Germline mutations in mismatch repair genes predispose individuals to Lynch Syndrome, the most common colorectal cancer predisposition syndrome. MLH1 is a key mismatch repair gene that is mutated in Lynch syndrome and about a third of the genetic alterations identified in MLH1 are missense variants of unclassified clinical significance. We hypothesize that missense alterations in distinct domains of MLH1 likely affect its expression and function(s) to varying degrees. To address this we utilized several approaches to investigate the molecular basis of the pathogenicity of a panel of unclassified variants. Our results demonstrate that the MLH1 variants p.R265C and p.K618A significantly decrease the stability of the MLH1 protein, while the variant p.L749Q compromises heterodimerization of the MLH1-PMS2 complex. Given the limitations and complexity of in vitro assessment strategies, we conducted a proof-of-principle study to investigate whether missense variants in MLH1 lead to allelic imbalances at the transcriptional level. Our analysis using the PeakPicker software indicated that the missense variants c.350C>T, c.793C>T, c.1852_1853AA>GC, as well as the truncating variant c.1528C>T were associated with significantly unbalanced allelic expression. This illustrates a novel method of investigating the pathogenicity of unclassified genetic variants, which has the potential to be applied in the diagnostic setting. Previous genetic epidemiology studies from our laboratory have demonstrated that the MLH1 c.-93G>A promoter variant is strongly associated with the microsatellite instability phenotype in colorectal tumours. Additionally, this promoter variant was associated with an elevated risk of endometrial cancer in case-control studies. Results from our functional studies indicate that the c.-93G>A variant significantly alters the promoter activity of MLH1. The MLH1 promoter is bi-directional with the EPM2AIP1 gene located on the antisense strand. Interestingly, we observed that this variant significantly affected EPM2AIP1 transcription as well. Furthermore, our experiments suggest that c.-93G>A variant affects transcription by altering the affinity of nuclear factors that bind this region. Combined, these findings shed light on the possible mechanisms by which missense variants affect MLH1 expression and function, which in conjunction with results from other functional assays will help increase the accuracy and efficiency of genetic testing of inherited cancers.

Germline mutations in mismatch repair genes predispose individuals to Lynch Syndrome, the most common colorectal cancer predisposition syndrome. MLH1 is a key mismatch repair gene that is mutated in Lynch syndrome and about a third of the genetic alterations identified in MLH1 are missense variants of unclassified clinical significance. We hypothesize that missense alterations in distinct domains of MLH1 likely affect its expression and function(s) to varying degrees. To address this we utilized several approaches to investigate the molecular basis of the pathogenicity of a panel of unclassified variants. Our results demonstrate that the MLH1 variants p.R265C and p.K618A significantly decrease the stability of the MLH1 protein, while the variant p.L749Q compromises heterodimerization of the MLH1-PMS2 complex. Given the limitations and complexity of in vitro assessment strategies, we conducted a proof-of-principle study to investigate whether missense variants in MLH1 lead to allelic imbalances at the transcriptional level. Our analysis using the PeakPicker software indicated that the missense variants c.350C>T, c.793C>T, c.1852_1853AA>GC, as well as the truncating variant c.1528C>T were associated with significantly unbalanced allelic expression. This illustrates a novel method of investigating the pathogenicity of unclassified genetic variants, which has the potential to be applied in the diagnostic setting. Previous genetic epidemiology studies from our laboratory have demonstrated that the MLH1 c.-93G>A promoter variant is strongly associated with the microsatellite instability phenotype in colorectal tumours. Additionally, this promoter variant was associated with an elevated risk of endometrial cancer in case-control studies. Results from our functional studies indicate that the c.-93G>A variant significantly alters the promoter activity of MLH1. The MLH1 promoter is bi-directional with the EPM2AIP1 gene located on the antisense strand. Interestingly, we observed that this variant significantly affected EPM2AIP1 transcription as well. Furthermore, our experiments suggest that c.-93G>A variant affects transcription by altering the affinity of nuclear factors that bind this region. Combined, these findings shed light on the possible mechanisms by which missense variants affect MLH1 expression and function, which in conjunction with results from other functional assays will help increase the accuracy and efficiency of genetic testing of inherited cancers.

