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.

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

ABSTRACT: 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.

RESUMO: 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.

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.

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)

Antioxidant, genotoxic, antigenotoxic, and antineoplastic activities of apitoxin produced by Apis mellifera in Northeast, Brazil / Atividades antioxidantes, genotóxicas, antigenotóxicas e antineoplásicas da apitoxina produzida por Apis mellifera no Nordeste do Brasil

The objective was to evaluate the in vitro antioxidant, genotoxic, antigenotoxic, and antineoplastic activities of apitoxin produced by the bee Apis mellifera. The antioxidant activity of the apitoxin solution was evaluated using the DPPH (2,2-diphenyl-1-picrilhydrazyl) method. Genotoxic potential of apitoxin was analyzed by comparing the mean DNA damage indices (idDNA) of L929 strain fibroblasts exposed to hydrogen peroxide (H2O2 - genotoxic substance), distilled water, or apitoxin. The antigenotoxic effect of apitoxin was analyzed by assessing the percentage decrease in H2O2-induced genotoxicity in L929 fibroblasts co-treated with three concentrations of the aqueous apitoxin solution and subjected to comet assay. In vitro antineoplastic activity in human tumor cell lines of prostate adenocarcinoma (PC3), hepatocellular carcinoma (HEPGE2), melanoma (MAD-MB435), and astrocytoma (SNB19), were verified by MTT [3- (4) bromide colorimetric method, 5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium]. Apitoxin had no genotoxic effect on L929 cells at concentrations of 30, 10, and 5 µg/mL after 24 hours of exposure. This effect was only evident at 50 µg/mL. Apitoxin promoted a significant reduction in DNA damage index (idDNA) at all concentrations tested. At 30 µg/mL, apitoxin attenuated the genotoxic effects induced by H2O2. Apitoxin also demonstrated in vitro antineoplastic potential, since the cytotoxic effect was observed at concentrations of 50 µg/mL and 25 µg/mL, with significant reduction in viability percentage of PC3 tumor cell lines, HEPGE2, MAD-MB435, and SNB19. The high antioxidant activity associated with the absence of genotoxic effect and the genoprotective and antineoplastic effect demonstrated by apitoxin here provide indications of apitoxins therapeutic potential.(AU)

O objetivo deste estudo foi avaliar as atividades antioxidantes, genotóxicas, antigenotóxicas e antineoplásicas in vitro da apitoxina produzida pela abelha Apis mellifera. A atividade antioxidante da solução da apitoxina foi avaliada pelo método DPPH (2,2-difenil-1-picrilhidrazil). O potencial genotóxico da apitoxina foi analisado através dos índices médios de dano ao DNA (idDNA) dos fibroblastos da linhagem L929 expostos à peróxido de hidrogênio (H2O2 - substância genotóxica), água destilada ou apitoxina. O efeito antigenotóxico da apitoxina foi analisado através da avaliação da diminuição percentual na genotoxicidade induzida por H2O2 nos fibroblastos L929 co-tratados com três concentrações da solução aquosa de apitoxina e submetidos ao ensaio cometa. A atividade antineoplásica in vitro em linhagens celulares tumorais humanas de adenocarcinoma da próstata (PC3), carcinoma hepatocelular (HEPGE2), melanoma (MAD-MB435) e astrocitoma (SNB19), foram verificadas pelo método colorimétrico do brometo de MTT [3- (4), 5-dimetiltiazol -2-il) -2,5-difeniltetrazólio]. A apitoxina não teve efeito genotóxico nas células L929 nas concentrações de 30, 10 e 5 µg / mL após 24 horas de exposição. Este efeito foi apenas evidente a 50 µg / mL. A apitoxina promoveu uma redução significativa no índice de danos ao DNA (idDNA) em todas as concentrações testadas. A 30 µg / mL, a apitoxina atenuou os efeitos genotóxicos induzidos por H2O2. A apitoxina também demonstrou potencial antineoplásico in vitro, uma vez que o efeito citotóxico foi observado em concentrações de 50 µg / mL e 25 µg / mL, com redução significativa na porcentagem de viabilidade das linhagens celulares de PC3, HEPGE2, MAD-MB435 e SNB19. A alta atividade antioxidante associada à ausência de efeito genotóxico e o efeito genoprotetor e antineoplásico demonstrado pela apitoxina aqui fornecem indicações do potencial terapêutico da apitoxina.(AU)

Pathological and immunohistochemical aspects of primary hepatobiliary neoplasms in cats

ABSTRACT: Primary hepatobiliary neoplasms (PHN) are uncommon in cats, and originate in hepatocytes, intra- and extrahepatic bile ducts, mesenchymal cells, and cells of neuroendocrine origin. The aim of this study was to determine the frequency of PHN in cats diagnosed in the metropolitan region of Porto Alegre (RS), Brazil, for a period of 17 years, determining their epidemiological, anatomopathological and immunohistochemical aspects. Necropsy reports of 2.090 cats were analyzed, 125 were diagnosed with primary hepatobiliary diseases, of which 15 were cases of PHN, representing 12% of the specific hepatobiliary conditions and 0.7% of the necropsies. All PHN were malignant, of which 93.3% had epithelial origin and 6.7% presented mesenchymal origin. Cholangiocarcinoma was the most commonly diagnosed neoplasm, followed by hepatocellular carcinoma and hemangiosarcoma. In general, cats with no defined breed were the most affected. Concerning sex, 60% were females and 40% males. Age ranged from five to 18 years, with a mean age of 10.5 years (median of ten years). Grossly, cholangiocarcinoma and hemangiosarcoma were multinodular and hepatocellular carcinoma was massive. Microscopically, cholangiocarcinomas were arranged in acini and ducts, whereas hepatocellular carcinomas were arranged in solid sheets or trabeculae. On immunohistochemistry, cholangiocarcinomas, hepatocellular carcinomas, and hemangiosarcomas were positive for the antibodies CK 7, Hep Par-1, and vimentin and von Willebrand factor, respectively.

RESUMO: Neoplasias hepatobiliares primárias (NHP) são incomuns em gatos e se originam de hepatócitos, células dos ductos biliares intra e extra-hepáticos, células mesenquimais e ainda células de origem neuroendócrina. O objetivo do trabalho foi determinar a frequência das NHP em gatos diagnosticados na Região Metropolitana de Porto Alegre, no período de 17 anos, abordando seus aspectos epidemiológicos, anatomopatológicos e imuno-histoquímicos (IHQ). Foram analisados os laudos de necropsia de 2.090 gatos sendo que 125 foram diagnosticados com doenças hepatobiliares primárias, destes 15 foram casos de NHP, representando 12% das condições hepatobiliares específicas e 0,7% do total de necropsias. Todos os diagnósticos de NHP eram malignos, destes 93,3% apresentaram origem epitelial e 6,7% mesenquimal. Colangiocarcinoma foi a neoplasia mais diagnosticada, seguido do carcinoma hepatocelular e hemangiossarcoma. De uma maneira geral, os gatos sem raça definida foram os mais acometidos. Em relação ao sexo 60% eram fêmeas e 40% machos. A idade variou de cinco a 18 anos, com a idade média de 10,5 anos (mediana de 10 anos). Macroscopicamente o colangiocarcinoma e hemangiossarcoma eram multinodulares, e o carcinoma hepatocelular, maciço. À histologia, houve predomínio do arranjo acinar e ductal nos colangiocarcinomas e sólido, no carcinoma hepatocelular. Na IHQ os colangiocarcinomas foram reativos para CK 7, carcinoma hepatocelular para Hep Par-1 e hemangiossarcoma para vimentina e fator de von Willebrand.

