Made available in DSpace on 2015-04-14T13:35:24Z (GMT). No. of bitstreams: 1 436876.pdf: 1707187 bytes, checksum: a5856afc9a54d87e34f88e782140d450 (MD5) Previous issue date: 2012-01-20 / Gliomas are the most common and devastating tumors of the central nervous system (CNS). Among the gliomas group, the GBM is the most prevalent, aggressive and deadly malignant. Many pieces of evidence point out the relevance of natural compounds for cancer therapy and prevention, including chalcones. Chalcones are group of natural precursors of flavonoid and display a wide variety of biological and pharmacological proprieties that include anti-proliferative and anti-cancer activities. This study aimed at evaluating the in vitro anti-proliferative activity and cell viability inhibition of nine quinoxaline derived chalcones, structurally based on the selective PI3Ky inhibitor AS605240. These synthetic compounds were tested at different time-periods of incubation (24, 48 e 72 h) and concentrations (0,1; 0,1; 1; 5 e 10 μg/mL) in glioma cell lines from human and rat origin (U-138 MG and C6, respectively). The results showed by MTT assay and cell couting revealed that four chalcones (compounds N2, N9, N10 and N12), displayed higher efficacies and potencies, being able to inhibit either cell proliferation or viability, in a time- and concentration dependent manner. These four compounds which present methoxy groups at A-ring and their efficacy was greater than that seen for the positive control compound AS605240. Flow cytometry analysis demonstrated that incubation of C6 cells with chalcone N9 led to G1 phase arrest, likely indicating an interference with apoptosis. Furthermore, chalcone N9 was able to visibly inhibit AKT activation, allied to the stimulation of ERK 1/2 MAP-kinase. The chalcones tested herein, especially those displaying a methoxy substituent at A-ring, might well represent promising molecules for the treatment of gliomas. / Os gliomas s?o os tumores do SNC mais comuns e devastadores. Dentre os gliomas, o GBM ? o mais prevalente, agressivo, maligno e apresenta um mau progn?stico. A relev?ncia dos compostos naturais, incluindo as chalconas, no tratamento e na preven??o do c?ncer est? sendo muito evidenciada. As chalconas formam um grupo de compostos naturais derivados dos flavon?ides que apresentam diferentes propriedades biol?gicas e farmacol?gicas, incluindo as atividades anti-proliferativas e anti-tumorais. O objetivo deste estudo foi avaliar a a??o anti-proliferativa e a capacidade de inibi??o da viabilidade celular in vitro de nove chalconas derivadas da quinoxalina, baseadas estruturalmente no inibidor seletivo de PI3Ky, o AS605240. Estes compostos sint?ticos foram testados em diferentes tempos de incuba??o (24, 48 e 72 h) e concentra??es (0,1; 0,1; 1; 5 e 10 μg/mL) em linhagens de glioma humano e de rato (U-138 MG e C6, respectivamente). Os resultados observados nos experimentos de MTT e na contagem celular revelaram que quatros chalconas (compostos N2, N9, N10 e N12), apresentaram grande efic?cia e pot?ncia, sendo capazes de inibir a prolifera??o e a viabilidade celular, de maneira tempo e concentra??o dependente. Estes quatro compostos que possuem radicais met?xi no anel A de sua estrutura demonstraram uma efic?cia superior ?quela do composto AS605240, usado como controle positivo. Os resultados da citometria de fluxo demonstraram que a incuba??o das c?lulas C6 com a chalcona N9 levaram a um bloqueio celular na fase G1, possivelmente indicando interfer?ncia com a apoptose. Al?m disto, a chalcona N9 foi capaz de inibir visivelmente a ativa??o da AKT, aliada ? estimula??o de ERK 1/2 MAP-quinase. As chalconas estudadas neste projeto, especialmente as que apresentam o radical metoxi no anel A, representam promissoras mol?culas para o tratamento dos gliomas.

Made available in DSpace on 2015-04-14T13:35:27Z (GMT). No. of bitstreams: 1 437323.pdf: 4287591 bytes, checksum: a4736ecf5ed6c11a9d86f23c3c30d3b3 (MD5) Previous issue date: 2012-01-18 / Tuberculosis re-emerged in the mid 80s and currently about two million people die each year due to this disease. The resurgence of tuberculosis has become a threat to public health. The high susceptibility of HIV-infected patients and the proliferation of multi-drug resistant strains have created the need to develop new therapies. However, no new class of drugs against tuberculosis was developed in the last 45 years. Thus, it becomes imminent need to develop new antituberculosis agents. As purine metabolism could be implicated in mycobacterial latency the purine nucleoside phosphorylase (EC 2.4.2.1) has been proposed as target for development of antibacterial drugs. PNP catalyzes the reversible cleavage in the presence of inorganic phosphate of N-ribosidic bonds of the purine nucleosides and deoxynucleosides, except adenosine. This reaction generates the purine base and ribose (deoxyribose)-1-phosphate. PNP is specific for purine nucleosides in the β-configuration and cleaves the glycosidic bond with inversion of configuration to produce α-ribose-1-phosphate. In this work PNP was crystallized in association with inosine, hypoxanthine and acyclovir, substrate, product and inhibitor, respectively, and data were collected using synchrotron radiation. In addition, molecular dynamics simulations and isothermal titration calorimetry experiments were used to provide detailed information on the dynamic properties and to investigate the profile and thermodynamic affinities of MtPNP associated with these molecules. With obtained results we hope to contribute to the search of new selective inhibitors for MtPNP, since differences between the mycobacterial and human enzyme binding sites have been also identified, making structure-based drug design feasible. / A tuberculose