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Avaliação imunoistoquimica da cérvix de cadelas saudáveis e com o diagnóstico de piometra / Cervix immunohistochemical evaluation of healthy bitches and bitches diagnosed with pyometraVolpato, Rodrigo [UNESP] 21 January 2015 (has links) (PDF)
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000857897.pdf: 1190502 bytes, checksum: 015b214298e4307571cdcfa119179945 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo desse estudo foi avaliar os mecanismos envolvidos na abertura cervical em cadelas saudáveis e com o diagnóstico piometra. Para tanto, foram realizadas avaliações imunoistoquímicas em diferentes regiões da cérvix: epitélio luminal, região glandular e região muscular, para os receptores e-NOS, i-NOS, FAS, FAS-L, PGF2α-R e COX-2. Foram utilizadas 11 cadelas com piometra aberta, 11 cadelas com piometra fechada, 6 cadelas saudáveis em fase folicular e 6 em fase luteal do ciclo estral. Todas as fêmeas foram submetidas a ovariohisterectomia (OSH) e dosagens de progesterona foram realizadas para confirmação da fase do ciclo estral. As dosagens foram realizadas utilizando-se kits comerciais em fase sólida. Para avaliar a imunomarcação foram utilizados dois métodos. No primeiro, em cada um desses tecidos foram avaliados cinco campos, cada campo recebeu um escore de 0 a 4 (sendo 0 = 0 a 5% do campo positivo, 1 = 5 a 25% do campo positivo, 2 = 25 a 50%, 3 = 50 a 75% do campo positivo e 4 = 75 a 100% do campo positivo) sendo considerados positivos aqueles corados em marrom e negativos aqueles corados em azul. No segundo método a intensidade das marcações foi avaliada de forma subjetiva em fraca, moderada e forte, recebendo respectivamente escore 1, 2 e 3 para a análise estatística. As leitura foram realizadas em cinco campos aleatórios, sendo todos os campos homogêneos (mais de 80% da coloração com a mesma intensidade). As avaliações foram realizadas em um microscópio de luz no aumento 400 vezes. Foram realizados os testes estatísticos não-paramétrico de Mann-Whitney para comparar a presença, a concentração de cada receptor e a região entre os grupos piometra fechada e aberta. Entre os grupos piometra, fase luteal e fase folicular, utilizou-se o teste Kruskall-Wallis, pois a comparação foi realizada entre 3 grupos independentes. Os receptores i-NOS e FAS comportam-se da mesma maneira em cadelas com piometra... / The aim of this study was to evaluate the mechanisms involved in cervical opening in bitches with pyometra and healthy bitches. To this end, immunohistochemical evaluations were conducted in different regions of the cervix, epithelium luminal, glandular and musculene for e-NOS, i-NOS, FAS, FAS-L, PGF2α-R and COX-2 receptors. For this purpose, were used 11 bitches with open pyometra, 11 bitches with close pyometra, 6 healthy bitches on follicular phase and 6 on luteal phase of the estrous cycle. All females were subjected to ovariohysterectomy (OSH) and serum progesterone were performed to confirm the phase of the estrous cycle. The measurements were performed using solid phase commercial kits. For immunostaining evaluation two methods were used. In the first, five fields were evaluated in each of these tissues, where each field received a score from 0 to 4 (0 = 0 to 5% of positive field, 1 = 5 to 25% of positive field, 2 = 25 to 50%, 3 = 50 to 75% of positive field and 4 = 75 to 100% of positive field) being considered positive those stained in brown and blue the negative ones. In the second method the markings intensity was assessed subjectively in weak, moderate and strong, respectively receiving scores 1, 2 and 3 for statistical analysis. Readings were taken at five random fields, that were all homogeneous (more than 80% of the staining with the same intensity). The evaluations were performed using a light microscope at a magnification of 400x. Non-parametric statistical test of Mann-Whitney was performed to compare the presence, concentration of each receiver and the region between the closed and open pyometra groups. Among pyometra, follicular phase and luteal phase groups, Kruskall-Wallis test was used, once the comparison made was between three independent groups. The i-NOS and FAS receptors behave in the same way in bitches with open and closed pyometra, not being involved in cervical opening mechanism in this disease. The ...
