Expression of LIN28A/B and Let-7 miRNAs in canine mammary carcinomas / Expressão de LIN28A/B e Let-7 miRNAs em carcinomas mamários caninos

LIN28 is a RNA-binding protein including two highly conserved homologous, LIN28A and LIN28B. Proto-oncogenes such as LIN28A and LIN28B are generally targeted by the let-7 miRNAs in different types of human cancers. Here, we determined the expression of LIN28A in canine mammary tumor samples and the LIN28/let-7 pathway in canine mammary cell lines. In those cell lines, we identified a functional LIN28/let-7 pathway which exhibited high expression of let-7 members and low expression of its targets, including LIN28A and LIN28B. However, the mammary carcinoma tissue samples showed a frequent expression of LIN28A being expressed mainly in the epithelial cells. No association was observed between LIN28A expression and histopathological classification and grade, TNM and survival time. Our results suggested a possible role of the LIN28A protein in the development of canine mammary carcinomas due to the high frequency observed in the tumor samples (28 of 32). The in vitro experiments suggested that the LIN28/let-7 pathway is active in the tumor cells evaluated. However, more studies are necessary to elucidate the exact role of LIN28/let-7 pathway in canine mammary carcinomas.

LIN28 é uma proteína de ligação ao RNA, com duas formas homólogas altamente conservadas, LIN28A e LIN28B. Os proto-oncogenes LIN28A e LIN28B são regulados pela família de miRNAs let-7 em diferentes tipos de cânceres em humanos. No presente trabalho, o objetivo foi determinar a expressão de LIN28A em amostras de tumor mamário de cadelas e a via LIN28/let-7 em linhagens celulares mamárias caninas. Nestas linhagens, através das técnicas de qPCR e RNAseq, foi identificado que a via LIN28/let-7 apresenta-se funcional, com alta expressão dos membros da família let-7 e baixa expressão de seus alvos, entre eles LIN28A e LIN28B. No entanto, as amostras de tecidos de carcinomas mamários caninos demonstraram expressão frequente de LIN28A, sendo observada principalmente em células epiteliais. Não foram observadas associações entre expressão de LIN28A com classificação e gradação histopatológicas, TNM e tempo de sobrevida. Nossos resultados sugerem uma possível relação da proteína LIN28A no desenvolvimento de carcinomas mamários caninos devido à alta frequência observada nas amostras tumorais (28 de 32). Os experimentos in vitro sugerem que a via LIN28/let-7 é ativa nas linhagens celulares caninas avaliadas. Entretanto, estudos funcionais ainda são necessários para elucidar a função exata da via LIN28/let-7 nos carcinomas mamários caninos.

Acute phase protein and vitamin D concentration in dogs with multicentric lymphoma / Concentração de proteínas de fase aguda e vitamina D em cães com linfoma multicêntrico

This study aimed to evaluate the serum concentration of vitamin D (25-Hydroxyvitamin D) and acute phase proteins (APPs; alpha-1 acid glycoprotein, haptoglobin, transferrin, ceruloplasmin, albumin, IgA, IgG and alpha-1 - antitrypsin) as potential biomarkers for prognostic and therapy response in dogs with multicentric lymphoma submitted to the CHOP (Cyclophosphamide, Doxorubicin, Vincristine and Prednisone) chemotherapy protocol. Thirteen dogs with multicentric lymphoma classified as high grade by cytology were included in the treatment group (GL), while ten healthy dogs were included in the control group (GC). Serum was collected in the weeks T0, T5 and T10 of CHOP chemotherapy protocol, for the GL group, and in a single collection, for the GC group. All the collected samples were evaluated for the APPs and vitamin D concentrations through electrophoresis and chemiluminescence methods, respectively. Diagnostic and staging tests were performed for all the dogs in the GL group, and included cytopathology, histopathology and immunohistochemistry of the affected lymph node. Of these dogs, 9 achieved a complete response and 4 a partial response to the treatment. Data analysis was performed with the R software. The results demonstrated that serum concentrations of IgA, haptoglobin and α1-acid glycoprotein were significantly different between the groups and also between the different chemotherapy times analyzed (p<0.05), indicating that these proteins can be considered as sensitive biomarkers for lymphoma in dogs. Furthermore, the α1-acid glycoprotein showed prognostic value for the disease, with 63% specificity. However, vitamin D concentration was not correlated with prognosis of the dogs with lymphoma.