ressurgiu na metade dos anos 80 e, atualmente aproximadamente dois milh?es de pessoas morrem por ano devido a esta doen?a. O ressurgimento da tuberculose tornou-se uma amea?a ? sa?de p?blica. A alta susceptibilidade dos pacientes infectados com HIV e a prolifera??o de cepas resistentes a m?ltiplas drogas t?m criado a necessidade de se desenvolver novas terapias. No entanto, nenhuma nova classe de drogas contra a tuberculose foi desenvolvida nos ?ltimos 45 anos. Deste modo, torna-se iminente a necessidade de se desenvolver novos agentes antituberculose. Como o metabolismo de purinas pode estar implicado na lat?ncia da micobact?ria, a purina nucleos?deo fosforilase de Mycobacterium tuberculosis (MtPNP) (EC 2.4.2.1) foi proposta como alvo para o desenvolvimento de drogas antibacterianas. A PNP catalisa a clivagem revers?vel, na presen?a de fosfato inorg?nico, de liga??es N-ribos?dicas de nucleos?deos e desoxinucleos?deos de purina, com exce??o da adenosina. Esta rea??o produz base p?rica livre e ribose (desoxiribose)-1-fosfato. A rea??o se processa com invers?o de configura??o, de β-nucleos?deos para α-ribose-1-fosfato. Neste trabalho a PNP foi cristalizada em associa??o com inosina, hipoxantina e aciclovir, substrato, produto e inibidor, respectivamente, e os dados foram coletados utilizando radia??o s?ncrotron. Al?m disso, a simula??o por din?mica molecular e experimentos de calorimetria por titula??o isot?rmica foram utilizadas para fornecer informa??es detalhadas acerca das propriedades din?micas, e investigar o perfil termodin?mico e afinidades da MtPNP associada com estas mol?culas. Com os resultados obtidos n?s esperamos contribuir para a busca de novos inibidores seletivos de MtPNP, j? que as diferen?as entre os s?tios de liga??o da enzima humana e de micobact?ria foram identificadas , tornando vi?veis projetos de desenho de drogas baseados na estrutura.

Made available in DSpace on 2015-04-14T13:35:32Z (GMT). No. of bitstreams: 1 438286.pdf: 4118090 bytes, checksum: 3f84072ed045d46706a18668827636e2 (MD5) Previous issue date: 2008-11-24 / Introduction: Previous studies showed a possible association between the T102C polymorphism of the serotonin 2A receptor (5-HT2A) and urinary incontinence in the elderly. Since serotonin is involved in the modulation of bladder sphincter control, such association is biologically plausible. Since the pharmacological treatment of urinary incontinence (UI) includes the use of serotonin reuptake inhibitors, complementary studies of this polymorphism are of scientific and clinical importance. Objectives: The aims of this study in older women living in a senior living community were: (1) to describe and compare the allelic and genotypic frequencies of the 102C polymorphism of the serotonin 2A receptor (5-HT2A) in incontinent and continent individuals; (2) to describe and compare the urodynamic parameters in incontinents carrying different genotypes/alleles; (3) to describe and compare the frequency of the urodynamic types of urinary incontinence (stress, urge and mixed) in incontinent individuals carrying different genotypes/alleles; and (4) to describe and compare the impact of UI on the quality of life in incontinent individuals carrying different genotypes/alleles. Methodology: A case-control study was conducted including 68 incontinent older women (submitted to urodynamic evaluation) and 162 continent older women (self-reporting). The criteria for exclusion were: use of diuretics, diabetes, non-Caucasian individuals, and severe mobility and neurological disturbances. The polymorphism was analyzed by the PCR-RFLP technique. The Ethics Committee approved the study, and all participants signed an informed consent form. Results: Incontinent older women showed a significantly greater frequency of the TT genotype (TT= 34.8%, CC= 19.3%, TC= 45.9%) compared to continent older women (TT=17%, CC=23.3%, TC=59.7%) and the general sample of the population (TT=21.5%, CC=16.6%, TC=61.9%) (p=0.001). The genetic frequencies of the three samples were in Hardy-Weinberg equilibrium. Urodynamic analysis showed a significant association between the TT genotype and greater detrusor pressure at maximal cystometric capacity, lower maximal cystometric capacity and reduced bladder compliance. The TT genotype also showed a higher prevalence of urge UI (44%) than did the other genotypes (p=0.01). Analyses of indicators of quality of life and UI demonstrated a greater negative impact in carriers of the C allele (CC+TC). Conclusion: Together, the results suggest that the T102C polymorphism of the 5-HT2A receptor gene has a physiological effect on the lower urinary tract of humans. This hypothesis needs to be confirmed by complementary studies. The effect of an imbalance in the quantity of this receptor, which occurs in the TT genotype, could be associated with a greater chance or urge UI in older women. / Introdu??o: estudos pr?vios mostraram poss?vel associa??o entre o polimorfismo T102C do receptor 2A da serotonina (5-HT2A) e incontin?ncia urin?ria em idosos. Como a serotonina est? envolvida na modula??o do controle vesical-esfincteriano, tal associa??o ? biologicamente plaus?vel. Uma vez que a terap?utica farmacol?gica da incontin?ncia urin?ria (IU) inclui o uso de inibidores da recapta??o da serotonina, estudos complementares deste polimorfismo s?o de relev?ncia cl?nico-cient?fica. Objetivos: em mulheres idosas que vivem na comunidade: (1) descrever e comparar as freq??ncias al?licas e genot?picas do polimorfismo 102C do receptor 2A da serotonina (5-HT2A) em idosas incontinentes e continentes; (2) descrever e comparar os par?metros urodin?micos nas idosas incontinentes portadoras de diferentes gen?tipos/alelos; (3) descrever e comparar a freq??ncia dos tipos de incontin?ncia urin?ria