Replication enhancer elements within the open reading frame of tick-borne encephalitis virus and their evolution within the Flavivirus genus

We provide experimental evidence of a replication enhancer element (REE) within the capsid gene of tick-borne encephalitis virus (TBEV, genus Flavivirus). Thermodynamic and phylogenetic analyses predicted that the REE folds as a long stable stem–loop (designated SL6), conserved among all tick-borne flaviviruses (TBFV). Homologous sequences and potential base pairing were found in the corresponding regions of mosquito-borne flaviviruses, but not in more genetically distant flaviviruses. To investigate the role of SL6, nucleotide substitutions were introduced which changed a conserved hexanucleotide motif, the conformation of the terminal loop and the base-paired dsRNA stacking. Substitutions were made within a TBEV reverse genetic system and recovered mutants were compared for plaque morphology, single-step replication kinetics and cytopathic effect. The greatest phenotypic changes were observed in mutants with a destabilized stem. Point mutations in the conserved hexanucleotide motif of the terminal loop caused moderate virus attenuation. However, all mutants eventually reached the titre of wild-type virus late post-infection. Thus, although not essential for growth in tissue culture, the SL6 REE acts to up-regulate virus replication. We hypothesize that this modulatory role may be important for TBEV survival in nature, where the virus circulates by non-viraemic transmission between infected and non-infected ticks, during co-feeding on local rodents.

Access to the countryside of England and Wales: popular leisure; commodified culture

Risk and uncertainty are, to say the least, poorly considered by most individuals involved in real estate analysis - in both development and investment appraisal. Surveyors continue to express 'uncertainty' about the value (risk) of using relatively objective methods of analysis to account for these factors. These methods attempt to identify the risk elements more explicitly. Conventionally this is done by deriving probability distributions for the uncontrolled variables in the system. A suggested 'new' way of "being able to express our uncertainty or slight vagueness about some of the qualitative judgements and not From its modern origins, associated with the urbanising effect of industrialisation, walking has remained a popular form of outdoor recreation. It has, furthermore, remained an important site of class struggle, with the 'landless' seeking to establish their moral 'citizen' right to roam over open country in contradistinction to the 'landed', who have successfully limited this right to legally-defined public rights of way. In the face of declining farm incomes, however, farmers and landowners have, apparently, modified their attitudes towards public access, but only in return for compensation and management payments under grant schemes such as Countryside Stewardship and the Countryside Premium Scheme. With the Ministry of Agriculture, Fisheries and Food now seeking to extend paid access arrangements to other grant schemes, as part of its response to the European Union's Agri-Environment Regulations, access 'rights' are assuming an increasingly commodified form, thereby questioning, if not undermining, the former citizen claims. For rather than being a benefit of citizenship, the existence of limited, often poorly maintained and inadequately signposted, public rights of way has tied inextricably the extension of legally-enforceable access to the needs of the landowners and farmers. At a time of falling prosperity in agriculture, therefore, they have now exercised their discretion by annexing the populism of consumer culture to reproduce the bourgeois liberal values of the market as a principal determinant of the extension of citizen rights of access to the countryside.

Expectation versus provision in the management and use of urban open space

This paper discusses the implications of the shifting cultural significance of public open space in urban areas. In particular, it focuses on the increasing dysfunction between people's expectations of that space and its actual provision and management. In doing so, the paper applies Lefebvre's ideas of spatiality to the evident paradigm shift from 'public' to 'private' culture, with its associated commodification of previously public space. While developing the construct of paradigm shift, the paper recognises that the former political notions inherent in the provision of public space remain in evidence. So whereas public parks were formerly seen as spaces of confrontation between the 'rationality' of public order as the 'irrationality' of individual leisure pursuits, they are now increasingly seen, particularly 'out of hours', as the domain of the dispossessed, to be defined and policed as 'dangerous'. Where once people were welcomed into public open spaces as a means of 'educating' them in good, acceptable, leisure practices, therefore, they are now increasingly excluded, but for the same ostensible reasons. Building on survey work undertaken in Reading, Berkshire, the paper illustrates how communities can become separated from 'their' space, leaving them with the overriding impression that they have been 'short-changed' in terms of both the provision and the management of urban open space. Rather than the intimacy of local space for local people, therefore, the paper argues that parks have become externalised places, increasingly responding to commercial definitions of culture and what is 'public'. Central urban open spaces are therefore increasingly becoming sites of stratification, signification of a consumer-constructed citizenship and valorisation of public life as a legitimate element of the market surface of town and city centres.

Re-fitting Etton: space, time and material culture within a causewayed enclosure in Cambridgeshire

This paper considers the dynamics of deposition around and across the causewayed enclosure at Etton, Cambridgeshire. As a result of detailed re-analysis (particularly refitting) of the pottery and flint assemblages from the site, it proved possible to shed new light both on the temporality of occupation and the character of deposition there. Certain aspects of our work challenge previous interpretations of the site, and of causewayed enclosures in general; but, just as importantly, others confirm materially what has previously been suggested. The quantities of material deposited at Etton reveal that the enclosure was occupied only very intermittently and certainly less regularly than other contemporary sites in the region. The spatial distribution of material suggests that the enclosure ditch lay open for the entirety of the monument's life, but that acts of deposition generally focused on a specific part of the monument at any one time. As well as enhancing our knowledge of one particular causewayed enclosure, it is hoped that this paper – in combination with our earlier analysis of the pit site at Kilverstone – makes clear the potential that detailed material analysis has to offer in relation to our understanding of the temporality of occupation on prehistoric sites in general.