Thesis (Ph.D.) -- University of Texas Southwestern Medical Center at Dallas, 2006. / Not embargoed. Vita. Bibliography: 181-228.

Made available in DSpace on 2015-07-22T20:50:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-05-27. Added 1 bitstream(s) on 2015-08-11T03:25:25Z : No. of bitstreams: 1 Publico-00212.pdf: 1245886 bytes, checksum: 4848d0e58992c5a9ff4b387a14c2534a (MD5) / Objetivos: Analisar a expressão das proteínas CD10 e NM23 por estudo imunohistoquímico do tecido do carcinoma colorretal e da mucosa adjacente. Avaliar a relação da expressão dessas proteínas com os aspectos anatomopatológicos da neoplasia, estadiamento clínico, ocorrência de metástases hepáticas e prognóstico dos doentes. Método: Cento e trinta doentes operados por carcinoma colorretal foram analisados. Bloco de tissue microarray foi confeccionado com tecido neoplásico e com a mucosa não neoplásica adjacente. Estudo imuno-histoquímico foi realizado com anticorpos monoclonais NM23 e CD10 no tecido neoplásico e no tecido não neoplásico da mucosa adjacente. A leitura foi realizada por aparelho de escaneamento de lâminas. A imunoexpressão foi avaliada pelo percentual de células coradas e foram obtidos escores de intensidade. Foram considerados como positivos para CD 10 os tumores que expressavam o marcador em mais de 10% das células neoplásicas. Para NM 23 considerou-se dois grupos divididos em fortes expressores (mais de 50%) e expressores fracos (menos de 50%) das células coradas. Os resultados foram relacionados com as características morfológicas e histopatológicas do carcinoma colorretal, estadiamento clínico, presença de metástases hepáticas e com o prognóstico. No estudo estatístico foram utilizados os testes de Mann-Whitney, Kruskal- Wallis e exato de Fisher. A sobrevivência foi avaliada utilizando a curva de Kaplan- Meier, e o desfecho de comparação entre as curvas foi calculado pelo teste de Long rank. Resultados: Ambos os marcadores CD10 e NM23 apresentaram expressão maior no tecido do carcinoma do que na mucosa não neoplásica adjacente (p<0,0001 para ambos). A imunoexpressão tecidual das proteínas NM23 e CD10 não apresentou relação com o grau de diferenciação celular (p=0,57 e p=0,48, respectivamente), invasão vascular (p=0,85 e p=0,67, respectivamente), invasão linfática (p=0,41 e 0,73, respectivamente), infiltração perineural (p=0,46 e p=0,24, respectivamente) e com o estadiamento pela classificação TNM (p=0,19 para ambos). A imunoexpressão de CD10 no tecido do carcinoma colorretal foi maior (p=0,05) nas neoplasias exofíticas do que nos tumores não exofíticos. A expressão das proteínas NM23 e CD10 não apresentou relação com a incidência de metástases linfonodais (p=0,08 e 0,30, respectivamente). A expressão tecidual dos marcadores NM23 e CD10 não se relacionou com a ocorrência de metástases hepáticas (p=0,59 e p=0,31, respectivamente). A sobrevivência livre de doença mostrou relação significante (p=0,01) com a maior intensidade de imunoexpressão da proteína NM23 no tecido do carcinoma colorretal, o mesmo não ocorrendo com a imunoexpressão da proteína CD10 (p=0,18). A sobrevivência global não mostrou relação com as expressões das proteínas NM23 e CD10 (p=0,13 e p=0,24, respectivamente). Conclusões: O tecido neoplásico do carcinoma colorretal expressou mais intensamente as proteínas NM23 e CD10 do que a mucosa não neoplásica adjacente. A imunoexpressão de CD10 no tecido do carcinoma colorretal foi maior (p=0,05) nas neoplasias exofíticas do que nos tumores não exofíticos. A expressão das proteínas NM23 e CD10 não se relacionou com os demais aspectos anatomopatológicos da neoplasia, com a presença de metástase hepática e com o estadiamento do carcinoma colorretal. Os doentes com imunoexpressão aumentada da proteína NM23 apresentaram sobrevivência livre de doença significativamente maior. A intensidade da imunoexpressão tecidual da proteína CD10 não influenciou a sobrevivência livre de doença e a sobrevivência global não se relacionou com a imunoexpressão