Objetivou-se caracterizar a concentração sérica da vitamina D e das PFAs (Proteínas de Fase Aguda) (alfa-1 glicoproteína ácida, haptoglobina, transferrina, ceruloplasmina, albumina, IgA, IgG e alfa-1 ­ antitripsina) em cães com linfoma multicêntrico, submetidos ao tratamento quimioterápico com protocolo CHOP (Ciclofosfamida, Doxorrubicina, Vincristina e Prednisona), determinando o valor prognóstico desses marcadores para a doença. Foram avaliadas as concentrações séricas das PFAs, através do método da eletroforese e as concentrações da vitamina D, através da quimioluminescência em dois grupos experimentais, um grupo de 13 cães com linfoma multicêntrico classificados como alto grau pela citologia (GL) durante as semanas T0, T5 e T10 do tratamento com protocolo quimioterápico antineoplásico e em um grupo de 10 animais saudáveis para compor o grupo controle (GC), em coleta única. Para isso, foi realizado o diagnóstico, estadiamento e avaliação de resposta terapêutica dos 13 pacientes com linfoma multicêntrico através de técnicas de citopatologia, histopatologia, imuno-histoquímica do linfonodo periférico acometido. Foi observado que 9 pacientes tiveram resposta completa e 4 pacientes tiveram resposta parcial ao tratamento. Os dados foram analisados através do software R. Os resultados indicam que as diferenças entre as variáveis IgA, haptoglobina e α1-glicoproteína ácida foram significativas entre os grupos, e entre os diferentes momentos da quimioterapia (p< 0,05), indicando que podem ser marcadores sensíveis ao linfoma em cães. A α1-glicoproteína ácida apresentou valor prognóstico para o linfoma, com 63% de especificidade. Porém a vitamina D não apresentou valor prognóstico para o linfoma multicêntrico em cães.

Matrix metalloproteinase 9 expression in canine mammary carcinomas

Mammary neoplasms are among the most common canine tumors, with high risk of invasion and metastasis. Important steps for these events are the loss of cell adhesion to the main tumor mass and extracellular matrix degradation. Matrix metalloproteinases (MMPs) are proteolytic enzymes containing zinc, which are capable of degrading and remodeling the surrounding extracellular matrix, facilitating these events. Involvement of MMPs has been demonstrated in many pathological processes, as well as in human and canine tumors, which has been related to malignancy and prognosis. The aim of this study was to characterize the immunohistochemical expression of matrix metalloproteinase 9 (MMP-9) in canine mammary gland tumors, as well as to verify its relation with the different histologic patterns. Thirty-one of the 41 tumors (75.61%) were positive. Twenty-two samples (70.97%) had diffuse cytoplasmic immunostaining and 9 (29.03%) had a finely granular pattern. Immunostaining intensity was strong in 21 (67.74%) tumors and weak in 10 (32.26%). No statistically significant differences were found between anaplastic carcinomas, carcinomas in a mixed tumor and simple carcinomas regarding positivity (p=0.9707), intensity (p=0.5386) and staining pattern (0.6135); between solid carcinomas and simple papillary carcinomas for positivity (p=0,7333), intensity (p=0.7333) and staining pattern (p=0.3037); or betweensolid carcinomas and simple tubular and papillary carcinomas for positivity (p=0.9682), intensity (p=0.8450) and staining pattern (p=0.5068). MMP-9 was detected with variable intensity and morphological patterns of cytoplasmic staining. However significant statistic differences were not found between the histological types or histopathological grades.

Matrix metalloproteinase 9 expression in canine mammary carcinomas

Mammary neoplasms are among the most common canine tumors, with high risk of invasion and metastasis. Important steps for these events are the loss of cell adhesion to the main tumor mass and extracellular matrix degradation. Matrix metalloproteinases (MMPs) are proteolytic enzymes containing zinc, which are capable of degrading and remodeling the surrounding extracellular matrix, facilitating these events. Involvement of MMPs has been demonstrated in many pathological processes, as well as in human and canine tumors, which has been related to malignancy and prognosis. The aim of this study was to characterize the immunohistochemical expression of matrix metalloproteinase 9 (MMP-9) in canine mammary gland tumors, as well as to verify its relation with the different histologic patterns. Thirty-one of the 41 tumors (75.61%) were positive. Twenty-two samples (70.97%) had diffuse cytoplasmic immunostaining and 9 (29.03%) had a finely granular pattern. Immunostaining intensity was strong in 21 (67.74%) tumors and weak in 10 (32.26%). No statistically significant differences were found between anaplastic carcinomas, carcinomas in a mixed tumor and simple carcinomas regarding positivity (p=0.9707), intensity (p=0.5386) and staining pattern (0.6135); between solid carcinomas and simple papillary carcinomas for positivity (p=0,7333), intensity (p=0.7333) and staining pattern (p=0.3037); or betweensolid carcinomas and simple tubular and papillary carcinomas for positivity (p=0.9682), intensity (p=0.8450) and staining pattern (p=0.5068). MMP-9 was detected with variable intensity and morphological patterns of cytoplasmic staining. However significant statistic differences were not found between the histological types or histopathological grades.(AU)

