Made available in DSpace on 2015-07-22T20:49:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-11-24. Added 1 bitstream(s) on 2015-08-11T03:26:29Z : No. of bitstreams: 1 Publico-393.pdf: 1637068 bytes, checksum: 4fe3757007f6a049eac930eb08b63c88 (MD5) / O melanoma é o tipo de câncer de pele menos frequente, mas que tem um grande poder de letalidade devido ao seu potencial de formar metástases. Para as células adquirirem a capacidade de formar metástases, estas precisam ter a característica de sobreviver independente de interações com a matriz extracelular e consequentemente apresentar resistência ao anoikis. Por isso, a importância de se estudar as alterações que ocorrem com células tumorais que adquirem essa capacidade. Em nosso laboratório foi desenvolvido um modelo que nos permite estudar diferentes etapas da gênese do melanoma. Melanócitos murinos melan-a que sobreviveram depois de 1, 2, 3 e 4 ciclos de impedimento de ancoragem por 96 horas apresentaram modificações na morfologia e crescimento independente de PMA, e foram denominadas 1C, 2C, 3C e 4C, respectivamente. Diferentes linhagens de melanoma (4C11-, 4C11+, Tm1, Tm5, etc) foram estabelecidas após submeter os esferóides sobreviventes da 4C à diluição limitante. Dados prévios de nosso laboratório mostraram aumento da expressão de Timp1 ao longo da transformação maligna de melanócitos e aumento da resistência ao anoikis. Melanócitos melan-a superexpressando o gene Timp1 adquirem fenótipo de resistência ao anoikis. No entanto, o mecanismo pelo qual Timp1 medeia essa sinalização de sobrevivência não é conhecido. Dados da literatura mostram interação entre CD63, Timp1 e 1-integrinas em células epiteliais de mama humana e que essa interação regula processos fisiológicos como apoptose. Além disso, a glicosilação aberrante em moléculas de adesão celular, como integrinas, pode conferir às células capacidade de sobreviver em condições independentes de ancoragem. O objetivo do presente estudo foi analisar a possível interação entre CD63, Timp1 e 1-integrinas ao longo da transformação maligna de melanócitos, a presença de N-glicosilação aberrante em β1-integrinas e o impacto da N-glicosilação aberrante na resistência ao anoikis. Observou-se interação entre CD63 e Timp1 e CD63 e 1-integrinas nas linhagens 4C, 4C11- e 4C11+, estabelecidas após ciclos de impedimento de ancoragem, já a interação entre Timp1 e 1-integrinas foi observada somente nas linhagens de melanoma 4C11- e 4C11+. A expressão de 1-integrinas na superfície celular está aumentada na linhagem de melanoma agressivo 4C11+, assim como a expressão de Mgat-V e N-glicosilação aberrante. Além disso, o perfil eletroforético da 1-integrina sugere que a mesma apresenta aumento de N-glicosilação aberrante na linhagem de melanoma metastático 4C11+. O tratamento de células de melanoma 4C11+ com o inibidor de N-glicosilação swainsonine resulta em menor capacidade destas células em resistir ao anoikis. Este parece ser o primeiro estudo descrevendo a interação entre Timp1, CD63 e 1-integrinas em células tumorais. Assim, o presente trabalho favorece o entendimento de como Timp1 regula resistência ao anoikis ao longo da transformação maligna de melanócitos. / Although malignant melanoma is the less frequently diagnosed skin cancer, it shows a poor prognosis due its chemoresistance and metastasis development. One of the adquired abilities of transformed cells is anoikis resistance and this property is closely related to metastasis formation. In our laboratory, we developed a model that allows us to study different steps of melanocyte malignant transformation. Melan-a melanocytes surviving after 1, 2, 3 and 4 deadhesion cycles showed modified morphology and independent PMA growth and have been named, 1C, 2C, 3C and 4C cells, respectively. Different melanoma cell lines were established after submitting 4C spheroids to limiting dilution. Previous results of our group showed increased expression of Timp1 along melanoma genesis and its correlation with anoikis resistance. However, the mechanism involved in this signaling is unknown. Published data demonstrated interaction between CD63, Timp1 and 1-integrins in human breast epithelial cells and its role in apoptosis. Furthermore, aberrant glycosylation in cell adhesion molecules such as integrins provides to cells the ability to survive under anchorage-independent conditions. The aim of