Carcinoma hepatocelular metastático em Amazona aestiva / Metastatic hepatocellular carcinoma in Amazona aestiva

Background: Hepatocellular carcinoma is a primary malignant tumor of the liver tissue and its occurrence in birds is considered rare. The tumor can occur as a single mass leading to hepatomegaly, or as multiple nodules in the liver. In animals of the genus Amazona, only 1 case of metastatic hepatocellular carcinoma has been reported in the United States, therefore, little is known about its epidemiology and clinicopathological aspects in these species. In this context, the aim of this work was to describe a case of metastatic hepatocellular carcinoma in an Amazona aestiva. Case: A blue-fronted amazon parrot (Amazona aestiva) was referred to necropsy after being found dead in its enclosure. On examination, it presented cachectic body score. Examination of the coelomic cavity, revealed a serous translucent fluid and adhesions between the liver and peritoneum.A red mass restricted to the right hepatic lobe and raised to the capsular surface, interspersed with whitish and dark red multifocal areas was observed. When cut, this mass was soft, protruding, multilobulated, whitish and with a friable reddish center. Additionally, on the dorsal surface of the left lung lobe, there was a rounded, well defined, whitish, and soft nodule. Microscopically, partial replacement of the hepatic parenchyma was observed by neoplastic proliferation of cuboidal epithelial cells, organized in mantle and supported by a scarce fibrovascular stroma. Cells have large, eosinophilic, well-delimited cytoplasm, with a central, oval nucleus, loose chromatin, and evident nucleolus. Moderate pleomorphism was characterized by anisocytosis, anisokaryosis, and aberrant nuclei. In the lung, a focally extensive mass with a pattern similar to that seen in the liver was observed. In the kidney, multifocal neoplastic emboli were noted. Liver immunohistochemistry was performed. Positive and negative controls were used to validate the reaction; however, there was no immunolabelling for the evaluated antibodies. Discussion: The histopathological characteristics observed in this study favored the diagnosis of hepatocellular carcinoma (HCC) with metastasis to kidney and lung. Primary liver tumors are rare in wild birds. In ducks, experimental studies have pointed aflatoxins and the duck hepatitis B virus as oncogenic agents, however, in birds of the genus Amazona, there are no studies that evaluate predisposing factors to the development of liver carcinoma. Macroscopically, hepatocellular carcinoma may present in massive, nodular or diffuse forms. In birds, the right lobe is the largest, which may suggest that this lobe is more prone to the development of HCC, as seen in the present case. The solid form, similar to that observed in this report, seems to be more commonly observed, as seen in the wild bird reports consulted. Metastases most often spread hematogenous, and in the present report there was metastasis to kidneys and lungs, which is a common feature for this neoplasm. In the present case, there was no labeling by any of the antibodies, perhaps because of their aggressiveness, associated with autolytic factors that prevent the labeling of antibodies, in addition to the specificity in the antibodyantigen relationship. This tumor must be differentiated from other liver tumors such as cholangiocarcinoma, and also the well-differentiated hepatocellular adenoma, in addition to non-neoplastic conditions. HCC should be considered as a differential diagnosis for Amazona aestiva found dead in the enclosure without previous clinical signs. This neoplasm is rare in Amazon parrots and reports should be encouraged in order to contribute to the understanding of the epidemiological and clinicopathological aspects of the tumor.

Clinical and pathological aspects of progressive hyperphosphatasemia associated with vacuolar hepatopathy and hepatocellular carcinoma in a Scottish Terrier bitch / Aspectos clínicos e patológicos da hiperfosfatasemia progressiva associada à hepatopatia vacuolar e carcinoma hepatocelular em uma cadela da raça Scottish Terrier

Hyperphosphatasemia refers to an increase in alkaline phosphatase serum activity, and Scottish Terriers (STs) are predisposed to develop this condition of uncertain pathogenesis. This study describes a case of progressive hyperphosphatasemia with vacuolar hepatopathy and hepatocellular carcinoma (HCC) in a ST bitch. This dog had a five-year clinical follow-up with progressive hyperphosphatasemia (up to 5503 U/L) and with ultrasound findings and histologic diagnosis of vacuolar hepatopathy, in addition to posterior onset of HCC. A steroidogenic adrenal panel revealed an increase of adrenocortical hormones, especially progesterone and androstenedione, consistent with a subdiagnosed hypercortisolism. Euthanasia was elected and at necropsy, multinodular, yellow to red masses were observed in the liver, which were histologically and immunohistochemically defined as HCC. The association of the clinical, imaging, biochemical, adrenal panel and pathologic findings allowed to characterize and confirm a progressive disorder in this ST bitch associated with elevated adrenocortical hormones.

Hiperfosfatasemia é o aumento sérico de fosfatase alcalina, sendo que Scorrish Terriers estão predispostos a desenvolverem essa condição de patogênese desconhecida. Este trabalho descreve um caso de hiperfosfatasemia progressiva com hepatopatia vacuolar e carcinoma hepatocelular em um canino da raça Scottish Terrier. Uma cadela Scottish Terrier foi acompanhada clinicamente por cinco anos devido à hiperfosfatasemia persistente (até 5503 U/L), com achados ultrassonográficos e histológicos compatíveis com hepatopatia vacuolar, além de posterior desenvolvimento de carcinoma hepatocelular. O painel esteroidogênico realizado indicou aumento dos hormônios adrenocorticais, principalmente progesterona e androstenediona, consistente com diagnóstico de hipercortisolismo subdiagnosticado "atípico". Devido ao prognóstico desfavorável, a eutanásia foi realizada e na necropsia, massas amarelas a vermelhas e multinodulares foram observadas no fígado, com diagnóstico de carcinoma hepatocelular pela análise histológica e imuno-histoquímica. A associação dos achados clínicos, de imagem, bioquímicos, do painel androgênico e patológicos permitiram caracterizar e confirmar um distúrbio progressivo no canino da raça Scottish Terrier associado ao aumento dos hormônios adrenocorticais.

