Activity-dependent and synapse-specific translation of mRNAs is required for long-term changes in synaptic strength (or efficacy). However, many of the components mediating repression, transport and activation of mRNAs are unknown. Translational control in neurons is a highly conserved process and mediated by a ribonuclear particle (RNP). This study shows that RNPs in Drosophila neurons are similar not only to mammalian neuronal RNA granules but also to yeast P-bodies, cytoplasmic foci involved in translational repression and RNA decay. The evolutionarily conserved proteins Me31b and Trailer Hitch localize to RNA granules. Me31b and Trailer Hitch are required for normal dendritic growth. Mutations in Me31b and Trailer Hitch suppress phenotypes resulting from overexpression of Fragile X Mental Retardation protein, suggesting that both proteins may act as translational repressors. In addition, this study reports the identification of novel translational repressors in neurons. Using the overexpression phenotype of Fragile X Mental Retardation protein in a candidate-based genetic screen, I identified dominant suppressor mutations in five genes, including Doubletime/Discs Overgrown, Orb2/CPEB, PolyA Binding Protein, Rm62/Dmp68 and SmD3. Like Me31b and Trailer Hitch, all five proteins localize to neuronal RNPs. Overexpression of each proteins affects dendritic branching of sensory neurons in Drosophila. Identification and further characterization of these novel RNP granule components and dFMR1-interacting proteins may provide further insights into the mechanisms controlling translational in dendrites.

The effectiveness of radiotherapy treatment could be significantly improved if tumour cells could be rendered more sensitive to ionising radiation without altering the sensitivity of normal tissues. However, many of the key mechanisms that determine intrinsic tumour radiosensitivity are largely unknown. This thesis is concerned with the identification of novel determinants of tumour radiosensitivity. A siRNA screen of 200 genes involved in DNA damage repair was conducted using γH2AX foci post-irradiation as a marker of cell damage. This screen identified POLQ as a potential tumour-specific contributor to radioresistance. Subsequent investigations demonstrated that POLQ knockdown resulted in radiosensitisation of a panel of tumour cell lines, whilst having little or no effect on normal tissue cell lines. It was subsequently shown that POLQ depletion rendered tumour cells significantly more sensitive to several classes of cytotoxic agents. Following exposure to etoposide, it was found that tumour cells depleted of POLQ had reduced RAD51 foci formation, suggesting that POLQ is involved in homologous recombination. A homologous recombination assay was used to confirm that POLQ depletion does indeed result in reduced homologous recombination efficiency. These findings led to the investigation of the clinical significance of tumour overexpression of POLQ. The clinical outcomes of patients with early breast cancer were correlated with tumour expression levels of POLQ. It was found that POLQ overexpression was correlated with ER negative disease and high tumour grade, both of which are associated with poor clinical outcomes. POLQ overexpression was associated with extremely poor relapse free survival rates, independently of any other clinical or pathological feature. The mechanism that causes this adverse outcome may in part arise from resistance to adjuvant chemotherapy and radiotherapy treatment. These findings, combined with the limited normal tissue expression of POLQ, make it an appealing target for possible clinical exploitation.

This thesis chronicles the progression toward and creation of my thesis show, Limelight & Indigestion, as well as the cultural, technological and artistic influences and discussions that underpin the works therein. The show is an exploration of celebrity, mass media, and the nature of the desire for fame. I hope to situate my work through an investigation of topics such as Hollywood and the use of green screens and associated technology, our physical and ideological connection to cinema, the absurd in cinema history, and the complex nature of media digestion.

Answering the call issued by John Underkoffler in 2010 about the future of UI, I have imagined the Immersive Media Library (IML) as an annex of the main VCU library, offering a concentration of visually immersive spaces to compliment the space the university is already building in the renovated Cabell Library. The design is new in that the emphasis is placed on the collaboration between librarians and visitors in creating new work. Focusing on the interpersonal might be unexpected from program with such an emphasis on new technology - but I see it as vital part of the new computing paradigm.

