Translation of the flavivirus Kunjin NS3 gene in cis but not its RNA sequence or secondary structure is essential for efficient RNA packaging

Our previous studies using trans-complementation analysis of Kunjin virus (KUN) full-length cDNA clones harboring in-frame deletions in the NS3 gene demonstrated the inability of these defective complemented RNAs to be packaged into virus particles (W. J. Liu, P. L. Sedlak, N. Kondratieva, and A. A. Khromykh, J. Virol. 76:10766-10775). In this study we aimed to establish whether this requirement for NS3 in RNA packaging is determined by the secondary RNA structure of the NS3 gene or by the essential role of the translated NS3 gene product. Multiple silent mutations of three computer-predicted stable RNA structures in the NS3 coding region of KUN replicon RNA aimed at disrupting RNA secondary structure without affecting amino acid sequence did not affect RNA replication and packaging into virus-like particles in the packaging cell line, thus demonstrating that the predicted conserved RNA structures in the NS3 gene do not play a role in RNA replication and/or packaging. In contrast, double frameshift mutations in the NS3 coding region of full-length KUN RNA, producing scrambled NS3 protein but retaining secondary RNA structure, resulted in the loss of ability of these defective RNAs to be packaged into virus particles in complementation experiments in KUN replicon-expressing cells. Furthermore, the more robust complementation-packaging system based on established stable cell lines producing large amounts of complemented replicating NS3-deficient replicon RNAs and infection with KUN virus to provide structural proteins also failed to detect any secreted virus-like particles containing packaged NS3-deficient replicon RNAs. These results have now firmly established the requirement of KUN NS3 protein translated in cis for genome packaging into virus particles.

English collocation studies:the OSTI report

This is the first published edition of John Sinclair, Susan Jones and Robert Daley's research on collocation undertaken in 1970. The unpublished report was circulated amongst a small group of academics and was enormously influential, sparking a growth of interest in collocation amongst researchers in linguistics. Collocation was first viewed as important in computational linguistics in the work of Harold Palmer in Japan. Later M.A.K. Halliday and John Sinclair published on collocation in the 1960s. English Collocation Studies is a report on empirical research into collocation, devised by Halliday with Sinclair acting as the Principal Investigator and editor of the resultant OSTI report. The present edition contains an introduction by Professor Wolfgang Teubert based on his interview with John Sinclair. The introduction assesses the extent to which the findings of the original research have developed in the intervening years, and how some of the techniques mentioned in the report were implemented in the COBUILD project at Birmingham University in the 1980s.

Experimental and theoretical studies on a distributed temperature sensor based on Brillouin scattering

Results are reported from recent research on the use of the Brillouin gain/loss mechanism for distributed sensing. A theoretical model of the interaction of the pulsed and CW beams is described and compared with experiments. Results from a system with a 51 km sensing length are presented. We finally investigate issues related to the variation within the sensing fiber of the polarizations of the two beams.

Translation practices at the European Central Bank with reference to metaphors

The European Union institutions represent a complex setting and a specific case of institutional translation. The European Central Bank (ECB) is a particular context as the documents translated belong to the field of economics and, thus, contain many specialised terms and neologisms that pose challenges to translators. This study aims to investigate the translation practices at the ECB, and to analyse their effects on the translated texts. In order to illustrate the way texts are translated at the ECB, the thesis will focus on metaphorical expressions and the conceptual metaphors by which they are sanctioned. Metaphor is often associated with literature and less with specialised texts. However, according to Lakoff and Johnson’s (1980) conceptual metaphor theory, our conceptual system is fundamentally metaphorical in nature and metaphors are pervasive elements of thought and speech. The corpus compiled comprises economic documents translated at the ECB, mainly from English into Romanian. Using corpus analysis, the most salient metaphorical expressions were identified in the source and target texts and explained with reference to the main conceptual metaphors. Translation strategies are discussed on the basis of a comparison of the source and target texts. The text-based analysis is complemented by questionnaires distributed to translators, which give insights into the institution’s translation practices. As translation is an institutional process, translators have to follow certain guidelines and practices; these are discussed with reference to translators’ agency. A gap was identified in the field of institutional translation. The translation process in the EU institutions has been insufficiently explored, especially regarding the new languages of the European Union. By combining the analysis of the institutional practices, the texts produced in the institution and the translators’ work (by the questionnaires distributed to translators), this thesis intends to bring a contribution to institutional translation and metaphor translation, particularly regarding a new EU language, Romanian.