Metastatic intranasal mastocytoma in a dog

Background: Mast cell tumors (MCTs) are neoplasms originating from mast cells, which can be well or poorly differentiated. They are considered the most commonly diagnosed malignant cutaneous neoplasm in dogs; however, intranasal forms are still little reported. Thus, this study seeks to report a case of unilateral intranasal MCT exhibiting submandibular lymph node metastasis. Case: A 11-year-old-and-4-month-old dog of undefined breed (UB), weighing 41 kg, was referred to the Veterinary Medical Teaching Hospital of the University of Passo Fundo (UPF), in the state of Rio Grande do Sul, Brazil. Presenting a clinical history of bilateral purulent nasal secretion, accompanied by sneezing in the two months prior to admission, in addition to vomiting and diarrhea. Auxiliary tests were requested, including skull X-ray, cytology of the nasal cavity with a swab, and collection of material from the submandibular lymph node directly through cytology with a needle. Cytological findings from the right nasal cavity were consistent with mast cell tumors (MCTs). Cytological analysis of the left nasal cavity was compatible with dysplasia/cellular reactivity. A heterogeneous population of cells was detected on cytology of the right submandibular lymph node. These findings were consistent with MCT lymph node metastasis. Skull radiography showed an increase in both opacity and soft tissue extension, surpassing the palate, from the canine tooth through the caudal region of the maxillary sinuses to the last molar, without bone destruction. The dog was then admitted for an abdominal ultrasound, which showed no changes in the spleen or liver. The leukocyte count showed mild lymphopenia and the presence of reactive lymphocytes. Through the buffy coat, the presence of rare round cells, compatible with circulating mast cells, was detected. Due to the biological behavior of the neoplasm and its anatomical location, the established therapy was based on the use of vinblastine and prednisolone. The patient did not show any clinical improvements. In a joint decision with the patient's guardian, the dog was euthanized. Discussion: Intranasal MCTs commonly present progressive and intermittent unilateral epitaxis, mucopurulent nasal discharge, dyspnea, and ocular discharge. Several anatomical sites were associated with more aggressive neoplastic phenotypes; those with an unfavorable prognosis were mainly those present in the oral and intranasal mucosa. Cytopathological examination is considered a highly sensitive method for the diagnosis of MCTs. Metastases are present in more than 90% of mucosal MCTs, usually affecting regional lymph nodes and associated with a poor prognosis. Radiography is considered a useful test in determining the size and location of tumors in the nasal cavity. Chemotherapy plays an important role in the treatment, especially in cases like the one described in this report, in which surgical excision is not possible due to the anatomical location of the neoplasm. Intranasal MCTs are uncommon in dogs. In this case, he presented aggressive, metastatic behavior and a poor response to antineoplastic therapy. Furthermore, due to the location of these tumors, they may be clinically similar to a number of other upper respiratory tract diseases, posing a diagnostic challenge. Therefore, it is essential that the search for differential diagnoses be carried out through auxiliary tests, such as cytology and imaging.

Extraskeletal osteosarcoma in the duodenum of a dog

Background: Osteosarcoma is the most observed primary bone tumor in dogs, and may affect the appendicular and axial skeletons. In addition, it may be present in extraskeletal form, accounting for only 1% of cases. As shown by few reports in the literature, the involvement of the intestinal region by is rare. The objective of this study was to report the case of a 13-year-old Yorkshire dog, submitted to an exploratory laparotomy for suspected partial intestinal obstruction, diagnosed with extraskeletal osteosarcoma. Case: A 13-year-old dog, Yorkshire Terrier, male, presented clinical signs of gastrointestinal abnormalities. An ultrasound examination was performed and was found a mass in small intestine region with wall and lumen invasion. Then, was realized exploratory laparotomy and detected intestinal obstruction due to a mass with approximately 5.0 x 6.0 x 4.4 cm localized in duodenum. Surgical removal was performed and the sample sent to the veterinary diagnostic laboratory for histopathological examination. The sample had an irregular surface and firm consistency. In addition, when cut, the mass enveloped the intestinal layers and sometimes obstructed the lumen. Then, the sample were processed routinely for histopathology. After that, in microscopy evaluation was detected cell proliferation, affecting all layers of intestine. In detail, cells were elongated with pleomorphism marked and atypical mitosis. In addition, there was production of cartilage and bone matrix. So, due the absence of others sites, the neoplasm was considered primary of intestine. After that, to evaluate the expression of KI-67 and COX-2 was performed, and the cell proliferation index was 54.0% and the COX-2 expression was moderate in less than 10% of neoplastic cells. After the surgery, the patient was hospitalized for a week and continue the treatment in home. Afterwards, the tutor received the diagnosis, but even though he was instructed on the severity of the case, he chose not to undergo chemotherapy. After three months, the patient presented abdominal fluid and nodules in your liver, suggesting metastasis, but without diagnosis confirmation. The patient died five months after the diagnosis of extraskeletal osteosarcoma. However, no necropsy was realized, impossibility the diagnosis confirm. Discussion: The frequency of extraskeletal osteosarcoma in dogs remains unknown, with the mammary glands being the most affected site. In the present study, osteosarcoma affects the duodenal region and no reports of this neoplasm in the duodenum of dogs have been found in the literature. The clinical sign of dyschezia was important for the tutor to refer the animal to the veterinarian and perform the ultrasound in an attempt to elucidate the case, as the tumor mass is not always palpable. Histopathological examination and immunohistochemistry were necessary for the differential diagnosis and to establish the prognosis, although after the surgery the tutor chose not to perform chemotherapy. Extraskeletal osteosarcoma are usually highly metastatic, mainly affecting the lymph nodes and liver. In this case, the patient presented a liver nodule three months after the tumor removal surgery, but unfortunately, there was no diagnostic confirmation. Such neoplastic type is rarer and more aggressive than appendicular and axial osteosarcoma, with an average survival of 1 to 3 months. In this case, as a necropsy was not obtained, we cannot attribute the survival time to the disease. The survival rates of osteosarcomas in dogs are few months, but in the present case, although the patient died five months after surgery, the failure to perform a necropsy compromises the attribution of survival time to extraskeletal osteosarcoma.

