Expression of LIN28A/B and Let-7 miRNAs in canine mammary carcinomas / Expressão de LIN28A/B e Let-7 miRNAs em carcinomas mamários caninos

LIN28 is a RNA-binding protein including two highly conserved homologous, LIN28A and LIN28B. Proto-oncogenes such as LIN28A and LIN28B are generally targeted by the let-7 miRNAs in different types of human cancers. Here, we determined the expression of LIN28A in canine mammary tumor samples and the LIN28/let-7 pathway in canine mammary cell lines. In those cell lines, we identified a functional LIN28/let-7 pathway which exhibited high expression of let-7 members and low expression of its targets, including LIN28A and LIN28B. However, the mammary carcinoma tissue samples showed a frequent expression of LIN28A being expressed mainly in the epithelial cells. No association was observed between LIN28A expression and histopathological classification and grade, TNM and survival time. Our results suggested a possible role of the LIN28A protein in the development of canine mammary carcinomas due to the high frequency observed in the tumor samples (28 of 32). The in vitro experiments suggested that the LIN28/let-7 pathway is active in the tumor cells evaluated. However, more studies are necessary to elucidate the exact role of LIN28/let-7 pathway in canine mammary carcinomas.

LIN28 é uma proteína de ligação ao RNA, com duas formas homólogas altamente conservadas, LIN28A e LIN28B. Os proto-oncogenes LIN28A e LIN28B são regulados pela família de miRNAs let-7 em diferentes tipos de cânceres em humanos. No presente trabalho, o objetivo foi determinar a expressão de LIN28A em amostras de tumor mamário de cadelas e a via LIN28/let-7 em linhagens celulares mamárias caninas. Nestas linhagens, através das técnicas de qPCR e RNAseq, foi identificado que a via LIN28/let-7 apresenta-se funcional, com alta expressão dos membros da família let-7 e baixa expressão de seus alvos, entre eles LIN28A e LIN28B. No entanto, as amostras de tecidos de carcinomas mamários caninos demonstraram expressão frequente de LIN28A, sendo observada principalmente em células epiteliais. Não foram observadas associações entre expressão de LIN28A com classificação e gradação histopatológicas, TNM e tempo de sobrevida. Nossos resultados sugerem uma possível relação da proteína LIN28A no desenvolvimento de carcinomas mamários caninos devido à alta frequência observada nas amostras tumorais (28 de 32). Os experimentos in vitro sugerem que a via LIN28/let-7 é ativa nas linhagens celulares caninas avaliadas. Entretanto, estudos funcionais ainda são necessários para elucidar a função exata da via LIN28/let-7 nos carcinomas mamários caninos.

Exérese tumoral seguida de blefaroplastia no tratamento de carcinoma de células escamosas em pálpebra inferior de equino / Tumoral resection followed by blepharoplasty in squamous cell carcinoma treatment in the lower eyelid of a horse

