Aspectos clínicos, laboratoriais e tratamento da leucemia mieloide crônica em cão relato de caso / Clinical aspects, hematological changes and treatments of chronic myeloid leukemia in dog: case report

O objetivo deste relato é apresentar o caso clínico de uma cadela, sem raça definida, com cinco anos de idade, diagnosticada com leucemia mieloide crônica (LMC). As leucemias são neoplasias malignas que se originam de células precursoras da medula óssea e as consequências podem ser trombocitopenia, anemia, leucocitose persistente e presença de células neoplásicas no sangue. O tratamento de escolha envolve o uso de inibidores de tirosina quinase, porém este não pode ser usado neste caso. Dessa forma a cadela recebeu diferentes protocolos quimioterápicos que incluíram inicialmente hidroxiureia, citarabina, doxorrubicina e prednisona. Devido a remissão parcial dos sinais clínicos e a resposta terapêutica pouco duradoura a essas medicações o protocolo foi alterado para quimioterapia metronômica com clorambucil. O uso desses quimioterápicos não foram eficazes em reduzir a leucocitose e controlar a anemia e trombocitopenia da paciente, devido a ocorrência do surgimento de células imaturas no sangue e resistência aos quimioterápicos. Na ausência da crise e da possibilidade do uso dos inibidores de tirosina quinase, a hidroxiureia permanece sendo o quimioterápico de eleição. O animal apresentou sobrevida de 210 dias, devido a leucocitose e anemia severas pouco responsivas ao protocolo terapêutico utilizado e o surgimento no hemograma de precursores neutrofilicos que ocorreu 46 dias

The aim of this report is to present the clinical case of a five-year-old mixed breed female dog diagnosed with chronic myeloid leukemia (CML). Leukemias are malignant neoplasms that originate from bone marrow precursor cells and the consequences can be thrombocytopenia, anemia, persistent leukocytosis and the presence of neoplastic cells in the blood. The treatment of choice involves the use of tyrosine kinase inhibitors, but it cannot be used in this case. Thus, the dog received different chemotherapy protocols that initially included hydroxyurea, cytarabine, doxorubicin and prednisone. Due to the partial remission of clinical signs and the short-term therapeutic response to these medications, the protocol was changed to metronomic chemotherapy with chlorambucil. The use of these chemotherapeutic agents was not effective in reducing leukocytosis and controlling the patient’s anemia and thrombocytopenia, due to the occurrence of immature cells in the blood and resistance to chemotherapeutic agents. In the absence of the crisis and the possibility of using tyrosine kinase inhibitors, hydroxyurea remains the chemotherapy of choice. The animal had a 210-day survival, due to severe leukocytosis and anemia, which were not responsive to the therapeutic protocol used and the appearance in the blood count of neutrophilic precursors that occurred 46 days after the beginning of hydroxyurea treatment.

Aspectos clínicos, laboratoriais e tratamento da leucemia mieloide crônica em cão: relato de caso / Clinical aspects, hematological changes and treatments of chronic myeloid leukemia in dog: case report

O objetivo deste relato é apresentar o caso clínico de uma cadela, sem raça definida, com cinco anos de idade, diagnosticada com leucemia mieloide crônica (LMC). As leucemias são neoplasias malignas que se originam de células precursoras da medula óssea e as consequências podem ser trombocitopenia, anemia, leucocitose persistente e presença de células neoplásicas no sangue. O tratamento de escolha envolve o uso de inibidores de tirosina quinase, porém este não pode ser usado neste caso. Dessa forma a cadela recebeu diferentes protocolos quimioterápicos que incluíram inicialmente hidroxiureia, citarabina, doxorrubicina e prednisona. Devido a remissão parcial dos sinais clínicos e a resposta terapêutica pouco duradoura a essas medicações o protocolo foi alterado para quimioterapia metronômica com clorambucil. O uso desses quimioterápicos não foram eficazes em reduzir a leucocitose e controlar a anemia e trombocitopenia da paciente, devido a ocorrência do surgimento de células imaturas no sangue e resistência aos quimioterápicos. Na ausência da crise e da possibilidade do uso dos inibidores de tirosina quinase, a hidroxiureia permanece sendo o quimioterápico de eleição. O animal apresentou sobrevida de 210 dias, devido a leucocitose e anemia severas pouco responsivas ao protocolo terapêutico utilizado e o surgimento no hemograma de precursores neutrofilicos que ocorreu 46 dias após ao início do tratamento com hidroxiureia.