Cutaneous epitheliotropic lymphoma in a Lhasa Apso

Background: Lymphoma neoplasms originate from the lymphocytes. Anatomically, these tumors can be classified into multicentric, digestive, mediastinal, and cutaneous forms. The etiology of cutaneous lymphoma remains unclear; however, it has been associated with chronic skin inflammation. The definitive diagnosis is based on histological analysis and immunohistochemistry, although fine-needle aspiration cytology has shown good results. The aim of this paper is to describe the clinicopathological aspects of a case of cutaneous epitheliotropic T cell lymphoma, classified as mycosis fungoides, in a Lhasa Apso dog. Case: A 8-year-old bitch Lhasa Apso with multiple non-pruritic skin nodules and history of 10-day evolution was referred to the Veterinary Hospital of the Centro Universitário do Espírito Santo (UNESC), Colatina, ES, Brazil. The nodules were erythematous, exophytic, firm, circumscribed, and measured 0.2-4 cm in diameter in locations throughout the animal's body. An incisional biopsy was performed with an 8-mm punch and sent for histopathological examination. An infiltrative, poorly demarcated, non-encapsulated, densely cellular neoplasm, which was replacing the dermal collagen and displacing the adnexa, was observed in the dermis. The tumor was composed of a population of round cells, with generally distinct cell borders and a small-to-moderate amount of eosinophilic cytoplasm. The nuclei were irregularly rounded and occasionally edentulous, with vesicular chromatin, a visible nucleus, and 11 mitotic figures in an area of 2.37 mm2 . The immunohistochemical test, which was positive for the CD3 marker, confirmed the diagnosis of T cell lymphoma. On an ultrasound to identify metastasis, the liver showed heterogeneous parenchyma, heterogeneous expansive formation, areas of cavitary appearance, and cytology compatible with lymphoma. Antineoplastic chemotherapy was administered using the CHOP regimen (cyclophosphamide, doxorubicin, vincristine, and prednisone). However, the animal died after 45 days. Discussion: A diagnosis of the mycosis fungoides type of cutaneous epitheliotropic T cell lymphoma was established based on clinical, laboratory, anatomopathological, and immunohistochemical findings. Pruritus is a common clinical condition in animals with mycosis fungoides, particularly in those with the erythrodermic form of the disease. Epitheliotropic lymphomas have no sexual or racial predilections and usually affect dogs over 9 years of age. The Cocker Spaniel, English Bulldog, Boxer, Golden Retriever, Scottish Terrier, Briard, English Springer Spaniel, Beagle, German Shepherd, and English Cocker Spaniel breeds are frequently affected by these lymphomas. These neoplasms can have a primary skin origin, or they can be secondary and associated with lymphoma found elsewhere in the body. Chemotherapy is the treatment of choice, especially in cases with multifocal distribution. Protocol preference varies with disease stage, patient clinical and laboratory conditions, and the degree of toxicity. Commonly used chemotherapy regimens include L-CHOP (vincristine, cyclophosphamide, doxorubicin, L-asparaginase, and prednisolone), CHOP, COP (cyclophosphamide, vincristine, and prednisone), LAP (lomustine, L-asparaginase, and prednisolone), LOPP (lomustine, vincristine, procarbazine, prednisolone), chlorambucil, and prednisolone. The prognosis of canine epitheliotropic cutaneous lymphoma is unfavorable, with a survival time ranging from a few months to 2 years. The animal in this study survived for 105 days. In addition, epitheliotropic cutaneous T cell lymphoma is aggressive, which may result in a shorter survival time in animals affected by this type of tumor.

Cytotoxicity of isolated compounds from Picrasma crenata (Vell) Engl. in animal tumor cell (HTC) / Citotoxicidade de compostos isolados de Picrasma crenata (Vell) Engl. em células tumorais animais (HTC)

The study aim was to evaluate the cytotoxic activity, using the MTT test [3-(4,5-Dimethilthiazol-2-yl)-2,5-diphenil tetrazolium bromide], from the crude extract of Picrasma crenata (Pau Tenente) and its isolated compounds, quassin and parain, in culture of rat liver tumor cells (HTC). The test was carried out exposing the cells for 24, 48 and 72 hours to concentrations of 5, 10, 50, 100, 200, 300, 400, 500 and 1000 µg of crude extract of Pau Tenente/mL of culture medium and 1, 5, 10, 15, 20, 40, 60, 80 and 100 µg of quassin or parain compounds/mL of culture medium. The absorbances averages results obtained showed that the crude extract did not present cytotoxicity for the HTC cells in all the concentrations and evaluated times. For quassin, the concentrations of 80 and 100 µg/mL were cytotoxic, after 72 hours of treatment. For parain, the concentrations of 1, 5, 20, 40, 60, 80 and 100 µg/mL, in 72 hours, were cytotoxic, revealing a new activity for this compound. Thus, the results demonstrate a first indication of the cytotoxic activity of compounds quassin and parain, adding an important social and economic value to them, and may have application in future research and in pharmaceutical industry.

O objetivo do estudo foi avaliar a atividade citotóxica, por meio do teste MTT [brometo de 3-(4,5-dimetiltiazol-2-il)-2,5-difenil tetrazólio], do extrato bruto de Picrasma crenata (Pau Tenente) e seus compostos isolados, quassina e paraína, em cultura de células tumorais de fígado de rato (HTC). O teste foi realizado expondo as células por 24, 48 e 72 horas às concentrações de 5, 10, 50, 100, 200, 300, 400, 500 e 1000 µg de extrato bruto de Pau Tenente/mL de meio de cultura e 1, 5, 10, 15, 20, 40, 60, 80 e 100 µg de compostos de quassina ou paraína/mL de meio de cultura. Os resultados das médias de absorbâncias obtidos mostraram que o extrato bruto não apresentou citotoxicidade para as células HTC em todas as concentrações e tempos avaliados. Para quassina, as concentrações de 80 e 100 µg/mL foram citotóxicas, após 72 horas de tratamento. Para a paraína, as concentrações de 1, 5, 20, 40, 60, 80 e 100 µg/mL, em 72 horas, foram citotóxicas, revelando uma nova atividade para este composto. Assim, os resultados demonstram uma primeira indicação da atividade citotóxica dos compostos quassina e paraína, agregando-lhes importante valor social e econômico, podendo ter aplicação em pesquisas futuras e na indústria farmacêutica.

