In this work, a new polyurethane (PU) chain extender was developed to incorporate a Glycine-Leucine (Gly-Leu) dipeptide, the cleavage site of several matrix metalloproteinases. PUs were synthesized with either the Gly-Leu-based chain extender (Gly-Leu PU) or a phenylalanine-based chain extender (Phe PU). Both PUs had high molecular weight averages (Mw > 125,000 g/mol) and were phase segregated, semi-crystalline polymers (Tm ~ 42°C) with a low soft segment glass transition temperature (Tg < -50°C). Uniaxial tensile testing of PU films revealed that the polymers could withstand high ultimate tensile strengths (~ 8-13 MPa) and were flexible with breaking strains of ~ 870-910% but the two PUs exhibited a significant difference in mechanical properties. The Phe and Gly-Leu PUs were electrospun into porous scaffolds for degradation and cell-based studies. Fibrous Phe and Gly-Leu PU scaffolds were formed with randomly organized fibers and an average fiber diameter of approximately 3.6 µm. In addition, the Phe PU was electrospun into scaffolds of varying architecture to investigate how fiber alignment affects the orientation response of cardiac cells. To achieve this, the Phe PU was electrospun into aligned and unaligned scaffolds and the physical, thermal, and mechanical properties of the scaffolds were investigated. The degradation of the Phe and Gly-Leu PU scaffolds was investigated in the presence of active MMP-1, active MMP-9, and a buffer solution over 28 days to test MMP-mediated and passive hydrolysis of the PUs. Mass loss and structural assessment suggested that neither PU experienced significant hydrolysis to observe degradation over the course of the experiment. In cell-based studies, Phe and Gly-Leu PU scaffolds successfully supported a high density of viable and adherent mouse embryonic fibroblasts (MEFs) out to at least 28 days. Culturing murine embryonic stem cell-derived cardiomyocytes (mESCDCs) alone and with MEFs on aligned and unaligned Phe PU scaffolds revealed both architectures supported adherent and functionally contractile cells. Importantly, fiber alignment and coculture with MEFs improved the organization and differentiation of mESCDCs suggesting these two parameters are important for developing engineered myocardial constructs using mESCDCs and PU scaffolds.

Darbo uždaviniai: 1. Nustatyti pradinės AGDD paplitimą vidutinio amžiaus (40 – 64 metų) pacientų, sergančių IŠL grupėje bei atsitiktinėje to paties amžiaus Kauno miesto gyventojų imtyje. 2. Palyginti pacientų, sergančių tik IŠL ir IŠL bei pradinės AGDD klinikinius duomenis. 3. Nustatyti MMP-2 (-735 C/T), MMP-2 (-1306 C/T), MMP-3 (-1171 5A/6A), MMP-9 (-1562 C/T) genotipų dažnį bei genotipų derinių įtaką AGDD susiformavimui. 4. Nustatyti MMP-2 (-735 C/T), MMP-2 (-1306 C/T), MMP-3 (-1171 5A/6A), MMP-9 (-1562 C/T) genotipų dažnį esant minkštoms ir kietoms drūzoms, sergant AGDD. 5. Nustatyti MMP-2 (-735 C/T), MMP-2 (-1306 C/T), MMP-3 (-1171 5A/6A), MMP-9 (-1562 C/T) genotipų dažnį bei genotipų derinių įtaką AGDD ir IŠL drauge bei tik IŠL pasireiškimui. 6. Nustatyti funkcinio kontrastinio jautrumo tyrimo rodmenis pacientams, sergantiems pradine lengva ir pradine vidutine AGDD bei spalvų juslės pokyčius sergantiems pradine AGDD, ir oftalmologiškai sveikiems pacientams. / The goals were as follows: 1. To determine the prevalence of AMD in patients with IHD and compare with the prevalence in a random sample of Kaunas population (at 40-64 yrs old). 2. To compare the main clinical characteristics of the patients exhibiting early AMD and IHD together with the patients with IHD alone. 3. To determine the frequency of the genotypes of the matrix metalloproteinases (MMP-2 (-735 C/T), MMP-2 (-1306 C/T), MMP-3 (-1171 5A/6A), MMP-9 (-1562 C/T)), and genotype combinations that have an influence on the development of early AMD. 4. To determine the frequency of the genotypes of the matrix metallo¬proteinases (MMP-2 (-735 C/T), MMP-2 (-1306 C/T), MMP-3 (-1171 5A/6A), MMP-9 (-1562 C/T)), in early AMD patients with soft or hard drusen. 5. To determine the frequency of the genotypes of the matrix metallo¬proteinases (MMP-2 (-735 C/T), MMP-2 (-1306 C/T), MMP-3 (-1171 5A/6A), MMP-9 (-1562 C/T)), and the influence of genotype combi¬na¬tions on the development of AMD and IHD together, and only on IHD development. 6. To determine the results of functional acuity contrast sensitivity test in patients with early mild and early intermediate AMD, and color contrast sensitivity in patients with AMD, and in ophthalmologically healthy patients.