das proteínas NM23 e CD10. / Aims: To analyze the tissue expression of the proteins CD 10 and NM 23 through the immunohistochemichal study of the colorectal carcinoma and evaluate the expression relation of these proteins with the anatomopathological aspects of the neoplasia, clinical staging, occurrence of hepatic metastasis and patients’ prognostic. Method: One hundred and thirty operated patients of colorectal carcinoma have been analyzed. A block of tissue microarray was produced with the neoplastic mucosa and with the adjacent non-neoplastic mucosa. An immunohistochemichal study was performed with monoclonal antibodies NM23 and CD10 on the neoplastic tissue and non-neoplastic tissue of the adjacent mucosa. The interpretation of the slides was made by a scanner device. The immunoexpression was evaluated by the percentage of colored cells and the obtained intensity scores. The results were related to the morphological and histopathological characteristics of the carcinoma, clinical staging, presence of hepatic metastasis and to the prognostic of the patients. In the statistic study were used the Mann-Whitney test, the Kruskal-Wallis test and Fisher’s exact test. The analysis of survival was conducted with the use of the Kaplan-Meier curve and the comparison conclusion between the curves was calculated through the Longrank test. Results: Both markers CD10 and NM23 presented a higher expression on the carcinoma tissue rather than on the non-neoplastic adjacent mucosa (p<0,0001 for both). The expression of the proteins NM23 and CD10 did not present any relation to the degree of cellular differentiation (p=0,57 and p=0,48, respectively) , vascular invasion (p=0,85 and p=0,67, respectively), lymphatic invasion (p=0,41and 0,73, respectively), perineural infiltration (p=0,46 and p=0,24, respectively) and with the staging by the TNM classification (p=0,19). The immunoexpression of CD10 on the colorectal carcinoma tissue was higher (p=0,15) on the exophytic neoplasias than on the non-exophytic tumors. The expression of the proteins NM23 and CD10 did not present any relation with the incidence of lymphonodal metastasis (p=0,08 and 0,30, respectively). The tissue expression of the markers NM23 and CD10 did not relate to the occurrence of hepatic metastasis (p=0,59 and 0,31 respectively). The disease-free survival disclosed a significant relation (p=0,01) with a higher intensity of immunoexpression of the protein NM23 on the colorectal carcinoma’s tissue. However, the same did not occur with the immunoexpression of the protein CD10 (p=0,18). The global survival did not show any relation with the expression of the proteins NM23 and CD10 (p=0,13 and p=0,24, respectively). Conclusions: The neoplastic tissue of the colorectal carcinoma expresses more intensely the proteins NM23 and CD10 than the adjacent nonneoplastic mucosa. The expression of the proteins NM23 and CD10 does not relate to the presence of lymphonodal metastasis, hepatic metastasis, degree of cellular differentiation, colonic or rectal localization of the neoplasia, presence of vascular and/or lymphatic invasion, presence of neural infiltration and the staging of the colorectal carcinoma. The patients with increased immunoexpression of the protein NM23 presented a disease-free survival significantly higher. The intensity of the tissue immunoexpression of the protein CD10 did not influence the disease-free survival. The global survival does not relate to the immunoexpression of the proteins NM23 and CD10. / TEDE / BV UNIFESP: Teses e dissertações