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Intestinal responses to Clostridium perfringens in broilersRussell, Katherine Margaret January 2016 (has links)
Clostridium perfringens is the aetiological agent of Necrotic enteritis (NE); a disease that impacts on the health and welfare of broilers. This disease is a large cost to the industry and presents as lesions in the small intestine hindering productivity. Antibiotics are commonly used to treat NE but as pressure increases to limit their use further information about disease onset and broiler responses to the bacteria and it’s virulence factors during infection is required to implement new preventative measures and treatments. NetB is a secreted toxin from C. perfringens which has an important role in NE onset. Using an in situ intestinal loop model we have been able to characterise: I) temporal broiler responses to NetB positive bacterial culture supernatant (Chapter 2), ii) early host responses to different isolates possessing NetB (virulent) or not (avirulent) in the presence or absence of bacterial cells (Chapter 3) and iii) the responses of two commercial broiler breeds (Chapter 4) four hours post exposure. Samples collected from these experiments have been used for histology, mRNA expression and immunohistology. We have shown differences in mRNA expression in the duodenum of broilers after exposure to C. perfringens cells as well as the culture supernatant from the isolates used after four hours. The presence of bacteria cells resulted in up-regulation of pro-inflammatory cytokine, IFN-γ, mRNA, whereas it resulted in down-regulation of B-LA, mRNA a gene involved in presentation of pathogens to immune cells. IL-6 mRNA expression was also reduced in the presence of virulent isolates. This could indicate a possible evasion strategy for C. perfringens in broilers. Immunohistochemical analysis indicated that slower growing broilers have increased numbers of immune cells (macrophages and γδ T cells) in their duodenum compared with faster growing broilers, although this did not appear to have an effect on mRNA expression levels of pro-inflammatory cytokines, 4h post antigen infusion. Overall we detect greater changes when bacteria are included with culture supernatant and have highlighted possible mechanisms for C. perfringens to avoid the broiler immune system. Induction of NE in the literature requires pre-disposing factors, including co-infection with other intestinal pathogens and dietary manipulation of the host. The final experiment trialled protocols administering a virulent isolate of C. perfringens in-feed and via gavage along with an increased protein source to induce NE (Chapter 5). These models were not considered to be consistent for further investigation of NE in the future.
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Neurotrophin receptors in select cutaneous malignancies with a propensity for perineural invasionFrydenlund, Noah 08 April 2016 (has links)
Perineural invasion (PNI) in cutaneous squamous cell carcinoma (cSCC) and desmoplastic melanoma (DM) may be a negative prognostic finding, and likely contributes to increased rates of local recurrence. The biological mechanisms underlying PNI remain unclear, although several lines of evidence implicate neurotrophins and their receptors. Expression of the high affinity nerve growth factor (NGF) receptor TrkA has been shown to be associated with PNI in numerous malignancies, although literature in cutaneous neoplasms is sparse. Given this, we sought to ascertain the incidence of PNI in a cohort cSCCs using double immunostaining (DIS), and to investigate PNI's relationship with TrkA expression and established histopathologic prognosticators. In DMs we investigated the relationship between TrkA and PNI. In DM we additionally analyzed expression of the low affinity NGF receptor (p75NGFR) and the presence of a functional polymorphism in the glial cell line-derived neurotrophic factor (GDNF) receptor RET (RETp) as they relate to PNI.
In this IRB approved study, cSCCs from the head and neck (H&N) and 53 from non-H&N areas were immunohistochemically analyzed for PNI (DIS with S100 and p63) and TrkA expression. For DM, 43 cases were immunohistochemically evaluated for TrkA and p75NGFR expression while RETp was detected by direct DNA sequencing. The presence of each was correlated with histologically observed PNI.
In cSCCs, comparing H&N versus non-H&N areas; using hemotoxylin and eosin (H&E) PNI was detected in 11% versus 6% of cases respectively and using DIS, in 23% versus 15% respectively, with significant disagreement between both methods (𝜅=0.47, p=0.002). There was a 2.33 fold increase in PNI detection with DIS compared to H&E (95%CI: 1.12-4.87; p=0.02). TrkA expression was 2.9 times more frequently observed in cSCCs from the H&N compared to those from non-H&N areas (p=0.01). Regardless of site, TrkA expression was associated with decreased degree of differentiation (OR=6.46, p=0.0006) and high-risk morphologic variants (OR = 6.53, p=0.002). TrkA expression was not significantly associated with PNI (p=0.33).
In DM, PNI was present in 67% of cases. On univariate analysis; p75NGFR was associated with PNI (expression detected in 79% of PNI-positive cases compared to 36% of PNI-negative cases, p=0.005), increased Breslow's depth and greater Clark's Level (p= 0.007 and p= 0.01 respectively). RETp was noted in 28% of cases but was not significantly associated with PNI (p=0.27) or other histopathologic variables. TrkA expression was absent in all cases. PNI was associated with increased Breslow's depth and Clark's Level (p=0.01 and p=0.009 respectively). Controlling for the association between p75NGFR and depth, p75NGFR remained associated with an increased propensity for PNI (OR=4.68, p=0.04).
In conclusion, increased PNI detection with DIS in cSCCs underscores the adjunctive utility of immunohistochemistry in microstaging. Although unlikely to play a role in the development of PNI, TrkA's association with cSCCs from H&N and select histopathologic parameters suggests a role for the NGF-TrKA axis in tumorogenesis while its absent expression in DM suggests that expression is lineage-related. Lastly, In DM, p75NGFR expression is significantly associated with PNI and a more locally aggressive phenotype.