In the intestinal epithelium, Wnt signaling drives the expression of the genes encoding tyrosine kinase receptors EphB2 and EphB3 and represses the expression of their membrane-tethered ligands, ephrin-Bs. Eph-ephrin interactions result in cellular repulsion and are involved in boundary formation. The project of this thesis is to understand the mechanism by which EphB−ephrin-B signals restrict cell positioning of cell types (cell sorting) in the normal intestinal epithelium and suppress colorectal cancer (CRC) progression beyond the earliest stages. We have demonstrated that at the onset of CRC EphB receptors impair the expansion of tumor cells through a mechanism dependent on E-cadherin–mediated adhesion. We show that EphB-mediated compartmentalization restricts the spreading of EphB+ tumor cells into ephrin-B1+ territories in vitro and in vivo. Our results indicate that CRC cells must silence EphB expression to avoid repulsive interactions imposed by normal ephrin-B1+ intestinal cells at the onset of tumorigenesis. We have discovered that cell sorting is the outcome of two integrated mechanisms: cell contraction/repulsion and differential cell adhesion. The latter is the driving force to induce EphB/ephrin-B−mediated cell compartmentalization. We have developed in vitro models to analyze the mechanisms that induce E-cadherin remodeling upon EphB activation. We found RhoA, p120-catenin and the metalloproteinase ADAM10 as downstream effectors of EphB signaling involved in the control of cell sorting in CRC cells. / A l'epiteli intestinal, la ruta de senyalització Wnt indueix l'expressió dels gens que codifiquen per als receptors tirosina kinasa EphB2 i EphB3 i reprimeixen la dels seus lligands transmembrana, efrines de tipus B. Les interaccions Eph-efrina causen repulsió cel·lular i estan implicades en la formació de fronteres entre compartiments. La finalitat d'aquesta tesi és entendre el mecanisme pel qual la senyalització per EphB−efrina-B restringeix el posicionament dels diferents tipus cel·lulars a l'epiteli intestinal normal i suprimeix la progressió del càncer colorectal (CRC) en els primer estadis. Hem demostrat que, a l’inici del CRC, els receptors EphB restringeixen l'expansió de les cèl·lules tumorals a través d'un mecanisme depenent d'adhesió intercel·lular a través d’E-cadherina. En aquest treball es mostra in vitro i in vivo que la compartimentalització mitjançada per la senyalització dels receptors EphB restringeix l’invasió de les cèl·lules tumorals EphB+ als territoris efrina-B+. Aquests resultats indiquen que les cèl·lules de CRC han de silenciar l’expressió d'EphB per evitar les interaccions repulsives imposades per les cèl·lules intestinals normals efrina-B+ circumdants al començament del procés de tumorigènesi. Hem pogut discernir que el reordenament cel·lular per senyals EphB−efrina-B és el resultat de dos mecanismes integrats: la contracció/repulsió intercel·lular i l’adhesió diferencial entre diferents poblacions cel·lulars. Aquesta última és la força principal que condueix a la compartimentalització cel·lular mitjançada per EphB−efrina-B. Hem desenvolupat models in vitro per analitzar els mecanismes que provoquen el remodelament de la E-cadherina sota la senyalització per EphB. Presentem RhoA, p120-catenina i ADAM10 com a efectors de la senyalització de la ruta EphB implicats en el control de la compartimentalització cel·lular en el CRC.

Thesis (Ph.D.)--University of Alberta, 2010. / A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Doctor of Philosophy, Medicine. Title from pdf file main screen (viewed on April 27, 2010). Includes bibliographical references.