Pathological and immunohistochemical aspects of primary hepatobiliary neoplasms in cats / Aspectos patológicos e imuno-histoquímicos de neoplasias hepatobiliares primárias em gatos

Primary hepatobiliary neoplasms (PHN) are uncommon in cats, and originate in hepatocytes, intra- and extrahepatic bile ducts, mesenchymal cells, and cells of neuroendocrine origin. The aim of this study was to determine the frequency of PHN in cats diagnosed in the metropolitan region of Porto Alegre (RS), Brazil, for a period of 17 years, determining their epidemiological, anatomopathological and immunohistochemical aspects. Necropsy reports of 2.090 cats were analyzed, 125 were diagnosed with primary hepatobiliary diseases, of which 15 were cases of PHN, representing 12% of the specific hepatobiliary conditions and 0.7% of the necropsies. All PHN were malignant, of which 93.3% had epithelial origin and 6.7% presented mesenchymal origin. Cholangiocarcinoma was the most commonly diagnosed neoplasm, followed by hepatocellular carcinoma and hemangiosarcoma. In general, cats with no defined breed were the most affected. Concerning sex, 60% were females and 40% males. Age ranged from five to 18 years, with a mean age of 10.5 years (median of ten years). Grossly, cholangiocarcinoma and hemangiosarcoma were multinodular and hepatocellular carcinoma was massive. Microscopically, cholangiocarcinomas were arranged in acini and ducts, whereas hepatocellular carcinomas were arranged in solid sheets or trabeculae. On immunohistochemistry, cholangiocarcinomas, hepatocellular carcinomas, and hemangiosarcomas were positive for the antibodies CK 7, Hep Par-1, and vimentin and von Willebrand factor, respectively.(AU)

Neoplasias hepatobiliares primárias (NHP) são incomuns em gatos e se originam de hepatócitos, células dos ductos biliares intra e extra-hepáticos, células mesenquimais e ainda células de origem neuroendócrina. O objetivo do trabalho foi determinar a frequência das NHP em gatos diagnosticados na Região Metropolitana de Porto Alegre, no período de 17 anos, abordando seus aspectos epidemiológicos, anatomopatológicos e imuno-histoquímicos (IHQ). Foram analisados os laudos de necropsia de 2.090 gatos sendo que 125 foram diagnosticados com doenças hepatobiliares primárias, destes 15 foram casos de NHP, representando 12% das condições hepatobiliares específicas e 0,7% do total de necropsias. Todos os diagnósticos de NHP eram malignos, destes 93,3% apresentaram origem epitelial e 6,7% mesenquimal. Colangiocarcinoma foi a neoplasia mais diagnosticada, seguido do carcinoma hepatocelular e hemangiossarcoma. De uma maneira geral, os gatos sem raça definida foram os mais acometidos. Em relação ao sexo 60% eram fêmeas e 40% machos. A idade variou de cinco a 18 anos, com a idade média de 10,5 anos (mediana de 10 anos). Macroscopicamente o colangiocarcinoma e hemangiossarcoma eram multinodulares, e o carcinoma hepatocelular, maciço. À histologia, houve predomínio do arranjo acinar e ductal nos colangiocarcinomas e sólido, no carcinoma hepatocelular. Na IHQ os colangiocarcinomas foram reativos para CK 7, carcinoma hepatocelular para Hep Par-1 e hemangiossarcoma para vimentina e fator de von Willebrand.(AU)

Pathological and immunohistochemical aspects of primary hepatobiliary neoplasms in cats / Aspectos patológicos e imuno-histoquímicos de neoplasias hepatobiliares primárias em gatos

Primary hepatobiliary neoplasms (PHN) are uncommon in cats, and originate in hepatocytes, intra- and extrahepatic bile ducts, mesenchymal cells, and cells of neuroendocrine origin. The aim of this study was to determine the frequency of PHN in cats diagnosed in the metropolitan region of Porto Alegre (RS), Brazil, for a period of 17 years, determining their epidemiological, anatomopathological and immunohistochemical aspects. Necropsy reports of 2.090 cats were analyzed, 125 were diagnosed with primary hepatobiliary diseases, of which 15 were cases of PHN, representing 12% of the specific hepatobiliary conditions and 0.7% of the necropsies. All PHN were malignant, of which 93.3% had epithelial origin and 6.7% presented mesenchymal origin. Cholangiocarcinoma was the most commonly diagnosed neoplasm, followed by hepatocellular carcinoma and hemangiosarcoma. In general, cats with no defined breed were the most affected. Concerning sex, 60% were females and 40% males. Age ranged from five to 18 years, with a mean age of 10.5 years (median of ten years). Grossly, cholangiocarcinoma and hemangiosarcoma were multinodular and hepatocellular carcinoma was massive. Microscopically, cholangiocarcinomas were arranged in acini and ducts, whereas hepatocellular carcinomas were arranged in solid sheets or trabeculae. On immunohistochemistry, cholangiocarcinomas, hepatocellular carcinomas, and hemangiosarcomas were positive for the antibodies CK 7, Hep Par-1, and vimentin and von Willebrand factor, respectively.(AU)

Neoplasias hepatobiliares primárias (NHP) são incomuns em gatos e se originam de hepatócitos, células dos ductos biliares intra e extra-hepáticos, células mesenquimais e ainda células de origem neuroendócrina. O objetivo do trabalho foi determinar a frequência das NHP em gatos diagnosticados na Região Metropolitana de Porto Alegre, no período de 17 anos, abordando seus aspectos epidemiológicos, anatomopatológicos e imuno-histoquímicos (IHQ). Foram analisados os laudos de necropsia de 2.090 gatos sendo que 125 foram diagnosticados com doenças hepatobiliares primárias, destes 15 foram casos de NHP, representando 12% das condições hepatobiliares específicas e 0,7% do total de necropsias. Todos os diagnósticos de NHP eram malignos, destes 93,3% apresentaram origem epitelial e 6,7% mesenquimal. Colangiocarcinoma foi a neoplasia mais diagnosticada, seguido do carcinoma hepatocelular e hemangiossarcoma. De uma maneira geral, os gatos sem raça definida foram os mais acometidos. Em relação ao sexo 60% eram fêmeas e 40% machos. A idade variou de cinco a 18 anos, com a idade média de 10,5 anos (mediana de 10 anos). Macroscopicamente o colangiocarcinoma e hemangiossarcoma eram multinodulares, e o carcinoma hepatocelular, maciço. À histologia, houve predomínio do arranjo acinar e ductal nos colangiocarcinomas e sólido, no carcinoma hepatocelular. Na IHQ os colangiocarcinomas foram reativos para CK 7, carcinoma hepatocelular para Hep Par-1 e hemangiossarcoma para vimentina e fator de von Willebrand.(AU)