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Validação de anticorpo monoclonal anti-CD45 obtido in house para utilização em citometria de fluxo e imuno-histoquímicaSodré, Luciandro Pereira [UNESP] 27 February 2009 (has links) (PDF)
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sodre_lp_me_botfm.pdf: 1339623 bytes, checksum: ed335773fb6c0b581fc33a31e3612a6e (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Anticorpos monoclonais (AcMm) são imunoglobulinas produzidas por engenharia celular através da fusão de células tumorais com linfócitos B provenientes de organismos previamente imunizados contra o antígeno de interesse. As células recém criadas recebem a denominação de hibridomas, crescem indefinidamente em cultura por agregar características de imortalidade das células tumorais. A tecnologia de monoclonais compreende etapas seriadas, incluindo imunização, fusão, screening, clonagem, caracterização de especificidade dentre outras. O CD45 é um marcador expresso na superfície de todos os leucócitos humanos: linfócitos, eosinófilos, monócitos, basófilos e neutrófilos sendo, o maior componente da membrana de linfócitos, o que aumenta a probabilidade de obtenção de AcMm com este perfil em protocolos de imunização com linfócitos. Este marcador é amplamente utilizado em diagnóstico por citometria de fluxo, no acompanhamento de pacientes HIV+, na classificação de leucemias e linfomas, quantificação de células CD34+ para transplantes de medula óssea e, também em imunohistoqu ímica auxiliando na subtipagem das neoplasias, principalmente linfomas (Hodgkin, não Hodgkin e outras categorias) e leucemias de células B e T. Assim, este trabalho teve por objetivo a caracterização de anticorpo monoclonal anti-CD45 desenvolvido in house e validação deste em aplicações diagnósticas envolvendo citometria de fluxo e imunohistoqu ímica, comparando-o com anticorpos monoclonais comerciais, visando minimização dos custos dessas técnicas. Foram utilizadas 50 amostras de sangue excedente de doadores para o grupo controle e 48 amostras de sangue de pacientes HIV+ para validação do AcMm em citometria de fluxo e analisados 114 fragmentos em lâmina de Tissue Microarray - TMA provenientes de 38 amostras de amígdalas para validação do AcMm em imunohistoqu ímica... / Monoclonal antibodies (mAb) are immunoglobulins produced by cell engineering through fusion of tumor cells and B lymphocytes from organisms previously immunized against the antigen of interest, generating thus hybridomas that indefinitely grow in culture since they aggregate immortality characteristics of tumor cells capable of secreting antibodies. The production of such antibodies consists of sequential steps, including immunization, fusion, screening, cloning, and specificity characterization. CD45 is a marker expressed in the surface of all human leukocytes: lymphocytes, eosinophils, monocytes, basophils, and neutrophils. Besides, CD45 is the major component of lymphocyte membranes, which increases the probability of obtaining mAb presenting such profile in immunization procedures using lymphocytes. This marker has been widely used in diagnosis through flow cytometry, besides monitoring of HIV+ patients, classification of leukemia and lymphomas, quantification of CD34+ cells for bone marrow transplantation, and also in immunohistochemistry, helping in subtyping of neoplasms, mainly lymphomas (Hodgkin s, non-Hodgkin and other categories) and B- and T-cell leukemia. Thus, this work aimed to characterize the in-house-developed anti-CD45 monoclonal antibody besides its validation in diagnostic applications involving flow cytometry and immunohistochemistry through comparison with commercial monoclonal antibodies in order to reduce the costs of such techniques. Fifty samples of extra blood from donors were used in the control group, and 48 blood samples from HIV+ patients were used for mAb validation in flow cytometry. For mAb immunohistochemical validation, 114 fragments from 38 amygdala samples were analyzed in a Tissue Microarray - TMA slide. In flow cytometry, the commercial anti-CD45 and the likely in-house-obtained anti-CD45 (LINB5 clone) did not significantly differ concerning cell... (Complete abstract click electronic access below)
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Avaliação histoquímica do colágeno e expressão imunoistoquímica de metaloproteinases e alfa-actina de músculo liso no sarcoide equino / Histochemical evaluation of collagen and immunohistochemical expression of metalloproteinases and alpha-smooth muscle actin in equine sarcoidScarelli, Sarah Paschoal [UNESP] 27 October 2015 (has links) (PDF)