Chemical carcinogenesis by DMBA (7, 12-dimethylbenzanthracene) in female BALB/c mice: new facts / Carcinogenese quimica por DMBA (7, 12-dimethylbenzanthracene) em camundongos femeas BALB/c: novos fatos

Polycyclic aromatic hydrocarbons are known carcinogens used in rodent experimental models. In this study, the carcinogen DMBA (7,12-dimethylbenzanthracene) was administered by gavage, diluted in corn oil, to female BALB / c mice at hebdomadary doses of 1 mg per animal for 1, 3, 6 or 9 weeks. Animals were weighed and monitored weekly until death. Remaining animals were euthanized at the age of 53 weeks. At necropsy, representative fragments of neoplasms were collected and routinely processed for histopathological analysis. Of all mice that received DMBA, 68.57% developed some type of tumor. Of the 70 mice treated with various doses of DMBA, 22 (31.43%) developed mammary tumors. The adenoacanthoma was the most commonly (18.75%) diagnosed histological type of breast cancer. Lung (15.71%), lymphoid tissue (11.43%), stomach (7.14%) and skin (2.86%) were also primary sites of tumor development. One third (33.33%) of the mice receiving 1 mg of DMBA developed lung cancer. Therefore, the administration of DMBA was shown to be an efficient model of carcinogenesis in mice, especially for the study of breast cancer, when using the highest dose, and lung, when using the lowest dose. Carcinogenesis models have been used for several purposes in cancer research. These results represent new facts for a classic carcinogenesis model. (AU)

Hidrocarbonetos policíclicos e aromáticos são carcinógenos usados em modelos experimentais em roedores. Neste estudo, o carcinógeno DMBA (7,12-dimetilbenzantraceno) foi administrado por gavagem, diluído em óleo de milho, para camundongos BALB/c em doses hebdomadárias de 1 mg por animal por 1, 3, 6 ou 9 semanas. Os animais foram pesados e monitorados semanalmente até a morte. Os animais remanescentes foram eutanasiados com a idade de 53 semanas. Na necroscopia, fragmentos representativos das neoplasias foram colhidos e rotineiramente processados para exame histopatológico. De todos os animais que receberam DMBA, 68,57% desenvolveram algum tipo de tumor. Entre os 70 camundongos tratados com diferentes doses de DMBA, 22 (31,43%) desenvolveram neoplasias mamárias. O adenoacantoma foi o tumor mamário mais comumente diagnosticado (18,75%). Pulmões (15,71%), tecido linfoide (11,43%), estômago (7,14%) e pele (2,86%) foram também locais primários de desenvolvimento de neoplasias. Um terço (33,33%) dos camundongos que receberam 1 mg de DMBA desenvolveram neoplasias pulmonares. Portanto, a administração de DMBA foi considerada um modelo eficiente de carcinogênese em camundongos, especialmente para o estudo de neoplasias mamárias, quando a maior dose é utilizada, e de neoplasias pulmonares, quando utilizada a menor dose. Os modelos de carcinogênese química têm sido usados para diversos estudos na pesquisa em câncer, os resultados aqui apresentados mostram novos fatos para um modelo clássico de carcinogênese. (AU)

Selenium and vitamin E enriched diet increases NK cell cytotoxicity in cattle / Dieta enriquecida com selênio e vitamina E aumenta a citotoxicidade de células NK em bovinos

A number of studies has shown that antioxidants, fatty acids and trace minerals may modulate different immune cell activities, and that their deficiency may be associated with diseases and impaired immune responses. In innate immunity, natural killer (NK) cells have a central role, killing virally infected and cancerous cells, and also secreting cytokines that shape adaptive immune responses. Thus, the aim of this study was to evaluate the effect of enriched diets in selenium plus vitamin E and/or canola oil on complete blood count and on NK cell cytotoxicity from blood lymphocytes of Nellore bulls. Bulls that received selenium plus vitamin E had (P=0.0091) higher NK cell cytotoxicity than control bulls. This result positively correlated with serum selenium levels. To the best of our knowledge, this is the first study that showed immunostimulatory effects of selenium plus vitamin E on NK cell cytotoxicity of Nellore bulls.(AU)