Thesis (Ph. D.)--University of Texas at Austin, 2007. / Vita. Includes bibliographical references.

Submitted by Paula Coutinho (paulaacoutinho@yahoo.com.br) on 2018-02-20T17:16:51Z No. of bitstreams: 1 Dissertação [Paula Andrade Coutinho] - PPGMUSEU - UFBA.pdf: 4747717 bytes, checksum: 0706070a9d518b2d6207b23ef342588e (MD5) / Approved for entry into archive by Biblioteca Isaías Alves (reposiufbat@hotmail.com) on 2018-02-21T14:00:04Z (GMT) No. of bitstreams: 1 Dissertação [Paula Andrade Coutinho] - PPGMUSEU - UFBA.pdf: 4747717 bytes, checksum: 0706070a9d518b2d6207b23ef342588e (MD5) / Made available in DSpace on 2018-02-21T14:00:04Z (GMT). No. of bitstreams: 1 Dissertação [Paula Andrade Coutinho] - PPGMUSEU - UFBA.pdf: 4747717 bytes, checksum: 0706070a9d518b2d6207b23ef342588e (MD5) / A presente dissertação objetiva apresentar a trajetória do colecionador carioca Henry Joseph Lynch por meio de seu colecionismo. Procuramos compreender o processo de fabricação de sua imagem a partir das estratégias que mobilizou para consagrar-se socialmente. A construção do seu autorretrato social encontrou amparo em sua coleção-imagem, veículo de distinção social e aquisição de poder simbólico. Lynch procurou legitimar-se em vida e perpetuar o seu nome, imortalizar-se. Para isso, garantiu a preservação de seu legado ao doar, via testamento, o conjunto à Sociedade Brasileira de Cultura Inglesa (SBCI). Esta instituição distingue o conjunto em duas coleções: a Coleção Cultura Inglesa, correspondente à pinacoteca, e a Coleção Sir Henry Lynch, constituída por sua biblioteca. Posteriormente, há o desmembramento das duas coleções. No ano de 2000, a Coleção Cultura Inglesa é adquirida pelo colecionador pernambucano Ricardo Brennand para compor o acervo do Instituto Ricardo Brennand (Instituto RB). Na trajetória da coleção, novos significados são construídos. Na SBCI verificamos a manutenção da memória do colecionador, ao passo em que a instituição promovia sua própria imagem. No Instituto RB a coleção adquire novas configurações e, consequentemente, diversificam-se os significados, que passam a remeter, então, mais ao colecionismo de Ricardo Brennand do que ao de Henry Lynch. Na condição atual do conjunto custodiado pelo Instituto RB, refletimos, em paralelo, sobre as possibilidades de construção de novas narrativas, com base na documentação, lançando luz às procedências, a partir das quais os guardiões ganham visibilidade. / The aim of this dissertation is to present the trajectory of Rio-based collector Henry Joseph Lynch through his collectionism. Using the strategies, he assembled in order to establish himself socially as our starting point, we try to understand the process through which he created his public image. The elaboration of his social self-portrait found support in his collection, the means through which he distinguished himself socially and acquired symbolic power. Lynch sought to legitimize himself while alive and to perpetuate his name, or rather, to immortalize himself. It was with this intent that he secured his legacy, by donating the entirety of his collection to the Brazilian Society of English Culture (SBCI). This institution organized said collection into two segments: the English Culture Collection, which corresponds to the art gallery, and the Sir Henry Lynch Collection, which corresponds to his library. Subsequently, the two collections were separated. In 2000, the English Culture Collection was acquired by Pernambuco collector Ricardo Brennand, who incorporated it to the permanent collection of the Ricardo Brennand Institute (IRB). New meanings are acquired throughout the trajectory of the collection. On SBCI, we verify the preservation of the memory of the collector, as the institution promoted its own image alongside it. On the IRB, the collection is reorganized, and consequently it acquires new meanings, which relate more to the collectionism of Ricardo Brennand than to that of Henry Lynch. According to the present condition of the collection currently held by the IRB, we reflect upon the possibility of the elaboration of new narratives based on the related documentation, shedding a light on the provenance of the art works therein, and according to which its guardians become more visible.

