O presente estudo teve como objetivo caracterizar a expressão da família SIBLING (Small Integrin-Binding Ligand, N-linked Glycoproteins) após tratamento regenerativo de furca com enxerto de tecido reparativo de alvéolos dentários. Para isto, os 2os e 3os pré-molares superiores foram extraídos em quatro cães s.r.d. Cinco dias após as extrações, defeitos padronizados de furca classe II foram criados nos 2os, 3os e 4os pré-molares inferiores, bilateralmente. Estes defeitos foram tratados imediatamente com raspagem, alisamento e polimento corono-radicular (RAPCR) e retalho deslocado coronariamente (RDC) (Grupo Controle) ou com RAPCR + RDC + enxerto de tecido reparativo de alvéolos dentários (Grupo Teste) em um experimento de boca-dividida. Após um período de 6 semanas de reparação, os animais foram sacrificados e foi realizada análise imuno-histoquímica para avaliar a localização dos membros da família de proteínas SIBLING, composta pelas seguintes proteínas nãocolágenas da matriz extracelular: osteopontina (OPN), sialoproteína óssea (BSP), proteína da matriz dentinária 1 (DMP1), sialofosfoproteína da dentina (DSPP) e fosfoglicoproteína da matriz extracelular (MEPE). Não foram encontradas diferenças na expressão da família SIBLING entre os grupos teste e controle. Todas as proteínas foram expressas no novo osso, novo cemento e novo ligamento periodontal, em ambos os grupos. Os osteoclastos demonstraram imunolocalização intracelular intensa somente para a OPN. Cementócitos e o novo ligamento periodontal demonstraram, particularmente, marcação intensa para a MEPE. Houve uma diferença evidente entre o padrão de marcação entre o lado tratado (vestibular) e o não-tratado (lingual) de todos os espécimes, com presença de maior marcação do lado vestibular, para todos os anticorpos testados. Podemos concluir que não houve diferenças no padrão de expressão da família SIBLING após o uso do enxerto de tecido reparativo de alvéolos dentários. A família de proteínas SIBLING é expressa durante o processo de reparação de defeitos de furca, indicando possíveis papéis e funções para as proteínas OPN, BSP, DMP1, DSP e MEPE como moléculas alvo em terapias de regeneração periodontal. / The present study aimed in characterizing the expression of the SIBLING (Small Integrin- Binding Ligand, N-linked Glycoproteins) family in a regenerative treatment of furcation defects with a reparative tissue graft obtained from extraction sockets. The second and third upper premolars were extracted in four mixed breed dogs. Five days later, standardized class II furcation defects were created in the second, third and fourth mandibular premolars, bilaterally. The defects were immediately treated with either debridement and root planning (DRP) combined with a coronally positioned flap (CPF) (Control Group), or with DRP+CPF + a reparative tissue graft derived from the second and third premolar extraction sockets (Experimental Group) in a split-mouth design. After 6 weeks period of healing, the animals were sacrificed and immunohistochemistry was carried out to assess the localization of members of the SIBLING family of noncollagenous extracellular matrix proteins, namely osteopontin (OPN), bone sialoprotein (BSP), dentin matrix protein 1 (DMP1), dentin sialophosphoprotein (DSPP) and matrix extracellular phosphoglycoprotein (MEPE). No differences in the SIBLING family of proteins expression were noted between the control and experimental group. All proteins were expressed in new bone, new cementum and new periodontal ligament in both groups. Osteoclasts exhibited intense intracellular localization only for OPN. Cementocytes and the newly formed periodontal ligament demonstrated particularly intense staining for MEPE. There was an evident difference between the staining pattern between the treated (buccal side) and non-treated (lingual) side of the specimens, with a more intense staining pattern in the buccal side, for all the tested antibodies. In conclusion, there were no differences in the pattern of SIBLING expression following the use of a reparative tissue graft obtained from extraction sockets. The SIBLING family of proteins is expressed during the healing process of furcation defects indicating possible roles and functions of OPN, BSP, DMP1, DSPP and MEPE as target molecules in periodontal regeneration therapies.