urodin?mica (de esfor?o, de incontin?ncia de urg?ncia e mista) nas idosas incontinentes portadoras de diferentes gen?tipos/alelos; (4) descrever e comparar o impacto da IU na qualidade de vida nas idosas incontinentes portadoras de diferentes gen?tipos/alelos. Metodologia: um estudo caso-controle foi conduzido, sendo 68 mulheres idosas incontinentes (submetidas ? avalia??o urodin?mica) e 162 mulheres continentes (auto-relato). Os crit?rios de exclus?o foram: uso de diur?ticos, diabetes, indiv?duos de outras etnias que n?o a caucasiana, dist?rbios de mobilidade e neurol?gicos graves. O polimorfismo foi analisado por t?cnica de biologia molecular PCR-RFLP. O Comit? de ?tica aprovou a pesquisa e todos os participantes assinaram Termo de Consentimento Livre e Esclarecido. Resultados: idosas incontinentes apresentaram freq??ncia do gen?tipo TT significativamente maior (TT= 34,8%, CC= 19,3%, TC= 45,9%) do que nas mulheres continentes (TT=17%, CC=23,3%, TC=59,7%) e na amostra geral da popula??o (TT=21,5%, CC=16,6%, TC=61,9%) (p=0,001). As freq??ncias gen?ticas das tr?s amostras estavam em equil?brio de Hardy-Weinberg. An?lise urodin?mica mostrou associa??o significativa entre o gen?tipo TT e maior press?o detrusora na capacidade cistom?trica m?xima, menor capacidade cistom?trica m?xima e complac?ncia vesical reduzida. O gen?tipo TT tamb?m apresentou maior preval?ncia de IU de urg?ncia (44%) do que os demais gen?tipos (p=0,01). O conjunto destes resultados sugeriu que o gen?tipo TT est? associado a altera??es de fatores fisiol?gicos, predisponentes a incontin?ncia urin?ria. Entretanto, an?lises de indicadores da qualidade de vida e a IU mostraram maior impacto negativo em portadoras do alelo C (CC+TC). Esta condi??o pode estar associada ? sugest?o de predisposi??o de portadoras do alelo C a dist?rbios emocionais. Conclus?o: o conjunto dos resultados sugere que o polimorfismo T102C do gene do receptor 5-HT2A tenha um poss?vel efeito fisiol?gico no trato urin?rio inferior (TUI) de seres humanos. Esta hip?tese precisa ser confirmada por estudos complementares. O efeito do desbalan?o na quantidade deste receptor, que ocorre no gen?tipo TT estaria associado ? maior chance de IU de urg?ncia na mulher idosa.

Made available in DSpace on 2015-04-14T13:35:33Z (GMT). No. of bitstreams: 1 438529.pdf: 18860537 bytes, checksum: 9c31f74e1e908515a22fbc3a7838b2b1 (MD5) Previous issue date: 2012-05-29 / Gastrointestinal sarcomatoid tumors (GIST) are rare malignancies, corresponding to 1 - 3% of alli GI cancers. Nevertheless, GISts are the most common GI mesenchimal malignancies. GISTs are heterogeneous in presentation, clinical course and response to therapy. Their prognosis can be roughly estimated at diagnosis by assessing tumor size, mitotic index and the topography of the primary lesion. Despite the proved utility of these prognostic factors, their limitations are well documented, and a more refined set of biological markers are much needed. The abnormal activation of the cell survival signaling pathway provided by PI3K/Akt/PTEN proteins is a well known feature in advanced epithelial and hematological malignancies, impacting the aggressiveness of the disease as well as the resistance to different traditional therapies or biological agents. Besides their multi-effect in cell survival mechanisms, the PI3K/Akt/PTEN pathway is a powerful activator of angiogenesis via VEGF protein family transcription. The different isoforms of VEGF will in turn activate the VEGF receptor (VEGFR) at the surface of endothelial cells. Therefore, VEGFR is a reliable marker of angiogenesis in human tumors. We sought to evaluate the expression of VEGFR and Akt proteins in a series of GIST patients at diagnosis, treated in a single institution, in order to correlated the levei of expression of these proteins to clinical outcomes, such as overall survival (OS) and disease free survival (DFS). The levei of expression of these proteins was also correlated to the same outcomes in a subgroup of patients with advanced disease who received more than three months of therapy with Imatinib Mesylate (GleeveC?). Methods - 48 GIST patients were evaluated. Immunohystochemistry for VEGFR and Akt was performed by tissue microarray. Results - Akt hyperexpression was statistically correlated with a shorter DFS in the group of patients with metastatic disease (ali of them receiving GleeveC?). This correlation was not observed in early cases (defined as curable disease - stages I to III). We have not observed correlation between Akt expression and OS in any clinical stage. Lower VEGFR expression was associated to a significant better OS in ali stages (including patients receiving GleeveC?). Conversely, DFS was not affected by the levei of expression of VEGFR. CONCLUSIONS - Low VEGFR expression was related to a better prognosis in any clinicalstage of GIST, reflected in larger OS. The fact that this effect was observed in GleeveC? treated patients suggests that the betler response to the drug is much more related to the biology of the tumor than to the treatment itself. Akt showed also a promising role as a prognostic marker for disease free survival in advanced disease, indicating that GISTs with hyperexpressed Akt pathway behave more aggressively or may be less sensitive to GleeveC?. / Introdu??o: GISTs s?o raros, correspondendo de 1 a 3% de todas as neoplasias do trato gastrointestinal, entretanto, s?o as neoplasias mesenquimais mais comuns desta localiza??o. Possuem uma apresenta??o, comportamento cl?nico e resposta ? terapia heterog?neos. O progn?stico pode ser grosseiramente estimado pelo tamanho tumoral ao diagn?stico, topografia e ?ndice mit?tico. Apesar da utilidade destes fatores de progn?stico de simples obten??o, a heterogeneidade da doen?a exige que