Background: Equines are routinely subjected to enucleation due to palpebral tumors. Blepharoplasties in horses, especially in the lower eyelid, are rarely performed due to the difficulty of sliding once the tissue around the eyes presents low mobility. Defects involving more than 50% of the lower eyelid is considered challenging after tumor removal. Squamous cell carcinoma (SCC) is the second most reported neoplasm in horses, being very common in regions of the lower eyelid, third eyelid, sclera and or cornea. The aim of this study is to present the Destro VY skin advancement flap as a blepharoplasty technique performed after surgical excision of a SCC, with total commitment of the lower eyelid, completely covering the right eye of a mare. Case: A 8-year-old mare Paint Horse, weighing 420 kg, was referred for evaluation of tumor tissue of 6.0 x 4.0 x 2.0 cm, with nodular and ulcerated appearance, involving the right lower eyelid, in its total extension and completely covering the eye, without adhering to it. Considering the initial suspicion of SCC, the treatment strategy performed was surgical eyelid excision and maintenance of the eye, followed by blepharoplasty as an attempt to reconstruct the eyelid. Under general inhalation anesthesia, the animal was placed in left lateral recumbency, when the surgical region was prepared and local anesthetic block was performed. After antisepsis, a skin incision was made circumscribing the tumor, respecting a margin of 10 mm apart and excision of all visible tumor tissue was performed followed by intralesional ozone therapy. Blepharoplasty was performed to cover the portions of the exposed lacrimal and zygomatic bones, as well as correction of the eyelid aesthetics. For this, Destro VY skin advancement flap was performed for reconstruction of the lower eyelid. An incision of approximately 7 cm in Vshaped skin was performed, and the subcutaneous tissue under the V was dissected, maintaining a central pedicle, responsible for the vascularization of the flap, which was slid, approximately 20 mm, towards the eye. After obtaining the desired skin approximation, Y-suture was performed, covering the exposed bone and reconstructing the lower eyelid. In the postoperative period, local instillation of mitomycin eye drops and systemic meloxicam administration were instituted. The mare had her vision restored, presenting satisfactory morpho functional and aesthetic results and no tumor recurrence during 1-year of follow-up. Discussion: The repair of lower eyelid imperfections is challenging, especially when they have large defects, and there are no reports of performing the Destro VY skin advancement flap technique in horses for lower eyelid reconstruction. In this case, the importance of the blepharoplasty technique is emphasized, avoiding enucleation, preserving horse's vision and aesthetics. In addition, aiming to avoid tumor recurrence, especially if surgical safety margins can not be achieved, other complementary treatments should be associated, including intralesional ozone therapy, mitomycin, an antineoplastic chemotherapy drug, and meloxicam, a COX-2 selective, non-steroidal anti-inflammatory drug, as performed in this study. It is concluded that the use of the Destro VY skin advancement flap technique for reconstruction of the external lamella in cases of SCC in the lower eyelid of horses is a feasible technique, which preserves the animal's vision, as well as aesthetics. The safety margin in the surgical excision of the SCC and the association of complementary therapies in the resolution of the condition are important points also to be considered.

Integration of heterologous bone matrix associated with polymethylmethacrylate in induced tibial bone defects. An experimental study in rabbits

ABSTRACT Purpose To analyze and compare the reactions at the interface between the composite, composed of fragmented heterologous mineralized bone matrix (MOMHF) and polymethylmethacrylate (PMMA), and the rabbit's tibias, through macroscopic evaluation and scanning electron microscopy (SEM) in different periods. Methods In this study, 12 New Zealand adult rabbits were used (E1: n = 3, E2: n = 3, E3: n = 3 and E4: n = 3). They had the right tibial defects filled with composite and were evaluated immediately after surgery and at 30, 60, 90, and 120 days. Results The composites were incorporated and integrated into the recipient beds in 100% of the cases, defined by the MOMHF osseointegration and the PMMA fibrointegration, with no sign of infection, migration, or rejection. Conclusions The behavior of the composites in the recipient beds demonstrates that these biomaterials have the potential to be used in bone defect repairs, offering, thus, better quality of life to the orthopedic patient.

