Made available in DSpace on 2014-10-09T12:28:32Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:56:01Z (GMT). No. of bitstreams: 0 / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Orientador: Helio Langoni / Resumo: Mycoplasma spp., tem distribuição mundial e é um patógeno relevante em medicina veterinária. As mastites causadas por Mycoplasma spp. mais frequentes em grandes rebanhos leiteiros, porém este patógeno é subestimado no Brasil, onde se têm poucos relatos como agente causador de mastite, o que se deve possivelmente a quantidade reduzida de laboratórios que inclui a análise de M. bovis em sua rotina. Devido a necessidade de meios seletivos e de condições especiais para o seu isolamento. Diante disso um dos objetivos do presente estudo foi pesquisar a participação de Mycoplasma bovis na etiologia das mastites clínicas em amostras de leite de vacas de propriedades leiteiras de sete estados do Brasil, totalizando 561 amostras de leite, que foram cultivas em meio Hayflick adicionado de acetato de tálio a 0,01%, incubadas em ambiente de microaerofilia com 5% de CO2. Foram também submetidas a reação em cadeia da polimerase (PCR) para detecção de Mycoplasma spp. e Mycoplasma bovis. Pesquisouse também a microbiota aeróbica envolvida nas mastites, cultivando-se as mesmas amostras de leite em meios de ágar sangue bovino 5% e ágar MacConkey resultando 225 amostras positivas. Obtiveram-se 11 (1,96%) amostras positivas para Mycoplasma spp., no exame microbiológico, enquanto na detecção molecular obteve-se um total de 17 (3,03%) amostras positivas para Mycoplasma bovis. Pode-se concluir com os resultados obtidos pela presença e dispersão do Mycoplasma bovis nos rebanhos leiteiros avaliados, ap... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

The aim of this work was to gather and evaluate the evidence about the behaviour of dairy cows during the milking by a robot (including physiological reactions running after the milking). The following parameters were observed: number of milking per 24 hours, the time from onset to the robot and the teat cup deployment, the time between milking, the total time of milking, milk yield with each milking, average and total milk yield per day. The results were measured separately in heifers and in cows on the second and subsequent lactations. In the same time, there was also observed the behaviour of cows after leaving the milking robot - feed and water intake, physical activity and the rest for the time of 30 minutes after the milking. Simultaneously, there was also observed the effect of milking technology for the health of the cows, focused on diseases of the mammary gland. There were three ethologic observations which always lasted 24 hours. In addition to the observed data, there were evaluated the data records obtained from the milking machine (e. g. milk yield per one milking, the interval between single milkings, etc.). There were observed an average of 60 pieces of cows. Throughout the watching the cows in the barn behaved calmly and contentedly. They came to the milking machine spontaneously, only a few cases had to be herded for milking by the caregiver. However, they were all the heifers which did not have the sufficient experience with the milking. The counting of teat cup deployment attempts had to be abandoned because of the close proximity of the observer at the milking robot. In this case, the cows were not quiet and did not want to go in the robot spontaneously. The mammary gland disease was diagnosed at an average of 6.9 pieces of milked cows per one month. This high number of cases is ascribed to a poor hygiene of the stables and a poorer quality of feeding. This work was created in cooperation with the grants NAZV QJ1210144 and NAZV QJ1530058.

