Avaliação histológica,histoquímica e imuno-histoquímica de mastocitomas cutâneos em cães

Detalhes bibliográficos
Autor(a) principal: Santos, Alex dos
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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spelling 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
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