EVALUATION OF CYTOTOXICITY AND EXPRESSION OF CASPASE-3 AND P53 IN HCT-116 CELLS OF LINEAGE TREATED WITH DIFFERENT EXTRACTS OF Euphorbia tirucalli L.

Euphorbia tirucalli, a plant commonly known as aveloz, has been employed in anticancer therapy with little scientific proof. The goal of this study was to evaluate the cytotoxic potential and expression of caspase-3 and p53 in colorectal cancer cell lines treated with aveloz extracts. Hexanic and hydroalcoholic extracts were prepared in different concentrations. After 96 h, cell viability and apoptosis rate (p53 and caspase-3 expression) were verified. The hexanic extract revealed higher cytotoxicity on HCT-116 cell line at concentrations above 125 µg/mL, and hydroalcoholic extract above 500 µg/mL. LC50 for the hexanic extract was 60.2873 µg/mL, and for hydroalcoholic extract 100.6066 µg/mL. There were significant differences among the averages of cells marked with p53 at the concentrations 62.5, 250 and 500 µg/mL, as well as among the averages of caspase-3-positive cells at the concentrations 62.5, 125 and 500 µg/mL, with higher cytotoxic effect being observed for the hydroalcoholic extract. The obtained data provides evidence for the cytotoxic potential in vitro of the hydroalcoholic and hexanic extracts of aveloz on HCT-116 cell line. There was an increase in the expression of caspase-3 and p53 in the tumor line cells exposed to extracts of E. tirucalli

HISTOPATHOLOGY ASPECTS AND FIBROSIS EVALUATION OF BOVINE NATURALLY INFECTED LIVERS BY Fasciola hepatica

Fasciolose é uma zoonose que infecta especialmente os ruminantes e os danos ao fígado causado pela infecção crônica pelo parasito são graves. Objetiva-se com este estudo avaliar os achados histopatológicos no fígado de bovinos naturalmente infectados com Fasciola hepatica, bem como propor a classificação de lesão hepática causada por essa doença baseado na inflamação crônica e fibrose. Foram utilizados fragmentos do lobo direito e esquerdo de 100 fígados bovinos com fasciolose. Os fragmentos foram submetidos a processamento histológico e coloração com HE e Tricrômico de Masson. A classificação das lesões foi realizada por meio da atribuição de uma classificação gradual de I a III, de acordo com a intensidade das alterações inflamatórias e fibróticas. As lesões mais prevalentes foram a proliferação do ducto biliar, infiltração de células mononucleares, arteriosclerose e fibrose periportal e periductal. Lesões fibróticas foram mais intensas do que a inflamação e 45% das amostras pertenciam à categoria II, seguido de 39% na categoria I e 16% na categoria III. A proposta de categorização histopatológica da lesão hepática em fasciolose crônica em bovinos naturalmente infectados provou ser um método valioso de avaliação microscópica, uma vez que permite determinar a evolução da doença