The aim of this report is to present the clinical case of a five-year-old mixed breed female dog diagnosed with chronic myeloid leukemia (CML). Leukemias are malignant neoplasms that originate from bone marrow precursor cells and the consequences can be thrombocytopenia, anemia, persistent leukocytosis and the presence of neoplastic cells in the blood. The treatment of choice involves the use of tyrosine kinase inhibitors, but it cannot be used in this case. Thus, the dog received different chemotherapy protocols that initially included hydroxyurea, cytarabine, doxorubicin and prednisone. Due to the partial remission of clinical signs and the short-term therapeutic response to these medications, the protocol was changed to metronomic chemotherapy with chlorambucil. The use of these chemotherapeutic agents was not effective in reducing leukocytosis and controlling the patient's anemia and thrombocytopenia, due to the occurrence of immature cells in the blood and resistance to chemotherapeutic agents. In the absence of the crisis and the possibility of using tyrosine kinase inhibitors, hydroxyurea remains the chemotherapy of choice. The animal had a 210-day survival, due to severe leukocytosis and anemia, which were not responsive to the therapeutic protocol used and the appearance in the blood count of neutrophilic precursors that occurred 46 days after the beginning of hydroxyurea treatment.

Detection molecular of Rangelia vitalii in dogs from Parana State, Southern Brazil / Detecção molecular de Rangelia vitalii em cães do Estado do Paraná, Sul do Brasil

Abstract Rangelia vitalii infects erythrocytes, leukocytes and endothelial cells of dogs. The present study aimed to report the molecular detection confirmed by sequencing of R. vitalii in the state of Paraná, as well as describe the clinical, hematological and biochemical alterations of the infected dogs. Three sick dogs from the metropolitan area of Curitiba, PR, Brazil, underwent a physical exam, and laboratory tests included hematology, biochemistry, polymerase chain reaction (PCR), and gene sequencing. Clinical signs included apathy, anorexia, and hemorrhage. Intra-erythrocytic and extracellular piroplasms were found on peripheral blood smears from all three dogs. Blood samples from these animals were positive for Babesia sp. by PCR targeting 18S rRNA. PCR products from all three dogs were sequenced, and BLAST analysis showed that the PCR-generated sequences were highly homologous with those of R. vitalii previously reported. Hematologic findings included severe anemia, shift of neutrophils to the regenerative left, and thrombocytopenia. Serum urea levels were increased in all three dogs, and direct bilirubin levels were elevated in one dog.

Resumo Rangelia vitalii infecta eritrócitos, leucócitos e células endoteliais de cães. O presente estudo objetivou relatar a detecção molecular confirmada por sequenciamento de R. vitalii no estado do Paraná e descrever as alterações clínicas, hematológicas e bioquímicas dos cães infectados. Três cães doentes da região metropolitana de Curitiba, PR, Brasil, foram submetidos a exame físico e exames laboratoriais que incluíram hematologia, bioquímica, reação em cadeia da polimerase (PCR) e sequenciamento genético. Os sinais clínicos incluíram apatia, anorexia e hemorragia. Piroplasmas intra-eritrocíticos e extracelulares foram encontrados em esfregaços de sangue periférico dos três cães. As amostras de sangue destes animais foram positivas para Babesia sp. pela PCR baseada no gene 18S rRNA. Os produtos de PCR dos três cães foram sequenciados e a análise de BLAST mostrou que as seqüências geradas por PCR eram altamente homólogas com as de R. vitalii previamente relatadas. Os achados hematológicos incluíram anemia grave, desvio de neutrófilos à esquerda regenerativo e trombocitopenia. Os níveis de uréia no soro aumentaram nos três cães, e os níveis de bilirrubina direta foram elevados em um cão.

