Combination therapy of canine osteosarcoma with canine bone marrow stem cells, bone morphogenetic protein and carboplatin in an in vivo model.

Osteosarcoma (OSA) is the most common malignant bone cancer in children and dogs. The therapeutic protocols adopted for dogs and humans are very similar, involving surgical options such as amputation. Besides surgical options, radiotherapy and chemotherapy also are adopted. However, hematologic, gastrointestinal and renal toxicity may occur because of chemotherapy treatments. Recent study clearly showed that mesenchymal stem cells (MSCs) combined with recombinant human bone morphogenetic protein (rhBMP-2) may be associated with decreases of the tumorigenic potential of canine OSA. The aim of this study was to analyse the efficacy of chemotherapy with carboplatin and rhBMP-2 with MSCs in a canine OSA in vivo model. Canine OSA cells were implanted in mice Balb-c/nude with MSCs, rhBMP-2 and carboplatin. Flow cytometry and PCR for markers involved in tumour suppression pathways were analysed. Results showed that the combination of MSCs and rhBMP-2 reduced tumour mass and infiltration of neoplastic cells in tissues more efficiently than carboplatin alone. Thus it was demonstrated that the use of rhBMP-2 and MSCs, in combination with conventional antineoplastic, may be an efficient treatment strategy.

Bovine yolk sac: from morphology to metabolomic and proteomic profiles.

In several species, placentation involves the presence of two different membranes responsible for maternal-fetal exchanges: the yolk sac and the chorioallantoic placenta. The yolk sac plays important roles in embryonic survival, mainly during the early stages of gestation. In bovine, it is a transitional membrane that is present until day 50-70 of pregnancy. Herein, we evaluated the morphological and molecular aspects of the yolk sac of bovine embryos during 24 to 52 days of gestation. A total of 69 embryos were allocated into three groups according to the crown-rump length and estimated ages. Yolk sac samples were then subjected to morphological and molecular analysis using mass spectrometry and nuclear magnetic resonance techniques. In contrast to alanine, which was observed only in Group I, during all gestational stages, we identified important metabolites such as aspartate, taurine, glycerophosphocholine, creatinine, creatine, hydrouracil, glutamate, glutamine, lactate, lysine, valine, myo-inositol, cadaverine, and choline. In addition, 314 random sequences of proteins were identified in the bovine yolk sac, and 47 of these were considered to be specific. Changes in alpha-fetoprotein and carcinoembryonic antigen concentrations during gestation were also evaluated. In conclusion, the majority of these proteins are related to the development of secondary metabolites that are involved in the activation of other proteins and metabolites, and in signaling pathways that are responsible for maternal-fetal exchanges, activation of programmed cell death mechanisms, and cellular differentiation, and also in proteins that are responsible for the yolk sac involution that is required to establish chorioallantoic placentation.

Comparação da imunolocalização de receptores de progesterona em placentas de bovinos normais e clonados / Comparison of the imunolocalization of progesterone receptors in placentas of bovine normal and cloned

A progesterona é um hormônio fundamental na manutenção da gestação nos mamíferos. Em bovinos clonados, perdas gestacionais significativas ocorrem devido à deficiência placentária. Tendo em vista esse problema, o objetivo do trabalho foi analisar em diferentes idades gestacionais a imunolocalização para receptores de progesterona em placentas de animais não clonados e clonados. Tecidos placentomais de bovinos não clonados foram obtidos da linha de abate no Matadouro Mantiqueira, enquanto os tecidos placentomais dos animais clonados com idades gestacionais de 68, 135, 225 e 270 dias de gestação foram obtidos em empresas especializadas em clonagem animal. Os tecidos foram processados e embebidos em paraplast para localização de receptores de progesterona, utilizando-se a técnica de imuno-histoquímica pelo anticorpo monoclonal (Progesterone receptor SER 294). Pela imuno-histoquímica, comparando-se a concentração de receptores de progesterona na placenta de animais clonados com as de animais não clonados, em diferentes idades gestacionais, observou-se que as células dos placentomas dos animais clonados apresentaram uma marcação mais intensa de receptor de progesterona nas idades de 135 e 270 dias de gestação em relação aos não clonados. Tal fato sugere que a presença de progesterona no final da gestação pode ser mais uma das possíveis causas de gestações prolongadas.

