Submitted by PPG Engenharia e Tecnologia de Materiais (engenharia.pg.materiais@pucrs.br) on 2018-08-08T14:30:34Z No. of bitstreams: 1 Disserta??o Luisa Alban.pdf: 1572475 bytes, checksum: 51e47c0f80b15d05eda24af2e3b8fde5 (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-08-09T11:13:00Z (GMT) No. of bitstreams: 1 Disserta??o Luisa Alban.pdf: 1572475 bytes, checksum: 51e47c0f80b15d05eda24af2e3b8fde5 (MD5) / Made available in DSpace on 2018-08-09T11:23:16Z (GMT). No. of bitstreams: 1 Disserta??o Luisa Alban.pdf: 1572475 bytes, checksum: 51e47c0f80b15d05eda24af2e3b8fde5 (MD5) Previous issue date: 2018-08-01 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The interest in nanostructures such as titanate nanotubes (TNTs) has grown notably in recent years due to their biocompatibility and economic viability, which makes them promising for application in the biomedical field. Quercetin (Qc) has been reported to have great potential as a chemopreventive agent widely used in the study of the treatment of diseases such as bladder cancer. Therefore, this work aimed to study the incorporation of quercetin in sodium TNTs (NaTNT) and zinc (ZnTNT), as well as characterize the nanostructures formed. In addition, it was intended to conduct Qc release tests and biological and antitumor activities in T24 lineage cells. The nanostructures of TNTs were synthesized and characterized by FTIR, MEVFEG, EDS, MET DRX and TGA techniques. The results showed that the nanostructures have a tubular structure and the exchange of Na+ ions by Zn2+, as well as the incorporation of quercetin in the structure do not alter this morphology. In addition, the interaction established between Zn and Qc increases the thermal stability of nanostructures. The release test showed that the maximum delivery of Qc occurs after 24h and the presence of Zn controls the release of the flavonoid. Biological assays have shown that the NaTNTQc and ZnTNTQc nanostructures decrease the cellular viability of T24 after 48h in high concentrations. Furthermore, NaTNT, NaTNTQc and ZnTNT reduce the number of T24 lineage cells when combined with irradiation after 48h showing that the combination of nanostructures and ionizing energy is an attractive object of study in the treatment of bladder cancer. / O interesse por nanoestruturas como nanotubos de titanatos (TNTs) tem crescido notavelmente nos ?ltimos anos devido a sua biocompatibilidade e viabilidade econ?mica, o que os tornam promissores para a aplica??o na ?rea biom?dica. A quercetina (Qc) tem sido relatada como tendo grande potencial como agente quimiopreventivo amplamente utilizada no estudo do tratamento de doen?as como c?ncer de bexiga. Neste sentido, este trabalho pretendeu estudar a incorpora??o de quercetina em TNTs de s?dio (NaTNT) e de zinco (ZnTNT), bem como, realizar a caracteriza??o das nanoestruturas formadas. Al?m disso, pretendeu-se conduzir testes de libera??o de Qc e atividades biol?gicas e antitumorais em c?lulas da linhagem T24. As nanoestruturas de TNTs foram sintetizadas e caracterizadas por t?cnicas de FTIR, MEV-FEG, EDS, MET DRX e TGA. Os resultados mostraram que as nanoestruturas apresentam estrutura tubular, e a troca de ?ons Na+ por Zn2+, bem como a incorpora??o de quercetina na estrutura n?o alteram esta morfologia. Al?m disso, a intera??o estabelecida entre o Zn e Qc aumenta a estabilidade t?rmica das nanoestruturas. O ensaio de libera??o mostrou que a entrega m?xima de Qc ocorre ap?s 24h e apresen?a de Zn controla a libera??o do flavonoide para o meio. Os ensaios biol?gicos mostraram que as nanoestruturas NaTNTQc e ZnTNTQc diminuem a viabilidade celular de T24 ap?s 48h em altas concentra??es. Ainda, NaTNT, NaTNTQc e ZnTNT reduzem o n?mero de c?lulas da linhagem T24 quando combinadas com irradia??o ap?s 48h mostrando que a combina??o entre as nanoestruturas e energia ionizante se apresenta como um objeto de estudo atrativo no tratamento de c?ncer de bexiga.