Clinical and complementary diagnosis of sinus neoplasia in horses

Background: Sinus neoplasms are reported as low frequency in horses. Its clinical characteristics are often nonspecific, depend on complementary methods for diagnosis, and when diagnosed, generally they are already advanced, limiting therapeutic possibilities. The objective of this case series was to detail clinical aspects and complementary exams for sinus neoplasms for early diagnosis, comparing them with the literature. Cases: Four horses were treated at the College of Veterinary Medicine and Animal Science of the University of São Paulo, with different clinical signs and previous diagnoses, which when evaluated by respiratory endoscopy, radiography, oral cavity evaluation and histopathological exams, revealed the diagnosis of anaplastic carcinoma, poorly differentiated carcinoma, ossifying fibroma and lymphoma. Previous diagnosis, based mainly on clinical signs, were sinusitis secondary to apical infection, ethmoidal hematoma, sinus cyst and sinusitis secondary to periapical disease. The cases of anaplastic carcinoma (case 1) and lymphoma (case 4) presented with an advanced degree of the disease with involvement and destruction of paranasal structures and adjacent tissues, in addition to a poor general condition, which made surgical treatment impossible and led to euthanasia. In these cases, previous treatment was made to apical infection and periodontal disease with secondary sinusitis, but negative evolution led to suspicion of neoplasia, confirmed by histopathological exam of incisional biopsy of the mass in oral cavity. Benign ossifying fibroma (case 3) presented with progressive respiratory difficulty due to occlusion of the lumen of some nasal meatus and radiographic signs of invasion and deformation of the adjacent bones, it was submitted for surgical excision and there was no recurrence until hospital discharge. The poorly differentiated carcinoma (case 2) was a multilobulate neoformation in the ethmoidal region, similar to ethmoid hematoma in endoscopic and radiographic evaluation, it was submitted for excision and two sessions of electrochemotherapy with bleomycin, associated with administration of piroxicam, which obtained a good result until the period of 1 year after discharge. Discussion: It was found that many characteristics are common with these types of neoplasms, and the clinical signs, such as nasal secretions, airway obstruction, increased facial volume, severe alterations in oral cavity, although unspecific, suggest the differential diagnosis for neoplasms. It is important to differentiate from other diseases noting the evolution and growth of these tumors, like in cases 1 and 4, especially the growth to internal tissues, using complementary methods described here, like endoscopic and radiographic examination. Late identification or even manipulation of neoplasms, without proper diagnosis, leads to a few prognoses regarding life. When it is possible to perform surgical excision, complementary methods are important to guide the procedure, and definitive diagnosis is made through histopathologic exam and some need immunohistochemistry analysis. Cases 2 and 3 had surgical access, were submitted to excision and treatment with good results, both with close monitoring in first months and prolonged quality of life. These results highlight the importance of complementary methods for early diagnosis, correct intervention and monitoring of evolution.

Comparative morphometric evaluation of hepatic hemosiderosis in wild Magellanic penguins (Spheniscus magellanicus) infected with different Plasmodium spp. subgenera / Avaliação morfométrica comparativa da hemossiderose hepática em pinguins-de-Magalhães (Spheniscus magellanicus) de vida livre infectados por diferentes subgêneros de Plasmodium spp

Avian malaria is one of the most important diseases of captive penguins. We employed morphometric techniques to evaluate hepatic hemosiderosis in rehabilitating wild Magellanic penguins (Spheniscus magellanicus) that were negative (n = 9) or naturally infected by different subgenera of Plasmodium spp. (n = 24), according with: Plasmodium subgenera (Haemamoeba, Huffia, Other lineages, and Unidentified lineages), severity of Plasmodium histopathological lesions, and concurrent diseases, age class (juvenile or adult plumage), sex (male, female or not determined), body score (emaciated, thin, good, excellent, not available), molt, presence or absence of oil contamination upon admission, iron supplementation, and rehabilitation center. The percentage of the area occupied by hemosiderin was called Index of Hepatic Hemosiderosis (IHH). Plasmodium-positive females presented significantly higher IHH values (17.53 ± 12.95%) than males (7.20 ± 4.25%; p = 0.041). We observed higher levels of congestion (p = 0.0182) and pneumonia (p = 0.0250) severity between Unidentified lineages vs. Huffia. We believe that the hepatic hemosiderosis observed in this study was multifactorial, the result of pathological processes caused by malaria, molting, hemoglobin and myoglobin catabolism during migration, anemia, concomitant diseases, and iron supplementation, all possibly potentiated by decreased liver mass. Further studies are needed to clarify the mechanisms of these hypotheses.(AU)

Malária aviária é uma das mais relevantes doenças em pinguins cativos. Foram aplicadas técnicas morfométricas para avaliar a hemossiderose hepática em pinguins-de-Magalhães (Spheniscus magellanicus ) de vida livre em reabilitação negativos (n = 9) e naturalmente infectados por diferentes subgêneros de Plasmodium spp. (n = 24), quanto a: subgênero de Plasmodium (Haemamoeba , Huffia, Outras Linhagens, e Linhagens não identificadas), severidade das lesões histopatológicas causadas por Plasmodium e doenças concomitantes, faixa etária (plumagem juvenil ou adulta), sexo (macho, fêmea, indeterminado), condição corporal (emaciado, magro, bom, excelente, indisponível), muda, presença/ausência de óleo a admissão, suplementação de ferro, e centro de reabilitação. A porcentagem da área ocupada por hemossiderina foi denominada Índice de Hemossiderose Hepática (IHH). Fêmeas Plasmodium -positivas apresentaram IHH significativamente mais elevado que machos, respectivamente, 17,53 ± 12,95% e 7,20 ± 4,25% (p = 0,041). Níveis mais elevados de congestão (p = 0,0182) e pneumonia (p = 0,0250) foram observados entre Linhagens não identificadas vs. Huffia. Possivelmente, a hemossiderose hepática observada nesse estudo seja multifatorial, resultado de processos patológicos causados por malária, muda, catabolismo de hemoglobina e mioglobina durante a migração, anemia, doenças concomitantes e suplementação de ferro, potencialmente intensificados por massa hepática reduzida. Estudos complementares são necessários para esclarecer os mecanismos de tais hipóteses.(AU)