Vários estudos demonstraram que antioxidantes, ácidos graxos e minerais podem modular a atividade de diferentes células do sistema imunológico e que as suas carências podem estar associadas a doenças e a respostas imunes comprometidas. Na imunidade inata, os linfócitos natural killer (NK) têm um papel central matando células infectadas por vírus e células cancerígenas, ao mesmo tempo em que também secretam citocinas que modulam as respostas imunes adaptativas. Assim, o objetivo deste estudo foi avaliar o efeito de dietas enriquecidas em selênio e vitamina E e/ou óleo de canola no hemograma e na citotoxicidade das células NK do sangue de bovinos da raça Nelore. Os animais que receberam selênio e vitamina E tiveram (P = 0,0091) maior citotoxicidade das células NK do que os animais do grupo controle. Este resultado foi positivamente correlacionado com os níveis de selênio no sangue. Para o melhor do nosso conhecimento, este é o primeiro estudo que mostrou efeitos imunoestimulatórios do selênio e vitamina E sobre a citotoxicidade das células NK de bovinos Nelore.(AU)

Selenium and vitamin E enriched diet increases NK cell cytotoxicity in cattle / Dieta enriquecida com selênio e vitamina E aumenta a citotoxicidade de células NK em bovinos

A number of studies has shown that antioxidants, fatty acids and trace minerals may modulate different immune cell activities, and that their deficiency may be associated with diseases and impaired immune responses. In innate immunity, natural killer (NK) cells have a central role, killing virally infected and cancerous cells, and also secreting cytokines that shape adaptive immune responses. Thus, the aim of this study was to evaluate the effect of enriched diets in selenium plus vitamin E and/or canola oil on complete blood count and on NK cell cytotoxicity from blood lymphocytes of Nellore bulls. Bulls that received selenium plus vitamin E had (P=0.0091) higher NK cell cytotoxicity than control bulls. This result positively correlated with serum selenium levels. To the best of our knowledge, this is the first study that showed immunostimulatory effects of selenium plus vitamin E on NK cell cytotoxicity of Nellore bulls.(AU)

Vários estudos demonstraram que antioxidantes, ácidos graxos e minerais podem modular a atividade de diferentes células do sistema imunológico e que as suas carências podem estar associadas a doenças e a respostas imunes comprometidas. Na imunidade inata, os linfócitos natural killer (NK) têm um papel central matando células infectadas por vírus e células cancerígenas, ao mesmo tempo em que também secretam citocinas que modulam as respostas imunes adaptativas. Assim, o objetivo deste estudo foi avaliar o efeito de dietas enriquecidas em selênio e vitamina E e/ou óleo de canola no hemograma e na citotoxicidade das células NK do sangue de bovinos da raça Nelore. Os animais que receberam selênio e vitamina E tiveram (P = 0,0091) maior citotoxicidade das células NK do que os animais do grupo controle. Este resultado foi positivamente correlacionado com os níveis de selênio no sangue. Para o melhor do nosso conhecimento, este é o primeiro estudo que mostrou efeitos imunoestimulatórios do selênio e vitamina E sobre a citotoxicidade das células NK de bovinos Nelore.(AU)

Consensus for the Diagnosis, Prognosis and Treatment of Canine Mammary Tumors 2013

The purpose of this paper is to establish criteria that could guide the diagnosis, prognosis and treatment of caninemammary neoplasias. It was elaborated during the Mammary Pathology Meeting: Diagnosis, Prognosis and Treatment ofthe Canine Mammary Neoplasia, held on November 6th and 7th, 2010 in Belo Horizonte – MG, Brazil. Academics fromseveral regions of Brazil were present and contributed to this work. After three years, a new discussion was foundnecessary in order to address important questions: 1 - Have Brazilian DVMs applied the consensus? 2 - What were the maindifficulties in applying the consensus? 3 - What were the obtained results? 4 - What were the main differences among thevarious oncology services/groups? 5 - How could the criteria be improved and uniformed? A spreadsheet that allowed datacollection and an abstract was submitted by each oncology service/group from various parts of the country. Based on theabstracts we identified the main differences in diagnosis and therapeutic conducts among the groups. These differences haveguided the discussions of the II Mammary Pathology Meeting and the publication of a second consensus that has beenrevised and updated. The II Mammary Pathology Meeting: Diagnosis, Prognosis and Treatment of the Canine mamaryNeoplasia, was held on December 9th, 10thand 11th, 2013 in Belo Horizonte – MG, sponsored by the Laboratory ofComparative Pathology of the Federal University of Minas Gerais (UFMG), with the support of the Brazilian Associationof Veterinary Pathology (ABPV) and Brazilian Association of Veterinary Oncology (ABROVET). Academics from severalregions of Brazil were present and contributed to this work.