Submitted by Diogo Misoguti (diogo.misoguti@gmail.com) on 2016-06-30T17:50:23Z No. of bitstreams: 1 cp111463.pdf: 4432662 bytes, checksum: a585b229b3b920b2ee4fda25aa3206c7 (MD5) / Approved for entry into archive by Diogo Misoguti (diogo.misoguti@gmail.com) on 2016-06-30T17:52:42Z (GMT) No. of bitstreams: 1 cp111463.pdf: 4432662 bytes, checksum: a585b229b3b920b2ee4fda25aa3206c7 (MD5) / Made available in DSpace on 2016-06-30T17:52:42Z (GMT). No. of bitstreams: 1 cp111463.pdf: 4432662 bytes, checksum: a585b229b3b920b2ee4fda25aa3206c7 (MD5) Previous issue date: 2009 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / OBJETIVO: Este estudo teve como objetivo verificar as modificações causadas na proliferação celular e na expressão gênica da glândula mamária de ratas com 21, 35, 50 e 100 dias que foram expostas à 2,3,7,8 tetraclorodibenzo-p-dioxina (TCDD) durante a gestação (tratamento pré-natal) ou durante a lactação (tratamento pré-pubertal). MÉTODOS: Para o tratamento pré-natal, ratas Sprague-Dawley prenhas receberam no 15º dia após a concepção dose única de 3,0µg de TCDD/kg através de gavagem. Para o tratamento pré-pubertal, ratas lactantes receberam no 14º e 17º dias pós-parto 6,67ng ou 20,0ng de TCDD/g. Animais controles receberam volume equivalente de óleo de gergelim. Ao atingiram 21, 35, 50 e 100 dias de idade, as filhotes fêmeas das ratas que receberam TCDD foram sacrificadas e tiveram as glândulas mamárias extraídas para estudo do índice de proliferação celular e expressão gênica. Para o cálculo de índice de proliferação celular, dez ratas por grupo receberam uma injeção intraperitoneal de 200 mg/kg de 5-bromo-2’-deoxiuridina (BrdU) e através de imunohistoquímica foram identificadas as células que estavam proliferando em cada estrutura da glândula mamária. Outras dez ratas por grupo tiveram o RNA da glândula mamária extraído, e este foi utilizado para o estudo de expressão gênica através de microarrays. Para determinar os genes diferencialmente expressos utilizou-se o teste-t bayesiano empírico moderado. Genes diferencialmente expressos (p<0,01) foram então categorizados de acordo com a função biológica ou via canônica de ação. Ainda, genes que se destacaram foram validados através de RT-PCR em tempo real. RESULTADOS: O tratamento pré-pubertal alterou a proliferação celular nas glândulas mamárias das ratas com 35, 50 e 100 dias, principalmente nos botões terminais (TEBs). A análise da expressão gênica demonstrou que o número máximo de genes diferencialmente expressos foi observado nos animais com 100 dias que receberam TCDD durante a gestação. No tratamento pré-pubertal, observou-se que a alta dose causou mais alterações do que a dose baixa. Cyp1b1 foi o gene que mais sofreu aumento de expressão em diferentes idades. A categorização dos genes desregulados de acordo com a função biológica mostrou que TCDD tem uma ampla ação, principalmente após o tratamento pré-pubertal com maior dose. As classes de genes mais afetadas foram proto-oncogenes, genes supressores de tumor e genes relacionados com o metabolismo de lipídeos. A via de sinalização mais atingida foi a via de sinalização do receptor de aril-hidrocarbono (AhR). Também se observou alterações na expressão de genes participantes da via de metabolização do estradiol, aumentando a produção de metabólitos prejudiciais ao DNA. CONCLUSÕES: O tratamento com TCDD resultou em mudanças na proliferação celular e na expressão gênica da glândula mamária das ratas. As mudanças gênicas observadas indicam que a célula mamária é submetida a estímulos genotóxicos ao mesmo tempo em que genes de reparo de DNA e supressores do crescimento tumoral ficam ativados no intuito de manter a estabilidade genômica. / PURPOSE: This work studied the modifications in cell proliferation and gene expression in the rat mammary gland of animals 21, 35, 50 and 100 days old that were exposed to 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) during pregnancy (prenatal treatment) or during lactation (prepubertal treatment). METHODS: For the prenatal treatment, pregnant Sprague-Dawley rats received on the 15th day post-conception, single dose of 3.0µg TCDD/kg through gavage. For the prepubertal treatment, lactate dams received on the 14th and 17th days post-delivery either 6.67ng or 20.0ng TCDD/g. Control group received equivalent volume of sesame oil. When the female offspring reached 21, 35, 50 and 100 days of age, they were sacrificed and their mammary gland extracted for cell proliferation and gene expression analyses. To calculate the cell proliferation index, ten rats per group received an intraperitoneal injection of 200 mg/kg of 5-Bromo-2'- deoxyuridine (BrdU) and through immunohystochemistry the cells in proliferation were identified in each mammary gland structure. Other ten rats per group had the RNA extracted from the mammary gland and used for gene expression analysis through microarrays. To determine the differentially expressed genes empirical Bayes moderated one sample t-test was performed. Differentially expressed genes (p<0.01) were