Evaluation of polyamide surgical mesh as an abdominal ventral implant in rabbits

Purpose: To evaluate abdominal ventral wound healing by using a specific biomaterial, a handmade polyamide surgical mesh. Methods: A surgical incisional defect was made in ten rabbits to simulate a hernia in the ventral abdominal musculature. A polyamide surgical mesh was used in hernioplasty. They were monitored for surgical wound healing, and macroscopically and histologically evaluated at the end of the experiment. The polyamide surgical mesh did not cause foreign body reaction, pain, edema, or infection in the surgical site. The manure production was not affected by intestinal tissue adherences to the mesh, consistent with the ultrasonography result where adherences were not observed and organized scarring tissue formed in the incisional defect. The polyamide mesh was fixed over the abdominal wall, and its external and internal sides were surrounded by a vascularized connective tissue. Results: None of the experimental animals developed adherences from internal organs to the polyamide mesh, except two rabbits where the omentum formed adherence to the internal scarring tissue without present herniation or compromise of the rabbits health. Conclusion: Polyamide surgical mesh for hernioplasty presents, in rabbits, excellent biocompatibility, with minimal body adverse reactions and low cost.(AU)

Comparative leukocyte morphometric analysis between endemic Anurans from Brazil and the invasive species Lithobates catesbeianus / Análise morfométrica comparada entre Anuros endêmicos do Brasil e a espécie invasora Lithobates catesbeianus

Amphibians are potentially reliable and efficient bioindicators. Existing anuran white blood cell morphology studies are limited, with only a few morphometric studies available. We employed morphometric techniques to characterize leukocytes of selected Neotropical anurans from Brazil and compare our findings with the exotic American Bullfrog (Lithobates catesbeianus), genus Ranidae. We compared blood smears of 28 specimens from six different genera (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus, and Proceratophrys) with samples from L. catesbeianus. Leukocyte average diameter was calculated by an image analysis software. One-way analyses of variance and Bonferroni tests were used on statistical analyses. Lymphocytes, neutrophils, eosinophils, and basophils were significantly smaller than the reference ranges reported for other amphibian genera, including Lithobathes, whereas monocyte diameters did not differ significantly between genera. This is the first study to evaluate leukocyte morphometrics of Brazilian anuran species. Our findings suggest that geographical separation could possibly influence leukocyte morphometry.(AU)

Anfíbios são indicadores ambientais potencialmente confiáveis e eficientes. Estudos referentes à morfologia de leucócitos de anuros são limitados, com poucos estudos morfométricos disponíveis em literatura. O presente estudo empregou técnicas morfométricas para caracterizar leucócitos de anuros Neotropicais brasileiros selecionados e compará-los com a espécie exótica rã-touro (Lithobates catesbeianus), família Ranidae. Esfregaços sanguíneos de 28 espécimes pertencentes a seis gêneros diferentes (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus e Proceratophrys) foram comparados com amostras de esfregaços de L. catesbeianus. A média do diâmetro dos leucócitos foi calculada por um software de análise de imagens. One-way e teste de Bonferroni foram utilizados para avaliação estatística. Linfócitos, neutrófilos, eosinófilos e basófilos mostraram-se significativamente menores que os valores de referência reportados em outros gêneros de anfíbios, incluindo Lithobathes; por outro lado, a média do diâmetro dos monócitos não demonstrou variação significativa entre os gêneros. Esse é o primeiro estudo de avaliação morfométrica de leucócitos em espécies de anuros brasileiros. Nossos resultados sugerem que a separação geográfica possivelmente influencia a morfometria leucocitária.(AU)