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000866499.pdf: 5832754 bytes, checksum: 88ad83a6f6d6d0a52fd9b7ed2380f8f6 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O sarcoide equino é a neoplasia de pele mais prevalente na espécie equina, possuindo seis tipos clínicos descritos: verrucoso, misto, maligno, nodular, oculto, fibroblástico e o misto. Evidências demonstram uma importante influência do estroma no desenvolvimento e progressão dos tumores malignos. O arranjo e a quantidade de colágeno, a presença de miofibroblastos no estroma ou como componentes da neoplasia, assim como a expressão de metaloproteinases (MMPs) pelas células tumorais, tem sido apontados como elementos que podem indicar o comportamento da neoplasia. Os objetivos do presente trabalho foram associar aos diferentes tipos clínicos de sarcoide equino as expressões de MMP-1, MMP-2 e MMP-9, identificar a presença ou não de miofibroblastos pela expressão de α-SMA e as características e distribuição do colágeno presentes, afim de determinar possíveis marcadores do comportamento biológico da neoplasia. As características morfológicas microscópicas típicas dos sarcoides equinos estavam presentes nas amostra avaliadas, mas estas por si só não podem caracterizar um tipo clínico específico. O estroma tumoral nos tipos clínicos fibroblástico e verrucoso, nos cortes corados pelo método de picrosirius red e examinados sob luz polarizada, era constituído por fibras colágenas frouxas e delicadas, compatível com fibras imaturas comuns em lesões menos estáveis. Os miofibroblastos foram identificados no sarcoide equino e a variação observada na sua distribuição e intensidade de expressão de alfa-SMA foi observada mesmo dentro do mesmo tipo clínico. Os fibroblastos neoplásicos dos sarcoides equinos demonstram expressão MMP-1, MMP-2 e MMP-9. Os tipos clínicos verrucoso e misto apresentam maior expressão de MMP-2 do que tipo clínico nodular, sendo este achado compatível com o potencial de agressividade maior. Pode-se verificar que existem diferenças entre os tipos clínicos de sarcoide equino / The equine sarcoid is the most prevalent skin cancer in equine, having six clinical types described: warty, mixed, malignant, nodular, hidden, fibroblast and mixed. Evidence has shown an important influence on stromal development and progression of malignant tumors. The arrangement and quantity of collagen, the presence of myofibroblasts in the stroma or as a tumor component as well as the expression of matrix metalloproteinases (MMPs) by tumor cells, have been identified as factors that may indicate the behavior of the tumor. The objectives of this study were associate the different clinical types of equine sarcoid to expressions of MMP-1, MMP-2 and MMP-9, to the presence or absence of myofibroblasts by α-SMA expression and the characteristics and distribution of collagen, in order to determine possible markers of the biological behavior of the tumor. Typical microscopic morphological characteristics of equine sarcoid were present in the samples evaluated, but these alone can't characterize a especific clinical type. In sections stained with picrosirius red method and examined under polarized light the tumor stroma in fibroblastic and verrucous clinical types, was composed of loose and delicate collagen fibers, compatible with immature fibers in less stable lesions. Myofibroblasts were identified in equine sarcoid and the variation in distribution and intensity of alpha-SMA expression was observed even within the same clinical type. The neoplastic fibroblasts in equine sarcoid demonstrate expression MMP-1, MMP-2 and MMP-9. The verrucous and mixed types have higher MMP-2 expression than nodular clinical type, and this finding is consistent with the more aggressive potential. So there are differences between the clinical types of equine sarcoid that can be seen by the use of histochemical and immunohistochemical techniques
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Expression of regulatory t cells in polar leprosy and leprosy reactions / Estudo da expressÃo imunohistoquÃmica de linfÃcitos T-REG em pacientes com formas polares de hansenÃase e reaÃÃes hansÃnicasDaniela Ventin Prates Rezende 16 July 2013 (has links)
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Leprosy is a chronic infectious disease caused by Mycobacterium leprae and presents a wide range of clinical manifestations, depending on the host immune response. At the tuberculoid pole, patients have an effective Th1 cellular immune response against bacillus, with well-demarcated granulomas, which limits the disease, while the lepromatous pole is characterized by the absence of specific cellular immunity against Mycobacterium leprae, with a Th2 immune response, uncontrolled proliferation of leprosy bacilli and a spread disease. In addition to Th1 and Th2 lymphocytes, recently other subset of T cells has been described. T regulatory lymphocytes play an immunosuppressive role in the activity of other cells and seem to participate in modulating the immune response in human chronic infections and self-tolerance. We performed immunohistochemical study on skin biopsies of 35 patients with different forms of leprosy using anti-CD4, anti-CD8 and anti-FoxP3 antibodies. There was an increased CD4 expression in tuberculoid patients and an increased CD8 expression in lepromatous patients. The FoxP3 expression was higher in lepromatous and reversal leprosy reaction patients. As previously demonstrated in other granulomatous infections, T regulatory cells may play a role in