Η σεργλυκίνη (SG) είναι μια πρωτεογλυκάνη που εκφράζεται και εκκρίνεται από το σύνολο σχεδόν των αιμοποιητικών κυττάρων, ενώ αποτελεί την κύρια πρωτεογλυκάνη η οποία εκκρίνεται από τις κυτταρικές σειρές πολλαπλού μυελώματος (ΠΜ). Έχει βρεθεί ότι η SG συμμετέχει στη ρύθμιση πληθώρας παραγόντων που εμπλέκονται σε αντιδράσεις φλεγμονής. Τα αποτελέσματά μας δείχνουν ότι η SG που εκκρίνεται από τα μυελωματικά κύτταρα έχει την ικανότητα να αναστέλλει τόσο την κλασσική όσο και τη λεκτινική οδό του συστήματος του συμπληρώματος, ενώ δε φαίνεται να έχει καμία επίδραση στο εναλλακτικό μονοπάτι. Επιπρόσθετα η SG δεν έχει την ικανότητα να προκαλεί την ενεργοποίηση κάποιου από τα τρία μονοπάτια του συμπληρώματος. Βρέθηκε ότι η ανασταλτική δράση της SG εκδηλώνεται μέσω της αλληλεπίδρασής της με τους παράγοντες C1q και MBL. Οι γλυκοζαμινογλυκανικές (GAGs) αλυσίδες της SG είναι υπεύθυνες για τη δέσμευση με την κολλαγονούχα ουρά του παράγοντα C1q, ενώ στη δέσμευση με την MBL πρωτεΐνη πέρα από τη συμμετοχή των GAG αλυσίδων απαιτείται και ο πρωτεϊνικός κορμός της SG. Επιπλέον βρέθηκε ότι αλυσίδες CS-E ελαττώνουν την ικανότητα δέσμευσης της SG με τα μόρια C1q και MBL. Οι αλληλεπιδράσεις της SG με τον C1q και την MBL βρέθηκε ότι είναι ιοντικού χαρακτήρα και σε αντίθεση με τη δέσμευση της SG με την MBL, που εξαρτάται από την παρουσία των ιόντων Ca2+, η αλληλεπίδραση της SG με τον C1q είναι ανεξάρτητη των ιόντων αυτών. Αν και τα επίπεδα της SG στον ορό των ασθενών με ΠΜ εμφανίζονται αυξημένα σε σχέση με τους φυσιολογικούς μάρτυρες, δεν εντοπίστηκαν στατιστικά σημαντικές διαφορές στην δραστικότητα της κλασσικής και της εναλλακτικής οδού του συμπληρώματος στους ασθενείς με ΠΜ και στους φυσιολογικούς δότες. Ωστόσο ενδιαφέρον αποτελεί το γεγονός ότι στους ασθενείς με ΠΜ τα υψηλά επίπεδα SG στον ορό εμφάνισαν τάση συσχέτισης με ελαττωμένη δραστικότητα της κλασσικής οδού του συμπληρώματος. Επιπρόσθετα, η SG που εκκρίνεται απ τα μυελωματικά πλασματοκύτταρα έχει την ικανότητα να προστατεύει τα κύτταρα αυτά από την επίδραση του συμπληρώματος, μετά την ενεργοποίησή του με τη χρήση φαρμάκων. Φαίνεται ότι και η SG της μεμβράνης των μυελωματικών κυττάρων παρουσιάζει προστατευτική δράση έναντι του συμπληρώματος, μιας και κύτταρα που δεν εκφράζουν SG στην επιφάνειά τους είναι δύο με τρείς φορές περισσότερο ευαίσθητα στη δράση του συμπληρώματος, σε σχέση με κύτταρα που την εκφράζουν. Προτείνουμε ότι τόσο η εκκρινόμενη όσο και η δεσμευμένη στην κυτταρική επιφάνεια SG προστατεύουν τα μυελωματικά πλασματοκύτταρα κατά την ανοσοθεραπεία και συμβάλουν στην επιβίωσή τους. Στην παρούσα μελέτη δείξαμε ότι η SG αλληλεπιδρά με το κολλαγόνο τύπου Ι, την πιο άφθονη μορφή κολλαγόνου στο οστό. Επιπλέον βρέθηκε ότι η SG της κυτταρικής επιφάνειας συμμετέχει στην προσκόλληση των μυελωματικών κυττάρων στο κολλαγόνο τύπου Ι. Η αλληλεπίδραση αυτή φαίνεται ότι επάγει την έκφραση από τα μυελωματικά πλασματοκύτταρα των ΜΜΡ-2 και ΜΜΡ-9. Επιπρόσθετα, υπολογίστηκαν τα επίπεδα των ΜΜΡ-2 και ΜΜΡ-9 στον ορό και στο μυελό ασθενών με ΠΜ. Βρέθηκε ότι στον ορό των ασθενών με ΠΜ τα επίπεδα των δύο ενζύμων εμφανίζονται ελαττωμένα σε σχέση με τα φυσιολογικά δείγματα, ενώ αντίθετα στον μυελό των ασθενών η ΜΜΡ-2 βρέθηκε σημαντικά αυξημένη σε σχέση με τους φυσιολογικούς δότες. Καμία μεταβολή δεν παρατηρήθηκε στα επίπεδα της ΜΜΡ-9. Τέλος τα επίπεδα της ΜΜΡ-2 του μυελού ασθενών με ΠΜ βρέθηκε ότι σχετίζονται τόσο με τα επίπεδα του ενζύμου στον ορό, όσο και με τον δείκτη οστικής απορρόφησης ΝΤx, υποδηλώνοντας τη συμμετοχή της στην παθοβιοχημεία της οστικής νόσου που εμφανίζεται στο ΠΜ. Αντιθέτως καμία συσχέτιση δεν βρέθηκε για την ΜΜΡ-9, δηλώνοντας ότι ίσως το ένζυμο αυτό να εμπλέκεται σε άλλες διεργασίες που επιτελούνται στο ΠΜ. / Serglycin (SG) is a proteoglycan expressed by hematopoietic cells and is constitutively secreted by multiple myeloma (MM) cells. SG participates in the regulation of various inflammatory events. We found that SG secreted by human MM cell lines inhibits both the classical and lectin pathways of complement, without influencing alternative pathway activity. It was also shown that SG could not initiate any activation of the complement system. The inhibitory effect of SG is due to direct interactions with C1q and mannose binding lectin (MBL). C1q-binding is mediated through the glycosaminoglycan moieties of SG, whereas binding to MBL requires the presence of SG protein core. Interactions between SG and C1q as well as MBL are diminished in the presence of chondroitin sulfate type E. In addition, we localized the SGbinding site to the collagen-like stalk of C1q. Interactions between SG and C1q as well as MBL are ionic in