Comparative leukocyte morphometric analysis between endemic Anurans from Brazil and the invasive species Lithobates catesbeianus / Análise morfométrica comparada entre Anuros endêmicos do Brasil e a espécie invasora Lithobates catesbeianus

Amphibians are potentially reliable and efficient bioindicators. Existing anuran white blood cell morphology studies are limited, with only a few morphometric studies available. We employed morphometric techniques to characterize leukocytes of selected Neotropical anurans from Brazil and compare our findings with the exotic American Bullfrog (Lithobates catesbeianus), genus Ranidae. We compared blood smears of 28 specimens from six different genera (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus, and Proceratophrys) with samples from L. catesbeianus. Leukocyte average diameter was calculated by an image analysis software. One-way analyses of variance and Bonferroni tests were used on statistical analyses. Lymphocytes, neutrophils, eosinophils, and basophils were significantly smaller than the reference ranges reported for other amphibian genera, including Lithobathes, whereas monocyte diameters did not differ significantly between genera. This is the first study to evaluate leukocyte morphometrics of Brazilian anuran species. Our findings suggest that geographical separation could possibly influence leukocyte morphometry.(AU)

Anfíbios são indicadores ambientais potencialmente confiáveis e eficientes. Estudos referentes à morfologia de leucócitos de anuros são limitados, com poucos estudos morfométricos disponíveis em literatura. O presente estudo empregou técnicas morfométricas para caracterizar leucócitos de anuros Neotropicais brasileiros selecionados e compará-los com a espécie exótica rã-touro (Lithobates catesbeianus), família Ranidae. Esfregaços sanguíneos de 28 espécimes pertencentes a seis gêneros diferentes (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus e Proceratophrys) foram comparados com amostras de esfregaços de L. catesbeianus. A média do diâmetro dos leucócitos foi calculada por um software de análise de imagens. One-way e teste de Bonferroni foram utilizados para avaliação estatística. Linfócitos, neutrófilos, eosinófilos e basófilos mostraram-se significativamente menores que os valores de referência reportados em outros gêneros de anfíbios, incluindo Lithobathes; por outro lado, a média do diâmetro dos monócitos não demonstrou variação significativa entre os gêneros. Esse é o primeiro estudo de avaliação morfométrica de leucócitos em espécies de anuros brasileiros. Nossos resultados sugerem que a separação geográfica possivelmente influencia a morfometria leucocitária.(AU)

Occurrence of acid erosion of the teeth, actinomycosis and polioencephalomalacia in flock of sheep fed wet low pectin citrus pulp / Ocorrência de erosão ácida dos dentes, actinomicose e polioencefalomalacia em rebanho de ovinos alimentados com polpa cítrica úmida despectinada

The citrus pulp can be used as a substitute in ruminant feed reducing costs and maintaining the nutritional quality of food. However, this compound should be used carefully so as not to cause harm to the animals. The present report aims to describe the occurrence of dental erosion, actinomycosis and polioencephalomalacia in sheep raised and kept with a wet low pectin citrus pulp based diet, composing 50% of roughage. Actinomycosis was diagnosed in five animals through clinical and radiographic examinations and microbiological culture, and, after treatment, three animals were cured. Polioencephalomalacia was confirmed in ten animals by clinical diagnostics, in nine out of ten animals by therapeutic diagnosis, and in one animal by post-mortem anatomopathological examination. According to the observed, we recommend caution when large amounts of citrus pulp are used as bulky food.

A polpa cítrica está entre os produtos que podem ser utilizados como substitutos na alimentação de ruminantes, diminuindo os gastos e mantendo a qualidade nutricional do alimento fornecido aos animais, porém, esses alimentos devem ser utilizados de forma que não tragam malefícios. Assim, o presente relato visa apresentar a ocorrência de erosão dentária, actinomicose e polioencefalomalácia em ovinos criados e mantidos recebendo alimentação à base de polpa cítrica úmida despectinada na concentração de 50% do volumoso. A actinomicose foi diagnosticada em cinco animais por meio de exame clínico, radiográfico e cultivo microbiológico, e após tratamento três animais foram curados. Já a polioencefalomalácia foi confirmada em dez animais pelos sintomas manifestados, eficiência da terapia instituída em nove animais e exame anatomopatológico de um animal que veio a óbito. De acordo com o observado, deve-se ter cuidado ao utilizar grande quantidade de polpa cítrica úmida como volumoso.

