Made available in DSpace on 2014-07-29T15:07:52Z (GMT). No. of bitstreams: 1 dissertacao leonardo ciencia animal.pdf: 381642 bytes, checksum: 916af2bb18068d3efe78bdc7c8582617 (MD5) Previous issue date: 2009-10-29 / Myxomatous mitral valve degeneration (MMVD) or mitral valve endocardiosis is an endocardial age-related disease characterized by the mucopolysaccharide (MPS) accumulation and by collagen degeneration in mitral valves of many species, especially in humans, dogs and swine. Metalloproteinases (MMP) are proteolytic enzymes responsible for the extracellular matrix (ECM) remodelling in regular and pathological tissue. It has been suggested that the changing of the valvular distribution of these enzymes might be responsible for MMVD genesis. Likewise, vasoactive substances like nitric oxide (NO) played a role in the endocardiosis development. The aim of this study was to evaluate the histological changes, the expression of nitric oxide synthase (NADPH-d) and the immunohistochemical localization of MMP-2 and MMP-9 in the anterior leaflet in dogs and swine with regular mitral valves and those with endocardiosis. For this purpose, two experiments were done. First, 12 mitral valves of dogs and 22 of swine were analyzed. Valves were fixed in a 4% paraformaldehyde, exposed to NADPH-d reaction, processed routinely and microscopically evaluated for the detection of MPS deposition, collagen degeneration and fibrosis. In dogs, very high intensity reaction to NADPH-d was associated with higher endocardiosis degree and with presence of MPS deposition as well as collagen degeneration. There was no alteration in colour during the swine valves reaction to NADPH-d. In conclusion, NO works in canine mitral valves, remodeling MEC and playing a role in dogs mitral endocardiosis disease. In swine, it is suggested that NO has restricted action in MEC or there are major differences on the structures of swine valves because there was no reaction to NADPH-d and absence of macroscopical endocardiosis lesions. For the second study, 25 mitral valves of dogs and 32 valves of swine were also analyzed. Valves were macroscopically evaluated for the occurrence or not of endocardiosis. They were fixed in a 4% paraformaldehyde, routinely processed and submitted to immunohistochemical reaction and microscopically evaluated for the intensity of antigen labelling and for the number of positive cells, as well as MPS deposition, collagen degeneration and fibrosis. In dogs, DMVM is characterized by MPS accumulation. This collagen deposition and degeneration are directly related to the endocardiosis level. The MMP-2 and MMP-9 enzymes are involved in dogs myxomatous mitral valve degeneration process. In swine, even with the lack of microscopical endocardiosis there was observed some MPS deposition changes, especially in females. In these animals valves there are constitutive expression of MMP-2 and MMP-9, what suggests the action of these enzymes in the normal MEC mitral valvular remodeling in young and old female pigs / A degeneração mixomatosa da válvula mitral (DMVM) ou endocardiose mitral é uma doença do endocárdio valvular relacionada com a idade e caracterizada por acúmulo de mucopolissacarídeos (MPS) e degeneração do colágeno na mitral de várias espécies mamíferas, sendo observada com maior frequência em humanos, cães e suínos. Metaloproteinases (MMP) são enzimas proteolíticas dependentes do zinco, responsáveis pelo remodelamento da matriz extracelular (MEC) em processos normais e patológicos de vários tecidos. A alteração na expressão e a distribuição valvular dessas enzimas estão envolvidas na gênese da DMVM. Da mesma forma, substâncias vasoativas como o óxido nítrico (NO) tem papel importante no desenvolvimento da endocardiose. Assim, o objetivo deste estudo foi avaliar as alterações histológicas, a expressão da sintase de óxido nítrico (NADPH-d), e o padrão de marcação imunoistoquímica de MMP-2 e MMP-9 no folheto anterior da válvula mitral normal e com endocardiose de cães, de suínos jovens e matrizes. Para isso, realizaram-se dois experimentos. No primeiro foram utilizadas 12 mitrais de cães adultos a idosos, 22 válvulas de suínos, 10 de animais com idade de sete e oito meses, e 12 mitrais de matrizes entre cinco e seis anos. As válvulas foram avaliadas macroscopicamente quanto ocorrência ou não de endocardiose, colhidas, fixadas em paraformaldeído a 4%, submetidas