character and only the interaction with MBL was found to be partially dependent on the presence of calcium. Although we found the serum levels of SG to be elevated in patients with MM compared to healthy controls, no statistical significant differences were observed for classical and alternative pathway activity in sera between MM patients and healthy donors. Despite that, it is proved that increase levels of SG show a tendency to correlate with decreased levels of classical pathway activity in serum of MM patients. Moreover, we found that SG expressed from myeloma plasma cells protects these cells from complement activation induced by treatment with anti-thymocyte immunoglobulins. It is also demonstrated that SG on the surface of MM cells inhibits complement deposition on the membrane of these cells. Cells which do not express SG on its’ surface are 2-3 times more sensitive to complement attack compared with cells expressing high levels of SG. This might protect myeloma cells during immunotherapy and promote survival of malignant plasma cells. Moreover, it is shown that SG from MM cells is capable to interact with collagen type I, the most abundant collagen in bone. Furthermore, SG is present on the surface of myeloma plasma cells and it is involved in the adhesion of MM cells to collagen type I in bone marrow microenvironment. In addition, it is shown that the interaction of myeloma plasma cells to collagen type I, mediated by cell surface SG, induces expression and secretion of both MMP-2 and MMP-9 from MM cells. Along the process, we have investigated levels of MMP-2 and MMP-9 in serum and marrow of patients with MM. Decreased levels of both MMP-2 and MMP- 9 in serum of MM patients have been observed compared to healthy donors. Instead, MMP-2 was found to be increased in bone marrow of MM patients compared to control samples, while no differences observed for MMP-9. Finally, marrow levels of MMP-2 seem to correlate with serum levels of the enzyme along with a marker of bone resorption, NTx. This might indicate the implication of MMP-2 in the pathogenesis of bone disease in MM. Even though, no correlation of MMP-9 with NTx was observed, proving that MMP-9 may play a significant role in different pathogenetic mechanism occurring in MM.

Introdução: A Organização Mundial da Saúde estimou para o ano de 2008 aproximadamente 1.38 milhões de casos novos de câncer de mama no mundo e 458 mil mortes. A maioria dos casos (56%) e das mortes (64%), ocorrendo em países economicamente desenvolvidos. Apesar dos avanços e do diagnostico precoce, um número significativo de mulheres com tumores da mama operáveis apresentando evolução desfavorável vêm à sucumbir devido ao surgimento de doença metastática. Uma melhor compreensão da heterogeneidade do tumor e das características microambientais subjacentes, bem como dos mecanismos e as consequências das suas interações é essencial para melhorar o direcionamento das terapias existentes e desenvolver novos agentes terapêuticos para o câncer. Objetivos: Descrever as características clínicas, anatomopatológicas e imuno-histoquímicas de um grupo de pacientes com câncer de mama operável, e estudar o impacto destas características no estadiamento da doença, sobrevivência livre de recorrência e sobrevivência global. Além disto, analisar as potenciais correlações existentes entre estas características. Métodos: Estudo de coorte retrospectiva de base hospitalar envolvendo 86 mulheres com câncer primário de mama, submetidas a tratamento entre julho de 1999 e dezembro de 2004, no Serviço de Mastologia do Hospital de Clinicas de Porto Alegre. Dados clinicopatológicos e imuno-histoquimicos (RE, RP, HER2, Ki67 e p53) foram coletados dos registros hospitalares. Expressão do VEGF, MMP-2, MMP-9, TIMP-1 e TIMP-2 foram analisadas através de imuno-histoquimica. Variáveis contínuas foram analisadas pelo coeficiente de correlação de Spearman, ou pelo teste não paramétrico U de Mann-Whitney e H de Kruskal-Wallis, quando comparadas com variáveis categóricas. Variáveis categóricas foram analisadas pelo teste 2 de Pearson. Estimativas da probabilidade de sobrevivência foram obtidas pelo estimador não paramétrico de Kaplan-Meier e pelo semiparamétrico modelo de regressão de Cox. Comparação entre as curvas de sobrevivência foi realizada pelo teste estatístico de log-rank. O IC foi calculado em 95% e valores p< 0,05 foram considerados estatisticamente significativos. Resultados: A sobrevivência livre de recorrência em 5 e 10 anos foi de 82,2% e 68%, e a global foi de 90,2% e 82,9%, respectivamente. Número de linfonodos positivos (p= 0,00; p= 0,03), diâmetro tumoral (p= 0,01; p= 0,01) e estádio (p= 0,00; p= 0,02) são fatores de risco isolado para recorrência e óbito, respectivamente. Superexpressão de HER2 é um fator de risco isolado para recorrência (p= 0,04). Existe uma correlação positiva significativa entre: VEGF e MMP-9 (rs: 0,246; p= 0,023); TIMP-2 e MMP-2 (rs: 0,358; p= 0,001). Também foram encontradas associações significativas entre as variáveis: a) VEGF e receptor de progesterona positivo (p= 0,045); b) TIMP-2 e idade ≥ 50 anos (p= 0,002), e diâmetro ≤ 2,0 cm (p= 0,016); c) TIMP-1 e menarca ≤ 12 anos (p= 0,038); d) Maior diâmetro e alto grau histológico (2: 19,3; p= 0,004), invasão vascular (2: 12,6; p= 0,006), status do linfonodo axilar (2: 8,6; p= 0,035), número de linfonodos metastáticos (2: 7,2; p= 0,028), e recidiva a distancia (2: 4,0; p= 0,046); e) Invasão vascular e status do linfonodo axilar, e número de linfonodos metastáticos, ambos com 2: 24,7; p= 0,000. Conclusões: O número de linfonodos positivos, diâmetro tumoral, e estádio foram identificados como fatores de risco isolado para a ocorrência de recidiva e óbito. A superexpressão de HER2 é fator de risco isolado para a ocorrência de recidiva da doença. Novas pesquisas devem ser realizadas, com padronização de procedimento e um maior número de casos para melhor caracterização da doença. / Background: The World Health Organization estimated approximately 1.38 million new breast cancer cases and 458,000 deaths worldwide for 2008. Most of these (56% of new cases and 64% of deaths) occur in economically developed countries. In Brazil, approximately 52,000 new cases are predicted for 2013. Better understand the heterogeneity of the tumor and microenvironmental characteristics around you, as well as the mechanisms and consequences of their interactions is essential to improve the targeting of existing therapies and develop new therapeutic agents for cancer. Objectives: The aim of this study was to describe the clinical, anatomopathological and immunohistochemical characteristics of a group of patients with operable breast cancer, and investigate the impact of these characteristics on disease staging, disease-free survival and overall survival. In addition, the potential correlations between these characteristics were analyzed. Methods: This is a hospital-based retrospective cohort study of 86 women with primary breast cancer, subjected to surgical and adjuvant treatment between July 1999 and December 2004. Clinicopathological and immunohistochemical (ER, PR, HER2, Ki67 e p53) data were collected from hospital records. The expression of VEGF, MMP-2, MMP-9, TIMP-1 and TIMP-2 was analyzed using the immunohistochemical technique. Continuous variables were assessed with Spearman’s rank correlation coefficient, or the non-parametric Mann-Whitney U or Kruskal-Wallis H tests. Pearson’s 2 test was employed to asses categorical variables. The possibility of survival was estimated using the non-parametric Kaplan-Meier estimator and the semiparametric Cox regression model. Survival curves were compared using the statistical log-rank test. CI was calculated at 95% and p values <0.05 were considered statistically significant. Results: Disease-free survival at 5 and 10 years was 82.2% and 68%, and overall survival 90.2% and 82.9%, respectively. Number of positive lymph nodes (p= 0.00; p= 0.03), tumor diameter (p= 0.01; p= 0.01) and stage (p= 0.00; p= 0.02) were isolated risk factors for relapse and death, respectively. HER2 overexpression was an isolated risk factor for relapse (p= 0.04). There was a significant positive correlation between: VEGF and MMP-9 (rs: 0.246; p= 0.023) and TIMP-2 and MMP-2 (rs: 0.358; p= 0.001). Significant associations were also recorded between the following variables: a) VEGF and progesterone receptor-positive status (p= 0.045); TIMP-2 and age ≥ 50 years (p= 0.002) and diameter ≤ 2.0 cm (p= 0.016); c) TIMP-1 and menarche ≤ 12 years (p= 0.038); d) greater diameter and high histologic grade (2: 19.3; p= 0.004), vascular invasion (2: 12.6; p= 0.006), axillary lymph node status (2: 8.6; p= 0.035), number of metastatic lymph nodes (2: 7.2; p= 0.028) and distant relapse (2: 4.0; p= 0.046); e) vascular invasion and axillary lymph node status and number of metastatic lymph nodes, both with 2: 24.7 and p= 0.000. Conclusion: Number of positive lymph nodes, tumor diameter, and stage were identified as isolated risk factors for relapse and death. HER2 overexpression is an isolated risk factor for the occurrence of relapse. Further studies are needed, with standardization of the procedure and a larger number of cases, for better characterization of the disease.