the immunopathology of leprosy. However, studies with larger series of patients are required to further elucidate the relationship between the presence of these cells and the clinical form of the disease. / A hansenÃase à uma doenÃa infecciosa crÃnica causada pelo Mycobacterium leprae e apresenta uma grande variedade de manifestaÃÃes clÃnicas que dependem da resposta imunolÃgica do hospedeiro. Em um polo està a forma tuberculoide, na qual os pacientes apresentam uma resposta imunolÃgica Th1 celular contra o bacilo, com formaÃÃo de granulomas bem delimitados com doenÃa localizada. No outro polo temos a forma virchowiana caracterizada pela ausÃncia de imunidade celular especÃfica ao Mycobacterium leprae, com padrÃo de resposta imunolÃgica Th2, com macrÃfagos repletos de bacilos, mas com poucos linfÃcitos, sem formaÃÃo de granulomas e com disseminaÃÃo da doenÃa. AlÃm dos linfÃcitos Th1 e Th2, outra populaÃÃo de linfÃcitos T foi descrita recentemente. Os linfÃcitos T regulatÃrios desempenham um papel imunossupressor na atividade de outros linfÃcitos e de outras cÃlulas inflamatÃrias e parecem participar da modulaÃÃo da resposta imunolÃgica em infecÃÃes crÃnicas em seres humanos e da autotolerÃncia. Neste trabalho, foram analisadas 35 biÃpsias cutÃneas de lesÃo de pacientes portadores de diferentes formas de hansenÃase quanto à expressÃo dos marcadores celulares CD4, CD8 e FoxP3 por imunohistoquÃmica. Enquanto a expressÃo do CD4 foi maior nos pacientes portadores da forma tuberculoide da hansenÃase, a expressÃo do CD8 mostrou-se maior nos portadores de hansenÃase virchowiana. Quanto à expressÃo do FoxP3, esta foi maior nos portadores da forma virchowiana da hansenÃase e da reaÃÃo hansÃnica tipo 1 em relaÃÃo aos indivÃduos sadios. Parece que, assim como jà demonstrado em outras doenÃas infecciosas granulomatosas, os linfÃcitos Treg participam da imunopatologia da hansenÃase. Entretanto, estudos com maior nÃmero de pacientes sÃo necessÃrios, para um melhor entendimento da relaÃÃo entre a presenÃa desses linfÃcitos e a forma clÃnica da doenÃa.
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Analise da expressão de KI-67, P53, MCM 2 e MCM 5 em leucoplasia verrucosa proliferativa. / Analysis of KI-67, P53, MCM 2 e MCM 5 expression in oral proliferative verrucous leukoplakiaGouvêa, Adriele Ferreira 27 February 2008 (has links)
Orientador: Marcio Ajudarte Lopes / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-10T15:16:37Z (GMT). No. of bitstreams: 1
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Previous issue date: 2008 / Resumo: Leucoplasia verrucosa proliferativa é uma forma agressiva de leucoplasia oral, que afeta mais mulheres acima de 60 anos, sem história de etilismo e tabagismo. Tem apresentação clínica multifocal, mostra recorrência e sempre desenvolve displasia epitelial e carcinoma espinocelular (CEC). O objetivo deste trabalho foi avaliar os achados clínico-patológicos e a expressão imunohistoquímica de ki-67, p53, Mcm2 e Mcm5, na tentativa de elucidar o comportamento biológico distinto desta condição. Os dados clínico-patológicos de 12 pacientes foram revisados. Todos eram mulheres, acima de 50 anos (50% acima de 70 anos), 91,7% eram não fumantes e nenhuma etilista habitual. Os casos acompanhados por mais tempo mostraram aspectos clínicos compatíveis com lesões iniciais com presença de hiperqueratose e acantose, desenvolvimento de diferentes graus de displasia e CEC. Os locais mais acometidos foram rebordo alveolar (66,6%), língua (50%) e mucosa jugal (41,6%). Quatro pacientes desenvolveram CEC. Os achados imunohistoquímicos mostraram expressões variáveis em cada marcador. As áreas epiteliais sem displasia mostraram imunopositividade fraca para p53 e ki- 67 e moderada para Mcm2 e Mcm5; os casos de displasia leve tiveram expressão fraca de p53, moderada de ki-67 e Mcm5 e forte de Mcm2; displasia moderada exibiu marcação fraca de Mcm5 e de p53 e Mcm2, além de expressão moderada de ki-67; CEC mostrou expressão fraca de Mcm5, moderada de Mcm2 e forte de ki-67 e p53. Não foi observado, na maioria dos casos, um padrão regular de aumento de marcação imunohistoquímica conforme avanço do grau de displasia epitelial. A expressão de ki-67, Mcm2 e Mcm5 mostrou-se variada, com casos de displasias leves apresentando forte imunopositividade / Abstract: Proliferative verrucous leukoplakia (PVL) is an aggressive form of oral leukoplakia that affects particularly women over 60 years of age, without tobacco and alcohol intake history. Has multifocal presentation, presents recurrences and always develops epithelial dysplasia and squamous cell carcinoma (SCC). The aim of this study was to evaluate the clinico-pathological findings and the ki-67, p53, Mcm2 and Mcm5 immunohistochemical expression, in order to elucidate the distinct biologic behavior of this condition. The clinico-pathological data of 12 patients were reviewed. All were women, above 50 years (50% above 70 years of age), 91.7% were non smoker and 100% were non-drinker. The cases with longer follow-up showed clinical aspects compatible with initial lesions presenting