Primary hepatic neoplasms in cattle / Neoplasmas hepáticos primários de bovinos

Primary hepatic neoplasms are mostly detected in cattle as incidental findings in slaughterhouses or diagnosed at the necropsy, wherein it may be related to the cause of death. A proper characterization of primary hepatic neoplasms is essential to provide an accurate diagnosis, especially at the slaughter lines, in order to reduce erroneous condemnations. This work aimed to characterize the gross, histological, and immunohistochemical features of primary liver neoplasms detected in slaughtered cattle in Southern Brazil. Nineteen primary hepatic neoplasms were identified. Grossly, these lesions were classified according to their distribution, as focal, multifocal, or diffuse. Histologically, the shape and arrangement of the cells, as well as possible malignant features were evaluated. Immunohistochemistry (IHC) was also performed for biliary epithelium (anti-CK7) and hepatocytes (anti-Hep Par-1) markers. Hepatocellular carcinoma (84.2%) was the most frequently detected hepatic neoplasm, followed by cholangiocarcinoma (15.8%), and these were only identified in adult cows. Hepatocellular carcinomas occurred as solitary masses or multifocal nodules, which on the cut surface were often green. Cholangiocarcinomas occurred as multifocal nodules, occasionally showing an umbilicated appearance. Histologically, hepatocellular carcinomas had mostly trabecular and solid patterns, while cholangiocarcinomas presented mostly a solid arrangement. Upon IHC, all hepatocellular carcinomas were immunolabeled for anti-Hep Par-1, ranging from mild (25%), moderate (31.2%) to marked (43.7%), while immunolabeling for anti-CK7 was detected only in one case of cholangiocarcinoma.(AU)

Os neoplasmas hepáticos primários são detectados em bovinos principalmente como achados incidentais em matadouros ou diagnosticados na necropsia, quando podem estar relacionados à causa da morte. A caracterização adequada dos tumores hepáticos primários é essencial para obter diagnósticos precisos, especialmente nas linhas de abate, com o propósito de reduzir condenações errôneas. Este trabalho teve o objetivo de determinar as características macroscópicas, histológicas e imuno-histoquímicas dos neoplasmas primários do fígado de bovinos abatidos em um matadouro-frigorífico no Sul do Brasil. Dezenove neoplasias hepáticas primárias foram identificadas. Macroscopicamente, os tumores hepáticos foram classificados de acordo com sua distribuição, como focais, multifocais ou difusos. Histologicamente, a forma e o arranjo das células e possíveis características malignas foram avaliados. Também foi realizada imuno-histoquímica (IHQ) para marcadores de epitélio biliar (anti-CK7) e hepatócitos (anti-Hep Par-1). O carcinoma hepatocelular (84,2%) foi o neoplasma hepático mais frequentemente detectado, seguido pelo colangiocarcinoma (15,8%). Esses tumores foram identificados apenas em vacas adultas. Os carcinomas hepatocelulares eram vistos como massas solitárias ou nódulos multifocais que na superfície de corte geralmente eram esverdeados. Os colangiocarcinomas foram observados como nódulos multifocais, ocasionalmente com aspecto umbilicado. Histologicamente, os padrões mais observados nos carcinomas hepatocelulares foram trabeculares e sólidos, enquanto nos colangiocarcinomas o arranjo sólido foi o mais frequente. Na IHQ, todos os carcinomas hepatocelulares foram marcados por anti-Hep Par-1, com marcação que variou de leve (25%), moderada (31,2%) a acentuada (43,7%); imunomarcação para anti-CK7 foi detectada em apenas um caso de colangiocarcinoma.(AU)

Primary hepatic neoplasms in cattle / Neoplasmas hepáticos primários de bovinos

Primary hepatic neoplasms are mostly detected in cattle as incidental findings in slaughterhouses or diagnosed at the necropsy, wherein it may be related to the cause of death. A proper characterization of primary hepatic neoplasms is essential to provide an accurate diagnosis, especially at the slaughter lines, in order to reduce erroneous condemnations. This work aimed to characterize the gross, histological, and immunohistochemical features of primary liver neoplasms detected in slaughtered cattle in Southern Brazil. Nineteen primary hepatic neoplasms were identified. Grossly, these lesions were classified according to their distribution, as focal, multifocal, or diffuse. Histologically, the shape and arrangement of the cells, as well as possible malignant features were evaluated. Immunohistochemistry (IHC) was also performed for biliary epithelium (anti-CK7) and hepatocytes (anti-Hep Par-1) markers. Hepatocellular carcinoma (84.2%) was the most frequently detected hepatic neoplasm, followed by cholangiocarcinoma (15.8%), and these were only identified in adult cows. Hepatocellular carcinomas occurred as solitary masses or multifocal nodules, which on the cut surface were often green. Cholangiocarcinomas occurred as multifocal nodules, occasionally showing an umbilicated appearance. Histologically, hepatocellular carcinomas had mostly trabecular and solid patterns, while cholangiocarcinomas presented mostly a solid arrangement. Upon IHC, all hepatocellular carcinomas were immunolabeled for anti-Hep Par-1, ranging from mild (25%), moderate (31.2%) to marked (43.7%), while immunolabeling for anti-CK7 was detected only in one case of cholangiocarcinoma.(AU)

Os neoplasmas hepáticos primários são detectados em bovinos principalmente como achados incidentais em matadouros ou diagnosticados na necropsia, quando podem estar relacionados à causa da morte. A caracterização adequada dos tumores hepáticos primários é essencial para obter diagnósticos precisos, especialmente nas linhas de abate, com o propósito de reduzir condenações errôneas. Este trabalho teve o objetivo de determinar as características macroscópicas, histológicas e imuno-histoquímicas dos neoplasmas primários do fígado de bovinos abatidos em um matadouro-frigorífico no Sul do Brasil. Dezenove neoplasias hepáticas primárias foram identificadas. Macroscopicamente, os tumores hepáticos foram classificados de acordo com sua distribuição, como focais, multifocais ou difusos. Histologicamente, a forma e o arranjo das células e possíveis características malignas foram avaliados. Também foi realizada imuno-histoquímica (IHQ) para marcadores de epitélio biliar (anti-CK7) e hepatócitos (anti-Hep Par-1). O carcinoma hepatocelular (84,2%) foi o neoplasma hepático mais frequentemente detectado, seguido pelo colangiocarcinoma (15,8%). Esses tumores foram identificados apenas em vacas adultas. Os carcinomas hepatocelulares eram vistos como massas solitárias ou nódulos multifocais que na superfície de corte geralmente eram esverdeados. Os colangiocarcinomas foram observados como nódulos multifocais, ocasionalmente com aspecto umbilicado. Histologicamente, os padrões mais observados nos carcinomas hepatocelulares foram trabeculares e sólidos, enquanto nos colangiocarcinomas o arranjo sólido foi o mais frequente. Na IHQ, todos os carcinomas hepatocelulares foram marcados por anti-Hep Par-1, com marcação que variou de leve (25%), moderada (31,2%) a acentuada (43,7%); imunomarcação para anti-CK7 foi detectada em apenas um caso de colangiocarcinoma.(AU)

In vivo induction of hepatocellular carcinoma by diethylnitrosoamine and pharmacological intervention in Balb C mice using Bergenia ciliata extracts / Indução in vivo de carcinoma hepatocelular por dietilnitrosoamina e intervenção farmacológica em camundongos balb c usando extratos de Bergenia ciliata

