mice ,Host Response and Inflammation ,Swine ,Immunoglobulin A, Secretory ,Animals ,Parasites ,Pneumonia ,Genetic Background ,Ascaris suum - Abstract
Ascariasis is a neglected tropical disease that is widespread in the world and has important socioeconomic impacts. The presence of various stages of worm development in the pulmonary and intestinal mucosae induces a humoral and cellular immune response. However, although there is much evidence of the protective role of mucosal immunity against various pathogens, including helminths, there is still a gap in the knowledge about the immune response and the mechanisms of action that are involved in protection against diseases, especially in the initial phase of ascariasis. Thus, the aim of this study was to evaluate the kinetic aspects of the immune parasitological parameters in intestinal and pulmonary mucosae in male mice with early ascariasis. Therefore, two mouse strains that showed different susceptibilities to ascariasis (BALB/c and C57BL/6J) when experimentally infected with 2,500 infective eggs of Ascaris suum from time point 0 were examined: the immune parasitological parameters were evaluated each 2 days after infection over a period of 12 days. The results were suggestive of a synergetic action of intestinal and pulmonary secretory IgA (S-IgA) contributing to protection against early ascariasis by reducing the amount of migrating larvae as well as the influx of leukocytes in the lung and the consequent impairment of pulmonary capacity.
- Published
- 2021
27. Intestinal polyparasitism and levels of mucosal anthelmintic SIgA in children from endemic areas in Northeastern Brazil
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Silvio Santana Dolabella, Ricardo Toshio Fujiwara, Yrna Lorena Matos de Oliveira, Vitor L Sá, Ricardo M. Geraldi, Lilian Lacerda Bueno, Yvanna L.D.C. Oliveira, Carina S. Pinheiro, Anne Caroline Santos Ramos, Luciana Maria Oliveira, and Gabriela F Andrade
- Subjects
Saliva ,Immunology ,Population ,Helminthiasis ,Asymptomatic ,Feces ,Soil ,fluids and secretions ,Immune system ,parasitic diseases ,medicine ,Prevalence ,Helminths ,Animals ,Humans ,Anthelmintic ,Intestinal Diseases, Parasitic ,education ,Child ,Anthelmintics ,education.field_of_study ,biology ,biology.organism_classification ,Mucosal immunology ,Immunoglobulin A, Secretory ,Protozoa ,Parasitology ,medicine.symptom ,Brazil ,medicine.drug - Abstract
BACKGROUND Interactions between parasites during coinfections are often complex and can impact immunization and treatment programs, as well as disease outcomes and morbidity. However, little is known about these interactions and the mechanisms involved. METHODS In this study, a coproparasitological survey was carried out in school-age children living in endemic areas of parasitic infection in the state of Sergipe, in Northeastern Brazil. Anti-helminth-specific and total secretory immunoglobulin-A (SIgA) levels were measured in stool and saliva samples and were compared in children presenting monoparasitism, polyparasitism (helminths and/or intestinal protozoa), and no infections. RESULTS The survey showed that protozoa were more prevalent than helminths, and that there was a high frequency of polyparasitism in the studied population, mainly from combinations of protozoan species. Although less frequent, combinations between species of protozoa and helminths were also observed. The levels of salivary SIgA in these co-infected individuals were lower than the average observed in infections with helminths alone. CONCLUSIONS Although the children participating in this survey were asymptomatic, and it was, therefore, not possible to evaluate the impact of salivary SIgA reduction on the diseases, the study highlights the need for further investigations of coinfections by intestinal parasites and the effects on immune response induced by the interactions between different parasites.
- Published
- 2021
28. Reduced vitamin D receptor (VDR) and cathelicidin antimicrobial peptide (CAMP) gene expression contribute to the maintenance of inflammatory immune response in leprosy patients
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Ana Laura Grossi de Oliveira, Ana Thereza Chaves, Mariana Santos Cardoso, Guilherme Rafael Gomide Pinheiro, Douglas Eulálio Antunes, Maria Aparecida de Faria Grossi, Sandra Lyon, Lilian Lacerda Bueno, Manoel Otávio da Costa Rocha, Cristiane Alves da Silva Menezes, and Ricardo Toshio Fujiwara
- Subjects
Immunology ,Interleukin-17 ,Immunity ,Gene Expression ,Leprostatic Agents ,Microbiology ,Mycobacterium leprae ,Infectious Diseases ,Cathelicidins ,Leprosy ,Cytokines ,Humans ,Interleukin-2 ,Receptors, Calcitriol ,Drug Therapy, Combination ,Vitamin D ,Antimicrobial Peptides ,Antimicrobial Cationic Peptides ,Transcription Factors - Abstract
Leprosy is an infectious disease influenced by genetic, immunological, and environmental factors. Reduced gene expressions may be associated with the immunological response pattern and leprosy susceptibility. We investigated the direct and indirect effects of Vitamin D Receptor (VDR) and Cathelicidin Antimicrobial Peptide (CAMP) gene expressions on the serum levels of vitamin D, Cathelicidin, and cytokines in newly-diagnosed leprosy patients and post-six-months of multidrug therapy (MDT). Thirty-four leprosy patients were assessed, paucibacillary (PB; n = 14) and multibacillary (MB; n = 20) cases, untreated or having received six months of MDT, 18 healthy controls, and 25 household contacts. VDR and CAMP gene expression levels were strongly correlated to some important cytokines in both, untreated leprosy patients (PB, r = 0.9319; MB, r = 0.9569) and patients who had undergone MDT (PB, r = 0.9667; MB, r = 0.9569). We observed that both gene expressions directly influenced IL-2, IFN-γ, and IL-17F serum levels in leprosy patients compared to the household contacts and healthy individuals. VDR and CAMP gene expressions induced a persistent inflammatory response in PB and MB leprosy patients, even after six months of MDT, to fight the Mycobacterium leprae infection. Due to the persistent inflammatory profile, multidrug therapy is suggested to be maintained for more than six months, especially for MB patients. Vitamin D supplementation is recommended from the onset as a transcription factor to improve VDR and CAMP gene expression in leprosy patients.