<p>The two purposes of this thesis were to evaluate new diagnostic techniques of lymphnode staging in invasive bladder cancer and to evaluate the results of neoadjuvant chemotherapy in invasive bladder cancer.</p><p>Sentinel node detection was performed in 13 patients in preparation for radical cystectomy. The method showed to be feasible, and the results displayed the occurrence of metastatic nodes outside the traditional area of diagnostic dissection in a majority of patients. Four patients were metastasized, each one with one metastatic node detected with the help of the sentinel node procedure.</p><p>Four randomly selected sentinel nodes from four different unmetastasized patients were compared to the four metastatic sentinel nodes from the first series. After microdissection, p53 genomic structure, immunohistochemical expression and MVD (microvessel density) were assessed in the primary tumors and corresponding sentinel nodes. The results suggested that invasive bladder cancer mainly involved monoclonal proliferation with predominantly homogenous biomarker profile, but there were also signs of clonal evolution.</p><p>The Nordic Cystectomy Trial 2 (NCT2), is a randomized prospective trial investigating the possible benefit of neoadjuvant chemotherapy versus cystectomy only, in 311 eligible patients with urinary bladder cancer T2-T4aNXM0.Evaluation of overall survival did not show any statistically significant benefit in the experimental arm. This probably due to lack of statistical power.</p><p>To increase the statistical power we performed a combined analysis of randomized patients from both the Nordic Cystectomy Trial 1 (NCT1) and NCT2, n = 620. Eligible patients from NCT1 had T1G3, T2-T4a NXM0 urinary bladder cancer. Standard meta-analysis methods were used. The only end-point analysed was overall survival. Neoadjuvant platinum based combination therapy was associated with a 20 % reduction in the relative hazard in probability of death.</p>

<p>Wegener´s granulomatosis (WG) is an unusual, serious, systemic vasculitis with specific clinical findings. The studies in this thesis aim at broadening our understanding of the aetiology and outcome of WG.</p><p>Patients with WG were identified in the In-patient Register 1975-2001. During this time the incidence increased three-fold, and neither ANCA-related increased awareness, nor diagnostic drift, seem to fully explain this trend, but it is still unclear if a true rise in incidence exists. </p><p>Anti- neutrophil cytoplasmic antibodies (ANCA) have been presented as highly specific for vasculitis. In a series of consecutive cANCA/PR3-ANCA positive patients, we investigated the positive predictive value for ANCA, and the outcome of patients with a positive cANCA/PR3-ANCA but not vasculitis. These patients have a low future risk of developing vasculitis, possibly indicating that ANCA, in this setting, reflects neutrophil activating properties not specific to vasculitis.</p><p>By linkage of the WG-cohort, and randomly selected population controls, to the Multi-generation register, we identified all first-degree relatives and spouses of patients and controls, totally encompassing some 2,000 patients and 70,000 relatives. Familial aggregation of WG was the exception, with absolute risks of < 1 per 1000.However, relative risks in first-grade relatives amounted to 1.56 (95% CI 0.35-6.90) such that a moderate familial aggregation cannot be excluded.</p><p>In the WG-cohort, cancer occurrence and risk was compared to that of the general population. Patients with WG have an overall doubled risk of cancer, with particularly increased risks of bladder-cancer, haematopoietic cancers including lymphomas and squamous skin-cancer. In a case-control study nested within the WG-cohort, treatment with cyclophosphamide was compared among bladder-cancer patients and matched cancer-free controls. Absolute risk of bladder cancer as high as 10% some years after diagnosis were found, and this risk can partly be attributed to cyclophosphamide-treatment, with a dose-response relationship.</p>