marcadores mais espec?ficos sejam aplicados para uma avalia??o individual n?o apenas progn?stica ao diagn?stico, mas tamb?m preditiva ao tratamento com novas drogas dispon?veis. A ativa??o aberrante das prote?nas da via de sinaliza??o PI3K/Akt/PTEN ? um fen?meno relativamente comum em neoplasias epiteliais e hematol?gicas, com repercuss?o tanto na agressividade da neoplasia como na sua resist?ncia adquirida a tratamentos anti-neopl?sicos. Al?m da sinaliza??o para mecanismos de sobreviv?ncia, a via PI3K/Akt/PTEN est? envolvida na ativa??o de angiog?nese por meio da transcri??o de VEGF e secundariamente do seu receptor endotelial, VEGFR. O objetivo deste trabalho foi avaliar a express?o das prote?nas VEGFR, Akt e PTEN em pacientes com GIST ao diagn?stico, na tentativa de correlacionar a mesma com desfechos cl?nicos. Paralelamente acompanhamos o efeito do n?vel de express?o destas prote?nas em um subgrupo de pacientes com doen?a avan?ada que recebeu a medica??o Mesilato de Imatinib (GliveC?). M?todo: Avalia??o imunoistoqu?mica com m?todo TMA de 48 esp?cimes de GISTs utilizando os anticorpos monoclonais para Akt e VEGFR. A hiperexpress?o de Akt foi verificada em 33,3% e VEGFR em 75% das amostras. Na an?lise do grupo geral de pacientes, a hiper-express?o de Akt correlacionou-se com uma SLP (sobrevida livre de progress?o) estatisticamente encurtada (p>O.05) exclusivamente para casos com doen?a metast?tica. Esta correla??o n?o foi observada em casos com doen?a n?o inicial (est?gios I a III). No entanto, mesmo em doen?a avan?ada, a hiper-express?o de Akt n?o teve impacto na sobrevida global. Como o grupo de pacientes com doen?a avan?ada foi o grupo que recebeu GliveC?, a correla??o com SLP e hiper-express?o de Akt se manteve. A aus?ncia de express?o de VEGFR se associou com melhor sobrevida global (SG) em todos os est?gios (incluindo os pacientes que receberam GliveC?), mas n?o com sobrevida livre de doen?a (SLD). Este dado sugere que os pacientes com baixa express?o de VEGFR poderiam ter uma doen?a de biologia mais indolente, mesmo que a taxa de recorr?ncia seja id?ntica ao grupo com VEGFR hiper-expresso. Conclus?es: os resultados deste estudo sugerem que a baixa express?o de VEGFR ? um fator de progn?stico favor?vel em pacientes com doen?a em qualquer est?gio. A hiperexpress?o de Akt associa-se a uma menor SLP apenas em casos avan?ados, sugerindo que a ativa??o desta via poderia ser um mecanismo adquirido em etapas finais da doen?a que conferem resist?ncia ao GliveC?.

Made available in DSpace on 2015-04-14T13:35:57Z (GMT). No. of bitstreams: 1 458592.pdf: 13592524 bytes, checksum: 0f4861417438fcd80d410bc2aaf21c00 (MD5) Previous issue date: 2014-02-24 / Purpose: Increased expression of tissue factor (TF) was correlated with worse prognosis and survival in various malignant neoplasms. In urologic tumors, it was studied in prostate, bladder and kidney cancers. The present study evaluates the immunohistochemical expression of TF in human penile cancer, correlating it with prognostic factors and survival. Materials and Methods: A total of 34 patients were treated by partial or total penectomy for penile carcinoma at our institution from January 1998 to December 2012. The immunohistochemical expression of TF was determined by a quantitative methodology in formalin-fixed paraffin-embedded tissue samples and correlated with age, clinical stage, pathologic stage, lymph node metastases, tumor grade and survival. Results: Immunohistochemical expression of TF was observed in 26/34 (76.4%) specimens. The mean area of TF expression in the samples was 27.82 μm2 [95% CI 20.0 35.6, SD 22.4]. Mean age was significantly lower among patients with increased TF expression in comparison to those with lower TF expression [57.6 (SD 9,9) vs. 67.3 years (SD 16,4); p = 0.047). Mean postoperative follow-up was 28.3 months (from 1 to 139.6 months). There were eight deaths (23.5%), four in each group, all related to the penile cancer. In multivariate analyses, there was no statistically significant correlation between TF expression and the study variables. Conclusions: TF was expressed in most samples of penile cancers. There was no statistically significant correlation between TF expression, prognostic factors and survival. / Embasamento: O aumento da express?o do fator tecidual (FT) foi correlacionado com pior progn?stico e sobrevida em v?rias neoplasias malignas. Em tumores urol?gicos, foi estudado na pr?stata, bexiga e rim. No presente estudo avaliamos a express?o imunoistoqu?mica do FT em esp?cimes de c?ncer de p?nis, correlacionando com fatores progn?sticos e sobrevida. Materiais e M?todos: Um total de 34 pacientes tratados com penectomia parcial ou total por c?ncer de p?nis no Hospital S?o Lucas da PUCRS no per?odo entre janeiro de 1998 a dezembro de 2012 foram inclu?dos no estudo. A express?o do FT foi determinada com metodologia quantitativa em amostras de tecido em parafina e correlacionada com idade, estadiamento cl?nico, estadiamento patol?gico, envolvimento linfonodal, grau de diferencia??o histol?gico e sobrevida. Resultados: A express?o do FT foi observada em 26 esp?cimes (76,4%). A m?dia da express?o na amostra foi de 27,8 μm2 [IC 95% 20,0 35,6, DP 22,4]. Houve diferen?a significativa em rela??o a idade entre os grupos com express?o alta e baixa [57,6 anos (DP 9,9) contra 67,3 anos (DP 16,4), p = 0,047]. O seguimento m?dio foi de 28,3 meses (intervalo de 1 a 139,6 meses). Durante o acompanhamento houve 8 ?bitos (76,5%), 4 em cada grupo, todos relacionados a neoplasia. Na an?lise multivariada, n?o houve correla??o entre a express?o do FT e as vari?veis em estudo. Conclus?es: O FT foi expresso na maioria das amostras de c?ncer de p?nis. N?o houve correla??o entre n?vel de express?o do FT, fatores progn?sticos e sobrevida.