Melanocytes are pigment-producing cells that colour our hair, skin and eyes. Melanocytes are evolutionary conserved in vertebrates, and in addition to contributing to pigmentation and pattern formation, can contribute to background adaptation (zebrafish) and protection against harmful UV irradiation (humans). Many of the processes involved in melanocyte development – such as migration, proliferation and differentiation - are misregulated in melanoma. Here, I use chemical biology in zebrafish to identify targetable pathways in melanocyte development and regeneration, with a view to how these processes may be misregulated in melanoma and other pigmentation syndromes. We first wanted to address the potential for small molecules to regulate multiple stages of melanocyte development and differentiation. In Chapter 3, I describe my work involved in a small molecule screen for clinically active compounds that alter melanocyte biology (Colanesi et al., 2012). In this work we have identified small-molecules that affect melanocyte migration, differentiation, survival, morphology and number. This is important as it highlights new pathways essential for normal melanocyte development and consequently provides further tools in which to study melanocytes. Identifying the target of small molecules in vivo is a challenge in chemical biology. In Chapter 4, I describe my contributions to understanding how 5-nitrofuran compounds act in zebrafish (Zhou et al., 2012). My work has contributed to understanding the activity of 5-nitrofurans is dependent upon its nitrofuran ring structure. I have also helped confirm a conserved interaction between 5-nitrofurans and ALDH2, which may contribute to the off-target effects observed in the clinic. These results are important as they aid further understand of the 5-nitrofuran class of drugs and give evidence to support combination therapy of 5-nitrofurans with ALDH2 inhibitors as a way to overcome clinical side effects. Additionally I show that NFN1 treatment limits ensuing melanocyte regeneration thereby suggesting a role at the Melanocyte Stem Cell (MSC), which provides me with a key tool to study melanocyte regeneration in zebrafish. How tissue specific cell numbers are specified and maintained is a key question in developmental biology. In Chapter 5, I describe the identification of the MITF gene in the maintenance of cell cycle arrest in differentiated melanocytes (Taylor et al., 2011). We show that the human melanoma mutation MITF4TΔ2B promotes melanocyte division, thereby suggesting a role for melanocyte division in the pathogenesis of melanoma. This work is valuable because it highlights Mitf as a molecular rheostat that controls melanocyte proliferation and differentiation in living vertebrates, and helps us to understand the role of MITF in melanoma progression. Little is known about the pathways that control melanocyte stem cells in animals. To identify new melanocyte stem cell pathways, I used NFN1 as the basis for a small molecule screen for enhancers of melanocyte regeneration (Chapter 6). I find that chemical inhibition of Phosphatase of Regenerating Liver-3 (Prl-3) in zebrafish can enhance melanocyte regeneration. Importantly, I have found that there are an increased number of melanocyte progenitor cells in PRL3-inhibitor treated zebrafish. I propose that PRL-3 may control progenitor cell number in melanocyte regeneration. This is significant because it identifies PRL-3 as a novel molecular target controlling melanocyte progenitor cells, and identifies a new chemical tool with which to study melanocyte differentiation from a progenitor population. In the final chapter, I discuss how this work relates to the larger field of melanocyte developmental biology, and the new insight it provides into the fundamental processes of how organisms control cell number and pattern formation. In addition, I discuss how this work may have implications for understanding and treating melanocyte diseases, such as vitiligo (loss of melanocytes) and melanoma (cancer of the melanocyte).

Die Verkleinerung der modernen Temperatursensoren ermöglicht die Volumenreduzierung der bisher angewandten Stevenson-Wetterhütten. Insbesondere bei automatischen Stationen finden wir wesentlich kleinere Strahlungsschutzeinrichtungen. Langjährige Beobachtungsreihen sind nicht mehr vergleichbar und müssen angepaßt werden. Es werden Einzelwertabweichungen für Temperaturen von mehr als 1 K genannt. Zur Lösung dieses Problems sind Vergleichsuntersuchungen und Anpassungsrechnungen erforderlich. / The minimisation of modern temperature sensors allows to reduce the volume ofthe up to now used Stevenson screens. Especially for automatic stations we found essential smaller radiationshields. Temperature observations of many years arc incomperable and had to be adapted. Errors of more as 1 K for single temperatures are mentioned. For the solution of this problem comparisons and adaptations are necessary.

Thesis advisor: Marc A.T. Muskavitch / The Notch pathway is an evolutionarily conserved mechanism of intercellular signaling that plays a central role in the development of metazoans. Here I summarize two genetic screens that utilize a rough eye phenotype created by Delta overexpression in the Drosophila eye to identify modulators of Notch pathway signaling activity. Among the many "hits" obtained from both screens, I have mapped to the Not1 gene a single complementation group that exhibits strong genetic interactions with Notch pathway mutants. NOT1 is a component of the CCR4-NOT complex, a global regulator of gene expression that exerts its effects through a variety of mechanisms, including mRNA deadenylation and direct transcriptional repression. I have conducted a series of genetic and molecular experiments in an effort to obtain more insight into the relationship between the CCR4-NOT complex and the Notch pathway. Both Not1 EMS mutations and RNAi-mediated knockdown of NOT1 expression produce phenotypes that mimic those of Notch loss-of-function pathway mutants. Knockdown of NOT1 in the developing bristle organ disrupts Notch-mediated inhibition of neuronal specification, resulting in supernumerary neurons and aberrant sheath cell specification. Knockdown of NOT1 within the developing wing margin disrupts expression of the Notch target genes Cut and Wingless, as well as the Notch ligand Delta. Phenotypic rescue experiments imply that Not1 functions downstream of Notch signal activation and acts directly on Notch target gene expression. These results suggest that NOT1 is required for Notch signal transmission in certain developmental contexts and implicate the CCR4-NOT complex as a positive regulator of the Notch pathway. / Thesis (PhD) — Boston College, 2009. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Biology.