à reação de NADPH-d, incluídas em parafina e avaliadas microscopicamente quanto à deposição de mucopolissacarídeos (MPS), degeneração do colágeno, fibrose e grau de endocardiose. Nas válvulas caninas quanto maior a intensidade da reação de NADPH-d, maiores eram o grau de endocardiose, a deposição de MPS e a degeneração do colágeno. Nas válvulas suínas não houve coloração à reação de NADPH-d. Com isso, concluiu-se que o NO atua na mitral canina, remodelando a MEC e participando da patogenia da endocardiose mitral nessa espécie. Na mitral suína sugere-se a ação restrita do NO no remodelamento da MEC ou diferenças estruturais na válvula desta espécie, já que não houve reação ao NADPH-d e lesões macroscópicas de endocardiose. No segundo estudo foram utilizadas 25 mitrais de cães adultos e idosos e 32 válvulas de suínos, sendo 10 de animais com idade de sete e oito meses, e 22 mitrais de matrizes entre cinco e seis anos. As válvulas foram avaliadas macroscopicamente quanto ocorrência ou não de endocardiose, fixadas em paraformaldeído a 4%, incluídas em parafina, submetidas à reação de imunoistoquímica e avaliadas microscopicamente quanto à intensidade de marcação e número de células marcadas, quanto à deposição de MPS, degeneração do colágeno e fibrose. Nos cães, concluiu-se que a DMVM é caracterizada por acúmulo de MPS e degeneração do colágeno, que estão diretamente relacionadas ao grau de endocardiose e, consequentemente, à gravidade da doença valvular. Ainda, as enzimas MMP-2 e MMP-9 estão envolvidas na degeneração mixomatosa da válvula canina. Em suínos, mesmo na ausência macroscópica de endocardiose podem ser observadas alterações como deposição de MPS, degeneração do colágeno e fibrose, sendo esta última evidente em matrizes. Também, nesses animais há expressão constitutiva de MMP-2 e MMP-9, sugerindo a participação dessas enzimas no remodelamento da MEC valvular mitral normal em suínos jovens e matrizes

A endometriose é uma doença de origem multifatorial, caracterizada por presença de tecido endometrial fora da cavidade uterina, responsável por sintomas álgicos com grande impacto na qualidade de vida da paciente, além de ser um dos principais fatores de infertilidade. Muitos estudos já foram realizados no intuito de explicar a etiopatogenia da endometriose, assim como muito tem sido estudado para encontrar novas estratégias de tratamento. Várias linhas de medicamentos têm sido estudadas com este intuito, agindo em diferentes pontos da etiopatogênese da doença, uma delas na inibição de metaloproteinases da matriz extracelular, que tem papel no remodelamento do mesotélio do peritônio e angiogênese. O objetivo deste estudo foi avaliar a influência de uma droga (doxiciclina) de baixo custo, com ação conhecida na inibição das metaloproteinases, em endometriose peritoneal induzida em ratas. Para isso, foram usadas 30 ratas adultas Wistar com lesão induzida de endometriose, divididas em três grupos, um grupo controle (C, n=10) sem tratamento, um grupo onde foi administrado doxiciclina em baixa dose (BD, n=10) e um grupo onde foi realizado doxiciclina em alta dose (AD, n=10). Foi realizada avaliação da área das lesões de cada rata e estudo imunohistoquímico para positividade de anticorpo primário de metaloproteinase de matriz 9 (MMP9) e de inibidor de metaloproteinase de matriz 2 (TIMP2). A doxiciclina atuou reduzindo a área das lesões nos grupos BD e AD (p=0,0052) em relação ao grupo C e reduzindo a expressão do TIMP2 no grupo AD (p=0,0009) em relação aos grupos BD e C. Não houve resultado significativo na expressão da MMP9. / Endometriosis is a multifactorial origin disease, characterized by the presence of endometrial tissue outside the uterine cavity, responsible for painful symptoms with important impact on the life quality of the patient, besides being one of the main factors of infertility. Many studies have already been carried out to explain the etiopathogenesis of endometriosis, and much has been studied to find new treatment strategies. Several lines of drugs have been studied for this purpose, acting at different points in the etiopathogenesis of the disease, one of them in the inhibition of extracellular matrix metalloproteinases, which plays a role in the remodeling of the peritoneum mesothelium and angiogenesis. The purpose of this study was to evaluate the influence of a low-cost drug (doxycycline), with known action