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<p>This study was carried out to determine the phenotype of special dog mammary gland tumours that were grown in nude mice. 26 tumours were examined by the immunohistochemical ABC-Elite protocol. The tumour tissues were labelled with following anti-human antibodies:</p><p>- AE1/AE3 (pankeratin antibody) labelled epithelial and myoepithelial cells</p><p>- CD 31 labelled endothelial cells</p><p>- desmin labelled cross-striated and smooth muscle cells</p><p>- myosin labelled cross striated muscle cells</p><p>- neurofilament (NF) labelled nerve cells</p><p>- osteopontin labelled preosteoblasts, osteoblasts and osteocytes</p><p>- p63 labelled nuclei of the myoepithelial cells</p><p>- smooth muscle actin (SMA) labelled the cytoplasm of myoepithelial cells</p><p>- type I collagen labelled the extracellular matrix in connective tissue and bone</p><p>- type II collagen labelled the extracellular matrix in cartilage</p><p>- vimentin labelled fibroblasts, fibrocytes, lipocytes, smooth muscle cells, endothelial cells, nerve cells, macrophages and myoepithelial cells</p><p>The tumours were also submitted to a double immunolabelling study using p63 and SMA.</p><p>The study could not give a final conclusion about the origin the tumours. There was still need for more research to answer that question. However, the immunohistochemical technique was analysed in detail, in order to obtain perfect labelings.</p><p>Initially, all the antibodies were tested on normal dog tissue, to acquire the best working dilutions with the lowest background problems. In the tumours, good results were obtained with these dilutions for the antibodies p63, SMA, vimentin, desmin, NF, AE1/AE3 and CD 31. Except for type I collagen, type II collagen and osteopontin that gave too much unspecific labelling of the mouse connective tissue. Even, when using the Vector® M.O.M. blocking kit, the results were still very difficult to interpretate.</p><p>The antigen retrieval methods were evaluated for all the antibodies. The antibodies p63, SMA, vimentin, desmin, AE1/AE3, myosin, neurofilament and CD 31 needed the antigen retrieval treatment. The antibodies type I collagen and type II collagen needed the treatment with the enzyme pepsin, while osteopontin did not need any pretreatment at all.</p><p>The double immunolabelling with p63 and SMA gave excellent results. Different combinations were tried out with different substrates, namely Vector® Nova RED, Vector® DAB and Vector® SG. Vector® methyl green was used as counterstaining, but it interfered with the other substrates, and better results were obtained without this counterstaining.</p>

This study was carried out to determine the phenotype of special dog mammary gland tumours that were grown in nude mice. 26 tumours were examined by the immunohistochemical ABC-Elite protocol. The tumour tissues were labelled with following anti-human antibodies: - AE1/AE3 (pankeratin antibody) labelled epithelial and myoepithelial cells - CD 31 labelled endothelial cells - desmin labelled cross-striated and smooth muscle cells - myosin labelled cross striated muscle cells - neurofilament (NF) labelled nerve cells - osteopontin labelled preosteoblasts, osteoblasts and osteocytes - p63 labelled nuclei of the myoepithelial cells - smooth muscle actin (SMA) labelled the cytoplasm of myoepithelial cells - type I collagen labelled the extracellular matrix in connective tissue and bone - type II collagen labelled the extracellular matrix in cartilage - vimentin labelled fibroblasts, fibrocytes, lipocytes, smooth muscle cells, endothelial cells, nerve cells, macrophages and myoepithelial cells The tumours were also submitted to a double immunolabelling study using p63 and SMA. The study could not give a final conclusion about the origin the tumours. There was still need for more research to answer that question. However, the immunohistochemical technique was analysed in detail, in order to obtain perfect labelings. Initially, all the antibodies were tested on normal dog tissue, to acquire the best working dilutions with the lowest background problems. In the tumours, good results were obtained with these dilutions for the antibodies p63, SMA, vimentin, desmin, NF, AE1/AE3 and CD 31. Except for type I collagen, type II collagen and osteopontin that gave too much unspecific labelling of the mouse connective tissue. Even, when using the Vector® M.O.M. blocking kit, the results were still very difficult to interpretate. The antigen retrieval methods were evaluated for all the antibodies. The antibodies p63, SMA, vimentin, desmin, AE1/AE3, myosin, neurofilament and CD 31 needed the antigen retrieval treatment. The antibodies type I collagen and type II collagen needed the treatment with the enzyme pepsin, while osteopontin did not need any pretreatment at all. The double immunolabelling with p63 and SMA gave excellent results. Different combinations were tried out with different substrates, namely Vector® Nova RED, Vector® DAB and Vector® SG. Vector® methyl green was used as counterstaining, but it interfered with the other substrates, and better results were obtained without this counterstaining.