Thesis advisor: Charles S. Hoffman / The biological roles of the most recently discovered mammalian cyclic nucleotide phosphodiesterase (PDE) family, PDE11, are poorly understood, in part due to the lack of selective inhibitors. To address this need for such compounds I completed a ~200,000 compound high throughput screen (HTS) for PDE11 inhibitors using a yeast-based growth assay. Further characterization of lead candidates using both growth-based assays in the fission yeast Schizosaccharomyces pombe and in vitro enzyme assays identified four potent and selective PDE11 inhibitors. I examined the effect of these compounds on human adrenocortical cells, where PDE11 is believed to regulate cortisol levels. One compound, along with two structural analogs, elevates cAMP levels and cortisol production through PDE11 inhibition, thus phenocopying the behavior of adrenocortical tumors associated with Cushing syndrome. These compounds can be used as research tools to study the biological function of PDE11, and can also serve as leads to develop therapeutic compounds for the treatment of adrenal insufficiencies. This study further validates the yeast-based HTS platform as a powerful tool for the discovery of potent, selective and biologically-active PDE inhibitors. / Thesis (PhD) — Boston College, 2012. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Biology.

Thesis advisor: Clare O'Connor / Protein isoaspartyl methyltransferase (PIMT), an ancient enzyme with putative homologs in virtually all eukaryotes studied to date, is a protein repair enzyme that initiates restoration of age-damaged isoaspartyl residues back into normal amino acids. I studied the functional importance of PIMT in Schizosaccharomyces pombe, whose PIMT-encoding homolog is designated pcm2. Past studies of PIMT in other organisms had suggested that Pcm2p function could be important for viability under stress conditions. In the first part of the project, I compared the viability of a yeast strain lacking pcm2 under a variety of stress conditions, including stationary phase, heat shock, high-salt, high-sugar, high-pH, and high temperature. Based on studies of PIMT in other organisms, I expected to see reduced viability of the knockout strain during stress conditions, but no difference in cell growth or viability was observed. I also attempted to use Real-Time PCR to compare pcm2 transcription in yeast under the same stress conditions. I have not yet collected any usable data from the Real-Time PCR runs, but it has been previously shown that pcm2 transcription increases during stationary phase, heat shock, and osmotic shock. This increase in pcm2 transcription during stress with no phenotype associated with loss of pcm2 under stress conditions leads to the hypothesis that there may be a second gene whose product can compensate for loss of PIMT function. To locate and identify this gene, a synthetic lethal screen is being utilized. Currently, five potential synthetic lethal mutants have been isolated. / Submitted to: Boston College. College of Arts and Sciences. / Discipline: College Honors Program. / Discipline: Biology.

Peneiras vibratórias são equipamentos amplamente utilizados em circuitos de cominuição nas mais variadas funções, desde classificação primária, processando blocos de algumas toneladas até em um escalpe de finos para um britador, até mesmo um processo de desaguamento, no qual as telas atuam como filtros, processando polpas que contêm partículas muito finas. Sua utilização tem crescido consideravelmente nos últimos anos, pois quando as peneiras vibratórias são corretamente aplicadas, possibilitam aumentar a eficiência de uma usina a custos razoavelmente baixos. Embora as funções do peneiramento sejam de fácil entendimento, o processo de seleção e dimensionamento dos equipamentos é complexo e sujeito a erros de difícil solução. Quando o minério tem baixa umidade, caso da maioria das aplicações, os dimensionamentos feitos pelos métodos tradicionais apresentam boa correlação com os resultados obtidos nas peneiras industriais. Entretanto, quando a umidade é alta, os cálculos podem apresentar grandes desvios frente ao desempenho real ou simplesmente não funcionar. Este problema é bastante comum em regiões tropicais, como o Norte do Brasil, local com alta incidência de chuvas, onde predominam minérios alterados. O objetivo deste trabalho foi desenvolver um método de dimensionamento de peneira, através de ensaios em laboratório com pequena quantidade de amostras, que permite ao usuário estimar o desempenho de uma peneira industrial no processamento de minérios difíceis com melhor precisão, comparado aos métodos convencionais. As conclusões demonstram a viabilidade técnica do método proposto e seus limites de aplicação. / The vibrating screens are widely used in the comminution circuits for different applications. They can be used for fines removal before a crusher or as a dewatering equipment, where the screening media act as a filter. The screen utilization has been extensively growing in the last years since its accurate operation allows plant efficiency improvements at a minor cost. Although the screening process has an easy understanding, the scale-up and sizing methods can be complex and subject to errors that are difficult to solve. When an ore has low natural moisture, representing most applications, the screen size estimates done by traditional methods normally shows proper correlation compared to industrial screens. However, when the moisture content is high, calculations may present significant deviations when compared to the real operation. This kind of problem is characteristic for tropical regions such as the North of Brazil, where weathered rocks are predominated. The objective of this work was to develop a screen sizing method which allows the user to do an industrial screen performance estimation for difficult high natural moisture ores, based on laboratory test work with reduced quantity of samples, and to provide better accuracy when compared to the results coming from traditional sizing methods. The conclusions demonstrate the technical viability of the method and its application limits.