Adenosquamous carcinoma in the palate of a dog: case report

Adenosquamous carcinoma is characterized by the simultaneous presence of squamous and glandular components. This article reports a case of adenosquamous carcinoma in the oral mucosa in a dog. Microscopically, the tumour revealed an infiltrative epithelial proliferation with a tubular pattern and areas of squamous differentiation. In the lumina of the tubules, there were eosinophilic material (mucin). The alcian blue staining showed positive intraluminal material and the cytoplasm of neoplastic cells were immunoreactive for cytokeratin, supporting the diagnosis of adenosquamous carcinoma. The cells exhibited reduced or negative immunoreactivity for E-cadherin, which could be associated with more aggressive tumour behaviour and worse prognosis.(AU)

Sucralfate enemas reduce the oxidative tissue damage and preserves the contents of E-cadherin and Β-catenin in colonic mucosa without fecal stream

ABSTRACT Purpose: To evaluate the effects of sucralfate enemas in tissue contents of E-cadherin and ?-catenin in an experimental diversion colitis. Methods: Thirty-six male Wistar rats were submitted to a proximal colostomy and a distal mucous fistula. They were allocated into three groups: first group received daily saline enemas (2 mL/day) and the two other groups daily enemas with sucralfate at dosage of 1 or 2 g/kg/day, respectively. Six animals of each group were euthanized after two weeks and six animals after four weeks. The inflammation of the excluded mucosa was evaluated by histological analysis. The oxidative damage was quantified by measurement of malondialdehyde tissue levels. The expression of E-cadherin and ?-catenin was identified by immunohistochemistry, and its contents were quantified by computer-assisted image analysis. Results: Sucralfate enemas reduced inflammation in animals subjected to treatment with 2 g/kg/day by four weeks, and the levels of oxidative damage in mucosa without fecal stream irrespective of concentration and time of intervention. E-cadherin and ?-catenin content increased in segments without fecal stream in those animals subjected to treatment with sucralfate. Conclusions: Sucralfate reduces the inflammation and oxidative stress and increases the tissue content of E-cadherin and ?-catenin in colonic mucosa devoid to the fecal stream.

Prognostic impact of alterations in E-cadherin and P-cadherin expression in canine mammary tumors

A expressão de caderinas tem sido correlacionada ao desenvolvimento e agressividade de neoplasias epiteliais, entretanto, em tumores mamários caninos, seu prognóstico de importância é incerto. Devido a esse fato, a expressão das moléculas de adesão intracelular E e P-caderina e a correlação com a sobrevida global foram analisadas em 25 tumores de glândula mamária canina. A redução da expressão da caderina-E foi correlacionada com o tipo histológico, alto grau histológico e taxa de sobrevida global. A coloração de P-caderina foi maior em tumores malignos, sem relação com outras características clínico-patológicas de agressividade. Os resultados deste estudo sugerem uma relação entre menor expressão de E e P-caderina e pior prognóstico em tumores mamários caninos, incluindo menor sobrevida global.

Prognostic impact of alterations in E-cadherin and P-cadherin expression in canine mammary tumors

A expressão de caderinas tem sido correlacionada ao desenvolvimento e agressividade de neoplasias epiteliais, entretanto, em tumores mamários caninos, seu prognóstico de importância é incerto. Devido a esse fato, a expressão das moléculas de adesão intracelular E e P-caderina e a correlação com a sobrevida global foram analisadas em 25 tumores de glândula mamária canina. A redução da expressão da caderina-E foi correlacionada com o tipo histológico, alto grau histológico e taxa de sobrevida global. A coloração de P-caderina foi maior em tumores malignos, sem relação com outras características clínico-patológicas de agressividade. Os resultados deste estudo sugerem uma relação entre menor expressão de E e P-caderina e pior prognóstico em tumores mamários caninos, incluindo menor sobrevida global.(AU)

Progressive histiocytosis of non-epitheliotropic dendritic cells in a feline

Background: Histiocytic tumors in felines are nodules that commonly develop on limbs and head extremities. They can be divided into many subtypes including cutaneous histiocytoma, histiocytic sarcoma, reactive fibrohistiocytic nodule, Langerhans cell histiocytosis, and progressive feline dendritic cell. Despite the same origin, they have behaviors that differ from each other, thus it is important to confirm diagnosis with histopathological and immunohistochemical tests, because early identification can facilitate prognosis and treatment. In this study, we describe the pathological and immunohistochemical characteristics, enabling differentiation feline neoplasms derived from histiocytes. Case: A 5-year-old, crossbreed, male, feline presented with a nodulation at the base of the left ear. The mass was slow growing, partially alopecic, with no other changes associated with tumor development. The nodule was round and circumscribed, movable, with an elevated surface. He was referred for surgery and an elliptical sample around the tumor was carefully dissected. Routine histopathological evaluation was performed with hematoxylin and eosin (HE), as well as immunohistochemistry. Histopathology showed circumscribed proliferation of histiocytic cells, with abundant and eosinophilic cytoplasm. The proliferative cells were large and rounded, extending from the superficial dermis and basement membrane to the deep dermis. At the extremities, some cells had visible vacuoles. Mitotic activity ranged from 3 to 4 mitoses per field in 40x magnification. Immunohistochemistry showed positive staining for histocompatibility complex MCII and lysozyme antibodies, marking histiocytic cells. Labeling was positive for CD20 in cells of lymphoid lineage B and negative for E-cadherin. Histiocytic cells did not invade the epidermis; hence, proliferation was classified as nonepitheliotropic. These methods contribute to the literature regarding the...