- Published
- 2021
29. Nanoformulations with Leishmania braziliensis Antigens Triggered Controlled Parasite Burden in Vaccinated Golden Hamster (Mesocricetus auratus) against Visceral Leishmaniasis
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Jennifer Ottino, Jaqueline Costa Leite, Otoni Alves Melo-Júnior, Marco Antonio Cabrera González, Tatiane Furtado de Carvalho, Giani Martins Garcia, Maurício Azevedo Batista, Patrícia Silveira, Mariana Santos Cardoso, Lilian Lacerda Bueno, Ricardo Toshio Fujiwara, Renato Lima Santos, Paulo Ricardo de Oliveira Paes, Denise Silveira-Lemos, Olindo Assis Martins-Filho, Alexsandro Sobreira Galdino, Miguel Angel Chávez-Fumagalli, Walderez Ornelas Dutra, Vanessa Carla Furtado Mosqueira, and Rodolfo Cordeiro Giunchetti
- Subjects
Pharmacology ,Infectious Diseases ,visceral leishmaniasis ,polymeric nanoparticle ,vaccine ,hamster ,pre-clinical trial ,Drug Discovery ,Immunology ,Pharmacology (medical) - Abstract
Leishmaniasis is a widespread vector-borne disease in Brazil, with Leishmania (Leishmania) infantum as the primary etiological agent of visceral leishmaniasis (VL). Dogs are considered the main reservoir of this parasite, whose treatment in Brazil is restricted to the use of veterinary medicines, which do not promote a parasitological cure. Therefore, efficient vaccine development is the best approach to Canine Visceral Leishmaniasis (CVL) control. With this in mind, this study used hamsters (Mesocricetus auratus) as an experimental model in an anti-Leishmania preclinical vaccine trial to evaluate the safety, antigenicity, humoral response, and effects on tissue parasite load. Two novel formulations of nanoparticles made from poly(D, L-lactic) acid (PLA) polymer loading Leishmania braziliensis crude antigen (LB) exhibiting two different particle sizes were utilized: LBPSmG (570 nm) and LBPSmP (388 nm). The results showed that the nanoparticles were safe and harmless to hamsters and were antigenic with the induction in LBSap, LBPSmG, and LBPSmG groups of total anti-Leishmania IgG antibodies 30 days after challenge, which persists 200 days in LBSap and LBPSmP. At the same time, a less pronounced hepatosplenomegaly in LBSap, LBPSmG, and LBPSmP was found when compared to control groups, as well as a less pronounced inflammatory infiltrate and granuloma formation in the spleen. Furthermore, significant reductions of 84%, 81%, and 90% were observed in spleen parasite burden accessed by qPCR in the LBSap, LBPSmG, and LBPSmP groups, respectively. In this way, LBSap, LBPSmG, and LBPSmP formulations showed better results in vaccinated and L. infantum-challenged animals in further reducing parasitic load in the spleen and attenuating lesions in liver and splenic tissues. This results in safe, harmless nanoformulation vaccines with significant immunogenic and infection control potential. In addition, animals vaccinated with LBPSmP had an overall reduction in parasite burden in the spleen, indicating that a smaller nanoparticle could be more efficient in targeting antigen-presenting cells.
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- 2022
30. Inhalation of dimethyl fumarate-encapsulated solid lipid nanoparticles attenuate clinical signs of experimental autoimmune encephalomyelitis and pulmonary inflammatory dysfunction in mice
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Bárbara Fernandes Pinto, Lorena Natasha Brito Ribeiro, Gisela Bevilacqua Rolfsen Ferreira da Silva, Camila Simões Freitas, Lucas Kraemer, Fabrício Marcus Silva Oliveira, Marianna Carvalho Clímaco, Flávio Afonso Gonçalves Mourão, Gabryella Soares Pinheiro dos Santos, Samantha Ribeiro Béla, Isabella Luísa da Silva Gurgel, Fábio de Lima Leite, Anselmo Gomes de Oliveira, Maura Regina Silva da Páscoa Vilela, Onésia Cristina Oliveira-Lima, Frederico Marianetti Soriani, Ricardo Toshio Fujiwara, Alexander Birbrair, Remo Castro Russo, Juliana Carvalho-Tavares, Universidade Federal de Minas Gerais (UFMG), Universidade Federal de São Carlos (UFSCar), Universidade Estadual Paulista (UNESP), and Federal University of Goias (UFG)
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Mice, Inbred C57BL ,Disease Models, Animal ,Encephalomyelitis, Autoimmune, Experimental ,Multiple Sclerosis ,Dimethyl Fumarate ,Administration, Inhalation ,Liposomes ,Animals ,Nanoparticles ,Female ,General Medicine ,Pneumonia ,Immunosuppressive Agents - Abstract
Made available in DSpace on 2022-04-29T08:38:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-01-01 Rationale: The FDA-approved Dimethyl Fumarate (DMF) as an oral drug for Multiple Sclerosis (MS) treatment based on its immunomodulatory activities. However, it also caused severe adverse effects mainly related to the gastrointestinal system. Objective: Investigated the potential effects of solid lipid nanoparticles (SLNs) containing DMF, administered by inhalation on the clinical signs, central nervous system (CNS) inflammatory response, and lung function changes in mice with experimental autoimmune encephalomyelitis (EAE). Materials and methods: EAE was induced using MOG35-55 peptide in female C57BL/6J mice and the mice were treated via inhalation with DMF-encapsulated SLN (CTRL/SLN/DMF and EAE/SLN/DMF), empty SLN (CTRL/SLN and EAE/SLN), or saline solution (CTRL/saline and EAE/saline), every 72 h during 21 days. Results: After 21 days post-induction, EAE mice treated with DMF-loaded SLN, when compared with EAE/saline and EAE/SLN, showed decreased clinical score and weight loss, reduction in brain and spinal cord injury and inflammation, also related to the increased influx of Foxp3+ cells into the spinal cord and lung tissues. Moreover, our data revealed that EAE mice showed signs of respiratory disease, marked by increased vascular permeability, leukocyte influx, production of TNF-α and IL-17, perivascular and peribronchial inflammation, with pulmonary mechanical dysfunction associated with loss of respiratory volumes and elasticity, which DMF-encapsulated reverted in SLN nebulization. Conclusion: Our study suggests that inhalation of DMF-encapsulated SLN is an effective therapeutic protocol that reduces not only the CNS inflammatory process and disability progression, characteristic of EAE disease, but also protects mice from lung inflammation and pulmonary dysfunction. Neuroscience Group Department of Physiology and Biophysics Institute of Biological Sciences Federal University of Minas Gerais (UFMG), MG Nanoneurobiophysics Research Group Department of Physics Chemistry and Mathematics Federal University of Sao Carlos (UFSCAR), Sao Paulo State of Sao Paulo University (UNESP) Drugs and Medicines Department School of Pharmaceutical Sciences, Sao Paulo Laboratory of Pulmonary Immunology and Mechanics Department of Physiology and Biophysics Institute of Biological Sciences Federal University of Minas Gerais (UFMG), MG Laboratory of Immunology and Genomics of Parasites Department of Parasitology Institute of Biological Sciences Federal University of Minas Gerais (UFMG), MG Center for Technology and Research in Magneto-Resonance (CTPMAG) Federal University of Minas Gerais (UFMG), MG Department of Pathology Institute of Biological Sciences Federal University of Minas Gerais (UFMG), MG Laboratory of Functional Genetics Department of Genetics Ecology and Evolution Institute of Biological Sciences Federal University of Minas Gerais (UFMG), MG Department of Pharmacology Institute of Biological Sciences Federal University of Goias (UFG), GO State of Sao Paulo University (UNESP) Drugs and Medicines Department School of Pharmaceutical Sciences, Sao Paulo
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- 2021
31. Concomitant experimental coinfection by Plasmodium berghei NK65-NY and Ascaris suum downregulates the Ascaris-specific immune response and potentiates Ascaris-associated lung pathology
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Fabrício Marcus Silva Oliveira, Remo Castro Russo, Denise Silva Nogueira, Thaís Leal-Silva, Ricardo Toshio Fujiwara, Luiza de Lima Silva Padrão, Marcelo Vidigal Caliari, Lilian Lacerda Bueno, Lucas Kraemer, Ana Cristina Loiola Ruas, and Flaviane Vieira-Santos
- Subjects
Male ,0301 basic medicine ,Lung inflammation ,Plasmodium berghei ,Plasmodium berghei NK65-NY ,RC955-962 ,030231 tropical medicine ,Down-Regulation ,Infectious and parasitic diseases ,RC109-216 ,Lung injury ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Arctic medicine. Tropical medicine ,parasitic diseases ,medicine ,Animals ,Respiratory function ,Helminth infection ,Lung ,Ascaris suum ,Ascariasis ,medicine.diagnostic_test ,biology ,Coinfection ,business.industry ,Ascaris ,Research ,Pulmonary mechanics ,biology.organism_classification ,medicine.disease ,Immunity, Innate ,Malaria ,Mice, Inbred C57BL ,030104 developmental biology ,Infectious Diseases ,Bronchoalveolar lavage ,Gene Expression Regulation ,Immunology ,Parasitology ,business - Abstract
Background Ascariasis and malaria are highly prevalent parasitic diseases in tropical regions and often have overlapping endemic areas, contributing to high morbidity and mortality rates in areas with poor sanitary conditions. Several studies have previously aimed to correlate the effects of Ascaris-Plasmodium coinfections but have obtained contradictory and inconclusive results. Therefore, the present study aimed to investigate parasitological and immunopathological aspects of the lung during murine experimental concomitant coinfection by Plasmodium berghei and Ascaris suum during larvae ascariasis. Methods C57BL/6J mice were inoculated with 1 × 104P. berghei strain NK65-NY-infected red blood cells (iRBCs) intraperitoneally and/or 2500 embryonated eggs of A. suum by oral gavage. P. berghei parasitaemia, morbidity and the survival rate were assessed. On the seventh day postinfection (dpi), A. suum lung burden analysis; bronchoalveolar lavage (BAL); histopathology; NAG, MPO and EPO activity measurements; haematological analysis; and respiratory mechanics analysis were performed. The concentrations of interleukin (IL)-1β, IL-12/IL-23p40, IL-6, IL-4, IL-33, IL-13, IL-5, IL-10, IL-17A, IFN-γ, TNF and TGF-β were assayed by sandwich ELISA. Results Animals coinfected with P. berghei and A. suum show decreased production of type 1, 2, and 17 and regulatory cytokines; low leukocyte recruitment in the tissue; increased cellularity in the circulation; and low levels of NAG, MPO and EPO activity that lead to an increase in larvae migration, as shown by the decrease in larvae recovered in the lung parenchyma and increase in larvae recovered in the airway. This situation leads to severe airway haemorrhage and, consequently, an impairment respiratory function that leads to high morbidity and early mortality. Conclusions This study demonstrates that the Ascaris-Plasmodium interaction is harmful to the host and suggests that this coinfection may potentiate Ascaris-associated pathology by dampening the Ascaris-specific immune response, resulting in the early death of affected animals.