Made available in DSpace on 2014-06-11T19:27:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-01Bitstream added on 2014-06-13T20:57:22Z : No. of bitstreams: 1 magalhaes_gm_me_jabo.pdf: 1621150 bytes, checksum: b5a01c9394af10e93a8a4a2595f1190d (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / As neoplasias mamárias caninas são as mais frequentes nessa espécie e também são comuns em mulheres. Um dos principais fatores prognósticos dessa enfermidade é a presença ou ausência de metástase em linfonodos. A metástase é um mecanismo complexo que envolve vários fatores. Em mulheres sabe-se que a molécula de adesão CD44 está relacionada com invasão e metástase. Este trabalho teve como objetivos: avaliar a imunomarcação de CD44 nas neoplasias mamárias malignas da cadela, com e sem metástase em linfonodos regionais; associá-la como fator prognóstico; na detecção precoce de metástase; relacioná-la com a imunomarcação de MMP-9, E-caderina e VEGF e associar com a sobrevida das cadelas. Compuseram os grupos experimentais, cadelas com tumor mamário, com (Grupo M) ou sem metástase (Grupo N) detectável em linfonodos e um grupo controle composto por tecido mamário normal. Não houve predileção por localização mamária, mas utilizou-se mais as mamas inguinais e linfonodos inguinais. As neoplasias mamárias foram classificadas de acordo com a Organização Mundial de Saúde. Para isso utilizou-se a técnica de imuno-histoquímica, em amostras incluídas em parafina. Para a determinação da porcentagem de imunomarcação considerou-se somente as células epiteliais neoplásicas. Para o anticorpo CD44 contou-se as marcações em linfócitos T nos linfonodos dos dois grupos Observou-se aumento significativo na marcação do CD44 do sítio primário do tumor para a metástase, assim como nas marcações de MMP-9 e E-caderina. As marcações em linfócitos T foram maiores no grupo N e menores no grupo M. O tipo histopatológico mais comum foi o carcinoma simples túbulo papilífero. A raça predominante no grupo das metástases foi Teckel, com idade média de 9,4 anos, e com tempo de sobrevida de sete meses. Concluiu-se... / Mammary neoplasia are the most frequently type of cancer in bitches and also in women. One of the main prognostic factors of this desease is the presence or absent of lymphonodes metastasis. Metastasis is a complex mechanism which involve multiple factors. At women, researchs have shown that the CD44, a adhesion molecule, is related with invasion and metastasis. This research looked for as objectives: Evaluation of CD44 in malignant mammary neoplasia at bitches with and without lymphonodes metastasis; link CD44 and prognosis; CD44 and early detection of metastasis; link CD44 with MMP-9; E-cadherin and VEGF; and associate with the survival of bitches. Experimental groups were arranged with bitches suffering from mammary neoplasia and divided into three groups: Group one(M) were compounded by bitches with visible metastasis in the lymphonodes; Group two(N) were compounded by bitches without visible metastasis in the lymphonodes;Group three or control compounded by bitches with normal mammary tissue. During the research did not have predilection about the mammary localization, despite inguinal breast and lymphonodes were more collected. Mammary neoplasias were classified according to the OMS norms. For this we used the technique of immunohistochemistry in paraffin sections and counted only epithelial cancer cells by method of percentage of labeled cells. Only antibody to CD44 told that the markings on T lymphocytes in lymph nodes of the two groups observed a significant increase in CD44 marking the site of the primary tumor to metastasis, as well as the markings of MMP-9 and E-cadherin. The markings on T lymphocytes were higher in group N and lower in group M. The most common pathological type was the simple tubular papillary carcinoma. The xiii predominant race in the group of metastasis was Daschshund, mean age 9,4 years, and survival... (Complete abstract click electronic access below)