this work was analyze the possible interaction among CD63, Timp1 and 1-integrins along melanocyte malignant transformation, possible aberrant N-glycosylation patterns of β1-integrins and their impact in anoikis resistance. Aberrant N-glycosylation patterns were observed in tumorigenic cells. We observed interaction between CD63 and Timp1 and between CD63 and 1-integrins in the melan-a-derived cells 4C, 4C11, and 4C11 +, and interaction between Timp1 and 1-integrins only in melanoma cell lines 4C11 - and 4C11 +. The presence of Timp1 in supernatant from 4C11+ conferred to melan-a cells anoikis resistance. The expression of 1-integrins in our study model is increased in aggressive melanoma lineage, 4C11+, as well as the expression of Mgat-V and aberrant N-glycosylation on cell surface. Moreover, the electrophoretic profile of  1-integrin suggests that melanoma metastatic 4C11+. Lineage present increased aberrant N-glycosylation in this molecule. Treatment of melanoma cells 4C11 + with the N-glycosylation inhibitor, swainsonine, resulted in reduced capacity of these cells to resist to anoikis. This seems to be the first study describing the interaction between Timp1, CD63 and 1-integrin in tumor cells and may contribute to a better understanding of how Timp1 regulates resistance to anoikis during the melanocyte malignant transformation. / TEDE / BV UNIFESP: Teses e dissertações

O conceito de campo de cancerização, em dermatologia, sugere que a pele fotodanificada tem maior potencial para o desenvolvimento de neoplasias cutâneas. A terapia fotodinâmica (TFD) é um método não invasivo para o tratamento de queratoses actínicas (QA) múltiplas, possibilitando a abordagem de todo o campo. Vinte e seis pacientes com múltiplas QAs na face foram submetidos a três sessões de TFD com metilaminolevulinato 16% (MAL) e luz vermelha, com intervalo de um mês. Biópsias foram realizadas antes e após três meses da última sessão e o material corado para hematoxilina-eosina e Weigert. O estudo imunohistoquímico foi feito para os marcadores: TP-53, pró-colageno I, Metaloproteinase-1 e Tenascina-C. A avaliação do fotoenvelhecimento global melhorou consideravelmente (p < 0,001) e a cura clínica das QAs foi de 89,5% ao final do estudo. Duas sessões mostraram ser equivalentes a três sessões de TFD. Diminuição significante do grau e extensão da atipia celular (p < 0,001), aumento das fibras colágenas (p = 0,001) e melhora do grau de elastose (p = 0,002) foram observadas. O estudo imunohistoquímico mostrou diminuição da expressão da TP-53 (p = 0,580), aumento de pró-colágeno I (p = 0,477) e de MMP-1 (p = 0,08), embora não houvesse diferença estatisticamente significante. Aumento significativo foi observado para Tenascina-C (p = 0,024). Múltiplas sessões de TFD com MAL induziram melhora clínica e histológica do campo de cancerização. A diminuição da severidade e extensão da atipia celular associada à menor expressão de TP-53 sugerem redução do potencial carcinogênico do campo / The field cancerization concept suggests that photodamaged skin has an increased risk for the development of malignant lesions. Topical photodynamic therapy (PDT) is a non-invasive therapeutic method for multiple actinic keratosis (AK), allowing the possibility of treating the entire surface. Twenty-six patients with photodamaged skin and multiple AKs on the face were submitted to three consecutive sessions of PDT with mehtylaminolevulinate 16% (MAL) and red light, one month apart. Biopsies were performed before and three months after the last treatment session, and stained for hematoxilin-eosin and Weigert. Immunohistochemestry study was performed for TP-53, pro-collagen I, Metalloproteinase-1 and Tenascin-C. The global score for photodamage improved considerably in all patients (p < 0.001). The AK clearance rate was 89.5% at the end of the study. Two treatments were similar to three MAL-PDT sessions. A significant decrease in keratinocytes atypia grade and amount was observed (p < 0.001). A significant increase in collagen deposition (p = 0.001) and improvement of solar elastosis (p = 0.002) were noticed. Immunohistochemical study showed decreased TP-53 expression although not statistically significant (p = 0.580), increased pro-collagen I and MMP-1 expressions (p = 0.477 and p = 0.08) again not statistically significant and a increased expression of Tenascin-C (p = 0.024), which was statistically significant. In conclusion, multiple sessions of MAL-PDT induced clinical and histological improvement of field cancerization. The decrease in severity and extension of keratinocytes atypia associated with a decreased expression of TP-53 suggest a reduced carcinogenic potential of the altered field