O projeto tem como objetivo a detecção e caracterização das células epiteliais circulantes de pacientes com carcinoma de mama no estádio III. Analisou-se a presença ou ausência destas células antes da realização de quimioterapia neoadjuvante. Para tanto, utilizou-se o sistema de enriquecimento de amostras de sangue periférico por meio da marcação intracelular imunomagnética direta, empregando-se microbeads conjugados com citoqueratina e posterior separação imunomagnética. A amostra final, com as células alvo detectadas, foi então quantificada por imunocitoquímica ou citometria de fluxo. Resultados: A reação de imunocitoquímica no sangue periférico foi negativa em todos os casos enquanto que na citometria de fluxo foi positiva em 22/23 casos analisados. Conclusão: A detecção e caracterização de células tumorais circulantes em pacientes com câncer de mama podem vir a se constituir em um novo fator prognóstico, porém, são ainda necessárias novas estratégias para melhor detectá-las. / The aim of this study was to detect and characterize carcinoma cells of patients with breast cancer with clinical stage III analyzing the presence or absence before chemotherapy by enrichment of peripheral blood with super paramagnetic microbeads, using direct Immunomagnetic labeling of intracellular 7/8 cytokeratin. For enrichment, magnetically labeled tumor cells are passed over a high-gradient magnetic positive selection column. Cytospin preparations are made after immunomagnetic enrichment and are examined immunocytochemically (ICC) using an anti-cytokeratin and quantified by flow cytometry (CMF). Results: No cytokeratin positive cells were detected in the 32 peripheral blood of patients of breast carcinoma by ICC and cytokeratin positive cells were detected in the 22 of 23 patients by CMF. Conclusion: These methods of detection and characterization breast carcinoma cells will become useful in the diagnosis and prognosis but is necessary optimization of existing methods.

Orientador: José Luiz Laus / Banca: Aline Adriana Bolzan / Banca: Fabrício Villela Mamede / Banca: Delphim da Graça Macoris / Banca: Marcia Rita Fernandes Machado / Resumo: A doença endógena inflamatória uveal em equinos constitui resposta imunopatogênica complexa, onde se admite a participação de uma variedade de antígenos. A uveíte recorrente dos equinos (URE), também denominada ophthalmia periódica ou moon blindness, é uma panuveíte e constitui-se na causa mais comum de diminuição da percepção visual, cuja patogênese permanece, ainda, sob investigação. Inúmeras pesquisas apontam para a hipótese de hipersensibilidade a antígenos bacterianos. A maioria dos estudos mostra evidências de infecção por Leptospira sp.. Examinaram-se, ao acaso, os olhos de 29 animais e coletaram-se o soro, o humor aquoso e o corpo vítreo para proteinograma e aglutinação microscópica para Leptospira sp.. Outrossim, fragmentos de córnea, íris, retina e coróide para histopatologia e imunoistoquímica. A prova de aglutinação microscópica identificou 14 animais positivos, seis animais com titulação igual a 40 e nove indivíduos negativos para as amostras de soro. Houve um animal positivo na amostra de humor aquoso e na de corpo vítreo e outro negativo na de soro (titulação 40), mas positivo na de corpo vítreo. Foram encontrados os sorovares icterohaemorrhagiae, autumalis, patoc, sentot, habdomadis. Foram identificadas, à eletroforese do soro, as proteínas: imunoglobulina A; ceruloplasmina; trasnferrina; hemopexina; albumina; anti-tripsina; imunoglobulina G de cadeia pesada; haptoglobina; glicoproteína ácida; imunoglobulina G de cadeia leve e proteína de 25kda. A única proteína que mostrou resultado estatístico significativo foi a ceruplasmina (p=0,05) com animais soropositivos para leptospirose. À histopatologia a espessura da córnea foi significativamente maior nos animais soropositivos (p=0,0347). O exame imunistoquímico para pesquisa da bactéria Leptospira sp. mostrou maior... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Endogenous uveal inflammatory disease in the horse represent a complex immunopathological response to a range of presumed antigens. Equine recurrent uveitis (ERU) is a panuveitis and the most commmon cause of blindness in horses. Many researchers favour the hypothesis that ERU is delayed type hypersensitivity response to bacterial antigens. New findings show evidence of leptospiral infection in ERU eyes. Fifity eight eyeballs from twenty nine horses randomly selected were studied using histopathology and immunohistochemistry. The serum from 29 animals, aqueous humor an vitreous body from 58 eyes were collected for proteinogram and microscopic agglutination test (MAT) for Leptospira sp.. The microscopic agluttination test was identified 14 positive animals, six with titer 40 and nine negative in serum samples. One seropositive animal was positive in aquaeous humor and vitreous body and another seronegative horse was positive in vitreous body. Were found five serovars from 26 studied from Leptospira interrogans: icterohaemorrhagiae, autumalis, patoc, sentot, habdomadis. Were identified by serum electrophoresis, the following proteins: immunoglobulin A; ceruloplasmin; trasnferrina; hemopexina; albumin, anti-trypsin, the heavy chain of immunoglobulin G; haptoglobin; glycoprotein, immunoglobulin G and the light chain protein, 25kda. The only protein that showed significant results with seropositive horses was ceruplasmin (p = 0.05). The pathomorphologic changes on HE-stained sections showed that seropositive animals for leptospirosis had corneal thickness significantly higher than the seronegative (p = 0.0347). The immunohistochemistry test for Leptospira sp. showed higher positivity in samples of cornea, and some animals were seronegative but positive for this test. At immunohistochemistry was observed that the anti-metalloproteinase... (Complete abstract click electronic access below) / Doutor