on the inhibition of metalloproteinases, in induced peritoneal endometriosis in rats. Thirty adult Wistar rats with endometriosis-induced lesions were divided into three groups: one untreated control group (C, n = 10), one group receiving low dose doxycycline (BD, n = 10) and a group where high dose doxycycline (AD, n = 10) was performed. An evaluation of the lesion area of each rat and immunohistochemical study for primary antibody to matrix metalloproteinase 9 (MMP9) and matrix metalloproteinase inhibitor 2 (TIMP2) was performed. Doxycycline worked by reducing the area of lesions in the BD and AD groups (p = 0.0052) in relation to the C group and reducing the expression of TIMP2 in the AD group (p = 0.0009) in relation to the BD and C groups. There was no significant effect on MMP9 expression in the present study.

Le TIMP-1, inhibiteur naturel des métalloprotéinases matricielles, exerce des effets pléïotropes indépendants de l'inhibition des MMPs et participe au développement de certains cancers et maladies neurodégénératives. Ces effets cytokiniques du TIMP-1 impliquent sa liaison à des récepteurs membranaires dont certains sont caractérisés, la glycoprotéine CD63/intégrine beta 1 et le complexe pro MMP-9/CD44. Cependant les acides aminés ou les domaines du TIMP-1 se liant à ces récepteurs ne sont pas identifiés. Les travaux réalisés au cours de cette thèse mettent en évidence un nouveau récepteur du TIMP-1, la protéine LRP-1. Dans les neurones corticaux murins, le TIMP-1 se fixe aux domaines DII et DIV de LRP-1, est endocyté et induit une réduction de la taille des neurites ainsi qu'une augmentation du volume des cônes de croissance. Afin de caractériser cette interaction, nous avons utilisé une approche originale de modélisation moléculaire associant les analyses de modes normaux et la dynamique moléculaire. Ces analyses in silico ont permis d'identifier un mouvement de pince entre les domaines N et C-terminaux du TIMP-1. Nous avons muté trois résidus (F12, K47 et W105) localisés dans une région essentielle d'un point vue énergétique à l'exécution de ce mouvement. Ces trois mutants n'ont pas d'effet sur la longueur du réseau neuritique et ne sont pas endocytés par LRP-1. En revanche, ils interagissent avec les 2 autres récepteurs (CD63 et proMMP-9) et reproduisent les effets du TIMP-1 sauvage. De plus, nous avons identifié une séquence de 6 acides aminés localisée dans le domaine extracellulaire I de CD63 et essentielle à la liaison avec le TIMP-1. L'ensemble de ces travaux a permis l'identification de régions impliquées dans l'interaction du TIMP-1 avec ses différents récepteurs et pourrait permettre le développement de nouveaux outils pharmacologiques ciblant les activités cytokiniques du TIMP-1. / TIMP-1, a natural inhibitor of matrix metalloproteinases, exerts pleiotropic effects independent of MMP inhibition and thus participates to the development of some cancers and neurodegenerative disorders. These cytokine-like activities require TIMP-1 binding to membrane receptors. Up to date two receptors, CD63/integrin beta 1 and proMMP-9/CD44, have been characterized. Nevertheless, TIMP-1 residues or regions binding these receptors remain unknown. In this work, we have identified the protein LRP-1 as a new receptor for TIMP 1. In mouse cortical neurons, TIMP-1 preferentially binds DII and DIV domains of LRP-1, is internalized via a LRP-1-dependent endocytosis, reduces neurite length and increases growth cone volume. To go deeper into TIMP-1/LRP-1 interaction, we used an original molecular modeling approach which combined normal mode analysis and molecular dynamic. These in silico studies allow us to point out a clamp movement between the N- and C-terminal domains of TIMP-1. Three residues localized in a region that seems essential for the movement have been mutated (F12, K47 and W105) and single mutants have been produced. These mutants do not reduce neurite outgrowth and are not internalized by LRP-1. In contrast, they interact with the two others receptors proMMP-9 and CD63 and induce associated biological effects. Furthermore, we have identified a sequence of six residues localized in the CD63 extracellular domain I and essential for TIMP 1 binding. The set of our data highlighted new regions of TIMP-1 interacting with its receptors and could lead to design novel therapeutic agents targeting the TIMP-1 cytokine like activities.

