Ki-67 labeling in canine perianal glands neoplasms: a novel approach for immunohistological diagnostic and prognostic

Background: The antibody Ki-67 is a reliable and easy tool to accurately assess the growth fraction of neoplasms in humans and animals, and it has been used to predict the clinical outcome. Therefore, the aim of the present study was to investigate the immunohistochemical expression pattern of Ki-67 in normal and neoplastic perianal glands of dogs to evaluate the possible use of this proliferation marker as an ancillary method of perianal tumor diagnosis. We studied 42 cases of perianal gland neoplasms including adenomas (n = 15), epitheliomas (n = 15), and carcinomas (n = 12). As controls, 13 tissue samples from normal perianal glands were used. A Ki-67 index was established by a computer-assisted image analysis and compared with manual counting. Results: Out of the 42 cases of perianal gland neoplasms, 34 were from males and eight from females. Recurrence was reported in 14 cases, being higher (8/12) in carcinomas. Immunostaining for Ki-67 revealed that the carcinomas showed a higher proliferation rate (9.87%) compared to groups of epitheliomas (2.66%) and adenomas (0.36%). For adenomas and epitheliomas of the perianal glands the computer-assisted counting and the manual counting gave similar results; however, only the computer-assisted image analysis was efficient to predict the perianal gland carcinoma recurrence.Conclusion: Since there were significant differences in the number of Ki-67-positive nuclei, this marker proved to be effective in helping the classification of perianal gland neoplasms and to refine the diagnosis criteria, especially in those samples with high variation in morphology/area. Also, higher Ki-67 index is related to recurrence in cases of perianal gland carcinomas. Further, the computer-assisted image analysis proved to be a fast and reliable method to assess the Ki-67 index in perianal gland neoplasms. © 2013 Pereira et al.; licensee BioMed Central Ltd

Imuno-histoquímica na patologia veterinária diagnóstica: uma revisão crítica

Immunohistochemistry has become a practical and widely used tool for diagnosis in human pathology since the 70's. However, its application in veterinary diagnostic pathology has not been so common, especially due to the lack of specific antibodies. To overcome this drawback, antibodies which present cross reactivity with human and animal antigens have been applied. The purpose of the present study was to test the cross reactivity of some antibodies intended for the human pathology, which may be used in animal tissues, with the help of antigen retrieval and amplification systems. In the present study it was confirmed that many of the antibodies produced for use in human histopathology might be applied in veterinary pathology. Further studies are needed to increase the list of applicability of these antibodies to different animal species. It must be stressed that in this type of study some variables, such as clone of antibody, dilution, antigen retrieval method, and detection system, have to be evaluated.A técnica de imuno-histoquímica é usada na rotina diagnóstica e na pesquisa em patologia humana desde 1970, porém seu uso na patologia veterinária é relativamente recente, principalmente com objetivo diagnóstico. A maior dificuldade no uso da imuno-histoquímica na patologia veterinária tem sido a falta de anticorpos específicos para os tecidos animais. Na falta de anticorpos específicos para as espécies domésticas, a patologia veterinária freqüentemente faz uso de anticorpos que apresentam reatividade cruzada entre antígenos humanos e animais. O objetivo deste trabalho foi testar a reatividade cruzada de diversos anticorpos feitos para uso humano em tecido parafinado de algumas espécies animais, utilizando-se dos novos métodos de recuperação antigênica e amplificação da reação imuno-histoquímica. No presente estudo foi possível confirmar a aplicabilidade de que muitos anticorpos produzidos para diagnóstico imuno-histoquímico em patologia humana podem ser utilizados em patologia veterinária. Novos estudos são necessários a fim de se ampliar a lista de aplicabilidade desses anticorpos em diferentes espécies animais, levando sempre em consideração as variações de clones, diluições, métodos de recuperação antigênica e de revelação.26327

Concentração de proteínas de fase aguda e vitamina D em cães com linfoma multicêntrico

