Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cães
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
dARK ID: | ark:/26339/0013000005zpp |
Texto Completo: | http://repositorio.ufsm.br/handle/1/14103 |
Resumo: | Mast cell tumors (MCTs) is the most prevalent cutaneous mesenchymal tumor of the dog. In the region of Santa Maria, RS, this neoplasm corresponds to 20% of the malignant skin tumors diagnosed in the laboratory routine. The clinical-pathological diagnosis can be made through cytology, histopathological examination and immunohistochemistry. The immunohistochemical technique is used to differentiate biologically more aggressive tumors from less aggressive tumors. However, some of the currently available immunomarkers are influenced by formalin fixation, interfering with the results. The main objective of this study was to evaluate the interference of formaldehyde in the immunostaining of KIT protein and Ki67 protein in four pre-established fixation times (24, 48, 72 and 96h) in MCTs. Two amplification systems were also used to determine possible differences in detection for each one of the antibodies studied. This study was divided into two parts: a retrospective study, analyzing tissue samples from 25 cases of biopsies without knowledge of fixation time; and a prospective study, analyzing tissue samples from 12 cases with known fixation times. In both studies, two histological grading systems (Patnaik and Kiupel), the special staining of toluidine blue and the immunohistochemistry for KIT and Ki67 proteins were applied. Additionally, in the prospective study, two amplification systems (biotinylated and not biotinylated) for the Ki67 protein and the AgNOR technique (counting of the argirophilic nucleolar organizing regions) were tested. It was observed that the fixation for up to 96h did not interfere in the immunohistochemical evaluation for the KIT protein. However, for Ki67 protein, there was marked interference when the fixation was greater than 24h. Both amplification systems showed satisfactory results. However, there were no differences and/or improvement in the results of immunostaing for Ki67 when compared the amplification systems. The additional useof the AgNOR method to evaluate the rate of cell proliferation is suggested when the fixation time of the neoplasm is unknown or greater than 24 hours. |
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Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cãesHistological, histochemical and immunohistochemistry evaluation of skin mast cell tumors in dogsCãoPeleNeoplasmaMastocitomaComportamento biológicoAgNORImuno-histoquímicaKi-67KITDogSkinNeoplasmMast cell tumorsBiological behaviorAgNORImmunohistochemistryCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAMast cell tumors (MCTs) is the most prevalent cutaneous mesenchymal tumor of the dog. In the region of Santa Maria, RS, this neoplasm corresponds to 20% of the malignant skin tumors diagnosed in the laboratory routine. The clinical-pathological diagnosis can be made through cytology, histopathological examination and immunohistochemistry. The immunohistochemical technique is used to differentiate biologically more aggressive tumors from less aggressive tumors. However, some of the currently available immunomarkers are influenced by formalin fixation, interfering with the results. The main objective of this study was to evaluate the interference of formaldehyde in the immunostaining of KIT protein and Ki67 protein in four pre-established fixation times (24, 48, 72 and 96h) in MCTs. Two amplification systems were also used to determine possible differences in detection for each one of the antibodies studied. This study was divided into two parts: a retrospective study, analyzing tissue samples from 25 cases of biopsies without knowledge of fixation time; and a prospective study, analyzing tissue samples from 12 cases with known fixation times. In both studies, two histological grading systems (Patnaik and Kiupel), the special staining of toluidine blue and the immunohistochemistry for KIT and Ki67 proteins were applied. Additionally, in the prospective study, two amplification systems (biotinylated and not biotinylated) for the Ki67 protein and the AgNOR technique (counting of the argirophilic nucleolar organizing regions) were tested. It was observed that the fixation for up to 96h did not interfere in the immunohistochemical evaluation for the KIT protein. However, for Ki67 protein, there was marked interference when the fixation was greater than 24h. Both amplification systems showed satisfactory results. However, there were no differences and/or improvement in the results of immunostaing for Ki67 when compared the amplification systems. The additional useof the AgNOR method to evaluate the rate of cell proliferation is suggested when the fixation time of the neoplasm is unknown or greater than 24 hours.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqO mastocitoma é o tumor mesenquimal cutâneo mais prevalente do cão. Na região de Santa Maria, RS, este neoplasma corresponde a 20% dos tumores malignos de pele diagnosticados na rotina laboratorial. O diagnóstico clínico-patológico pode ser realizado através da citologia, exame histopatológico e imuno-histoquímico. A técnica de imunohistoquímica é empregada para diferenciar tumores biologicamente mais agressivos de tumores menos agressivos. Entretanto, alguns dos imunomarcadores atualmente disponíveis sofrem influência pela fixação em formol, interferindo nos resultados. O principal objetivo deste estudo foi avaliar em mastocitomas cutâneos caninos a interferência do formol na imunomarcação para a proteína KIT e a proteína Ki67 em quatro tempos de fixação pré-estabelecidos (24, 48, 72 e 96h). Foram utilizados ainda, dois sistemas de amplificação para determinar possíveis diferenças na detecção para cada um dos dois anticorpos estudados. Este estudo foi dividido em duas partes: um estudo retrospectivo, analisando amostras de tecidos de 25 casos de biópsias sem conhecimento do tempo de fixação; e um estudo prospectivo, analisando amostras de