Made available in DSpace on 2015-04-14T13:35:58Z (GMT). No. of bitstreams: 1 458593.pdf: 1436191 bytes, checksum: b126309dee58688d2fe23cb45be67db7 (MD5) Previous issue date: 2014-02-21 / The purinergic system can induce proliferation, differentiation and death on tumor cells. Different P2 purinergic receptors are involved in preventing the growth of these cells. Although the mechanisms are not completely known, they may represent interesting therapeutic targets to control tumors. The RT4 and T24 human tumor bladder cell lines have been used as low and high cancer malignancy phenotype models, respectively. Our research group has demonstrated that quercetin increases the ADP hydrolysis and inhibits ecto-5'-nucleotidase/ CD73, exerting an anti-proliferative effect on T24 cell lines. We also demonstrate that T24 cell lines expresses the purinergic enzymes CD39L4 and ecto-5'-nucleotidase / CD73, and have low hydrolytic activity of ATPase and ADPase in contrast to a high activity AMPase. These data suggest the involvement of the enzymes and purinergic receptors on progression the tumor of the bladder, as well as other tumor types. The objective of the present study was characterize the purinergic receptors P2X (1-7) expression profile on human bladder tumor cell lines and biopsies; and investigate the inflammatory genes that are up-regulated on high-grade human tumor bladder cells compared to low grade tumor bladder cells. Human bladder cells RT4 and T24 were obtained from the ATCC. Cells were cultured in DMEN and RPMI, respectively, supplemented with 10% of fetal bovine serum (FBS), under ideal conditions of cultivation Data were analyzed by one-way analysis of variance (ANOVA), followed by Tukey Kramer test. To confirm the relevance of P2X (1-7) receptors in bladder cancer, we have also assessed their expression in human biopsies. For this purpose, upon approval by the local ethics committee, specimens of bladder tumors and normal tissues were obtained from the same patients, who have been undergone surgical resection at the Hospital S?o Lucas/PUCRS. All samples were collected and rapidly frozen in -80?C with RNA holder. Tissue specimens were ground and then sonicated in a TRIzol kit. The mRNA level was analyzed using RT-PCR analysis. Real-time PCR revealed that the mRNA expression of P2X purinergic receptors, particulary P2X4R was significantly up-regulated on both T24 and RT4 cell lines. Data from human bladder tumor biopsies comparing to bladder normal tissue, showed that P2X6 and P2X7 receptors appeared to be most often up-regulated on tumor tissues. Twelve inflammation related genes, CCL2, CCR2, TNF-α, MAPK3, IL-1β, IL-6, ADORA1, CD200, CD4, CHRNA4, EDNRA e ITGB2 were significantly up-regulated on the grade III bladder tumor cell line T24 when compared to grade I bladder tumor cell line RT4. Our results indicate that the inflammation genes overexpressed may play an important role in regulating urinary bladder cancer. We can conclude that P2X purinoreceptors and inflammation genes are differentially distributed among normal bladder, low and high tumor bladder, which could contribute to the different characteristics of these different types of cells. / O sistema purin?rgico pode induzir a prolifera??o, diferencia??o e morte em c?lulas tumorais. Diferentes receptores purin?rgicos P2, est?o envolvidos na inibi??o do crescimento tumoral. Embora os mecanismos n?o sejam completamente conhecidos, estes receptores podem representar alvos terap?uticos interessantes no tumor de bexiga. As linhagens celulares derivadas de tumor de bexiga humano RT4 e T24 t?m sido usadas como modelo de baixa e alta malignidade de tumor, respectivamente. Estudos realizados pelo nosso grupo de pesquisa t?m demonstrado o envolvimento do sistema purin?rgico no tumor de bexiga. A quercetina aumenta a hidr?lise do ADP e inibe a ecto - 5' - nucleotidase / CD73, exercendo um efeito anti - proliferativo sobre as c?lulas T24. Tamb?m demonstramos que as linhagens de c?lulas T24 expressam enzimas CD39L4 e ecto - 5' - nucleotidase / CD73, e t?m baixa atividade hidrol?tica de ATPase e ADP?sica com uma elevada atividade AMP?sica. Estes dados sugerem o envolvimento das enzimas e receptores purin?rgicos na progress?o do tumor de bexiga, bem como em outros tipos de tumor. O objetivo do estudo ? caracterizar a express?o dos receptores purin?rgicos P2X (1-7) em bi?psias de c?ncer de bexiga e linhagens celulares de tumor de bexiga humano, relacionado com o perfil de genes inflamat?rios mais expressos nestas amostras. Metodologia: As linhagens celulares de bexiga humanas RT4 e T24 foram obtidas a partir da ATCC. As c?lulas foram cultivadas em meio RPMI e DMEN, respectivamente, suplementado com 10 % de soro fetal bovino (SFB), sob condi??es ideais de cultivo. Para confirmar a relev?ncia dos receptores P2X (1-7) no tumor de bexiga, avaliou-se a express?o em bi?psias humanas. Para isso, ap?s aprova??o pelo comit? de ?tica local, amostras de tecidos normais e tumorais de bexiga foram obtidas dos mesmos pacientes, submetidos ? ressec??o cir?rgica no Hospital S?o Lucas / PUCRS. O RNA total dos tecidos foi isolado utilizando kit TRIzol . O n?vel de mRNA foi analisado utilizando an?lise RT - PCR . Resultados: O PCR em tempo real revelou que a express?o de mRNA de receptores purin?rgicos P2X, particularmente P2X4R foi significativamente up- regulada em ambas linhagens RT4 e T24. Dados a partir de bi?psias de tumores de bexiga humanos que comparam os resultados de express?o do tecido tumoral em rela??o ao tecido normal de bexiga, demonstrou que os receptores de P2X6 e P2X7 est?o superexpressos em tecidos tumorais. Doze genes relacionados com a inflama??o, CCL2, CCR2, TNF-α, MAPK3, IL-1β, IL-6, ADORA1, CD200, CD4, CHRNA4, EDNRA e ITGB2 foram significativamente superexpressos em c?lulas T24 derivadas de tumor de grau III quando comparadas com as c?lulas RT4 derivadas de tumor de grau I. Nossos resultados indicam que os genes superexpressos de inflama??o podem desempenhar um papel importante na regula??o do tumor de bexiga urin?ria. Conclus?o: Podemos concluir que purinoreceptores P2X e os genes inflamat?rios s?o diferencialmente distribu?dos entre bexiga normal e tumoral, o que pode contribuir para as diferentes caracter?sticas destes diferentes tipos de tumores.