Background Hepatocellular carcinoma is the most frequent primary malignancy of liver and accounts for as many as one million deaths worldwide in a year. Objectives The aim of the present study was to evaluate the anti-cancerous efficiency of Bergenia ciliata rhizome against diethylnitrosoamine induced hepatocarcinogenesis in Balb C mice. Methods One percent diethylnitrosoamine was prepared by using 99 ml of normal saline NaCl (0.9 percent) solution to which was added 1 ml of concentrated diethylnitrosoamine (DEN) solution (0.01 g/l). Extract of Bergenia ciliata was prepared by maceration technique. Mice were classified into four groups as follows: Group 1 a control group (N=7) received saline solution (3.5 l/mg), group 2 (N=14) received diethylnitrosoamine (3.5 l/mg) intraperitoneally once in a week for eight consecutive weeks, group 3 (N=7) received plant extract (150 mg/kg (Body weight)) once in a week, while group 4 (N=7) was given combination of diethylnitrosoamine (3.5 l/mg) and plant extract (150 mg/kg (Body weight)). After eight weeks of DEN induction group 2 mice were divided into two subgroups containing seven mice each, subgroup 1 was sacrificed while subgroup 2 was treated with plant extract (150 mg/kg (Body weight)) once in a week for eight consecutive weeks. Results The model of DEN injected hepatocellular carcinomic (HCC) mice elicited significant decline in levels of albumin with concomitant significant elevations in tumor markers aspartate aminotransferase, alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alpha feto protein (AFP), gamma glutamyl transferase (Y-GT), 5 nucleotidase (5NT), glucose-6-phosphate dehydrogenase (G6PDH) and bilirubin. The intraperitoneal administration of B. ciliata as a protective agent, produced significant increase in albumin levels with significant decrease in the levels of tumor markers aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alpha feto protein (AFP), gamma glutamyl transferase (Y-GT), 5 nucleotidase (5NT), glucose-6-phosphate dehydrogenase (G6PDH) and bilirubin. Conclusion Bergenia ciliata has potent antioxidant activity, radical scavenging capacity and anticancerous properties. Bergenia ciliata extracts may provide a basis for development of anti-cancerous drug.(AU)

Antecedentes O carcinoma hepatocelular é a neoplasia primária mais frequente do fígado e é responsável por até um milhão de mortes em todo o mundo em um ano. Objetivos O objetivo do presente estudo foi avaliar a eficiência anticancerígena do rizoma de Bergenia ciliata contra a hepatocarcinogênese induzida por dietilnitrosoamina em camundongos balb c. Métodos Um por cento de dietilnitrosoamina foi preparado usando 99 ml de solução salina normal (0,9 por cento) à qual foi adicionado 1 ml de solução concentrada de dietilnitrosoamina (DEN) (0,01 g / l). O extrato de Bergenia ciliata foi preparado pela técnica de maceração. Os ratos foram classificados em quatro grupos: Grupo 1 grupo controle (N = 7) recebeu solução salina (3,5 mL / mg), grupo 2 (N = 14) recebeu dietilnitrosoamina (3,5 mL / mg) por via intraperitoneal uma vez por semana para oito semanas consecutivas, o grupo 3 (N = 7) recebeu extrato vegetal (150 mg / kg (peso corporal)) uma vez por semana, enquanto o grupo 4 (N = 7) recebeu combinação de dietilnitrosoamina (3,5 l / mg) e extrato (150 mg / kg (peso corporal). Após oito semanas do grupo de indução DEN 2 ratos foram divididos em dois subgrupos contendo sete ratos cada, subgrupo 1 foi sacrificado enquanto subgrupo 2 foi tratado com extrato vegetal (150 mg / kg)) uma vez por semana durante oito semanas consecutivas. Resultados O modelo de camundongos hepatocelulares carcinômicos (CHC) injetados com DEN provocou declínio significativo nos níveis de albumina com elevações significativas concomitantes nos marcadores tumorais: aspartato aminotransferase, alanina aminotransferase (ALT), lactato desidrogenase (LDH), proteína alfa feto (AFP), gama glutamiltransferase (Y-GT), 5 nucleotidase (5NT), glicose-6-fosfato ehidrogenase (G6PDH) e bilirrubina. A administração intraperitoneal de B. ciliata como agente protetor produziu um aumento significativo nos níveis de albumina com uma diminuição significativa nos níveis dos marcadores tumorais: aspartato aminotransferase, alanina aminotransferase (ALT), lactato desidrogenase (LDH), proteína alfa feto (AFP), gama glutamiltransferase (Y-GT), 5 nucleotidase (5NT), glicose-6-fosfato desidrogenase (G6PDH) e bilirrubina. Conclusão Bergenia ciliata possui atividade antioxidante potente, capacidade de eliminação de radicais livres e propriedades anticancerígenas. Extratos de Bergenia ciliata podem fornecer uma base para o desenvolvimento de drogas anti-cancerígenas.(AU)

Clinical - pathological aspects of hepatocellular carcinoma in cattle

Background: The hepatocellular carcinoma (HCC) is a malignant neoplasm of hepatocytes of rare occurrence in farmanimals, with ruminants being the most affected species. This neoplasm is characterized by nonspecific symptoms and it isetiology in animals has not yet been fully elucidated, although aflatoxin has been shown to be a risk factor in the development this neoplasia. Since hepatic tumors in cattle are commonly incidental findings found in postmortem examination,the objective of this paper is to describe the clinical, laboratory and pathological findings in a cow with this neoplasm.Case: A 5-year-old, adult, Girolando cow weighing 350 kg was referred to the Veterinary Hospital of the Federal University of Recôncavo da Bahia, Brazil, with a history of decreased appetite and weakness for one month. The animal wasraised in a semi-intensive system, with a corn-based diet, and regular vaccination. Futhermore, was not treated at the farmof origin and three days before being admitted to the hospital, began to present edema of the dewlap. On examination atour center, the cow was in lean, active, with mucupurulent secretion in nostrils; ocular conjunctiva edema; and edema ofthe dewlap. The cow had neutrophilia, hypofibrinogemia, hypoproteinemia, and trombocytopenia. It also had tachycardia, tense abdomen, engorgement of the vessels of the face and jugular veins, stasis test and bilaterally positive jugularpulse. Although evidence of pain in reticulum was negative in the examination, the initial diagnostic suspicion establishedwas of traumatic reticular pericarditis. The therapeutic protocol instituted was daily monitoring, flunixin meglumine andflofernicol. The examination of rectal palpation revealed in the right flank an irregular-sized parenchymal structure withenlarged and palpacion in pain. Thus, by location and texture...

Clinical - pathological aspects of hepatocellular carcinoma in cattle

Background: The hepatocellular carcinoma (HCC) is a malignant neoplasm of hepatocytes of rare occurrence in farmanimals, with ruminants being the most affected species. This neoplasm is characterized by nonspecific symptoms and it isetiology in animals has not yet been fully elucidated, although aflatoxin has been shown to be a risk factor in the development this neoplasia. Since hepatic tumors in cattle are commonly incidental findings found in postmortem examination,the objective of this paper is to describe the clinical, laboratory and pathological findings in a cow with this neoplasm.Case: A 5-year-old, adult, Girolando cow weighing 350 kg was referred to the Veterinary Hospital of the Federal University of Recôncavo da Bahia, Brazil, with a history of decreased appetite and weakness for one month. The animal wasraised in a semi-intensive system, with a corn-based diet, and regular vaccination. Futhermore, was not treated at the farmof origin and three days before being admitted to the hospital, began to present edema of the dewlap. On examination atour center, the cow was in lean, active, with mucupurulent secretion in nostrils; ocular conjunctiva edema; and edema ofthe dewlap. The cow had neutrophilia, hypofibrinogemia, hypoproteinemia, and trombocytopenia. It also had tachycardia, tense abdomen, engorgement of the vessels of the face and jugular veins, stasis test and bilaterally positive jugularpulse. Although evidence of pain in reticulum was negative in the examination, the initial diagnostic suspicion establishedwas of traumatic reticular pericarditis. The therapeutic protocol instituted was daily monitoring, flunixin meglumine andflofernicol. The examination of rectal palpation revealed in the right flank an irregular-sized parenchymal structure withenlarged and palpacion in pain. Thus, by location and texture...(AU)

