Expression of acetylated histones H3 and H4 and histone deacetylase enzymes HDAC1, HDAC2 and HDAC6 in simple mammary carcinomas of female dogs.

Histone deacetylation is an important mechanism involved in human breast cancer tumorigenesis and recent veterinary oncology studies also demonstrate a similar relationship in some canine neoplasms. The use of HDAC inhibitors in vitro and in vivo has demonstrated antitumor action on several strains of human and animal cancers. The present study aims to correlate the expression of H3K9Ac, H4K12Ac, HDAC1, HDAC2 and HDAC6 in simple mammary carcinomas in dogs with clinicopathological parameters and overall survival time. To this end, 61 samples of simple breast carcinomas were analyzed by the immunohistochemistry technique with subsequent validation of the antibodies by the Western Blot technique. The expressions obtained via a semi-quantitative way were categorized by assigning scores and classified into high or low expressions according to the given score, except for HDAC6, when the marking percentage was considered and subdivided into high and low expressions using the median value. For statistical analysis, the chi-square test or Fisher exact test were used as univariate analysis and correspondence analysis as a multivariate test, in addition to the Kaplan-Meier survival analysis. In the studied samples, the highest frequencies were determined for the high expression proteins H4K12Ac (88.5%), HDAC2 (65.6%) and HDAC6 (56.7%) and the low expression proteins H3K9Ac (73.8%) and HDAC1 (54.1%). An association between the low expression of HDAC1 and the presence of lymph node metastasis (p = 0.035) was indicated by univariate analysis while the high expression of HDAC1 was associated with favorable prognostic factors, such as the absence of lymph node metastasis and low mitotic index by multivariate analysis. Also, by multivariate analysis, the low expression of HDAC6 was correlated with the low expression of Ki67, smaller tumors, and better prognosis factors as well. Protein expression was not correlated with patients' overall survival time (p > 0.05). The high expressions of HDAC2 and HDAC6 in mammary carcinomas in female dogs may be useful information for research involving therapeutic targets with iHDACs since their inhibition favors hyperacetylation and transcription of tumor suppressor genes.

Toxicity of a methotrexate metronomic schedule in Wistar rats.

Metronomic chemotherapy is a relevant strategy that uses low doses of antineoplastic drugs for sustained periods to control tumor growth, an alternative frequently utilized in veterinary patients. This work aimed to evaluate the toxic effects of a metronomic oral dose of methotrexate (MTX) for 45 days in tumor-free Wistar rats when compared with control animals. Clinical alterations, body weight, food, and water intake were monitored daily, and bone marrow suppression, hematological, biochemical, and histopathological analyses were performed at three points (days 30, 45, and 60). MTX-treated animals did not demonstrate severe systemic involvement. At 30 days, compared with control animals, MTX-treated animals showed significant leukocytosis (11.9 ± 2.3 vs. 7.8 ± 0.2 106/µL; P < .05) and augmentation of immature myeloid populations from bone marrow (9.0 ± 0.8 vs. 6.5 ± 1.5%; P < .05), and at 60 days, treated animals showed significant neutrophilia (35.0 ± 11.0 vs. 23.00 ± 3.0%; P < .05), depletion of bone marrow lymphocytes (8.2 ± 0.7 vs. 11.5 ± 1.9%; P < .05), and immature myeloid populations (7.2 ± 0.7 vs. 8.3 ± 0.6%; P < .05). At a histopathological level, splenic hypoplasia and respiratory inflammatory lesions were significant when compared with control animals, presenting mild to moderate myelotoxicity, immune suppression, and associated clinical compromise that persisted beyond treatment withdrawal. This suggested that MTX metronomic toxicity should not be neglected owing to the observed residual side-effects and special care should be taken regarding myelosuppression.

Proteinograma sérico e do líquido peritoneal de equinos hígidos e daqueles submetidos à obstrução intestinal experimental / Serum and peritoneal protein profile of healthy horses and horses subjected to experimental intestinal obstruction

Avaliou-se a resposta de fase aguda através da concentração das proteínas de fase aguda (PFA) no soro sanguíneo e no líquido peritoneal de vinte e um equinos, hígidos e submetidos à obstrução intestinal experimental, distribuídos em quatro grupos: obstrução de duodeno - GD (n=6), íleo - GI (n=6), cólon dorsal esquerdo - GM (n=6) e controle instrumentado - GC (n=3). Foram colhidas amostras de sangue e líquido peritoneal e, após centrifugação e fracionamento, as proteínas de fase aguda foram separadas por eletroforese em SDS-PAGE. Identificaram-se as proteínas IgA, ceruloplasmina, transferrina, albumina, IgG, haptoglobina, α1-glicoproteína ácida e P24, no soro e no líquido peritoneal. Houve aumento nas concentrações sérica e peritoneal de todas as PFA, sendo mais evidente no líquido peritoneal e nos animais obstruídos. O fracionamento eletroforético das PFA no líquido peritoneal é mais eficaz no diagnóstico de processos inflamatórios abdominais, quando comparado ao sérico.

