Made available in DSpace on 2014-12-17T15:32:31Z (GMT). No. of bitstreams: 1 AnaRLAM_TESE.pdf: 3774507 bytes, checksum: 2b9028dcadafe1a4c66e01ba9daab791 (MD5) Previous issue date: 2012-07-19 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Squamous cell carcinoma of the lower lip is among the most common malignant tumors of the oral and maxillofacial region, with good prognosis in more than 90% of patients with 5-year survival. In these carcinomas, the development of lymph node metastasis decreases the prognosis and it has been associated with the formation of new lymphatic vessels. It has been suggested the important role of vascular endothelial growth factor-C (VEGF-C), the receptor type 3 VEGF (VEGFR-3) and hypoxia-induced factor 1 (HIF-1) in this process. The aim of this study was to evaluate the immunoexpression of VEGF-C, VEGFR-3 and HIF-1? and correlate with intra and peritumoral lymphatic density in squamous cell carcinomas of the lower lip metastatic and non-metastatic. The sample consisted of 50 cases of squamous cell carcinoma of lower lip, of which 25 had regional lymph node metastasis and 25, absence of metastasis. The percentages of cells immunostained for VEGF-C, VEGFR-3 and HIF-1? in front of tumor invasion and in the center of tumor were evaluated. Microvessel density lymphatic (MDL) was determined by the counting of lymph microvessels immunostained by the anti-D2-40 in five fields (200?), in an area of evaluation with 0.7386 mm2. The invasion of the lymph vessels by malignant cells was also evaluated. Immunostaining was correlated with the presence and absence of metastasis, TNM clinical stage, local recurrence, disease outcome (remission of injury or patient death) and histological grading. The analysis of intra and peritumoral lymphatic density showed no significant association with clinicopathological parameters and immunoexpressions of VEGF-C, VEGFR-3 and HIF-1? (p > 0,05). There was a weak positive correlation, significant, between intra and peritumoral lymphatic density (r = 0,405; p = 0,004). VEGF-C showed no significant association with clinicopathological and prognosis parameters (p > 0,05). For VEGFR-3, there was scarce membrane staining and intense and homogenous cytoplasmic staining in neoplastic cells. Percentage of positive cytoplasmic VEGFR-3 in center of tumor, exhibited a statistically significant association with metastasis (p = 0,009), patient death (p = 0,008) and histological grades of malignancy proposed by Bryne et al. (1992) (p = 0,002) and World Health Organization (p = 0,003). A low positive correlation was statistically significant between the immunoreactivity of VEGFC and VEGFR-3 cytoplasmic (r = 0,358; p = 0,011) and between the percentage of positive cytoplasmic VEGFR-3 in front of tumor invasion and in the center of the tumor (r = 0,387; p = 0,005) was also demonstrated. There was no association between HIF-1?, clinicopathological and prognosis parameters, and VEGF-C and VEGFR-3. The percentage of nuclear positivity for HIF-1? was significantly higher in cases without invasion of peritumoral lymphatic (p = 0,040). Based on the results we can conclude that most cytoplasmic expression of VEGFR-3 in center of tumor in metastatic cases, high degree of malignancy and poorly differentiated, contributes to poor outcome of squamous cell carcinoma of the lower lip, including patient death. Intra and peritumoral lymphatic density seems to be not associated with lymph node metastasis in these carcinomas / O carcinoma epiderm?ide de l?bio inferior est? entre as les?es malignas mais comuns da regi?o oral e maxilofacial, com progn?stico bom, em mais de 90% dos pacientes com sobrevida de 5 anos. Nestas les?es, o desenvolvimento de met?stase linfonodal