Progressive histiocytosis of non-epitheliotropic dendritic cells in a feline

Background: Histiocytic tumors in felines are nodules that commonly develop on limbs and head extremities. They can be divided into many subtypes including cutaneous histiocytoma, histiocytic sarcoma, reactive fibrohistiocytic nodule, Langerhans cell histiocytosis, and progressive feline dendritic cell. Despite the same origin, they have behaviors that differ from each other, thus it is important to confirm diagnosis with histopathological and immunohistochemical tests, because early identification can facilitate prognosis and treatment. In this study, we describe the pathological and immunohistochemical characteristics, enabling differentiation feline neoplasms derived from histiocytes. Case: A 5-year-old, crossbreed, male, feline presented with a nodulation at the base of the left ear. The mass was slow growing, partially alopecic, with no other changes associated with tumor development. The nodule was round and circumscribed, movable, with an elevated surface. He was referred for surgery and an elliptical sample around the tumor was carefully dissected. Routine histopathological evaluation was performed with hematoxylin and eosin (HE), as well as immunohistochemistry. Histopathology showed circumscribed proliferation of histiocytic cells, with abundant and eosinophilic cytoplasm. The proliferative cells were large and rounded, extending from the superficial dermis and basement membrane to the deep dermis. At the extremities, some cells had visible vacuoles. Mitotic activity ranged from 3 to 4 mitoses per field in 40x magnification. Immunohistochemistry showed positive staining for histocompatibility complex MCII and lysozyme antibodies, marking histiocytic cells. Labeling was positive for CD20 in cells of lymphoid lineage B and negative for E-cadherin. Histiocytic cells did not invade the epidermis; hence, proliferation was classified as nonepitheliotropic. These methods contribute to the literature regarding the...(AU)

Avaliação clínica, histopatológica e imunohistoquímica da histiocitose cutânea reativa em canino na Amazônia Ocidental / Clinical, histopathological and immunohistochemistry evaluation of reactive cutaneous histiocytosis in canine in western Amazonia

Background: Canine reactive cutaneous histiocytosis (RCH) is an immuneproliferative disease of skin histiocytes and is uncommon in occurrence. Its description in the literature is scarce and clinical studies are limited by the insufficient characterization of the patients’ pathological findings. The objective of this report is to describe the clinical, histological and immunohistochemical findings of a case of canine reactive cutaneous histiocytosis in the state of Acre, Amazonia, Brazil. Case: It was attended at the Federal University of Acre, a 7-year-old male American pit bull terrier dog with nodular, allopecic and ulcerated lesions in the dorsal region of the ear, with purulent discharge and exacerbated painful tenderness. The animal was domiciled on the bank of the river Acre, municipality of Rio Branco, state of Acre, and suffered frequent parasitism by sandflies, especially in the head region. In the histopathological evaluation, hyperplastic cells were found, a large ulcerated area with the presence of fibrin and neutrophilic infiltrate in the epidermis. In the dermal layer, an inflammatory reaction pattern was identified, with the presence of fibrous connective tissue, dilated blood vessels and edema, however little defined. There was an intense presence of histiocytes with anisocytosis, in addition to neutrophils, plasma cells and lymphocytes in the perivascular and perianexal region. In immunohistochemistry, lysozyme and cell markers CD1a and Thy1 were detected, but negative result for E-cadherin and CD11d. The immunosuppressive therapy indicated with prednisolone, plus cephalexin for secondary infections and topical treatment, with clinical remission within two years. Discussion: Although the etiopathogenesis of RCH is poorly understood, it is believed that, in addition to the genetic factor, the disease is triggered by an antigenic trigger...

Avaliação clínica, histopatológica e imunohistoquímica da histiocitose cutânea reativa em canino na Amazônia Ocidental / Clinical, histopathological and immunohistochemistry evaluation of reactive cutaneous histiocytosis in canine in western Amazonia

Background: Canine reactive cutaneous histiocytosis (RCH) is an immuneproliferative disease of skin histiocytes and is uncommon in occurrence. Its description in the literature is scarce and clinical studies are limited by the insufficient characterization of the patients pathological findings. The objective of this report is to describe the clinical, histological and immunohistochemical findings of a case of canine reactive cutaneous histiocytosis in the state of Acre, Amazonia, Brazil. Case: It was attended at the Federal University of Acre, a 7-year-old male American pit bull terrier dog with nodular, allopecic and ulcerated lesions in the dorsal region of the ear, with purulent discharge and exacerbated painful tenderness. The animal was domiciled on the bank of the river Acre, municipality of Rio Branco, state of Acre, and suffered frequent parasitism by sandflies, especially in the head region. In the histopathological evaluation, hyperplastic cells were found, a large ulcerated area with the presence of fibrin and neutrophilic infiltrate in the epidermis. In the dermal layer, an inflammatory reaction pattern was identified, with the presence of fibrous connective tissue, dilated blood vessels and edema, however little defined. There was an intense presence of histiocytes with anisocytosis, in addition to neutrophils, plasma cells and lymphocytes in the perivascular and perianexal region. In immunohistochemistry, lysozyme and cell markers CD1a and Thy1 were detected, but negative result for E-cadherin and CD11d. The immunosuppressive therapy indicated with prednisolone, plus cephalexin for secondary infections and topical treatment, with clinical remission within two years. Discussion: Although the etiopathogenesis of RCH is poorly understood, it is believed that, in addition to the genetic factor, the disease is triggered by an antigenic trigger...(AU)

CD45+, CD68+ and E-cadherin+ Expressions in Skin Dogs Naturally Infected by Leishmania infantum