Orientador: Carmen Sílvia Bertuzzo / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-23T00:44:00Z (GMT). No. of bitstreams: 1 Rueda_LidianeCamila_D.pdf: 2777204 bytes, checksum: dc2eb94ba9de0c91e1c7a3fed2fbd3e4 (MD5) Previous issue date: 2013 / Resumo: O câncer colorretal tem importância elevada frente a sua incidência e morbidade. Dentre os casos hereditários, o câncer colorretal hereditário sem polipose (HNPCC), ou Síndrome de Lynch, é responsável por cerca de 5% do total de casos. No HNPCC, a alteração genética herdada é a inativação de um dos alelos dos genes envolvidos em reparo do DNA, sendo os principais os genes hMLH1 e hMSH2. O objetivo deste trabalho foi investigar, em indivíduos com diagnóstico clínico de HNPCC, a presença de mutações nos genes MLH1 e MSH2, associar as variáveis clínicas com o gene mutado e investigar os familiares de portadores de HNPCC aos quais tivemos acesso, com relação a mutações germinativas. A investigação das mutações foi realizada por meio de sequenciamento direto dos éxons, região promotora e regiões de junção. Foram analisados 65 indivíduos divididos em três grupos, sendo (I) 46 pacientes portadores de câncer colorretal inclusos nos Critérios de Amsterdã, (II) dois familiares portadores de câncer colorretal e (III) 17 familiares sem câncer, todos da região metropolitana de Campinas, atendidos no Hospital de Clínicas da UNICAMP. Em 21 (45,65%) dos pacientes foram encontradas mutações deletérias. As mutações deletérias nos genes MLH1 e MSH2 estavam na proporção de 34,78% (16 pacientes) e 10,86% (5 pacientes), respectivamente. As mutações não deletérias nos genes MLH1 e MSH2 estavam na proporção de 65,22% dos pacientes (30 alterações) e 50% dos pacientes (23 alterações), respectivamente. Foi possível identificar 23 mutações potencialmente deletérias entre os pacientes com HNPCC por meio de sequenciamento dos genes MLH1 e MSH2, com uma porcentagem de detecção de 50%. Parece não haver variações nas características clínicas do tumor quando a mutação germinativa ocorre no gene MLH1 ou MSH2, com exceção da relação entre presença de mutação no gene MLH1 e idade de manifestação da doença. Como ocorre no resto do mundo a doença mostrou-se extremamente heterogênea em termos moleculares, pois apenas duas mutações se repetiram em dois pacientes. A partir da análise das duas famílias foi possível mostrar a dificuldade para estabelecer a presença da mutação germinativa deletéria que poderia levar à predisposição ao HNPCC, bem como a importância da analise familial no diagnóstico molecular dessa alteração / Abstract: Colorectal cancer has high importance because of its incidence and morbidity. Among the hereditary cases, the hereditary nonpolyposis colorectal cancer (HNPCC) or Lynch syndrome, accounts for about 5% of cases. In HNPCC, the genetic alteration inherited is the inactivation of one of the alleles of genes involved in the DNA repair, being hMSH2 and hMLH1 the main genes. The objective of this study is to investigate the presence of mutations in MLH1 and MSH2 in patients with clinical diagnosis of HNPCC, correlate clinical variables with the mutated gene, and investigate the relatives of patients with HNPCC who we had access to, in relation to germline mutations. Investigation of the mutations was performed by éxons direct sequencing, the promoter and junction regions. Sixty-five individuals, divided into three groups, were studied: (I) 46 patients with colorectal cancer included in the Amsterdam Criteria, (II) two family members of colorectal cancer patients and (III) 17 relatives without cancer, all of them treated at Hospital das Clínicas at UNICAMP and living in the Campinas metropolitan area. Deleterious mutations were found in 21 patients (45.65%). The ratio of deleterious mutations in MLH1 and MSH2 was 34.78% (16 patients) and 10.86% (5 patients) respectively. The ratio of non deleterious mutations in genes MLH1 and MSH2 was 65.22% of patients (30 alterations) and 50% of patients (23 alterations) respectively. Among patients with HNPCC, 23 potentially deleterious mutations were identified, via sequences of MLH1 and MSH2 with a 50% detection rate. It doesn't seem to appear variations in the clinical characteristics of the tumor when a germline mutation occurs in MLH1 or MSH2, with the exception of the relationship between the presence of mutation in the MLH1 gene and age of disease onset. As it occurs throughout the world, the disease present a his molecular extremely heterogeneoty, where only two mutations were repeated in two patients. The analysis of the two families demonstrated not only the difficulty to establish the presence of deleterious germline mutation that could lead to a predisposition to of HNPCC, but also the importance of familial analysis in molecular diagnostics of this alteration / Doutorado / Clinica Medica / Doutora em Ciências