Explore the antiproliferative phytocompounds from ethanolic extracts of Citrus paradisi against liver cancer cell line by chemical analysis using TLC and FT-IR spectroscopy / Explorar os fitocompostos antiproliferativos de extratos etanólicos de Citrus paradisi contra linhagem de células de câncer de fígado por análise química usando espectroscopia de TLC e FT-IR

The aim of the present study was to evaluate the in vitro antiproliferative activity of ethanolic extract of leaves and fruits Citrus paradisi plant on HepG-2 liver cell lines by MTT (3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2Hterazolium bromide) assay and to isolate and characterize the antiproliferative compounds by TLC (Thin layer chromatography) and FT-IR (Fourier transforms Infrared) spectroscopy. Qualitative phytochemical screening tests were performed to detect phytochemicals compounds from the crude extracts. Antioxidant activity of the plant extracts were characterized by using DPPH (2,2-Diphenyl-1-picrylhydrazyl) free radical scavenging method. The results showed that antioxidant activity using DPPH were found to be increased in a concentration dependent manner and decreased cell viability and cell growth inhibition in a dose dependent manner. The findings from this study indicated that fruit extract exhibited good antiproliferation and antioxidant potential. The seven functional groups of phytocompounds such as carboxylic acid, amine salt, aromatic compounds, cyclic alkene, aldehyde, fluoro compounds and alkene were detected by FT-IR which indicated that fruit extracts of Citrus paradisi possessed vast potential as a medicinal drug especially in liver cancer treatment.

O objetivo do presente estudo foi avaliar a atividade antiproliferativa in vitro do extrato etanólico de folhas e frutos da planta Citrus paradisi em linhagens de células hepáticas HepG-2 por MTT (3- (4, 5-dimetil-2-tiazolil) -2, Ensaio de brometo de 5-difenil-2H-terazólio) e isolar e caracterizar os compostos antiproliferativos por espectroscopia de TLC (cromatografia de camada fina) e FT-IR (infravermelho com transformadas de Fourier). Testes qualitativos de triagem fitoquímica foram realizados para detectar compostos fitoquímicos nos extratos brutos. A atividade antioxidante dos extratos vegetais foi caracterizada pelo método de eliminação de radicais livres DPPH (2,2-difenil-1-picrilhidrazil). Os resultados mostraram que a atividade antioxidante usando DPPH aumentou de uma maneira dependente da concentração e diminuiu a viabilidade celular e a inibição do crescimento celular de uma maneira dependente da dose. Os resultados deste estudo indicaram que o extrato de fruta exibiu bom potencial antiproliferação e antioxidante. Os sete grupos funcionais de fitocompostos, como ácido carboxílico, sal de amina, compostos aromáticos, alceno cíclico, aldeído, compostos de flúor e alceno, foram detectados por FT-IR, o que indicou que extratos de frutas de Citrus paradisi possuíam vasto potencial como medicamento, especialmente no tratamento de câncer do fígado.

Lobectomia total como tratamento de hemangiossarcoma hepático em um cão - relato de caso / Total lobectomy as treatment of hepatic hemangiosarcoma in a dog - case report / Lobectomía total como tratamiento de hemangiosarcoma hepático en un perro - reporte de caso

O hemangiossarcoma é uma neoplasia mesenquimal maligna agressiva com elevada taxa de morbidade e de mortalidade em cães e gatos; que se desenvolve mais frequentemente em baço, fígado, coração, ossos além de poder manifestar metástases regionais. O presente relato tem por objetivo descrever um caso de um canino diagnosticado com hemangiossarcoma hepático em lobo caudado submetido a uma lobectomia total. A técnica cirúrgica consistiu na utilização da sutura de guilhotina modificada na base do lobo acometido utilizando fio de polidioxanona e, com o auxílio de um bisturi elétrico unipolar, e respeitando o distanciamento de 0,5 cm da sutura, foi realizada a lobectomia. Após a remoção do lobo, foi fixada uma esponja hemostática de colágeno na região da incisão como forma de auxílio no controle hemorrágico. Foi indicado também a realização de tratamento quimioterápico adjuvante, entretanto o tutor do animal optou pela não realização da mesma. Ainda assim, o paciente do presente relato obteve uma boa resposta ao procedimento, e o tutor ficou muito satisfeito, relatando que houve uma grande melhora na qualidade de vida do animal e que ele não sentia mais dor, voltando a ter o seu comportamento habitual.(AU)

Hemangiosarcoma is an aggressive malignant mesenchymal neoplasm with a high rate of morbidity and mortality in dogs and cats; which develops more frequently in the spleen, liver, heart, bones, in addition to being able to manifest regional metastases. The present report aims to describe a case of a canine diagnosed with hepatic hemangiosarcoma in the caudate lobe submitted to a total lobectomy. The surgical technique consisted of using a modified guillotine suture at the base of the affected lobe using polydioxanone thread and, with the aid of a unipolar electric scalpel, and respecting the distance of 0.5 cm from the suture, lobectomy was performed. After removing the lobe, a hemostatic collagen sponge was fixed in the region of the incision as an aid in hemorrhagic control. Adjuvant chemotherapy treatment was also indicated, however the animal's tutor chose not to perform it. Even so, the patient in the present report had a good response to the procedure, and the tutor was very satisfied, reporting that there was a great improvement in the animal's quality of life and that he no longer felt pain, returning to his usual behavior.(AU)