Background: In canine leishmaniasis (CanL), infection occurs through phlebotomine vectors that inoculate the protozoan Leishmania infantum into the skin that infected macrophages and activated dendritic cells (CD). Dogs with CanL present variable clinical manifestations, being common the presence of cutaneous lesions. The aim of this study was to evaluate the expression of CD45+, CD68+ and E-cadherin+  associating the skin sentinels cells and to compare the clinical-dermatological manifestations in the skin of dogs naturally infected by L. infantum.Materials, Methods & Results: Dogs infected (n = 22) by L. infantum were divided into asymptomatic group (AD, n = 9), and symptomatic group (SD, n = 13), according criteria based on the presence or absence of skin changes. Dogs non-infected (CD, n = 5) were included as control group. Samples of skin biopsies collected from scapular region were processed by routine histology and labeled by immunohistochemistry with monoclonal antibodies against CD45+, CD68+ and E-cadherin+, and were described as none, mild, moderate and intense. SD presented keratoconjunctivitis, onychogryphose, lichenification, depigmentation, alopecia, hypotrichosis, erythematous dermatitis, exfoliative dermatitis, ulcerative dermatitis and crusted dermatitis, and the frequency these alterations was expressed as percentage. The results of hematological and bioch

Histological evaluation and E-cadherin and ß-catenin expression in kidney of dogs submitted to renal ischemia and reperfusion after chlorpromazine administration / Avaliação histológica e imunomarcação de E-caderina e ß-catenina em rins de cães submetidos a isquemia e referfusão renal após administração de clorpromazina

Renal ischemia can be associated with some urological procedures, such as renovascular surgery or kidney transplantation, that are often followed by acute renal failure. The aim of this study was to verify the E-cadherin and ß-catenin localization in canine kidney in different times of renal ischemia and reperfusion after chlorpromazine application. Twelve dogs were randomly distributed equally into two groups. GroupA with ischemia and reperfusion without chlorpromazine and groupB with ischemia and reperfusion treated by chlorpromazine. GroupB received intravenous chlorpromazine, 15 min before the artery obstruction, which lasted 1 hour. After this period, the clamps in the renal arteries were released and the organ remained in reperfusion for 2 hours. In each group, anti-E-cadherin and anti-ß-catenin antibodies were made in six tissue samples from renal parenchyma. E-cadherin and ß-catenin are differentially expressed in segments from cortex and medulla in dog's kidneys and the use of chlorpromazine did not alter the expression of both proteins. Occlusion of the left renal artery in dogs causes morphological alterations mainly in proximal convoluted tubules, beginning 30min after the start of ischemia and being aggravated after two hours of reperfusion. These results reveal that chlorpromazine did not change kidneys' histological aspect nor E-cadherin and ß-catenin expression.(AU)

A lesão renal isquêmica pode estar associada a procedimentos urológicos, tais como cirurgia renovascular, cirurgia renal extracorpórea ou transplante renal. Essa injúria, muitas vezes, é seguida de insuficiência renal aguda. O objetivo deste trabalho foi observar a localização da E-caderina e da ß-catenina em rim de cães, além de relacionar a expressão dessas proteínas das junções de aderência em diferentes tempos de isquemia e reperfusão com ou sem a aplicação de clorpromazina. Para tanto, foram utilizados 12 cães, distribuídos aleatoriamente em dois grupos de seis indivíduos: grupo A, com isquemia e reperfusão sem tratamento por clorpromazina, e o grupo B, com isquemia e reperfusão tratado por clorpromazina. No procedimento cirúrgico, foi realizada uma incisão paracostal esquerda para identificação e isolamento do rim esquerdo e da artéria renal esquerda. Após o isolamento da artéria, os animais de todos os grupos tiveram o vaso ocluído. Os animais do grupo B receberam clorpromazina via endovenosa, na dose de 5mg/kg, 15min antes da clampagem do vaso, que durou uma hora. Após este período, as artérias renais foram desobstruídas e os órgãos permaneceram em reperfusão por duas horas. Em cada grupo, foram extraídas seis amostras de parênquima renal, com utilização de agulha tru-cut, para marcação com anticorpos anti-E-caderina e anti-ß-catenina por meio de imunoistoquímica. E-caderina e ß-catenina são diferencialmente expressas em segmentos do córtex e da medula em rim de cães e o uso da clorpromazina não alterou a expressão das duas proteínas.(AU)

Histological evaluation and E-cadherin and ß-catenin expression in kidney of dogs submitted to renal ischemia and reperfusion after chlorpromazine administration / Avaliação histológica e imunomarcação de E-caderina e ß-catenina em rins de cães submetidos a isquemia e referfusão renal após administração de clorpromazina

Renal ischemia can be associated with some urological procedures, such as renovascular surgery or kidney transplantation, that are often followed by acute renal failure. The aim of this study was to verify the E-cadherin and ß-catenin localization in canine kidney in different times of renal ischemia and reperfusion after chlorpromazine application. Twelve dogs were randomly distributed equally into two groups. GroupA with ischemia and reperfusion without chlorpromazine and groupB with ischemia and reperfusion treated by chlorpromazine. GroupB received intravenous chlorpromazine, 15 min before the artery obstruction, which lasted 1 hour. After this period, the clamps in the renal arteries were released and the organ remained in reperfusion for 2 hours. In each group, anti-E-cadherin and anti-ß-catenin antibodies were made in six tissue samples from renal parenchyma. E-cadherin and ß-catenin are differentially expressed in segments from cortex and medulla in dog's kidneys and the use of chlorpromazine did not alter the expression of both proteins. Occlusion of the left renal artery in dogs causes morphological alterations mainly in proximal convoluted tubules, beginning 30min after the start of ischemia and being aggravated after two hours of reperfusion. These results reveal that chlorpromazine did not change kidneys' histological aspect nor E-cadherin and ß-catenin expression.(AU)

