MicroRNAs (miRNAs) are non-coding small RNAs that regulate eukaryotic gene activity at the post-transcriptional level by a process termed miRNA gene suppression. MicroRNA-122 (miR-122) is predominantly expressed in human liver cells and recent studies indicated that miR-122 promotes Hepatitis C Virus (HCV) replication and translation through physical interaction with two tandem binding sites located in the 5 untranslated region (5UTR) of the HCV genome (Jopling, et al., 2006; Jopling, et al., 2008). It has been reported that host genes that are also implicated in the miRNA gene suppression pathway are key regulators of HCV replication (Randall, et al., 2007). Two proteins, Dicer, a key RNaseIII enzyme, and its binding partner TRBP are essential proteins for miRNA activity. They are part of a protein complex called the RNA induced silencing complex (RISC) which also includes Argonaute proteins, and function in miRNA biogenesis loading the miRNA into RISC. As such, they are intriguing targets to study host-viral interplay during HCV replication.<p> In our study, we designed siRNAs to knock down Dicer and TRBP and then observed the effects of gene knockdown on full length J6/JFH-1-RLuc HCV (genotype 2a chimeric genome) replication and translation. The results showed that knocking down Dicer and TRBP reduced wild type (wt) J6/JFH-1-RLuc replication but had almost no effects on HCV translation in human liver cells. However, since knocking down Dicer and TRBP did not significantly alter miR-122 levels in the cell, it appears that the role of Dicer and TRBP was not solely the biogenesis of miR-122. This was confirmed by an experiment in which we observed that knocking down Dicer and TRBP also attenuated replication of a mutant virus in which replication is dependent on a exogenously supplied miRNA instead of endogenous miR-122.<p> Taken together, the results supported the hypotheses that Dicer and TRBP facilitate HCV infection mainly through HCV replication but not translation. The effects of Dicer and TRBP on HCV replication are not solely due to miR-122 biogenesis, and may be due to RISC loading functions in steps of miRNA gene suppression.<p> This study has set some essential groundwork for investigating potential roles of host factors in the RNAi machinery modulating HCV replication/translation and exploring novel antiviral targets.

It is difficult to ascertain the reason why the Lives of Suetonius have been so neglected by English-speaking scholars. In the historical significance of the period they cover, in the light they throw upon Roman life and manners and as a rich mine of anecdotes concerning Roman emperors, they have always been regarded as a most important source of information. There have been, however, few commentaries devoted to the Lives. The Galba has been particularly disregarded. Not since Mooney's edition of 1930 has there been any complete English commentary on the Life. However, other factors than the lack of a recent edition contributed to the choice of the Galba as the subject of this thesis. The Life deals with one of the most remarkable periods of Roman history, with the decline of the Julio-Claudian line and the subsequent political upheaval. It is also typical of Suetonius' style of biography in its impartiality, the structure and organization of its narrative, and its linguistic style. Finally, in the parallel accounts of Galba's life by Tacitus, Dio and Plutarch there exist constant points of reference that are vital for reasons of characterisation and historical value. It has been my aim in this thesis to produce a critical commentary on the Galba that both demonstrates the peculiar characteristics of the work and evaluates the contributions of modern scholarship. The thesis falls into three parts. The first chapter deals with the life of Suetonius and includes a discussion of the problems caused by the discovery of the Hippo Inscription. Subsequent sections are devoted to the date of composition of the Lives, the contentious question of Suetonius' sources and, finally, the manuscript tradition. Chapter two consists of a text of the Galba based on that of Ihm ( 1958). Some variations have been made and defended in the Commentary. Chapter three, the Commentary, constitutes the bulk of the thesis and is a section-by-section discussion of historical, textual and linguistic points arising from the text. The method of citation, through out, is by name of author and year of publication only. Full details can be found in the bibliography at the end of the thesis. / Arts, Faculty of / Classical, Near Eastern and Religious Studies, Department of / Graduate

