Carga parasitária, iNOS, citocinas e histopatologia na infecção por Leishmania infantum em cães com apresentações clínicas distintas

Submitted by Janaína Nascimento ([email protected]) on 2019-12-16T14:46:51Z No. of bitstreams: 1 ve_Vasconcelos_Tassia_etal_INI_2019.pdf: 2886512 bytes, checksum: 4b19381817d25d0019c92a2184f6aec5 (MD5)Approved for entry into archive by Janaína Nascimento ([email protected]) on 2019-12-16T15:01:48Z (GMT) No. of bitstreams: 1 ve_Vasconcelos_Tassia_etal_INI_2019.pdf: 2886512 bytes, checksum: 4b19381817d25d0019c92a2184f6aec5 (MD5)Made available in DSpace on 2019-12-16T15:01:48Z (GMT). No. of bitstreams: 1 ve_Vasconcelos_Tassia_etal_INI_2019.pdf: 2886512 bytes, checksum: 4b19381817d25d0019c92a2184f6aec5 (MD5) Previous issue date: 2019Universidade Federal Fluminense. Programa de Pós-graduação em Medicina Veterinária. Niterói, RJ, Brasil.Universidade Federal Fluminense. Programa de Pós-graduação em Medicina Veterinária. Niterói, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunoparasitologia. Rio de Janeiro, RJ, Brasil.Universidade Federal de São João del-Rei. Divinópolis, MG, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Celular. Curitiba, PR, Brasil.A Leishmaniose visceral (LV) é uma doença zoonótica com reservatório canino na América do Sul. Cães oriundos de área endemica brasileira, naturalmente infectados por Leishmania infantum, apresentando doença clínica severa (CS) ou doença branda ou ausente (BA) foram avaliados. Carga parasitária, histopatologia e expressão de mRNA de citocinas e iNOS foram analisados em baço e fígado, buscando determinar possíveis marcadores de susceptibilidade ou resistência à doença. Como principais resultados, tanto cães CS como BA apresentaram alta carga parsitária. IFN-γ foi a citocina mais expressiva em ambos os órgãos, sendo IL-6 e IL-4 também detectadas em baço e fígado e IL-10 em fígado. No tecido hepático foram encontradas as maiores medianas de IFN-γ e IL-10, sendo o fígado o principal órgão produtor de citocinas, com IL-10 sugerindo acompanhamento regulatório. Granulomas foram detectados em ambos os órgãos. Quando de sua ausência no baço, essa foi associada à elevação dos níveis de IL-6, salientando o papel anti-inflamatório dessa citocina. Alterações microscópicas foram principalmente caracterizadas por extensiva desorganização de polpa branca, com a resposta esplênica sendo sugerida como subotimizada. Carga parasitária, dano tecidual e resposta immune variaram mesmo em cães com quadros clínicos similares, não sendo, portanto, a análise clínica um bom parâmetro para avaliação de susceptibilidade animal à LVVisceral leishmaniasis (VL) is a zoonotic disease with a canine urban reservoir in South America. Dogs from an endemic area within Brazil, which were naturally infected with Leishmania infantum, and those presenting severe clinical (SC), mild, or no clinical (MNC) disease, were evaluated. Parasite load, histopathology, and cytokine and iNOS mRNA expressions were assessed in the spleen and liver in order to determine the potential markers for disease susceptibility or resistance. As a result, dogs with both SC and MNC had high parasite loads; IFN-γ was the most expressive cytokine in both organs, along with IL-6 and IL-4 being detected in the spleen and liver, and IL-10 only in liver. The hepatic tissue presented higher medians for IFN-γ and IL-10, and was the main organ to produce cytokines with hepatic IL-10 suggesting a regulatory follow up. Granulomas were detected in both organs; however, when absent in spleen, they were associated with elevated IL-6 levels, thus highlighting the anti-inflammatory role of IL-6. Microscopic lesions in the spleen were predominantly characterized by an extensively disorganized white pulp and splenic response was suggested as sub optimized. Parasite load, tissue damage, and immunological response may vary in the dogs with similar clinical symptoms, which may not be a good parameter for assessing the animal’s susceptibility to VL

Cytokine and iNOS profiles in lymph nodes of dogs naturally infected with Leishmania infantum and their association with the parasitic DNA load and clinical and histopathological features