The two purposes of this thesis were to evaluate new diagnostic techniques of lymphnode staging in invasive bladder cancer and to evaluate the results of neoadjuvant chemotherapy in invasive bladder cancer. Sentinel node detection was performed in 13 patients in preparation for radical cystectomy. The method showed to be feasible, and the results displayed the occurrence of metastatic nodes outside the traditional area of diagnostic dissection in a majority of patients. Four patients were metastasized, each one with one metastatic node detected with the help of the sentinel node procedure. Four randomly selected sentinel nodes from four different unmetastasized patients were compared to the four metastatic sentinel nodes from the first series. After microdissection, p53 genomic structure, immunohistochemical expression and MVD (microvessel density) were assessed in the primary tumors and corresponding sentinel nodes. The results suggested that invasive bladder cancer mainly involved monoclonal proliferation with predominantly homogenous biomarker profile, but there were also signs of clonal evolution. The Nordic Cystectomy Trial 2 (NCT2), is a randomized prospective trial investigating the possible benefit of neoadjuvant chemotherapy versus cystectomy only, in 311 eligible patients with urinary bladder cancer T2-T4aNXM0.Evaluation of overall survival did not show any statistically significant benefit in the experimental arm. This probably due to lack of statistical power. To increase the statistical power we performed a combined analysis of randomized patients from both the Nordic Cystectomy Trial 1 (NCT1) and NCT2, n = 620. Eligible patients from NCT1 had T1G3, T2-T4a NXM0 urinary bladder cancer. Standard meta-analysis methods were used. The only end-point analysed was overall survival. Neoadjuvant platinum based combination therapy was associated with a 20 % reduction in the relative hazard in probability of death.

This thesis is composed of three studies pertaining to the quality of care for radical cystectomy in Ontario between 1992 and 2004. In the first paper, the associations between provider volume and both operative and overall mortality were assessed. In the second paper, potential factors that could explain the association between volume and outcome were explored. In the final paper, the impact of waiting for cystectomy on survival outcomes was evaluated. Methods: A total of 3296 patients undergoing cystectomy for bladder cancer in Ontario between 1992 and 2004 were identified using the Canadian Institute for Health Information Discharge Abstract Database and the Ontario Cancer Registry. The effects of hospital and surgeon volume on operative mortality and overall survival were assessed using random effects logistic regression and marginal Cox Proportional Hazards modeling, respectively. To elucidate the factors underlying the volume-outcome association, the ability of a number of structure and process of care variables to attenuate the impact of volume was assessed. The effect of waiting for care, from transurethral resection to cystectomy, on overall survival was also assessed using marginal Cox models. Results: Neither hospital nor surgeon volume was significantly associated with operative mortality; however, both were associated with overall mortality. Of the measured structure/process measures, hospital factors caused the greatest attenuation of the volume hazard ratios, albeit to a limited degree. The wait time between the decision for surgery and cystectomy was also significantly associated with overall survival. The impact of delayed care was greatest for patients with lower stage disease. The data suggested a maximum wait time of 40 days for cystectomy. Conclusions: In this thesis, gaps in the quality of care for radical cystectomy in Ontario were identified. Patients treated by low volume hospitals and surgeons or those with long wait times all experienced worse outcomes. Since the underlying measures responsible for provider volume remain elusive, additional work is required to understand what these factors are. Initiatives to decrease wait times, however, are under way in Ontario. Whether these interventions decrease wait times and benefit patients remains to be seen.

Bakgrund: Urinblåsecancer finns i olika former och vid muskelinvasiv cancer är radikal operation den vanligaste behandlingen. Förutsatt att cancern inte spridit sig, då görs en cystektomi. Många människor har en naturlig förmåga att hantera kriser och skapar ny balans i tillvaron efter en påfrestande händelse i livet. Sjuksköterskan kan ge råd och stöd i situationen och bör då ha kunskap om förändringar i personens fysiska och psykiska tillstånd. Syfte: Syftet var att belysa hur personer som genomgått cystektomi på grund av urinblåsecancer upplevde att det dagliga livet påverkades. Metod: En litteraturstudie har gjorts där 15 vetenskapliga artiklar har granskats. Resultat: Personerna drabbades av olika problem efter cystektomin. Urinproblem, buk- och avföringsproblem och livsförändring var tydliga, men även problem med en förändrad kroppsuppfattning och problem med det sociala stödet kom fram. Slutsats: Det är olika faktorer som påverkar det dagliga livet efter en cystektomi. Kunskap i hur personerna kan reagera är viktig för sjuksköterskan, för att kunna bemöta problemen och ge råd och stöd om olika copingstrategier för att hantera situationen. Då kan individuell omvårdnad ges, utifrån varje persons behov. / Background: Bladder cancer exists in different forms and in muscle invasive cancer, radical surgery is the standard treatment. Assuming that the cancer has not spread, then, a cystectomy is done. Many people have a natural ability to handle crises and generate a new personal balance. A nurse can offer advice and support in the situation and should have knowledge of changes in the person's physical and mental condition. Aim: The aim was to illustrate how people who have undergone cystectomy because of bladder cancer felt that their daily lives were affected. Methods: A literature review has been done where 15 scientific papers have been reviewed. Results: The subjects suffered from various problems after cystectomy. Urinary Problems, abdominal and stool problems, and change in life were common, but also problems with an altered body image and problems with social support occurred. Conclusion: There are different factors that affect the daily lives after a cystectomy. Knowledge of how people might react is important for nurses, in order to address the problems and provide advice and support on various coping strategies to deal with the situation. When that is the case, individual care can be given, based on each person's needs.

