European unions : The Spanish wife and the Scottish widow in Shakespeare and Fletcher's Henry VIII and Ford's Perkin Warbeck

This paper examines Shakespeare’s Henry VIII (1623) and John Ford’s Perkin Warbeck (1634), to assess the extent to which European royal marriage alliances were promoted or interrogated as a means of strengthening diplomatic ties. It pays close attention to the portrayal of the marriages of European women, chiefly the Scottish Katherine Gordon and the Spanish Katherine of Aragon, and their assimilation into the English political sphere. In so doing, it uncovers theatrical suggestions that traditional ideas of European royal marriage – that it adds lustre to the English crown – were tempered with a championing of the relative virtues of Englishwomen. Thus, the two plays engage with a growing sense of English nationalism, as they represent the Scot and the Spaniard as potentially subversive influences. In Henry VIII, Katherine of Aragon, aged and querulous, appeals unsuccessfully to her imagined continental power; in Perkin Warbeck, Katherine Gordon resists the overtures of Henry VII and retains a dangerous autonomy as a widow in an England still uncertain of its own identity

"What think you of this present state?" : Representations of Scotland and Scottish union in Robert Greene's The Scottish History of James the Fourth and John Ford's Perkin Warbeck

Given recently-renewed interest in the political union between Scotland and England, it will be the purpose of this paper to investigate the way in the politically-charged early modern English stage engaged with and represented Scotland and Scots both prior to and following the first iteration of union: that of the countries’ crowns. Consequently, investigation will focus on Robert Greene’s remarkably ahistorical The Scottish History of James the Fourth (c1590) and John Ford’s Perkin Warbeck (c1634). Commonalities in both plays certainly exist – not least the theatrical representation of Scotland’s early-Renaissance king (albeit with wildly differing degrees of historical accuracy). In both plays one finds competing narratives of dominance, division, unity, nationalism, unionism, assimilation, resistance, might and weakness. Yet the plays also offer alternative views of history and British union, with Greene’s largely-fictional, teleological study considering the inevitability of monarchical union and the necessity of English-based dominance (via the collapsing of the historical with the contemporary), and Ford meditating historically on thirty years of that union, positing the imperfections of unified and centralised polity across the British Isles, and raising questions about military might at home versus political might on the European stage. As the early modern theatre’s views of history shifted, so too did views of Scotland. Through these plays, one can identify the nation as a potentially problematical neighbour to be secured by English chastisement and one of several British regions possessed of dangerously independent military might but lacking the political might reserved to the English capita

Slanderisation and censure-ship : when good texts went bad in early modern England

In 1653, the playwright, poet and antiquarian Arthur Wilson’s The History of Great Britain, being the Life and Reign of King James I was published. The text included a reflection on what had become an axiom of political culture in the Stuart age: that poetry and libel had an interdependent relationship. To Wilson, peace begot plenty, and plenty begot ease and wantonness, and ease and wantonness begot poetry, and poetry swelled to that bulk in his time, that it begot strange monstrous satires against the King’s own person, that haunted both court and country. (Wilson 1653: 289-90) Wilson’s jaundiced view of poetry seems fanciful; yet the rise of the verse libel in the Stuart era is well documented. Not only do legal reports of the early modern period recognise the problematic growth of libel, but recent scholarship has made significant inroads in tracing the blossoming of verse libels as a distinct and multifaceted cultural mode, often containing licentious accounts of individuals or political events (Hawarde, Reportes: 143). Indeed, the developing vehicle of the verse libel rapidly became, as Andrew McRae notes, a recognised feature of political and literary culture in the Stuart age (2004a: 1). These often pithy little poems were naturally anathema to the law, not least because they were characterised by their invariably anonymous manuscript circulation. Anonymity itself proved to be an extremely useful means of circumventing legal reprisal, and explosive libels ‘were at one and the same time both written and spoken, simultaneously oral and textual’; and yet the relative success of adopting anonymity as a means of circumvention was not one with which Elizabethan slanderers were routinely armed (Fox 1994: 65). Instead, it emerged gradually, and was met with a flexible legal system that was willing to prosecute anyone who could be found to have knowledge of material deemed seditious or slanderous, whether they were its creators or not

Novel expression and production of Foot-and-mouth disease virus vaccine candidates in Nicotiana benthamiana