Hepatitis C virus (HCV) is a serious global health problem with an estimate of 170 million carriers worldwide. Most individuals exposed to this blood-borne pathogen develop chronic infection, which may result in severe liver complications as well as end-stage liver diseases including cirrhosis and hepatocellular carcinoma. Current treatment options are suboptimal with no effective vaccines available to date. Development of a readily accessible mouse model that is permissive to natural HCV infection is important to facilitate drug and vaccine discovery, and also to better understand the viral pathogenesis. The inherent difficulty is that HCV displays very limited tropism, infecting only livers from humans or chimpanzees. An attempt was made to elucidate the key determinants in rendering the murine intracellular environment permissive to HCV replication. The results revealed that deletion of the interferon regulatory factor-3 and overexpression of microRNA-122 can independently enhance viral subgenomic replication in murine fibroblasts, with microRNA-122 being the stronger determinant. Interestingly, the phenotype established by these genetic manipulations was insufficient to support full-length HCV genome replication. Murine hepatic cell lines, with or without microRNA-122 expression, were also non-permissive to genomic HCV replication, despite the fact that translation of viral RNA was observed. These results suggest that additional host-specific factor(s) are required to support replication of full-length HCV RNA. These studies provide insight on the essential factors capable of influencing permissiveness of rodent cells to HCV replication, and also suggest genetic modifications to be considered when modeling the complete viral life cycle in a rodent animal model.

The aim of these studies was to identify circulating miRNAs that can be used as biomarkers in patients with paracetamol-induced liver injury. Whether the miRNAs discovered in humans could be back-translated to zebrafish with the aim of developing a liver toxicity model to replace rodent use was also investigated. First, the miRNA signature of DILI induced by paracetamol was defined. Plasma miRNAs were quantified in paracetamol overdose patients. A signature of 16 miRNAs was discovered that best separated patients with liver injury from those without liver injury. This signature was tested in a second cohort and resulted in the detection of paracetamol-induced liver injury with high specificity and sensitivity. At first presentation to hospital miR-122-5p was the most sensitive single miRNA and superior to ALT activity in predicting liver injury. In order to further qualify miR-122-5p, three detailed studies relevant to possible clinical scenarios were performed. The effect of acute alcohol ingestion (commonly co-ingested with paracetamol overdose) on circulating concentrations of miR-122-5p in healthy volunteers was investigated. Alcohol ingestion induced a small, non-clinically relevant, increase in miR-122-5p. The effect of chronic kidney disease (CKD) and haemodialysis (HD) on circulating miR-122-5p concentrations was explored because kidney dysfunction has been associated with a reduction in the concentration of circulating miRNAs. HD patients had lower concentrations of miR- 122-5p compared to healthy volunteers and CKD patients. To facilitate miRNA measurement outwith hospitals, miR-122-5p was measured in a blood drop from a finger prick. miR-122-5p was readily measurable in finger prick samples and concentrations were significantly higher in the blood drop from DILI patients compared with healthy volunteers. To complement miR-122-5p as a marker of toxicity, circulating paracetamol metabolites were measured in plasma samples from paracetamol overdose patients. A higher percentage of circulating metabolites formed by cytochrome P450 enzymes were present in patients with liver injury and these metabolites were superior to both ALT and paracetamol concentration with regard to early patient stratification. To reduce need for rodent studies, miRNAs were back-translated into zebrafish. In order to study circulating miR-122-5p in adult zebrafish, a bloodletting method by collecting blood retro-orbitally was developed. After studying different dosing regimens of paracetamol in adult and larvae zebrafish the model was determined to be too variable with regard to liver injury. A new drug, triptolide, originating from traditional Chinese medicine and responsible for DILI in China, was tested as an alternative model for drug-induced liver injury in zebrafish larvae. miRNA-122-5p decreased in zebrafish larvae after triptolide treatment and triptolide-induced liver injury could be tracked by fluorescent microscopy. Selective plane illumination microscopy was able to track the decrease in liver volume during triptolide exposure. In order to identify the toxic pathways involved in triptolide-induced liver injury, RNA-sequencing was performed. This identified KEGG pathways including ribosome, spliceosome and notch signalling as pathways affected by triptolide. In summary, miRNAs can be used as highly sensitive biomarkers to detect acute liver injury in patients and zebrafish. Zebrafish may represent an alternative model species to study DILI, further work is needed.