As doenças periodontais (DPs) são alterações inflamatórias crônicas que acometem os tecidos de sustentação do órgão dental. A presença do diabetes é refletida em maior severidade e prevalência das DPs tanto em humanos quanto em modelos experimentais. Contudo, os mecanismos biológicos envolvidos no aumento da prevalência e da severidade permanecem pouco conhecidos. Desta forma, o objetivo deste estudo foi avaliar o número de células marcadas por imunohistoquímica para TNF-&#x3B1;, RANKL, OPG, IL-10 e para o fator de transcrição RUNX-2, na doença periodontal experimental decorrente da indução do diabetes em ratos. Além disso, avaliamos os possíveis efeitos do tratamento com chá verde sobre o periodonto dos animais. Inicialmente, os ratos (n=80) foram submetidos à indução do diabetes por administração intraperitoneal de estreptozotocina (50mg/kg) e, juntamente com o grupo controle (n=40), foram subdivididos em animais tratados com chá verde ou com água, acompanhados durante o período de 15, 30, 60 ou 90 dias. Após o sacrifício dentro do tempo determinado para cada grupo, as hemimaxilas coletadas passaram pelos procedimentos de imunohistoquímica. Os resultados revelaram que a presença do diabetes causou perda óssea alveolar, compatível com o desenvolvimento da doença periodontal e resultou em alterações significativas no número de células imunomarcadas para diferentes mediadores do processo inflamatório. Entretanto, o chá verde apresentou efeitos benéficos para o periodonto, alterando a marcação das citocinas envolvidas. Nos animais diabéticos, independente do tratamento, foi observado aumento estatisticamente significativo (p<0,05 ANOVA) no número de células imunomarcadas para TNF-&#x3B1; e RANKL. Inversamente, houve menor marcação para OPG (60 e 90 dias), RUNX-2 (30, 60 e 90 dias) e para IL-10 (30, 60 e 90 dias) nos animais que ingeriram água. Porém, os diabéticos tratados com chá não demonstraram diferenças significativas em relação ao seu respectivo controle. Assim, quando comparados os grupos diabéticos tratados com água e chá, os animais que receberam o chá demonstraram significativamente (p<0,05 ANOVA) menor quantidade de células munomarcadas para TNF-&#x3B1; e RANKL em relação aqueles diabéticos tratados com água, enquanto que a marcação para OPG, RUNX-2 e IL-10 foi maior. Portanto, a desregulação na expressão de citocinas e do fator de transcrição osteoblástico parece ser um dos mecanismos biológicos envolvidos no aumento da prevalência e da severidade das doenças periodontais decorrentes do diabetes. Entretanto, o chá verde modulou a inflamação no periodonto dos animais diabéticos, apresentando-se, possivelmente, como uma alternativa terapêutica e coadjuvante ao tratamento das doenças periodontais correlacionadas ao diabetes. / Periodontal diseases (PD) are chronic inflammatory diseases leading the destruction of connective tissue and alveolar bone supporting the teeth. The establishment of diabetes increases PD prevalence and severity in humans and experimental model. However, biological mechanisms regarding to increase of prevalence and severity remains poorly known. The aim of this study was to evaluate the number of immuno-staining cells to TNF-&#x3B1;, RANKL, OPG, IL-10 and transcription factor RUNX-2 in experimental periodontal disease in diabetic rats. Furthermore, the possible green tea efects were evaluated in periodontiumof the rats. Diabetes was induced in Wistar rats (n=120) by intraperitoneal administration of 50 mg/kg ofstreptozotocin and together with control animals (n=80), the rats were subdivided in water or green tea treated group, that were analyzed at 15, 30, 60 and 90 days after diabetes induction. The animals were sacrificed and the hemimaxillae were removed and submitted to immunohistochemistry procedures. Our data demonstrated that diabetes induction and progression resulted in significant bone loss and alterations in number of immuno-staining cells to different mediators of inflammatory process. However, the green tea showed positive effects in periodontium through inflammation modulation. In diabetic rats, regardless of treatment, we observed an increased number of immuno-staining cells to TNF-&#x3B1;, IL-1b and RANKL (p<0,05 ANOVA). On the other hand, in water treated diabetic rats, there were a decreased number of immuno-staining cells to OPG (60 e 90 days), RUNX-2 (30, 60 e 90 days) and IL-10 (30, 60 e 90 days). However, the green tea treated rats did not showed statistical differences between control and experimental groups in those staining. When we compared both diabetic groups, green tea and water treated, the animals that drank the green tea showed decreased number of immuno-staining cells to TNF-&#x3B1; and RANKL(p<0,05 ANOVA) whereas the number of immuno-staining cells to OPG, RUNX-2 e IL-10 were increased. Taken together, desregulation of inflammatory cytokines and osteoblastic transcripition factor seem to be one of biological mechanisms involved in the increase of periodontal disease prevalence and severity associated with diabetes. However the green tea was able to modulate inflammation in diabetic rats periodontium and it seems to be a possible terapeutic agent to periodontal disease treatment associated with diabetes.