Made available in DSpace on 2015-04-14T13:36:01Z (GMT). No. of bitstreams: 1 462235.pdf: 2683765 bytes, checksum: dc15bd33e02ceefcb7c6427763878df3 (MD5) Previous issue date: 2014-08-11 / It is unclear how temperament is related to alcohol-related behavior in large population studies. We have used the Affective and Emotional Composite Temperament Scale (AFECTS) model to evaluate how emotional traits and affective temperaments are associated with alcohol use, abuse, and dependence in the general population. Data from 10,603 subjects (mean age=28.0?7.8yrs, 70.3% females) was collected anonymously by the Internet in Brazil using the AFECTS model and the Alcohol, Smoking and Substance Involvement Screening Test (ASSIST). Alcohol use was stratified into control, low use, abuse, and dependence groups. The analysis of dimensional traits showed that Volition and Coping were lower, and Sensitivity was higher, in the abuse and dependence groups, with no differences between the Control and the Low Use groups. Alcohol consumption was also associated with lower Control, Stability, and Caution, and higher with Anger, Anxiety, and Desire, with significant differences between all groups. Regarding affective temperament types, alcohol abuse and dependence were associated with euphoric and cyclothymic temperaments in both genders, which was mirrored by a lower frequency of both euthymic and hyperthymictypes. Only hyperthymics were overrepresented in the Control group for both genders. A global dysfunction of emotional traits and a predominance of cyclothymic and euphoric temperaments were associated with alcohol-related behavior. Prevention and treatment strategies may be developed more effectively if these traits are taken into account. / N?o est? claro na literatura como otemperamento associa-se a comportamentos relacionados ao consumo de ?lcool em grandes estudos populacionais. Utilizamos o modelo Affective and Emotional Composite Temperament Scale (AFECTS) para avaliar como tra?os emocionais e temperamentos afetivos relacionam-se ao uso, abuso e depend?ncia de ?lcool na popula??o geral. Dados de 10.603 participantes (m?dia de idade = 28.0 ? 7.8 anos, 70,3% mulheres) foram coletados anonimamente pela Internet no Brasil utilizando o modelo AFECTS e o Alcohol, Smoking and Substance Involvement Screening Test (ASSIST). Estratificamos os sujeitos de acordo com o consumo da subst?ncia, sendo os grupos respectivamente nomeados: controle (baix?ssimo uso), baixo uso, abuso e depend?ncia. As an?lises para tra?os emocionais mostraram que nos grupos de abuso e depend?ncia os n?veis de Vontade e Maturidade (Coping) foram mais baixos, enquanto que os escores de Sensibilidade estavam mais altos, sem diferen?as entre os grupos controle e baixo uso. O consumo de ?lcool foi tamb?m associado com baixo Controle, Estabilidade e Cautela, alta Raiva, Ansiedade e Desejo, apresentando diferen?as significativas entre todos os grupos. Com rela??o aos tipos de temperamento afetivo, abuso e depend?ncia de ?lcool foram associados com temperamentos euf?rico e ciclot?mico em ambos os sexos, acompanhados de uma menor freq??ncia dos tipos eut?mico e hipert?mico. Apenas os hipert?micos tiveram alta representatividade no grupo controle, tanto para homens quanto para mulheres. A exist?ncia de uma disfun??o global de tra?os emocionais e a predomin?ncia de temperamentos ciclot?mico e euf?rico foram associados a comportamentos relacionados ao consumo de ?lcool. Ressalta-se que estrat?gias de preven??o e tratamento podem ser desenvolvidas de forma mais eficaz,sendo estas caracter?sticas identificadas e consideradas.