Submitted by THALITA LEONE ALVES ROCHA null (thalarocha@hotmail.com) on 2016-04-18T15:51:39Z No. of bitstreams: 1 dissertação thalita final.pdf: 794073 bytes, checksum: 8f42b335b8638b50194b64ede638c446 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-04-19T16:16:26Z (GMT) No. of bitstreams: 1 rocha_tla_me_bot.pdf: 794073 bytes, checksum: 8f42b335b8638b50194b64ede638c446 (MD5) / Made available in DSpace on 2016-04-19T16:16:26Z (GMT). No. of bitstreams: 1 rocha_tla_me_bot.pdf: 794073 bytes, checksum: 8f42b335b8638b50194b64ede638c446 (MD5) Previous issue date: 2016-02-29 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / As metaloproteinases de matriz extracelular (MMPs) podem limitar a vasodilatação pulmonar e os efeitos inotrópicos positivos promovidos pela adrenomedulina durante a hipertensão pulmonar. O presente estudo teve por objetivo avaliar os efeitos da administração combinada da doxiciclina (inibidor não seletivo das MMPs) e da adrenomedulina sobre as alterações hemodinâmicas observadas durante a embolia pulmonar aguda em ovinos. Alterações hemodinâmicas e respiratórias foram mensuradas em ovinos anestesiados, pré-tratados com doxiciclina (10 mg/kg por via intravenosa), submetidos à EPA induzida pela injeção intravenosa (IV) de microesferas de silicone (500 mg) e posteriormente tratados com solução salina (grupo Dox+PE) ou adrenomedulina (50 ng/kg/min) (grupo Dox+PE+Adm). Os resultados deste estudo foram comparados com grupos históricos recentemente publicados por nosso grupo de pesquisa, realizados sob as mesmas condições experimentais, onde foram utilizados ovinos anestesiados não submetidos a qualquer intervenção (grupo Sham) ou submetidos à EPA e tratados com solução salina (grupo PE) ou com adrenomedulina (50 ng/kg/min) (grupo PE+Adm). Doxiciclina não produziu efeitos adicionais sobre as diminuições significativas no índice de resistência vascular pulmonar e aumento no índice cardíaco (ambos em 25%) observadas com o uso da adrenomedulina (grupo PE+Adm). A administração da adrenomedulina (grupo PE+Adm e Dox+PE+Adm) diminuiu significativamente a pressão arterial média e o índice de resistência vascular sistêmica, levando a uma hipotensão sistêmica moderada. Reduções significativas na pressão parcial de oxigênio arterial foram observadas após a doxiciclina e a EPA, que não foram afetadas pela administração da adrenomedulina. Estes resultados demonstram que a administração combinada da doxiciclina e adrenomedulina não traz benefícios hemodinâmicos adicionais quando comparada ao uso isolado da adrenomedulina, sugerindo que está combinação não se apresenta vantajosa durante a EPA induzida por microesferas. / Matrix metalloproteinases (MMPs) may limit severely the pulmonary vasodilatory and inotropic effects of adrenomedullin during pulmonary hypertension. Hemodynamic and respiratory changes were measured in anesthetized bovine pre-treated with doxycycline (10 mg/kg intravenously), subjected to APE induced by intravenous injection of silicone microspheres (500 mg) and subsequently treated with physiological saline (Dox+PE group) or adrenomedullin (50 ng / kg / min) (Dox+PE+ Adm group). The results were compared with historical group recently published by our research group, carried out under the same experimental conditions, where anesthetized sheep were used not subjected to any intervention (Sham group) or subjected to APE, and treated with physiological saline (PE group) or with adrenomedullin (50 ng / kg / min) (PE+Adm Group). Doxycycline produced no effect on significant temporal decreases in pulmonary vascular resistance index and increases in cardiac index (both by 25%) observed with adrenomedullin. The administration of adrenomedullin significantly decreased mean arterial pressure and systemic vascular resistance index, leading to a moderate systemic hypotension. Significant decreases in arterial oxygen partial pressure were observed after doxycycline or APE, but these changes were not affected by adrenomedullin. These results demonstrate that the combined administration of doxycycline and adrenomedullin does not provide additional hemodynamic benefits when compared to isolated use of adrenomedullin, suggesting that this combination does not appear advantageous for the APE-induced microspheres. / FAPESP: 2012/12.291-7