Detection molecular of Rangelia vitalii in dogs from Parana State, Southern Brazil

Abstract Rangelia vitalii infects erythrocytes, leukocytes and endothelial cells of dogs. The present study aimed to report the molecular detection confirmed by sequencing of R. vitalii in the state of Paraná, as well as describe the clinical, hematological and biochemical alterations of the infected dogs. Three sick dogs from the metropolitan area of Curitiba, PR, Brazil, underwent a physical exam, and laboratory tests included hematology, biochemistry, polymerase chain reaction (PCR), and gene sequencing. Clinical signs included apathy, anorexia, and hemorrhage. Intra-erythrocytic and extracellular piroplasms were found on peripheral blood smears from all three dogs. Blood samples from these animals were positive for Babesia sp. by PCR targeting 18S rRNA. PCR products from all three dogs were sequenced, and BLAST analysis showed that the PCR-generated sequences were highly homologous with those of R. vitalii previously reported. Hematologic findings included severe anemia, shift of neutrophils to the regenerative left, and thrombocytopenia. Serum urea levels were increased in all three dogs, and direct bilirubin levels were elevated in one dog.

Resumo Rangelia vitalii infecta eritrócitos, leucócitos e células endoteliais de cães. O presente estudo objetivou relatar a detecção molecular confirmada por sequenciamento de R. vitalii no estado do Paraná e descrever as alterações clínicas, hematológicas e bioquímicas dos cães infectados. Três cães doentes da região metropolitana de Curitiba, PR, Brasil, foram submetidos a exame físico e exames laboratoriais que incluíram hematologia, bioquímica, reação em cadeia da polimerase (PCR) e sequenciamento genético. Os sinais clínicos incluíram apatia, anorexia e hemorragia. Piroplasmas intra-eritrocíticos e extracelulares foram encontrados em esfregaços de sangue periférico dos três cães. As amostras de sangue destes animais foram positivas para Babesia sp. pela PCR baseada no gene 18S rRNA. Os produtos de PCR dos três cães foram sequenciados e a análise de BLAST mostrou que as seqüências geradas por PCR eram altamente homólogas com as de R. vitalii previamente relatadas. Os achados hematológicos incluíram anemia grave, desvio de neutrófilos à esquerda regenerativo e trombocitopenia. Os níveis de uréia no soro aumentaram nos três cães, e os níveis de bilirrubina direta foram elevados em um cão.

Laminar lesions in horses with systemic oxidative stress, committed by experimentally induced or naturally occurring gastrointestinal disorders / Lesões laminares em cavalos com estresse oxidativo sistêmico, acometidos por doenças gastrintestinais experimentalmente induzidas ou de ocorrência natural

Laminitis in horses can be associated with lesions in multiple organs secondary to sepsis. Twenty-one horses suffering from gastrointestinal disorders were used in the experiment; 7 horses with experimentally induced endotoxemia and intestinal ischaemia, and 14 horses suffering from naturally occurring colic syndrome. Tissue samples of lungs, liver, heart, brain, cerebellum and hoof laminar tissue were collected for histopathological and oxidative stress evaluation using nitrotyrosine and superoxide dismutase (SOD2) immunostaining. The horses were divided into two groups: the non-oxidative lesions group (NOLG), with 7 horses showing weak immunostaining in lungs, liver and kidney, and the oxidative lesions group (OLG), with 14 horses showing immunostaining indicating systemic oxidative stress in multiple organs. The horses from OLG showed increase of laminar lesions and SOD2 immunostaining in multiple organs when compared to the horses from the NOLG. No differences were found ln regard to laminar immunostaining by nitrotyrosine and SOD2 between experimental groups. It was concluded that systemic oxidative stress can be associated with the development of laminar lesions, and that the laminar tissue does not respond to oxidative stress with increase of SOD as occurs in other organs.(AU)

A laminite em equinos pode estar associada à lesão em múltiplos órgãos secundária a sepse. Foram utilizados 21 cavalos com afecções gastrintestinais, sendo sete com endotoxemia e isquemia intestinal induzidos experimentalmente, e 14 cavalos com síndrome cólica de origem natural. Amostras teciduais de pulmão, rim, fígado, coração, cérebro e cerebelo e de tecido laminar do casco foram coletadas para avaliação de lesão histopatológica e estresse oxidativo, pela imunomarcação de nitrotirosina e superóxido dismutase (SOD2). Os animais foram divididos em dois grupos: grupo sem lesão oxidativa (NOLG), com sete cavalos com fraca imunomarcação em pulmão, fígado e rim, e grupo lesão oxidativa (OLG), contendo 14 cavalos com imunomarcação indicando estresse oxidativo em múltiplos órgãos. Os cavalos do grupo OLG apresentaram aumento de lesões laminares e imunomarcação para SOD2 em múltiplos órgãos, quando comparados ao NOLG. Não houve diferença sobre a imunomarcação laminar para nitrotirosina e SOD2 entre os grupos experimentais. Conclui-se que o estresse oxidativo sistêmico está associado ao desenvolvimento de lesões laminares, e que o tecido laminar não responde ao estresse oxidativo com aumento de SOD como ocorre nos outros órgãos.(AU)