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- 2021
32. Nitric oxide contributes to liver inflammation and parasitic burden control in Ascaris suum infection
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Fabrício Marcus Silva Oliveira, Lucas Kraemer, Caroline Cavalcanti da Silva, Denise Silva Nogueira, Ana Clara Gazzinelli-Guimarães, Pedro Henrique Gazzinelli-Guimarães, Fernando Sérgio Barbosa, Nathalia Maria Resende, Marcelo Vidigal Caliari, Soraya Torres Gaze, Daniella Castanheira Bartholomeu, Ricardo Toshio Fujiwara, and Lilian Lacerda Bueno
- Subjects
Inflammation ,Ascariasis ,Immunology ,General Medicine ,Nitric Oxide ,Mice, Inbred C57BL ,Mice ,Infectious Diseases ,Liver ,Animals ,Cytokines ,Parasites ,Parasitology ,Ascaris suum - Abstract
Human ascariasis is one of the most prevalent neglected tropical diseases worldwide. The immune response during human ascariasis is characterized by Th2 polarization and a mixed Th2/Th17 response during the pathogenesis of experimental larval ascariasis. Cytokines and other pro-inflammatory mediators, such as nitric oxide (NO), are involved in helminthic infections. However, the role of NO in ascariasis remains unclear.Given the importance of NO in inflammation, we aimed to determine the immunological and histopathological alterations in the livers of C57BL/6 iNOSIn this study, parasitic load was evaluated in the livers of wild type C57BL/6 and C57BL/6 iNOSThe results showed that NO is important for controlling parasitic load during infection by A. suum. C57BL/6iNOSWe demonstrated that NO is a crucial inflammatory molecule during Ascaris sp. infection and controls the establishment of the parasite and the development of the host immune response in the liver.
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- 2022
33. Enhanced apoptotic index, chemokines and inflammatory recruitment in renal tissues shows relationship with the clinical signs in Leishmania-infected dogs
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Maria Norma Melo, Daniel Menezes-Souza, Luísa Mourão Dias Magalhães, Barbara Laurice Araújo Verçosa, Maria Imaculada Muniz-Junqueira, Anilton Cesar Vasconcelos, and Ricardo Toshio Fujiwara
- Subjects
Chemokine ,Inflammation ,CCL4 ,Apoptosis ,Kidney ,CCL5 ,Dogs ,medicine ,Animals ,Dog Diseases ,Leishmania infantum ,Caspase ,General Veterinary ,biology ,General Medicine ,medicine.anatomical_structure ,Real-time polymerase chain reaction ,Immunology ,biology.protein ,Leishmaniasis, Visceral ,Parasitology ,medicine.symptom ,Chemokines - Abstract
Apoptosis is associated with resolution of inflammation. However, apoptosis may also occur in active inflammation, balancing inflammatory recruitment instead of a resolution event. To test that hypothesis, we measured apoptosis and chemokines expression, involved in recruitment of inflammatory cells. Clinical affected and subclinically infected dogs with canine leishmaniosis (CanL) and uninfected controls were assessed. Apoptosis in renal tissue (glomeruli, tubules, and inflammatory infiltrate) and cellularity in inflammatory foci were quantified. Messenger RNA of CCL5, CCL4, MCP-1, MCP-2, Caspase (Casp) 3, Casp 8, Casp 9, Bax, Bcl2 and Fas were quantified by qRT PCR. Clinical affected dogs showed more intense inflammation and higher cellularity in the inflammatory infiltrates than subclinically infected ones, which were higher than controls. Glomerular and tubular cells showed higher apoptotic index in clinical affected dogs when compared to controls. Apoptosis within the inflammatory infiltrates was higher in clinical affected dogs. Bax/Bcl2 ratio and CCL4 showed higher expression in kidney from clinical affected when compared to subclinically infected dogs. Casp 3/CCL4 ratio expression were higher in subclinically infected dogs than in the clinical affected group. Additionally, results suggest that Casp 3/CCL4 ratio is balancing towards an inflammatory recruitment and CCL4 and Bax/Bcl2 ratio expression is associated with active inflammation in clinical affected CanL. Data demonstrate that apoptosis was not always correlated with resolution of inflammation, when a morphometric and a molecular evaluation were performed concomitantly. In kidneys of Leishmania infected dogs, apoptosis and chemokines may be balancing inflammatory recruitment. In conclusion, Bax/Bcl2 ratio, chemokines, Casp 8, Casp 3 and Fas were associated with renal apoptosis, active inflammation and increased inflammatory recruitment observed in clinical affected animals, influencing the clinical presentation of leishmaniosis.
