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.

Accuracy of four ultrasonography techniques in predicting histopathological classification of canine mammary carcinomas.

Due to the importance of presurgical, noninvasive, and accurate diagnostic tools in mammary carcinoma characterization, this prospective secondary observational cohort study was designed to evaluate and compare the diagnostic accuracy of B-mode, Doppler, contrast enhancement ultrasonography, or acoustic radiation force impulse-elastography in identifying mammary carcinomas types with high degree of malignancy. A total of 246 mammary carcinomas from 141 female dogs were analyzed using B-mode, Doppler, contrast enhancement ultrasonography, and acoustic radiation force impulse ultrasonography prior to their histopathological classification according to types (simple, complex, or special) and grade (I, II, or III). Qualitative and quantitative variables were compared between carcinoma types and grades by Fisher's or analysis of variance. Diagnostic performance was estimated by receiver-operating characteristic analysis, using histopathological classification as a reference. Deformability (acoustic radiation force impulse) had a diagnostic specificity of 100% and sensitivity of 12% in identifying special carcinomas. A width:length ratio greater than 0.53 can be suggestive of special carcinoma, with 80% sensitivity and 76% specificity. Contrast wash-in and peak enhancement times lower than 7.5 and 13.5 s, respectively, were indicative of complex carcinoma at 62% sensitivity and 60% specificity. Contrast wash-in, peak enhancement, and wash-out times greater than 6.5, 12.5, and 64.5 s, respectively; were indicative of grade II and III carcinoma at 68% sensitivity and 62% specificity. In conclusion, B-mode ultrasonography, contrast enhancement ultrasonography, and acoustic radiation force impulse-elastography enabled the identification of some of the characteristics of high-grade mammary carcinoma types and grades in female dogs with limited accuracy. The findings from this study may contribute to oncology research and clinical management canine patients.

Efeitos de diferentes doses de hiclato de doxiciclina nos parâmetros hematológicos de cães com erliquiose / Effects of different doses of doxycycline hyclate on haematological parameters of dogs with ehrlichiosis

Este estudo objetivou comparar os efeitos de duas doses de hiclato de doxiciclina sobre a contagem de hemácias, concentração de hemoglobina, hematócrito, contagem de leucócitos e plaquetas de cães com erliquiose. Foram estudados os grupos I, composto por cães saudáveis (n=6), negativos na sorologia para Ehrlichia canis e Leptospira spp., na PCR em tempo real para E. canis e Anaplasma platys e na semi–nested PCR para Babesia canis; grupos II (n=6) e III (n=6), compostos por cães com história clínica sugestiva, sorologia e/ou PCR em tempo real positivas para E. canis, e negativos na pesquisa de anticorpos anti-Leptospira spp., na PCR em tempo real para A. platys e na semi–nested PCR para B. canis. Os cães doentes foram tratados com hiclato de doxiciclina via oral a cada 12 horas por 30 dias (5 mg/kg, grupo II; 10 mg/kg, grupo III). Hemogramas foram feitos antes, decorridos 15 dias e aos 10 dias após o término dos tratamentos. Não houve diferenças entre os grupos nos tempos avaliados quanto às variáveis hemácias, hemoglobina, hematócrito e leucócitos. Observou-se diferença quanto aos trombócitos entre o grupo I e os grupos II e III (p<0,0001) no início do estudo. Aos 15 dias de tratamento, a média do grupo III foi menor que dos grupos I (p=0,008) e II (p=0,0007), indicando persistência da trombocitopenia, ausente no grupo II. Não houve diferença entre os grupos no tempo final, sugerindo que ambos os tratamentos promoveram aumento da contagem de plaquetas em cães naturalmente infectados por E. canis.

This study aimed to compare the effects of two doses of doxycycline hyclate on red blood cells, hemoglobin, hematocrit, white blood cells and platelets of dogs with ehrlichiosis. Group I, comprised of healthy dogs (n=6), negative on serology for Ehrlichia canis and Leptospira spp., real time PCR forE. canis and Anaplasma platys, and on semi–nested PCR for Babesia canis; Groups II (n=6) and III(n=6), comprised of dogs with suggestive clinical history, positive serology and/or real time PCR for E.canis, negative on research for anti-Leptospira spp. antibodies and real time PCR for A. platys, and on semi–nested PCR for B. canis were studied. Sick dogs were treated with doxycycline hyclate every 12hours, by mouth, for 30 days (5 mg/kg, group II; 10 mg/kg, group III). Complete blood counts were performed before, after 15 days, and 10 days after period of treatment was complete. No difference between groups at the studied time points were noticed for red blood cells, hemoglobin, haematocrit and white blood cells. Difference was observed for platelets between group I and groups II and III(p<0.0001) at the study on set. After 15 days of treatment, the mean platelet for group III was lower than groups I (p=0.008) and II (p=0.0007), indicative of persistent thrombocytopenia, already absentin group II. No difference between groups was noticed at final time point, which suggests that both treatments increased platelets in dogs naturally infected with E. canis.