El hemangiosarcoma es una neoplasia mesenquimatosa maligna agresiva con una alta tasa de morbilidad y mortalidad en perros y gatos; la cual se desarrolla con mayor frecuencia en bazo, hígado, corazón, huesos, además de poder manifestar metástasis regionales. El presente reporte tiene como objetivo describir un caso de un canino con diagnóstico de hemangiosarcoma hepático en el lóbulo caudado sometido a una lobectomía total. La técnica quirúrgica consistió en utilizar una sutura de guillotina modificada en la base del lóbulo afectado con hilo de polidioxanona y, con la ayuda de un bisturí eléctrico unipolar, y respetando la distancia de 0,5 cm de la sutura, se realizó la lobectomía. Después de retirar el lóbulo, se fijó una esponja hemostática de colágeno en la región de la incisión como ayuda para el control hemorrágico. También se indicó tratamiento de quimioterapia adyuvante, sin embargo el tutor del animal optó por no realizarlo. Aun así, el paciente del presente reporte tuvo una buena respuesta al procedimiento, y el tutor quedó muy satisfecho, informando que hubo una gran mejoría en la calidad de vida del animal y que ya no sintió dolor, volviendo a su comportamiento habitual.(AU)

Explore the antiproliferative phytocompounds from ethanolic extracts of Citrus paradisi against liver cancer cell line by chemical analysis using TLC and FT-IR spectroscopy

Abstract The aim of the present study was to evaluate the in vitro antiproliferative activity of ethanolic extract of leaves and fruits Citrus paradisi plant on HepG-2 liver cell lines by MTT (3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-terazolium bromide) assay and to isolate and characterize the antiproliferative compounds by TLC (Thin layer chromatography) and FT-IR (Fourier transforms Infrared) spectroscopy. Qualitative phytochemical screening tests were performed to detect phytochemicals compounds from the crude extracts. Antioxidant activity of the plant extracts were characterized by using DPPH (2,2-Diphenyl-1-picrylhydrazyl) free radical scavenging method. The results showed that antioxidant activity using DPPH were found to be increased in a concentration dependent manner and decreased cell viability and cell growth inhibition in a dose dependent manner. The findings from this study indicated that fruit extract exhibited good antiproliferation and antioxidant potential. The seven functional groups of phytocompounds such as carboxylic acid, amine salt, aromatic compounds, cyclic alkene, aldehyde, fluoro compounds and alkene were detected by FT-IR which indicated that fruit extracts of Citrus paradisi possessed vast potential as a medicinal drug especially in liver cancer treatment.

Resumo O objetivo do presente estudo foi avaliar a atividade antiproliferativa in vitro do extrato etanólico de folhas e frutos da planta Citrus paradisi em linhagens de células hepáticas HepG-2 por MTT (3- (4, 5-dimetil-2-tiazolil) -2, Ensaio de brometo de 5-difenil-2H-terazólio) e isolar e caracterizar os compostos antiproliferativos por espectroscopia de TLC (cromatografia de camada fina) e FT-IR (infravermelho com transformadas de Fourier). Testes qualitativos de triagem fitoquímica foram realizados para detectar compostos fitoquímicos nos extratos brutos. A atividade antioxidante dos extratos vegetais foi caracterizada pelo método de eliminação de radicais livres DPPH (2,2-difenil-1-picrilhidrazil). Os resultados mostraram que a atividade antioxidante usando DPPH aumentou de uma maneira dependente da concentração e diminuiu a viabilidade celular e a inibição do crescimento celular de uma maneira dependente da dose. Os resultados deste estudo indicaram que o extrato de fruta exibiu bom potencial antiproliferação e antioxidante. Os sete grupos funcionais de fitocompostos, como ácido carboxílico, sal de amina, compostos aromáticos, alceno cíclico, aldeído, compostos de flúor e alceno, foram detectados por FT-IR, o que indicou que extratos de frutas de Citrus paradisi possuíam vasto potencial como medicamento, especialmente no tratamento de câncer do fígado.

Biliary cystadenoma in a lion (Panthera leo)

Animals in human care often exceed the life expectancy of animals in the wild when they have veterinarian follow-ups, no competition for space or food, and continuous care. Advanced age favors the development of mutations that often trigger cancer, which sometimes causes death. There are reports in the literature on neoplasms in lions in the liver, which are one of the main organs affected. A 20-year-old lion specimen was received for necropsy at the Federal University of Paraná, Curitiba, Brazil. Multiple cystic dilations were observed in the liver. Histologically, they were internally covered by a simple cuboidal epithelium, similar to that observed in the bile ducts, without cellular atypia. The adjacent liver parenchyma presented with mild disorganization of the hepatocyte cords. Biliary cystadenomas are benign growths formed by a thin opaque capsule filled with a slightly yellowish translucent fluid compressing the adjacent liver parenchyma, as highlighted in this case. The epithelium of the cysts was positive for anti-cytokeratin (CK) (EIA/A3E) and anti-CK7, confirming histogenesis in the bile ducts. This study reports a case of biliary cystadenoma in a Panthera leo specimen.(AU)

Cytotoxicity of extracts and compounds isolated from Croton echioides in animal tumor cell (HTC) / Citotoxicidade de extratos e compostos isolados de Croton echioides em célula tumoral animal (HTC)

The search for compounds with anticancer effects is of paramount importance today due to the high incidence of the disease. The Euphorbiaceae family is known for having compounds with therapeutic properties, one of its genera being Croton. It has several species, which contain compounds already known for their biological activities, presenting anti-inflammatory, antimicrobial and anticancer properties. Thus, the cytotoxicity/antiproliferative activity of semi-purified fractions and compounds isolated from Croton echioides in liver tumor cells of Rattus norvegicus (HTC) was evaluated by the MTT test. The semi-purified fractions showed cytotoxicity at concentrations above 200 µg/mL, at 24, 48 and 72 hours, reaching cell viability of 24.78% [400 µg/mL] at 24 hours, 12.79% [500 µg/mL] at 48 hours and 10.57% [300 µg/mL] at 72 hours. For the isolated compounds, lupeol had a cytotoxic effect in all concentrations (1, 5, 10, 15, 20, 40, 60, 80 and 100 µg/mL) and tested times (24, 48 and 72 hours), reaching minimum viability of 4.37% [100 µg/mL], within 72 hours. The clerodan diterpenes CEH-1 and CEH-4 also showed antiproliferative activity, with minimum viability of 36.19% [100 µg/mL] over 72 hours and 21.33% [100 µg/mL] over 48 hours, respectively. However, the clerodan diterpenes CEH-2 and CEH-3 did not shows a cytotoxic effect for HTC cells. Thus, there is a cytotoxic/antiproliferative potential of C. echioides against tumor cells, with targeted to mitochondrial enzymes, associated with cell proliferation, indicating that this species deserves prominence in the search for new molecules for the treatment of cancer.