categorized accordingly with biological function and canonical pathways. In addition, important genes were validated through real time RT-PCR. RESULTS: The prepubertal treatment modified the cell proliferation of 35, 50 and 100 days-old rat mammary gland, mainly in the terminal end buds (TEBs). The gene expression analysis showed that the maximum number of differentially expressed genes was reached in 100 days-old rats that received TCDD during pregnancy. The prepubertal treatment induced more changes with higher dose of TCDD. Cyp1b1 was the gene the most up-regulated genes at different ages. The categorization of the dysregulated genes according with their biological function showed that TCDD has a wide action, especially after the prepubertal treatment with high dose. The groups of genes more affected by TCDD were the proto-oncogenes, tumor suppressor genes and genes related to the metabolism of lipids. The most affected canonical pathway was the aryl hydrocarbon receptor signaling pathway. In addition, genes related to estradiol metabolism were modulated increasing the production of metabolites that damage the DNA. CONCLUSIONS: The TCDD treatment induced changes in the cell proliferation and gene expression of the rat mammary gland. The genomic changes observed indicate that the mammary cell is submitted to genotoxic stimuli and DNA repair genes as tumor suppressor genes are activated to keep the genomic stability. / CAPES: BEX 2829/05-3

O colostro é fonte de energia e imunidade aos leitões neonatos. Para que a ingestão de colostro seja satisfatória, de modo a garantir a sobrevivência e ganho de peso dos leitões, as porcas devem produzir colostro suficiente para suprir as necessidades de toda a leitegada. O objetivo deste estudo foi avaliar fatores relacionados à fêmea, à leitegada e ao trabalho de parto que poderiam influenciar a produção de colostro em suínos. Foram utilizadas 96 matrizes suínas Camborough 25® com ordem de parto 1 a 7 e parição espontânea. As fêmeas e as leitegadas foram acompanhadas até 24 h após o início do parto. A produção de colostro foi estimada pela soma do consumo individual dos leitões, baseado no ganho de peso durante o primeiro dia de vida. O modelo de regressão múltipla explicou 28% da produção de colostro, sendo 24% explicados pelo peso total dos leitões nascidos vivos e 4% pela largura do primeiro par de tetos. O peso total dos leitões nascidos vivos foi correlacionado com o número total de leitões nascidos (r= 0,73) e nascidos vivos (r= 0,83). Quando separadas em duas classes de produção de colostro (ALTAPCOL; >3,4 kg; n = 50 vs BAIXAPCOL; ≤3,4 kg; n = 46), as fêmeas BAIXAPCOL tiveram menor número de leitões nascidos vivos e menor peso da leitegada viva (P<0,05). Por análise de regressão logística, foi verificado que fêmeas de OP 1, 2 e >3 apresentaram maior chance (P≤0,05) de estar no grupo BAIXAPCOL do que as fêmeas de OP 3. Fêmeas com mais de uma intervenção obstétrica no parto tiveram maior chance (P<0,05) de serem fêmeas BAIXAPCOL, em comparação ao grupo de fêmeas sem intervenções no parto. Este estudo evidenciou que o fator que mais influencia a produção de colostro é o peso total da leitegada viva, indiretamente representando o número de leitões amamentados pela porca. / Colostrum provides newborn piglets with energy and with passive immunity. An adequate colostrum intake, in order to fulfill the needs of piglets and then ensure their survival and weight gain, depends on sow’s ability to produce enough colostrum for the whole litter. The aim of this study was to evaluate factors involved on colostrum yield variability related to the sow, the litter and farrowing process. The experiment was conducted with 96 Camborough 25® sows of parities one to seven whose farrowing was spontaneous. Sows and their litters were followed until 24 h after farrowing onset. Colostrum production was estimated by summing up colostrum intake of each piglet of the litter. Colostrum ingestion by individual piglets was estimated using piglet weight gain during the first 24 h of life. The multiple regression model explained 28% of variation in colostrum yield, with 24% and 4% of variation being explained by the litter weight at birth and the width of first mammary glands, respectively. Litter weight at birth was positively correlated with the number of total born (r = 0.73) and liveborn piglets (r = 0.83).When separated into two classes of colostrum yield (HIGHPROD; >3.4 kg; n= 50 vs LOWPROD; ≤3.4 kg; n= 46), LOWPROD sows had lighter litters and fewer total born and liveborn piglets (P < 0.05). The logistic regression analysis showed that sows from parities 1, 2 and >3 had greater odds (P ≤ 0.05) to be in the LOWPROD group than parity 3. Sows with two or more obstetrical interventions had higher odds (P < 0.05) of belonging to the LOWPROD group than sows without interventions at farrowing. This study showed that litter weight at birth is the most important factor involved in colostrum yield variability, indirectly representing the number of piglets nursed by the sow.