Submitted by Ana Lúcia Torres (bfmhuap@gmail.com) on 2017-09-29T15:10:17Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação Livia Gusmao.pdf: 1628245 bytes, checksum: 42849e5f87bd042f59660a228ded7164 (MD5) / Approved for entry into archive by Ana Lúcia Torres (bfmhuap@gmail.com) on 2017-09-29T15:10:28Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação Livia Gusmao.pdf: 1628245 bytes, checksum: 42849e5f87bd042f59660a228ded7164 (MD5) / Made available in DSpace on 2017-09-29T15:10:28Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação Livia Gusmao.pdf: 1628245 bytes, checksum: 42849e5f87bd042f59660a228ded7164 (MD5) Previous issue date: 2013 / Hospital Icaraí / O líquen escleroso vulvar está envolvido em uma das vias da carcinogênse da vulva ligada à neoplasia intraepitelial vulvar diferenciada. A metilação da região promotora do DNA é a principal alteração epigenética pela qual um gene é inativado em seres humanos. A metilação do gene P16INK4a, que é um supressor de tumor e atua como inibidor da cinase dependente da ciclina, tem sido descrita como um evento precoce na carcinogênese vulvar. A metilação do gene TIMP-2, que atua como regulador de metaloproteinases, tem sido descrito como marcador de matriz extracelular. Esta pesquisa tem como objetivo estudar a presença da metilação dos genes P16INK4a e TIMP-2 no líquen escleroso vulvar e avaliar a associação das variáveis idade, doença de tireóide, tabagismo, uso de hormônio e prurido vulvar com a metilação dos referidos genes. Trata-se de um estudo transversal, onde foram analisadas 32 amostras obtidas por biópsia de pacientes com líquen escleroso vulvar. As amostras foram submetidas à extração do DNA por meio da técnica do fenol:clorofórmio e à avaliação da metilação dos genes P16INK4a e TIMP-2 pela modificação química do DNA pelo método do bissulfito. O DNA modificado foi submetido à PCR e a visualização do produto pelo gel de poliacrilamida. O estudo da associação de cada uma das variáveis com a metilação de ambos genes não mostrou significado estatístico. Notou-se que 39% (11/28) das amostras exibiram metilação somente para o gene TIMP-2, e nenhuma para o gene P16INK4a isoladamente, enquanto 32% (9/28) apresentaram metilação em ambos genes de forma simultânea. A análise da associação da metilação entre ambos genes mostrou significado estatístico (p=0,0292). Esses resultados sugerem que a associação da metilação entre os genes P16INK4a e TIMP-2 possa promover instabilidade genômica, podendo funcionar como marcador na evolução da doença. Estudos futuros sobre alterações moleculares da matriz extracelular, que por precederem as alterações morfológicas, talvez possam funcionar como sinalizador, individualizando as pacientes com maior risco de evolução do líquen escleroso vulvar para a NIV diferenciada e/ou câncer de vulva / The vulvar lichen sclerosus is involved in one of the pathways of vulvar carcinogenesis linked to differentiated vulvar intraepithelial neoplasia. The methylation of the promoter region of the DNA is the main epigenetic modification in humans in which a gene is inactivated. Methylation of P16INK4a gene, which is a tumor suppressor and acts as an inhibitor of cyclin-dependent kinase, has been described as an early event in carcinogenesis vulva. Methylation of TIMP-2 gene, which acts as a regulator of metalloproteinases, has been described as a marker of extracellular matrix. This research aims to study the presence of methylation of the P16INK4a gene and TIMP-2 in vulvar lichen sclerosus