A busca por compostos com efeitos anticâncer é de suma importância nos dias atuais devido à alta incidência desta doença. A família Euphorbiaceae é conhecida por possuir compostos com propriedades terapêuticas, sendo um de seus gêneros o Croton. Este possui diversas espécies, que contêm compostos já conhecidos por suas atividades biológicas, apresentando propriedades anti-inflamatórias, antimicrobianas e anticancerígenas. Assim, a citotoxicidade/atividade antiproliferativa de frações semipurificadas e compostos isolados de Croton echioides em células tumorais hepáticas de Rattus norvegicus (HTC) foi avaliada pelo teste MTT. As frações semipurificadas apresentaram citotoxicidade em concentrações acima de 200 µg/mL, em 24, 48 e 72 horas, atingindo viabilidade celular de 24,78% [400 µg/mL] em 24 horas, 12,79% [500 µg/mL] em 48 horas e 10,57% [300 µg/mL] às 72 horas. Para os compostos isolados, o lupeol teve efeito citotóxico em todas as concentrações (1, 5, 10, 15, 20, 40, 60, 80 e 100 µg/mL) e tempos testados (24, 48 e 72 horas), atingindo a viabilidade mínima de 4,37% [100 µg/mL], em 72 horas. Os diterpenos clerodan CEH-1 e CEH-4 também apresentaram atividade antiproliferativa, com viabilidade mínima de 36,19% [100 µg/mL] em 72 horas e 21,33% [100 µg/mL] em 48 horas, respectivamente. No entanto, os diterpenos clerodanos CEH-2 e CEH-3 não apresentaram efeito citotóxico para células HTC. Assim, existe um potencial citotóxico/antiproliferativo de C. echioides contra células tumorais, com enzimas direcionadas a enzimas mitocondriais, associadas à proliferação celular, indicando que esta espécie merece destaque na busca de novas moléculas para o tratamento do câncer.

Virucidal activity of oriental hornet Vespa orientalis venom against hepatitis C virus

Background Hepatitis C virus (HCV) infection is a major worldwide health problem that can cause liver fibrosis and hepatocellular carcinoma (HCC). The clinical treatment of HCV infection mainly relies on the use of direct-acting antivirals (DAAs) that are usually expensive and have side effects. Therefore, achieving the discovery of more successful agents is always urgent. In this context, antiviral compounds that inhibit viral infections and disease progression with important therapeutic activities have been identified in animal venoms including arthropod toxins. This indicates that arthropod venoms represent a good natural source of promising candidates for new antivirals. Methods The antiviral activity of the wasp venom (WV), isolated from the Oriental hornet (Vespa orientalis), was assessed using cell culture technique with human hepatocellular carcinoma-derived cell line (Huh7it-1) and the recombinant strain of HCV genotype 2a (JFH1). Results The results revealed that WV inhibited HCV infectivity with 50% inhibitory concentration (IC50) of 10 ng/mL, while the 50% cytotoxic concentration (CC50) was 11,000 ng/mL. Time of addition experiment showed that the WV blocked HCV attachment/entry to the cells probably through virucidal effect. On the other hand, the venom showed no inhibitory effect on HCV replication. Conclusion WV can inhibit the entry stage of HCV infection at non-cytotoxic concentrations. Therefore, it could be considered a potential candidate for characterization of natural anti-HCV agents targeting the entry step.(AU)

Biological compatibility of oxidized cellulose vs. porcine gelatin to control bleeding in liver lesions in rats

ABSTRACT Purpose: To compare biological compatibility, hemostasis, and adhesion formation between oxidized regenerated cellulose and lyophilized hydrolyzed porcine collagen in liver trauma. Methods: Forty male Wistar rats constituted two groups: group A (oxidized cellulose) and group B (lyophilized hydrolyzed collagen). Standardized liver trauma was made, and the hemostatic agent was applied. Animals in subgroups A7 and B7 were submitted to euthanasia and relaparotomy after seven days, and in subgroups A14 and B14 after 14 days. Macroscopic and microscopic results were evaluated. Results: There was no fluid in the cavity in any of the animals, and adhesions were present in all of them. In the analysis after seven days, the adhesions were grades 3 or 4 and consisted of omentum, small intestine, and abdominal wall (p<0.05). In both groups, the mesh was surrounded by a capsule, which was not observed after 14 days. In the evaluation after 14 days, adhesions were grades 2 or 3 (p>0.05). The microscopic examination showed subacute and chronic reactions, in both groups and in both timepoints, with similar frequency. The intensity of fibrosis always presented positive scores. Microabscesses and xanthomatous macrophages were observed in both groups. Conclusions: There was no superiority of one agent over the other.

A case of metastatic non-teratoid ocular medulloepithelioma in an adult horse

This case reports an eight-years-old horse that presented a unilateral tumor in the left ocular orbital region diagnosed on the basis of clinical signs, gross anatomopathological and histopathological investigations. The affected area was exenteration and submitted to histopathological evaluation and was diagnosed as a non-teratoid medulloepithelioma. This is a rare ocular neoplasm that usually affects young individuals – and was composed by neuroepithelium arranged in tubules and Flexner-Wintersteiner rosettes without presence of heteroplastic components. The animals died and was submitted to post-mortem evaluation, revealing presence of medulloepithelioma metastases at the internal region of the head, lymph nodes, lungs, and liver. This neoplasm usually presents a benign frame; however, the case herein described shows that it can be highly malignant and aggressive since it can cause extensive lesions and spread metastases to visceral organs.