A lesão renal isquêmica pode estar associada a procedimentos urológicos, tais como cirurgia renovascular, cirurgia renal extracorpórea ou transplante renal. Essa injúria, muitas vezes, é seguida de insuficiência renal aguda. O objetivo deste trabalho foi observar a localização da E-caderina e da ß-catenina em rim de cães, além de relacionar a expressão dessas proteínas das junções de aderência em diferentes tempos de isquemia e reperfusão com ou sem a aplicação de clorpromazina. Para tanto, foram utilizados 12 cães, distribuídos aleatoriamente em dois grupos de seis indivíduos: grupo A, com isquemia e reperfusão sem tratamento por clorpromazina, e o grupo B, com isquemia e reperfusão tratado por clorpromazina. No procedimento cirúrgico, foi realizada uma incisão paracostal esquerda para identificação e isolamento do rim esquerdo e da artéria renal esquerda. Após o isolamento da artéria, os animais de todos os grupos tiveram o vaso ocluído. Os animais do grupo B receberam clorpromazina via endovenosa, na dose de 5mg/kg, 15min antes da clampagem do vaso, que durou uma hora. Após este período, as artérias renais foram desobstruídas e os órgãos permaneceram em reperfusão por duas horas. Em cada grupo, foram extraídas seis amostras de parênquima renal, com utilização de agulha tru-cut, para marcação com anticorpos anti-E-caderina e anti-ß-catenina por meio de imunoistoquímica. E-caderina e ß-catenina são diferencialmente expressas em segmentos do córtex e da medula em rim de cães e o uso da clorpromazina não alterou a expressão das duas proteínas.(AU)

CD45+, CD68+ and E-cadherin+ expressions in skin dogs naturally infected by Leishmania infantum

Background: In canine leishmaniasis (CanL), infection occurs through phlebotomine vectors that inoculate the protozoan Leishmania infantum into the skin that infected macrophages and activated dendritic cells (CD). Dogs with CanL present variable clinical manifestations, being common the presence of cutaneous lesions. The aim of this study was to evaluate the expression of CD45+ , CD68+ and E-cadherin+ associating the skin sentinels cells and to compare the clinical-dermatological manifestations in the skin of dogs naturally infected by L. infantum. Materials, Methods & Results: Dogs infected (n = 22) by L. infantum were divided into asymptomatic group (AD, n = 9), and symptomatic group (SD, n = 13), according criteria based on the presence or absence of skin changes. Dogs non-infected (CD, n = 5) were included as control group. Samples of skin biopsies collected from scapular region were processed by routine histology and labeled by immunohistochemistry with monoclonal antibodies against CD45+ , CD68+ and E-cadherin+ , and were described as none, mild, moderate and intense. SD presented keratoconjunctivitis, onychogryphose, lichenification, depigmentation, alopecia, hypotrichosis, erythematous dermatitis, exfoliative dermatitis, ulcerative dermatitis and crusted dermatitis, and the frequency these alterations was expressed as percentage. The results of hematological and [...](AU)

CD45+, CD68+ and E-cadherin+ expressions in skin dogs naturally infected by Leishmania infantum

Background: In canine leishmaniasis (CanL), infection occurs through phlebotomine vectors that inoculate the protozoan Leishmania infantum into the skin that infected macrophages and activated dendritic cells (CD). Dogs with CanL present variable clinical manifestations, being common the presence of cutaneous lesions. The aim of this study was to evaluate the expression of CD45+ , CD68+ and E-cadherin+ associating the skin sentinels cells and to compare the clinical-dermatological manifestations in the skin of dogs naturally infected by L. infantum. Materials, Methods & Results: Dogs infected (n = 22) by L. infantum were divided into asymptomatic group (AD, n = 9), and symptomatic group (SD, n = 13), according criteria based on the presence or absence of skin changes. Dogs non-infected (CD, n = 5) were included as control group. Samples of skin biopsies collected from scapular region were processed by routine histology and labeled by immunohistochemistry with monoclonal antibodies against CD45+ , CD68+ and E-cadherin+ , and were described as none, mild, moderate and intense. SD presented keratoconjunctivitis, onychogryphose, lichenification, depigmentation, alopecia, hypotrichosis, erythematous dermatitis, exfoliative dermatitis, ulcerative dermatitis and crusted dermatitis, and the frequency these alterations was expressed as percentage. The results of hematological and [...]

Imunomarcação da transição epitélio-mesenquima na neoplasia mamária primária de cadelas e sua metástase em linfonodo / Epithelial to mesenchymal transition immunostaining in canine primary mammary tumor and lymph node

Os carcinomas mamários em cães apresentam alta capacidade metastática o que confere menor sobrevida para os pacientes com este tipo de neoplasia. O fenótipo transição epitélio-mesênquima, caracterizado pela troca dos filamentos intermediários de citoqueratina por vimentina, além da perda da proteína de adesão entre células (E-caderina) está relacionado com a maior ocorrência de metástase. Diante disto, objetivou-se avaliar, por meio de imunomarcações, a expressão de vimentina, citoqueratina e E-caderina nos tumores mamários caninos e suas metástases em linfonodo, a fim de avaliar o comportamento celular frente a esta neoplasia. Foram analisados cinco casos de neoplasias mamárias primárias caninas e suas respectivas metástases em linfonodos. Foram comparadas as médias de imunomarcações do grupo de neoplasias primárias com as médias do grupo metástase. Não houve diferença estatística nas imunomarcações da citoqueratina (p=0,1407) e E-caderina (p= 0,312) entre os grupos, apesar da média de expressão da E-caderina ter sido maior no grupo de metástases. A expressão da vimentina foi maior nos sítios das metástases (p=0,0462). Conclui-se que a expressão de vimentina aumenta no foco da metástase em relação aos seus respectivos tumores primários mamários caninos, caracterizando alteração estrutural celular, conferindo um fenótipo transição epitélio-mesênquima. Além da E-caderina apresentar fortes indícios de aumento no foco da metástase caracterizando maior adesão.(AU)