- Published
- 2021
34. Serodiagnosis of canine leishmaniasis using a novel recombinant chimeric protein constructed with distinct B-cell epitopes from antigenic Leishmania infantum proteins
- Author
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Amanda S. Machado, Miguel A. Chávez-Fumagalli, Ricardo Toshio Fujiwara, Bruna B. Fernandes, Daniela P. Lage, Livia A. Alves, Fernanda F. Ramos, Camila S. Freitas, Williane Fernanda Siqueira, João A. Oliveira-da-Silva, Danniele L. Vale, Danielle F. de Magalhães-Soares, Thaís T.O. Santos, Nathália C. Galvani, Gabriel Paulino Luiz, Ricardo Andrez Machado-de-Ávila, Daysiane de Oliveira, Julia A.G. Silveira, Vívian T. Martins, Eduardo A.F. Coelho, Jamil S. Oliveira, and Lilian Lacerda Bueno
- Subjects
Recombinant Fusion Proteins ,Context (language use) ,Antigens, Protozoan ,Enzyme-Linked Immunosorbent Assay ,Sensitivity and Specificity ,Epitope ,law.invention ,Dogs ,Antigen ,law ,Canine leishmaniasis ,medicine ,Animals ,Serologic Tests ,Dog Diseases ,Leishmania infantum ,General Veterinary ,biology ,General Medicine ,medicine.disease ,biology.organism_classification ,Leishmania ,Virology ,Recombinant Proteins ,Visceral leishmaniasis ,Recombinant DNA ,Epitopes, B-Lymphocyte ,Leishmaniasis, Visceral ,Parasitology - Abstract
Visceral leishmaniasis (VL) is an important public health problem in the world, and control measures are insufficient to avoid the spread of this neglected disease. Dogs are important domestic reservoirs of Leishmania parasites in countries where VL is a zoonosis, representing a major source of infection between sand fly vectors and humans. In this context, a precise diagnosis of canine leishmaniasis (CanL) could help to reduce the number of human cases. Distinct approaches for the diagnosis of CanL have used recombinant proteins in serological assays. However, variable results of the antigens have been found, mainly to diagnosis asymptomatic cases. The present study used bioinformatics to select specific B-cell epitopes of four Leishmania infantum proteins, which had previously been proven to be antigenic in VL, aiming to produce a novel chimeric protein and to evaluate it for the diagnosis of CanL. Seven B-cell epitopes were identified and used to construct the chimera, which was analyzed in a recombinant format through an ELISA assay against a canine serological panel. A soluble Leishmania antigenic extract (SLA) was used as an antigen control. Results showed 100 % sensitivity and specificity for chimera, while when using SLA the values were 26.0 % and 96.4 %, respectively. The performance of chimera was compared with a commercial kit using asymptomatic and symptomatic dog sera, and the data showed that no false-negative result was found when the recombinant protein was used. However, when using the commercial kit, 40.0 % and 16.0 % of the false-negative results were found, respectively. In conclusion, the recombinant chimera showed an antigenic potential to be evaluated in new studies against a larger serological panel for the diagnosis of CanL.
- Published
- 2021
35. A recombinant protein (MyxoTLm) for the serological diagnosis of acute and chronic Trypanosoma vivax infection in cattle
- Author
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Ana Luiza Teixeira Silva, Álvaro Ferreira-Júnior, Mário Steindel, Edmundo C. Grisard, Lorena Lopes Ferreira, Mariana Santos Cardoso, Lilian Lacerda Bueno, Ricardo Toshio Fujiwara, Renato L. Santos, Daniella Castanheira Bartholomeu, Luiz Claudio Miletti, and Guilherme Rafael Gomide Pinheiro
- Subjects
0301 basic medicine ,Anemia ,030231 tropical medicine ,Cattle Diseases ,Antigens, Protozoan ,Enzyme-Linked Immunosorbent Assay ,Parasitemia ,Asymptomatic ,Serology ,03 medical and health sciences ,Lethargy ,0302 clinical medicine ,Antigen ,medicine ,Animals ,Trypanosoma vivax ,General Veterinary ,biology ,Trypanosomiasis, Bovine ,General Medicine ,030108 mycology & parasitology ,medicine.disease ,biology.organism_classification ,Recombinant Proteins ,Immunology ,Parasitology ,Cattle ,Lymph ,medicine.symptom - Abstract
Human trypanosomiases and animal trypanosomoses are caused by distinct protozoan parasites of the genus Trypanosoma. The etiological agents of bovine trypanosomosis (BT) are T. vivax, T. congolense, or T. brucei, whose acute infections are initially characterized by hyperthermia, following moderate to severe anemia, subcutaneous edema, lethargy, reduced milk production, progressive weight loss, enlarged lymph nodes, reproductive disorders and death. Animals that survive the acute phase might recover and progress to the chronic, often asymptomatic, phase of infection. Despite their low sensitivity due to the characteristic low parasitemia, simple and costless direct parasitological examinations are the preferred diagnostic methods for animals. Thus, most of the epidemiological studies of BT are based on serological techniques using crude antigen. In this study, we describe the use of the MyxoTLm recombinant protein as an antigen on serological assays. Anti-T. vivax IgM and anti-T. vivax IgG ELISA assays using purified MyxoTLm revealed specificity rates of 91.30 % and 95.65 % and sensitivity rates of 82.35 % and 88.23 %, respectively, being higher than reported for crude antigens. Also, MyxoTLm demonstrated a good performance to detect IgM (ROC curve area = 0.8568) and excellent performance to detect IgG (ROC curve area = 0.9565) when compared to a crude antigen. T. evansi crude antigen used in the indirect anti-T. vivax IgM ELISA reached 70.58 % sensitivity and 78.26 % specificity, and had a lower test performance (ROC curve area = 0.7363). When applied to the anti-T. vivax IgG ELISA, the crude antigen reached 82.35 % sensitivity and 69.56 % specificity, also presenting a low performance with area under the ROC curve of 0.7570. Therefore, the use of MyxoTLm as an antigen on serological diagnosis of BT revealed to increase the sensitivity and the specificity if compared to crude antigens.
- Published
- 2021
36. Allergen presensitization drives an eosinophil-dependent arrest in lung-specific helminth development
- Author
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Thomas B. Nutman, Sandra Bonne-Année, Joshua Sciurba, Erik P. Karmele, Pedro Henrique Gazzinelli-Guimarães, Alessandra Ricciardi, Ricardo Toshio Fujiwara, and Rafael de Queiroz Prado
- Subjects
CD4-Positive T-Lymphocytes ,Male ,0301 basic medicine ,Swine ,Biology ,Allergic inflammation ,Type 2 immune response ,Allergic sensitization ,Mice ,03 medical and health sciences ,Th2 Cells ,0302 clinical medicine ,Hypersensitivity ,medicine ,Animals ,Antigens, Dermatophagoides ,Lung ,Sensitization ,Inflammation ,Antigen Presentation ,Ascariasis ,Mice, Inbred BALB C ,Innate immune system ,Ascaris ,Macrophages ,Pyroglyphidae ,Innate lymphoid cell ,General Medicine ,Allergens ,Eosinophil ,biology.organism_classification ,Asthma ,Immunity, Innate ,Eosinophils ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Immunology ,Female ,Research Article - Abstract
This study investigates the relationship between helminth infection and allergic sensitization by assessing the influence of preexisting allergy on the outcome of helminth infections, rather than the more traditional approach in which the helminth infection precedes the onset of allergy. Here we used a murine model of house dust mite–induced (HDM-induced) allergic inflammation followed by Ascaris infection to demonstrate that allergic sensitization drives an eosinophil-rich pulmonary type 2 immune response (Th2 cells, M2 macrophages, type 2 innate lymphoid cells, IL-33, IL-4, IL-13, and mucus) that directly hinders larval development and reduces markedly the parasite burden in the lungs. This effect is dependent on the presence of eosinophils, as eosinophil-deficient mice were unable to limit parasite development or numbers. In vivo administration of neutralizing antibodies against CD4 prior to HDM sensitization significantly reduced eosinophils in the lungs, resulting in the reversal of the HDM-induced Ascaris larval killing. Our data suggest that HDM allergic sensitization drives a response that mimics a primary Ascaris infection, such that CD4(+) Th2-mediated eosinophil-dependent helminth larval killing in the lung tissue occurs. This study provides insight into the mechanisms underlying tissue-specific responses that drive a protective response against the early stages of the helminths prior to their establishing long-lasting infections in the host.