O estabelecimento e perfeito funcionamento da placenta são fatores dependentes da intensa vascularização ocorrida no órgão. Os processos de vasculogênese e angiogênese placentária são modulados por diversos fatores, incluindo o VEGF (fator de crescimento vascular endotelial) e bFGF (fator de crescimento fibroblástico básico). Apesar da importância do VEGF e bFGF durante a vascularização estar estabelecida, vários estudos indicam a participação desses fatores de crescimento como moduladores locais em outras funções fisiológicas, como por exemplo o controle da produção hormonal em tecidos esteroidogênicos. Animais clonados podem apresentar alterações na expressão de determinados genes durante seu desenvolvimento, o que pode alterar a função placentária. Os objetivos deste estudo são determinar a localização tecidual do VEGF, bFGF e seus receptores na placenta bovina e avaliar a influência destes fatores de crescimento sobre a produção de progesterona placentária em bovinos não clonados e clonados. Placentomas de 90, 150 e 210 dias de gestação foram obtidos em abatedouro e placentônios de gestações aos 270 dias provenientes de bovinos clonados e não clonados foram coletados após cesarianas. As amostras foram fixadas em formol tamponado 4%, desidratadas e incluídas em parafina. Cortes foram submetidos a imuno-histoquímica para posterior localização das proteínas do VEGF, bFGF e seus receptores. Sob condições assépticas, as células foram mecanicamente dispersas e cultivadas em placas de 96 cavidades. Os fatores foram adicionados em concentrações de 10 e 50 &#951;g/ml de bFGF e VEGF, respectivamente. Amostras de meio de cultura e as células dos grupos controle, bFGF, VEGF e VEGF mais bFGF foram coletadas 24, 48 e 96 horas após a adição dos fatores. A progesterona foi dosada por radioimunoensaio e o conteúdo protéico pelo método de Lowry. Os dados foram analisados utilizando-se o programa estatístico SAS (Statistical Analysis System), as diferenças estatísticas encontradas foram comparadas pelo teste de variação múltipla de Duncan. O VEGF, bFGF e seus receptores foram localizados em células do epitélio e estroma maternos e fetais e células endoteliais vasculares em bovinos não clonados e clonados. As células placentárias apresentaram diferentes capacidades de síntese de progesterona ao longo da gestação. Aos 90 e 210 dias de gestação o VEGF estimulou a produção de progesterona, enquanto aos 270 dias de gestação o fator inibiu a produção deste hormônio. O bFGF estimulou a produção de progesterona pelas células placentárias aos 90 dias de gestação. A adição dos dois fatores de crescimento conjuntamente determinou um estímulo na produção de progesterona aos 210 dias de gestação. A produção de progesterona pelas células de bovinos clonados foi semelhante àquela observada em células de bovinos não clonados na mesma idade gestacional e os fatores de crescimento não influenciaram essa produção. Conclui-se que o VEGF e bFGF, atuando localmente no tecido placentário, funcionam como moduladores do processo de esteroidogênese, influenciando de maneira tempo-dependente a produção de progesterona deste órgão. / Placental establishment and function are dependent on intense vascularization. Placental vasculogenesis and angiogenesis are modulated by several factors, including VEGF (vascular endothelial growth factor) and bFGF (basic fibroblast growth factor). Although the role of VEGF and bFGF during vascularization is already well established, some studies have indicated the participation of these growth factors as local modulators in other physiological functions, such as control of hormonal production in steroidogenic tissues. Cloned animals may exhibit alterations in gene expression during development modifying placental function. The aims of this study are to determine the tissue localization of VEGF, bFGF and their receptors in the bovine placenta and to evaluate the influence of bFGF and VEGF on placental progesterone production in non-cloned and cloned bovines. Placentomes from days 90, 150 and 210 of pregnancy were obtained at local slaughterhouse and placentomes from cloned and non-cloned gestations at 270 days were obtained after cesarean sections. Samples were fixed in 4% buffered formol solution, dehydrated and included in paraffin. Sections were subimitted to immunohistochemistry for subsequent localization of VEGF, bFGF and their receptors proteins. Under aseptic conditions, cells were mechanically dispersed and then cultivated in a 96-well plate. Growth factors were added at concentrations of 10 and 50 &#951;g/ml for bFGF and VEGF, respectively. Samples of culture medium and cells from control, bFGF, VEGF and bFGF plus VEGF groups were collected 24, 48 and 96 hours after growth factor addition. Progesterone concentrations were assessed by radioimmunoassay and protein content was measured by Lowry?s method. Data were analyzed by SAS (Statistical Analysis System) program, significant differences were compared by Duncan?s range multiple test. VEGF, bFGF and their receptors were localized in maternal and fetal epithelial and stromal cells and vascular endothelial cells during pregnancy in non-cloned animals and in cloned bovine placenta at 270 days of pregnancy. Bovine placental cells were able to produce different amounts of progesterone during pregnancy. Growth factors were able to influence progesterone production in placental cells only after 24 hours in culture. At 90 and 210 days of pregnancy VEGF stimulated progesterone production, while at 270 days of pregnancy the growth factor inhibited production of this hormone. bFGF stimulated progesterone production in placental cells from 90 days of pregnancy. Both growth factors together determined an increase in progesterone production in placental cells from 210 days of pregnancy. Progesterone production in placental cells from cloned cattle is similar when compared with non-cloned placental cells at the same gestational age and growth factors did not influence progesterone production in these cells. VEGF and bFGF, acting locally in the placental tissue, are modulators of the steroidogenic process, influencing in a time-dependent manner the progesterone production in this organ.