The adult mammary gland is comprised of a bi-layered epithelium of luminal and myoepithelial cells surrounded by an adipocyte-rich fat pad, a highly collagenous extra-cellular matrix (ECM) and a number of other stromal and endothelial cell types. Mammary stem cells (MaSCs) reside within the epithelium and these are capable of repopulating a mammary fat pad that is devoid of epithelium, upon transplantation. It was sought to recapitulate this process of MaSCs repopulating a fat pad using a synthetic fat pad, engineered from a collagen scaffold invested with adipocytes, to provide an in vitro 3D model. Fluorescently tagged murine Axin2-expressing cells were obtained from transgenic mice and seeded into these scaffolds and cultured, mimicking the process of fat pad repopulation. Immunohistochemical analysis demonstrated that Axin2+ myoepithelial cells were rarely capable of forming bi-layered structures that expressed correct myoepithelial localisation and resemblance to a luminal morphology. Breast tumours surrounded by anisotropic (directional) collagen fibres running perpendicular to the tumour boundary are more aggressive and associated with poor patient prognosis. To recapitulate this anisotropic collagen phenotype in vitro, an ice-templating technique was used to modify the structure of the collagen scaffolds producing both non-directional (isotropic) and anisotropic internal architectures. Tumour cells from various breast cancer cell lines were seeded into both isotropic and anisotropic scaffolds to investigate whether this approach could distinguish cell type-specific migratory ability and whether anisotropy affected migration efficiency. Following analysis by confocal microscopy and ImageJ, anisotropic scaffolds were observed to enhance the migratory potential of MDA-MB-231 breast cancer cells. These results highlight the importance of collagen alignment and provide a reproducible method to quantitatively measure cell migration in 3D for cells derived from different breast cancer subtypes. Building on these data, the protocol was adapted to permit the direct investigation of tumour biopsy material. Given the heterogeneity of breast tumours, it was considered important to maintain tumour architecture and stromal components. Thus, murine mammary tumour fragments from two different established mammary cancer models were utilised and cultured in anisotropic collagen scaffolds in the presence or absence of adipocytes to allow an investigation of their influence on tumour cell migration. Further experiments included addition of various therapeutic drugs followed by immunofluorescence microscopy coupled with an optical clearing technique. These data demonstrated the utility of the model in determining both the rate and capacity of tumour cells to migrate through the engineered stroma while shedding light also on the mode of migration. Moreover, the response of different mammary tumour types to chemotherapeutic drugs could be could be readily quantified. To humanize the fat pad for subsequent human tissue analysis, human mesenchymal stem cells (MSC) were obtained from reduction mammoplasties and immortalised, before differentiating them into adipocytes within anisotropic collagen scaffolds. Human breast cancer cells were fluorescently tagged for tracking using lentiviral methods and were seeded into scaffolds invested with differentiated MSCs. Both cell types were successfully co-cultured for 7 days and imaged using multiphoton methods.

