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Fanconi Anaemia (FA) is an inherited autosomal-recessive disorder that can lead to abnormal development, bone-marrow failure, and an increased vulnerability to carcinogenesis. Cells derived from FA patients are unusually sensitive to DNA crosslinking agents and it is now known that FA cells lack one of twenty-two known different FA complementation group (FANC) proteins involved in the repair of DNA interstrand crosslinks (ICLs). More recently, two DNA glycosylases (endonuclease VIII - like 1 & 3 [NEIL1 & NEIL3]) that excise oxidised bases from DNA, thus initiating base excision repair, have been found to participate in the resolution of psoralen induced ICLs. Therefore, this project set out to determine whether the modulation of NEIL1 or NEIL3 expression in cell lines lacking FANCD2, of the monoubiquitinated DNA-heterodimer protein complex FANCD2/FANCI, could significantly affect their sensitivity to ICL-inducing agents, such as mitomycin C and cisplatin. An FA original and two FA-generated cancerous cell model systems representing FA and equivalent wildtype cells were then transfected with plasmids expressing FLAG-tagged NEIL1 or NEIL3 and challenged with the crosslinking agents or the oxidising agent tert-butyl hydroperoxide, and any differences in sensitivity to these agents determined. Similarly, siRNAs were designed against NEIL3 and the E3-ubiquitin ligase tripartite motif-containing protein 26 (TRIM26), that has been shown to ubiquinate both NEIL1 and NEIL3 for proteasomal degradation and similar cell growth analyses performed using the MTT assay. Recombinant NEIL1 expression was confirmed by western blotting but had little effect on resistance to crosslinking and oxidising agents. Overexpression of NEIL3, either a truncated version lacking the C-terminal GRF domains (NEIL31506), or the full-length protein, proved more difficult to confirm by western blotting, possibly due to antibody specificities to recombinant and non-recombinant NEIL3. However, MTT assays generally indicated a small increase in resistance irrespective of FA background, suggesting that NEIL3 overexpression was achieved in these cells. Unexpectedly, compared to literature and Elder laboratory results, knockdown of NEIL3 showed increased resistance against ICL and oxidative agents independent of FA phenotype, while knockdown of TRIM26 showed no clear effect on the sensitivity of the FA cell line to the genotoxic agents tested. In conclusion, the results indicate that the overexpression of NEIL1 or NEIL3 could not compensate for the loss of the FA pathway of ICL repair and had little reproducible effect on the resistance of these cell lines to crosslinking and oxidizing agents.

The primary purpose and content of this thesis consists of 'editiones principes' of twenty-five Jewish Babylonian Aramaic magic bowl texts from the collection of the Vorderasiatisches Museum (Berlin), including, for each text: transcription; English translation; philological notes to justify any difficult aspects of the given transcription and translation; sketches of any artwork; and commentary discussing the notable features of each bowl, in terms of both the textual content and the extratextual. The editions are followed by glossaries listing and parsing all the word forms found in the text. As well as an introductory literature review describing the progress of the field of the study of magic bowls from its beginnings to the present day, the thesis also includes a chapter discussing how the magic of the bowls works. Herein it is argued that the magic is designed to influence the actions of supernatural beings, most commonly demons, by tapping into the power of God to whom they, as all things, are already subject. Commonly this is done by appealing to precedents of what God has previously done or said or his known character traits, under the premise that he can be relied upon to be consistent with himself. Further techniques at the disposal of bowl scribes were the use of Biblical quotations; powerful names, including everything from names of God and specific angels to strings of apparent gibberish designed to tap into the creative power of the alphabet itself; ‘seals’ on the text, articulated in writing; conventions borrowed from legal practice; and various non-textual drawings: all these were designed to establish and augment the authority according to which the targeted supernatural beings were being commanded. This approach to magic may be characterised as both distinctly Mesopotamian and profoundly monotheistic.