A matriz extracelular (MEC) pode ser definida como um complexo de proteínas e glicoproteínas que envolve as células nos mais diversos tecidos e tem papel importante na diferenciação e atividade celular, bem como no processo de mineralização e nos processos neoplásicos. Os componentes não colágenos da MEC têm sido abundantemente estudados, visando conhecer os minuciosos detalhes da biologia dos tecidos e assim entender os mecanismos envolvidos em suas patologias. Neste contexto, o presente trabalho tem como objetivo estudar a expressão e distribuição dos seguintes componentes não colágenos da MEC dos tecidos dentais: biglican, decorin, fibromodulin, osteonectina (ONC), osteopontina (OPN), sialoproteína óssea (BSP) e osteocalcina (OCC) no ameloblastoma e no tumor odontogênico cístico calcificante (cisto de Gorlin). Para ta nto foi utilizada a técnica da imunoistoquímica, com o método da estreptavidina-biotina-peroxidase, e anticorpos contra as proteínas anteriormente citadas. Os resultados mostraram que o biglican, o decorin e a BSP foram expressos somente nas células epiteliais metaplásicas, nas células fantasmas e células fantasmas em processo de calcificação, no estroma dos ameloblastomas e no ectomesênquima neoplásico do tumor odontogênico cístico calcificante. Já o fibromodulin e a OC foram predominantemente negativos no componente epitelial e no mesenquimal, com exceção para as células fantasmas, células fantasmas em processo de calcificação e áreas de hialinização próximas ao epitélio. A ONC foi positiva na maioria das células epiteliais, com exceção das células estrelárias dos ameloblastomas folicular e acantomatoso, e também no componente mesenquimal de ambas neoplasias. Já a OPN apresentou positividade somente nos focos de calcificação presentes no tumor odontogênico cístico calcificante. As proteínas estudadas apresentaram distribuição semelhante em neoplasias caracterizadas por padrões de crescimento diferentes, levando a crer que, apesar de participarem ativamente do mecanismo de crescimento neoplásico intra-ósseo, isoladamente não exercem papel decisivo na determinação do tipo de padrão de crescimento. Outro fato digno de relevância é a baixa expressão dessas proteínas nas células epiteliais neoplásicas quando comparada com a expressão no estroma e ectomesênquima, levando-nos a crer que as células epiteliais atuem principalmente como estimuladores da expressão dessas proteínas, que, por sua vez, podem atuar de forma agonista ou antagonista ao crescimento neoplásico. / The extracellular matrix (ECM) can be defined as a complex of proteins and glycoproteins that involves the cells in all tissues. It has a key role in cell differentiation and activity, as well as in mineralization and neoplastic processes. The non-collagenous components of the ECM have been abundantly studied to know the details of the biology of tissues and thus to understand the mechanisms involved in its pathologies. The aim of the present work is to study the expression and distribution of the following noncollagenous components of the ECM of dental tissues: biglycan, decorin, fibromodulin, osteonectin (ONC), osteopontin (OPN), bone sialoprotein (BSP) and osteocalcin (OCC) in ameloblastoma and the calcifying cystic odontogenic tumour. The streptavidin-biotinperoxidase method of immunohistochemistry was used with antibodies against the antigens previously cited. The results show that biglican, decorin and BSP had been expressed only in metaplastic epithelial cells, in ghost cells and ghost cells in calcification process, stroma of ameloblastomas and neoplastic ectomesenchyma of the calcifying cystic odontogenic tumour. The fibromodulin and the OCC showed predominantly negative expression in the epithelial and mesenchymal components, with exception for the ghost cells, ghost cells in calcification process and hyalinization areas next to the epithelium. The ONC was positive in the majority of the epithelial cells, with exception of the central cells of follicular and acanthomatous ameloblastomas, and also in the mesenchymal component of both tumours. OPN presented positivity only in the calcification focus of the calcifying cystic odontogenic tumour. The proteins studied presented similar distribution in tumours characterized by different patterns of growth, leading to believe that although they participate actively of the mechanism of intraosseous growth, separately they do not exert a key role in the determination of the type of growth pattern. Another relevance fact is the low expression of these proteins in the neoplastic epithelial cells when compared to the expression in stroma and ectomesenchyma, which make us believe that the epithelial cells act mainly as stimulators of the expression of these proteins, which in turn can act as agonist or antagonist to the tumour growth.