Occurrence of acid erosion of the teeth, actinomycosis and polioencephalomalacia in flock of sheep fed wet low pectin citrus pulp / Ocorrência de erosão ácida dos dentes, actinomicose e polioencefalomalacia em rebanho de ovinos alimentados com polpa cítrica úmida despectinada

The citrus pulp can be used as a substitute in ruminant feed reducing costs and maintaining the nutritional quality of food. However, this compound should be used carefully so as not to cause harm to the animals. The present report aims to describe the occurrence of dental erosion, actinomycosis and polioencephalomalacia in sheep raised and kept with a wet low pectin citrus pulp based diet, composing 50% of roughage. Actinomycosis was diagnosed in five animals through clinical and radiographic examinations and microbiological culture, and, after treatment, three animals were cured. Polioencephalomalacia was confirmed in ten animals by clinical diagnostics, in nine out of ten animals by therapeutic diagnosis, and in one animal by post-mortem anatomopathological examination. According to the observed, we recommend caution when large amounts of citrus pulp are used as bulky food.(AU)

A polpa cítrica está entre os produtos que podem ser utilizados como substitutos na alimentação de ruminantes, diminuindo os gastos e mantendo a qualidade nutricional do alimento fornecido aos animais, porém, esses alimentos devem ser utilizados de forma que não tragam malefícios. Assim, o presente relato visa apresentar a ocorrência de erosão dentária, actinomicose e polioencefalomalácia em ovinos criados e mantidos recebendo alimentação à base de polpa cítrica úmida despectinada na concentração de 50% do volumoso. A actinomicose foi diagnosticada em cinco animais por meio de exame clínico, radiográfico e cultivo microbiológico, e após tratamento três animais foram curados. Já a polioencefalomalácia foi confirmada em dez animais pelos sintomas manifestados, eficiência da terapia instituída em nove animais e exame anatomopatológico de um animal que veio a óbito. De acordo com o observado, deve-se ter cuidado ao utilizar grande quantidade de polpa cítrica úmida como volumoso.(AU)

Occurrence of acid erosion of the teeth, actinomycosis and polioencephalomalacia in flock of sheep fed wet low pectin citrus pulp / Ocorrência de erosão ácida dos dentes, actinomicose e polioencefalomalacia em rebanho de ovinos alimentados com polpa cítrica úmida despectinada

The citrus pulp can be used as a substitute in ruminant feed reducing costs and maintaining the nutritional quality of food. However, this compound should be used carefully so as not to cause harm to the animals. The present report aims to describe the occurrence of dental erosion, actinomycosis and polioencephalomalacia in sheep raised and kept with a wet low pectin citrus pulp based diet, composing 50% of roughage. Actinomycosis was diagnosed in five animals through clinical and radiographic examinations and microbiological culture, and, after treatment, three animals were cured. Polioencephalomalacia was confirmed in ten animals by clinical diagnostics, in nine out of ten animals by therapeutic diagnosis, and in one animal by post-mortem anatomopathological examination. According to the observed, we recommend caution when large amounts of citrus pulp are used as bulky food.(AU)

A polpa cítrica está entre os produtos que podem ser utilizados como substitutos na alimentação de ruminantes, diminuindo os gastos e mantendo a qualidade nutricional do alimento fornecido aos animais, porém, esses alimentos devem ser utilizados de forma que não tragam malefícios. Assim, o presente relato visa apresentar a ocorrência de erosão dentária, actinomicose e polioencefalomalácia em ovinos criados e mantidos recebendo alimentação à base de polpa cítrica úmida despectinada na concentração de 50% do volumoso. A actinomicose foi diagnosticada em cinco animais por meio de exame clínico, radiográfico e cultivo microbiológico, e após tratamento três animais foram curados. Já a polioencefalomalácia foi confirmada em dez animais pelos sintomas manifestados, eficiência da terapia instituída em nove animais e exame anatomopatológico de um animal que veio a óbito. De acordo com o observado, deve-se ter cuidado ao utilizar grande quantidade de polpa cítrica úmida como volumoso.(AU)