The deformation front of an orogen is defined as a zone or surface separating regions of different deformation styles or strain intensities. The large-scale development and migration of deformation front structures can be described and understood using critical wedge theory. At the foreland of a fold-thrust belt, the deformation front can be marked by a number of structures - an imbricate fan or frontal monocline, a ramp anticline or tip-line fold pair or a passive-roof duplex (triangle zone). This study considers two fold-thrust belts, the Zagros Simply Folded Belt, Iran and the Sawtooth Range, Montana. Both regions are developed in a passive margin carbonate sequence, overlain by a clastic succession. Remote sensing and field observations, combined with the interpretation of seismic data, are used to define the variation in fold structures within the belts and at the deformation fronts, and to constrain the migration of the deformation fronts. Palaeo-deformation fronts in the Zagros Simply Folded Belt are dominantly characterised by ramp anticlines, but the recent deformation front is marked in part by a reactivated fault structure above a salt diapir. The deformation front along the majority of the Sawtooth Range is a classic imbricate fan sequence, however, a triangle zone is observed in at least one segment of the belt. The development of each of these classes of structures is strongly influenced by the nature of inherited basement structures as well as by the mechanical stratigraphy of the sediment pile and the detachment type. The field and remote sensing components of the study are complemented by an analogue modelling study which is designed to investigate one of these factors, namely the effect of varying the geometry of an upper detachment horizon within the sediment pile. Results imply that the geometry of the upper detachment exerts a first order control on the development of deformation front structures. Triangle zones are found to develop where the upper detachment is narrow with respect to the width of the deformed zone. Conversely, foreland-verging structures develop where this upper detachment is wide with respect to the deformed zone.. When model results are compared to the field areas described above, this effect is shown to be a valid explanation for the structures developed. These results imply thatforeland triangle zones may be more common than currently documented, which has additional implications for hydrocarbon exploration and production. In addition, this thesis demonstrates the value of remote sensing methods for both hydrocarbon and mineral exploration.

Pattern recognition receptors (PRRs) sense unique and conserved structures common to many types of microorganisms and activate pro-inflammatory signaling cascades that are important for mounting effective immune responses, but aberrant regulation of PRR signaling can lead to autoimmunity. Polymorphisms in NOD2, an intracellular PRR, are associated with increased risk of developing the inflammatory bowel disorder Crohn’s disease. To gain insight into how NOD2 signaling is regulated, we performed a genome-wide siRNA screen in which NOD2-induced NF-κB activation was assessed using a luciferase reporter system. Using this strategy, we identified and validated hundreds of novel regulators of the NOD2 and NF-κB signaling pathway, some of which were previously implicated in Crohn’s disease susceptibility. By comparing our results with publicly available protein-protein interaction databases, we were able to visualize networks of genes, including the linear ubiquitin chain assembly complex, that are important for mediating NOD2-dependent responses. Classically, PRRs are thought to function in differentiated cells to orchestrate inflammation, but recent evidence suggests that hematopoietic stem cells (HSCs), which maintain production of all blood cells throughout life, express PRRs and could be important for supporting immune responses during infection though the mechanisms underlying this function are unclear. We found that systemic infection of mice with the gram negative bacterium Escherichia coli resulted in moderate reduction of HSC activity in bone marrow and vastly expanded HSC activity in spleen, indicative of extramedullary hematopoiesis. Expansion of splenic HSCs was reduced in mice deficient for the PRR TLR4 or the adaptor protein RIPK2, which is required for NOD1 and NOD2 signaling, implicating PRRs in the regulation of HSCs during infection. PRR signaling in radio-resistant cells was important for promoting progenitor expansion in spleen, suggesting an indirect mechanism. We found that expansion of splenic HSCs was dependent on dual activation of NOD1 and TLR4 and production of granulocyte-colony stimulating factor. Altogether, these results provide a global view of the mechanisms that regulate PRR signaling and provide insight into the pathways that facilitate extramedullary hematopoiesis in the context of infection.

Current treatment for type I diabetes includes delivery of insulin via injection or pump, which is highly invasive and expensive. The production of chloroplast-derived proinsulin should reduce cost and facilitate oral delivery. Therefore, tobacco and lettuce chloroplasts were transformed with the cholera toxin B subunit fused with human proinsulin (A, B, and C peptides) containing three furin cleavage sites (CTB-PFx3). Transplastomic lines were confirmed for site-specific integration of transgene and homoplasmy. Old tobacco leaves accumulated proinsulin up to 47% of total leaf protein (TLP). Old lettuce leaves accumulated proinsulin up to 53% TLP. Accumulation was so stable that up to ~40% proinsulin in TLP was observed even in senescent and dried lettuce leaves, facilitating their processing and storage in the field. Based on the yield of only monomers and dimers of proinsulin (3 mg/g leaf, a significant underestimation), with a 50% loss of protein during the purification process, one acre of tobacco could yield up to 20 million daily doses of insulin per year. Proinsulin from tobacco leaves was purified up to 98% using metal affinity chromatography without any His-tag. Furin protease cleaved insulin peptides in vitro. Oral delivery of unprocessed proinsulin bioencapsulated in plant cells or injectable delivery into mice showed reduction in blood glucose levels similar to processed commercial insulin. C-peptide should aid in longterm treatment of diabetic complications including stimulation of nerve and renal functions. Hyper-expression of functional proinsulin and exceptional stability in dehydrated leaves offer a low cost platform for oral and injectable delivery of cleavable proinsulin.