BjussuLAAO-II induces cytotoxicity and alters DNA methylation of cell-cycle genes in monocultured/co-cultured HepG2 cells

The use of animal venoms and their toxins as material sources for biotechnological applications has received much attention from the pharmaceutical industry. L-amino acid oxidases from snake venoms (SV-LAAOs) have demonstrated innumerous biological effects and pharmacological potential against different cancer types. Hepatocellular carcinoma has increased worldwide, and the aberrant DNA methylation of liver cells is a common mechanism to promote hepatic tumorigenesis. Moreover, tumor microenvironment plays a major role in neoplastic transformation. To elucidate the molecular mechanisms responsible for the cytotoxic effects of SV-LAAO in human cancer cells, this study aimed to evaluate the cytotoxicity and the alterations in DNA methylation profiler in the promoter regions of cell-cycle genes induced by BjussuLAAO-II, an LAAO from Bothrops jaracussu venom, in human hepatocellular carcinoma (HepG2) cells in monoculture and co-culture with endothelial (HUVEC) cells. Methods: BjussuLAAO-II concentrations were 0.25, 0.50, 1.00 and 5.00 μg/mL. Cell viability was assessed by MTT assay and DNA methylation of the promoter regions of 22 cell-cycle genes by EpiTect Methyl II PCR array. Results: BjussuLAAO-II decreased the cell viability of HepG2 cells in monoculture at all concentrations tested. In co-culture, 1.00 and 5.00 μg/mL induced cytotoxicity (p < 0.05). BjussuLAAO-II increased the methylation of CCND1 and decreased the methylation of CDKN1A in monoculture and GADD45A in both cell-culture models (p < 0.05). Conclusion: Data showed BjussuLAAO-II induced cytotoxicity and altered DNA methylation of the promoter regions of cell-cycle genes in HepG2 cells in monoculture and co-culture models. We suggested the analysis of DNA methylation profile of GADD45A as a potential biomarker of the cell cycle effects of BjussuLAAO-II in cancer cells. The tumor microenvironment should be considered to comprise part of biotechnological strategies during the development of snake-toxin-based novel drugs.(AU)

In vivo induction of hepatocellular carcinoma by diethylnitrosoamine and pharmacological intervention in Balb C mice using Bergenia ciliata extracts

Abstract Background Hepatocellular carcinoma is the most frequent primary malignancy of liver and accounts for as many as one million deaths worldwide in a year. Objectives The aim of the present study was to evaluate the anti-cancerous efficiency of Bergenia ciliata rhizome against diethylnitrosoamine induced hepatocarcinogenesis in Balb C mice. Methods One percent diethylnitrosoamine was prepared by using 99 ml of normal saline NaCl (0.9 percent) solution to which was added 1 ml of concentrated diethylnitrosoamine (DEN) solution (0.01 g/l). Extract of Bergenia ciliata was prepared by maceration technique. Mice were classified into four groups as follows: Group 1 a control group (N=7) received saline solution (3.5 l/mg), group 2 (N=14) received diethylnitrosoamine (3.5 l/mg) intraperitoneally once in a week for eight consecutive weeks, group 3 (N=7) received plant extract (150 mg/kg (Body weight)) once in a week, while group 4 (N=7) was given combination of diethylnitrosoamine (3.5 l/mg) and plant extract (150 mg/kg (Body weight)). After eight weeks of DEN induction group 2 mice were divided into two subgroups containing seven mice each, subgroup 1 was sacrificed while subgroup 2 was treated with plant extract (150 mg/kg (Body weight)) once in a week for eight consecutive weeks. Results The model of DEN injected hepatocellular carcinomic (HCC) mice elicited significant decline in levels of albumin with concomitant significant elevations in tumor markers aspartate aminotransferase, alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alpha feto protein (AFP), gamma glutamyl transferase (Y-GT), 5 nucleotidase (5NT), glucose-6-phosphate dehydrogenase (G6PDH) and bilirubin. The intraperitoneal administration of B. ciliata as a protective agent, produced significant increase in albumin levels with significant decrease in the levels of tumor markers aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alpha feto protein (AFP), gamma glutamyl transferase (Y-GT), 5 nucleotidase (5NT), glucose-6-phosphate dehydrogenase (G6PDH) and bilirubin. Conclusion Bergenia ciliata has potent antioxidant activity, radical scavenging capacity and anticancerous properties. Bergenia ciliata extracts may provide a basis for development of anti-cancerous drug.

Resumo Antecedentes O carcinoma hepatocelular é a neoplasia primária mais frequente do fígado e é responsável por até um milhão de mortes em todo o mundo em um ano. Objetivos O objetivo do presente estudo foi avaliar a eficiência anticancerígena do rizoma de Bergenia ciliata contra a hepatocarcinogênese induzida por dietilnitrosoamina em camundongos balb c. Métodos Um por cento de dietilnitrosoamina foi preparado usando 99 ml de solução salina normal (0,9 por cento) à qual foi adicionado 1 ml de solução concentrada de dietilnitrosoamina (DEN) (0,01 g / l). O extrato de Bergenia ciliata foi preparado pela técnica de maceração. Os ratos foram classificados em quatro grupos: Grupo 1 grupo controle (N = 7) recebeu solução salina (3,5 mL / mg), grupo 2 (N = 14) recebeu dietilnitrosoamina (3,5 mL / mg) por via intraperitoneal uma vez por semana para oito semanas consecutivas, o grupo 3 (N = 7) recebeu extrato vegetal (150 mg / kg (peso corporal)) uma vez por semana, enquanto o grupo 4 (N = 7) recebeu combinação de dietilnitrosoamina (3,5 l / mg) e extrato (150 mg / kg (peso corporal). Após oito semanas do grupo de indução DEN 2 ratos foram divididos em dois subgrupos contendo sete ratos cada, subgrupo 1 foi sacrificado enquanto subgrupo 2 foi tratado com extrato vegetal (150 mg / kg)) uma vez por semana durante oito semanas consecutivas. Resultados O modelo de camundongos hepatocelulares carcinômicos (CHC) injetados com DEN provocou declínio significativo nos níveis de albumina com elevações significativas concomitantes nos marcadores tumorais: aspartato aminotransferase, alanina aminotransferase (ALT), lactato desidrogenase (LDH), proteína alfa feto (AFP), gama glutamiltransferase (Y-GT), 5 nucleotidase (5NT), glicose-6-fosfato ehidrogenase (G6PDH) e bilirrubina. A administração intraperitoneal de B. ciliata como agente protetor produziu um aumento significativo nos níveis de albumina com uma diminuição significativa nos níveis dos marcadores tumorais: aspartato aminotransferase, alanina aminotransferase (ALT), lactato desidrogenase (LDH), proteína alfa feto (AFP), gama glutamiltransferase (Y-GT), 5 nucleotidase (5NT), glicose-6-fosfato desidrogenase (G6PDH) e bilirrubina. Conclusão Bergenia ciliata possui atividade antioxidante potente, capacidade de eliminação de radicais livres e propriedades anticancerígenas. Extratos de Bergenia ciliata podem fornecer uma base para o desenvolvimento de drogas anti-cancerígenas.