Allergic asthma is characterized by airway hyperresponsiveness, inflammation, and a cellular infiltrate dominated by eosinophils. Numerous epidemiological studies have related the exacerbation of allergic asthma with an increase in ambient inhalable particulate matter from air pollutants. This is because inhalable particles efficiently deliver airborne allergens deep into the airways, where they can aggravate allergic asthma symptoms. However, the cellular mechanisms by which inhalable particulate allergens (pAgs) potentiate asthmatic symptoms remain unknown, in part because most in vivo and in vitro studies exploring the pathogenesis of allergic asthma use soluble allergens (sAgs). Using a mouse model of allergic asthma, we found that, compared with their sAg counterparts, pAgs triggered markedly heightened airway hyperresponsiveness and pulmonary eosinophilia in allergen-sensitized mice. Mast cells (MCs) were implicated in this divergent response, as the differences in airway inflammatory responses provoked by the physical nature of the allergens were attenuated in MC-deficient mice. The pAgs were found to mediate MC-dependent responses by enhancing retention of pAg/IgE/FcεRI complexes within lipid raft–enriched, CD63(+) endocytic compartments, which prolonged IgE/FcεRI-initiated signaling and resulted in heightened cytokine responses. These results reveal how the physical attributes of allergens can co-opt MC endocytic circuitry and signaling responses to aggravate pathological responses of allergic asthma in mice. / Dissertation

The cytokine tumor necrosis factor α (TNFα) stimulates the NF-кB, SAPK/JNK, and p38 mitogen-activated protein (MAP) kinase pathways by recruiting Rip1 and Traf2 proteins to the tumor necrosis factor receptor 1 (TNFR1). Genetic studies have revealed that Rip1 links the TNFR1 to the IкB kinase (IKK) complex, whereas Traf2 couples the TNFR1 to the SAPK/JNK cascade. We found TNFα-induced p38 MAP kinase activation and interleukin-6 (IL-6) production is impaired in rip1-/- murine embryonic fibroblasts (MEF) but unaffected in traj2-/- MEF, demonstrating that Rip1 is also a specific mediator of the p38 MAP kinase response to TNFα. Moreover, we demonstrate that endogenous Rip1 associates with the MAP3K, Mekk3 in response to TNFα and that TNFα-induced p38 MAP kinase activation is impaired in mekk3-/- cells, indicating that Rip1 may mediate the p38 MAP kinase response to TNFα by recruiting Mekk3. We also demonstrate that Rip1 is phosphorylated and ubiquitinated in response to Tnfα and that Rip1 phosphorylation is not required for ubiquitination of Rip1. Furthermore, TNFα-induced ubiquitination of Rip1 is impaired in Traf2-/- cells, suggesting that Traf2 is the E3 ubiquitin ligase responsible for the TNFα-dependent ubiquitination of Rip1. Finally, recruitment of the ubiquitinated Tak1 complex is dependent on the presence of Rip1, suggesting that Rip1 ubiquitination rather than its phosphorylation is critical in TNFR1 signaling.

Recent studies have revealed extensive cortical demyelination in patients with progressive multiple sclerosis (MS). Demyelination in gray matter lesions is associated with activation of microglia. Macrophages and microglia are known to express myeloperoxidase (MPO) and generate reactive oxygen species during myelin phagocytosis in the white matter. In the present study we examined the extent of microglial activation in the cerebral cortex and the relationship of microglial activation and MPO activity to cortical demyelination. Twenty-one cases of neuropathologically confirmed multiple sclerosis, with 34 cortical lesions, were used to assess microglial activation. HLA-DR immunolabeling of activated microglia was significantly higher in demyelinated MS cortex than control cortex and, within the MS cohort, was significantly greater within cortical lesions than in matched non-demyelinated areas of cortex. In homogenates of MS cortex, cortical demyelination was associated with significantly elevated MPO activity. Immunohistochemistry revealed MPO in CD68-positive microglia within cortical plaques, particularly toward the edge of the plaques, but not in microglia in adjacent non-demyelinated cortex. Cortical demyelination in MS is associated with increased activity of MPO, which is expressed by a CD68-positive subset of activated microglia, suggesting that microglial production of reactive oxygen species is likely to be involved in cortical demyelination.