161 miospore species and 122 microplankton species are described from 106 samples in six sections of the Lower Colorado Group and its equivalents in central and west-central Alberta. Two miospore species, and two genera and twenty-five species of microplankton, are newly described. The miospore and microplankton assemblages are used as a basis for division of the Lower Colorado Group into several time-stratigraphic units. The age of the Group is determined as Upper Albian (latest Lower Cretaceous). A method for defining recurrent microplankton species groups is developed and applied to assemblages from two of the sections. The distribution of the recurrent groups formulated is shown to be correlated in part with palynological, lithological and other data. Application and extension of the results and conclusions of this investigation should prove useful in understanding many aspects of western Canadian Cretaceous stratigraphy and in the investigation of microplankton occurrence patterns from other deposits. / Thesis / Doctor of Philosophy (PhD)

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No description available.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / O PLC 122/2006 configurou um projeto de lei que visava criminalizar atos de preconceito e discriminação motivados por gênero, sexo, orientação sexual, identidade de gênero, idade e deficiência humana, alterando a Lei n 7.716, de 5 de janeiro de 1989, que criminaliza o preconceito motivado por raça, cor, etnia, religião ou procedência nacional. Desde perspectivas teológicas, a dissertação apresenta o PLC 122 a partir do seu nascedouro na Câmara dos Deputados em 2001, até a culminância do apensamento do PLC 122/2006 ao Projeto de Lei do Senado que intenta a reforma do Código Penal (PLS 236/2012) em 2013. No primeiro capítulo, foram elencados dados estatísticos e conceituais sobre a homofobia, programas governamentais que buscam a erradicação da homofobia no Brasil e no mundo, bem como uma contextualização histórica do PLC 122/2006. No segundo capítulo, elaborou-se um levantamento discursivo em torno da posição de grupos religiosos que se revelaram contrários ao PLC 122/2006, a saber, a Bancada Evangélica, a Conferência Nacional dos Bispos do Brasil (CNBB) e a Associação Brasileira de Instituições Educacionais Evangélicas (ABIEE). Essas vozes rebateram o PLC 122/2006 com argumentos que giraram em torno de uma possível restrição da liberdade religiosa caso o PLC fosse aprovado, o que gerou um conflito de direitos entre as partes que lutavam pró ou contra o projeto. A seguir, ilustrou-se o estudo com um breve levantamento acerca do discurso oficial e posicionamentos de outras vozes religiosas (sobretudo protestantes) que têm se pronunciando frente ao tema. No terceiro capítulo, abordou-se de que forma o embate exposto conversa com o estabelecimento da laicidade do Estado e a construção dos direitos humanos, identificando um conflito de direitos gerado a partir da denúncia de que a intervenção de vozes religiosas na política e na implementação das políticas públicas fere a noção de Estado laico. Em contrapartida, para os evangélicos, grupo que explicitamente se ergue em defesa da "moral cristã e da família normativa de um país de maioria cristã, esse rechaço fere, igualmente, a democracia e a constituição brasileira (1988), que assegura a todo/a cidadão/ã a liberdade religiosa e de expressão. Em direito outorgado pela constituição e em missão evocada pela confissão de fé cristã que ensina a defender a vida em sua diversidade acima de qualquer lei ou dogma, outras vozes eclesiais, a(s) teologia(s) pública(s) cristã(s) e as entidades ecumênicas são participantes públicas a contribuir para com o debate e a luta por direitos humanos das pessoas LGBT. Ao fim e ao cabo, defende-se o contínuo diálogo sobre a urgência em criminalizar a homofobia no Brasil, assumindo que a labuta é árdua, conflituosa e até dolorosa, mas o silêncio que oprime precisa ser quebrado e denunciado diariamente pela sociedade civil, pelas igrejas (em sua diversidade), teologias públicas e entidades ecumênicas e, não por último, pelos/as