Characterization of an alcoholic hepatic steatosis model induced by ethanol and high-fat diet in rats

Alcoholic liver disease is characterized by a wide spectrum of liver damage, which increases when ethanol is associated with high-fat diets (HFD). This work aimed to establish a model of alcoholic hepatic steatosis (AHS) by using a combination of 10% ethanol and sunflower seeds as the source of HFD. Male rats received water or 10% ethanol and regular chow diet and/or HFD, which consisted of sunflower seeds. The food consumption, liquid intake and body weight of the rats were monitored for 30 days. After this period, blood was collected for biochemical evaluation, and liver samples were collected for histological, mitochondrial enzyme activity and oxidative stress analyses. Our results indicated that the combination of 10% ethanol and HFD induced micro- and macrosteatosis and hepatocyte tumefaction, decreased the levels of reduced glutathione and glutathione S-transferase activity and increased the level of lipoperoxidation and superoxide dismutase activity. The mitochondrial oxidation of NADH and succinate were partially inhibited. Complexes I and II were the main inhibition sites. Hepatic steatosis was successfully induced after 4 weeks of the diet, and the liver function was modified. The combination of 10% ethanol and sunflower seeds as an HFD produced an inexpensive model to study AHS in rats.

Proteínas de fase aguda em cadelas com neoplasia mamária / Acute phase proteins in female dogs with mammary tumors

As proteínas de fase aguda (PFA) apresentam concentrações séricas alteradas mediante processos infecciosos, inflamatórios e neoplásicos. Objetivou-se com este trabalho avaliar as variações séricas das PFA em cadelas portadoras de neoplasia mamária, comparando com a avaliação histológica e leucograma. As PFA foram avaliadas em 45 cadelas com tumor de mama, distribuídas nos grupos neoplasia benigna (n=13), maligna não ulcerada (n=24) e maligna ulcerada (n=8). O grupo controle foi composto por 20 cadelas saudáveis. Foram realizados o teste de eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE) para identificar as PFA (albumina, ceruloplasmina, transferrina, haptoglobina Hp, α-1 antitripsina e α-1 glicoproteina ácida) e o teste ultrassensível para proteína C reativa (PCR). As pacientes com neoplasia mamária maligna ulcerada apresentaram elevações sérica para PCR e Hp e redução da albumina (P<0,05, One-Way ANOVA e Teste de Dunn). Nessas pacientes, foi observada correlação positiva entre o leucograma inflamatório e o aumento das PFA (P=0,002, Teste de Fisher) e não foram observadas correlações entre as PFA e os subtipos histológicos. Conclui-se que avaliações conjuntas da PCR, Hp e albumina podem ser utilizadas como ferramenta de auxílio diagnóstico e prognóstico em cadelas com neoplasia mamária.

Acute phase proteins (APPs) are serum proteins whose concentrations change after infectious and inflammatory disease, and cancer. The aims of this study were to evaluate changes in APPs concentration and to correlate these findings with histological classification and WBC in female dogs with mammary tumors. APPs were studied in 45 female dogs with mammary tumor distributed in the following groups: benign (n=13), malignant without tumor ulceration (n=24), and malignant with tumor ulceration (n=8). SDS-polyacrylamide gel (SDS-PAGE) electrophoresis was used to measure APPs concentrations (albumin, ceruloplasmin, transferrin, haptoglobinHp, α-1-acid glycoprotein and α-1-antitrypsin) and ultrasensitive assay was used to evaluate serum C-reactive protein (CRP). Patients with malignant mammary neoplasia plus ulceration had significant increase of CRP and Hp, and had decreased levels of albumin (P<0.05, One-Way ANOVA and Dunn Test). Positive correlation among APPs and inflammatory leukocytosis were observed (P=0.002, Fisher test). No correlation was observed between APPs and histological subtype. In conclusion, combined changes of CRP, Hp and albumin may be used as a prediagnostic tool and prognosis in dogs with mammary tumors.