- Published
- 2019
37. In vitro activity evaluation of seven Brazilian Asteraceae against cancer cells and Leishmania amazonensis
- Author
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Ricardo Toshio Fujiwara, Ana Cláudia Nogueira da Silva, Raquel Martins de Almeida, Daisy Jereissati Barbosa Lima, Manoel Odorico de Moraes Filho, A. M. Nascimento, Débora Caroline do Nascimento Rodrigues, Sebastião Rodrigo Ferreira, Paulo Michel Pinheiro Ferreira, Renato Malveira Carreiro do Nascimento, and Cláudia Pessoa
- Subjects
0106 biological sciences ,Traditional medicine ,biology ,Plant Science ,Asteraceae ,biology.organism_classification ,01 natural sciences ,Terpenoid ,In vitro ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,chemistry.chemical_compound ,Phytochemical ,chemistry ,Cell culture ,Growth inhibition ,Amastigote ,010606 plant biology & botany ,Lupeol - Abstract
Natural products are a very productive leading source of compounds for the development of medicines. The aim of this research was to investigate the in vitro antiproliferative and antileishmanial activities of 21 extracts from 7 Asteraceae species. Dried aerial parts of Asteraceae plant species were extracted using organic solvents in order of increasing polarities. Antiproliferative activity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method, and nine extracts (43%) displayed moderate to high in vitro antiproliferative activity against human cancer cell lines HCT-116, OVCAR-8 and SF-295. Two crude extracts displaying antiproliferative activity were fractionated and yielded a fraction rich in lupeol acetate, a fraction rich in lupeol and 5,7,3′,4′-tetrahydroxyflavone-7-O-β-D-glucopyranoside. The antileishmanial activity was evaluated by amastigotes growth inhibition test in Leishmania amazonensis, and three extracts showed promising activity. Phytochemical screening of all plant extracts was also made for terpenoids, flavonoids, tannins, alkaloids and saponins.
- Published
- 2019
38. Unraveling Ascaris suum experimental infection in humans
- Author
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Fernando Sérgio Barbosa, Ricardo Toshio Fujiwara, Ana Clara Gazzinelli-Guimarães, Agostinho Gonçalves Viana, Anderson C. dos Santos, Daniella Castanheira Bartholomeu, Pedro Henrique Gazzinelli-Guimarães, Luísa Mourão Dias Magalhães, Thaís Eloi da Silva, Nathalia Maria Resende, Lilian Lacerda Bueno, Denise Silva Nogueira, Chiara Cássia Oliveira Amorim, and Stefan M. Geiger
- Subjects
0301 basic medicine ,Swine ,medicine.medical_treatment ,030106 microbiology ,Immunology ,Microbiology ,03 medical and health sciences ,Immune system ,Ascariasis ,parasitic diseases ,medicine ,Animals ,Humans ,Respiratory system ,Ascaris suum ,Swine Diseases ,biology ,respiratory system ,biology.organism_classification ,medicine.disease ,Diarrhea ,030104 developmental biology ,Infectious Diseases ,Cytokine ,Larva ,biology.protein ,medicine.symptom ,Antibody ,Ascaris lumbricoides - Abstract
Ascaris lumbricoides and Ascaris suum are two closely related parasites that infect humans and pigs. The zoonotic potential of A. suum has been a matter of debate for decades. Here we sought to investigate the potential human infection by A. suum and its immunological alterations. We orally infected five healthy human subjects with eggs embraced by A. suum. The infection was monitored for symptoms and possible respiratory changes, by an interdisciplinary health team. Parasitological, hematological analyses, serum immunoglobulin, cytokine profiles, and gene expression were evaluated during the infection. Our results show that A. suum is able to infect and complete the cycle in humans causing A. lumbricoides similar symptoms, including, cough, headache, diarrhea, respiratory discomfort and chest x-ray alterations coinciding with larvae migration in the lungs. We also observed activation of the immune system with production of IgM and IgG and a Th2/Th17 response with downregulation of genes related to Th1 and apoptosis. PCA (Principal componts analysis) show that infection with A. suum leads to a change in the immune landscape of the human host. Our data reinforce the zoonotic capacity of A. suum and bring a new perspective on the understanding of the immune response against this parasite.
- Published
- 2021
39. Investigação sorológica da Leishmaniose Felina em abrigo de animais de área endêmica para a doença no nordeste do Brasil utilizando a proteína rKDDR-plus
- Author
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Ricardo Toshio Fujiwara, Victor Fernando Santana Lima, Tatyane Martins Cirilo, Ana Carolina Amado Gomes, Sofia Cerqueira Schettino, Matheus Resende Oliveira, Luiz Fernando de Jesus Nascimento, Silvio Santana Dolabella, and Wemerson de Santana Neres
- Published
- 2021
40. Effect of Triatoma Infestans Saliva on Mouse Immune System Cells: The Role of the Pore-Forming Salivary Protein Trialysin
- Author
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Fernanda Faria Rocha, Pedro H. Gazzinelli-Guimarães, Adriana Coelho Soares, Rodrigo A. Lourdes, Ligia R. Estevao, Milene Alvarenga Rachid, Lilian Lacerda Bueno, Nelder Figueiredo Gontijo, Marcos Horacio Pereira, Mauricio Viana Sant'Anna, Ulisses Antonio Natividade, Ricardo Toshio Fujiwara, and Ricardo Nascimento Araujo
- Subjects
History ,Polymers and Plastics ,Business and International Management ,Industrial and Manufacturing Engineering - Published
- 2021
41. Phenotypic, functional and serological aspects of genotypic-specific immune response of experimental T. cruzi infection
- Author
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Ricardo Toshio Fujiwara, Olindo Assis Martins-Filho, João Luís Reis-Cunha, Tiago Antônio de Oliveira Mendes, Marta de Lana, Maurício Azevedo Batista, Glaucia Diniz Alessio, Daniella Castanheira Bartholomeu, Patrick Orestes de Azevedo, Rodrigo de Almeida Lourdes, and Denise da Silveira-Lemos
- Subjects
0301 basic medicine ,Chagas disease ,Genotype ,Veterinary (miscellaneous) ,Trypanosoma cruzi ,030231 tropical medicine ,Context (language use) ,Parasitemia ,Biology ,Serology ,Pathogenesis ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Immune system ,Antigen ,parasitic diseases ,medicine ,Animals ,Chagas Disease ,Immunity ,030108 mycology & parasitology ,medicine.disease ,Acquired immune system ,Mice, Inbred C57BL ,Infectious Diseases ,Phenotype ,Insect Science ,Immunology ,Parasitology - Abstract
The complexity and multifactorial characteristics of Chagas disease pathogenesis hampers the establishment of appropriate experimental/epidemiological sets, and therefore, still represents one of the most challenging fields for novel insights and discovery. In this context, we used a set of attributes including phenotypic, functional and serological markers of immune response as candidates to decode the genotype-specific immune response of experimental T. cruzi infection. In this investigation, we have characterized in C57BL/6 J mice, the early (parasitemia peak) and late (post-parasitemia peak) aspects of the immune response elicited by T. cruzi strains representative of TcI, TcII or TcVI. The results demonstrated earlier parasitemia peak for TcII/Y strain followed by TcVI/CL-Brener and TcI/Colombiana strains. A panoramic overview of phenotypic and functional features of the TCD4+, TCD8+ and B-cells from splenocytes demonstrated that mice infected with TcI/Colombiana strain exhibited at early stages of infection low levels of most cytokine+ cells with a slight increase at late stages of infection. Conversely, mice infected with TcII/Y strain presented an early massive increase of cytokine+ cells, which decreases at late stages. The TcVI/CL-Brener strain showed an intermediate profile at early stages of infection with a slight increase later on at post-peak of parasitemia. The panoramic analysis of immunological connectivity demonstrated that early after infection, the TcI/Colombiana strain trigger immunological network characterized by a small number of connectivity, selectively amongst cytokines that further shade towards the late stages of infection. In contrast, the TcII/Y strain elicited in more imbricate networks early after infection, comprising a robust number of interactions between pro-inflammatory mediators, regulatory cytokines and activation markers that also decrease at late infection. On the other hand, the infection with TcVI/CL-Brener strain demonstrated an intermediate profile with connectivity axes more stable at early and late stages of infection. The analysis of IgG2a reactivity to AMA, TRYPO and EPI antigens revealed that at early stages of infection, the genotype-specific reactivity to AMA, TRYPO and EPI to distinguish was higher for TcI/Colombiana as compared to TcII/Y and TcVI/CL while, at late stages of infection, higher reactivity to AMA was observed in mice infected with TcVI/CL and TcII/Y strains. The novel systems biology approaches and the use of a flow cytometry platform demonstrated that distinct T. cruzi genotypes influenced in the phenotypic and functional features of the host immune response and the genotype-specific serological reactivity during early and late stages of experimental T. cruzi infection.