hyperkeratosis and acantosis, development of different grade of epithelial dysplasia and SCC. Alveolar ridge (66,6%), tongue (50%) and buccal mucosa (41,6%) were the most affected sites. Four patients developed SCC. The immunohistochemical findings showed variable expression for each antibody. Areas without epithelial dysplasia showed weak positivity to p53 and ki-67 and moderate to Mcm2 and Mcm5. Mild dysplasia cases showed weak positivity to p53, moderate to ki-67and Mcm5 and strong to Mcm2; moderate dysplasia exhibited weak p53, Mcm2 and Mcm5 expression and moderated ki-67 imunopositivity; SCC showed weak Mcm5 positivity, moderate expression of Mcm2 and strong imunopositivity of ki-67 and p53.The majority of the cases did not show a regular pattern of increasing immunohistochemical expression according to higher grades of epithelial dysplasia. Ki-67, Mcm2 and Mcm5 showed variable imunopositivity, with some mild dysplasia cases presenting strong expression / Mestrado / Patologia / Mestre em Estomatopatologia
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Expressão das proteinas da familia PLUNC nas glandulas salivares maiores de pacientes autopsiados com AIDS em fase avançada e sem AIDS / Expression of PLUNC family protein in major salivary gland of the autopsied patients with advanced AIDS and without AIDSSilva, Andreia Aparecida da 13 August 2018 (has links)
Orientadores: Pablo Agustin Vargas, Lynne Bingle / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-13T01:09:27Z (GMT). No. of bitstreams: 1
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Previous issue date: 2009 / Resumo: Introdução: Inúmeras lesões de origem infecciosa, cística, neoplásica e inflamatória foram reportadas nas glândulas salivares de pacientes HIV+. Objetivos: Os objetivos deste trabalho foram analisar e comparar a resposta do sistema imune inato (proteínas da família PLUNC) em glândulas salivares maiores, provenientes de pacientes autopsiados com AIDS e sem AIDS (grupo controle) no Departamento de Patologia da Faculdade de Medicina da Universidade Estadual de São Paulo (FMUSP) no período de 1996 a 2000. Material e Métodos: Os pacientes autopsiados foram divididos em 05 grupos: grupo 01- controle (pacientes HIV negativos), grupo 02- HIV+ sem alterações nas glândulas salivares maiores, grupo 03- (micobacteriose), grupo 04 (citomegalovirose) e grupo 05 (sialadenite) para a realização de reações de imunohistoquímica para os anticorpos SPLUNC 1, SPLUNC 2 A, SPLUNC 2B e LPLUNC 1. Para o grupo controle foi realizado técnica de hibridização in situ para SPLUNC 2. Resultados: a média de idade dos pacientes selecionados para o grupo controle foi de 60,92 anos + 9,48 anos enquanto que a média de idade dos pacientes HIV positivos foi de 37,75 anos + 11,11. Nos casos de micobacteriose e citomegalovirose foi observada maior intensidade de marcação nas regiões próxima a área de infecção, quando comparada com áreas na periferia da lesão para os anticorpos SPLUNC 2 A e 2B. O anticorpo LPLUNC 1 foi positivo apenas nos ductos salivares e apresentou positividade em 42,22%, 51,06% e 63,88% para as glândulas parótida, submandibular e sublingual respectivamente. Com relação à hibridização in situ, foi observado positividade em todos os casos. Conclusão: a família de proteínas PLUNC pode ter papel fundamental na proteção dos organismos frente a agentes infecciosos, no entanto são necessários maiores estudos. / Abstract: Objectives: The aim of this study was to determine the expression of PLUNC proteins in major salivary glands (MSG) of AIDS patients with or without infectious conditions and non-HIV patients using post-mortem material. Methods: Sex, age, CD4 cell count, and clinical history were obtained retrospectively from the clinical records of all patients (n=63). We analysed the expression of PLUNCs (SPLUNC1, SPLUNC2, LPLUNC1) using immunohistochemistry in parotid (n=45), submandibular (n=47) and sublingual gland (n=37) samples of AIDS patients [30 with normal histology, 21 with mycobacteriosis, 14 with cytomegalovirus (CMV) infection, 30 with chronic nonspecific sialadenitis, and 30 HIV-negative controls. In situ hybridization (ISH) for SPLUNC2 in the MSG of the HIVnegative group was performed. Immunoreactivity was assessed as positive or negative. Results: The mean age of the patients who died of AIDS (n=63) and CD4 cell count (n=44) were 37 years and 63 cells microL(-1), respectively. The mean age of the HIV negative patients was 61 years. SPLUNC 1 expression was detected in the mucous acini of submandibular and sublingual glands, and SPLUNC2 was seen in the serous cells of the MSG. LPLUNC 1 expression was only positive in the salivary ducts of the MSG. There was a higher expression of SPLUNC2 in AIDS patients with CMV infection and mycobacteriosis when compared with all other groups. The intensity of staining for SPLUNC2 was greater around the lesions than the peripheral ones. There were no significant differences between control subjects and AIDS patients without histologic alterations or with chronic nonspecific sialadenitis. ISH for SPLUNC 2 showed perinuclear positivity in the serous cells in all HIV-negative cases. Conclusions: SPLUNC1 and LPLUNC1 proteins were similarly expressed in the MSG of AIDS patients and non-HIV patients. CMV infection and mycobacteriosis increase SPLUNC2 expression in serous cells in the MSG of AIDS patients. Further studies are needed to understand the biological processes involved in SPLUNC2 expression in the MSG infected by CMV and mycobacteriosis. / Doutorado / Patologia / Doutor em Estomatopatologia