Abstract Lymphomas are a heterogeneous group of malignancies, which usually have a good prognosis and high cure rates. Lymphomas are sensitive to chemotherapy and radiotherapy, and many patients can be cured even after a relapse, resulting in a need for effective follow-up. However, the cost-benefit ratio of radiological imaging in predicting the forthcoming relapses is poor. Consequently, there is a need for biological prognostic and predictive markers to distinguish patients at the highest risk of relapse at the time of diagnosis or during follow-up. Despite rapid progress in lymphoma treatments, some patients still die from lymphoma. Thus, more data on the basic biological features of lymphomas are also needed. Gelatinases (MMP-2 and MMP-9) and their tissue inhibitors (TIMP-1 and TIMP-2) have been found to play a role in the progression of solid tumours. TP53 is a tumour suppressor gene, the mutations and protein over-expression of which have been demonstrated to be associated with survival in most cancer types. There is also some evidence that these proteins could have prognostic significance in lymphomas as well. In the present study, the tissue expression, plasma concentrations and clinical value of gelatinases and their tissue inhibitors were evaluated in lymphomas. 249 primary tissue samples from patients with Hodgkin, follicular, or diffuse large B-cell lymphoma were analysed for expression of gelatinases and/or their inhibitors using immunohistochemistry. In follicular lymphoma, p53 protein expression was also investigated. The plasma samples of 126 lymphoma patients and a control group of 44 healthy volunteers were collected and studied by ELISA. TIMP-1 expression correlated with bulky tumour and nodular sclerosis subtype of Hodgkin lymphoma. In follicular lymphoma, p53 over-expression was an independent adverse prognostic factor for survival and a predictor of histological transformation. Plasma MMP-2-TIMP-2 complex appeared to be a potential follow-up marker predicting the risk of relapse in lymphoma patients. Plasma levels of the MMP-2-TIMP-2 complex, proMMP-2, TIMP-2 and proMMP-2/TIMP-2 ratio were at abnormal levels both in patients with newly diagnosed lymphoma and those in remission compared to healthy controls. The clinical significance of these markers needs further studies.

O conceito de campo de cancerização, em dermatologia, sugere que a pele fotodanificada tem maior potencial para o desenvolvimento de neoplasias cutâneas. A terapia fotodinâmica (TFD) é um método não invasivo para o tratamento de queratoses actínicas (QA) múltiplas, possibilitando a abordagem de todo o campo. Vinte e seis pacientes com múltiplas QAs na face foram submetidos a três sessões de TFD com metilaminolevulinato 16% (MAL) e luz vermelha, com intervalo de um mês. Biópsias foram realizadas antes e após três meses da última sessão e o material corado para hematoxilina-eosina e Weigert. O estudo imunohistoquímico foi feito para os marcadores: TP-53, pró-colageno I, Metaloproteinase-1 e Tenascina-C. A avaliação do fotoenvelhecimento global melhorou consideravelmente (p < 0,001) e a cura clínica das QAs foi de 89,5% ao final do estudo. Duas sessões mostraram ser equivalentes a três sessões de TFD. Diminuição significante do grau e extensão da atipia celular (p < 0,001), aumento das fibras colágenas (p = 0,001) e melhora do grau de elastose (p = 0,002) foram observadas. O estudo imunohistoquímico mostrou diminuição da expressão da TP-53 (p = 0,580), aumento de pró-colágeno I (p = 0,477) e de MMP-1 (p = 0,08), embora não houvesse diferença estatisticamente significante. Aumento significativo foi observado para Tenascina-C (p = 0,024). Múltiplas sessões de TFD com MAL induziram melhora clínica e histológica do campo de cancerização. A diminuição da severidade e extensão da atipia celular associada à menor expressão de TP-53 sugerem redução do potencial carcinogênico do campo / The field cancerization concept suggests that photodamaged skin has an increased risk for the development of malignant lesions. Topical photodynamic