It was evaluated the acute phase response over the concentration of acute phase proteins (APP) in serum and in peritoneal fluid of twenty-one healthy equines submitted to experimental intestinal obstruction distributed in groups: duodenal obstruction (GD), ileum obstruction (GI), left-dorsal colon (GM), and instrumentalized control (GC). Blood and peritoneal liquid samples were collected, after centrifugation and fractioning and the acute phase proteins were separated by electrophoresis in SDS-PAGE. The acute phase proteins (APP) identified were IgA, ceruloplasmin, transferrin, albumin, IgG, haptoglobin, α1-acid glycoprotein and P24, in the serum and liquid peritoneal. There was anincrease in the serum and peritoneal proteins concentrations evidencing a higher response of the APP from the peritoneal fluid and of the obstructed groups. The electrophoretic separation of the proteins included in the peritoneal liquid is more effective when compared to the serum concentration in the diagnosis of abdominal inflammatory processes.

Anomalia de Pelger Huet em animais domésticos: uma revisão / Pelger Huet Anomaly in pet animals: a review

A anomalia de Pelger Huet (APH) é uma doença hereditária caracterizada pela não segmentação nuclear dos granulócitos, especialmente dos neutrófilos, devido a defeito genético no receptor da lamina-B (RLB) no cromossomo 1q41-43. Os neutrófilos apresentam núcleos bilobulados, ovais ou arredondados e citoplasma abundante, caracterizando assim um pseudo desvio à esquerda degenerativo. Neste trabalho revisional, objetivou-se caracterizar e descrever a anomalia em animais domésticos. A anomalia já foi descrita em cães, gatos, coelhos e equinos. Em todos os casos a APH mostrou-se como um achado hematológico ocasional, pois os pacientes foram levados à clínica veterinária devido a outras enfermidades e, após antibioticoterapia, descarte de doenças infecciosas e ausência de sinais clínicos; a persistência da hipossegmentação dos neutrófilos, durante meses, levou à conclusão de que se tratava de casos de APH. A hipossegmentação, nesses casos, foi confirmada pelo grau de segmentação do núcleo dos granulócitos. Apesar da falta de segmentação do núcleo, estudos demonstraram que não há alteração na função dos neutrófilos de cães com essa anomalia. A análise comparativa dos granulócitos de cães normais e com APH, através da microscopia eletrônica, coloração citoquímica e citometria de fluxo não evidenciaram diferenças significativas entre os dois grupos. Assim, a APH é considerada um achado ocasional e benigno do exame de sangue, cujas células devem ser diferenciadas morfologicamente de formas imaturas de neutrófilos presentes, principalmente, nos casos de enfermidades como doenças inflamatórias e leucemias.

Pelger Huet anomaly (PHA) is a hereditary disease characterized by non-segmentation of the granulocyte nuclei, especially neutrophils, due to a genetic defect in the lamin B-receptor (LBR) in chromosome 1q41-43. In that disorder, neutrophils present bilobulated, oval or round nuclei, and abundant cytoplasm, which characterizes a pseudo-degenerative left shift. The aim of this review was to characterize and describe PHA in pet animals. The anomaly has been described in dogs, cats, rabbits and horses, and all cases, it has shown itself as an occasional hematological finding, once animals were seen by the veterinarian because of other disorders. The persistence of neutrophils hyposegmentation for months after antibiotic treatment, ruling out infectious diseases, and absence of clinical signs led to PHA diagnosis. Hyposegmentation in those cases was confirmed by the degree of segmentation in granulocyte nuclei. However, despite the absence of nucleus segmentation, studies have shown that neutrophil function in PHA-affected dogs remains normal. The use of electron microscopy, cytochemical staining and flow cytometry did not show any significant differences between animals affected by PHA and animals considered normal. PHA is considered an incident and benign finding in blood test, whose cells should be morphologically differentiated from immature neutrophils, especially in cases of inflammatory diseases and leukemia.