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Expressão da proteina erbB-2 e dos receptores de esteroides na transição das frações in situ e invasora de neoplasias de mama / erbB-2 expression and hormone receptor status in the transition areas from in situ to invasive ductal carcinoma of the breastPeres, Raquel Mary Rodrigues, 1983- 13 August 2018 (has links)
Orientador: Luis Otavio Sarian / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-13T16:46:52Z (GMT). No. of bitstreams: 1
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Previous issue date: 2009 / Resumo: Introdução: Há evidências indicando que a transição de carcinoma ductal in situ (CDIS) para carcinoma ductal invasivo (CDI) é regulada por um número restrito de genes. Ainda permanece obscuro se a expressão do receptor erbB-2 e dos receptores hormonais predizem o potencial invasivo dos CDIS. Objetivos: Avaliar a expressão dos receptores de estrógeno e progesterona (RE/RP) e da proteína erbB-2 em áreas específicas em tumores que apresentem componentes de CDIS e CDI concomitantemente, indicando uma possível transição. Materiais e Métodos: Foram selecionados 85 casos de carcinomas da mama apresentando regiões contíguas de CDIS e CDI e obtidas amostras histológicas separadas, uma da região CDIS e outra da região CDI, do bloco de parafina original do diagnóstico, para montagem de um TMA (tissue microarray). A expressão das proteínas erbB-2, RE e RP foram verificadas através da técnica convencional de imuno-histoquímica. Os dados clínicos das pacientes foram coletados por consulta ao prontuário. Resultados: Aproximadamente 35% das áreas invasivas foram classificadas como 2+ ou 3+ para o erbB-2, em contraste a 47% no componente invasivo correspondente. A expressão de erbB-2 não se mostrou diferente nos componentes in situ e invasivo (p=0,12). A expressão de RP não foi estatisticamente diferente comparando-se regiões in situ e invasivas do tumor (p=0,06). Em contraste, houve uma discreta mudança na expressão de RE (p=0,04). O coeficiente de concordância intraclasse (ICC) revelou forte concordância nas comparações entre as amostras de erbB-2 (ICC=0,61), RP (ICC=0,61) e RE (ICC=0,70), nos componentes in situ e invasivo. Conclusão: Os achados sugerem que a expressão de erbB-2, RE e RP não diferem entre os componentes in situ e invasivo do carcinoma da mama, especialmente nas regiões de transição de um componente a outro. Espera-se que as alterações moleculares ocorridas antes da mudança morfológica do tecido persistam na forma invasiva do tumor. A concomitância da atuação de fatores epigenéticos pode explicar porque alguns tumores in situ evoluem para invasivos, enquanto outros tumores in situ com características moleculares semelhantes não apresentam a mesma evolução, suportando a hipótese de uma base multifatorial para a progressão da doença. / Abstract: Introduction: There is evidence indicating that the transition from ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) is regulated by a restrict number of genes. It remains unknown whether erbB-2 and hormone receptor status predict the invasive potential of DCIS. Objectives: To evaluate estrogen/progesterone receptor (ER/PR) and protein erbB-2 status in the specific areas of breast tumors which share both components of DCIS and IDC, concomitantly, indicating a possible transition. Materials and methods: 85 cases of breast malignancies harboring contiguous regions of DCIS and IDC were selected. Separate histological samples, one from the DCIS and other from the neighboring IDC, from the original paraffin block used on diagnosis, were sampled using tissue microarrays (TMA). The erbB-2 and ER/PR status was assessed using conventional immunohistochemistry techniques. Clinicopathological data from the patients and from the disease were retrieved out of the patients' medical records. Results: Approximatelly 35% of the invasive areas were erbB-2 2+ or 3+, compared to 47% of their in situ counterparts. The expression of erbB-2 did not differ in the in situ and in the invasive components of the breast tumors (p=0.12). The PR status was not statistically different comparing the in situ and invasive regions of the tumors (p=0.06). By contrast, there was a slight imbalance in ER status (p=0.04). The intraclass correlation coefficient (ICC) disclosed good agreement in sample-by-sample comparisons of erbB-2 (ICC = 0.61), PR (ICC= 0.61) and ER (ICC=0.70) expression in the in situ and invasive components. Conclusion: Our findings suggest that the expression of erbB-2 and ER/PR does not differ accross the in situ and invasive components of breast malignancies, especially in the transition region from one component to the other. It is expected that the molecular alterations which take place before the morphologic tissue changes persist in the invasive forms of breast tumors. The ocurrance of concomitant epigenetic factors may explain why some in situ tumors develop to an invasive phenotype, while other in situ tumors with similar molecular characteristics do not have the same evolution, supporting the hypothesis for a multifactorial basis for disease progression. / Mestrado / Ciencias Biomedicas / Mestre em Tocoginecologia