Orientador: Josimeri Hebling / Resumo: Objetivo: Avaliar o desempenho clínico de restaurações de resina composta em lesões cervicais não cariosas (LCNC) realizadas com ou sem a aplicação de um agente de ligação cruzada (carbodiimida, EDC) durante o período de 12 meses. Métodos: 21 pacientes com no mínimo duas LCNC participaram deste estudo. Um total de 142 LCNC foram distribuídas randomicamente em dois grupos (n=71 cada): controle e experimental. Todas as LCNC foram restauradas pelo mesmo operador e com os mesmos materiais (sistema adesivo Adper Single Bond 2 e resina composta Z350), sendo que, no grupo experimental a dentina recebeu a aplicação de EDC 0,5 mol/L por 60 segundos após o condicionamento ácido e no grupo controle apenas PBS. As restaurações foram avaliadas imediatamente após sua confecção (baseline) e após uma semana, 6 e 12 meses pelo critério USPHS/Ryge por apenas um avaliador calibrado. Os escores obtidos a partir da avaliação clínica, considerando-se os fatores de confundimento, foram analisados pelos testes de Woolf e Cochran-Mantel-Haenzel (p<0,05). Resultados: As taxas de falhas acumuladas na retenção das restaurações nos períodos 7 dias, 6 e 12 meses, foram de 0%; 3,0% e 4,5%, respectivamente, para o grupo controle e de 0%; 3,0% e 7,4% para o grupo experimental (EDC). Não houve diferença estatisticamente significante entre os dois grupos para todos os critérios e períodos avaliados. A taxa de retorno dos pacientes nos períodos de 7 dias, 6 e 12 meses foi de 100%, 90,5% e 85,7%, respectivamente... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: o evaluate the clinical performance of composite restorations placed in noncarious cervical lesions (NCCL) performed with or without the application of a cross-linking agent, over a period of 12 months. Methods: 21 patients with at least two NCCL, participated in this study. A total of 142 NCCL were randomly allocated into two groups (n=71): control and experimental. All NCCL were restored by the same operator and with the same materials (Single Bond 2 and Z350). In the experimental group the etched dentin was exposed to 0.5 mol/L EDC for 60 seconds while in the control group the etched dentin was exposed to Phosphate buffer solution (PBS). The restorations were evaluated at baseline, 7 days, 6 and 12 months using the USPHS/Ryge criteria, by one blinded calibrated examiner. The scores from the clinical evaluation were submitted to Woolf and CochranMantel-Haenzel tests, considering the confounding factors (p<0.05). Results: The rates of accumulated failures in the retention of restorations in the periods 7 days, 6 and 12 months were 0%; 3,0% and 4.5%, respectively, for the control group, and 0%; 3,0% and 7.4% for the experimental group. There was no statistically significant difference between the two groups for all criteria and periods. The return rate of patients in the 7-day, 6- and 12-month periods was 100%, 90.5% and 85.7%, respectively. Conclusion: The treatment of the etched dentin with with EDC prior to the application of the dentin bonding agent did not affect the performance of composite resin restorations in NCCL...(Complete abstract electronic access below) / Doutor

Made available in DSpace on 2014-06-11T19:23:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-04-07Bitstream added on 2014-06-13T19:09:51Z : No. of bitstreams: 1 laskoski_lm_me_jabo.pdf: 647030 bytes, checksum: 950b1f5d5358b98ef4e5d3a67333637b (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A laminite está associada a ativação inflamatória sistêmica, a qual ocorre em diversas afecções, principalmente as gastrointestinais. Sua patogenia este relacionada a ativação de metaloproteinases e degradação do colágeno e/ou hemidesmossomos. 0 objetivo deste estudo foi analisar alterações morfológicas, apoptose, metaloproteinases (MMP) 2 e 9 e infiltração leucocitária, pela pesquisa da lipocalina liberada por neutrófilos (NGAL), no tecido laminar de equinos submetidos a obstrução intestinal experimental. Os animais foram distribuídos nos grupos: controle (Gc), instrumentado (Gi), com procedimento de enterotomia sem obstrução; obstruído tratado com hidrocortisona (Gt) e obstruído não tratado (Gnt). 0 tecido laminar foi analisado pelas técnicas de microscopia em luz, imunohistoquímica e zimografia. Os animals dos grupos experimentais (Gi, Gt e Gnt) demonstraram alterações morfológicas, MMPs e substância formada pela M M P-9 e NGAL superiores aos resultados do Cc. A presença de apoptose não foi evidenciada pelo procedimento cirúrgico. 