This thesis is composed of three studies pertaining to the quality of care for radical cystectomy in Ontario between 1992 and 2004. In the first paper, the associations between provider volume and both operative and overall mortality were assessed. In the second paper, potential factors that could explain the association between volume and outcome were explored. In the final paper, the impact of waiting for cystectomy on survival outcomes was evaluated. Methods: A total of 3296 patients undergoing cystectomy for bladder cancer in Ontario between 1992 and 2004 were identified using the Canadian Institute for Health Information Discharge Abstract Database and the Ontario Cancer Registry. The effects of hospital and surgeon volume on operative mortality and overall survival were assessed using random effects logistic regression and marginal Cox Proportional Hazards modeling, respectively. To elucidate the factors underlying the volume-outcome association, the ability of a number of structure and process of care variables to attenuate the impact of volume was assessed. The effect of waiting for care, from transurethral resection to cystectomy, on overall survival was also assessed using marginal Cox models. Results: Neither hospital nor surgeon volume was significantly associated with operative mortality; however, both were associated with overall mortality. Of the measured structure/process measures, hospital factors caused the greatest attenuation of the volume hazard ratios, albeit to a limited degree. The wait time between the decision for surgery and cystectomy was also significantly associated with overall survival. The impact of delayed care was greatest for patients with lower stage disease. The data suggested a maximum wait time of 40 days for cystectomy. Conclusions: In this thesis, gaps in the quality of care for radical cystectomy in Ontario were identified. Patients treated by low volume hospitals and surgeons or those with long wait times all experienced worse outcomes. Since the underlying measures responsible for provider volume remain elusive, additional work is required to understand what these factors are. Initiatives to decrease wait times, however, are under way in Ontario. Whether these interventions decrease wait times and benefit patients remains to be seen.

El cáncer de vejiga es una enfermedad heterogénea. Los tumores se distribuyen en dos vías con cierto nivel de solapamiento: la vía papilar superficial caracterizada por alteraciones de FGFR3 y pérdida del cromosoma 9 y la vía no papilar invasiva caracterizada por alteraciones en las vías de p53 y pRb. Los tumores T1G3 representan el 10% de los tumores de vejiga diagnosticados y representan un desafío clínico debido a su alto riesgo de progresión y la falta de marcadores moleculares que predigan el pronóstico de los pacientes. El objetivo de la tesis ha sido caracterizar los tumores T1G3 y asociar los marcadores evaluados con el pronóstico de los pacientes. La caracterización de los tumores T1G3 ha identificado que la vía de p53 está alterada en un 85% de los casos, que los tumors muestran altos niveles de inestabilidad genómica y que tan sólo el marcador de inestabilidad FGA predice el pronóstico de los pacientes con tumores T1G3. Un nuevo marcador del cancer de vejiga se ha identificado: PIK3CA. / Bladder cancer is a heterogeneous disease distributed into two distinct but slightly overlapping pathways: a papillary superficial pathway characterized by alterations in FGFR3 and loss of chromosome 9 and a non-papillary invasive pathway characterized by alterations in the p53 and pRb pathways. T1G3 tumors are a subgroup of bladder cancers which represent 10% of diagnosed tumors and are a clinical challenge due to their high risk of progression and the lack of molecular markers to predict the prognosis of the patients. The aim of this thesis was to characterize T1G3 tumors and associate these markers with the outcome of the patients. The characterization of T1G3 tumors have shown that the p53 pathway is inactive in 85% of patients, that they have high levels of genomic instability and that only FGA predicts outcome among patients with T1G3 tumors. A novel marker for bladder cancer has been identified: PIK3CA.