Aims: The purpose of the research was to investigate the benefits of a hereditary colorectal cancer registry in the management of patients and families with Lynch syndrome. In study one, a systematic review was performed to quantify the impact of registration and screening on colorectal cancer (CRC) incidence and mortality, with comparison between familial adenomatous polyposis (FAP) and Lynch syndrome (LS). In study two, a regional Lynch syndrome registry was utilised to evaluate the uptake of predictive testing and colorectal screening among first-degree relatives (FDRs) and investigate novel methods for engaging at-risk relatives, including an enhanced role for the general practitioner (GP). In study three, the registry was used to investigate proposed associations between Lynch syndrome and prostate and bladder cancer. In study four, mismatch repair-deficient (dMMR) CRCs from Lynch syndrome patients and randomised-controlled trials (RCTs) were used to evaluate a novel prognostic biomarker, beta-2 microglobulin (B2M). Methods: An electronic database search was conducted to identify studies describing CRC incidence and/or mortality in FAP or LS, with comparison of either: 1) screened and unscreened patients or 2) patients ‘before and after’ establishment of the registry. Using the Manchester regional Lynch syndrome registry database, the uptake of predictive testing and colorectal screening among FDRs was assessed with Kaplan-Meier analysis. Novel strategies for improving engagement were explored via a patient advisory group discussion and a regional primary care questionnaire. Cases of prostate and bladder cancer in male mutation carriers and their male FDRs were identified, and cumulative and relative risks were calculated, using expected rates from cancer registry data. DNA from 350 dMMR CRC specimens from Lynch syndrome patients and RCTs were tested for B2M mutations using Sanger sequencing, and correlated with clinical outcome. Results: 43 studies were included in the systematic review (33 FAP; 10 Lynch). Registry-based screening was associated with a significant reduction in CRC incidence and in Lynch syndrome, CRC-related mortality was negligible in those undergoing surveillance. 242 Lynch syndrome families were recorded on the Manchester Lynch syndrome registry. 329 of 591 (55.7%) eligible FDRs had undergone predictive testing. Uptake was significantly lower in males and younger age groups (<25 yrs). Compliance with colorectal screening was excellent following a mutation positive predictive test but poor in untested individuals (97.3% vs 35.0%). Eight prostate cancers were identified in 821 male LS mutation carriers and male FDRs. MSH2 mutation carriers had a ten-fold increased risk of prostate cancer (RR 10.41; 95%CI 2.80, 26.65) but no association with bladder cancer was identified. 69/286 (24.1%) of dMMR CRCs contained significant B2M mutations. B2M mutations were associated with complete absence of recurrence (0/39) during follow-up in the QUASAR trial (stage II), compared with 14/77 (18.2%) in wild-type B2M (p=0.005). Conclusion: Studies consistently report that registration and screening result in a reduction of CRC incidence and mortality in FAP and LS (Level 2a evidence, Grade B recommendation). Funding and managerial support for registries should be made available. Uptake of predictive testing and colorectal screening in Lynch syndrome could be substantially improved, particularly among males and younger age groups, but this requires advances in communication with at-risk relatives. It is unlikely that GPs will actively participate without considerable support from genetics services. A trial of PSA screening in MSH2 mutation carriers from 50 years would be appropriate. B2M mutation status has potential clinical utility as a prognostic biomarker in stage II dMMR CRC.