Consensus for the Diagnosis, Prognosis and Treatment of Canine Mammary Tumors 2013

The purpose of this paper is to establish criteria that could guide the diagnosis, prognosis and treatment of caninemammary neoplasias. It was elaborated during the Mammary Pathology Meeting: Diagnosis, Prognosis and Treatment ofthe Canine Mammary Neoplasia, held on November 6th and 7th, 2010 in Belo Horizonte MG, Brazil. Academics fromseveral regions of Brazil were present and contributed to this work. After three years, a new discussion was foundnecessary in order to address important questions: 1 - Have Brazilian DVMs applied the consensus? 2 - What were the maindifficulties in applying the consensus? 3 - What were the obtained results? 4 - What were the main differences among thevarious oncology services/groups? 5 - How could the criteria be improved and uniformed? A spreadsheet that allowed datacollection and an abstract was submitted by each oncology service/group from various parts of the country. Based on theabstracts we identified the main differences in diagnosis and therapeutic conducts among the groups. These differences haveguided the discussions of the II Mammary Pathology Meeting and the publication of a second consensus that has beenrevised and updated. The II Mammary Pathology Meeting: Diagnosis, Prognosis and Treatment of the Canine mamaryNeoplasia, was held on December 9th, 10thand 11th, 2013 in Belo Horizonte MG, sponsored by the Laboratory ofComparative Pathology of the Federal University of Minas Gerais (UFMG), with the support of the Brazilian Associationof Veterinary Pathology (ABPV) and Brazilian Association of Veterinary Oncology (ABROVET). Academics from severalregions of Brazil were present and contributed to this work.(AU)

Consensus for the Diagnosis, Prognosis and Treatment of Canine Mammary Tumors

The purpose of this paper is to establish criteria that could guide the diagnosis, prognosis and treatment of canine mammary neoplasias. It was elaborated during the Mammary Pathology Meeting: Diagnosis, Prognosis and Treatment of the Canine Mammary Neoplasm, held on November 6th and 7th, 2010 in Belo Horizonte ­ MG, Brazil, sponsored by the Laboratory of Comparative Pathology ­ UFMG, with the support of the Brazilian Association of Veterinary Pathology (ABPV) and Brazilian Association of Veterinary Oncology (ABROVET). Academics from several regions of Brazil were present and contributed to this work.(AU)

Diminished angiogenesis in the cornea of mice with heterologous deletion of Connexin 43 gene (Gja1)

Angiogenesis is involved in several physiological and pathological processes, and the proliferation of endothelial cells is a central event in the generation of new blood vessels. Gap junctions (GJ) are membrane structures that communicate adjacent cells, contribute to tissue homeostasis, and are important to the control of cell proliferation. Connexins (Cxs) are the proteins that form gap junctions. Endothelial cells may express Cx43, Cx37 and Cx40. In this study, we analyzed the effect of the heterologous deletion of the Gja1 (Cx43 gene) on the development of newly formed blood vessels (NFBV) in the mouse cornea. Heterozygous (Cx43+/-) and wild-type (Cx43+/+) mice were submitted to the silver crystal corneal cauterization model. Two parameters were quantified by image analysis 2 or 6 days after cauterization: NFBV density and length. At days 2 and 6 after corneal cauterization, Cx43+/- mice showed smaller density of NFBV as compared to Cx43+/+ mice. Therefore, deletion of one Gja1 allele (connexin-43 gene) may lead to impaired cell-cell communication in endothelial cells, diminishing angiogenesis after cauterization of the mouse cornea

Diminished angiogenesis in the cornea of mice with heterologous deletion of Connexin 43 gene (Gja1)