CARACTERIZAÇÃO HISTOLÓGICA DE FÍGADOS BOVINOS COM FASCIOLOSE CRÔNICA

A fasciolose bovina produz danos graves ao fígado e é causada pelo parasito Fasciola hepatica. Objetivou-se caracterizar as alterações histológicas do tecido hepático em bovinos com fasciolose crônica. Foram coletados 47 fígados de bovinos condenados por fasciolose e 12 fígados normais utilizados para controle. Foi feita a contagem de parasitos e coleta de cinco fragmentos de cada lobo, direito e esquerdo, dos fígados acometidos pela enfermidade, totalizando 235 amostras por lobo. O grupo controle foi coletado apenas um fragmento por fígado. Todo material foi fixado em formol a 10% e submetido ao processamento histológico. Foi feita a análise microscópica com base na distribuição da fibrose em graus 1, 2 e 3 e no tipo e intensidade do infiltrado inflamatório. A avaliação estatística foi feita pelo método descritivo e pelo teste de Mann-Whitney com 5% de probabilidade. Não foi verificada diferença entre o número de parasitos por lobo hepático, porém o lobo esquerdo apresentou maior intensidade parasitária. A fibrose foi observada em todas as amostras prevalecendo a de grau 1. Houve diferença significativa entre fígados com F. hepatica e fígados normais quanto ao grau de fibrose, tanto para o lobo direito quanto para o esquerdo e entre estes. O infiltrado inflamatório revelou predominância de células mononucleadas de intensidade discreta. Houve diferença entre a intensidade do infiltrado inflamatório entre os lobos direito e esquerdo do fígado. A presença de eosinófilos também foi verificada, porém em menor quantidade. Conclui-se que a fibrose é alteração mais marcante em fígados cronicamente infectados por F. hepatica e é mais evidente no lobo hepático esquerdo. O infiltrado inflamatório é predominantemente mononuclear discreto e também mais prevalente no lobo esquerdo

IDENTIFICAÇÃO DE PARASITOS E ACHADOS MACROSCÓPICOS DE NECROPSIA EM PINGUINS-DE-MAGALHÃES NO ESPÍRITO SANTO, BRASIL

Pinguins são aves marinhas extremamente bem adaptadas à vida no mar, mas não é raro encontrar indivíduos mortos ao longo da costa e as mortes são causadas por fatores relacionados tanto à interferências humanas no meio ambiente quanto por doenças específicas. Objetivou-se com este estudo identificar espécies de parasitos encontradas em pinguins-de-Magalhães na costa do Espírito Santo, Brasil, e descrever os principais achados macroscópicos de necropsia. Cinco aves apresentaram ectoparasitos. Todas as aves tinham nematodeos. Quatro indivíduos tinham trematodeos. Todos os pinguins estavam com escore de condição corporal 2. Dois dos indivíduos examinados apresentavam ruptura de esôfago por corpo estranho e um indivíduo apresentou corpo estranho no estômago. No sistema respiratório, as lesões encontradas foram congestão pulmonar, fibrina nos sacos aéreos, conteúdo caseoso na traqueia, edema pulmonar e pneumonia. Pinguins-de-Magalhães na costa do estado do Espírito Santo são suscetíveis a parasitoses. Helmintos das espécies Contracaecum pelagicum e Cardiocephaloides physalis foram identificados, bem como ectoparasitos da espécie Austrogoniodes bifasciatus. As principais lesões macroscópicas encontradas estavam localizadas no sistema respiratório e no trato gastrointestinal

ZONA ORGANIZADORA DE NUCLÉOLO E SUA RELAÇÃO COM A EPIDEMIOLOGIA DE CADELAS COM NEOPLASIA MAMÁRIA

As neoplasias mamárias são comumente encontradas em cadelas, sendo observada, geralmente, em animais senis, não castrados e sem predisposição racial. A técnica de AgNOR pode auxiliar no diagnóstico e prognóstico de tumores mamários. Objetivou-se relacionar o número de AgNOR aos dados epidemiológicos como idade de acometimento, raça do animal, status reprodutivo, status vivo ou morto, diagnóstico, recidiva e metástase. Fez-se um levantamento de dados de 49 cadelas com neoplasia mamária durante o período de 2011 a 2015 da rotina do HOVET-UFES e a técnica de AgNOR foi empregada em lâminas de tumores mamários de cadelas. Observou-se média de idade de 10 anos e contagem média de AgNOR correspondente a 4,42. As cadelas SRD foram as mais frequentes com média de AgNOR 4,22. O status vivo correspondeu a 63,64% das cadelas e a média de AgNOR foi 4,23, e as mortas, 36,36% com média de AgNOR de 4,75. Encontrou-se 90,90% de cadelas castradas, com média de AgNOR de 4,31 e não castradas 9,09%, com média de AgNOR 5,48. Neoplasias malignas foram observadas em 40 (81,63%) cadelas cuja média de AgNOR foi 4,46, e as benignas em nove (18,37%) com média de AgNOR de 4,24. Das cadelas, 42 (85,71%) não apresentaram recidiva tumoral, cuja média de AgNOR foi de 4,39, e em duas que apresentaram metástase tumoral, média de AgNOR de 5,04. Concluiu-se que o AgNOR varia em relação a idade e raça, e que também não houve dependência do status de sobrevida em relação ao número de AgNOR, bem como em relação ao status reprodutivo. Ainda, que o número de AgNOR variou conforme o diagnóstico e foi maior em tumores malignos e quando houve metástase