The integrins play a crucial role in cancer cell proliferation, migration, differentiation, survival and angiogenesis. It has been shown that integrin expression is positively correlated to cancer dissemination, this suggests targeting selected integrins as an anti-metastatic strategy. The aim of this study is to investigate the effect of novel antagonists of α5β1, αvβ3 and αvβ5 integrins on cancer cell migration, a key process in tumour cell dissemination. Immunohistochemistry was used to evaluate the expression of α5, αv, β3 and β5 integrin subunits in prostate cancer tissues. Furthermore the expression of these integrin subunits in tumour and normal human head and neck tissues was compared. The expression profile of these integrin subunits in established human cancer cell lines was subsequently evaluated using immunodetection methods in cells and xenograft tumour samples. The effect of integrin inhibition on cell migration was then assessed using neutralizing antibodies against αvβ3, αvβ5, and α5β1 integrins in the scratch-wound healing assay. This assay was then used to evaluate the potential of novel small molecule integrin antagonists in preventing tumour cell migration. In H & N tissues, αvβ3, αvβ5 and α5β1 integrins are extensively expressed in tumour tissues but weakly expressed in normal tissue from the same patient. Further, prostate cancer tissues expressed variable levels of αvβ3, αvβ5 and α5β1 integrins. αvβ3 and αvβ5 integrins were expressed in variable levels in OSC-19, PC-3, DU145, DLD-1, HT-29, HUVEC, MCF-7, MCF-7ADR and M14 human tumour cell lines and in OSC-19, PC-3, HT-29 and MCF-7 xenografts. α5β1 integrin was expressed in all cell lines and xenografts except in MCF-7 cell line and HT-29 cell line and xenograft. Overall, the expression was elevated in xenografts compared to the corresponding cultured cells. Based on the expression profile and ability of cells to migrate, three cell lines (DLD-1 colon, DU145 prostate and OSC-19 HNSCC) were selected as models to further evaluate the potential of novel small molecule integrin antagonists to inhibit cell migration. The cell lines were characterized by using neutralizing antibodies against αvβ3, αvβ5, and α5β1 integrins to determine which of these three integrins were primarily involved in tumour cell migration. In DLD-1 and DU145, blocking αvβ5 and αvβ3 significantly inhibited migration, whilst the migration of OSC-19 was 50% inhibited by a multi-integrin inhibitor combination. Among the antagonists, ICT9055 and ICT9072 significantly decreased DLD-1 cell migration by 70% and 60% respectively while ICT9023, ICT9024, and ICT9026 significantly decreased DU145 cell migration by 60%, 60% and 50% respectively. The findings suggest that single integrin inhibition is not sufficient to prevent cell migration whereas dual or multiple inhibition is more effective. Two novel anti-migratory agents were identified in colon cancer and three in prostate cancer which would warrant further investigation.