- Published
- 2020
42. Diagnostic comparison of stool exam and point-of-care circulating cathodic antigen (POC-CCA) test for schistosomiasis mansoni diagnosis in a high endemicity area in northeastern Brazil
- Author
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Fernando Schemelzer de Moraes Bezerra, Luciene Barbosa, Marta Cristhiany Cunha Pinheiro, Agostinho Gonçalves Viana, Danielle de Freitas Bezerra, and Ricardo Toshio Fujiwara
- Subjects
Adult ,Male ,Veterinary medicine ,Adolescent ,Endemic Diseases ,030231 tropical medicine ,Schistosomiasis ,Enzyme-Linked Immunosorbent Assay ,Urine ,Biology ,03 medical and health sciences ,Feces ,Young Adult ,0302 clinical medicine ,Antigen ,parasitic diseases ,medicine ,Prevalence ,Animals ,Humans ,030212 general & internal medicine ,Child ,Eggs per gram ,Point of care ,Endemic area ,Schistosoma mansoni ,Middle Aged ,medicine.disease ,biology.organism_classification ,Schistosomiasis mansoni ,Test (assessment) ,Infectious Diseases ,ROC Curve ,Point-of-Care Testing ,Antigens, Helminth ,Child, Preschool ,Animal Science and Zoology ,Parasitology ,Female ,Brazil - Abstract
This study aimed to evaluate the performance of the point-of-care circulating cathodic antigen (POC-CCA) test in a highly endemic area in Brazil, comparing it to the Kato-Katz (KK) technique for sensitivity, specificity and the intensity of the reaction of the test in relation to the parasitic load. The community in Sergipe, Brazil, participated in the study, providing three stool samples, one of urine (POC-CCA) and fingers tick blood sample was tested by enzyme-linked immunosorbent assay (ELISA). Sensitivity, specificity, positive predictive value, negative predictive value, accuracy, kappa coefficient and Spearman's correlation were calculated for the POC-CCA test using the KK as the reference. The prevalence of schistosomiasis by KK testing was 48.82%; POC-CCA (t+) 66.14%; POC-CCA (t−) 45.24%. ELISA results showed 100% agreement in individuals with high and moderate eggs per gram (EPG). POC-CCA presented good diagnostic performance in individuals with medium and high EPG, but there were a high number of false negatives in individuals with low intensity infections. As observed, POC-CCA-filter test improves accuracy and sensitivity compared to a conventional test.
- Published
- 2020
43. Antileishmanial activity of fullerol and its liposomal formulation in experimental models of visceral leishmaniasis
- Author
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Yuri Machado, Marina Ladeira, Maurício B.V. Pinheiro, Virgínia M.R. Vallejos, Priscila G. Reis, Frédéric Frézard, Ricardo Toshio Fujiwara, Guilherme Santos Ramos, Daniel Menezes Souza, Maria Norma Melo, and Luiz Orlando Ladeira
- Subjects
0301 basic medicine ,Drug Compounding ,Leishmania mexicana ,RM1-950 ,Pharmacology ,Parasite load ,Parasite Load ,Immunomodulation ,03 medical and health sciences ,0302 clinical medicine ,In vivo ,medicine ,Animals ,Leishmania infantum ,Amastigote ,Leishmaniasis ,Liposome ,Mice, Inbred BALB C ,biology ,Mesocricetus ,business.industry ,General Medicine ,Fullerol ,medicine.disease ,Leishmania ,biology.organism_classification ,Fullerenol ,Lipids ,Trypanocidal Agents ,Disease Models, Animal ,030104 developmental biology ,Visceral leishmaniasis ,Liver ,030220 oncology & carcinogenesis ,Drug delivery ,Liposomes ,Macrophages, Peritoneal ,Cytokines ,Leishmaniasis, Visceral ,Nanoparticles ,Female ,Therapeutics. Pharmacology ,Fullerenes ,Inflammation Mediators ,business - Abstract
Visceral leishmaniasis (VL) is a systemic parasitic disease that leads to high rates of morbidity and mortality in humans worldwide. There is a great need to develop new drugs and novel strategies to make chemotherapy for this disease more efficacious and well tolerated. Recent reports on the immunomodulatory effects and the low toxicity of the spherical carbon nanostructure fullerol led us to investigate in vitro and in vivo antileishmanial activity in free and encapsulated forms in liposomes. When assayed against intramacrophagic Leishmania amastigotes, fullerol showed a dose-dependent reduction of the infection index with IC50 of 0.042 mg/mL. When given daily by i.p. route for 20 days (0.05 mg/kg/d) in a murine model of acute VL, fullerol promoted significant reduction in the liver parasite load. To improve the delivery of fullerol to the infection sites, liposomal formulations were prepared by the dehydration-rehydration method. When evaluated in the acute VL model, liposomal fullerol (Lip-Ful) formulations given i.p. at 0.05 and 0.2 mg/kg with 4-days intervals were more effective than the free form, with significant parasite reductions in both liver and spleen. Lip-Ful at 0.2 mg/kg promoted complete parasite elimination in the liver. The antileishmanial activity of Lip-Ful was further confirmed in a chronic model of VL. Lip-Ful was also found to induce secretion of pro-inflammatory TNF-α, IFN-γ and IL-1β cytokines. In conclusion, this work reports for the first time the antileishmanial activity of fullerol and introduces an innovative approach for treatment of VL based on the association of this nanostructure with liposomes.