Submitted by FLÁVIA REGINA MORAES DA SILVA null (flavisbio@hotmail.com) on 2016-03-11T17:40:29Z No. of bitstreams: 1 Doutorado em Patologia (Flávia Regina Moraes da Silva) Versão Final.pdf: 2724811 bytes, checksum: a8ca539bbeff30c4bfa3fa6c2f9dfeba (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-03-14T17:46:13Z (GMT) No. of bitstreams: 1 silva_fm_dr_bot.pdf: 2724811 bytes, checksum: a8ca539bbeff30c4bfa3fa6c2f9dfeba (MD5) / Made available in DSpace on 2016-03-14T17:46:13Z (GMT). No. of bitstreams: 1 silva_fm_dr_bot.pdf: 2724811 bytes, checksum: a8ca539bbeff30c4bfa3fa6c2f9dfeba (MD5) Previous issue date: 2016-02-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O presente estudo teve como objetivo avaliar se a ingestão de dietas com deficiência ou suplementação de zinco, durante as fases de gestação, lactação e juvenil, interfere no desenvolvimento das glândulas mamárias e na susceptibilidade ao desenvolvimento de carcinogênese mamária induzida pela 7,12-dimetilbenz(a)antraceno (DMBA) em ratas da linhagem Sprague-Dawley (SD). Duas gerações, mães e prole de fêmeas receberam dietas com níveis adequados (35mg/Kg dieta), deficientes (3mg/Kg dieta) ou com suplementação (180mg/Kg dieta) de zinco durante a gestação e lactação. Após o desmame, as proles de fêmeas receberam as mesmas dietas das mães até o dia pós-natal (DPN) 51 ou 53. No DPN 51 fêmeas SD foram eutanasiadas para remoção das glândulas mamárias abdominais (D e E) ou receberam dose única de DMBA (50mg/kg; i.g.) para iniciação da carcinogênese e eutanasiadas no DPN 53 ou DPN 180. No dia DPN 53 as glândulas foram processadas para análises histológicas e imunoistoquímicas, e avaliação da expressão de genes relacionados a dano e reparo de DNA, apoptose, ciclo celular e sinalização de genes relacionados ao receptor de estrógeno e p53, por RT-qPCR com sistema Taqman Low density array (TLDA). Os tumores coletados ao longo do experimento e no DPN 180 foram processados para avaliação histológica. As fêmeas alimentadas com dieta deficiente de zinco apresentaram redução significativa no peso corpóreo nos DPNs 0, 10, 21 e 51. O número médio de brotos terminais (TEBs), ductos terminais (TEDs) e lóbulos alveolares (ABLs) mamários não diferiram entre as proles dos três grupos experimentais. No DPN 53, os índices de proliferação nas células epiteliais mamárias foram significativamente maiores no grupo que recebeu suplementação de zinco em relação ao grupo controle, enquanto os índices de células epiteliais em apoptose e RE-α positivas não diferiram entre os grupos. Além disso, a suplementação de zinco reduziu significativamente a expressão dos genes Api5 e Ercc1, que atuam como inibidor de apoptose e reparo de DNA, respectivamente. No DPN 180, tanto a deficiência quanto a suplementação de zinco na dieta não alteraram a latência, incidência, multiplicidade ou volume dos tumores mamários induzidos pela DMBA em relação ao grupo controle. No entanto, a suplementação de zinco aumentou o número total de tumores em 72% quando comparado aos demais grupos. Portanto a suplementação de zinco durante as fases iniciais da vida e perído juvenil aumentou marginalmente a suscetibilidade ao desenvolvimento de tumores mamários. / The present study was designed to evaluate whether dietary intake with zinc deficiency or supplementation during the pregnancy, lactation and juvenile stages interfere the development of mammary glands and susceptibility to the mammary carcinogenesis induced by 7,12-dimethylbenzanthracene (DMBA) in female rats Sprague-Dawley (SD) rats. Two generations, dams and female offspring received dietary with zinc levels adequate (35 mg/kg diet), deficiency (3 mg/kg diet) or supplementation (180 mg/kg diet) during pregnancy and lactation. After weaning, the females offspring received the same diets as their dams until post natal day (PND) 51 or 53. On PND 51, females SD were euthanized for removal of the mammary glands (right and left) or received a single dose of DMBA (50mg/kg, ig) for initiation of mammary carcinogenesis and euthanized on PND 53 or PND 180. On PND 53, mammary glands were processed for histological and immunohistochemical analyses, as well as evaluation of the expression of genes related to DNA damage and repair, apoptosis, cell cycle and signaling related to estrogen receptor and p53 by RT-qPCR with Taqman Low density array system (TLDA). Tumors collected throughout experiment or on PND 180 were processed for histological evaluation. The females fed with dietary zinc deficiency presented a significant reduction in body weight on PND 0, 10, 21 e 51. The mean number of the terminal buds (TEBs), terminal ducts (TEDs) and alveolar lobe (ABLs) in mammary gland did not differ among offspring of the three groups. On PND 53 the indexes of proliferation in the mammary glands epithelial cells were significantly higher in the zinc supplementation group in relation to the control group, while the indexes of apoptosis epithelial cell and positive ER-α did not differ among the groups. Also, dietary zinc supplementation significantly reduced the expression of the Api5 and Ercc1 genes related to apoptosis inhibitor and DNA repair, respectively. On PND 180 both dietary zinc deficiency as supplementation did not change the, latency, incidence, multiplicity or volume of DMBA-induced mammary tumors in relation to the control group. However, dietary zinc supplementation increased the total number of tumors in 72% compared to others groups. Therefore zinc supplementation during the early life and juvenile period marginally increased the susceptibility to the mammary tumors development.