Foot-and-mouth disease, also known as FMD, is caused by the aphthovirus Foot-and-mouth disease virus (FMDV), which is a highly contagious disease of cloven-hoofed animals. It is endemic in Africa, parts of South America and southern Asia. In South Africa, the disease is controlled essentially through prophylactic vaccination. Current vaccines on the market are chemically inactivated virus strains. However, these are not considered ideal due to the possibly insufficient inactivation which could fail to render the virus harmless. Research on recombinant vaccines which obviate the need for high biosafety requirements for vaccine preparation has shown that recombinant FMDV virus-like particles (VLPs), devoid of viral genetic material, are an ideal vaccine candidate as they are as immunogenic as the virions themselves when administered to animals. These VLPs are formed by the assembly of the FMDV capsid proteins VP0, VP1 and VP3, which are generated upon the proteolytic cleavage of the capsid precursor protein P1-2A, by the FMDV 3C-protease. The expression platforms used to co-express and produce the component capsid proteins and the protease are usually mammalian, insect or E. coli cells. The use of these expression systems requires extensive bioreactor infrastructure and sterile conditions for vaccine preparation which are costly. In addition, some studies have shown how the co-expression of the 3C-protease can prove to be deleterious when expressed at a high concentration in expression systems. In order to circumvent this, and encourage more efficient production of the capsid proteins and subsequent VLP assembly, researchers have shown that the levels of the 3C-protease can be down-regulated by introducing mutations in the 3C gene or a ribosomal frameshift in the gene sequence which subsequently reduce its deleterious effect. Our laboratory has previously shown that similar FMDV VLPs can be assembled by the expression of FMDV P1-2A (referred to as oP1-2A, in this study), in the absence of the 3C-protease in the plant Nicotiana benthamiana, albeit in low amounts. This platform does not require high biocontainment facilities for the production of recombinant proteins and VLPs and the process is easily scalable. This study centers mainly on optimisation of the FMDV capsid protein expression in N. benthamiana in order to increase VLP yields. I first used codon-optimisation as an approach to improve expression of the capsid proteins and compared expression in the presence and absence of the 3C-protease, using mP1-2A-3C (a new codon-optimised construct), mP1-2A and oP1-2A. Electron microscopy (EM) showed that VLPs resulting from the expression of both mP1-2A-3C and mP1-2A were very low in yield, and irregular in shape and size compared to those produced using oP1-2A. The stability of the plant-produced VLPs was assessed by counting numbers of VLPs, when it was seen that expression of mP1-2A-3C compared to oP1-2A produced an average of 1 VLP per field of view versus 3 VLPs per view, for the same magnification. Furthermore, maturation trials at room temperature was performed on the oP1-2A VLPs, whereby a time-point between 30 to 45 minutes was considered ideal to produce stable VLPs. It is also known that FMDV, unlike the other members of the Picornaviridae family, is acid and heat-labile. The second aim of this study was to promote the stability, and hence encourage the amount of the VLPs produced, by engineering acid and heat-resistant mutants, namely, VP1 N17D, VP2 H93C and VP1 N17D/VP4 S73N using site-directed mutagenesis. A fourth mutant, VP3 A118V which is acid sensitive was used as a control in downstream experiments. The mutants were subjected to a lower than normal pH and a higher than normal temperature. Expression of oP1-2A from the pH and heat assays was assessed to be better than its mutants. The optimum VLP count of 3 VLPs per field of view, was achieved from expression of oP1-2A, after treatment at pH 6.2, compared to 2 VLPs or 1 VLP per field of view for the other mutants tested under all the different conditions. The final aim of this study was to test the immunogenicity of the VLPs from expression of oP1-2A in Balb/C mice. Due to the low yields of VLPs obtained from purification through a continuous gradient, a partial purification method was adopted. Two experimental groups of animals were either vaccinated with P1-2A VLPs or with adjuvanted P1-2A VLPs. A control group was administered with partially-purified plant extract, previously infiltrated with pEAQ-HT. The two experimental groups elicited a marginal increase in humoral immune response at 41 days post vaccination (dpv), which increased significantly at 58 dpv. To my knowledge, this is the first study showing that VLPs produced from expression of FMDV P1-2A only, in tobacco plants, can withstand otherwise degradative acidic and heat conditions. This characteristic has potential for extending the shelf-life of such a candidate vaccine. I also implemented maturation steps to further promote the stability of such VLPs. Finally, the partially purified VLPs showed that they stimulate a significant FMDV P1-2A-specific immune response, particularly in combination with the adjuvant Montanide suggesting that it has potential as a candidate FMDV vaccine

Visualising the Global Structure of Search Landscapes: Genetic Improvement as a Case Study

The search landscape is a common metaphor to describe the structure of computational search spaces. Different landscape metrics can be computed and used to predict search difficulty. Yet, the metaphor falls short in visualisation terms because it is hard to represent complex landscapes, both in terms of size and dimensionality. This paper combines Local Optima Networks, as a compact representation of the global structure of a search space, and dimensionality reduction, using the t-Distributed Stochastic Neighbour Embedding (t-SNE) algorithm, in order to both bring the metaphor to life and convey new insight into the search process. As a case study, two benchmark programs, under a Genetic Improvement bug-fixing scenario, are analysed and visualised using the proposed method. Local Optima Networks for both iterated local search and a hybrid genetic algorithm, across different neighbourhoods, are compared, highlighting the differences in how the landscape is explored

A Comparison of Operator Utility Measures for On-Line Operator Selection in Local Search

This paper investigates the adaptive selection of operators in the context of Local Search. The utility of each operator is computed from the solution quality and distance of the candidate solution from the search trajectory. A number of utility measures based on the Pareto dominance relationship and the relative distances between the operators are proposed and evaluated on QAP instances using an implied or static target balance between exploitation and exploration. A refined algorithm with an adaptive target balance is then examined

How Perturbation Strength Shapes the Global Structure of TSP Fitness Landscapes

Local optima networks are a valuable tool used to analyse and visualise the global structure of combinatorial search spaces; in particular, the existence and distribution of multiple funnels in the landscape. We extract and analyse the networks induced by Chained-LK, a powerful iterated local search for the TSP, on a large set of randomly generated (Uniform and Clustered) instances. Results indicate that increasing the perturbation strength employed by Chained-LK modifies the landscape's global structure, with the effect being markedly different for the two classes of instances. Our quantitative analysis shows that several funnel metrics have stronger correlations with Chained-LK success rate than the number of local optima, indicating that global structure clearly impacts search performance