Submitted by Janaína Nascimento ([email protected]) on 2019-01-21T11:42:52Z No. of bitstreams: 1 ve_Vasconcelos_Tassia_etal_INI_2016.pdf: 1709103 bytes, checksum: 905265d15c4a86726a30a76d3b6a649f (MD5)Approved for entry into archive by Janaína Nascimento ([email protected]) on 2019-01-22T10:27:57Z (GMT) No. of bitstreams: 1 ve_Vasconcelos_Tassia_etal_INI_2016.pdf: 1709103 bytes, checksum: 905265d15c4a86726a30a76d3b6a649f (MD5)Made available in DSpace on 2019-01-22T10:27:57Z (GMT). No. of bitstreams: 1 ve_Vasconcelos_Tassia_etal_INI_2016.pdf: 1709103 bytes, checksum: 905265d15c4a86726a30a76d3b6a649f (MD5) Previous issue date: 2016Universidade Federal Fluminense. Pós-graduação em Medicina Veterinária. Niterói, RJ, Brasil.Institut Pasteur. Unit Cytokines and Inflammation. Department of Infection & Epidemiology. Paris, France.Institut Pasteur. Unit Cytokines and Inflammation. Department of Infection & Epidemiology. Paris, France.Universidade Federal Fluminense. Pós-graduação em Medicina Veterinária. Niterói, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Vigilância em Leishmanioses. Rio de Janeiro, RJ, Brasil.Universidade Federal de São João del-Rei. Campus Centro-Oeste “Dona Lindu”. Divinópolis, MG, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ, Brasil.In South America, visceral leishmaniasis is a zoonotic disease with severe evolution characteristics in humans, and dogs are its main reservoir. In this context, this study aimed to evaluate the clinical status of dogs from a Brazilian endemic area naturally, at Barra Mansa municipality, infected with Leishmania infantum, in conjunction with their histopathological profile and, in order to determine possible markers of susceptibility or resistance to the disease, parasitic DNA load, cytokine and iNOS mRNA expression profiles were investigated in lymph nodes. High levels of IFN-ɣ and IL-6 mRNA were detected. Both IFN-ɣ and IL-6 mRNA were associated with disorganization of the corticomedullary region. IFN-ɣ and TNF-α mRNA were associated with the absence of follicular hyperplasia. The regulatory pathway was remarkable with IL-10 mRNA detection and its significant association with the severity of the disease. Plasmacytosis and sinus histiocytosis were associated with high loads of parasitic DNA, but there was no significant association between the parasite DNA load and animal clinical alterations. Since high parasitic loads were found in animals with or without symptoms, clinical examination cannot be considered as a criterion for disease susceptibility assessment

Gene Co-expression Analysis Indicates Potential Pathways and Regulators of Beef Tenderness in Nellore Cattle

Beef tenderness, a complex trait affected by many factors, is economically important to beef quality, industry, and consumer’s palatability. In this study, RNA-Seq was used in network analysis to better understand the biological processes that lead to differences in beef tenderness. Skeletal muscle transcriptional profiles from 24 Nellore steers, selected by extreme estimated breeding values (EBVs) for shear force after 14 days of aging, were analyzed and 22 differentially expressed transcripts were identified. Among these were genes encoding ribosomal proteins, glutathione transporter ATP-binding cassette, sub-family C (CFTR/MRP), member 4 (ABCC4), and synaptotagmin IV (SYT4). Complementary co-expression analyses using Partial Correlation with Information Theory (PCIT), Phenotypic Impact Factor (PIF) and the Regulatory Impact Factor (RIF) methods identified candidate regulators and related pathways. The PCIT analysis identified ubiquitin specific peptidase 2 (USP2), growth factor receptor-bound protein 10 (GBR10), anoctamin 1 (ANO1), and transmembrane BAX inhibitor motif containing 4 (TMBIM4) as the most differentially hubbed (DH) transcripts. The transcripts that had a significant correlation with USP2, GBR10, ANO1, and TMBIM4 enriched for proteasome KEGG pathway. RIF analysis identified microRNAs as candidate regulators of variation in tenderness, including bta-mir-133a-2and bta-mir-22. Both microRNAs have target genes present in the calcium signaling pathway and apoptosis. PIF analysis identified myoglobin (MB), enolase 3 (ENO3), and carbonic anhydrase 3 (CA3) as potentially having fundamental roles in tenderness. Pathways identified in our study impacted in beef tenderness included: calcium signaling, apoptosis, and proteolysis. These findings underscore some of the complex molecular mechanisms that control beef tenderness in Nellore cattle.This article is published as Gonçalves TM, de Almeida Regitano LC, Koltes JE, Cesar ASM, da Silva Andrade SC, Mourão GB, Gasparin G, Moreira GCM, Fritz-Waters E, Reecy JM and Coutinho LL (2018) Gene Co-expression Analysis Indicates Potential Pathways and Regulators of Beef Tenderness in Nellore Cattle. Front. Genet. 9:441. doi: 10.3389/fgene.2018.00441.</p