BjussuLAAO-II induces cytotoxicity and alters DNA methylation of cell-cycle genes in monocultured/co-cultured HepG2 cells

Background:The use of animal venoms and their toxins as material sources for biotechnological applications has received much attention from the pharmaceutical industry. L-amino acid oxidases from snake venoms (SV-LAAOs) have demonstrated innumerous biological effects and pharmacological potential against different cancer types. Hepatocellular carcinoma has increased worldwide, and the aberrant DNA methylation of liver cells is a common mechanism to promote hepatic tumorigenesis. Moreover, tumor microenvironment plays a major role in neoplastic transformation. To elucidate the molecular mechanisms responsible for the cytotoxic effects of SV-LAAO in human cancer cells, this study aimed to evaluate the cytotoxicity and the alterations in DNA methylation profiler in the promoter regions of cell-cycle genes induced by BjussuLAAO-II, an LAAO from Bothrops jaracussu venom, in human hepatocellular carcinoma (HepG2) cells in monoculture and co-culture with endothelial (HUVEC) cells.Methods:BjussuLAAO-II concentrations were 0.25, 0.50, 1.00 and 5.00 μg/mL. Cell viability was assessed by MTT assay and DNA methylation of the promoter regions of 22 cell-cycle genes by EpiTect Methyl II PCR array.Results:BjussuLAAO-II decreased the cell viability of HepG2 cells in monoculture at all concentrations tested. In co-culture, 1.00 and 5.00 μg/mL induced cytotoxicity (p < 0.05). BjussuLAAO-II increased the methylation of CCND1 and decreased the methylation of CDKN1A in monoculture and GADD45A in both cell-culture models (p < 0.05).Conclusion:Data showed BjussuLAAO-II induced cytotoxicity and altered DNA methylation of the promoter regions of cell-cycle genes in HepG2 cells in monoculture and co-culture models. We suggested the analysis of DNA methylation profile of GADD45A as a potential biomarker of the cell cycle effects of BjussuLAAO-II in cancer cells...(AU)

Hepatoprotective and antineoplastic potencial of red propolis produced by the bees Apis mellifera in the semiarid of Rio Grande do Norte, Brazil

ABSTRACT: The objective of this study was to evaluate the hepatoprotective effect of the honey bee Apis mellifera ethanolic extract of the red propolis, obtained in four municipalities of the Rio Grande do Norte semi-arid region, through an in vitro evaluation of the antineoplastic potential in human hepatic carcinoma (HepG2) and normal cell lines (L929), and from the comet assay in hepatic cell lines (ZF-L hepatocytes) to evaluate the genoprotective potential of the extract. The hepatoprotective effect was also evaluated in vivo by the induction of chronic experimental hepatic lesions in rodents (Rattus norvegicus Berkenhout, 1769), Wistar line, by intraperitoneal administration of thioacetamide (TAA) at the dose of 0.2g/kg. The animals were distributed in the following experimental groups: G1 (control), G2 (treated with 500mg/kg ethanolic extract of propolis), G3 (treated with 500mg/kg of ethanolic extract and TAA) and G4 (treated with TAA). All rats were submitted to serum biochemical, macroscopic, histological and stereological biochemical exams of the liver. It was verified the genoprotective effect of red propolis since the mean damages promoted to DNA in cells tested with the extract were significantly lower than the mean of the positive control damage (hydrogen peroxide). The red propolis extract did not present cytotoxic activity to the tumor cells of human liver cancer, as well as to normal ones. The absence of cytotoxicity in normal cells may indicate safety in the use of the propolis extract. The results of the serum biochemical evaluation showed that the serum levels of the aminotransferase enzymes (AST) did not differ significantly between G1, G2 and G3 when compared to each other. G4 showed significant increase in levels compared to the other groups, indicating that the administration of the extract did not cause liver toxicity, as well as exerted hepatoprotective effect against the hepatic damage induced by TAA. The G3 and G4 animals developed cirrhosis, but in G3 the livers were characterized by the presence of small regenerative nodules and level with the surface of the organ, whereas in G4 the livers showed large regenerative nodules. The livers of the G1 and G2 animals presented normal histological appearance, whereas the livers of the G3 animals showed regenerative nodules surrounded by thin septa of connective tissue, and in G4 the regenerative nodules were surrounded by thick septa fibrous connective tissue. The analysis of the hepatic tissues by means of stereology showed that there was no statistical difference between the percentage of hepatocytes, sinusoids, and collagens in G1 and G2. In G3 the percentage of hepatocytes, sinusoids, and collagen did not differ significantly from the other groups. It was concluded that the ethanolic extract of the red propolis exerted a hepatoprotective effect, because it promoted in vitro reduction of the damage to the DNA of liver cells, antineoplastic activity in human hepatocellular carcinoma cell line (HepG2) and did not exert cytotoxic effect in normal cells or was able to reduce liver enzyme activity and the severity of cirrhosis induced by TAA in vivo.

RESUMO: Este estudo objetivou avaliar o efeito hepatoprotetor do extrato etanólico da própolis vermelha da abelha Apis mellifera, obtido em quatro municípios do semiárido do Rio Grande do Norte, mediante avaliação in vitro do potencial antineoplásico em linhagens de células de carcinoma hepático humano (HepG2) e em linhagens de células normais (L929), além do ensaio cometa em linhagens de células hepáticas (hepatócitos ZF-L) para avaliar o potencial genoprotetor do extrato. O efeito hepatoprotetor também foi avaliado in vivo através da indução de lesões hepática experimental crônica em roedores da espécie Rattus norvegicus (Berkenhout, 1769), linhagem Wistar, pela administração intraperitoneal de tioacetamida (TAA) na dose de 0,2g/kg. Os animais foram distribuídos nos seguintes grupos experimentais: G1 (controle), G2 (tratados com 500mg/kg de extrato etanólico da própolis), G3 (tratados com 500mg/kg de extrato etanólico e TAA) e G4 (tratados com TAA). Todos os ratos foram submetidos aos exames bioquímico sérico, anatomopatológico macroscópico, histológico e esteriológico do fígado. Foi constatado o efeito genoprotetor da própolis vermelha uma vez que as médias dos danos promovidos ao DNA em células testadas com o extrato foram significativamente inferiores à média dos danos do controle positivo (peróxido de hidrogênio). O extrato da própolis vermelha não apresentou atividade citotóxica para células tumorais de câncer de fígado humano, bem como para normais. A ausência de citotoxicidade em células normais, tal como constatado, pode indicar segurança no uso do extrato da própolis. Os resultados da avaliação bioquímica sérica demonstraram que os níveis séricos das enzimas aminotransferase (AST) não diferiram significativamente entre G1, G2 e G3, quando comparadas entre si. No G4 houve aumento significativo dos níveis em relação aos demais grupos, indicando que a administração do extrato não causou toxicidade hepática, bem como exerceu efeito hepatoprotetor frente ao dano hepático induzido pela TAA. Os animais dos G3 e G4 desenvolveram cirrose, porém no G3 os fígados caracterizaram-se pela presença de pequenos nódulos regenerativos e nivelados com a superfície do órgão, enquanto que no G4 os fígados apresentaram grandes nódulos regenerativos. Os fígados dos animais G1 e G2 apresentaram aspecto histológico normal, enquanto que os fígados dos animais do G3 apresentaram nódulos regenerativos circundados por finos septos de tecido conjuntivo, e nos do G4 os nódulos regenerativos foram circundados por espessos septos de tecido conjuntivo fibroso. A análise dos tecidos hepáticos por meio de estereologia mostrou que não houve diferença estatística entre o percentual de hepatócitos, sinusoides e colágenos nos G1 e G2. No G3 o percentual de hepatócitos, sinusoides e colágeno não diferiu significativamente dos demais grupos. Concluiu-se que o extrato etanólico da própolis vermelha exerceu efeito genoprotetor, por promover in vitro redução do dano ao DNA de células hepáticas, atividade antineoplásica em linhagem celular de carcinoma hepatocelular humano (HepG2) e não exerceu efeito citotóxico em células normais ou efeito hepatoprotetor in vivo com diminuição da gravidade da cirrose induzida por TAA.