- Published
- 2020
44. Ketamine can be produced by Pochonia chlamydosporia: an old molecule and a new anthelmintic?
- Author
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Valentina N. Maciel, Alan Rodrigues Teixeira Machado, Rodrigo Maia de Pádua, Fernando Sérgio Barbosa, Ricardo Toshio Fujiwara, Jackson Victor de Araújo, Sebastião Rodrigo Ferreira, Raquel Martins de Almeida, Luis Fernando Furtado, Lorendane Millena de Carvalho, Thiago de Oliveira Mendes, Élida Mara Leite Rabelo, Lúcia Pinheiro Santos Pimenta, Daniella Castanheira Bartholomeu, Lilian Lacerda Bueno, and José Hugo de Sousa Gomes
- Subjects
0301 basic medicine ,Ancylostoma ,Nematoda ,030106 microbiology ,Albendazole ,Nematicidal molecule ,Nematophagous fungus ,Microbiology ,lcsh:Infectious and parasitic diseases ,03 medical and health sciences ,Mice ,Ivermectin ,Pochonia chlamydosporia ,Cricetinae ,parasitic diseases ,medicine ,Animals ,lcsh:RC109-216 ,Anthelmintic ,Caenorhabditis elegans ,Pest Control, Biological ,Ascaris suum ,Ancylostoma ceylanicum ,biology ,Research ,Antinematodal Agents ,biology.organism_classification ,New drugs ,030104 developmental biology ,Infectious Diseases ,Nematode ,Parasitology ,Hypocreales ,Ketamine ,medicine.drug - Abstract
Background Infection by nematodes is a problem for human health, livestock, and agriculture, as it causes deficits in host health, increases production costs, and incurs a reduced food supply. The control of these parasites is usually done using anthelmintics, which, in most cases, have not been fully effective. Therefore, the search for new molecules with anthelmintic potential is necessary. Methods In the present study, we isolated and characterized molecules from the nematophagous fungus Pochonia chlamydosporia and tested these compounds on three nematodes: Caenorhabditis elegans; Ancylostoma ceylanicum; and Ascaris suum. Results The ethyl acetate extract showed nematicidal activity on the nematode model C. elegans. We identified the major substance present in two sub-fractions of this extract as ketamine. Then, we tested this compound on C. elegans and the parasites A. ceylanicum and A. suum using hamsters and mice as hosts, respectively. We did not find a difference between the animal groups when considering the number of worms recovered from the intestines of animals treated with ketamine (6 mg) and albendazole (P > 0.05). The parasite burden of larvae recovered from the lungs of mice treated with ketamine was similar to those treated with ivermectin. Conclusions The results presented here demonstrate the nematicidal activity of ketamine in vitro and in vivo, thus confirming the nematicidal potential of the molecule present in the fungus P. chlamydosporia may consist of a new method of controlling parasites.
- Published
- 2020
45. The balance between IL-12/IL4 in renal tissue switches the inflammatory response arm and shows relationship with the clinical signs in Leishmania-infected dogs
- Author
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Maria Imaculada Muniz-Junqueira, Anilton Cesar Vasconcelos, Ricardo Toshio Fujiwara, Maria Norma Melo, Daniel Menezes-Souza, and Barbara Laurice Araújo Verçosa
- Subjects
Male ,Pathology ,medicine.medical_specialty ,040301 veterinary sciences ,Immunology ,H&E stain ,Inflammation ,Biology ,Kidney ,Asymptomatic ,0403 veterinary science ,Pathogenesis ,03 medical and health sciences ,Dogs ,medicine ,Animals ,Interferon gamma ,Dog Diseases ,Leishmania infantum ,Leishmaniasis ,030304 developmental biology ,0303 health sciences ,General Veterinary ,Interleukin ,04 agricultural and veterinary sciences ,Interleukin-12 ,medicine.anatomical_structure ,Gene Expression Regulation ,Tumor necrosis factor alpha ,Female ,Interleukin-4 ,medicine.symptom ,medicine.drug - Abstract
The pathogenesis of Canine leishmaniosis (CanL) is associated with altered cytokine expression and parasitic tissue shows a lot of inflammation. The aim of this study was to assess the renal inflammation and cytokine expression in eight symptomatic and eight asymptomatic Leishmania- infected dogs, and seven uninfected control dogs. Kidney fragments were stained with hematoxylin and eosin for morphometric evaluation. mRNA expression levels of interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), interleukin (IL)-2, IL-4, IL-10, and IL-12 were assessed in the kidney fragments using quantitative real time-polymerase chain reaction. Inflammation, quantified by the average area of the infiltrated immune cells, was greater in symptomatic dogs than in those asymptomatic, whereas asymptomatic dogs exhibited higher inflammation than the control dogs (p0.05, Tukey's test). Expression levels of IFN-γ, TNF-α, IL-4, IL-10, and IL-12 were upregulated in symptomatic dogs and downregulated in asymptomatic dogs compared with those of the uninfected group. Furthermore, IL-4 showed higher expression in symptomatic dogs than in asymptomatic ones (p0.05, Mann-Whitney test), which was directly associated with clinical manifestations (p0.05, Chi-square test). However, IL-12 was predominantly expressed in symptomatic dogs, shifting the balance from IL-12/IL-4 to IL-12, which elicits a change in the inflammatory response. Leishmania was not found in the renal tissues in any one of the studied groups. Our data suggests that the balance between IL-12 and IL-4 plays an important role in the regulation of inflammation in renal tissue and clinical presentations in CanL.
- Published
- 2020
46. Ketamine can be produced by Pochonia chlamydosporia: an old molecule and a new role
- Author
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Sebastiao Rodrigo Ferreira, Alan Rodrigues T. Machado, Luís Fernando Furtado, Jose Hugo de S. Gomes, Raquel M. de Almeida, Thiago de Oliveira Mendes, Valentina N. Maciel, Fernando Sergio Barbosa, Lorendane M. Carvalho, Lilian Lacerda Bueno, Daniella Castanheira Bartholomeu, Jackson Victor de Araújo, Elida M. L. Rabelo, Rodrigo Maia de Pádua, Lucia Pinheiro Santos Pimenta, and Ricardo Toshio Fujiwara
- Abstract
Background: Infection by nematodes is a problem for human health, livestock, and agriculture, as it causes deficits in host health, increases production costs, and incurs less food supply. The control of these parasites is usually done using anthelmintics, which, in most cases, have not been fully effective. Therefore, the search for new molecules with anthelmintic potential is necessary. Methods: In the present study, we isolated and characterized molecules from the nematophagous fungus Pochonia chlamydosporia and tested these compounds on three nematodes: Caenorhabditis elegans, Ancylostoma ceylanicum, and Ascaris suum. Results: The ethyl acetate extract showed nematicidal activity on the nematode model C. elegans. We identified the major substance present in two subfractions of this extract as ketamine. Then, we tested this compound on C. elegans and the parasites A. ceylanicum and A. suum using hamsters and mice as hosts, respectively. We did not find a difference between the animal groups when considering the number of worms recovered from the intestines of animals treated with ketamine and albendazole (P ˂ 0.05). The parasite burden of larvae recovered from the lungs of mice treated with ketamine was similar to those treated with ivermectin. Conclusion: These results presented here demonstrate the nematicidal activity of ketamine in vitro and in vivo. Thus, confirming the nematicidal potential of the molecule present in the fungus P. chlamydosporia may consist a new method of controlling parasites.