therapy (PDT) is a non-invasive therapeutic method for multiple actinic keratosis (AK), allowing the possibility of treating the entire surface. Twenty-six patients with photodamaged skin and multiple AKs on the face were submitted to three consecutive sessions of PDT with mehtylaminolevulinate 16% (MAL) and red light, one month apart. Biopsies were performed before and three months after the last treatment session, and stained for hematoxilin-eosin and Weigert. Immunohistochemestry study was performed for TP-53, pro-collagen I, Metalloproteinase-1 and Tenascin-C. The global score for photodamage improved considerably in all patients (p < 0.001). The AK clearance rate was 89.5% at the end of the study. Two treatments were similar to three MAL-PDT sessions. A significant decrease in keratinocytes atypia grade and amount was observed (p < 0.001). A significant increase in collagen deposition (p = 0.001) and improvement of solar elastosis (p = 0.002) were noticed. Immunohistochemical study showed decreased TP-53 expression although not statistically significant (p = 0.580), increased pro-collagen I and MMP-1 expressions (p = 0.477 and p = 0.08) again not statistically significant and a increased expression of Tenascin-C (p = 0.024), which was statistically significant. In conclusion, multiple sessions of MAL-PDT induced clinical and histological improvement of field cancerization. The decrease in severity and extension of keratinocytes atypia associated with a decreased expression of TP-53 suggest a reduced carcinogenic potential of the altered field

Cardiovascular disease (CVD) is ranked the second leading cause of death in Canada, with 53,704 heart disease-related deaths documented in 2020 alone. After a patient sustains cardiac injury, such as a myocardial infarction (MI), the heart is often unable to undergo sufficient self-recovery for healthy cardiac regeneration and repair; this is largely attributed to fibrotic tissue development at the injury site and subsequent pathological ventricular remodeling. The prevalence of MI events has created a considerable demand to develop novel strategies for effective and safe post-MI therapies. Research has indicated that post-MI modifications interfere with endogenous cardiac repair mechanisms, resulting in a pathological state. After an infarction, there is an accumulation of methylglyoxal (MG) at the site of injury. It has been suggested that MG contributes to ventricular fibrotic development, however its underlying mechanism remains unclear. Additionally, the effects that the post-MI cardiac environment, specifically MG accumulation, has on post-MI therapies and biomaterials has not been sufficiently established. Accordingly, the primary focus of this research project is to elucidate the effects of MG on the collagen-rich extracellular matrix (ECM) of the heart and key cardiac cells involved in the repair process. Further, the interaction between MG and a promising collagen-based hydrogel therapy is investigated, exploring the effects of MG on the hydrogel’s degradative process. It was found that the MG modification of hydrogels did not alter the degradation rate. Additionally, the degradation products of hydrogels, and MG-modified substrates did not affect the properties and formation of myofibroblasts.

Made available in DSpace on 2015-07-22T20:50:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-04-29. Added 1 bitstream(s) on 2015-08-11T03:25:24Z : No. of bitstreams: 1 Publico-00211a.pdf: 1874263 bytes, checksum: 9fa88ecbdc678f5d003c91ff7ce39eca (MD5). Added 1 bitstream(s) on 2015-08-11T03:25:25Z : No. of bitstreams: 2 Publico-00211a.pdf: 1874263 bytes, checksum: 9fa88ecbdc678f5d003c91ff7ce39eca (MD5) Publico-00211b.pdf: 1651291 bytes, checksum: 797f8caf18af70f6f0cf21a1510830fa (MD5). Added 1 bitstream(s) on 2015-08-11T03:25:25Z : No. of bitstreams: 3 Publico-00211a.pdf: 1874263 bytes, checksum: 9fa88ecbdc678f5d003c91ff7ce39eca (MD5) Publico-00211b.pdf: 1651291 bytes, checksum: 797f8caf18af70f6f0cf21a1510830fa (MD5) Publico-00211c.pdf: 1136167 bytes, checksum: b8d06cb6386ddbf8bce0ebd5b67f3d7a (MD5) / Objetivo: Estudar a imunoexpressão das proteínas metaloproteinase-1, metaloproteinase-7 e p53 em pacientes portadores de adenocarcinoma colorretal, correlacionando com os fatores prognósticos clínico-patológicos. Métodos: Foram analisados tecidos fixados em formol e dispostos em