Mammary carcinomas in dogs have a high metastatic capacity which gives a shorter survival rate for patients with this type of tumor. The epithelial-mesenchymal transition phenotype, characterized by the trade of intermediary filaments of cytokeratin by vimentin, also by the loss of the adhesion protein between cells (E-cadherin) is associated with metastasis. Due to this fact, it was aimed to evaluate, by immunostaining, the expression of vimentin, cytokeratin and E-cadherin in canine mammary tumors and the metastasis in lymph node, in order to assess the cell behavior when facing this cancer. Five cases of canine mammary tumors and metastasis in lymph node were evaluated. The averages of immunostainings of the group of primary neoplasms were compared with the averages of the lymph node group. The results showed that immunostaining for cytokeratins (p=0,1407) and E-caderina (p=0,312) were not significant between the groups, despite the expression mean of cadherin was higher in the metastase group. The expression of vimentin (p=0,04) was greater at sites of metastases. It is concluded that the expression of vimentin increases in the focus of the metastase in relation to their respective primary canine mammary tumors, characterizing cellular structural alteration, conferring a transient epithelial-mesenchymal phenotype. And cadherin present strong evidence of increased focus on metastasis characterizing increased adhesion.(AU)

Imunomarcação da transição epitélio-mesenquima na neoplasia mamária primária de cadelas e sua metástase em linfonodo / Epithelial to mesenchymal transition immunostaining in canine primary mammary tumor and lymph node

Os carcinomas mamários em cães apresentam alta capacidade metastática o que confere menor sobrevida para os pacientes com este tipo de neoplasia. O fenótipo transição epitélio-mesênquima, caracterizado pela troca dos filamentos intermediários de citoqueratina por vimentina, além da perda da proteína de adesão entre células (E-caderina) está relacionado com a maior ocorrência de metástase. Diante disto, objetivou-se avaliar, por meio de imunomarcações, a expressão de vimentina, citoqueratina e E-caderina nos tumores mamários caninos e suas metástases em linfonodo, a fim de avaliar o comportamento celular frente a esta neoplasia. Foram analisados cinco casos de neoplasias mamárias primárias caninas e suas respectivas metástases em linfonodos. Foram comparadas as médias de imunomarcações do grupo de neoplasias primárias com as médias do grupo metástase. Não houve diferença estatística nas imunomarcações da citoqueratina (p=0,1407) e E-caderina (p= 0,312) entre os grupos, apesar da média de expressão da E-caderina ter sido maior no grupo de metástases. A expressão da vimentina foi maior nos sítios das metástases (p=0,0462). Conclui-se que a expressão de vimentina aumenta no foco da metástase em relação aos seus respectivos tumores primários mamários caninos, caracterizando alteração estrutural celular, conferindo um fenótipo transição epitélio-mesênquima. Além da E-caderina apresentar fortes indícios de aumento no foco da metástase caracterizando maior adesão.(AU)

Mammary carcinomas in dogs have a high metastatic capacity which gives a shorter survival rate for patients with this type of tumor. The epithelial-mesenchymal transition phenotype, characterized by the trade of intermediary filaments of cytokeratin by vimentin, also by the loss of the adhesion protein between cells (E-cadherin) is associated with metastasis. Due to this fact, it was aimed to evaluate, by immunostaining, the expression of vimentin, cytokeratin and E-cadherin in canine mammary tumors and the metastasis in lymph node, in order to assess the cell behavior when facing this cancer. Five cases of canine mammary tumors and metastasis in lymph node were evaluated. The averages of immunostainings of the group of primary neoplasms were compared with the averages of the lymph node group. The results showed that immunostaining for cytokeratins (p=0,1407) and E-caderina (p=0,312) were not significant between the groups, despite the expression mean of cadherin was higher in the metastase group. The expression of vimentin (p=0,04) was greater at sites of metastases. It is concluded that the expression of vimentin increases in the focus of the metastase in relation to their respective primary canine mammary tumors, characterizing cellular structural alteration, conferring a transient epithelial-mesenchymal phenotype. And cadherin present strong evidence of increased focus on metastasis characterizing increased adhesion.(AU)

CORRELAÇÃO CLÍNICO PATOLÓGICA ENTRE AS IMUNOMARCAÇÕES DE E-CADERINA E RECEPTORES DE ESTRÓGENO NOS TUMORES DE MAMA EM CADELAS

O objetivo do trabalho é estabelecer uma correlação de variáveis clínicas com as neoplasias mamárias encontradas e sua correlação sobre marcadores tumorais, podendo assim, demandar um tratamento melhor estabelecido. A relação entre o receptor de estrogênio (ER) e a expressão epitelial da caderina (E-), detectada por características clínico-patológicas e métodos imunohistoquímicos, foi avaliada em 111 tumores mamários caninos. A maioria (n = 95; 85,58%) dos tumores eram histologicamente malignos e subclassificados como grau I (n = 42), grau II (n = 31) ou grau III (n = 22). Houve uma associação estatística significativa entre malignidade grau III e imunomarcação negativa para estrogênio (p = 0,01). Nenhuma associação significativa pôde ser estabelecida entre a classificação histológica (neoplasia, benigna ou maligna) e imunomarcação para ER e caderina-E. Vinte e um (22,10 %) de 95 tumores mamários malignos metastatizaram. O carcinoma mamário simples (n = 9) foi significativamente associado à expressão ER negativa (p = 0,03), sugerindo um prognóstico ruim. Esses achados enfatizam a necessidade de melhorar os fatores prognósticos nos tumores mamários caninos

The relationship between the oestrogen receptor (ER) and epithelial (E-) cadherin expression, as detected by clinicopathological features and immunohistochemical methods, was evaluated in 111 canine mammary tumours. The majority (n = 95; 85.58%) of tumours were histologically malignant and subclassified as grade I (n = 42), grade II (n=31), or grade III (n=2022). There was a significant statistical association between grade III malignancy and negativeimmunostaining for oestrogen (p= 0.01). No significant associations could be established between histological classification (neoplasm, benign or malignant) and immunostaining for 23 ER and E-cadherin. Twenty-one (22.10%) of 95 malignant mammary tumours had metastasized. Twenty-four simple mammary carcinoma (n = 9) was significantly associated with negative ER expression (p = 0.03), suggesting a poor prognosis. These findings emphasize the necessity of improving prognostic factors in canine mammary tumours.