Indicadores prognósticos para mastocitomas: estudo morfométrico e imunoistoquímico / Prognostic indicators for mast cell tumors: a morphometric and immunohistochemical study

Noventa e três mastocitomas cutâneos caninos, presentes em 74 animais, tiveram seus históricos clínicos investigados. Destes, 25 casos foram submetidos à análise morfométrica nuclear em lâminas de citopatologia nas colorações Panótico rápido e H&E; e 28 casos à graduação histopatológica e à imunoistoquímica para detecção das expressões de KIT, BAX, Ki-67 e PCNA, de modo a identificar os melhores indicadores prognósticos para a neoplasia. Os parâmetros citomorfométricos nucleares investigados foram: área, diâmetro-médio e perímetro. Os tumores foram classificados em graus histopatológicos I (bem diferenciados), II (de diferenciação intermediária) ou III (pobremente diferenciados), segundo a proposta de Patnaik, Ehler e MacEwen (1984). Os resultados obtidos através dos indicadores investigados foram comparados às graduações oferecidas pelo exame histopatológico, bem como aos tempos de sobrevida pós-cirúrgica e à mortalidade em função da doença. A citomorfometria obteve correlação com a mortalidade e o tempo de sobrevida dos animais em ambas as colorações testadas. Entre os marcadores imunoistoquímicos, as expressões de Ki-67 e BAX mostraram-se eficientes na previsão da sobrevida e mortalidade para o tumor, enquanto que as de KIT e PCNA não foram indicadores prognósticos para a neoplasia. Quanto maiores os parâmetros citomorfométricos e as porcentagens de núcleos positivos para Ki-67 e BAX, maiores as taxas de mortalidade e menores os períodos de sobrevida pós-cirúrgica. Finalmente, a graduação histopatológica foi o método que obteve os resultados mais expressivos na histopatologia, demonstrando ainda ser o método mais confiável deste painel. Nossos resultados apresentam um novo método de classificação para os mastocitomas cutâneos caninos, a partir da análise morfométrica aliada ao exame citopatológico, e confirmam que as avaliações imunoistoquímicas de Ki-67 e BAX, podem auxiliar na graduação dos mastocitomas, contribuindo com o estabelecimento de prognósticos e tratamentos mais precisos

Ninety-three canine cutaneous mast cell tumors, of 74 dogs, had their clinical history investigated. Among these, 25 cases were submitted to nuclear morphometric analysis on cytopathology slides stained by panoptic method and hematoxylin & eosin; and 28 cases were submitted to histopathologic grading and immunohistochemistry to evaluate KIT, BAX, Ki-67 e PCNA expressions, in order to identify the best prognostic indicators for this neoplasm. The nuclear cytomorphometric parameters used in the present study were: area, average-diameter and perimeter. The tumors were histologically classified as grades I (well differentiated), II (of intermediate differentiation) or III (poorly differentiated), as proposed by Patnaik, Ehler and MacEwen (1984). The results obtained by the indicators investigated were compared to the histopathologic grading, as well as to post-surgical survival times and mortality due to mast cell disease. Cytomorphometry was correlated with mortality and survival times in both staining methods. Among the immunohistochemical markers, Ki-67 and BAX expressions were efficient in predicting survival times and mortality due to mast cell tumors. KIT and PCNA expressions were not prognostic indicators for this tumor. The increases observed in the cytomorphometric parameters, and Ki-67 and BAX positive nuclei percentages were correlated with higher mortality rates and reduced post-surgical survival times. Finally, using the histopathologic grading system, we obtained the best results on histopathology, being this the most trustworhty method of this diagnostic panel. Our results presents a new grading system for canine cutaneous mast cell tumors, based on morphometric analysis on cytopathology slides, and confirm that immunohistochemical evaluation of Ki-67 and BAX expression may contribute to mast cell grading, leading to more accurate prognosis and treatment