blocos de parafina dos tumores de 82 pacientes, por imunohistoquímica, pelo método da estreptavidina-biotina, usando-se a técnica de arranjo em matriz de amostras teciduais (tissue microarray). Na avaliação da positividade dos marcadores foi utilizado um escore categórico, que predeterminou o valor de corte na percentagem de células coradas do tumor. As expressões teciduais das proteínas foram correlacionadas com as varáveis representadas pelo grau de diferenciação celular, estadiamento, tempo livre de doença, recidiva, sobrevida e mortalidade específica. Foram empregados os testes do qui-quadrado e de Kaplan-Meier para verificar as associações dos marcadores com as varáveis estudadas. Para testar a significância das diferenças entre as curvas do tempo livre de doença e da sobrevida foram utilizados os testes de Log-Rank e Wilcoxon. Resultados: A metaloproteinase-1 foi positiva em todos os tumores. A metaloproteinase-7 foi positiva em 50 (61%) e negativa em 32 (39%) tumores. A p53 foi positiva em 70 (85,4%) e negativa em 12 (14,6%) tumores. A correlação da imunoexpressão dos marcadores realizada separadamente e em conjunto não apresentou associação comsignificância estatística com nenhuma das variáveis analisadas. Conclusão: Não houve correlação da associação entre a imunoexpressão das proteínas metaloproteinase-1, metaloproteinase-7 e p53 com a recidiva tumoral, mortalidade, intervalo de tempo livre de doença, sobrevida, grau de diferenciação celular e estadiamento do câncer colorretal. / Purpose: To analyse the immunoexpression of metalloproteinase-1, metalloproteinase-7 and also the p53 in colorectal adenocarcinoma and correlate it with the clinical pathological prognostic factors. Methods: Formalin-fixed, wax-embedded tissue of the tumours of 82 patients were analysed by immunohystochemistry through the streptavidin-biotin method, using the tissue microarray technique. When evaluating the positivism of the markers a categorical score was used, which predetermined the cutting value on the percentage of the tumour’s cored cells. The protein’s tissue expressions were correlated with the variation represented by the degree of cellular differential, stage, relapse-free survival, recurrence, survival and specific mortality. The tests of the qui-square and Kaplan-Meyer were applied with the aim to verify the markers association with the studied variations. In order to test the significance of the differences between the curves of relapse-free survival and of the overall survival, the tests of Log-Rank and Wilcoxon were utilized. Results: The metalloproteinase-1 was found to be positive in all the tumours. The metalloproteinase-7 was found to be positive in 50 (61%) and negative in 32 (39%) of the tumours. The p53 was found to be positive in 70 (85.4%) and negative in 12 (14.6 %) of the tumours. The correlation of the markers expressions separately and in conjunction produced any significant statistical data. Conclusion: There was no correlation between immunoexpression of metalloproteinase-1, metalloproiteinase-7 and p53 with recurrence, mortality, relapse-free survival, survival, degree of cellular differential and stage in colorectal cancer. / TEDE / BV UNIFESP: Teses e dissertações

Abstract Matrix metalloproteinases (MMPs) are the zinc-dependent endopeptidases which belong to a large family of proteases. MMPs are responsible for degradation and remodeling of extracellular matrix proteins during growth, organogenesis and tissue turnover. Besides their role in physiological processes, MMPs also influence various pathological processes, i.e., inflammatory diseases and cancers, in which MMPs may ultimately lead to unwanted tissue destruction. One of the most widely studied MMPs is MMP-8. MMP-8 was previously thought to be expressed only by neutrophils. Presently, it is evident that MMP-8 can be expressed in a wide range of cells such as macrophages, plasma cells, T-cells, endothelial cells, smooth muscle cells, oral epithelial cells, fibroblasts etc. MMP-8 has been previously studied in inflammation and malignancies. High serum MMP-8 concentration is associated with the presence of atherosclerosis and poor cardiovascular disease prognosis, while higher plasma MMP-8 levels protect against lymph node metastasis. Certain MMP-8 gene variations can alter promoter activity and subsequent gene expression. MMP-8 gene variation influences