0 Gc não foi diferente quanto as alterações morfológicas quando comparado ao Gi, e também não diferiu do Gt em relação e MMP-9, principalmente associada a neutrófilos. Com estes resultados, conclui-se que: o modelo experimental pode causar alterações morfológicas associadas a laminite; a apoptose não participa da fase inicial de alterações laminares no córium laminar; a hidrocortisona pode ser utilizada para coibir os efeitos deletérios da ativação inflamatória associada a diversas doenças, sem comprometer o córium laminar. / The laminitis is associated to systemic inflammatory activation that occur in several diseases, mainly gastrointestinal. The pathogeny of the laminitis is related to matrix metalloproteinases (MMPs) activation with collagen or hemidesmossomos degradation. The aim of this study is evaluated of the morphologic alterations, apoptosis, MMPs 2 and 9 and leukocyte infiltration, for research of MMP-9 related to neutrophil gelatinase associated to lipocalin (NGAL) in laminar corium of horses subjected to intestinal obstruction experimental. The animals were divided in groups: control (Cc), instrumented (Gi), with enterotomy without obstruction; obstruction treated by hydrocortisone (Gt) and without treatment (Gnt). The laminar tissue was analyzed by optical microscopy, immunohistochemistry and zymography. The animals showed morphologic alterations, MM Ps expression and MMP-9 associated to NGAL greater the Gc. The presence of apoptosis was not evidend by cirurgical procedure. However, the Gc was not different to Gi about morphologic alterations, and too Gt, about MMP-9, mainly the form conjugated to NGAL. The results indicate that the experimental model can cause morphologic alterations associated with laminitis. The apoptosis is not prerequisite for laminar lesion. The hydrocortisone can be administrated to reduce the deleterious effects of inflammatory activation in several diseases, without to affect the laminar corium.

Orientador: Carlos Alan Candido Dias Junior / Resumo: As metaloproteinases de matriz extracelular (MMPs) podem limitar a vasodilatação pulmonar e os efeitos inotrópicos positivos promovidos pela adrenomedulina durante a hipertensão pulmonar. O presente estudo teve por objetivo avaliar os efeitos da administração combinada da doxiciclina (inibidor não seletivo das MMPs) e da adrenomedulina sobre as alterações hemodinâmicas observadas durante a embolia pulmonar aguda em ovinos. Alterações hemodinâmicas e respiratórias foram mensuradas em ovinos anestesiados, pré-tratados com doxiciclina (10 mg/kg por via intravenosa), submetidos à EPA induzida pela injeção intravenosa (IV) de microesferas de silicone (500 mg) e posteriormente tratados com solução salina (grupo Dox+PE) ou adrenomedulina (50 ng/kg/min) (grupo Dox+PE+Adm). Os resultados deste estudo foram comparados com grupos históricos recentemente publicados por nosso grupo de pesquisa, realizados sob as mesmas condições experimentais, onde foram utilizados ovinos anestesiados não submetidos a qualquer intervenção (grupo Sham) ou submetidos à EPA e tratados com solução salina (grupo PE) ou com adrenomedulina (50 ng/kg/min) (grupo PE+Adm). Doxiciclina não produziu efeitos adicionais sobre as diminuições significativas no índice de resistência vascular pulmonar e aumento no índice cardíaco (ambos em 25%) observadas com o uso da adrenomedulina (grupo PE+Adm). A administração da adrenomedulina (grupo PE+Adm e Dox+PE+Adm) diminuiu significativamente a pressão arterial média e o índice ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Matrix metalloproteinases (MMPs) may limit severely the pulmonary vasodilatory and inotropic effects of adrenomedullin during pulmonary hypertension. Hemodynamic and respiratory changes were measured in anesthetized bovine pre-treated with doxycycline (10 mg/kg intravenously), subjected to APE induced by intravenous injection of silicone microspheres (500 mg) and subsequently treated with physiological saline (Dox+PE group) or adrenomedullin (50 ng / kg / min) (Dox+PE+ Adm group). The results were compared with historical group recently published by our research group, carried out under the same experimental conditions, where anesthetized sheep were used not subjected to any intervention (Sham group) or subjected to APE, and treated with physiological saline (PE group) or with adrenomedullin (50 ng / kg / min) (PE+Adm Group). Doxycycline produced no effect on significant temporal decreases in pulmonary vascular resistance index and increases in cardiac index (both by 25%) observed with adrenomedullin. The administration of adrenomedullin significantly decreased mean arterial pressure and systemic vascular resistance index, leading to a moderate systemic hypotension. Significant decreases in arterial oxygen partial pressure were observed after doxycycline or APE, but these changes were not affected by adrenomedullin. These results demonstrate that the combined administration of doxycycline and adrenomedullin does not provide additional hemodynamic benefits when compared to iso... (Complete abstract click electronic access below) / Mestre