Objetivou-se caracterizar a concentração sérica da vitamina D e das PFAs (Proteínas de Fase Aguda) (alfa-1 glicoproteína ácida, haptoglobina, transferrina, ceruloplasmina, albumina, IgA, IgG e alfa-1 – antitripsina) em cães com linfoma multicêntrico, submetidos ao tratamento quimioterápico com protocolo CHOP (Ciclofosfamida, Doxorrubicina, Vincristina e Prednisona), determinando o valor prognóstico desses marcadores para a doença. Foram avaliadas as concentrações séricas das PFAs, através do método da eletroforese e as concentrações da vitamina D, através da quimioluminescência em dois grupos experimentais, um grupo de 13 cães com linfoma multicêntrico classificados como alto grau pela citologia (GL) durante as semanas T0, T5 e T10 do tratamento com protocolo quimioterápico antineoplásico e em um grupo de 10 animais saudáveis para compor o grupo controle (GC), em coleta única. Para isso, foi realizado o diagnóstico, estadiamento e avaliação de resposta terapêutica dos 13 pacientes com linfoma multicêntrico através de técnicas de citopatologia, histopatologia, imuno-histoquímica do linfonodo periférico acometido. Foi observado que 9 pacientes tiveram resposta completa e 4 pacientes tiveram resposta parcial ao tratamento. Os dados foram analisados através do software R. Os resultados indicam que as diferenças entre as variáveis IgA, haptoglobina e α1-glicoproteína ácida foram significativas entre os grupos, e entre os diferentes momentos da quimioterapia (p< 0,05), indicando que podem ser marcadores sensíveis ao linfoma em cães. A α1-glicoproteína ácida apresentou valor prognóstico para o linfoma, com 63% de especificidade. Porém a vitamina D não apresentou valor prognóstico para o linfoma multicêntrico em cães. Palavras-chave: linfossarcoma; prognóstico; reação de fase aguda; resposta terapêutica; neoplasia hematopoiética

Histomorphological and immunophenotypic diagnoses of gastrointestinal stromal tumors and other sarcomas that affect the intestine of dogs

In view of the morphological similarity between gastrointestinal stromal tumors (GIST) and other sarcomas of the intestine of dogs, the aim was to carry out the histomorphological and immunohistochemical diagnosis of these tumors, associating breed, sex and age, location and tumor invasion. 217 cases were evaluated by histopathology and 36 diagnosed by immunohistochemistry were included (24 GIST and 12 other intestinal sarcomas). Mixed breed dogs were the most diagnosed with GIST, mainly elderly females (9.5±2.2 years); in the other intestinal sarcomas, crossbreeds and Dachshunds, males and females, were equally affected. The cecum was the most affected by GISTs, with tumor invasion of the intestinal layers in all cases. The small intestine was the most affected by the other intestinal sarcomas, with invasion of the layers in most of these tumors. GISTs expressed markers such as CD117 and DOG-1, unlike other intestinal sarcomas. GIST and other intestinal sarcomas denoted histomorphological and immunophenotypic characteristics similar to histopathology, justifying the association of immunohistochemistry for the definitive diagnosis. Keywords: CD117; DOG-1; GIST; veterinary oncolog

Diagnosis, Prognosis and Treatment of Canine Cutaneous and Subcutaneous Mast Cell Tumors

Mast cell tumors (MCTs) are hematopoietic neoplasms composed of mast cells. It is highly common in dogs and is extremely important in the veterinary oncology field. It represents the third most common tumor subtype, and is the most common malignant skin tumor in dogs, corresponding to 11% of skin cancer cases. The objective of this critical review was to present the report of the 2nd Consensus meeting on the Diagnosis, Prognosis, and Treatment of Canine Cutaneous and Subcutaneous Mast Cell Tumors, which was organized by the Brazilian Association of Veterinary Oncology (ABROVET) in August 2021. The most recent information on cutaneous and subcutaneous mast cell tumors in dogs is presented and discussed