and evaluate the association of age, thyroid disease, smoking, hormone use and vulvar itching with methylation of these genes. It is a cross-sectional study, which analyzed 32 samples obtained by biopsy from patients with vulvar lichen sclerosus. The samples were subjected to DNA extraction by using the technique of phenol: chloroform and evaluation of methylation of the p16INK4a gene and TIMP-2 by chemical modification of DNA by the method of bisulfite. The modified DNA was subjected to PCR and visualization of the product by polyacrylamide gel electrophoresis. The association of each variable in the methylation of both genes showed no statistical significance. It was noted that 39% (11/28) samples showed methylation only to TIMP-2 gene, and none only for the P16INK4a gene, while 32% (9/28) exhibited methylation on both genes simultaneously. The analysis of the association between methylation of both genes showed statistical significance (p = 0.0292). These results suggest that the association between methylation of the P16INK4a gene and TIMP-2 can promote genomic instability, which can act as a marker in the evolution of the disease. Future studies on the molecular alterations of the extracellular matrix, which precede morphological changes, maybe they can function as a signal, separating the patients with higher risk of evolution of vulvar lichen sclerosus to differentiated VIN and / or cancer of the vulva

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / As metaloproteinases-2 (MMP-2) e -14 (MMP-14) e o inibidor tecidual de metaloproteinases tipo 2 (TIMP-2) participam de modo fundamental na transiÃÃo epitelial-mesenquimal e progressÃo tumoral-linfonodal de muitos tipos de cÃncer, inclusive o gÃstrico. O objetivo deste trabalho à avaliar a expressÃo das trÃs enzimas no carcinoma gÃstrico e metÃstases linfonodais e suas possÃveis participaÃÃes na progressÃo tumoral. Foram utilizados 83 casos de gastrectomias por cÃncer gÃstrico (histotipo intestinal = 53 casos; difuso = 30 casos), e seus respectivos linfonodos, dos arquivos do Departamento de Patologia e Medicina Legal/UFC. Foi realizado tissue microarray e imunohistoquÃmica com anticorpo monoclonal anti-MMP-2, anti-MMP-14 e anti-TIMP-2, avaliada atravÃs dos seguintes escores: 0 = ausÃncia de imunomarcaÃÃo ou raras cÃlulas marcadas (< 5%); 1 = marcaÃÃo discreta na maioria (> 50%) das cÃlulas tumorais ou inflamatÃrias mononucleadas (muitos dos quais identificados como macrÃfagos pelo CD68) ou marcaÃÃo moderada em minoria de cÃlulas (< 50%); 2 = marcaÃÃo moderada na maioria (> 50%) das cÃlulas tumorais ou inflamatÃrias mononucleadas ou marcaÃÃo intensa em minoria de cÃlulas (< 50%); 3 = marcaÃÃo intensa na maioria (> 50%) das cÃlulas tumorais ou inflamatÃrias mononucleadas. A expressÃo de MMP-2, MMP-14 e TIMP-2 nos mononucleares associados a tumores ocorreu com maior frequÃncia comparada à imunomarcaÃÃo em mononucleares da mucosa normal, com diferenÃa significativa em relaÃÃo a TIMP-2 (40/53 vs 12/26; *p = 0,0128, teste exato de Fisher). MMP-2 foi muito mais presente nas mulheres (p = 0,0248) enquanto TIMP-2 ocorreu predominantemente apÃs os 50 anos (p = 0,0034). A expressÃo dos trÃs biomarcadores nos carcinomas gÃstricos primÃrios foi muito superior nos mononucleares, em relaÃÃo Ãs cÃlulas neoplÃsicas, sobretudo para a MMP-2 (16/46 vs 5/46; *p = 0,0118), que tambÃm prevaleceu em mononucleares das metÃstases linfonodais em tumores dos histotipos intestinal e difuso (13/16 vs 4/19; ***p = 0,0006). Neste estudo, a