Morphometric and Vascular Analyses and MMP-2 Expression in Bladders of Animals with Bovine Enzootic Haematuria

Background: Bovine enzootic haematuria is a chronic, non-infectious disease caused by the ingestion of bracken Pteridium aquilinum that contain ptaquiloside, substance responsible for the development of neoplasms in combination with bovine papillomavirus infection. Studies examining metalloproteinases have shown that these enzymes may be useful as prognostic markers and many studies have correlated the intratumoural microvessel density and incidence of metastasis in a variety of cancers, including bladder cancer. This study aimed to evaluate the urothelial morphology and vasculature and quantify the expression of the enzyme metalloproteinase-2 in the bladder lesions of bovine with enzootic haematuria.Materials, Methods & Results: Forty bladders with macroscopic lesions were subjected to routine histological processing. The material was stained using the Hematoxylin-Eosin (HE) and Weigert's resorcin-fuchsin staining methods. The morphometric analysis included the measurement of the total thickness of the urothelium. Microscopically, the lesions were differentiated as neoplastic or non-neoplastic, and the vascular changes were characterised as vascular proliferation, ectasia, dilation and thickening and the material stained using the Weigert's resorcin-fuchsin method was used to evaluate sclerosis and the vascular fibroelastosis. The metalloproteinase expression was evaluated using an anti-metalloproteinase-2 antibody. The main non-neoplastic lesions found in the urothelium included clear cell metaplasia, dysplasia, hyperplasia, haemorrhage, cystitis cystica and Brunn's nests and the neoplastic lesions were haemangioma, myxoma, transitional cell carcinoma, adenocarcinoma, in situ carcinoma and haemangiosarcoma. The urothelium measurements revealed differences between the epithelial thicknesses of bladders with and without neoplasms. The bladder vasculature evaluation revealed the frequent occurrence of different pathological aspects, such as vascular proliferation, dilation, expansion, ectasia and thickening; more severe vascular proliferation, thickening and fibroelastosis were observed in bladders with neoplasms. Metalloproteinase-2 expression was observed in the epithelial cells, fibroblasts, endothelial cells and smooth muscle cells present in the tumour tissue and was a significant increase in the expression of MMP-2 in the neoplasms of mesenchymal origin compared to the neoplasms of epithelial origin.Discussion: The presence of a large variety of non-neoplastic lesions indicates the possibility that there are other factors concomitant to BEH involved. Epithelial hyperplasia and metaplasia, for example, could be the result of damage caused by bovine papillomavirus type 2 or even an altered urine pH. Furthermore, it is believed that the bladder has the capacity to generate various types of neoplasms due to the different histological components of the bladder, thus enabling it to generate both mesenchymal and epithelial lesions. The thickness of the urothelium was greater in all the quadrants with neoplasms compared to the quadrants without neoplasms. These data revealed that it is possible to use less invasive diagnostic imaging methods to characterise bovine enzootic haematuria and to measure tumour progression based on the severity of vascular changes. The vascular proliferation and thickening were significant in bladders with neoplasms, which may be explained by the fact that tumours require an extensive vascular supply. In addition, many tumours release vascular growth factors and promote neovascularisation. The intensity of the immunostaining of metalloproteinase-2 differed among the different types of neoplasms. However, due to the diversity of the tumours, it was not possible to establish a relationship between metalloproteinase-2 expression and tumour progression