representantes do povo no Congresso Nacional para uma construção humana dos direitos humanos LGBT. / PLC 122/2006 was a bill aimed at criminalizing acts of prejudice and discrimination motivated by gender, sex, sexual orientation, gender identity, age and human disability, amending Law No. 7.716, of January 5, 1989, which criminalizes bias motivated by race, color, ethnicity, religion or national origin. From theological perspectives, the dissertation presents the bill from its emergence in the House of Representatives in 2001, to the culmination of the attachment of PLC 122/2006 to PLS 236/2012 in 2013 (a Senate bill that intends to amend the Penal Code). In the first chapter, we have provided conceptual and statistical data on homophobia, listed government programs that seek to eradicate homophobia in Brazil and in the world, and offered a historical overview of PLC 122/2006. In the second chapter, we have elaborated a discursive survey on the position of religious groups that were contrary to PLC 122/2006, namely evangelicals and Roman Catholics. These voices countered PLC 122/2006 with arguments that revolved around a possible restriction of religious freedom if the bill were approved, which created a conflict of rights between the parties that fought for or against it. Next, the study has been illustrated with a brief survey about the official discourse and positions of other religious voices (specifically the Protestant) that have been debating the issue. In the third chapter, we have addressed the question on how this clash relates to the establishment of the Secular State and the construction of human rights, identifying a conflict of rights generated from the complaint that the intervention of religious voices in politics and in the implementation of public policy injures the concept of Secular State. In contrast, for the evangelicals, a group that explicitly stands in defense of a "Christian morality" and a normative family" of a country with a Christian majority, this rejection also injures democracy and the Brazilian Constitution of 1988, which ensures all the citizen religious freedom and expression. As a right granted by the constitution and a mission evoked by the Christian confession of faith that teaches to uphold life in its diversity above any law or dogma, other church voices, Christian public theologies and ecumenical entities are called to contribute to the debate and the struggle for the human rights of LGBT people. After all, we advocate the ongoing dialogue on the urgent need to criminalize homophobia in Brazil assuming that the work is hard, conflicted and even painful, but the silence that oppresses needs to be broken and reported daily by civil society, churches (in their diversity), public theologies and ecumenical bodies, and, not least, by the representatives of people in Congress for a human construction of human rights of LGBT people.

Um Daten am effektivsten zu nutzen, muss die moderne Ökonometrie ihren Werkzeugkasten an Modellen erweitern und neu denken. Das Feld, in dem diese Transformation am besten beobachtet werden kann, ist die kausale Inferenz. Diese Dissertation verfolgt die Absicht Probleme zu untersuchen, Lösungen zu präsentieren und neue Methoden zu entwickeln Machine Learning zu benutzen, um kausale Parameter zu schätzen. Dafür werden in der Dissertation zuerst verschiedene neuartige Methoden, welche als Ziel haben heterogene Treatment Effekte zu messen, eingeordnet. Im zweiten Schritt werden, basierend auf diesen Methoden, Richtlinien für ihre Anwendung in der Praxis aufgestellt. Der Parameter von Interesse ist der „conditional average treatment effect“ (CATE). Es kann gezeigt werden, dass ein Vergleich mehrerer Methoden gegenüber der Verwendung einer einzelnen Methode vorzuziehen ist. Ein spezieller Fokus liegt dabei auf dem Aufteilen und Gewichten der Stichprobe, um den Verlust in Effizienz wettzumachen. Ein unzulängliches Kontrollieren für die Variation durch verschiedene Teilstichproben führt zu großen Unterschieden in der Präzision der geschätzten Parameter. Wird der CATE durch Bilden von Quantilen in Gruppen unterteilt, führt