Chen Lei. / "February 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 124-161). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Introdução: Proteínas da Superfamília do fator transformador de crescimento β (TGF-β) estão implicadas na regulação de diversas funções biológicas. Embora alguns estudos revelaram a sua presença no endométrio ectópico de portadoras de adenomiose, a sua função na etiopatogenia da doença permanece pouco conhecida. Objetivo: o estudo visa comparar a expressão imuno-histoquímica de TGF-β1 no endométrio ectópico de portadoras de adenomiose com o endométrio tópico de pacientes sem essa condição. Método: Estudo de caso-controle utilizando imuno-histoquímica em amostras uterinas (blocos de parafina) do Hospital de Clínicas de Porto Alegre. A amostra contém 28 casos de adenomiose e 21 controles. Resultados: Não encontramos associação entre tabagismo e adenomiose (P = 0,75), abortos e adenomiose (P = 0,29), gestações e adenomiose (P = 0,85), curetagens e adenomiose (P = 0,81), dor pélvica e adenomiose (P = 0,72) e presença de mioma e adenomiose (P = 0,15). Além disso encontramos relação entre sangramento uterino anormal (SUA) e adenomiose (P = 0,02) e cesarianas prévias e adenomiose (P = 0,02) . A expressão imuno-histoquímica de TGF-β1 no endométrio ectópico de portadoras de adenomiose não teve diferença significativa quando comparado com a expressão dessa proteína no endométrio tópico de pacientes sem adenomiose (P = 0,86). Conclusão: Nosso estudo foi um dos primeiros a comparar a expressão de TGF-β1 no endométrio de pacientes com e sem adenomiose. Em nossa análise não obtivemos diferença significativa entre os grupos, resultado diferente do encontrado em outros dois estudos. Mais estudos são necessários para investigar o papel da superfamília TGF no desenvolvimento e manutenção da adenomiose. / Background: Proteins of transforming growth factor β superfamily (TGF-β) are implicated in the regulation of various biological functions. Although some studies have revealed their presence in ectopic endometrium of women with adenomyosis, their role in the pathogenesis of the disease remains largely unknown. Objective: The study aims to compare the immunohistochemical expression of TGF- β1 in ectopic endometrium of women with adenomyosis with the topic endometrium of patients without this condition. Methods: Casecontrol study using immunohistochemistry in uterine samples (paraffin blocks) obteined from Hospital de Clínicas de Porto Alegre. The sample contained 28 adenomyosis cases and 21 controls. Results: We found no significant difference between smoking and adenomyosis (P = 0.75), abortions and adenomyosis (P = 0.29), pregnancies and adenomyosis (P = 0.85), curettage and adenomyosis (P = 0.81), pelvic pain and adenomyosis (P = 0.72) and presence of myoma and adenomyosis (P = 0.15). We did find a relationship between adenomyosis and abnormal uterine bleeding (AUB) (P = 0.02) and previous cesarean section and adenomyosis (P = 0.02). Immunohistochemical expression of TGF-β1 in ectopic endometrium of women with adenomyosis did not have significant difference when compared with the expression of this protein in the topic endometrium of patients without adenomyosis (P = 0.86). Conclusion: Our study was one of the first to compare the TGF-β1 expression in the endometrium of patients with and without adenomyosis. In our analysis we have not had significant difference between the groups, unlike observed in two other studies. More studies are needed to investigate the role of TGF superfamily in the development and maintenance of adenomyosis.