expressÃo preponderante dos trÃs imunomarcadores pelos mononucleares do conjuntivo reforÃa o papel central destas cÃlulas e do microambiente tumoral na progressÃo do cÃncer gÃstrico. A maior expressÃo de TIMP-2 no sÃtio primÃrio à sugestiva do efeito inibitÃrio desta enzima sobre MMP-2 e MMP-14, que parecem participar principalmente em fases mais avanÃadas da progressÃo tumoral-linfonodal. A MMP-14, atravÃs dos mononucleares, parece estar mais envolvida na progressÃo do cÃncer gÃstrico difuso do que a MMP-2 e seu inibidor tissular.

Les péri-implantites se caractérisent par une destruction inflammatoire irréversible des tissus qui entourent l’implant dentaire. Leur pathogénie est proche des parodontites, maladies inflammatoires chroniques d’étiologie bactérienne entrainant une destruction irréversible de l’os alvéolaire soutenant les dents. Cependant, la réponse inflammatoire et la résorption osseuse diffèrent entre péri-implantites et parodontites. De plus, les traitements conventionnels non chirurgicaux ne sont pas efficaces pour le traitement des péri-implantites. L’objectif de notre travail est d’analyser les différences entre les médiateurs inflammatoires et cataboliques des tissus péri-implantaires en comparaison aux tissus parodontaux malades après une thérapie conventionnelle non chirurgicale. Nous avons bâti une collection biologique d’explants humains de tissus conjonctifs péri-implantaires ou parodontaux malades. Une analyse Multiplex des milieux conditionnés et histologiques des explants a révélé une expression accrue de l’inhibiteur de métalloprotéase TIMP-2, de l’interleukine IL-10 et du RANK-L dans les tissus péri-implantaires. En accord avec une revue de littérature que nous avons mené dans la seconde partie de notre travail, notre hypothèse est que ces différences s’expliqueraient, tout du moins en partie, par la présence de particules de titane au niveau des tissus péri-implantaires. Notre travail participe à la compréhension de la pathogenèse de la maladie et concourt à l’identification de biomarqueurs potentiels pour le pronostic ou le développement d’approches thérapeutiques. / Peri-implantitis lesions present an irreversible destruction of the peri-implant surrounding tissues. The pathogenic pathway of peri-implantitis is close from the one of periodontitis which is a chronic inflammatory disease caused by a bacterial biofilm leading to irreversible supportive bone destruction around teeth. Still, the inflammatory process and the bone destruction differ between the two diseases. In addition, conventional non-surgical treatments are not effective for the treatment of peri-implantitis. The aim of our work is to analyze the differences between inflammatory and catabolic mediators of peri-implant tissues compared to diseased periodontal tissues after conventional non-surgical therapy. We have constructed a biological collection of human explants of peri-implant or periodontal connective tissue. Multiplex assays of the conditioned media and histological analysis of the explants revealed an increased expression of the TIMP-2 metalloprotease inhibitor, interleukin IL-10, and RANK-L in the peri-implant tissues. In agreement with a review of literature that we conducted in our work, our hypothesis is that these differences can be explained, at least partly, by the presence of titanium wear particles in the peri-implant tissues. Our work contributes to the understanding of the pathogenesis of the disease and contributes to the identification of biomarkers.