obstetrical outcomes, and lung and breast cancer prognosis. For our study, we hypothesized that systemic levels of MMP-8 correlate with its genetic variations and appear as novel risk markers for disease. We aimed to address the potential role of MMPs and their regulators with a special focus on MMP-8 in distinct sets of inflammatory and malignant diseases, i.e. arterial diseases, and head and neck squamous cell cancer (HNSCC). We demonstrated that the combination of high serum MMP-8 and low myeloperoxidase (MPO) levels is an important risk marker for arterial disease. Further, we demonstrated that MMP-8 gene variation is protective against arterial diseases. Interestingly, we were able to demonstrate an association between MMP-8 gene variation and serum MMP-8 concentration in a healthy population. On the other hand, we showed that plasma tissue inhibitor of matrix metalloproteinases (TIMP-1) concentration is associated with survival in HNSCC patients and TIMP-1 genotype is associated with plasma TIMP-1 levels in women only. Collectively, our study showed that serum MMP-8 levels can be used as an important risk marker in arterial disease and TIMP-1 levels in HNSCC patients. Based on our results, the hypothesis raised has been widely confirmed. Additionally, our study has warranted the need for further investigation involving a larger number of patients. If our results are replicable, serum MMP-8 and plasma TIMP-1 could be used to develop diagnostic tools as well as treatment regimes in clinics. / Tiivistelmä Matriksin metalloproteinaasit (MMP:t) ovat sinkkiriippuvaisia endopeptidaaseja, jotka kuuluvat laajaan proteiineja pilkkovaan proteolyyttiseen entsyymi perheeseen. MMP:ien tehtävä on pilkkoa ja uudelleen muokata soluväliaineen proteiineja kasvun, elinten kehityksen ja kudosten uusiutumisen aikana, mutta MMP:t toimivat aktiivisesti myös patologisissa prosesseissa, kuten tulehdustiloissa ja syövissä. Syövissä MMP:en vaikutus voi johtaa ei-toivottuun kudostuhoon. Yksi laajimmin tutkituista MMP-ryhmän entsyymeistä on MMP-8, jonka alunpitäen ajateltiin ilmenevän vain neutrofiileissä. Nykytietämyksen mukaan MMP-8:aa ilmentyy myös mm. makrofaageissa, plasmasoluissa, T-soluissa, endoteelisoluissa, sileälihassoluissa, suun limakalvon epiteelisoluissa ja fibroplasteissa. MMP-8:aa on aikaisemmin tutkittu erityisesti tulehdustiloissa ja pahanlaatuisissa kasvaimissa. Korkean MMP-8 seerumipitoisuuden on havaittu liittyvän valtimokovettumatautiin ja huonoon ennusteeseen sydän- ja verisuonisairauksissa, kun taas kohonnut MMP-8:n pitoisuus plasmassa suojaa imusolmuke-etäpesäkkeiltä. Tiedetään, että tietyt muutokset MMP-8:n geenissä voivat muuttaa sen promoottoriaktiviteettia ja täten säädellä geenin ilmentymistä. MMP-8:n geenimuutokset vaikuttanevat raskaudenkulkuun sekä keuhko- ja rintasyövän ennusteeseen. Tutkimushypoteesimme mukaan MMP-8:n seerumipitoisuudet riippuvat vaihtelusta MMP-8:aa koodaavassa geenissä ja niitä voidaan pitää uusina riskinarvioinnin merkkiaineina tautitiloissa. Tavoitteenamme oli osoittaa yleisesti MMP:ien ja erityisesti MMP-8:n sekä näiden proteinaasien säätelytekijöiden merkitys tietyissä tulehdustiloissa ja maligniteeteissa, kuten sepelvaltimotaudissa ja pään ja kaulan alueen syövissä. Havaitsimme, että korkea MMP-8 seerumipitoisuus ja alhainen myeloperoksidaasitaso yhdistyvät vahvasti valtimotautiriskiin. Lisäksi osoitimme, että tietty MMP-8:n geenimuunnos on suojaava tekijä valtimotaudille ja että MMP-8:n seerumikonsentraatio on siitä riippuvainen terveillä tutkituilla. Tämän lisäksi todensimme, että MMP:n kudosestäjän (tissue inhibitor of matrix metalloproteinases, TIMP-1) plasmapitoisuus liittyy pään ja kaulan alueen levyepiteelisyöpää sairastavien potilaiden eloonjääntiin ja että TIMP-1:n genotyyppi liittyy sen plasmapitoisuuteen ainoastaan naisilla. Tulostemme mukaan seerumin MMP-8-pitoisuutta voidaan pitää hyvänä riskinarviointivälineenä verisuonitaudeissa sekä TIMP-1-pitoisuutta vastaavasti pään ja kaulan alueen levyepiteelisyövissä. Saadut tulokset tukevat olettamustamme, jonka mukaan MMP-8 on tärkeä tautimarkkeri. Tämä on lisännyt kiinnostusta selvittää MMP:ien merkitystä laajemmin muissa tulehdustiloissa ja syövissä. Jos tulokset saadaan toistetuksi laajemmassa tutkimusaineistossa, seerumin MMP-8:sta voidaan kehittää kliinisten ja analyyttisten laboratorioiden käyttöön sopiva diagnostinen menetelmä.