by Wang Wen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 113-124). / Abstracts in English and Chinese. / ABSTRACT --- p.i / FLOWCHART --- p.vi / ACKNOWLEDGEMENT --- p.x / ABBREVIATIONS --- p.xi / INDEX FOR FIGURES --- p.xii / INDEX FOR TABLES --- p.xv / TABLE OF CONTENTS --- p.xvi / Chapter 1. --- INTRODUCTION --- p.1 / Chapter 1.1 --- PATELLAR TENDINOSIS --- p.1 / Chapter 1.1.1 --- Introduction --- p.1 / Chapter 1.1.2 --- Epidemiology of Patellar Tendinosis --- p.3 / Chapter 1.1.3 --- Etiology of Patellar Tendinosis --- p.3 / Chapter 1.1.4 --- Manifestations of Patellar Tendinosis --- p.4 / Chapter 1.1.5 --- Imaging Examination on Patellar Tendinosis --- p.4 / Chapter 1.1.6 --- Clinical Diagnosis of Patellar Tendinosis --- p.6 / Chapter 1.1.7 --- Management of Patellar Tendinosis … --- p.6 / Chapter 1.2 --- ANATOMY AND HISTOLOGY OF PATELLAR TCNDON --- p.7 / Chapter 1.3 --- STRUCTURE AND METABOLISM OF TENDON --- p.9 / Chapter 1.3.1 --- Tenocytes --- p.9 / Chapter 1.3.2 --- Extra-cellular Matrix --- p.11 / Chapter 1.3.2.1 --- Collagen --- p.11 / Chapter 1.3.2.2 --- Proteoglycans --- p.12 / Chapter 1.4 --- ROLES OF GROWTH FACTORS TENDON HEALING AND REPAIR --- p.14 / Chapter 1.4.1 --- Platelet-Derived Growth Factor --- p.14 / Chapter 1.4.2 --- Transforming Growth Factor-beta --- p.15 / Chapter 1.5 --- HISTOPATHOLOGY OF PATELLAR TENDINOSIS --- p.16 / Chapter 1.6 --- STUDY PLAN --- p.17 / Chapter 1.6.1 --- Characterization on Hypercellularity --- p.18 / Chapter 1.6.2 --- Characterization on Disorganization and Loosening of Collagen --- p.18 / Chapter 1.6.3 --- Characterization on Inflammatory Trace --- p.20 / Chapter 1.6.4 --- Characterization on Growth Factors in Tendinosis --- p.21 / Chapter 1.7 --- OBJECTIVES --- p.22 / Chapter 2. --- MATERIALS AND METHODS --- p.27 / Chapter 2.1 --- HUMAN TISSUES --- p.27 / Chapter 2.1.1 --- Patellar Tendinosis Tissues --- p.27 / Chapter 2.1.1.1 --- Diagnosis of patellar tendinosis --- p.27 / Chapter 2.1.1.2 --- Recruitment of patients --- p.27 / Chapter 2.1.4 --- Healthy Patellar Tendon tissues --- p.28 / Chapter 2.2 --- TISSUES COLLECTION AND PREPARATION --- p.28 / Chapter 2.3 --- HISTOLOGICAL STUDY ON HUMAN SPECIMENS --- p.28 / Chapter 2.3.1 --- Haematoxyline and Eosin Staining --- p.29 / Chapter 2.3.2 --- Safranin O Staining --- p.29 / Chapter 2.3.2.1 --- Reagents preparation --- p.29 / Chapter 2.3.2.2 --- Experimental procedure --- p.30 / Chapter 2.3.5 --- Polarization Microscopy --- p.30 / Chapter 2.4 --- IMMUNOHISTOCHEMICAL STAINING --- p.30 / Chapter 2.4.1 --- Reagents Preparation --- p.31 / Chapter 2.4.2 --- Experimental Procedure --- p.33 / Chapter 2.5 --- IMAGE ANALYSIS --- p.35 / Chapter 2.5.1 --- Equipment --- p.35 / Chapter 2.5.2 --- Procedures --- p.35 / Chapter 2.6 --- IN SITU ZYMOGRAPHY --- p.37 / Chapter 2.6.1 --- Reagents Preparation --- p.37 / Chapter 2.6.2 --- Experimental Procedure --- p.38 / Chapter 2.7 --- STATISTIC ANALYSIS.… --- p.39 / Chapter 3. --- RESULTS --- p.42 / Chapter 3.1 --- HUMAN SAMPLES --- p.42 / Chapter 3.1.1 --- Patellar tendinosis patients --- p.42 / Chapter 3.1.2 --- Healthy control group --- p.43 / Chapter 3.2 --- HISTOLOGICAL STUDY ON HUMAN SPECIMENS --- p.43 / Chapter 3.2.1 --- Gross Morphology --- p.43 / Chapter 3.2.2 --- Haematoxyline and Eosin Staining --- p.44 / Chapter 3.2.3 --- Safranin O Staining --- p.44 / Chapter 3.2.4 --- Polarization Microscopy --- p.44 / Chapter 3.3 --- IMAGE ANALYSIS --- p.45 / Chapter 3.3.1 --- Immunohistochemistry of PCNA --- p.45 / Chapter 3.3.2 --- Immunohistochemistry of hsp47 --- p.46 / Chapter 3.3.3 --- Immunohistochemistry of Procollogen Type I --- p.47 / Chapter 3.3.4 --- Immunohistochemistry of MMP1 --- p.47 / Chapter 3.3.5 --- Immunohistochemistry of TIMP1 --- p.48 / Chapter 3.3.6 --- Immunohistochemistry of COX-2 --- p.49 / Chapter 3.3.7 --- Immunohistochemistry of TGFP --- p.49 / Chapter 3.3.8 --- Immunohistochemistry of PDGFbb --- p.50 / Chapter 3.3.9 --- Immunohistochemistry of PDGFRβ --- p.51 / Chapter 3.3.10 --- Summary of Image Analysis of Immunohistochemical staining --- p.51 / Chapter 3.4 --- IN SITU ZYMOGRAPHY --- p.52 / Chapter 4. --- DISCUSSION --- p.93 / Chapter 4.1 --- DIAGNOSIS OF PATELLAR TENDINOSIS --- p.93 / Chapter 4.2 --- HYPERCELLULARITY IN PATELLAR TENDINOSIS --- p.95 / Chapter 4.3 --- COLLAGEN DISTURBANCE IN PATELLAR --- p.97 / Chapter 4.4 --- INFLAMMATORY RESPONSE IN PATELLAR TENDINOSIS --- p.100 / Chapter 4.5 --- THE EXPRESSION OF GROWTH FACTORS IN PATELLAR TENDINOSIS --- p.102 / Chapter 4.6 --- PROPOSED PATHOGENESIS FOR PATELLAR TENDINOSIS --- p.105 / Chapter 4.7 --- LIMITATION OF THIS STUDY --- p.108 / Chapter 4.8 --- FUTURE STUDY --- p.109 / Chapter 5. --- CONCLUSION --- p.111 / BIBLIOGRAPHY --- p.113