diminui sobremaneira o progn?stico e tem sido associado ? forma??o de novos vasos linf?ticos. Tem sido sugerido o importante papel do fator de crescimento endotelial vascular-C (VEGF-C), do receptor tipo 3 do VEGF (VEGFR-3) e do fator 1 induzido por hip?xia (HIF-1) neste processo. O objetivo desta pesquisa foi avaliar as imunoexpress?es de VEGF-C, VEGFR-3, HIF-1? e a densidade linf?tica intra e peritumoral em carcinomas epiderm?ides de l?bio inferior metast?ticos e n?o-metast?ticos, correlacionando-as com par?metros clinicopatol?gicos e progn?sticos. A amostra foi constitu?da por 50 casos de carcinoma epiderm?ide de l?bio inferior, 25 com met?stase linfonodal regional e 25 sem met?stase. Foram avaliados os percentuais de c?lulas imunomarcadas para os anticorpos anti-VEGF-C, anti-VEGFR-3 e anti-HIF-1?, no front de invas?o e no centro tumoral. A densidade microvascular linf?tica (LMVD) foi estabelecida por meio da soma da contagem de microvasos linf?ticos imunomarcados pelo anticorpo anti-D2-40, em cinco campos (200?), em uma ?rea de avalia??o com 0,7386 mm2. A invas?o dos vasos linf?ticos por c?lulas neopl?sicas tamb?m foi avaliada. A imunomarca??o foi relacionada com a presen?a e aus?ncia de met?stase, estadiamento cl?nico TNM, recidiva local, desfecho da doen?a (remiss?o da les?o ou ?bito dos pacientes) e grada??o histol?gica. A an?lise das densidades linf?ticas intra e peritumorais n?o demonstrou associa??o significativa com os par?metros clinicopatol?gicos, progn?sticos e imunoexpress?es de VEGF-C, VEGFR-3 e HIF-1? (p > 0,05). Houve fraca correla??o positiva, significativa, entre as densidades linf?ticas intra e peritumorais (r = 0,405; p = 0,004). O VEGF-C n?o exibiu associa??o significativa entre os par?metros clinicopatol?gicos e progn?sticos avaliados (p > 0,05). Para o VEGFR-3, houve escassa marca??o membranar e intensa e homog?nea marca??o citoplasm?tica nas c?lulas neopl?sicas. O percentual de positividade citoplasm?tica do VEGFR-3, no centro tumoral, exibiu associa??o estatisticamente significativa com a presen?a de met?stase (p = 0,009), ?bito dos pacientes (p = 0,008) e grada??es histol?gicas de malignidade proposta por Bryne et al. (1992) (p = 0,002) e pela Organiza??o Mundial de Sa?de (p = 0,003). Uma fraca correla??o, estatisticamente significativa, entre a imunoexpress?o de VEGF-C e VEGFR-3 citoplasm?tica (r = 0,358; p = 0,011) e entre os percentuais de positividade citoplasm?tica de VEGFR-3 no front de invas?o e no centro tumoral (r = 0,387; p = 0,005) tamb?m foi demonstrada. N?o foi observada associa??o entre o HIF-1? os par?metros clinicopatol?gicos, progn?sticos e o VEGF-C e VEGFR-3. O percentual de positividade nuclear para HIF-1? foi significativamente maior nos casos sem invas?o dos linf?ticos peritumorais (p = 0,040). Com base nos resultados pode-se concluir que a maior express?o citoplasm?tica de VEGFR-3, no centro tumoral, nos casos metast?ticos, de alto grau de malignidade e pobremente diferenciados, contribui para pior evolu??o dos carcinomas epiderm?ides de l?bio inferior, incluindo o ?bito dos pacientes. As densidades linf?ticas intra e peritumorais parecem n?o estar associadas ao densenvolvimento de met?stase linfonodal nestes carcinomas

