Rational Attenuation of a Morbillivirus by Modulating the Activity of the RNA-Dependent RNA Polymerase

Negative-strand RNA viruses encode a single RNA-dependent RNA polymerase (RdRp) which transcribes and replicates the genome. The open reading frame encoding the RdRp from a virulent wild-type strain of rinderpest virus (RPV) was inserted into an expression plasmid. Sequences encoding enhanced green fluorescent protein (EGFP) were inserted into a variable hinge of the RdRp. The resulting polymerase was autofluorescent, and its activity in the replication/transcription of a synthetic minigenome was reduced. We investigated the potential of using this approach to rationally attenuate a virus by inserting the DNA sequences encoding the modified RdRp into a full-length anti-genome plasmid from which a virulent virus (rRPV(KO)) can be rescued. A recombinant virus, rRPV(KO)L-RRegfpR, which grew at an indistinguishable rate and to an identical titer as rRPV(KO) in vitro, was rescued. Fluorescently tagged polymerase was visible in large cytoplasmic inclusions and beneath the cell membrane. Subcutaneous injection of 10(4) TCID(50) of the rRPV(KO) parental recombinant virus into cattle leads to severe disease symptoms (leukopenia/diarrhea and pyrexia) and death by 9 days postinfection. Animals infected with rRPV(KO)L-RRegfpR exhibited transient leukopenia and mild pyrexia, and the only noticeable clinical signs were moderate reddening of one eye and a slight ocular-nasal discharge. Viruses that expressed the modified polymerase were isolated from peripheral blood lymphocytes and eye swabs. This demonstrates that a virulent morbillivirus can be attenuated in a single step solely by modulating RdRp activity and that there is not necessarily a correlation between virus growth in vitro and in vivo.

Rational design of a dual-mode optical and chemical prodrug

Purpose. The purpose of this study is to demonstrate the rational design and behaviour of the first dual mode optical and chemical prodrug, exemplified by an acetyl salicylic acid-based system. Methods. A cyclic 1,4-benzodioxinone prodrug was synthesised by reaction of 3,5-dimethoxybenzoin and acetyl salicoyl chloride with pyridine. After purification by column chromatography and recrystallization, characterization was achieved using infrared and NMR spectroscopies, mass spectrometry, elemental analysis and single crystal X-ray diffraction. Light-triggered drug liberation was characterised via UV-visible spectroscopy following low-power 365 nm irradiation for controlled times. Chemical drug liberation was characterised via UV-visible spectroscopy in pH 5.5 solution. Results. The synthetic method yielded pure prodrug, with full supporting characterisation. Light-triggered drug liberation proceeded at a rate of 8.30 10j2 sj1, while chemical, hydrolytic liberation proceeded independently at 1.89 10j3 sj1. The photochemical and hydrolytic reactions were both quantitative. Conclusions. This study demonstrates the first rational dual-mode optical and chemical prodrug, using acetyl salicylic acid as a model, acting as a paradigm for future dual-mode systems. Photochemical drug liberation proceeds 44 times faster than chemical liberation, suggesting potential use in drug-eluting medical devices where an additional burst of drug is required at the onset of infection.

Greek-Turkish Dilemmas and the Cyprus-EU Accession Process

This article compares the processes of foreign policymaking in Greece and Turkey in order to examine why the incentives and pressures of the enlargement process have failed until now to initiate a settlement in the Cyprus bicommunal negotiations. While most studies on the Cyprus problem have focused on the two communities of the island, little at-tention has been paid to the policies of the two â??motherlandsâ??, namely Greece and Turkey. Yet their leverage on the two Cypriot communities and their conflicting expectations with regard to an enlarged Europe in the Eastern Mediterranean constitute a complex security puzzle. The Republic of Cyprus stands as a champion candidate member for the next enlargement, amid fears of Turkish reprisals and hopes for a po-litical settlement on the island. With the benefits of settlement overwhelming the benefits of any other alternative, it is paradoxical that the parties seem to be about to fail to reach a last-minute, mutually beneficial compromise. I try to resolve this paradox by supplementing rational choice theory with cognitivist theories of international relations. While rational choice predicts a direct relationship between external environment and foreign policy shifts, the case of Cyprus suggests that this relationship is actually indirect. Without understanding how the external environment is framed in the domestic political discourse of Greece and Turkey, it is impossible to demonstrate how outside pressure and incentives affect foreign policy shifts.

Is the General Public Licence a Rational Choice?

Open source projects are networks of developers, distributors and end-users of non-proprietary created knowledge goods. It has been argued that this form of organization has some advantages over the firm or market coordination. I show that for sufficiently convex and modular projects proprietary licences are not able to sustain sequential knowledge production which, however, can be carried out if the project is run on the open source basis.

Experiences and expectations: the real reason nobody likes stats

Within undergraduate psychology courses, students often have significant levels of anxiety and negative attitudes toward the statistical element. This has been attributed to poor interaction with teachers, fears about mathematical abilities, and simply being unaware of that portion of the course or its relevance to psychology. To address this, 196 undergraduate psychology students completed a survey on statistics anxiety and attitudes. Additionally, 27 different students in similar situations took part in focus group to share their experiences of introductory statistics courses. Survey results showed that fewer than half were aware of the statistics portion of their course and that the expectation was a key factor in their experiences. Qualitative feedback from the focus groups revealed much about how the teaching may or may not improve attitudes nor decrease anxiety. Findings support various broad strategies (i.e. increase awareness of statistics in psychology and confidence in success in the course) as opposed to skill-specific (better ways of teaching probability or using games to increase participation, for example) classroom interventions to improve statistics education.

Beyond Compliance – Below Expectations? Cross-Border CSR, Development and the UN Global Compact.

In this paper, the results of an empirical analysis of a set of 416 descriptive case studies published by corporate members of the UN Global Compact are presented. Although these cases cannot be viewed as representative of the Compact itself or of corporate social responsibility (CSR) and development in general, they can illustrate which kinds of projects are deemed appropriate as best practice examples among Compact members, and therefore indicate the direction, in which predominantly voluntary and business-led CSR might at best be evolving. To help contextualize the analysis, the paper starts with a brief overview of recent academic work on the strengths and limitations of CSR in the light of international development, followed by the empirical analysis of Compact case studies. The results raise doubts regarding the general suitability of contemporary CSR initiatives to tackle some of the most pressing developmental challenges. Instead, only certain topics are commonly addressed, while a number of issues such as anti-corruption measures or labour rights are underrepresented in the case study sample. Regarding the target regions of the best practice examples, the majority is reported on activities based in OECD countries and a small number of emerging markets such as South Africa, India or China, while neglecting other regions such as sub-Saharan Africa (excluding South Africa). From a European Union policy perspective, these results indicate that there is a role to play for the state in order to create a better fit between CSR agendas and the actual developmental needs in the South.