Orientador: Antonio Carlos Alessi / Banca: Felipe Augusto Ruiz Sueiro / Banca: Rosemeri de Oliveira Vasconcelos / Resumo: As neoplasias mamárias caninas são as mais frequentes nessa espécie e também são comuns em mulheres. Um dos principais fatores prognósticos dessa enfermidade é a presença ou ausência de metástase em linfonodos. A metástase é um mecanismo complexo que envolve vários fatores. Em mulheres sabe-se que a molécula de adesão CD44 está relacionada com invasão e metástase. Este trabalho teve como objetivos: avaliar a imunomarcação de CD44 nas neoplasias mamárias malignas da cadela, com e sem metástase em linfonodos regionais; associá-la como fator prognóstico; na detecção precoce de metástase; relacioná-la com a imunomarcação de MMP-9, E-caderina e VEGF e associar com a sobrevida das cadelas. Compuseram os grupos experimentais, cadelas com tumor mamário, com (Grupo M) ou sem metástase (Grupo N) detectável em linfonodos e um grupo controle composto por tecido mamário normal. Não houve predileção por localização mamária, mas utilizou-se mais as mamas inguinais e linfonodos inguinais. As neoplasias mamárias foram classificadas de acordo com a Organização Mundial de Saúde. Para isso utilizou-se a técnica de imuno-histoquímica, em amostras incluídas em parafina. Para a determinação da porcentagem de imunomarcação considerou-se somente as células epiteliais neoplásicas. Para o anticorpo CD44 contou-se as marcações em linfócitos T nos linfonodos dos dois grupos Observou-se aumento significativo na marcação do CD44 do sítio primário do tumor para a metástase, assim como nas marcações de MMP-9 e E-caderina. As marcações em linfócitos T foram maiores no grupo N e menores no grupo M. O tipo histopatológico mais comum foi o carcinoma simples túbulo papilífero. A raça predominante no grupo das metástases foi Teckel, com idade média de 9,4 anos, e com tempo de sobrevida de sete meses. Concluiu-se... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Mammary neoplasia are the most frequently type of cancer in bitches and also in women. One of the main prognostic factors of this desease is the presence or absent of lymphonodes metastasis. Metastasis is a complex mechanism which involve multiple factors. At women, researchs have shown that the CD44, a adhesion molecule, is related with invasion and metastasis. This research looked for as objectives: Evaluation of CD44 in malignant mammary neoplasia at bitches with and without lymphonodes metastasis; link CD44 and prognosis; CD44 and early detection of metastasis; link CD44 with MMP-9; E-cadherin and VEGF; and associate with the survival of bitches. Experimental groups were arranged with bitches suffering from mammary neoplasia and divided into three groups: Group one(M) were compounded by bitches with visible metastasis in the lymphonodes; Group two(N) were compounded by bitches without visible metastasis in the lymphonodes;Group three or control compounded by bitches with normal mammary tissue. During the research did not have predilection about the mammary localization, despite inguinal breast and lymphonodes were more collected. Mammary neoplasias were classified according to the OMS norms. For this we used the technique of immunohistochemistry in paraffin sections and counted only epithelial cancer cells by method of percentage of labeled cells. Only antibody to CD44 told that the markings on T lymphocytes in lymph nodes of the two groups observed a significant increase in CD44 marking the site of the primary tumor to metastasis, as well as the markings of MMP-9 and E-cadherin. The markings on T lymphocytes were higher in group N and lower in group M. The most common pathological type was the simple tubular papillary carcinoma. The xiii predominant race in the group of metastasis was Daschshund, mean age 9,4 years, and survival... (Complete abstract click electronic access below) / Mestre