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Estudo morfometrico das infiltrações linfocitarias cutaneas em preparados imunoistoquimicos / Morphometric studies of the skin lymphocytic infiltration on immunohistochemical preparationLira, Mariana Montenegro de Melo 08 April 2008 (has links)
Orientador: Jose Vasallo / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-11T18:25:59Z (GMT). No. of bitstreams: 1
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Previous issue date: 2008 / Resumo: A micose fungóide (MF) corresponde a cerca de 50% de todos os linfomas primários da pele. Trata-se de uma neoplasia epidermotrópica constituída por linfócitos atípicos, com fenótipo predominantemente de linfócito T CD4+. O diagnóstico da MF é freqüentemente muito difícil, tanto pela sobreposição com dermatoses benignas, como pela discordância entre achados clínicos e histopatológicos. Embora a certeza de malignidade repouse na observação do comportamento clínico, parâmetros histopatológicos, morfométricos e imunofenotípicos têm sido utilizados para indicar malignidade. Em estudos prévios realizados pelo nosso grupo, não confirmamos os achados de alguns autores, que enfatizam que níveis elevados da relação CD4:CD8 sugerem malignidade. Entretanto, tivemos a oportunidade de comprovar a utilidade da perda da expressão do marcador linfóide T, CD7, no diagnóstico das lesões linfomatosas. Neste último estudo, ficou-nos a impressão de que a ausência de expressão do CD7 era mais evidente em células linfóides mais volumosas e atípicas. Dando continuidade a esta linha de investigação, os objetivos do presente trabalho são os de determinar o papel da avaliação morfométrica em tecidos incluídos em parafina e submetidos à reação imunoistoquímica com CD3 na diferenciação das infiltrações linfocitárias cutâneas e determinar se existe correlação entre o aumento da área nuclear e o percentual de expressão do CD7 nestas lesões. Foram estudados 30 casos de MF e 11 casos de infiltrados linfóides cutâneos benignos. Imagens de 100 células positivas de cada caso foram capturadas e analisadas utilizando o sistema de análise de imagens Imagelab®, sendo mensurados perímetro (P), área (A) e diâmetros (D) nucleares e calculado o índice de contorno nuclear (ICN= P/?A). Houve diferença estatisticamente significante entre as médias de todas as variáveis (p< 0,05), sendo A a variável com maior poder de discriminação entre os grupos (r² = 80,82%). E observou-se uma tendência de correlação entre a perda de expressão de CD7 e o aumento da área nuclear. Ficou, assim, demonstrado que a avaliação de parâmetros nucleares de células previamente identificadas como linfócitos T através da imunocoloração para o CD3 permite a diferenciação morfométrica entre MF e infiltrações linfocitárias benignas. Este método adiciona critérios objetivos para auxiliar no diagnóstico da MF / Abstract: Mycosis fungoides (MF) represents the most common skin lymphoid neoplasm. In initial stages, differential diagnosis of MF with other benign dermal lymphoid infiltrates (BDLI) may be impossible on morphological basis alone. In previous studies conducted by our group, only loss of immunoexpression of CD7 in MF proved to be of diagnostic help, but not the ratio between immunoexpression of CD4 and CD8. In the present study, 30 cases of MF and 11 cases of BDLI were analyzed, in order to compare morphometric parameters, which could be of diagnostic aid. As CD7 is frequently deleted in MF, immunohistochemical detection of T-cells was made using an antibody to CD3. Images of 100 CD3-positive cells per case in both groups were captured and analyzed using a simple computer program (Imagelab®) for nuclear perimeter (P), area (A), diameters (D) and nuclear contour index (NCI). All parameters showed statistically significant higher values for MF. Area was the variable with the strongest discriminating power between the two groups of patients. There was a tendency to a correlation between the loss of expression of CD7 and increase in nuclear area. Even if morphologic evaluation is not accurate to distinguish benign versus malignant dermal lymphoid infiltrates, due to the variability of size and shape of these cells, a more sensitive method promptly shows this difference. Our results suggest that morphometry of CD3-positive lymphoid cells may add valuable information in the differential diagnosis of MF and benign dermatoses / Doutorado / Anatomia Patologica / Doutor em Ciências Médicas
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