Progressive loss of E-cadherin immunoexpression during cervical carcinogenesis

PURPOSE: To investigate E-cadherin immunoexpression during cervical carcinogenesis. METHODS: We assessed the immunohistochemical expression of E-cadherin in squamous intraepithelial lesions (SIL - 52 cases), squamous cell carcinoma (SCC) of the uterine cervix (23 cases) and also in eight cases of cervicitis. RESULTS: The results show very different E-cadherin membrane expression levels when cervicitis (88%), SILs (73%) and SCC (17%) were compared. In SILs, higher E-cadherin loss was seen in less differentiated cells in the basal third of the epithelium. This study suggests that the absence of E-cadherin expression in the membrane is a molecular event that is observed more often in SCC of the uterine cervix than in SILs or cervicitis. CONCLUSIONS: E-cadherin is an essential molecule during the process of cervical carcinogenesis and in this context exhibits a different expression pattern according to the epithelial thickness layer. .(AU)

Papillary adenocarcinoma of nasal sinus in a dog: approach histomorphologic, immunophenotypic, and therapeutic / Adenocarcinoma papilar de seio nasal em cão: Abordagem histomorfológica, imunofenotípica e terapêutica

Background: Sinus adenocarcinomas of papillary pattern are neoplasms rarely described in dogs and most cases are characterized by slow growth, local invasion and rare cases of metastasis. Their characteristics are presence of glandular structures that may have papillary pattern, tubulo-papillary, acinar and mixed pattern. Approximately 80% of primary nasal tumors are malignant and adenocarcinomas are the most frequently histology type. This paper reports a case of papillary adenocarcinoma of nasal sinus in the dog, emphasizing the use of histochemical and immunohistochemical techniques to aiding for appropriate therapy.Case: A 14-year-old male Poodle dog, presenting seropurulent nasal discharge, frequent sneezing and an enlargement in the nasal region, was submitted to surgical excision of a tumor. Macroscopically, was observed fragment coated with soft hairy skin and shear blackish color, and some fragments associated with more firm tissues like cartilage and bone tissue. Tumor specimens were collected, fixed in 10% neutral buffered formalin solution and embedded in paraffi n. Afterwards, 4 µm histological sections were obtained and stained with HE. For immunohistochemical analysis a biotin-peroxidase system was used and secondary antibodies were identifi ed using Advance HRP. CKAE1/AE3, CK34E12, SMA, E-cadherin, Cox-2, Ki-67 and Her-2 expression were evaluated. For CK AE1AE3

Background: Sinus adenocarcinomas of papillary pattern are neoplasms rarely described in dogs and most cases are characterized by slow growth, local invasion and rare cases of metastasis. Their characteristics are presence of glandular structures that may have papillary pattern, tubulo-papillary, acinar and mixed pattern. Approximately 80% of primary nasal tumors are malignant and adenocarcinomas are the most frequently histology type. This paper reports a case of papillary adenocarcinoma of nasal sinus in the dog, emphasizing the use of histochemical and immunohistochemical techniques to aiding for appropriate therapy.Case: A 14-year-old male Poodle dog, presenting seropurulent nasal discharge, frequent sneezing and an enlargement in the nasal region, was submitted to surgical excision of a tumor. Macroscopically, was observed fragment coated with soft hairy skin and shear blackish color, and some fragments associated with more firm tissues like cartilage and bone tissue. Tumor specimens were collected, fixed in 10% neutral buffered formalin solution and embedded in paraffi n. Afterwards, 4 µm histological sections were obtained and stained with HE. For immunohistochemical analysis a biotin-peroxidase system was used and secondary antibodies were identifi ed using Advance HRP. CKAE1/AE3, CK34E12, SMA, E-cadherin, Cox-2, Ki-67 and Her-2 expression were evaluated. For CK AE1AE3

Avaliação da concordância interobservadores de critérios morfológicos no diagnóstico de tumores de glândula hepatoide em cães e determinação do valor prognóstico de aspectos clínico-patológicos e marcadores imuno-histoquímicos