tecidos de 12 casos com tempos de fixação conhecidos. Em ambos os estudos foram aplicados dois sistemas de graduação histológica (Patnaik e Kiupel), a coloração especial de Azul de Toluidina e a imuno-histoquímica para as proteínas KIT e Ki67. Adicionalmente, no estudo prospectivo, foram testados dois sistemas de amplificação para a proteína Ki67 (biotinilado e não biotinilado) e a técnica de AgNOR (contagem das regiões organizadoras nucleolares argirofílicas). Foi observado que a fixação por até 96h não interferiu na avaliação imuno-histoquímica para a proteína KIT. Porém, para a proteína Ki67, houve marcada interferência quando a fixação foi superior a 24h. Ambos os sistemas de amplificação utilizados mostraram resultados satisfatórios. Porém, não houve diferenças e/ou melhora nos resultados da imunomarcação para o Ki67, quando comparados os dois sistemas de amplificação. A utilização adicional do método histoquímico de AgNOR, para avaliar a taxa de proliferação celular, é sugerida quando o tempo de fixação do neoplasma for desconhecido ou superior a 24h.Universidade Federal de Santa MariaBrasilMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaCentro de Ciências RuraisKommers, Glaucia Denisehttp://lattes.cnpq.br/5818649889964582Fernandes, Cristina Gevehrhttp://lattes.cnpq.br/4498627593892325Flores, Mariana Martinshttp://lattes.cnpq.br/5683966482735545Santos, Alex dos2018-08-27T13:59:56Z2018-08-27T13:59:56Z2018-02-05info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/14103ark:/26339/0013000005zppporAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2022-04-06T17:16:56Zoai:repositorio.ufsm.br:1/14103Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2022-04-06T17:16:56Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cães Histological, histochemical and immunohistochemistry evaluation of skin mast cell tumors in dogs |
title |
Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cães |
spellingShingle |
Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cães Santos, Alex dos Cão Pele Neoplasma Mastocitoma Comportamento biológico AgNOR Imuno-histoquímica Ki-67 KIT Dog Skin Neoplasm Mast cell tumors Biological behavior AgNOR Immunohistochemistry CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cães |
title_full |
Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cães |
title_fullStr |
Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cães |
title_full_unstemmed |
Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cães |
title_sort |
Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cães |
author |
Santos, Alex dos |
author_facet |
Santos, Alex dos |
author_role |
author |
dc.contributor.none.fl_str_mv |
Kommers, Glaucia Denise http://lattes.cnpq.br/5818649889964582 Fernandes, Cristina Gevehr http://lattes.cnpq.br/4498627593892325 Flores, Mariana Martins http://lattes.cnpq.br/5683966482735545 |
dc.contributor.author.fl_str_mv |
Santos, Alex dos |
dc.subject.por.fl_str_mv |
Cão Pele Neoplasma Mastocitoma Comportamento biológico AgNOR Imuno-histoquímica Ki-67 KIT Dog Skin Neoplasm Mast cell tumors Biological behavior AgNOR Immunohistochemistry CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
topic |
Cão Pele Neoplasma Mastocitoma Comportamento biológico AgNOR Imuno-histoquímica Ki-67 KIT Dog Skin Neoplasm Mast cell tumors Biological behavior AgNOR Immunohistochemistry CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
Mast cell tumors (MCTs) is the most prevalent cutaneous mesenchymal tumor of the dog. In the region of Santa Maria, RS, this neoplasm corresponds to 20% of the malignant skin tumors diagnosed in the laboratory routine. The clinical-pathological diagnosis can be made through cytology, histopathological examination and immunohistochemistry. The immunohistochemical technique is used to differentiate biologically more aggressive tumors from less aggressive tumors. However, some of the currently available immunomarkers are influenced by formalin fixation, interfering with the results. The main objective of this study was to evaluate the interference of formaldehyde in the immunostaining of KIT protein and Ki67 protein in four pre-established fixation times (24, 48, 72 and 96h) in MCTs. Two amplification systems were also used to determine possible differences in detection for each one of the antibodies studied. This study was divided into two parts: a retrospective study, analyzing tissue samples from 25 cases of biopsies without knowledge of fixation time; and a prospective study, analyzing tissue samples from 12 cases with known fixation times. In both studies, two histological grading systems (Patnaik and Kiupel), the special staining of toluidine blue and the immunohistochemistry for KIT and Ki67 proteins were applied. Additionally, in the prospective study, two amplification systems (biotinylated and not biotinylated) for the Ki67 protein and the AgNOR technique (counting of the argirophilic nucleolar organizing regions) were tested. It was observed that the fixation for up to 96h did not interfere in the immunohistochemical evaluation for the KIT protein. However, for Ki67 protein, there was marked interference when the fixation was greater than 24h. Both amplification systems showed satisfactory results. However, there were no differences and/or improvement in the results of immunostaing for Ki67 when compared the amplification systems. The additional useof the AgNOR method to evaluate the rate of cell proliferation is suggested when the fixation time of the neoplasm is unknown or greater than 24 hours. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-08-27T13:59:56Z 2018-08-27T13:59:56Z 2018-02-05 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/14103 |
dc.identifier.dark.fl_str_mv |
ark:/26339/0013000005zpp |
url |
http://repositorio.ufsm.br/handle/1/14103 |
identifier_str_mv |
ark:/26339/0013000005zpp |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária Centro de Ciências Rurais |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária Centro de Ciências Rurais |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
_version_ |
1815172290310045696 |