To progesterone is a fundamental hormone in the maintenance of the gestation in the mammals. In bovine they cloned, losses gestational significant, occur due to the placental deficiency. Having in mind that problem, the objective of the work was analyze in different gestational ages to immune stained for receivers of progesterone in placentas of normal animals and cloned. Placentomal tissues of bovine normal were obtained of the line of counts on in the Slaughterhouse Mantiqueira, while the placentomal tissues of the animals cloned with gestational ages of 68, 135, 225 and 270 days of gestation were obtained in companies specialized in animal cloning. The fabrics were prosecuted and absorbed in paraplast for progesterone receivers location, utilizing itself to technical of immunohistochemistry by the antibody monoclonal (Progesterone receiver BE 294). By the immunohistochemistry comparing the progesterone receivers concentration in the placenta of animals cloned, with the of animals done not manipulate, in different gestational ages we observe that the cells of the placentomal tissues of the animals manipulated (cloned) presented a more intense marking of receiver of progesterone in the ages of 135 and 270 days of gestation regarding the done not manipulate. Suggesting that the presence of progesterone in the end of the gestation can be more one of the possible cause of gestations prolonged.

Comparação da imunolocalização de receptores de progesterona em placentas de bovinos normais e clonados / Comparison of the imunolocalization of progesterone receptors in placentas of bovine normal and cloned

A progesterona é um hormônio fundamental na manutenção da gestação nos mamíferos. Em bovinos clonados, perdas gestacionais significativas ocorrem devido à deficiência placentária. Tendo em vista esse problema, o objetivo do trabalho foi analisar em diferentes idades gestacionais a imunolocalização para receptores de progesterona em placentas de animais não clonados e clonados. Tecidos placentomais de bovinos não clonados foram obtidos da linha de abate no Matadouro Mantiqueira, enquanto os tecidos placentomais dos animais clonados com idades gestacionais de 68, 135, 225 e 270 dias de gestação foram obtidos em empresas especializadas em clonagem animal. Os tecidos foram processados e embebidos em paraplast para localização de receptores de progesterona, utilizando-se a técnica de imuno-histoquímica pelo anticorpo monoclonal (Progesterone receptor SER 294). Pela imuno-histoquímica, comparando-se a concentração de receptores de progesterona na placenta de animais clonados com as de animais não clonados, em diferentes idades gestacionais, observou-se que as células dos placentomas dos animais clonados apresentaram uma marcação mais intensa de receptor de progesterona nas idades de 135 e 270 dias de gestação em relação aos não clonados. Tal fato sugere que a presença de progesterona no final da gestação pode ser mais uma das possíveis causas de gestações prolongadas.(AU)

To progesterone is a fundamental hormone in the maintenance of the gestation in the mammals. In bovine they cloned, losses gestational significant, occur due to the placental deficiency. Having in mind that problem, the objective of the work was analyze in different gestational ages to immune stained for receivers of progesterone in placentas of normal animals and cloned. Placentomal tissues of bovine normal were obtained of the line of counts on in the Slaughterhouse Mantiqueira, while the placentomal tissues of the animals cloned with gestational ages of 68, 135, 225 and 270 days of gestation were obtained in companies specialized in animal cloning. The fabrics were prosecuted and absorbed in paraplast for progesterone receivers location, utilizing itself to technical of immunohistochemistry by the antibody monoclonal (Progesterone receiver BE 294). By the immunohistochemistry comparing the progesterone receivers concentration in the placenta of animals cloned, with the of animals done not manipulate, in different gestational ages we observe that the cells of the placentomal tissues of the animals manipulated (cloned) presented a more intense marking of receiver of progesterone in the ages of 135 and 270 days of gestation regarding the done not manipulate. Suggesting that the presence of progesterone in the end of the gestation can be more one of the possible cause of gestations prolonged.(AU)

Ossificação endocondral em embriões e fetos de bovinos / Endocondral ossification in bovine embryos and fetus

Estudou-se a ossificação endocontral de 18 embriões e 12 fetos de até três meses de gestação, os quais foram coletados de úteros gestantes em frigoríficos e abatedouros. Os úteros foram dissecados e, em seguida, realizou-se uma incisão dorsal até o cérvix para avaliações macroscópicas dos embriões e fetos. Para o estudo microscópico foram realizadas técnicas de inclusão, seguidas de marcação dos depósitos de cálcio e fósforo, responsável pela ossificação dos moldes de cartilagem. Foram identificados hipertrofia da cartilagem e morte dos condrócitos e aumento da área de depósito de cálcio e fósforo, por volta da 10ª semana gestacional (74 dias). Durante a 11ª semana de gestação (81 dias), os grupamentos de carbonato de cálcio e fósforo espalharam-se por todo o osso, sendo mais intenso na diáfise.(AU)