dies zu robusteren Ergebnissen in Bezug auf die Varianz. Diese Dissertation entwickelt und untersucht nicht nur Methoden für die Schätzung der Heterogenität in Treatment Effekten, sondern auch für das Identifizieren von richtigen Störvariablen. Hierzu schlägt diese Dissertation sowohl die „outcome-adaptive random forest“ Methode vor, welche automatisiert Variablen klassifiziert, als auch „supervised randomization“ für eine kosteneffiziente Selektion der Zielgruppe. Einblicke in wichtige Variablen und solche, welche keine Störung verursachen, ist besonders in der Evaluierung von Politikmaßnahmen aber auch im medizinischen Sektor wichtig, insbesondere dann, wenn kein randomisiertes Experiment möglich ist. / To use data effectively, modern econometricians need to expand and rethink their toolbox. One field where such a transformation has already started is causal inference. This thesis aims to explore further issues, provide solutions, and develop new methods on how machine learning can be used to estimate causal parameters. I categorize novel methods to estimate heterogeneous treatment effects and provide a practitioner’s guide for implementation. The parameter of interest is the conditional average treatment effect (CATE). It can be shown that an ensemble of methods is preferable to relying on one method. A special focus, with respect to the CATE, is set on the comparison of such methods and the role of sample splitting and cross-fitting to restore efficiency. Huge differences in the estimated parameter accuracy can occur if the sampling uncertainty is not correctly accounted for. One feature of the CATE is a coarser representation through quantiles. Estimating groups of the CATE leads to more robust estimates with respect to the sampling uncertainty and the resulting high variance. This thesis not only develops and explores methods to estimate treatment effect heterogeneity but also to identify confounding variables as well as observations that should receive treatment. For these two tasks, this thesis proposes the outcome-adaptive random forest for automatic variable selection, as well as supervised randomization for a cost-efficient selection of the target group. Insights into important variables and those that are not true confounders are very helpful for policy evaluation and in the medical sector when randomized control trials are not possible.

Estima-se que 3% da população mundial esteja infectada pelo vírus da hepatite C (HCV). O HCV tem como alvo o tecido hepático e a maioria dos pacientes infectados desenvolvem infecção crônica. Nos últimos anos, estudos in vitro têm demonstrado interações entre o miRNA-122 (miR-122) da célula hospedeira e dois sítios localizados na região 5\' UTR do genoma do vírus da hepatite C (HCV), os quais são essenciais ao processo de replicação viral. O miR-122 é altamente expresso no fígado, onde atua na regulação do metabolismo de lipídios juntamente com outro miRNA, o miRNA-33a (miR-33a), porém, o mecanismo envolvido nesta regulação ainda é pouco conhecido. Sabe-se que a infecção pelo HCV altera a expressão de genes envolvidos na biossíntese e transporte de lipídios, resultando na estimulação do metabolismo de lipídios e criando um ambiente favorável para sua replicação. Neste contexto os objetivos deste trabalho foram avaliar a expressão de miR-33a e miR-122 em indivíduos cronicamente infectados pelo HCV-1 e HCV-3 em amostras obtidas antes do início da terapia. Os miRNAs foram isolados a partir de amostras de sangue periférico e de tecido hepático. A quantificação da expressão relativa de ambos miRNAs foi pela técnica de PCR em tempo real. Os níveis de miR-33a no sangue periférico foram mais elevados do que no tecido hepático em indivíduos infectados pelo HCV-1(p < 0,0001) e HCV-3 (p=0,0025). Observou-se uma correlação inversa entre os níveis de miR-33a no sangue periférico e tecido hepático dos indivíduos infectados pelo HCV-1 (r=-0,281, p=0,039) e correlação positiva para os indivíduos infectados pelo HCV-3 (r=0,9286, p < 0,0001). Correlação inversa entre os níveis hepáticos de miR-33a com o nível sérico de insulina (r=-0,371, p =0,005) nos indivíduos infectados pelo HCV-1 e correlação positiva entre os níveis no sangue periférico com os níveis séricos de GGT (r=0,553, p=0,049) foram observadas. Em relação ao miR-122, de maneira geral o nível hepático foi mais elevado do que o sérico (p < 0,0001). Entretanto, o nível hepático de miR-122 em indivíduos infectados pelo HCV-3 foi maior quando comparado aos infectados pelo HCV-1 (6,22 vezes, p < 0,001). Uma correlação inversa entre os níveis séricos de ApoA-II e os níveis de expressão de miR-122 no sangue (r=-0,330; p=0,014) e tecido hepático (r=-0,311; p=0,020) foi observada nos pacientes infectados pelo HCV-1. Os pacientes infectados pelo HCV- 3 mostraram correlação positiva entre os níveis hepáticos de miR-122 e os níveis de HDL (r=0,412, p=0,036) e insulina (r=0,478, p=0,044). O miR-33a e o miR-122 atuam regulando genes que controlam o metabolismo dos lipídios no fígado. Até o presente momento, não existem relatos que associem a expressão do miR-33a e do miR-122 com o perfil lipídico na infecção pelo HCV. Além disso, o acúmulo de lipídio (esteatose) intensamente descrito na infecção pelo HCV-3 pode sugerir interação diferenciada desse genótipo com os mecanismos envolvidos na regulação do metabolismo lipídico, envolvendo o miR-33a e miR-122 / The prevalence of infection by hepatitis C virus (HCV) is about 3% of the world population. HCV targets the liver tissue and the majority of infected patients develop chronic infection. In recent years, in vitro studies have demonstrated interactions between miRNA-122 (miR-122) the host cell to two places located in the 5\' untranslated region of the HCV genome which are essential for virus replication process. miR-122 is highly expressed in the liver, which has been implicated as a fatty acid metabolism regulator. Another mine has also been described as a key regulator of lipid metabolism, miRNA-33a (miR-33a), however, the mechanisms involved in this regulation are still little known. It is known that HCV infection changes the expression of genes involved in the biosynthesis and transport of lipids, resulting in stimulation of the lipid metabolism and creating a favorable environment for replication of the virus. To our knowledge, there are no reports linking the expression of miR-33a with lipid profile in HCV infection. In this context the objectives of this study were to evaluate the expression of miR-33a and miR-122 in chronically infected individuals with HCV-1 and HCV-3 in samples obtained prior to initiation of therapy. MiRNAs were isolated from peripheral blood samples and liver tissue. The quantification of relative expression of both miRNAs was by PCR in real time. MiR-33a levels in peripheral blood were higher than in liver tissue in patients infected with HCV-1 (p < 0.0001) and HCV-3 (p=0.0025). Levels in the peripheral blood of miR-33a were lower in patients infected with HCV-3 (p=0.0169). There was an inverse correlation between hepatic levels of miR-33a with serum insulin levels (p=0.005) in individuals infected with HCV-1 and a positive correlation between the levels in the peripheral blood serum levels of GGT (p=0.049). Hepatic levels of miR-122 were higher than the levels in the peripheral blood of individuals infected by HCV-1 and HCV-3 (p < 0.0001). Hepatic miR-122 levels were higher in patients infected with HCV-3 than those infected with HCV-1 (6.22 times, p < 0.001). There was a positive correlation between miR-122 levels in the blood and liver tissue of patients infected with HCV-1 (r=0.302, p=0.026). An inverse correlation between serum ApoA-II was observed in these patients the levels of expression of miR-122 in blood (r=-0.330; p =0.014) and liver tissue (r=-0.311; p=0.020). Patients infected with HCV-3 showed a positive correlation between hepatic miR-122 levels to HDL levels (r=0.412, p=0.036) and insulin levels (r=0.478, p=0.044). The miR-33a and miR-122 act by regulating genes that control lipid metabolism in the liver. The different interactions with lipid metabolism exerted by HCV-3 may explain why his relationship with the miR-33a and miR-122 was different when compared with HCV-1