Angiogenesis is involved in several physiological and pathological processes, and the proliferation of endothelial cells is a central event in the generation of new blood vessels. Gap junctions (GJ) are membrane structures that communicate adjacent cells, contribute to tissue homeostasis, and are important to the control of cell proliferation. Connexins (Cxs) are the proteins that form gap junctions. Endothelial cells may express Cx43, Cx37 and Cx40. In this study, we analyzed the effect of the heterologous deletion of the Gja1 (Cx43 gene) on the development of newly formed blood vessels (NFBV) in the mouse cornea. Heterozygous (Cx43+/-) and wild-type (Cx43+/+) mice were submitted to the silver crystal corneal cauterization model. Two parameters were quantified by image analysis 2 or 6 days after cauterization: NFBV density and length. At days 2 and 6 after corneal cauterization, Cx43+/- mice showed smaller density of NFBV as compared to Cx43+/+ mice. Therefore, deletion of one Gja1 allele (connexin-43 gene) may lead to impaired cell-cell communication in endothelial cells, diminishing angiogenesis after cauterization of the mouse cornea(AU)

Compensatory kidney hypertrophy/hyperplasia after nephrectomy in mice: alterations of connexin 43 (Cx43) phosphorylated isoforms

Compensatory kidney hypertrophy/hyperplasia leads to several changes in kidney structure and function, as increased glomeruli filtration. The aim of this study was to evaluate connexin 43 in remnant mouse kidneys after unilateral nephrectomy. The right kidney was surgically removed from BALB/c mice. Groups were euthanized at 24, 48 and 72 hours, and at 7 and 30 days. Kidney sections of the reminiscent kidneys were stained with Periodic Acid/Schiff and additional slides were submitted to BrdU and Cx43 immunohistochemistry. The results demonstrated an increase in kidney weight as early as 24 hours through 30 post-nephrectomy. In addition, BrdU positive epithelial cells increased at 24 and 48 hours post-nephrectomy. Cx43 was detected in the cytoplasm and membrane of epithelial cells and vasculature. Taking into consideration the quantity, intensity and localization of Cx43 immunostaining pattern, we observed that nephrectomized mice presented lower Cx43 expression and a cytoplasmic localization after 24 hours, peaking in 48 hours. Furthermore, western blot revealed that during the first 24 and 48 hours after nephrectomy, PO (unphosphorylated) and P1 (phosphorylated) Cx43 disappeared, and the products of Cx43 degradation were reduced. On the other hand, after 72 hours the PO and P1 state reappeared and the amount of degraded peptides also increased. Seven and thirty days after nephrectomy, a higher intensity of PO and P1 state and a lower P2 (hyperphosphorylated) band were observed. In conclusion, our results suggest that Cx43 phosphorylation results in the retention of Cx43 in cytoplasm and its increased degradation during compensatory renal hyperplasia/hypertrophy.

Sobre os efeitos quimiopreventivos e antitumorais do guaraná, Paullinia cupana Mart var. sorbilis, em modelos experimentais in vivo e in vitro / On the chemopreventive and antineoplastic effects of guarana, Paullinia cupana Mart var. sorbilis, in in vivo and in vitro experimental models