Immunohistochemistry in diagnostic veterinary pathology: a critical review

A técnica de imuno-histoquímica é usada na rotina diagnóstica e na pesquisa em patologia humana desde 1970, porém seu uso na patologia veterinária é relativamente recente, principalmente com objetivo diagnóstico. A maior dificuldade no uso da imuno-histoquímica na patologia veterinária tem sido a falta de anticorpos específicos para os tecidos animais. Na falta de anticorpos específicos para as espécies domésticas, a patologia veterinária freqüentemente faz uso de anticorpos que apresentam reatividade cruzada entre antígenos humanos e animais. O objetivo deste trabalho foi testar a reatividade cruzada de diversos anticorpos feitos para uso humano em tecido parafinado de algumas espécies animais, utilizando-se dos novos métodos de recuperação antigênica e amplificação da reação imuno-histoquímica. No presente estudo foi possível confirmar a aplicabilidade de que muitos anticorpos produzidos para diagnóstico imuno-histoquímico em patologia humana podem ser utilizados em patologia veterinária. Novos estudos são necessários a fim de se ampliar a lista de aplicabilidade desses anticorpos em diferentes espécies animais, levando sempre em consideração as variações de clones, diluições, métodos de recuperação antigênica e de revelação.Immunohistochemistry has become a practical and widely used tool for diagnosis in human pathology since the 70's. However, its application in veterinary diagnostic pathology has not been so common, especially due to the lack of specific antibodies. To overcome this drawback, antibodies which present cross reactivity with human and animal antigens have been applied. The purpose of the present study was to test the cross reactivity of some antibodies intended for the human pathology, which may be used in animal tissues, with the help of antigen retrieval and amplification systems. In the present study it was confirmed that many of the antibodies produced for use in human histopathology might be applied in veterinary pathology. Further studies are needed to increase the list of applicability of these antibodies to different animal species. It must be stressed that in this type of study some variables, such as clone of antibody, dilution, antigen retrieval method, and detection system, have to be evaluated

Avaliação da caspase-3 e Ki-67 como marcadores prognósticos nas neoplasias mamárias em cadelas

Apoptosis, as a cellular event, has an important participation in tumorigenesis, determining the tumoral growth and aggressiveness. The present study had as objective to evaluate the occurrence of apoptosis, associated cellular proliferation, in canine mammary neoplasias and the patient clinical evolution. Seventy dogs had been submitted to surgical excision of the tumor fragment and they were submitted to the histopathologic diagnosis and imunohistochemistry procedure to caspase-3 and the Ki-67. This apoptosis and cellular proliferation markers demonstrated great expression in malignant neoplasias especially carcinoma, considered the most malignant of the tumors. These results confirm consulted literature contributing for a criterious tumoral prognostic complementing the tumoral classification, providing a greater and better supervened due to adequacy of therapeutic procedure of each patient.A apoptose e a proliferação celular possuem uma participação importante na tumorigênese, determinando o crescimento tumoral e consequentemente sua agressividade. O presente estudo teve como objetivo avaliar a ocorrência da apoptose associada a proliferação celular em neoplasias mamárias em cadelas e a elas a evolução clínica do paciente. Setenta animais foram submetidos à exérese cirúrgica do tumor, sendo este submetido ao diagnóstico histopatológico e marcação imuno-histoquímica para caspase-3 e Ki-67. Estes marcadores de apoptose e proliferação celular demonstraram grande expressão nas neoplasias malignas, principalmente nos carcinomas, considerado o mais maligno dos tumores. Estes resultados corroboram os dados da literatura e contribuem para um prognóstico tumoral criterioso que complementa a classificação tumoral existente proporcionando uma melhor e maior sobrevida devido a uma adequação do procedimento terapêutico de cada paciente

Investigation of Prognostic Value of Claudin-5, PSMA, and Ki67 Expression in Canine Splenic Hemangiosarcoma

Splenic hemangiosarcoma (HSA) is a malignant tumor of endothelial cells that affects middle-aged and elderly dogs and is characterized by the formation of new blood vessels, commonly associated with necrotic and hemorrhagic areas. Despite its importance in veterinary medicine, few studies have identified markers with prognostic value for canine HSA. Thus, this study aimed to associate the clinicopathological findings (prostate-specific membrane antigen [PSMA], Claudin-5, and Ki67 gene and protein expression) with overall survival in HSA-affected patients. Fifty-three formalin-fixed and paraffin-embedded canine splenic HSA samples, previously diagnosed by histopathological examination, were used in this study. Claudin-5, PSMA, and Ki67 protein expression levels were evaluated by immunohistochemistry, and gene expression was evaluated by quantitative polymerase chain reaction. Claudin-5 protein overexpression was observed in patients with metastasis (p = 0.0078) and with stage III tumors compared to those with stage I and II tumors (p = 0.0451). In patients treated with surgery alone, low PSMA gene and protein expression (p = 0.05 and p = 0.0355, respectively) were associated with longer survival time. Longer survival time was observed in patients with a low Ki67 index (p = 0.0488). Our results indicate that Claudin-5 protein expression is associated with metastatic status, and PSMA gene and protein expression, and Ki67 index are associated with survival time