INTRODUÇÃO: A HD está presente em aproximadamente 50% dos pacientes com OIV devido HPB e 30% dos casos não apresentarão melhora após o tratamento cirúrgico. Até o momento, nenhuma característica clínica pode predizer acuradamente quais pacientes serão beneficiados. Há espaço para o aprimoramento de novos métodos diagnósticos não invasivos capazes de discriminar quais os melhores candidatos à cirurgia. Neste estudo nós analisamos o papel de cinco biomarcadores urinários moleculares associados à HD e à OIV em pacientes com HPB que serão submetidos à RTUP. MÉTODOS: Um estudo prospectivo e controlado analisou 71 pacientes candidatos à RTUP devido HPB, submetidos ao procedimento cirúrgico entre 2011 a 2016. O grupo controle foi composto por pacientes assintomáticos apresentando IPSS menor que 6, volume prostático menor que 30 gramas, ausência de resíduo pósmiccional e fluxometria máxima >= 15ml/s. Todos os pacientes do grupo de estudo realizaram estudo urodinâmico no pré-operatório e 63 pacientes no período pósoperatório. Nós analisamos a presença, o período de início (primeira vs segunda metade do enchimento vesical) e a amplitude ( 40 cmH2O) das CVIs, assim como o grau de obstrução infravesical. A coleta da urina foi realizada no pré-operatório para os pacientes do grupo de estudo. A urina foi analisada para 5 quimiocinas, citocinas e fatores de crescimento usando teste de ELISA. Os valores de concentração das proteínas foram analisados de forma absoluta e normalizados para os níveis de creatinina (/Cr). RESULTADOS: A idade média dos pacientes foi 67 anos (50 a 88). A HD estava presente em 39 (54,9%) pacientes. De acordo com aferições pré-operatórias, a média da concentração urinária de IL-6/Cr (p=0,007), MCP-1(p=0,000), MCP-1/Cr (p=0,000), EFG/Cr (p=0,044) e MMP-1/Cr (p=0,043) estavam respectivamente cerca de 4,5x, 7,1x, 23x, 1,7x e 2,2x vezes significativamente aumentadas em relação aos controles assintomáticos. O nível de EGF foi 1,3 vezes maior nos pacientes que iniciaram CVI tardiamente quando comparados àqueles que iniciaram as contrações na fase inicial de enchimento vesical (p=0,044). A amplitude das CVIs, o fluxo urinário, a complacência, o índice de contratilidade, e o schafer não apresentaram correlações estatísticas com as proteínas estudadas. Em relação a presença de HD, os níveis urinários de IL-6 (p=0,003), MCP-1(p=0,002), e MCP-1/Cr (p=0,002) foram respectivamente 1,8x, 2x, e 2,7 vezes aumentados em relação aos pacientes submetidos à cirurgia sem HD ao estudo urodinâmico. O MCP-1/Cr foi o marcador com melhores propriedades diagnósticas para HD, apresentando área sob a curva (AUC) de 0,71 (95% CI 0,59 a 0,84). A dosagem do MCP-1 não ajustada pela creatinina apresentou uma AUC de 0,71 (95% CI 0,59 a 0,83). A IL-6, por sua vez, teve uma AUC de 0,70 (95% CI 0,58 a 0,82). Todos os outros biomarcadores apresentaram propriedades inadequadas neste cenário para avaliação da HD e OIV. Em relação à resolução ou persistência da HD após 12 meses de tratamento cirúrgico, os níveis dos biomarcadores NGF/Cr (p=0.005) e MMP-1/Cr (p=0.021) foram superiores entre os pacientes que não obtiveram interrupção das CVIs no pós-operatório. Demonstraram serem úteis para predizer a persistência da HD no pós-operatório, o NGF/Cr com AUC de 0,77 (95% CI 0,62 à 0,92) (p=0,006) e o MMP-1/Cr com AUC de 0,72 (95% CI 0,56 à 0,88) (p=0,022). CONCLUSÕES: Vias neuronais parecem estar relacionadas com o período de início das CVIs durante a fase de enchimento vesical. A presença de níveis elevados na urina de biomarcadores inflamatórios e de reparo tecidual sugere um papel da inflamação na gênese da HD, e pode ajudar no diagnóstico não invasivo deste achado no pré-operatório. MCP-1, MCP-1 /Cr e IL-6 foram associados a presença de de HD ao estudo urodinâmico em pacientes com HPB candidatos a RTUP. O nível de EGF/Cr foi associado a contrações vesicais tardias. O MMP-1/Cr foi relacionado a maior pressão detrusora. O biomarcador MCP-1/Cr demonstrou-se ser útil no diagnóstico de HD nos pacientes com HPB. NGF/Cr e MMP-1/Cr demonstraram-se ser úteis para predizer a persistência da HD no pós-operátorio / INTRODUCTION: Detrusor hyperactivity (DH) is present in