Biometry of Adrenal Glands of Mixed-Breed Dogs

Background: Differences of the size of the adrenal glands, in healthy dogs or in endocrinopathies, undermine correct diagnosis of endocrine disorders and evaluations of the adrenal cortex in relation to its size and possible correlation with endocrinopathies are rare. The aim of the present study was to perform measurements of the length, width, thickness and weight of the adrenal glands of young, adult and elderly mixed-breed dogs and correlate them with the age, sex and weight of animals. In addition, the areas occupied by the zona glomerulosa, zona fasciculata, and zona reticularis of the adrenal cortex were measured in order to establish a microscopic biometric pattern.Material, Methods & Results: The right and left adrenal glands of 12 young (six females and six males), 12 adults (six females and six males), and 12 elderly (six females and six males), all mongrels, derived from routine and necropsied in the Animal Pathology Sector of the Veterinary Hospital of the Federal University of Espírito Santo (HOVET-UFES) were weighed and the length, width, and thickness were measured. For the microscopic measurement of the adrenal cortex, 10 randomly selected samples were submitted to routine histological processing and the microscope slides were observed under a light microscope at 5× and 10× objectives, photodocumented and measurements were obtained from a random portion of the adrenal cortex and the zones composing the cortex were measured in triplicate with the aid of the computerized image analysis software. The left adrenal showed a greater average length than the right adrenal in young, adult, and elderly dogs. The size of the glands in the young and adult dogs is not influenced by the sex of the animals, but in older dogs the females had a greater mean width than the males. The weight of the animals presented a positive correlation in relation to the length and weight of the right and left adrenal glands in all studied groups. The age did not influence the length and weight variables of the glands. However, some differences in thickness and width were observed in the elderly group compared to those in other groups. For the microscopic measurements in the right adrenal gland, the cortex was 1.53 mm, being 0.21 mm for the glomerular zone (14.6% of the total adrenal cortex), 1.04 mm for the zona fasciculata (66.9%), and 0.29 mm for the zona reticularis (18.5%). In the left adrenal gland, the cortex was 1.83 mm, being 0.23 mm for the glomerular zone (13.2% of the total adrenal cortex), 1.23 mm for the zona fasciculata (63.96%), and 0.37 for the zona reticularis (22.84%).Discussion: Studies on the size of the adrenal gland in dogs using macroscopic biometrics are scarce, and the current study presents results regarding mixed-breed (male and female) dogs of different ages, which presented variation in size and weight, which could also influence the size of the adrenal gland. Thus, it can be observed that a variation of measurements can be found, especially for dogs with no defined breed. These results demonstrate the importance of studies that perform the macroscopic analysis of such glands. It was concluded that there was a clear variation in the size of the adrenal glands in mixed-breed dogs of different ages and both sexes; the left adrenal showed greater lengths and weights than the right gland. The sex of the animals did not influence the size of the glands in young and adult dogs, but in elderly dogs, the females showed a greater mean width than the males. The right and left adrenal cortices are formed at a greater percentage by the zona fasciculata, followed by the zona reticularis and zona glomerulosa

Long bone fractures in Cerdocyon thous: macroscopic and microstructural evaluation

The aim of the present study was to perform the macroscopic and microstructural morphological classification of long bone fractures of Cerdocyon thous. Eighteen cadavers of the species were necropsied, and subjected to radiographic and microscopical evaluation when long bone fractures were detected. Among the 18 cadavers, eight (44%) had fractures equally distributed (33.33%) in the femur, humerus, or tibia. More frequently (61.54%), the fractures were simple and affected the diaphysis, and in smaller proportions (23.08%) reached the physeal line. In diaphyseal and metaphyseal fractures, microscopical evaluation revealed cortical bone tissue, with longitudinal osteons that contained longitudinal and intermediate collagen fibres and lamellae with a delamination aspect. On the other hand, in epiphyseal fractures, trabecular bone tissue was more frequently observed, consisting of trabeculae with disorganised collagen fibres and absence of osteons. In both cases low activity, osteocytes, and low coverage of osteoblasts on the bone surface were noted. It was concluded that the frequency of fractures in the long bones of C. thous was 44%, with females being more predisposed. The findings support the hypothesis that fractures in such animals are caused by being run over by automobiles. The present study contributes significantly in alerting clinicians and surgeons to the types of fractures that C. thous is more predisposed to, its places of greatest occurrence, and its microstructure. Thus, there is a need for joint actions aimed at reducing the number of cases of wild animals being run over by automobiles. Keywords: Bone biology. Orthopaedics. Run-over accidents. Trauma