No description available.

This dissertation elucidates several independent molecular mechanisms that function in astrocytes and glial tumor cells, and suggest that developmental and inflammatory signals may contribute to the development of brain tumors. First, we analyzed the mechanism of TIMP-1 activation in astrocytes and glioblastoma cells. TIMP-1 expression is activated by IL-1, which is the major neuroinflammatory cytokine, via simultaneous activation of IKK/NF-kB and MEK3/6/p38/ATF-2 pathways in primary human astrocytes. In contrast to astrocytes, TIMP-1 is expressed at lower levels in glioblastomas, and is not regulated by IL-1 due to either dysfunctional IKK/NF-kB or MEK3/6/p38/ATF-2 activation. Thus, we propose a novel mechanism of TIMP-1 regulation, which ensures an increased supply of the inhibitor after tissue injury to limit the ECM degradation. This mechanism does not operate in gliomas, and may in part explain the increased invasiveness of glioma cells.Inflammation has been associated with the development of several cancers, including glioblastoma multiforme. However, it has not been linked to other brain tumors. Here we show for the first time that inflammation is associated with oligodendroglioma tumors as pro-inflammatory cytokines, such as OSM, IL-6, MCP1, MIP1α, and MIP1β and inflammatory markers, such as ACT and COX-2, were expressed at higher levels in oligodendroglioma samples. In addition, cytokine-induced STAT3 signaling, but not NF-kB, is highly activated in the oligodendroglioma patients. Moreover, OSM promotes oligodendroglioma cell proliferation in vitro, and this effect is mediated through STAT3. In summary, oligodendroglioma tumors secrete and respond to inflammatory mediators, with OSM being the major cytokine that activates STAT3 to promote the growth of tumor cells, and express ACT and COX-2 as a hallmark of ongoing inflammation. Since STAT3 promotes the growth of oligodendroglioma, as well as glioblastoma cells, and also regulates gliogenesis, we studied molecular mechanisms of this process in an in vitro differentiation model. We turn our attention to the NFI family of transcription factors since they have recently emerged as novel regulators of the development of vertebral neocortex. We developed a stem cell-neural progenitor-astrocyte differentiation model, in which the generated astrocytes were characterized by proper morphology, increased glutamate uptake, and expression of early and late astrocyte markers. Moreover, we found that NFI-X and NFI-C but not NFI-A or NFI-B, control the expression of GFAP and SPARCL1, the markers of terminal differentiation of astrocytes.In summary, the three mechanisms of gene regulation we studied, provided new insights into astrocyte biology, with the important implications for understanding the basis leading to the development and progression of brain tumors.

No description available.

Abstract Endometrial carcinoma is the most common gynegologic malignancy in developed countries. Due to early symptoms, including abnormal uterine bleeding, endometrial cancer is often diagnosed at an early stage and in that case usually has a good prognosis and high cure rates. However, the nature of the disease is heterogeneous. During the last decades, the improvement in survival rates among endometrial cancer patients has not been significant, suggesting that the traditional clinicopathological factors may be inadequate to identify patients with high-risk disease. Furthermore, aggressive adjuvant treatments can be costly and very toxic. Therefore, better prognostic markers associated with biological aggressiveness of endometrial carcinoma are needed to identify the patients with high-risk disease, and to be able to select the treatment more individually. Gelatinases (MMP-2 and