Made available in DSpace on 2014-06-11T19:26:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-08-12Bitstream added on 2014-06-13T18:53:59Z : No. of bitstreams: 1 gelaleti_gb_me_sjrp.pdf: 706621 bytes, checksum: f280ddfe72d9aced3754ba3a7a380f9c (MD5) / O estudo dos tumores mamários em cadelas revela-se um excelente modelo para a investigação clínico-patológica, diagnóstica e prognóstica de neoplasias mamárias. As interleucinas desempenham um papel fundamental no câncer, em especial a interleucina-8 (IL-8), que possui propriedades tumorigênicas e pró-angiogênicas e a interleucina-12 (IL-12), com propriedades anti-metastáticas e anti-angiogênicas. O marcador tumoral antígeno-carboidrato (CA 15-3) tem importante significado clínico no seguimento de pacientes com neoplasia mamária. Além disso, o DNA livre circulante de células tem sido considerado um candidato a biomarcador para tumores. Assim, foram objetivos deste estudo mensurar os níveis séricos das proteínas IL-8, IL-12, CA 15-3, bem como estimar o número de cópias das sequências CAN SINEs relacionando-os aos parâmetros clínico-patológicos e a sobrevida do grupo a fim de inferir o possível valor prognóstico desses marcadores para o câncer de mama. Através do ensaio imunoenzimático e da técnica de PCR em tempo real foram avaliadas 33 cadelas com neoplasia mamária e 50 cadelas controle e suas concentrações foram estatisticamente relacionadas aos parâmetros estabelecidos. Foi encontrada diferença significante entre os níveis séricos de IL-8 nas cadelas com neoplasia mamária quando comparadas ao grupo controle e além disso, verificou-se aumento de IL-8 em relação à evolução tumoral, ao envolvimento linfonodal, recidiva e óbito. Baixos níveis de IL-12 foram observados em cadelas com tumores com maior tempo de evolução, cadelas com idade superior a 10 anos e, além disso, a alta taxa de sobrevida teve correlação com níveis elevados de IL-12. Assim, o aumento da IL-8 e a diminuição da IL-12 podem ser considerados marcadores prognósticos independentes confirmando seu papel como citocina... / The study of mammary tumors in dogs appears to be an excellent model to investigate the clinical and pathological diagnosis and prognosis of breast cancer. Interleukins play a key role in cancer, particularly interleukin-8 (IL-8), which have tumorigenic and pro-angiogenic properties and interleukin-12 (IL-12), with anti-metastatic and anti-angiogenic properties. The tumor marker carbohydrate antigen (CA 15-3) also has important clinical significance in monitoring patients with breast cancer. In addition, free circulating DNA has been considered a candidate biomarker for cancer. The objectives of this study were to measure serum levels of protein IL-8, IL-12, CA 15-3 and to estimate the number of copies of CAN SINEs sequences relating them with clinicopathological parameters and survival of the group, in order to infer a likely value as prognostic markers in breast cancer. Through enzyme immunoassay and real-time PCR were evaluated 33 dogs with mammary cancer and 50 control dogs and their concentrations were statistically related with these parameters. Significant difference was found between IL-8 serum levels in bitches with mammary neoplasia in relation to the control group and in relation to disease progression, the lymph node involvement and in relation to death. Low levels of IL-12 were observed in female dogs with tumors with a longer evolution, aged 10 years and in addition, the high survival rate was correlated with high levels of IL-12. Thus, increased IL- 8 and decreased IL-12 can be considered independent prognostic markers which confirm their role as tumorigenic and anti-tumorigenic cytokines, respectively. For the CA 15-3 were found higher levels in dogs with lymph node involvement and poor survival of the group. In estimating the number of copies of CANSINEs, significant difference was found regarding the parameters indicators of poor prognosis and low survival. Our results... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:26:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-08Bitstream added on 2014-06-13T20:54:03Z : No. of bitstreams: 1 silva_cl_me_sjrp.pdf: 1181592 bytes, checksum: 04dbaddabd010ef67a9783ca97045cdc (MD5) / Os tumores mamários da espécie canina prestam-se como modelos apropriados, e válidos, ao estudo da biologia do câncer. O estudo de marcadores prognósticos e preditivos no câncer tem se mostrado efetivo na pesquisa e rotina diagnóstica. Dentre esses marcadores, destacam-se os envolvidos no estresse oxidativo e alterações na capacidade antioxidante celular, que desempenham papel fundamental na patogênese do câncer. A glutationa (GSH) é um dos agentes mais importantes do sistema de defesa antioxidante da célula, pois em conjugação com as enzimas glutationa peroxidase (GSH-Px) e glutationa S transferase pi (GSTpi) desempenha um papel central na biotransformação e detoxificação de drogas quimioterápicas. O objetivo deste estudo foi avaliar a expressão da proteína GSH e das enzimas GSTpi e GSH-Px por imunohistoquímica em tumores de mama em cadelas, a fim de inferir o seu valor prognóstico, e validar por análise molecular, verificando a resistência das células neoplásicas ao quimioterápico, bem como o nível de estresse oxidativo avaliado bioquimicamente pelo malondialdeído (MDA), e a avaliação da atividade antioxidante total em equivalência ao Trolox (TEAC) . As proteínas foram detectadas no tecido tumoral de 30 cadelas por imunohistoquímica e quantificadas pela técnica de densitometria óptica. As médias relacionadas à expressão das proteínas foram estatisticamente comparadas com as características clínico-patológicas das cadelas. A expressão dos genes glutamato cisteina ligase (GCLC) e glutationa sintetase (GSS) que sintetizam a GSH e do gene GSH-Px foi verificada por qPCR em células neoplásicas de mama cultivadas e submetidas ao tratamento com doxorrubicina. A dosagem sérica do MDA e TEAC foi realizada em cadelas no momento da cirurgia, um, seis e 18 meses após... / The mammary tumors of canine species lend themselves as models appropriate, and valid, to study of cancer biology. The study of prognostic and predictive markers in cancer have been proven effective in research and diagnostic routine. Among these markers include those involved in oxidative stress and changes in antioxidant capacity, which play a fundamental role in the pathogenesis of cancer. Glutathione (GSH) is one of the most important agents of the antioxidant defense system of the cell because in conjunction with glutathione peroxidase (GSH-Px) and glutathione S transferase pi (GSTpi) plays a central role in the biotransformation and detoxification of drugs chemotherapy. The objective of this study was to evaluate the protein expression of GSH and the enzymes GSH-Px and GSTpi by immunohistochemistry in breast tumors of dogs in order to infer its prognostic value, and prospective study to evaluate them by molecular analysis, verifying the resistance of cancer cells to chemotherapy, as well as the level of oxidative stress assessed biochemically by malondealdehyde (MDA), and the evaluating the total antioxidant activity equivalent to Trolox (TEAC). The proteins were detected in tumor tissue of 30 female dogs by immunohistochemistry and quantified by optical densitometry technique. The mean expression of related proteins were statistically compared with the clinical- pathologic of the bitches. The expression of genes glutamate cysteine ligase (GCLC) and glutathione synthetase (GSS) that synthesize GSH and of GSH-Px gene was verified by qPCR in breast cancer cells grown and subjected to treatment with doxorubicin. The serum dosage of MDA and TEAC was performed in betches at the time of surgery, one, 6 and 18 months after surgery and compared with dogs in the control group. The results showed... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:33:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-07-15Bitstream added on 2014-06-13T19:04:13Z : No. of bitstreams: 1 silveira_act_dr_jabo.pdf: 1179279 bytes, checksum: 7f06f3c7d496e3a8bb5d2796f59b3bd6 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Tendo em vista a elevada incidência das neoplasias mamárias nas cadelas, este trabalho teve como objetivo avaliar as células inflamatórias infiltradas e também a presença de citocinas pró e anti-inflamatórias nos tumores malignos bem como investigar os marcadores de prognóstico nas mesmas neoplasias. Para a realização deste estudo foram selecionadas 30 amostras de tumores mamários de cadelas. As amostras foram divididas em 3 grupos, de acordo com a classificação histopatológica sendo, carcinossarcoma, carcinoma sólido, carcinoma tubulopapilar. A avaliação das células inflamatórias CD3 (Dako), CD4 (Novocastra), CD8 (Novocastra), CD79-a (Dako), CD68 (Dako), citocinas TGF-b (Santa Cruz), IL-2r (Santa Cruz) e dos marcadores de prognóstico Ki-67, p53, ER e PR, foi conduzida por imuno-histoquímica, utilizando-se a técnica estreptoavidina-biotina-peroxidase. Em relação aos resultados obtidos, observou-se maior concentração de linócitos B nos carcinomas sólidos e tubulopapilares, o que também foi visto para a citocina TGF- podendo-se sugerir que essa citocina esteja auxiliando a resposta imune humoral do hospedeiro favorecendo assim o tumor. Esses dois marcadores apresentaram diferença estatística significativa (P<0,05) entre a imunorreatividade do carcinossarcoma e os outros dois tumores. Houve diferença significativa (P<0,05) entre a imunorreatividade dos três tipos de tumores para o marcador ER. Para o PR, a diferença significativa foi vista entre o carcinoma sólido e carcinoma tubulopapilar, o mesmo sendo visto para o Ki-67. O gene p53 apresentou diferença significativa (P<0,05) entre a imunorretaividade do carcinossarcoma e do carcinoma sólido e tubulopapilar, o mesmo acontecendo para o MHC II. Não houve correlação entre a resposta inflamatória e os marcadores de prognóstico. Houve correlação negativa entre o p53 e os receptores de progesterona e estrógeno. / In view of the high incidence of mammary neoplasia in dogs, the aim of this study was to evaluate the inflammatory infiltrate cells and presence of pro and antiinflammatory cytokines in malignant tumors, and to investigate the prognostic markers in these neoplasms. For this, a sample of 30 mammary tumors from dogs was selected. The samples were divided into three groups according to histopathological classification: carcinosarcoma, solid carcinoma or tubulopapillary carcinoma. The CD3 (Dako), CD4 (Novocastra), CD8 (Novocastra), CD79-a (Dako) and CD68 (Dako) inflammatory cells, TGF-b (Santa Cruz) and IL-2r (Santa Cruz) cytokines and Ki-67, p53, estrogen receptor and progesterone receptor prognostic markers were evaluated by means of immunohistochemistry, using the streptavidin-biotin-peroxidase technique. It was found that the B lymphocyte concentration was greater in solid and tubulopapillary carcinomas. This was also seen for TGF-b cytokines, which may suggest that this cytokine was helping the host’s humoral immune response, thereby favoring the tumor. The immunoreactivity of these two markers was significantly different (P<0.05) between carcinosarcomas and the other two tumor types. There were significant differences (P<0.05) in immunoreactivity between the three tumor types for the estrogen receptor marker. For progesterone receptors and Ki-67, a significant difference was seen between solid and tubulopapillary carcinomas. For the gene p53 and MHC II, there was a significant difference (P<0.05) in immunoreactivity between carcinosarcomas and the other two tumor types. There was no correlation between the inflammatory response and the prognostic markers. There was a negative correlation between p53 and the progesterone and estrogen receptors.