Der Artikel untersucht einen Werbespot von David Lynch für die Playstation 2 und ordnet ihn in das Universum der Lynch’schen Texte, das Lynchville, ein. Zunächst werden dazu einige regelmäßig wiederkehrende Motive aus dem Werk Lynchs vorgestellt. Es folgt eine eingehende Analyse des Werbespots, in der zahlreiche Parallelen zu den bekannten Filmen des Œuvres aufgezeigt werden. Abschließend wird kurz der Frage nachgegangen, inwiefern der Begriff „auteur“ in dieser wohl kommerziellsten aller filmischen Ausdrucksformen überhaupt anwendbar ist. Der Artikel kommt zu dem Ergebnis, dass es Lynch in diesem Werbespot gelingt, eine persönliche Visiondurchscheinen zu lassen – und dass die Karte des Lynchville deshalb um einen neuen Ort ergänzt werden muss. / The article analyzes a television-commercial, which David Lynch shot for the launch of Playstation 2. It tries to introduce that commercial into the universe of well known Lynch-Movies, the Lynchville. At first, some of the recurrent motives in Lynch’s work will be discussed. Then the commercial will be analyzed, especially regarding any parallels to Lynch’s Œuvre. Finally, the question will shortly be discussed whether in this most commercial form of filmmaking the term “auteur” can still apply. The article comes to the conclusion, that Lynch succeeds in maintaining his personal vision – and that therefore the map of Lynchville has to be expanded.

Background. Lynch syndrome is the most common genetic predisposition for hereditary cancer. Carriers of pathogenic changes in mismatch repair (MMR) genes have an increased risk of developing colorectal (CRC), endometrial, ovarian, urinary tract, prostate, and other cancers, depending on which gene is malfunctioning. In Lynch syndrome, differences in cancer incidence (penetrance) according to the gene involved have led to the stratification of cancer surveillance. By contrast, any differences in penetrance determined by the type of pathogenic variant remain unknown. Objective. To determine cumulative incidences of cancer in carriers of truncating and missense or aberrant splicing pathogenic variants of the MLH1 and MSH2 genes. Methods. Carriers of pathogenic variants of MLH1 (path_MLH1) and MSH2 (path_MSH2) genes filed in the Prospective Lynch Syndrome Database (PLSD) were categorized as truncating or missense/aberrant splicing according to the InSiGHT criteria for pathogenicity. Results. Among 5199 carriers, 1045 had missense or aberrant splicing variants, and 3930 had truncating variants. Prospective observation years for the two groups were 8205 and 34,141 years, respectively, after which there were no significant differences in incidences for cancer overall or for colorectal cancer or endometrial cancers separately. Conclusion. Truncating and missense or aberrant splicing pathogenic variants were associated with similar average cumulative incidences of cancer in carriers of path MLH1 and path_MSH2.

No description available.

我國綜合券商近年來因為整體家數過多、全球景氣持續衰退、金控時代來臨等因素影響下，使得整體產業經營出現困難，極待政府與民間共思一個解決方案。為此，財政部與證期會共同協助我國綜合券商的轉型升級，希望透過政府相關法令政策的開放解套，推動獨立綜合券商轉型為投資銀行，解決目前綜合券商在經營上所面臨之困境。 本研究針對我國綜合券商轉型投資銀行的相關政策，進行可行性研究與個案分析，並以國外知名投資銀行成功案例進行個案分析，從中探討投資銀行產業的成功關鍵因素，以做為我國綜合券商轉型投資銀行的策略擬定依據。 根據本研究成果，提出以下幾點結論： 1.台灣綜合券商面臨生存與轉型之壓力 2.我國整體投資環境確實有發展投資銀行可能性存在 3.產品創新、資本規模、品牌知名度及全球佈局能力仍是關鍵 4.各券商必須依據自身核心優勢差異發展不同之轉型策略 5.政府法令開放程度影響投資銀行發展 除此之外，本研究亦提出以下幾點建議： 1.對政府與相關主管單位建議：(1)法令開放持續推動；(2)建立有效監理機制；(3)大陸市場審慎評估。 2.對整體證券產業建議：(1)適度整合市場競爭狀況；(2)提供建議與政府合作。 3.對於個案公司建議：建議個案公司仔細思考目前所面臨轉型挑戰，以及相關解決辦法，透過本身核心優勢維持與發揮，逐步轉型投資銀行以因應我國金融產業轉型與升級。