Hepatoprotective and antineoplastic potencial of red propolis produced by the bees Apis mellifera in the semiarid of Rio Grande do Norte, Brazil / Potencial hepatoprotetor e antineoplásico da própolis vermelha produzida pela abelha Apis mellifera no semiárido do Rio Grande do Norte, Brasil

The objective of this study was to evaluate the hepatoprotective effect of the honey bee Apis mellifera ethanolic extract of the red propolis, obtained in four municipalities of the Rio Grande do Norte semi-arid region, through an in vitro evaluation of the antineoplastic potential in human hepatic carcinoma (HepG2) and normal cell lines (L929), and from the comet assay in hepatic cell lines (ZF-L hepatocytes) to evaluate the genoprotective potential of the extract. The hepatoprotective effect was also evaluated in vivo by the induction of chronic experimental hepatic lesions in rodents (Rattus norvegicus Berkenhout, 1769), Wistar line, by intraperitoneal administration of thioacetamide (TAA) at the dose of 0.2g/kg. The animals were distributed in the following experimental groups: G1 (control), G2 (treated with 500mg/kg ethanolic extract of propolis), G3 (treated with 500mg/kg of ethanolic extract and TAA) and G4 (treated with TAA). All rats were submitted to serum biochemical, macroscopic, histological and stereological biochemical exams of the liver. It was verified the genoprotective effect of red propolis since the mean damages promoted to DNA in cells tested with the extract were significantly lower than the mean of the positive control damage (hydrogen peroxide). The red propolis extract did not present cytotoxic activity to the tumor cells of human liver cancer, as well as to normal ones. The absence of cytotoxicity in normal cells may indicate safety in the use of the propolis extract. The results of the serum biochemical evaluation showed that the serum levels of the aminotransferase enzymes (AST) did not differ significantly between G1, G2 and G3 when compared to each other. G4 showed significant increase in levels compared to the other groups, indicating that the administration of the extract did not cause liver toxicity, as well as exerted hepatoprotective effect against the hepatic damage induced by TAA. The G3 and G4 animals developed cirrhosis, but in G3 the livers were characterized by the presence of small regenerative nodules and level with the surface of the organ, whereas in G4 the livers showed large regenerative nodules. The livers of the G1 and G2 animals presented normal histological appearance, whereas the livers of the G3 animals showed regenerative nodules surrounded by thin septa of connective tissue, and in G4 the regenerative nodules were surrounded by thick septa fibrous connective tissue. The analysis of the hepatic tissues by means of stereology showed that there was no statistical difference between the percentage of hepatocytes, sinusoids, and collagens in G1 and G2. In G3 the percentage of hepatocytes, sinusoids, and collagen did not differ significantly from the other groups. It was concluded that the ethanolic extract of the red propolis exerted a hepatoprotective effect, because it promoted in vitro reduction of the damage to the DNA of liver cells, antineoplastic activity in human hepatocellular carcinoma cell line (HepG2) and did not exert cytotoxic effect in normal cells or was able to reduce liver enzyme activity and the severity of cirrhosis induced by TAA in vivo.(AU)

Este estudo objetivou avaliar o efeito hepatoprotetor do extrato etanólico da própolis vermelha da abelha Apis mellifera, obtido em quatro municípios do semiárido do Rio Grande do Norte, mediante avaliação in vitro do potencial antineoplásico em linhagens de células de carcinoma hepático humano (HepG2) e em linhagens de células normais (L929), além do ensaio cometa em linhagens de células hepáticas (hepatócitos ZF-L) para avaliar o potencial genoprotetor do extrato. O efeito hepatoprotetor também foi avaliado in vivo através da indução de lesões hepática experimental crônica em roedores da espécie Rattus norvegicus (Berkenhout, 1769), linhagem Wistar, pela administração intraperitoneal de tioacetamida (TAA) na dose de 0,2g/kg. Os animais foram distribuídos nos seguintes grupos experimentais: G1 (controle), G2 (tratados com 500mg/kg de extrato etanólico da própolis), G3 (tratados com 500mg/kg de extrato etanólico e TAA) e G4 (tratados com TAA). Todos os ratos foram submetidos aos exames bioquímico sérico, anatomopatológico macroscópico, histológico e esteriológico do fígado. Foi constatado o efeito genoprotetor da própolis vermelha uma vez que as médias dos danos promovidos ao DNA em células testadas com o extrato foram significativamente inferiores à média dos danos do controle positivo (peróxido de hidrogênio). O extrato da própolis vermelha não apresentou atividade citotóxica para células tumorais de câncer de fígado humano, bem como para normais. A ausência de citotoxicidade em células normais, tal como constatado, pode indicar segurança no uso do extrato da própolis. Os resultados da avaliação bioquímica sérica demonstraram que os níveis séricos das enzimas aminotransferase (AST) não diferiram significativamente entre G1, G2 e G3, quando comparadas entre si. No G4 houve aumento significativo dos níveis em relação aos demais grupos, indicando que a administração do extrato não causou toxicidade hepática, bem como exerceu efeito hepatoprotetor frente ao dano hepático induzido pela TAA. Os animais dos G3 e G4 desenvolveram cirrose, porém no G3 os fígados caracterizaram-se pela presença de pequenos nódulos regenerativos e nivelados com a superfície do órgão, enquanto que no G4 os fígados apresentaram grandes nódulos regenerativos. Os fígados dos animais G1 e G2 apresentaram aspecto histológico normal, enquanto que os fígados dos animais do G3 apresentaram nódulos regenerativos circundados por finos septos de tecido conjuntivo, e nos do G4 os nódulos regenerativos foram circundados por espessos septos de tecido conjuntivo fibroso. A análise dos tecidos hepáticos por meio de estereologia mostrou que não houve diferença estatística entre o percentual de hepatócitos, sinusoides e colágenos nos G1 e G2. No G3 o percentual de hepatócitos, sinusoides e colágeno não diferiu significativamente dos demais grupos. Concluiu-se que o extrato etanólico da própolis vermelha exerceu efeito genoprotetor, por promover in vitro redução do dano ao DNA de células hepáticas, atividade antineoplásica em linhagem celular de carcinoma hepatocelular humano (HepG2) e não exerceu efeito citotóxico em células normais ou efeito hepatoprotetor in vivo com diminuição da gravidade da cirrose induzida por TAA.(AU)

Carbonyl compounds in wine: factors related to presence and toxic effects / Compostos carbonílicos em vinho: fatores relacionados a presença e efeitos tóxicos