The endocondral ossification process was analyzed in 18 embryos and 12 fetus until three months of pregnancy, which had been collected from pregnant uterus at slaughterhouse. Later, the uterus was separated followed by a dorsal incision from the cervix for better gross evaluations of the embryos and fetus. For the microscopical study, histological techniques had been performed followed by staining of the deposits of calcium and phosphorus match, responsible for the ossification of the cartilage casts, used as Von Kossa stain. Cartilage hypertrophy and death of chondrocytes, enlargement of the area of calcium and phosphorus deposit were foundon the 10th week of pregnancy (74 d). During the 11th week of pregnancy (81 d), the calcium carbonate and phosphorus had spread to all bones, being more intense in diaphysis.(AU)

Ossificação endocondral em embriões e fetos de bovinos / Endocondral ossification in bovine embryos and fetus

Estudou-se a ossificação endocontral de 18 embriões e 12 fetos de até três meses de gestação, os quais foram coletados de úteros gestantes em frigoríficos e abatedouros. Os úteros foram dissecados e, em seguida, realizou-se uma incisão dorsal até o cérvix para avaliações macroscópicas dos embriões e fetos. Para o estudo microscópico foram realizadas técnicas de inclusão, seguidas de marcação dos depósitos de cálcio e fósforo, responsável pela ossificação dos moldes de cartilagem. Foram identificados hipertrofia da cartilagem e morte dos condrócitos e aumento da área de depósito de cálcio e fósforo, por volta da 10ª semana gestacional (74 dias). Durante a 11ª semana de gestação (81 dias), os grupamentos de carbonato de cálcio e fósforo espalharam-se por todo o osso, sendo mais intenso na diáfise.

The endocondral ossification process was analyzed in 18 embryos and 12 fetus until three months of pregnancy, which had been collected from pregnant uterus at slaughterhouse. Later, the uterus was separated followed by a dorsal incision from the cervix for better gross evaluations of the embryos and fetus. For the microscopical study, histological techniques had been performed followed by staining of the deposits of calcium and phosphorus match, responsible for the ossification of the cartilage casts, used as Von Kossa stain. Cartilage hypertrophy and death of chondrocytes, enlargement of the area of calcium and phosphorus deposit were foundon the 10th week of pregnancy (74 d). During the 11th week of pregnancy (81 d), the calcium carbonate and phosphorus had spread to all bones, being more intense in diaphysis.

Gastric precancerous lesions and Helicobacter pylori infection in relatives of gastric cancer patients from Northeastern Brazil.

BACKGROUND: Helicobacter pylori infection predisposes to gastric cancer. First-degree relatives of patients with gastric cancer have an increased risk of developing the disease. AIM: To evaluate the prevalence of gastric precancerous gastric lesions and H. pylori infection in first-degree relatives of non-cardia gastric cancer patients. METHODS: Gastric cancer relatives (n = 104) from a region with high prevalence of H. pylori infection and gastric cancer were invited for screening endoscopy; 80% of them had dyspeptic symptoms. The control group was composed of patients (n = 118) who concurrently underwent upper gastrointestinal endoscopy for investigation of dyspepsia with no family history of gastric cancer. The groups were matched for gender, age and social class. H. pylori status was evaluated by urease test, and histology and histological parameters were assessed according to the Houston Updated Sydney System. RESULTS: The prevalence of H. pylori infection was high in both the relative (84.7%) and the control (75.9%) groups. Corpus-predominant gastritis was more frequently observed in the relative group, whereas antral gastritis predominated in the controls. The density of lymphoid follicles was higher among the relatives. Also, intestinal metaplasia in the corpus and dysplasia were more prevalent in the cancer relative group than in the control group. Early gastric cancer was detected in 1 relative of gastric cancer patient with high-grade dysplasia. CONCLUSION: Individuals with a family history of non-cardia gastric cancer need to be routinely screened for H. pylori infection and precancerous gastric lesions.

Human neurobrucellosis with intracerebral granuloma caused by a marine mammal Brucella spp.

We present the first report of community-acquired human infections with marine mammal-associated Brucella spp. and describe the identification of these strains in two patients with neurobrucellosis and intracerebral granulomas. The identification of these isolates as marine mammal strains was based on omp2a sequence and amplification of the region flanking bp26.