Ultrassonografia quantitativa do fígado em gatos com tirotoxicose induzida

Este estudo avaliou ultrassonograficamente o fígado de gatos submetidos à tirotoxicose induzida pela técnica de histograma dos níveis de cinza com o objetivo de melhor avaliar possíveis alterações hepáticas decorrentes do excesso de hormônios tiroidianos. Para isto, foram utilizados nove gatos hígidos, adulto-jovens, que receberam diariamente 150µg/kg de levotiroxina sódica, por 60 dias. O histograma hepático não demonstrou diferença estatística ao nível de 5% de significância entre os momentos avaliados; entretanto, as variações dos valores das variáveis estudadas sugeriram a promoção de aumento da heterogeneidade e diminuição da ecogenicidade do parênquima hepático quando comparado ao momento inicial. A avaliação citopatológica e histopatológica revelou quadro de hepatite tirotóxica em todos os animais.This study evaluated ultrasonographically the liver of cats submitted to induced thyrotoxicosis by the technique of gray-level histogram with the objective of better evaluating the possible hepatic alterations decurring from the excess of the thyroid hormones. For that, there were used nine adult-young healthy cats, which received daily 150ìg/kg of sodic levotiroxin, for 60 days. The hepatic histogram has not demonstrated statistical differences in the 5% level of significance between the evaluated moments; however, the variations of the studied variables' values suggested an increasing of the heterogeneity and a decreasing of the echogenicity of the hepatic parenchyma when compared to the initial moment. The cytopathologic and histopathologic evaluations have shown of a clinical picture of thyrotoxic hepatitis in all the animals

Euterpe edulis pulp products and your effect on liver and kidney of in vivo model of colorectal cancer: Histopathological analysis / Produtos da polpa de Euterpe edulis e seus efeitos no fígado e nos rins em modelo in vivo de câncer colorretal: Análise histopatológica

The objective was to report the injuries on liver and kidney promoted by experimental colorectal carcinogenesis induction in rats and evaluate the effect of supplementation with Euterpe edulis M. pulp products on resolution of this injuries. Colorectal carcinogenesis with 1,2-dimethylhydrazine was induced in young male rats, allocated into: C - induced to carcinogenesis; CJ - induced to carcinogenesis and supplemented with juçara fruit pulp; and CE - induced to carcinogenesis and supplemented with juçara fruit lyophilized extract. Nine animals were a negative control. Supplementation occurred three times a week, totaling 54 days of administration with 1 mg of cyanidin-3-glycoside per kilogram live weight. The hepatic and renal histopathological injuries were assessed at 10 and 23 weeks. In liver, at 10-week biliary hyperplasia was more evident in colorectal cancer induced groups compared to N group (p = 0.0230), as well as megalocytosis (p = 0.0269), and juçara fruit-based product do not promote cytoprotection. At 23-week biliary hyperplasia continued present, and liver necrosis was evident in C group and CJ group. Hepatic degeneration was greater in C group, and megalocytosis was evident in the cancer-induced groups, without cytoprotection by juçara fruit-based product. In kidney, at 23-week, renal congestion was more evident in CJ group, and tubular degeneration in C and CE groups. Important hepatic and renal injuries were observed in rats induced to colorectal cancer and the supplementation with juçara fruit-based product, in the dose used, did not interfere in the prevention and resolution of these injuries, mainly with the chronic use