Made available in DSpace on 2015-04-14T13:36:05Z (GMT). No. of bitstreams: 1 390396.pdf: 742166 bytes, checksum: ff6f453c7552409274aceb3384b4bfd9 (MD5) Previous issue date: 2007-03-08 / Base te?rica: Atualmente, a avalia??o progn?stica do c?ncer de mama inicial ? principalmente baseada nas caracter?sticas cl?nicas das pacientes e histol?gicas dos tumores. Recentemente, subtipos moleculares de carcinoma invasor da mama foram reconhecidos atrav?s dos estudos de perfilamento gen?tico por DNA microarray. Essa nova classifica??o pode melhorar a avalia??o progn?stica, mas essa nova tecnologia ainda n?o ? amplamente dispon?vel. Assim, a possibilidade de identificar os subtipos moleculares atrav?s de m?todos mais simples e baratos ? promissora. A identifica??o do subtipo basal-like de c?ncer de mama particularmente importante porque tem uma evolu??o desfavor?vel e menos op??es de tratamento dispon?veis. Adicionalmente, a express?o de Receptor para o Fator de Crescimento Epid?rmico (EGFR) parece ser mais freq?ente nesse subtipo. M?todos: Uma coorte retrospectiva de 112 pacientes consecutivas com carcinoma prim?rio de mama de est?gio patol?gico I ou II, diagnosticadas e tratadas na mesma institui??o entre 1995 e 2000, foi estudada. As caracter?sticas histol?gicas e cl?nicas, bem como dados de evolu??o cl?nica e sobrevida, foram revisados. An?lise imunohistoqu?mica foi realizada em blocos representativos dos tumores com anticorpos contra Receptor de Estr?geno (RE), Receptor para o Fator de Crescimento Epid?rmico Humano-tipo 2 (HER2), CK 5/6 e EGFR. O objetivo prim?rio foi identificar a preval?ncia do subtipo basal-like de c?ncer de mama (RE HER2-negativo e CK 5/6 e/ou EGFR positivo) nessa popula??o. Resultados: 13 dos 112 (11,6%) tumores avaliados foram do subtipo basal-like. Sua idade m?dia foi 49 anos; 77 % eram est?gio patol?gico II e 100 % eram carcinomas ductais invasores. N?o houve diferen?a progn?stica entre o subtipo basal-like e os subtipos luminal (RE positivo e HER2 negativo); com superexpress?o de HER2 (HER2 positivo) e indeterminado (quatro marcadores negativos) com rela??o ? sobrevida livre de doen?a e sobrevida geral. Esse resultado provavelmente se deve ao n?mero limitado de pacientes neste estudo e ao bom progn?stico de pacientes com c?ncer de mama em est?gio inicial. 11 dos 112 (10%) casos foram positivos para EGFR. Tumores positivos para EGFR foram mais freq?entemente negativos para RE (23 x 5%;p=0,01). Conclus?es: Nossos resultados sugerem que ? poss?vel identificar o subtipo basal-like de c?ncer de mama atrav?s da an?lise imunohistoqu?mica do RE, HER2, CK 5/6 e EGFR. Al?m disso, a express?o de EGFR parece ser mais freq?ente em tumores Re-negativos, o que sugere que drogas direcionadas contra o EGFR poderiam beneficiar esse subgrupo de pacientes

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2016-07-13T20:15:38Z No. of bitstreams: 1 DIS_ANDREIA_PEREIRA_DE_FREITAS_COMPLETO.pdf: 1498465 bytes, checksum: 6d337a30869c0650d4c25ae34e767df0 (MD5) / Made available in DSpace on 2016-07-13T20:15:38Z (GMT). No. of bitstreams: 1 DIS_ANDREIA_PEREIRA_DE_FREITAS_COMPLETO.pdf: 1498465 bytes, checksum: 6d337a30869c0650d4c25ae34e767df0 (MD5) Previous issue date: 2016-03-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Funda??o de Amparo ? Pesquisa do Estado do Rio Grande do Sul - FAPERGS / The Brazilian economy adopts the regime of inflation targets since 1999 and one of the biggest challenges of monetary policy is to make the price takers form expectations and thus fix prices. Expectations guide the decisions of economic agents and are an important monetary policy transmission channel. Therefore, it is important that the monetary authority is transparent and has credibility with the public. An inflation target for a particular period is fully credible if it is equal to the inflation expectations of the market for the same period. This research aims to assess the way in which the expectations of economic agents influence the results in different economic indicators: Industrial Production, Exchange, Quarterly Agriculture GDP, Quarterly Industry GDP, Quarterly Service GDP, Quarterly Total GDP, IGPM, IPCA , Selic rate. The research analyzes how expectations affect the process of formation of own expectations and how the previous results influence the outcome of the next period and the formation of expectations. It uses the data of the statistical series available by Central Bank of Brazil of market expectations. The methodology is the time series and especially the Least Square Robust Models. We found the following results: For the Industrial Production and Exchange, previous expectations influenced the Last Expectation and the previous results were significant in the Result. For the IGP-M, only the previous results influenced the outcome, for the Last Expectation, all variables (previous expectations, previous results and deviations from previous expectations) were significant. For the IPCA and the Selic, all variables were statistically significant, the most relevant are short-term previous expectations, acting in the results and in the Last Expectation. For the Agriculture GDP, only previous expectations influenced the Last Expectation, and all variables had influence in the result. No variable searched for Industry GDP and Service GDP had an influence on the result. In Last Expectation, short-term expectations were significant. For the Total GDP, the previous expectations are significant in the result. Just previous expectations are significant in Average Last Expectation and only the expectation of two days before is significant in Median. / A economia brasileira adota o regime de metas de infla??o desde 1999 e um dos maiores desafios de uma pol?tica monet?ria ? fazer com que os tomadores