- Published
- 2020
47. Immunopathology and modulation induced by hookworms: From understanding to intervention
- Author
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Lívia Silva Araújo Passos, Ricardo Toshio Fujiwara, Luísa Mourão Dias Magalhães, and Lilian Lacerda Bueno
- Subjects
0301 basic medicine ,Ancylostomatoidea ,Male ,030231 tropical medicine ,Immunology ,Antibodies, Helminth ,Host-Parasite Interactions ,Immunomodulation ,03 medical and health sciences ,Hookworm Infections ,Soil ,0302 clinical medicine ,Intervention (counseling) ,Immunopathology ,parasitic diseases ,Excretory secretory antigens ,Prevalence ,Animals ,Humans ,Parasite transmission ,Hookworm infection ,Immunity, Cellular ,Vaccines ,biology ,biology.organism_classification ,Vaccination ,030104 developmental biology ,Neglected tropical diseases ,Parasitology - Abstract
Hookworm infection is considered the most prevalent human soil-transmitted helminth infection affecting approximately 500 million people and accounting for 3.2 million disability-adjusted life years lost annually. As with many other neglected tropical diseases, no international surveillance mechanisms that show accurate data on the prevalence of hookworm infection are in place, thus hindering strategies to control parasite transmission. In this review, we unravel the current knowledge in immunopathology and immunoregulation of hookworm infection, and present discoveries in drug therapies based on the capability of hookworms to regulate inflammation to treat allergic, inflammatory, and metabolic diseases. Additionally, we highlight potential vaccine development and treatments, and propose avenues for further inquiry.
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- 2020
48. Time-course of changes in performance, biomechanical, physiological and perceptual responses following resistance training sessions
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Karine Naves de Oliveira Goulart, Leszek Antoni Szmuchrowski, Bruno Pena Couto, Luciana Maria Oliveira, Marcos Daniel Motta Drummond, Nathalia Maria Resende, Ricardo Toshio Fujiwara, and Fernando Vitor Lima
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Adult ,Male ,medicine.medical_specialty ,Time Factors ,genetic structures ,media_common.quotation_subject ,Physical Exertion ,030209 endocrinology & metabolism ,Physical Therapy, Sports Therapy and Rehabilitation ,Audiology ,Athletic Performance ,03 medical and health sciences ,Young Adult ,0302 clinical medicine ,Perception ,Isometric Contraction ,medicine ,Humans ,Orthopedics and Sports Medicine ,skin and connective tissue diseases ,Association (psychology) ,Creatine Kinase ,media_common ,Resistance training ,Resistance Training ,030229 sport sciences ,General Medicine ,Myalgia ,Biomechanical Phenomena ,Volume load ,Time course ,Muscle Fatigue ,Exercise Test ,sense organs ,Percept ,Psychology - Abstract
This study determined the time-course of recovery after resistance training (RT) sessions and the association between changes in performance with changes in biomechanical, physiological and perceptual parameters. After a 4-week familiarization period, 14 resistance-trained males performed 3 experimental conditions, each one including 2 sessions with a recovery interval of 24, 48 h or 72 h, in a randomized order. RT sessions consisted of 5 sets of 8-10RM on squat and leg press exercises. The resistance was equal for the 2 sessions of each condition and repetitions were performed until concentric failure. Volume load (VL) and first set volume load (FSVL) were compared between sessions. Tests before each session included countermovement jump (CMJ), maximal voluntary isometric contraction (MVIC), creatine kinase (CK) and delayed onset muscle soreness (DOMS). (2 × 3) ANOVA with effect sizes (ES) assessed the time-course of recovery and Kendall test the correlation between variables (
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- 2020
49. A multiplex PCR protocol for rapid differential identification of four families of trematodes with medical and veterinary importance transmitted by Biomphalaria Preston, 1910 snails
- Author
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Ricardo Toshio Fujiwara, João Luís Reis-Cunha, Mariana Santos Cardoso, Daniella Castanheira Bartholomeu, Gabriela F. Rodrigues-Luiz, Hudson Alves Pinto, Silvia Gonçalves Mesquita, Roberta Lima Caldeira, Lilian Lacerda Bueno, and Cristiane Lafetá Furtado De Mendonça
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0301 basic medicine ,Veterinary medicine ,Veterinary (miscellaneous) ,030231 tropical medicine ,Biomphalaria ,Trematode Infections ,law.invention ,Host-Parasite Interactions ,03 medical and health sciences ,0302 clinical medicine ,Schistosomatidae ,law ,parasitic diseases ,Multiplex polymerase chain reaction ,medicine ,Parasite hosting ,Animals ,Humans ,Cercaria ,Polymerase chain reaction ,Disease Reservoirs ,biology ,030108 mycology & parasitology ,South America ,biology.organism_classification ,medicine.disease ,Infectious Diseases ,Insect Science ,Larva ,Coinfection ,Parasitology ,Schistosoma mansoni ,Trematoda ,Multiplex Polymerase Chain Reaction ,Specific identification - Abstract
Trematodes have complex life cycles with multiple hosts. Biomphalaria snails commonly act as the first intermediate hosts of several species that can affect human and animal health. The specific identification of larval trematodes found in snails is difficult and limited, since the taxonomy of these flukes is based on morphological traits of the adults found in vertebrates. Despite recent advances worldwide, studies aiming at the use of molecular tools for the identification of cercariae found in snails are scarce in the South America. In fact, most studies are focused on Schistosoma mansoni, with few efforts directed towards the identification of larvae of other parasites found in planorbids. When reported, these other parasites are identified as cercarial types, an artificial morphological system of classification. Therefore, alternative strategies for a correct, rapid and inexpensive identification of larval trematodes found in Biomphalaria are needed. This work aimed at developing a methodology capable of distinguishing four important families of trematodes (Clinostomidae, Echinostomatidae, Schistosomatidae and Strigeidae) commonly found infecting species of Biomphalaria. Using the rDNA sequences of 34 species as input for the online tool TipMT, we designed trematode family-specific primers targeting the ITS region optimized to be used in multiplex PCR. The panel of primers identified in this study was effective at the same PCR condition. The specificity of the primers was confirmed, and the PCR sensitivity ranged from 0.1 ng to 1 ag of the DNA of the parasite. This methodology was also effective for the detection of coinfection. Through a simple, fast, accurate, and inexpensive methodology, it is possible to properly identify the trematode families included in this study in a single PCR reaction. A family level identification provides important information about probable hosts, pattern of life cycle and possible impacts that the infection generates in a specific region, thus allowing the design of better control strategies, especially for those infections that have medical and veterinary importance.
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- 2020
50. Application of Poloxamers for the Development of Drug Delivery System to Treat Leishmaniasis: A Review
- Author
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Silvio Santana Dolabella, Rogéria De Souza Nunes, Sona Jain, Audrey Rouse Soares Tavares Silva, Ricardo Scher, Amanda M.B. Costa, Eduardo A.F. Coelho, and Ricardo Toshio Fujiwara
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Pharmacology ,medicine.medical_specialty ,business.industry ,Clinical Biochemistry ,Antiprotozoal Agents ,Tropical disease ,Context (language use) ,Leishmaniasis ,Poloxamer ,medicine.disease ,Drug Delivery Systems ,Disease severity ,Drug Discovery ,Drug delivery ,Molecular Medicine ,Medicine ,Humans ,Nanotechnology ,business ,Intensive care medicine - Abstract
Leishmaniasis is a neglected tropical disease affecting more than 1.5 million people annually, with an annual mortality of over 20.000. The drugs used for its treatment are toxic, expensive, require extended treatment times and present variable efficacy. The disease severity and therapy limitations suggest the need for new antileishmanial agents. In this context, in order to identify new options for treatment, a number of studies based on nanotechnological strategies have been carried out. Poloxamers are triblock copolymers very often utilized for nanotherapeutic solutions, resulting in products with better solubility, higher stability, superior therapeutic efficacy and less toxicity. This review will discuss the physicochemical properties of the copolymers, as well as describe the use of poloxamers for the development of therapeutic formulations to treat leishmaniasis.
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- 2020
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