A case of metastatic non-teratoid ocular medulloepithelioma in an adult horse

This case reports an eight-years-old horse that presented a unilateral tumor in the left ocular orbital region diagnosed on the basis of clinical signs, gross anatomopathological and histopathological investigations. The affected area was exenteration and submitted to histopathological evaluation and was diagnosed as a non-teratoid medulloepithelioma. This is a rare ocular neoplasm that usually affects young individuals and was composed by neuroepithelium arranged in tubules and Flexner-Wintersteiner rosettes without presence of heteroplastic components. The animals died and was submitted to post-mortem evaluation, revealing presence of medulloepithelioma metastases at the internal region of the head, lymph nodes, lungs, and liver. This neoplasm usually presents a benign frame; however, the case herein described shows that it can be highly malignant and aggressive since it can cause extensive lesions and spread metastases to visceral organs.(AU)

Non-epitheliotropic cutaneous lymphoma with systemic dissemination in a dog

Background: Cutaneous lymphoma is a highly malignant neoplasm, which can originate in the epidermis or dermis, aswell as be disseminated to other organs such as lung, heart, arm, liver and bone marrow. It comes in the form of nodes ofvarious sizes, erythematous and alopecic, itching may or may not occur. The diagnosis is made by cytological and histopathological examination of the compromised tissue. However, it is important to perform complementary tests for clinicalstaging and prognostic characterization. The objective is to report a case of non-epitheliotropic cutaneous lymphoma withsystemic dissemination in a dog.Case: A female mixed bred adult canine was attended at the Veterinary Hospital of the Federal University of Mato Grossodo Sul with a history of progressive weight loss and the presence of alopecic and non-pruritic subcutaneous nodules in thetorso, nasal plane and pelvic limbs, starting 4 months ago. After approximately 20 days, the presence of rapidly evolvingulcerated nodules was noted. On physical examination, generalized lymphadenomegaly was observed and among thedermatological findings were multiple nodules of varying sizes with the presence of ulceration in the center of the lesions,alopecia, erythema and raised edges, in the region of the nasal sinus, pelvic and thoracic limbs, tail, thoracolumbar andabdominal region. The animal also presented right pelvic limb edema with painful sensibility to manipulation CBC andbiochemical tests (albumin, alanine aminotransferase, creatinine, urea, alkaline phosphatase, globulins, total proteins andfractions) were performed, being observed as normocytic normochromic type anemia alteration (erythrocytes: 2.78 106/µL; hemoglobin: 6.8 g/µL; globular volume: 18.8%), leukopenia (4,000/mm3) with presence of metamyelocytes (120/mm3) and rods (1,080/mm3) and lymphopenia (80/mm3). Three samples of the nodules were...

Non-epitheliotropic cutaneous lymphoma with systemic dissemination in a dog

Background: Cutaneous lymphoma is a highly malignant neoplasm, which can originate in the epidermis or dermis, aswell as be disseminated to other organs such as lung, heart, arm, liver and bone marrow. It comes in the form of nodes ofvarious sizes, erythematous and alopecic, itching may or may not occur. The diagnosis is made by cytological and histopathological examination of the compromised tissue. However, it is important to perform complementary tests for clinicalstaging and prognostic characterization. The objective is to report a case of non-epitheliotropic cutaneous lymphoma withsystemic dissemination in a dog.Case: A female mixed bred adult canine was attended at the Veterinary Hospital of the Federal University of Mato Grossodo Sul with a history of progressive weight loss and the presence of alopecic and non-pruritic subcutaneous nodules in thetorso, nasal plane and pelvic limbs, starting 4 months ago. After approximately 20 days, the presence of rapidly evolvingulcerated nodules was noted. On physical examination, generalized lymphadenomegaly was observed and among thedermatological findings were multiple nodules of varying sizes with the presence of ulceration in the center of the lesions,alopecia, erythema and raised edges, in the region of the nasal sinus, pelvic and thoracic limbs, tail, thoracolumbar andabdominal region. The animal also presented right pelvic limb edema with painful sensibility to manipulation CBC andbiochemical tests (albumin, alanine aminotransferase, creatinine, urea, alkaline phosphatase, globulins, total proteins andfractions) were performed, being observed as normocytic normochromic type anemia alteration (erythrocytes: 2.78 106/µL; hemoglobin: 6.8 g/µL; globular volume: 18.8%), leukopenia (4,000/mm3) with presence of metamyelocytes (120/mm3) and rods (1,080/mm3) and lymphopenia (80/mm3). Three samples of the nodules were...(AU)