Wegener´s granulomatosis (WG) is an unusual, serious, systemic vasculitis with specific clinical findings. The studies in this thesis aim at broadening our understanding of the aetiology and outcome of WG. Patients with WG were identified in the In-patient Register 1975-2001. During this time the incidence increased three-fold, and neither ANCA-related increased awareness, nor diagnostic drift, seem to fully explain this trend, but it is still unclear if a true rise in incidence exists. Anti- neutrophil cytoplasmic antibodies (ANCA) have been presented as highly specific for vasculitis. In a series of consecutive cANCA/PR3-ANCA positive patients, we investigated the positive predictive value for ANCA, and the outcome of patients with a positive cANCA/PR3-ANCA but not vasculitis. These patients have a low future risk of developing vasculitis, possibly indicating that ANCA, in this setting, reflects neutrophil activating properties not specific to vasculitis. By linkage of the WG-cohort, and randomly selected population controls, to the Multi-generation register, we identified all first-degree relatives and spouses of patients and controls, totally encompassing some 2,000 patients and 70,000 relatives. Familial aggregation of WG was the exception, with absolute risks of &lt; 1 per 1000.However, relative risks in first-grade relatives amounted to 1.56 (95% CI 0.35-6.90) such that a moderate familial aggregation cannot be excluded. In the WG-cohort, cancer occurrence and risk was compared to that of the general population. Patients with WG have an overall doubled risk of cancer, with particularly increased risks of bladder-cancer, haematopoietic cancers including lymphomas and squamous skin-cancer. In a case-control study nested within the WG-cohort, treatment with cyclophosphamide was compared among bladder-cancer patients and matched cancer-free controls. Absolute risk of bladder cancer as high as 10% some years after diagnosis were found, and this risk can partly be attributed to cyclophosphamide-treatment, with a dose-response relationship.

Methyl 2-cyano-3,11-dioxo-18b-olean-1,12-dien-30-oate (CDODA-Me) is a synthetic triterpenoid that inhibits growth of Panc1 and Panc28 pancreatic cancer cell lines and activates peroxisome proliferator-activated receptor B (PPARB)-dependent transactivation in these cells. CDODA-Me has also induced p21 and p27 protein expression and downregulated cyclin D1; however, these responses were receptor-independent. CDODA-Me induced apoptosis, which was accompanied by receptor-independent induction of the proapoptotic proteins early growth response-1 (Egr-1), nonsteroidal anti-inflammatory drug-activated gene-1 (NAG-1), and activating transcription factor-3 (ATF3). Induction of NAG-1 in Panc28 cells was p38-mitogen-activated protein kinase (MAPK) and phosphatidylinositol-3-kinase (PI3-K)-dependent, but Egr-1-independent, whereas induction in Panc1 cells was associated with activation of p38-MAPK, PI3-K and p42-MAPK and was only partially Egr-1-dependent. Specificity protein (Sp) transcription factors Sp1, Sp3 & Sp4 are overexpressed in multiple tumor types and negative prognostic factors for survival. Since Sp proteins regulate genes associated with survival (survivin), angiogenesis [vascular endothelial growth factor and its receptors] and growth [cyclin D1, epidermal growth factor receptor], research in this laboratory has focused on development of anticancer drugs that decrease Sp protein expression. Arsenic trioxide, curcumin, 2-cyano-3,12-dioxoleana-1,9-dien-28-oic acid (CDDO), CDDO-Me, and celastrol exhibit antiproliferative, antiangiogenic and proapoptotic activity in many cancer cells and tumors. Treatment of cancer cells derived from urologic and gastrointestinal tumors with arsenic trioxide decreased Sp1, Sp3 and Sp4 transcription factors and cotreatment with the proteosome inhibitor MG132 did not inhibit downregulation of Sp proteins in these cancer cells. Mechanistic studies suggested that compound-dependent downregulation of Sp and Sp-dependent genes was due to decreased mitochondrial membrane potential and induction of reactive oxygen species, and the role of peroxides in mediating these responses was confirmed using hydrogen peroxide, demonstrating that the mitochondriotoxic effects of these compounds are important for their anticancer activities. Moreover, repression of Sp and Sp-dependent genes by CDDO-Me and celastrol was due to downregulation of microRNA-27a and induction of ZBTB10, an Sp repressor, and these responses were also reversed by antioxidants. Thus, the anticancer activity of CDDO-Me and celastrol is due, in part, to activation of ROS which in turn targets the microRNA-27a:ZBTB10?Sp transcription factor axis to decrease growth inhibitory, pro-apoptotic and antiangiogenic genes and responses.