Diferentes estudos têm demonstrado uma variação importante na frequência diagnóstica dos tumores de glândula hepatoide em cães, possivelmente devido à ausência de critérios morfológicos claros e bem estabelecidos em seu diagnóstico. Não estão bem definidos os critérios clínico-patológicos associados ao comportamento biológico destas neoplasias. Alguns estudos levantam a hipótese de participação do mecanismo de transição epitélio-mesenquimal devido à frequente imunoexpressão de vimentina em tumores de glândula hepatoide, porém esta hipótese permanece sem comprovação. O objetivo deste trabalho foi avaliar a concordância diagnóstica entre patologistas, assim como investigar o mecanismo de transição epitélio-mesenquimal e a taxa de proliferação celular por marcadores imuno-histoquímicos e, por fim, correlacioná-los com sobrevida para investigação de possíveis valores prognósticos. Cinco patologistas avaliaram 57 tumores de glândula hepatoide quanto ao diagnóstico e quanto a diversos critérios morfológicos e, com base nestes critérios, foram desenvolvidos algoritmos diagnósticos por meio de árvores de regressão por partição recursiva. Adicionalmente, foram investigadas as imunoexpressões de vimentina, pancitoqueratina, E-caderina e Ki-67 em 78 lesões, comparando-as entre glândulas normais, tumores benignos, tumores malignos e tumores malignos vasculoinvasivos/metastáticos. A sobrevida livre de recorrência local, recorrência da doença, sobrevida global e sobrevida doença-específica foram investigadas em 33 pacientes. Houve concordância média razoável em análise interobservador quanto ao diagnóstico (=0,54), com potencial de melhora para concordância média boa após aplicação de dois novos algoritmos diagnósticos (=0,64 e 0,65, respectivamente). Em relação aos marcadores imuno-histoquímicos, foram observadas maior expressão de vimentina e menor expressão de E-caderina em tumores malignos em comparação aos tumores benignos. Adicionalmente, observou-se uma forte correlação negativa entre a expressão de vimentina em células de reserva e expressão de pancitoqueratina em células diferenciadas (=-0,65, p<0,0001), além de maiores taxas de proliferação celular (Ki-67) e contagem de mitoses em tumores malignos em comparação com tumores benignos. Em análise univariada de sobrevida doença-específica, os fatores prognósticos foram a presença de invasão vascular ou metástase no momento do diagnóstico, o escore de E-caderina em células de reserva e diferenciadas, e o tamanho tumoral. Em relação ao tempo de sobrevida livre de doença, os fatores prognósticos foram a presença de invasão vascular ou metástase no momento do diagnóstico, escore de E-caderina em células de reserva e diferenciadas, perda de polaridade celular, grau de infiltração, histórico de doença recorrente, escore de morfologia fusiforme e proporção de células de reserva. Os resultados do presente estudo confirmaram que existe um grau de discordância diagnóstica entre patologistas que poderia explicar parte da variação observada na prevalência de carcinomas de glândula hepatoide em literatura, e de que existe potencial para melhora da concordância diagnóstica por meio dos algoritmos propostos. Ainda, os achados reforçaram a hipótese de que o mecanismo de transição epitélio-mesenquimal participa na progressão tumoral em neoplasias hepatoides em cães, explicando a frequente imunoexpressão de vimentina observada nestes tumores. Por fim, foi demonstrado que a presença de invasão vascular ou metástase no momento do diagnóstico tem impacto significativo na sobrevida dos animais, assim como há outras características clínicas, morfológicas e imuno-histoquímicas com valor prognóstico para recorrência da doença e sobrevida.

An important variation in the prevalence of canine hepatoid gland carcinoma is currently reported, likely due to lack of consistent and well-defined criteria. Additionally, clinical and pathological features predictive of biological behavior are not well defined. The epithelial-mesenchymal transition mechanism has been proposed as a possible explanation for the frequent expression of vimentin by neoplastic hepatoid gland cells, but this hypothesis remains elusive. The present study aimed to define the level of diagnostic agreement between different pathologists, investigate immunohistochemical and morphological features indicative of epithelial-mesenchymal transition, and to evaluate candidate features that might be predictive of biological behavior. Five pathologists diagnosed and classified morphological features for 57 hepatoid gland tumors, and diagnostic algorithms were built with recursive partitioning analysis. Additionally, vimentin, pancytokeratin, E-cadherin and ki-67 immunohistochemical expression were evaluated for 78 lesions, and expression levels were compared between normal glands, benign tumors, malignant tumors, and vasculo-invasive/metastatic tumors. Local recurrence free survival, disease free survival, overall survival and disease-specific survival were evaluated for 33 cases. There was a fair mean interobserver agreement for diagnosis (=0,54), that could potentially be improved to a good mean agreement with the use of two novel diagnostic algorithms (=0,64 and 0,65). Additionally, higher levels of vimentin expression and lower levels of E-cadherin expression were noted in malignant tumors, as compared to benign tumors, and there was a strong negative correlation between vimentin expression in reserve cells and pancytokeratin expression in differentiated cells (=-0,65, p<0,0001). Proliferation index (ki67) and mitotic count were higher for malignant tumors when compared to benign tumors. In univariate survival analysis, vascular invasion or metastasis at the time of diagnosis, E-cadherin score in differentiated and reserve cells and tumor size were predictive of disease-specific survival. Vascular invasion or metastasis at the time of diagnosis, E-cadherin score in differentiated and reserve cells, loss of cell polarity, grade of invasiveness, history of recurrent disease, spindle cell morphology score and proportion of reserve cells were predictive of disease-free survival. The results of this study demonstrate a level of diagnostic disagreement between pathologists that might explain, at least partially, the variation of prevalence of hepatoid gland carcinoma in current literature, and that this diagnostic agreement could potentially be improved. Also, these findings support the role of epithelial-mesenchymal transition in the progression of canine hepatoid gland tumors, and this mechanism could explain the frequent expression of vimentin in this particular neoplasia. Finally, it was shown that vascular invasion or metastasis at the time of diagnosis has a significant impact on survival, and that additional clinical, morphological and immunohistochemical features might also have predictive value for disease recurrence and survival.

EFEITOS DA FUMONISINA B1, DESOXINIVALENOL E ÁCIDO FÍTICO SOBRE EXPLANTES JEJUNAIS DE SUÍNOS: avaliação morfológica, imuno-histoquímica, expressão de citocinas e estresse oxidativo