Evaluation of potential hepatotoxicity induced by Bortezomib

Bortezomib, an inhibitor of 26S proteasome, is an anti-cancer therapeutic agent used in different cancer types. It leads to the arrest of the cancerous cell cycle by inhibiting angiogenesis and inducing apoptosis. Liver is the vital organ for detoxification and excretion of toxic products. The treatment with chemotherapy is a challenge, drugs are used to destroy cancer cells, but healthy cells can be affected during cancer treatment as well. The main objective of this study was to analyze the histopathological and biochemical effects of bortezomib on liver. Twenty-four female C57BL/6 mice were distributed into 4 groups, bortezomib injected treatment groups (Btz1, Btz2) and saline injected control groups (C1, C2). Bortezomib and saline were treated twice per week for 6 weeks and sacrificed at the end of one day (Btz1, C1) and 4 weeks (Btz2, C2) after the last injection. Liver samples were examined for histopathological analysis and the serum samples processed for biochemical analysis. Tissue samples were fixed, routinely processed, sectioned, and stained with Hematoxylin and Eosin (H&E). Periodic Acid-Schiff (PAS), Sudan Black staining and Masson's trichrome histochemical staining methods were performed to characterize the lesions. Histopathological analysis of the Btz1 and Btz2 groups revealed acute hepatic morphological changes such as hepatocellular swelling (cloudy swelling), necro-inflammatory reaction, and increased mononuclear polyploidy. Based on the negative staining with PAS and Sudan Black staining, hepatocellular swelling was diagnosed as hydropic degeneration. Necro-inflammatory reaction observed in the form of acute hepatitis was composed of mainly mononuclear cell infiltration accompanied by multifocal necrotic foci. Kupffer cell proliferation was observed in parallel with degenerative and necrotic changes. An Increase in hepatocellular mononuclear polyploidy visualized as hepatocytes with a single enlarged nucleus was detected in all liver sections of Btz1 and Btz2 groups. Individual cases of cholestasis (n = 1) and mild hepatic fibrosis (n = 1) were also reported. Significant elevated levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT) were detected in bortezomib treated groups. Few clinical cases reported liver injury related to bortezomib used for cancer treatment. However, the liver was not considered as a target for bortezomib treatment. Our data suggesting that bortezomib caused liver damage and induces elevations in serum levels. The reported hepatic lesions including hepatocellular swelling, acute hepatitis and mononuclear polyploidy were mainly mild and moderate in severity. The increase of polyploidy in liver tissue of mice treated with bortezomib in this study was explained as a reaction of the liver facing the drug-induced hepatic damage. The mechanism leading to the hepatotoxicity of bortezomib treatment is not known but the production of a toxic metabolite through its metabolism in the liver can be suggested. Moreover, no recovery was also observed in histopathological and biochemical analyses suggesting that the bortezomib effect is non-reversible four weeks after the drug was withdrawn. Patients should be informed about the possibility of acute drug-induced hepatitis and hepatotoxicity of this chemotherapeutic agent after the treatment.(AU)

Tamponamento cardíaco secundário ao hemangiossarcoma em cães / Cardiac tamponade secondary to hemangiosarcoma in a dog

Background: Nonspecific clinical manifestations such as apathy, anorexia and diarrhea are common in the clinical routine, and therefore may mask the severity of its triggering factor. When patients presenting this symptomatology are referred to the care center, it is essential that a thorough investigation is performed to clarify the primary causes of these manifestations, and for this, complementary imaging exams may be necessary. The objective of this study is to describe the clinical and imaging aspects of a canine with cardiac hemangiosarcoma and to correlate with the pathophysiology of the alterations observed, in order to optimize the clinical care of patients with nonspecific clinical signs and affected by this alteration. Case: A 10-year-old Pitbull dog was treated with a history of vomiting, loss of appetite and diarrhea. Upon clinical examination, the animal presented dehydration level and distended abdomen. On ultrasound examination, hepatomegaly was observed, associated with signs of double layer in the gallbladder wall and the presence of moderate peritoneal effusion. On chest radiography, the cardiac silhouette showed an important increase in size with loss of shape and contours. Dorsal displacement of the trachea, greater contact of the heart with the sternum and displacement of the caudal vena cava were observed. In addition, an amorphous and poorly defined structure with radiopacity of soft tissues was observed in the region of the atrium and right ventricle, causing the obliteration of pulmonary fields. On echocardiographic examination, a significant amount of free anechogenic fluid was observed in the pericardial sac, confirming the suspicion of pericardial effusion, with consequent collapse of the wall of the right heart chambers during systole. Furthermore, an amorphous, poorly defined and heterogeneous structure was observed in the right atrium wall. Due to the location of the neoformation, the patient's general clinical condition, euthanasia was chosen. During the necropsy, the ocular, oral and preputial mucosae were moderately pale. In the abdomen, the presence of hydroperitoneum and hepatomegaly was observed. In the thoracic cavity, hemothorax, enlarged heart were identified and, in the right atrium, a reddish mass was identified. In addition, the lungs were whitish and hypercrepitating at the edges of the cranial lobes; the rest of the organ was moderately hyperemic, hypocrepitating. In histopathological examination of the liver, the centrilobular region showed chronic passive congestion associated with necrosis and multifocal degeneration of hepatocytes. Neoplastic proliferation of mesenchymal cells, moderately cellular, non-encapsulated, of infiltrative growth was observed in the heart. Neoplastic cells formed bundles, with a tendency to organize themselves into small blood vessels filled with red blood cells. The cytoplasm was moderate, elongated, indistinct, eosinophilic and homogeneous. The nucleus was large, unique, elongated, with finely dotted chromatin and sometimes with one or two nucleoli evident. Anisocytosis, anisocariosis and cell pleomorphism were moderate. Interwoven with neoplastic cells, a moderate presence of multifocal lymphohistioplasmocytic inflammatory infiltrate was observed. The definitive diagnosis of cardiac changes was hemangiosarcoma. Discussion: Hemangiosarcoma is a vascular endothelial cell neoplasm with high metastatic power and unfavorable prognosis. When located in the heart, it is commonly found in the auricle and right atrium and the cardiovascular changes caused by this neoplasm as well as the severity of these changes vary according to size and location. Clinical manifestations can be quite nonspecific and are usually associated with hemodynamic impairment, causing signs of right or left congestive heart failure.

Prognostic impact of alterations in E-cadherin and P-cadherin expression in canine mammary tumors

A expressão de caderinas tem sido correlacionada ao desenvolvimento e agressividade de neoplasias epiteliais, entretanto, em tumores mamários caninos, seu prognóstico de importância é incerto. Devido a esse fato, a expressão das moléculas de adesão intracelular E e P-caderina e a correlação com a sobrevida global foram analisadas em 25 tumores de glândula mamária canina. A redução da expressão da caderina-E foi correlacionada com o tipo histológico, alto grau histológico e taxa de sobrevida global. A coloração de P-caderina foi maior em tumores malignos, sem relação com outras características clínico-patológicas de agressividade. Os resultados deste estudo sugerem uma relação entre menor expressão de E e P-caderina e pior prognóstico em tumores mamários caninos, incluindo menor sobrevida global.