Einleitung: Ziel dieser Arbeit war es, das RENCA-Nierenzell-Karzinommodell anhand der Hypoxie-induzierten Nekrosen, der Makrophageninfitration sowie der iNOS-Expression und –aktivität zu charakterisieren. Methoden: Für die Erzeugung lokaler Tumoren, wurden Balb/c-Mäusen in vitro kultivierte RENCA-Zellen subkutan appliziert. Die Quantifizierung des Makrophageninfiltrates sowie die Charakterisierung der Hypoxie im Tumorgewebe erfolgten mittels immunhistochemischer Färbungen. Für die Darstellung der Stickstoffmonoxid-Aktivität kamen neben der Immunhistochemie die Reverse Transkriptase Polymerase-Kettenreaktion (RT-PCR) zur Anwendung. Der Nitrit-Gehalt der Tumorproben wurde mittels Griess-Reaktion bestimmt. Ergebnisse: Spontan hypoxische Tumornekrosen sind assoziiert mit einer signifikant geringeren Gefäßdichte. Das Ausmaß der Nekrosen korreliert mit dem Alter der Tumoren. Alte Tumoren (28 Tage) zeigen eine höhere Makrophageninfiltration als junge Tumoren (12 bis 19 Tage). Obwohl die iNOS-Expression auf Proteinebene keinen signifikanten Unterschied zwischen alten und jungen Tumoren ergab, findet sich auf mRNA-Ebene eine signifikant erhöhte iNOS-Expression in alten Tumoren. Hinsichtlich der αActinin-4-Expression konnte kein signifikanter Unterschied gesehen werden. Schlussfolgerung: Bei annähernd gleicher iNOS- und αActinin-4-Expression beziehungsweise erhöhter iNOS-mRNA-Expression ist die Konzentration an Nitrit in Tumoren mit hypoxischer Nekrose reduziert. Daraus kann man schließen, dass die Aktivität der NO-Synthase unter hypoxischen Bedingungen reduziert ist. Dies stützt wiederum die Vermutung, einer die iNOS-Aktivität steuernden normoxie-abhängigen Assoziation von αActinin-4 und iNOS. Die Aktivitätsinhibierung der iNOS könnte somit ein Mechanismus sein, durch den Hypoxie die Zytotoxizität von TAM inhibiert. Die abnehmende Gefäßdichte mit zunehmendem Tumoralter könnte möglicherweise auf eine Regression der Tumorgefäße zurückzuführen sein. Denkbar wäre, dass die Gefäßregression durch makrophagenabhängige Zytokine begünstigt wird.

Indiana University-Purdue University Indianapolis (IUPUI) / Photoacoustic Imaging is a non-invasive optical imaging modality used to image biological tissues. In this method, a pulsating laser illuminates a region of tissues to be imaged, which then generates an acoustic wave due to thermal volume expansion. This wave is then sensed using an acoustic sensor such as a piezoelectric transducer and the resultant signal is converted into an imaging using the back projection algorithm. Since different types of tissues have different photo-acoustic properties, this imaging modality can be used for imaging different types of tissues and bodily organ systems. This study aims at quantifying the process of light conversion into the acoustic signal. Light travels through tissues and gets attenuated (scattered or absorbed) or reflected depending on the optical properties of the tissues. The process of light propagation through tissues is studied using Monte Carlo simulation software which predicts the propagation of light through tissues of various shapes and with different optical properties. This simulation gives the resultant energy distribution due to light absorption and scattering on a voxel by voxel basis. The Monte Carlo code alone is not sufficient to validate the photon propagation. The success of the Monte Carlo code depends on accurate prediction of the optical properties of the tissues. It also depends on accurately depicting tissue boundaries and thus the resolution of the imaging space. Hence, a validation algorithm has been designed so as to recover the optical properties of the tissues which are imaged and to successfully validate the simulation results. The accuracy of the validation code is studied for various optical properties and boundary conditions. The results are then compared and validated with real time images obtained from the photoacoustic scanner. The various parameters for the successful validation of Monte Carlo method are studied and presented. This study is then validated using the algorithm to study the conversion of light to sound. Thus it is a significant step in the quantification of the photoacoustic effect so as to accurately predict tissue properties.