O objetivo do presente estudo foi avaliar os efeitos tóxicos das micotoxinas desoxinivalenol (DON) e fumonisina B1 (FB1) sobre o intestino e investigar a ação moduladora do ácido fítico (IP6) por meio de um modelo ex vivo, analisando a morfologia, proliferação celular, apoptose, expressão de E-caderina e ciclooxigenase-2 (cox-2), estresse oxidativo e expressão de citocinas em jejuno de suínos. Para tal, no primeiro experimento, foram utilizados cinco suínos com 24 dias de idade, os quais foram eutanasiados, o jejuno foi excisado, os explantes colhidos (40 explantes/animal) com um punch de 8 mm de diâmetro e submetidos a oito tratamentos (seis explantes/tratamento) : controle (somente meio de cultura), IP6 5mM, FB1 70µM, DON 10 µM, DON 10 µM+ FB1 70 µM, DON 10µM + IP6 5 mM, FB1 70 µM+IP6 5 mM e DON 10 µM+ FB1 70 µM+IP6 5 mM, incubados durante quatro horas sob agitação constante em estufa a 37°C. Após esse período, três explantes de cada tratamento foram fixados em solução de formalina tamponada a 10% e processados para realização do exame histológico e imuno-histoquímico para os anticorpos anti-ki-67, anti-caspase-3, anti-E-caderina e anti-cicloxigenase-2. Para a análise do estresse oxidativo, três explantes foram congelados em nitrogênio líquido, armazenados a -80C° e posteriormente processados para a avaliação dos níveis de glutationa reduzida (GSH), substâncias reativas ao ácido tiobarbitúrico (TBARS) e capacidade da atividade antioxidante por meio dos testes de 2, 2azinobis-3-etilbenzotiazolina-6-ácido sulfônico (ABTS) e poder de redução do ferro (FRAP). Alterações morfológicas, diminuição da expressão de E-caderina, aumento da proliferação celular, apoptose, expressão de cox-2 e ocorrência de estresse oxidativo foram mais evidentes nos explantes expostos aos tratamentos com DON. A presença de IP6 reduziu significativamente as alterações morfológicas, proliferação celular, apoptose, expressão de cox-2 e estresse oxidativo, e aumentou a expressão de E-caderina nos explantes jejunais expostos às micotoxinas. Para avaliação da expressão de citocinas e peptídeos antimicrobianos, um segundo experimento com metodologia similar ao primeiro foi realizado. Explantes jejunais foram colhidos (21 explantes/animal) e submetidos aos seguintes tratamentos (três explantes/tratamento): controle (somente meio de cultura), FB1 70µM, FB1 70 µM+IP6 2,5 mM, FB1 70 µM+IP6 5 mM, DON 10 µM, DON 10µM + IP6 2,5 mM e DON 10µM + IP6 5 mM. Após a incubação, os explantes foram congelados em nitrogênio liquido e armazenados a - 80°C para avaliação da expressão das citocinas pró-inflamatórias (IL-1, IL-6, IL-8, IL-10, IFN-, TNF-) e anti-inflamatórias (IL-10) e -defensinas 1 (pBD-1) e 2 (pBD-2) por meio da reação em cadeia pela polimerase em tempo real. A exposição às micotoxinas induziu a alterações na expressão das citocinas e -defensinas. A presença de IP6, principalmente na dose de 5mM modulou os efeitos das micotoxinas, tornando a expressão da resposta inflamatória e antimicrobiana similar aos explantes controle. Os resultados do presente estudo evidenciaram os efeitos tóxicos da FB1 e DON sobre o jejuno de suínos, assim como o efeito protetor do IP6 em modular as alterações morfológicas, imunológicas e do estresse oxidativo induzido por tais micotoxinas.

The aim of the present study was to evaluate the toxic effects of mycotoxins fumonisin B1 (FB1) and deoxynivalenol (DON) on the intestine, and investigate the modulatory action of phytic acid (IP6) through of ex vivo model and assessment of morphology, cell proliferation, apoptosis, E-cadherin and cyclooxygenase-2 (cox-2) expression, oxidative stress and cytokines and antimicrobial peptides in jejune of swine. In the first experiment, five pigs 24-days-old were euthanized and the jejunal explants were collected (40 explants/animal) using a punch of 8 mm in diameter and submitted to eight treatments (six explants/treatment): control (culture medium), IP6 5mM, FB1 70µM, DON 10 µM, DON 10 µM+ FB1 70 µM, DON 10µM + IP6 5 mM, FB1 70 µM+IP6 5 mM and DON 10 µM+ FB1 70 µM+IP6 5 mM, and incubated at 37°C under orbital shaking. After the incubation period, three explants per treatment were fixed in 10% neutral buffered formalin and processed for the histological examination and immunohisotchemistry assay using antibodies against ki-67, cleaved caspase-3, e-cadherin and cox-2. Three explants were immediately frozen in liquid nitrogen and posteriorly stored at -80°C to the assessment of reduced glutathione (GSH), thiobarbituric acid reactive substances (TBARS), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) and ferric reducing antioxidant power (FRAP) levels. Morphological changes, decrease of E-cadherin expression and increase of cell proliferation, apoptosis, cox-2 expression and oxidative stress were more evident in the explants exposed to DON treatments. The presence of IP6 significantly decreased the morphological changes, cell proliferation, apoptosis, cox-2 expression and oxidative stress, and increased the E-cadherin expression in the jejunal explants exposed to mycotoxins in all treatments. A second experiment with similar methodology was performed to evaluate the cytokines and antimicrobial peptides expression. The jejunal explants without serosa were collected (21 explants/animal) and submitted to treatments (three explants/treatment): control (medium culture), FB1 70µM, FB1 70 µM+IP6 2.5 mM, FB1 70 µM+IP6 5 mM, DON 10 µM, DON 10µM + IP6 2.5 mM and DON 10µM + IP6 5 mM. After incubation, the explants were frozen in liquid nitrogen and posteriorly stored at -80°C to the assessment of pro- (IL-1, IL-6, IL-8, IL-10, IFN-, TNF-) and anti-inflammatory (IL-10), and -defensins 1 (pBD-1) and 2 (pBD-2) by the reverse transcription polymerase chain reaction quantitative real time assay. The exposition to mycotoxins induced to changes in the cytokines and -defensins expression. The presence of IP6, mainly in the 5 mM concentration, modulated the mycotoxins effects, becoming the expression of inflammatory and antimicrobial response similar to the control explants. The results of present study evidenced the toxic effects of FB1 and DON on the swine jejune, and the protective effect of IP6 in modulate the morphological and immunological changes, and the oxidative stress induced by these mycotoxins.