O câncer é a segunda maior causa de morte no Brasil, atrás apenas de doenças cardíacas. Por isto, é evidente que grandes recursos sejam direcionados para a pesquisa no descobrimento de novas opções com a finalidade de erradicar esta doença. Dentre estas opções, a quimioprevenção do câncer tem chamado a atenção já que, mesmo com os imensos avanços no conhecimento sobre os mecanismos da carcinogênese e conseqüente desenvolvimento de novas drogas, os dados estatísticos de mortalidade não se tornaram menores. Somando-se a estes fatos, deve ser considerado que no Brasil o tratamento padrão do câncer não chega a todas as pessoas por ser extremamente caro. Desta forma, a quimioprevenção do câncer com fatores presentes na dieta ou oriundos de fontes consideradas baratas como fitoterápicos, deve ser apreciada. Assim, este trabalho teve como objetivo avaliar os efeitos quimiopreventivos e antineoplásicos de uma planta brasileira, o guaraná (Paullinia cupana Mart var. sorbilis). Foram utilizados alguns experimentos em camundongos como indução genotóxica em fígado pela Dietilnitrosamina (DEN); carcinogênese pulmonar induzida pela 4-(metilnitrosamino)-1-(3-piridil)-1-butanona (NNK), uma nitrosamina presente no tabaco; tumor ascítico de Ehrlich; disseminação hematógena de melanoma B16/f10; e cultivo de células tumorais e não tumorais. Além disso, caracterizou-se o papel da Conexin43 na carcinogênese pulmonar induzida pelo NNK e os efeitos do guaraná sobre o receptor CAR e sua ação quando da administração do ligante do CAR, 1,4-bis[2-(3,5-dichloropiridiloxi)]benzeno (TCPOBOP). Pudemos observar efeitos quimiopreventivos e antineoplásicos do guaraná dependendo do modelo utilizado, demonstrando que seu modo de ação principal é a redução da proliferação celular. Além disso, observamos que os tumores de pulmão dos animais tratados com a planta apresentavam menor tamanho, menor grau maligno, menor índice de proliferação celular e menor ativação do fator de transcrição CREB. Observamos também que a Conexina43 (Cx43) tem importante papel na carcinogênese pulmonar induzida pelo NNK, atuando como supressor tumoral e em fases tardias possivelmente tendo papel inverso, ou seja, como um oncogene. Caracterizamos os efeitos do guaraná sobre a ativação do receptor CAR e demonstramos que, por si só, o guaraná induz a expressão do CAR, além de alterar a expressão de alguns de seus transcritos como a CYP2B10 e CYP3A11. Ao analisarmos os efeitos de extratos de guaraná sobre células de tumor de pulmão (E9) in vitro, verificamos o mesmo efeito antiproliferativo, diminuindo a expressão do PCNA e da Conexina43 de maneira dose-dependente, além de verificar um aumento da expressão do receptor CAR. Ao fim propomos uma hipótese de mecanismo de ação baseando-se nas alterações encontradas oriundas da administração do guaraná. Concluímos que o guaraná apresenta componentes com ação antitumoral em camundongos, tendo efeito quimiopreventivo ou antineoplásico dependendo do modelo utilizado

Cancer is the second biggest cause of deaths in Brazil, only behind of cardiac diseases. As a result, it is evident that great resources for research will be directed towards the discovery of new options to eradicate this disease. Among these options, cancer chemoprevention has calling for attention since the huge advances in the knowledge of carcinogenesis and development of new drugs did not decrease statistical data on mortality due to cancer. In addition, it must be considered that in Brazil, cancer therapy is not available for all given that it is too expensive. Therefore, cancer chemoprevention with dietary factors or from medicinal plants has got to be treasured. Following these lines, the aim of this work was to evaluate the chemopreventive and antineoplastic effects of a Brazilian plant, Paullinia cupana Mart var. sorbilis, most known as guarana. It was used several experiments in mice and cell culture essays as: protection against DEN-induced DNA damage; NNK-induced lung carcinogenesis; Ehrlich Ascitic Tumor; metastasis of B16/f10 melanoma cells; and cell culture of a tumorigenic and a non-tumorigenic cell lines. Additionally, it was characterized the role of Connexin43 in the NNK-induced lung carcinogenesis and the effects of guarana on the CAR receptor before and after the administration of TCPOBOP. We note a chemopreventive or antineoplastic effect of guarana depending on the model employed and showed that the mode of action responsible for these effects was reduced cell proliferation. Also, the lung tumors of guarana-treated animals were smaller, less aggressive, with decreased cell proliferation and CREB activation. On the other hand, we observed that Connexin43 have an important role on NNK-induced lung carcinogenesis because it may act as a tumor suppressor and in advanced stages as an oncogene. The effects of guarana on the CAR activation were characterized and we showed that guarana induces CAR mRNA expression, altering the levels of its transcripts as CYP2B10 and CYP3A11. We also examined the effects of guarana extracts on a lung tumor cell culture (E9 cells) and demonstrated the same antiproliferative effect observed previously, by decreased PCNA and Connexin43 proteins in a dose-dependent manner along with an increase in CAR protein. At last we hypothesized a mechanism of action for guarana effects basing in our findings. We concluded that guarana presents substances that have antitumoral effects in mice, enclosing a chemopreventive or antineoplastic effect depending on the model studied

Chemical carcinogenesis by DMBA (7,12-dimethylbenzanthracene) in female BALB/c mice: new facts / Carcinogenese quimica por DMBA (7,12-dimethylbenzanthracene) em camundongos femeas BALB/c: novos fatos