de pre?os formem expectativas e assim fixem pre?os. As expectativas orientam as decis?es dos agentes econ?micos e s?o um importante canal de transmiss?o de pol?tica monet?ria. Sendo assim, ? importante que a Autoridade Monet?ria seja transparente e tenha credibilidade perante o p?blico. Uma meta de infla??o para um determinado per?odo ? plenamente cr?vel se ? igual ? expectativa de infla??o do mercado para o mesmo per?odo. Essa pesquisa busca avaliar a forma pela qual as expectativas dos agentes econ?micos influenciam os resultados em diferentes indicadores econ?micos: Produ??o Industrial, C?mbio, PIB trimestral agropecu?ria, PIB Trimestral Ind?stria, PIB Trimestral Servi?o, PIB Trimestral Total, IGPM, IPCA, Taxa Selic. A pesquisa analisa como as expectativas afetam o processo de forma??o das pr?prias expectativas e como os resultados anteriores influenciam no resultado do pr?ximo per?odo e na forma??o das expectativas. Utiliza os dados das s?ries estat?sticas, dispon?veis em Expectativas de Mercado divulgadas pelo Banco Central do Brasil. A metodologia ? a de S?ries Temporais e em especial os Modelos de M?nimos Quadrados Robustos. Encontramos como resultados: Para a Produ??o Industrial e C?mbio, as expectativas anteriores influenciaram a ?ltima Expectativa e os resultados anteriores foram significativos no Resultado. Para o IGPM, apenas os resultados anteriores influenciaram no resultado, j? na ?ltima Expectativa, todas as vari?veis (expectativas anteriores, resultados anteriores e desvios das expectativas anteriores) foram significativas. IPCA e Selic, tanto a atua??o no resultado como na ?ltima Expectativa, todas as vari?veis se mostraram significativas, sendo as mais relevantes ?s expectativas anteriores de curto prazo. Para o PIB da Agropecu?ria, apenas as expectativas anteriores tiveram influ?ncia na ?ltima Expectativa, e para o seu resultado, todas as vari?veis tiveram influ?ncia. Nenhuma vari?vel pesquisada para o PIB Ind?stria e o PIB Servi?o, teve influ?ncia no resultado, enquanto que na ?ltima Expectativa, as expectativas de curto prazo foram significativas. As expectativas anteriores s?o significativas para o resultado do PIB Total. J? para a M?dia da ?ltima Expectativa, apenas as expectativas anteriores foram significativas e na Mediana, somente a expectativa de 2 dias antes foi relevante.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2016-07-14T16:57:14Z No. of bitstreams: 1 DIS_LAURA_SMIDT_NUNES_PARCIAL.pdf: 228259 bytes, checksum: 74df0dbd4064a00e8b0071f07e6d7ecf (MD5) / Made available in DSpace on 2016-07-14T16:57:14Z (GMT). No. of bitstreams: 1 DIS_LAURA_SMIDT_NUNES_PARCIAL.pdf: 228259 bytes, checksum: 74df0dbd4064a00e8b0071f07e6d7ecf (MD5) Previous issue date: 2016-03-08 / Conselho Nacional de Pesquisa e Desenvolvimento Cient?fico e Tecnol?gico - CNPq / The aim of the study was to evaluate the surface roughness and analyze the surface topography of five different ceramics, a resin composite and two polymers for CAD/CAM after milling and post milling procedures. Blocks of Mark II, IPS Empress CAD, IPS e.max CAD, Enamic, Lava Ultimate, Suprinity, CAD-Temp and CAD-Waxx were milled in CEREC MCXL system. The surface roughness (Ra; ?m) was measured before and after millig procedures, polishing and glaze (when indicated) followed by analysis by SEM and AFM. The results were subjected to statistical tests (one-way ANOVA for repeated measures and Tukey HSD and one-way ANOVA also with Post-Hoc Tukey, respectively) with a significance level of 5% (?=0.05). The milling step significantly increased surface roughness in all tested materials (p<0.05). Mechanical polishing was able to reduce the roughness generated by grinding, requiring the glazing step only in partially crystallized ceramic (IPS e.max CAD and Suprinity). In other materials (Mark II, IPS Empress CAD, Lava Ultimate, Enamic, CAD- Temp and CAD-Waxx) mechanical polishing was enough to achieve similar surface roughness values of the smoothness of the blocks directly from manufacturers. The study suggests that the glaze step can be suppressed in materials completely crystallized and/or polymerized by the manufacturer, these having no statistically significant difference when it's just done mechanical polishing. / O objetivo do estudo foi avaliar a rugosidade superficial e analisar a topografia de superf?cie de cinco diferentes cer?micas, um comp?sito resinoso e dois acr?licos para CAD/CAM ap?s usinagem e procedimentos p?s-usinagem. Blocos de Mark II, IPS Empress CAD, IPS e.max CAD, Enamic, Lava Ultimate, Suprinity, CAD-Temp e CAD- Waxx foram usinados no sistema CEREC MCXL. A rugosidade superficial (Ra; ?m) foi aferida antes e ap?s os procedimentos de usinagem, polimento e glaze (quando indicado) seguidos de an?lises em MEV e MFA. Os resultados foram submetidos a testes estat?sticos (teste one-way ANOVA para medidas repetidas e Tukey HSD e ANOVA one-way tamb?m com Post-Hoc Tukey, respectivamente) com n?vel de signific?ncia de 5% (?=0,05). A etapa de fresagem aumentou significativamente a rugosidade superficial em todos os materiais testados (p<0,05). O polimento foi capaz de reduzir a rugosidade gerada pela usinagem, sendo necess?ria a etapa de glazeamento apenas para cer?micas parcialmente cristalizadas (IPS e.max CAD e Suprinity). Nos demais materiais (Mark II, IPS Empress CAD, Lava Ultimate, Enamic, CAD-Temp e CAD-Waxx) o polimento mec?nico foi suficiente para atingir valores de rugosidade superficial similares a lisura dos blocos provenientes diretamente dos fabricantes. O estudo sugere que a etapa de glaze pode ser suprimida nos materiais j? provenientes do fabricante completamente cristalizados e/ou polimerizados, nestes n?o possuindo diferen?a estatisticamente significativa quando apenas ? realizado polimento mec?nico.