Avaliação do sulfato de dehidroepiandrosterona (DHEA-S) sérico em cães acometidos por hipercortisolismo espontâneo / Evaluation of serum dehydroepiandrosterone sulfate (DHEA-S) in dogs affected by spontaneous hypercortisolism

O hiperadrenocorticismo (HAC) ou Síndrome de Cushing é uma endocrinopatia comum em cães e caracteriza-se por aumento na secreção de cortisol e ou de outros hormônios também secretados pela cortical da adrenal. Existem diferentes testes diagnósticos que visam avaliar as concentrações séricas de cortisol, mas em alguns casos os resultados mostram-se duvidosos ou negativos mesmo em animais com clínica evidente da doença. O tratamento de escolha para o HAC ACTH dependente é com o trilostano, medicamento que inibe a produção do cortisol. O sulfato de dehidroepiandrosterona (DHEA-S) é um precursor esteroide sintetizado a partir do colesterol e a avaliação de sua concentração em cães com HAC recebendo ou não o tratamento com o trilostano pode trazer importantes informações sobre seu comportamento nessa doença. Sendo assim, objetivou-se neste estudo avaliar as concentrações de DHEA-S em animais com HAC ACTH dependente sem tratamento e tratados com trilostano, a fim de elucidar seu comportamento nessas diferentes situações, buscando um meio alternativo de diagnóstico para o HAC. Vinte e cinco cães foram separados em 3 grupos, de modo que o grupo A (n=6) foi composto por cães saudáveis, sem evidência de qualquer doença sistêmica; o grupo B (n=10) foi composto por cães diagnosticados com HAC ACTH dependente; e animais em tratamento com trilostano compuseram o grupo C (n=9). Todos os cães foram submetidos ao teste de estimulação com ACTH para dosagem de cortisol e DHEA-S. Os resultaram mostraram que animais em tratamento com trilostano apresentam médias de DHEA-S maiores do que os cães saudáveis e os com HAC sem tratamento. O que evidencia a inibição enzimática realizada pelo trilostano na cascata de produção do cortisol e dos hormônios sexuais. Não...

The hyperadrenocorticism (HAC) or Cushing's syndrome is a common endocrine disease in dogs and is characterized by an increase in cortisol secretion and or other hormones also secreted by the adrenal cortex. There are variety of diagnostic tests aimed at evaluating the serum concentrations of cortisol, but in some cases the results are unclear or negative even in animals with evident clinical disease. Trilostane is the gold standard treatment for HAC dependent-ACTH, that drug inhibits the production of cortisol. Dehydroepiandrosterone sulfate DHEA (S) is a steroid precursor synthesized from cholesterol and evaluation of their concentration in dogs with HAC receiving or not treatment with trilostane may provide important information about their behavior in this disease. Thus, the aim of this study was to evaluate the concentrations of DHEA-S in animal with HAC ACTH dependent untreated and treated with trilostane, in order to elucidate their behavior in these different situations, seeking an alternative means of diagnosis for HAC. Twenty-five dogs were divided into 3 groups. The Group A (n = 6) was composed of healthy dogs with no evidence of any systemic disease; group B (n = 10) was composed of dogs diagnosed with HAC ACTH-dependent; and animals treated with trilostane comprised group C (n = 9). All dogs were submitted to ACTH stimulation test. The results showed that animals treated with trilostane have higher DHEA-S medium than healthy dogs with HAC and without treatment. This result showed the enzyme inhibition from the trilostane in cortisol production cascade and sex hormones. No significant differences were observed in mean DHEA-S among the animals of groups A and B. This shows that animals with untreated HAC not have higher DHEA-S values than healthy animals, as was expected. In addition, the measurement of DHEA-S was not adequate for diagnosis of dogs with HAC.