approximately 50% of patients with bladder outlet obstruction (BOO) due to benign prostatic hyperplasia (BPH), and 30% of the cases will not show improvement after surgical treatment. To date, no clinical feature can accurately predict which patients will benefit from surgical treatment. This rate can be improved because new noninvasive diagnostic methods are capable of determining the best candidates for surgery. In this study, we analyzed the role of five molecular biomarkers in urine associated with DH and BOO in patients with BPH undergoing transurethral resection of the prostate (TURP). METHODS: This prospective and controlled study analyzed 71 patients who were candidates for TURP due to BPH and underwent surgery between 2011 and 2016. The control group consisted of asymptomatic patients with International Prostate Symptom Scores (IPSSs) less than 6, prostate volume less than 30 grams, absence of estimated postvoid residual volume and a maximum flow rate >= 15 ml/s. All patients in the study group underwent a preoperative urodynamic study, with 63 patients undergoing a repeat urodynamic study during the postoperative period. We analyzed the presence, onset period (first vs second half of bladder filling) and amplitude ( 40 cmH2O) of the involuntary detrusor contractions (IDCs), as well as the degree of BOO. Urine was collected preoperatively from patients in the study group. The urine was analyzed for five chemokines, cytokines and growth factors using ELISAs. The protein concentrations were analyzed as absolute and creatinine (/Cr)-adjusted values. RESULTS: The mean age of the patients was 67 years (50 to 88). DH was present in 39 (54.9%) patients. According to preoperative measurements, the mean urine concentrations of IL-6/Cr (p = 0.007), MCP-1 (p = 0.000), MCP-1/Cr (p = 0.000), EGF/Cr (p = 0.044) and MMP-1/Cr (p = 0.043) were approximately 4.5, 7.1, 23, 1.7 and 2.2 times, respectively, those of asymptomatic controls (all significantly increased). The EGF level was 1.3 times higher in patients with late IDCs than in those whose contractions started in the early stage of bladder filling (p = 0.044). The IDC amplitude, urinary flow, compliance, bladder contractility index, and Schafer\'s grade were not significantly correlated with the proteins studied. In patients with DH, urinary levels of IL-6 (p = 0.003), MCP-1 (p = 0.002), and MCP- 1/Cr (p = 0.002) were 1.8, 2, and 2.7 times higher, respectively, than in patients without DH who underwent surgery, according to the urodynamic study. MCP- 1/Cr had the best diagnostic properties for DH, with an area under the curve (AUC) of 0.71 (95% CI: 0.59 to 0.84). The non-Cr adjusted MCP-1 concentration had an AUC of 0.71 (95% CI: 0.59 to 0.83). Additionally, IL-6 had an AUC of 0.70 (95% CI: 0.58 to 0.82). All other biomarkers were inadequate for DH and BOO evaluation. Regarding the resolution or persistence of DH 12 months after surgery, the NGF/Cr (0.13 vs 0.08, p = 0.005) and MMP-1/Cr (0.11 vs 0.04, p = 0.021) levels were higher in patients for whom the IDCs continued during the postoperative period. The following factors were shown to be useful for predicting the persistence of DH during the postoperative period: NGF/Cr, with an AUC of 0.77 (95% CI: 0.62 to 0.92) (p = 0.006), and MMP-1/Cr, with an AUC of 0.72 (95% CI: 0.56 to 0.88) (p = 0.022). CONCLUSIONS: Neural pathways appear to be related to the onset period of IDCs during bladder filling. The presence of high urinary levels of inflammatory and tissue repair biomarkers suggests a role of inflammation in the genesis of DH and may aid in the noninvasive diagnosis of this condition during the preoperative period. MCP-1, MCP-1/Cr and IL-6 were associated with the presence of DH in the urodynamic studies of patients with BPH who were candidates for TURP. The EGF/Cr level was associated with late detrusor contractions. MMP-1/Cr was associated with increased detrusor pressure. The MCP-1/Cr biomarker was shown to be useful for the diagnosis of DH in patients with BPH. NGF/Cr and MMP-1/Cr were shown to be useful in predicting the persistence of DH during the postoperative period

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.