MMP-9) and their tissue inhibitors (TIMP-1 and TIMP-2) have been found to play a role in tumor progression. In the present work, the expression and prognostic value of MMP-2, MMP-9, TIMP-1 and TIMP-2 were assessed in endometrial carcinoma. The patient material consisted of a total of 266 women diagnosed with primary endometrial carcinoma. The tissue expression of immunoreactive proteins was examined in paraffin-embedded tumor sections by immunohistochemical staining using specific antibodies, and the pretreatment serum levels of the proteins were quantitatively measured by ELISA. Tissue MMP-2 expression associated with a worsened prognosis, whereas tissue TIMP-2 overexpression was an indicator of a favorable outcome. Furthermore, we observed a combination of strong MMP-2 and weak TIMP-2 tissue expression to identify a group of women at high risk of adverse outcome in endometrial carcinoma. Patients with negative MMP-2 immunostaining had the best prognosis, regardless of TIMP-2 staining result. In serum measurements, high preoperative TIMP-1 concentration was a prognostic indicator of unfavorable outcome. These results indicate that tissue MMP-2 and TIMP-2 as well as circulating TIMP-1 may be prognostic markers in endometrial carcinoma. Of these, tissue MMP-2 seems to be the most potent prognostic marker. Studies with larger patient materials are needed to further explore the value of these enzymes in clinical practice in endometrial cancer. / Tiivistelmä Kohdunrungon syöpä on yleisin gynekologinen maligniteetti kehittyneissä maissa. Varhaisten oireiden, kuten poikkeavan verisen vuodon, vuoksi kohdunrungon syöpä havaitaan usein varhaisessa vaiheessa, jolloin sen ennuste on hyvä. Taudin käyttäytyminen voi kuitenkin olla moninaista. Viime vuosikymmenten aikana kohdunrungon syöpään sairastuneiden ennuste ei ole merkittävästi parantunut. Vaikuttaisi siltä, että perinteiset ennustetekijät eivät ole riittävän tarkkoja ennustamaan syövän taudinkulkua. Lisäksi liitännäishoidot voivat olla kalliita, ja niihin voi liittyä vakavia haittavaikutuksia. Uusien biologisten ennustetekijöiden löytäminen olisi tärkeää, jotta aggressiivista syöpätyyppiä sairastavat potilaat pystyttäisiin tunnistamaan entistä paremmin, ja hoito kyettäisiin räätälöimään yksilöllisemmin taudinkuvaa vastaavasti. Gelatinaasien (MMP-2 ja MMP-9) sekä niiden kudosinhibiittoreiden (TIMP-1 ja TIMP-2) on havaittu osallistuvan syövän etenemiseen. Tässä tutkimuksessa tarkasteltiin MMP-2:n ja MMP-9:n sekä niiden kudosinhibiittoreiden TIMP-1:n ja TIMP-2:n ilmentymistä ja ennusteellista merkitystä kohdunrungon syövässä. Aineisto käsitti yhteensä 266 primaariseen kohdunrungon syöpään sairastunutta naista. Määritysmenetelminä käytettiin sekä immunohistokemiallista värjäystä parafiiniin valettujen kudosnäytteiden osalta että ELISA-määrityksiä ennen hoitoa otettujen seeruminäytteiden osalta. Syöpäkudoksen runsas MMP-2 -proteiinin ilmentyminen liittyi epäsuotuisaan ennusteeseen, kun taas kasvainkudoksen voimakas TIMP-2 -proteiinin ilmentyminen oli hyvän ennusteen merkki. Lisäksi kasvainkudoksen voimakkaan MMP-2- ja heikon TIMP-2 -proteiinien ilmentymisen yhdistelmän havaittiin liittyvän suurempaan syövästä johtuvaan kuolleisuuteen. MMP-2 -negatiivisten potilaiden eloonjäämisennuste oli paras, TIMP-2 -värjäystuloksesta riippumatta. Seerumin korkea TIMP-1 -pitoisuus oli merkittävä huonontuneen ennusteen merkki. Tutkimuksen tulokset viittaavat siihen, että kasvainkudoksessa esiintyvät MMP-2- ja TIMP-2 -proteiinit samoin kuin seerumin TIMP-1 -pitoisuus voivat ennustaa kohdunrungon syövän kliinistä käyttäytymistä. Kasvainkudoksessa esiintyvä MMP-2 -proteiini vaikuttaisi olevan merkittävin ennusteellinen tekijä, mutta tulosten varmistamiseksi tarvitaan lisää tutkimuksia suuremmilla potilasaineistoilla.