Polycyclic aromatic hydrocarbons are known carcinogens used in rodent experimental models. In this study, the carcinogen DMBA (7,12-dimethylbenzanthracene) was administered by gavage, diluted in corn oil, to female BALB / c mice at hebdomadary doses of 1 mg per animal for 1, 3, 6 or 9 weeks. Animals were weighed and monitored weekly until death. Remaining animals were euthanized at the age of 53 weeks. At necropsy, representative fragments of neoplasms were collected and routinely processed for histopathological analysis. Of all mice that received DMBA, 68.57% developed some type of tumor. Of the 70 mice treated with various doses of DMBA, 22 (31.43%) developed mammary tumors. The adenoacanthoma was the most commonly (18.75%) diagnosed histological type of breast cancer. Lung (15.71%), lymphoid tissue (11.43%), stomach (7.14%) and skin (2.86%) were also primary sites of tumor development. One third (33.33%) of the mice receiving 1 mg of DMBA developed lung cancer. Therefore, the administration of DMBA was shown to be an efficient model of carcinogenesis in mice, especially for the study of breast cancer, when using the highest dose, and lung, when using the lowest dose. Carcinogenesis models have been used for several purposes in cancer research. These results represent new facts for a classic carcinogenesis model.

Hidrocarbonetos policíclicos e aromáticos são carcinógenos usados em modelos experimentais em roedores. Neste estudo, o carcinógeno DMBA (7,12-dimetilbenzantraceno) foi administrado por gavagem, diluído em óleo de milho, para camundongos BALB/c em doses hebdomadárias de 1 mg por animal por 1, 3, 6 ou 9 semanas. Os animais foram pesados e monitorados semanalmente até a morte. Os animais remanescentes foram eutanasiados com a idade de 53 semanas. Na necroscopia, fragmentos representativos das neoplasias foram colhidos e rotineiramente processados para exame histopatológico. De todos os animais que receberam DMBA, 68,57% desenvolveram algum tipo de tumor. Entre os 70 camundongos tratados com diferentes doses de DMBA, 22 (31,43%) desenvolveram neoplasias mamárias. O adenoacantoma foi o tumor mamário mais comumente diagnosticado (18,75%). Pulmões (15,71%), tecido linfoide (11,43%), estômago (7,14%) e pele (2,86%) foram também locais primários de desenvolvimento de neoplasias. Um terço (33,33%) dos camundongos que receberam 1 mg de DMBA desenvolveram neoplasias pulmonares. Portanto, a administração de DMBA foi considerada um modelo eficiente de carcinogênese em camundongos, especialmente para o estudo de neoplasias mamárias, quando a maior dose é utilizada, e de neoplasias pulmonares, quando utilizada a menor dose. Os modelos de carcinogênese química têm sido usados para diversos est

Compensatory kidney hypertrophy/hyperplasia after nephrectomy in mice: alterations of connexin 43 (Cx43) phosphorylated isoforms

Compensatory kidney hypertrophy/hyperplasia leads to several changes in kidney structure and function, as increased glomeruli filtration. The aim of this study was to evaluate connexin 43 in remnant mouse kidneys after unilateral nephrectomy. The right kidney was surgically removed from BALB/c mice. Groups were euthanized at 24, 48 and 72 hours, and at 7 and 30 days. Kidney sections of the reminiscent kidneys were stained with Periodic Acid/Schiff and additional slides were submitted to BrdU and Cx43 immunohistochemistry. The results demonstrated an increase in kidney weight as early as 24 hours through 30 post-nephrectomy. In addition, BrdU positive epithelial cells increased at 24 and 48 hours post-nephrectomy. Cx43 was detected in the cytoplasm and membrane of epithelial cells and vasculature. Taking into consideration the quantity, intensity and localization of Cx43 immunostaining pattern, we observed that nephrectomized mice presented lower Cx43 expression and a cytoplasmic localization after 24 hours, peaking in 48 hours. Furthermore, western blot revealed that during the first 24 and 48 hours after nephrectomy, P0 (unphosphorylated) and P1 (phosphorylated) Cx43 disappeared, and the products of Cx43 degradation were reduced. On the other hand, after 72 hours the P0 and P1 state reappeared and the amount of degraded peptides also increased. Seven and thirty days after neph

Consensus for the diagnosis, prognosis and treatment of canine mammary tumors

The purpose of this paper is to establish criteria that could guide the diagnosis, prognosis and treatment of canine mammary neoplasias. It was elaborated during the Mammary Pathology Meeting: Diagnosis, Prognosis and Treatment of the Canine Mammary Neoplasm, held on November 6th and 7th, 2010 in Belo Horizonte MG, Brazil, sponsored by the Laboratory of Comparative Pathology UFMG, with the support of the Brazilian Association of Veterinary Pathology (ABPV) and Brazilian Association of Veterinary Oncology (ABROVET). Academics from several regions of Brazil were present and contributed to this work.