The objective of this study was to review technological and toxicological factors related to presence of carbonyl compounds found in wines, including acetaldehyde, formaldehyde, acrolein, ethyl carbamate (EC) and furfural. Acetaldehyde and formaldehyde may be formed through the ethanol and methanol oxidation, respectively. Acrolein may arise as a thermal degradation product of glycerol, amino acids, carbohydrates and triglycerides or by metabolic activity of microorganisms. In addition, acrolein and furfural are formed during wood combustion; therefore, these aldehydes may be present in raw materials due to the environmental contamination. Furfural is also a product of the Maillard reaction formed from sugars and amino acids, while ethyl carbamate occurs through the reaction between urea and ethanol. These compounds may react with SO2 and phenolic compounds to form non-volatile adducts, which positively modulates color stability, astringency and aroma in wine. However, when ingested through wine, electrophilic carbonyl compounds may form adducts with nucleophilic targets, such as DNA, resulting in genotoxicity along the gastrointestinal tract. Furthermore, carbonyl compounds induce the increase of reactive oxygen species and can trigger apoptosis, in addition to hepatocellular adenoma and carcinoma as a consequence of chronic hepatotoxicity. Neurodegenerative diseases may be related to the exposure to carbonyl compounds. Therefore, strategies to reduce the levels of these compounds should be studied in order to get the most out of the beneficial functional properties of wine consumption.(AU)

O objetivo deste estudo foi revisar os fatores tecnológicos e toxicológicos relacionados à presença de compostos carbonílicos encontrados em vinhos, incluindo acetaldeído, formaldeído, acroleína, carbamato de etila (CE) e furfural. O acetaldeído e o formaldeído podem ser formados através da oxidação do etanol e do metanol, respectivamente. A acroleína pode surgir como um produto de degradação térmica de glicerol, aminoácidos, carboidratos e triglicerídeos ou pela atividade metabólica de microorganismos. Além disso, a acroleína e o furfural são formados durante a combustão da madeira. Portanto, esses aldeídos podem estar presentes nas matérias-primas devido à contaminação ambiental. O furfural é também um produto da reação de Maillard formado a partir de açúcares e aminoácidos, enquanto o carbamato de etila ocorre através da reação entre uréia e etanol. Estes compostos podem reagir com SO2 e compostos fenólicos para formar adutos não voláteis, que modulam positivamente a estabilidade da cor, adstringência e aroma no vinho. No entanto, quando ingeridos através do vinho, os compostos carbonílicos que são eletrofílicos podem formar adutos com alvos nucleofílicos, como o DNA, resultando em genotoxicidade ao longo do trato gastrintestinal. Além disso, os compostos carbonílicos também induzem o aumento de espécies reativas de oxigênio e podem desencadear a apoptose, além de adenoma e carcinoma hepatocelular como consequência da hepatotoxicidade crônica. Doenças neurodegenerativas podem estar relacionadas à exposição aos compostos carbonílicos. Com isso, estratégias para reduzir os níveis desses compostos devem ser estudadas para obter o máximo das propriedades funcionais benéficas do consumo de vinho.(AU)

In vivo induction of hepatocellular carcinoma by diethylnitrosoamine and pharmacological intervention in Balb C mice using Bergenia ciliata extracts

Abstract Background Hepatocellular carcinoma is the most frequent primary malignancy of liver and accounts for as many as one million deaths worldwide in a year. Objectives The aim of the present study was to evaluate the anti-cancerous efficiency of Bergenia ciliata rhizome against diethylnitrosoamine induced hepatocarcinogenesis in Balb C mice. Methods One percent diethylnitrosoamine was prepared by using 99 ml of normal saline NaCl (0.9 percent) solution to which was added 1 ml of concentrated diethylnitrosoamine (DEN) solution (0.01 g/l). Extract of Bergenia ciliata was prepared by maceration technique. Mice were classified into four groups as follows: Group 1 a control group (N=7) received saline solution (3.5 l/mg), group 2 (N=14) received diethylnitrosoamine (3.5 l/mg) intraperitoneally once in a week for eight consecutive weeks, group 3 (N=7) received plant extract (150 mg/kg (Body weight)) once in a week, while group 4 (N=7) was given combination of diethylnitrosoamine (3.5 l/mg) and plant extract (150 mg/kg (Body weight)). After eight weeks of DEN induction group 2 mice were divided into two subgroups containing seven mice each, subgroup 1 was sacrificed while subgroup 2 was treated with plant extract (150 mg/kg (Body weight)) once in a week for eight consecutive weeks. Results The model of DEN injected hepatocellular carcinomic (HCC) mice elicited significant decline in levels of albumin with concomitant significant elevations in tumor markers aspartate aminotransferase, alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alpha feto protein (AFP), gamma glutamyl transferase (Y-GT), 5 nucleotidase (5NT), glucose-6-phosphate dehydrogenase (G6PDH) and bilirubin. The intraperitoneal administration of B. ciliata as a protective agent, produced significant increase in albumin levels with significant decrease in the levels of tumor markers aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alpha feto protein (AFP), gamma glutamyl transferase (Y-GT), 5 nucleotidase (5NT), glucose-6-phosphate dehydrogenase (G6PDH) and bilirubin. Conclusion Bergenia ciliata has potent antioxidant activity, radical scavenging capacity and anticancerous properties. Bergenia ciliata extracts may provide a basis for development of anti-cancerous drug.

Resumo Antecedentes O carcinoma hepatocelular é a neoplasia primária mais frequente do fígado e é responsável por até um milhão de mortes em todo o mundo em um ano. Objetivos O objetivo do presente estudo foi avaliar a eficiência anticancerígena do rizoma de Bergenia ciliata contra a hepatocarcinogênese induzida por dietilnitrosoamina em camundongos balb c. Métodos Um por cento de dietilnitrosoamina foi preparado usando 99 ml de solução salina normal (0,9 por cento) à qual foi adicionado 1 ml de solução concentrada de dietilnitrosoamina (DEN) (0,01 g / l). O extrato de Bergenia ciliata foi preparado pela técnica de maceração. Os ratos foram classificados em quatro grupos: Grupo 1 grupo controle (N = 7) recebeu solução salina (3,5 mL / mg), grupo 2 (N = 14) recebeu dietilnitrosoamina (3,5 mL / mg) por via intraperitoneal uma vez por semana para oito semanas consecutivas, o grupo 3 (N = 7) recebeu extrato vegetal (150 mg / kg (peso corporal)) uma vez por semana, enquanto o grupo 4 (N = 7) recebeu combinação de dietilnitrosoamina (3,5 l / mg) e extrato (150 mg / kg (peso corporal). Após oito semanas do grupo de indução DEN 2 ratos foram divididos em dois subgrupos contendo sete ratos cada, subgrupo 1 foi sacrificado enquanto subgrupo 2 foi tratado com extrato vegetal (150 mg / kg)) uma vez por semana durante oito semanas consecutivas. Resultados O modelo de camundongos hepatocelulares carcinômicos (CHC) injetados com DEN provocou declínio significativo nos níveis de albumina com elevações significativas concomitantes nos marcadores tumorais: aspartato aminotransferase, alanina aminotransferase (ALT), lactato desidrogenase (LDH), proteína alfa feto (AFP), gama glutamiltransferase (Y-GT), 5 nucleotidase (5NT), glicose-6-fosfato ehidrogenase (G6PDH) e bilirrubina. A administração intraperitoneal de B. ciliata como agente protetor produziu um aumento significativo nos níveis de albumina com uma diminuição significativa nos níveis dos marcadores tumorais: aspartato aminotransferase, alanina aminotransferase (ALT), lactato desidrogenase (LDH), proteína alfa feto (AFP), gama glutamiltransferase (Y-GT), 5 nucleotidase (5NT), glicose-6-fosfato desidrogenase (G6PDH) e bilirrubina. Conclusão Bergenia ciliata possui atividade antioxidante potente, capacidade de eliminação de radicais livres e propriedades anticancerígenas. Extratos de Bergenia ciliata podem fornecer uma base para o desenvolvimento de drogas anti-cancerígenas.