Correlação da atividade de lactato desidrogenase e concentração de lactato com a classificação de efusões em cães / Correlation of lactate dehydrogenase and lactate concentration with dog's effusion classification

As efusões são problemas clínicos frequentes e que ocorrem em consequência de uma enfermidade que culmine com diminuição da pressão coloidosmótica intravascular, elevação da pressão hidrostática local, aumento da permeabilidade vascular e/ou comprometimento da drenagem realizada pelos vasos linfáticos. Dessa maneira, a avaliação laboratorial desse fluido torna-se relevante para que, em conjunto com os sinais clínicos apresentados pelo paciente, possa ser firmado um possível diagnóstico e instituída ação terapêutica adequada. Assim sendo, a classificação de uma efusão em transudato ou exsudato torna-se um dos pontos críticos para a elucidação do diagnóstico e condução do caso clínico. Em Medicina Veterinária, o método tradicional de classificação de uma efusão é baseado na contagem celular e na concentração de proteínas do fluido. Contudo, diversos estudos evidenciam que tais parâmetros não são suficientes para a correta classificação de todas as efusões. Assim, o presente estudo foi conduzido com o objetivo de verificar a correlação de outros parâmetros bioquímicos com a diferenciação das efusões transudativas e exsudativas e, para tal, foram avaliadas as atividades de lacatato desidrogenase (LDH) das efusões, a relação de sua atividade fluido/soro, concentração de lactato das efusões, o gradiente de concentração de lactato do soro para o fluido e a concentração de proteínas das efusões. Os resultados obtidos permitiram observar que a atividade de LDH, a relação LDH efusão/soro, a concentração de lactato e o gradiente de concentração de lactato soro/efusão apresentam diferença estatisticamente significativa (P<0,05), bem como alta correlação com a classificação de uma efusão exsudativa.

The effusions are frequent clinical problems and can occur in consequence to an illness that culminates with decrease of the intravascular coloidosmotic pressure, increase of local hydrostatic pressure, increase of vascular permeability and/or compromising of the drainage accomplished by lymphatic vases. Therefore, the laboratorial evaluation of this fluid becomes relevant, jointly with clinical signs presented by patient; to become possible the diagnosis definition and institution of appropriate therapeutic. Thus, classification of effusion in exudate and transudate is one of major points to elucidation of diagnosis and conduction of clinical case. In veterinary medicine the traditional method of an effusion classification is based on cellular counting and protein concentration of the fluid, however, several studies evidence that such parameters are not enough for the correct classification of all kinds of effusions. Considering this, the present study aimed to verify the correlation of some biochemical parameters with the differentiation of transudatives and exudatives effusions. To perform this, the activities of lactate dehydrogenase (LDH) were appraised, as well as the relationship of their activities with fluid/serum; lactate and proteins concentration and fluid/serum gradients of concentration of these same substances. The results allowed to observe that the activity of LDH, relationship LDH and fluid/serum, lactate concentration and lactate fluid/serum gradient of concentration present statistically significant difference (P<0.05), as well as a high correlation with the classification of an exudative effusion.

Aspectos clínicos e concentração sérica da creatina-quinase e lactato-desidrogenase em cães submetidos à fisioterapia após atrofia muscular induzida / Clinical aspects and serum concentration creatina kinase and lactate dehydrogenase in dogs submitted to physiotherapy after induced muscle atrophy

Avaliou-se a resposta de diferentes protocolos fisioterapêuticos em cães após a indução de atrofia muscular por meio da imobilização do joelho por 30 dias. Os grupos foram denominados grupo C ou controle, grupo E (massagem, movimentação passiva e eletroterapia), grupo H (massagem, movimentação passiva e hidroterapia em esteira aquática) e grupo EH (massagem, movimentação passiva, eletroterapia e hidroterapia em esteira aquática). Foram mensurados os graus de claudicação, arco do movimento, circunferência da coxa e a variação sérica das enzimas creatina-quinase e lactato-desidrogenase. De acordo com os resultados encontrados, foi possível concluir que as modalidades terapêuticas de massagem, movimentação passiva da articulação, estimulação elétrica neuromuscular e hidroterapia por caminhada em esteira aquática aceleram a recuperação clínica em cães com atrofia muscular induzida.

The response of different physiotherapeutic treatment protocols was evaluated in dogs after muscle atrophy induced by joint immobilization for 30 days. Groups were named C group or control, E group (massage, passive range of motion and neuromuscular electrical stimulation), H group (massage, passive range of motion and aquatic therapy in underwater treadmill) and EH group (massage, passive range of motion, neuromuscular electrical stimulation and aquatic therapy in underwater treadmill). It was measured the degree of lameness, range motion, thigh circumference and range of serum creatine kinase (CK) and lactate dehydrogenase (LDH). According to the results, it was possible to conclude that associated therapeutics modalities such as massage, passive range of motion of the joint, neuromuscular electrical stimulation and aquatic therapy by walking on underwater treadmill accelerate clinical recovery in dogs with induced muscle atrophy.