A comparative study of cationic liposome and niosome-based adjuvant systems for protein subunit vaccines:characterisation, environmental scanning electron microscopy and immunisation studies in mice

Vesicular adjuvant systems composing dimethyldioctadecylammonium (DDA) can promote both cell-mediated and humoral immune responses to the tuberculosis vaccine fusion protein in mice. However, these DDA preparations were found to be physically unstable, forming aggregates under ambient storage conditions. Therefore there is a need to improve the stability of such systems without undermining their potent adjuvanticity. To this end, the effect of incorporating non-ionic surfactants, such as 1-monopalmitoyl glycerol (MP), in addition to cholesterol (Chol) and trehalose 6,6′-dibehenate (TDB), on the stability and efficacy of these vaccine delivery systems was investigated. Differential scanning calorimetry revealed a reduction in the phase transition temperature (T c) of DDA-based vesicles by ∼12°C when MP and cholesterol (1:1 molar ratio) were incorporated into the DDA system. Transmission electron microscopy (TEM) revealed the addition of MP to DDA vesicles resulted in the formation of multi-lamellar vesicles. Environmental scanning electron microscopy (ESEM) of MP-Chol-DDA-TDB (16:16:4:0.5 μmol) indicated that incorporation of antigen led to increased stability of the vesicles, perhaps as a result of the antigen embedding within the vesicle bilayers. At 4°C DDA liposomes showed significant vesicle aggregation after 28 days, although addition of MP-Chol or TDB was shown to inhibit this instability. Alternatively, at 25°C only the MP-based systems retained their original size. The presence of MP within the vesicle formulation was also shown to promote a sustained release of antigen in-vitro. The adjuvant activity of various systems was tested in mice against three subunit antigens, including mycobacterial fusion protein Ag85b-ESAT-6, and two malarial antigens (Merozoite surface protein 1, MSP1, and the glutamate rich protein, GLURP). The MP- and DDA-based systems induced antibody responses at comparable levels whereas the DDA-based systems induced more powerful cell-mediated immune responses. © 2006 The Authors.

Saccharomyces Cerevisiae as a biotechnological tool for ageing research:studies on translation and metabolism

The yeast Saccharomyces cerevisiae is an important model organism for the study of cell biology. The similarity between yeast and human genes and the conservation of fundamental pathways means it can be used to investigate characteristics of healthy and diseased cells throughout the lifespan. Yeast is an equally important biotechnological tool that has long been the organism of choice for the production of alcoholic beverages, bread and a large variety of industrial products. For example, yeast is used to manufacture biofuels, lubricants, detergents, industrial enzymes, food additives and pharmaceuticals such as anti-parasitics, anti-cancer compounds, hormones (including insulin), vaccines and nutraceuticals. Its function as a cell factory is possible because of the speed with which it can be grown to high cell yields, the knowledge that it is generally recognized as safe (GRAS) and the ease with which metabolism and cellular pathways, such as translation can be manipulated. In this thesis, these two pathways are explored in the context of their biotechnological application to ageing research: (i) understanding translational processes during the high-yielding production of membrane protein drug targets and (ii) the manipulation of yeast metabolism to study the molecule, L-carnosine, which has been proposed to have anti-ageing properties. In the first of these themes, the yeast strains, spt3?, srb5?, gcn5? and yTHCBMS1, were examined since they have been previously demonstrated to dramatically increase the yields of a target membrane protein (the aquaporin, Fps1) compared to wild-type cells. The mechanisms underlying this discovery were therefore investigated. All high yielding strains were shown to have an altered translational state (mostly characterised by an initiation block) and constitutive phosphorylation of the translational initiation factor, eIF2a. The relevance of the initiation block was further supported by the finding that other strains, with known initiation blocks, are also high yielding for Fps1. A correlation in all strains between increased Fps1 yields and increased production of the transcriptional activator protein, Gcn4, suggested that yields are subject to translational control. Analysis of the 5´ untranslated region (UTR) of FPS1 revealed two upstream open reading frames (uORFs). Mutagenesis data suggest that high yielding strains may circumvent these control elements through either a leaky scanning or a re-initiation mechanism. In the second theme, the dipeptide L-carnosine (ß-alanyl-L-histidine) was investigated: it has previously been shown to inhibit the growth of cancer cells but delay senescence in cultured human fibroblasts and extend the lifespan of male fruit flies. To understand these apparently contradictory properties, the effects of L-carnosine on yeast were studied. S. cerevisiae can respire aerobically when grown on a non-fermentable carbon source as a substrate but has a respiro-fermentative metabolism when grown on a fermentable carbon source; these metabolisms mimic normal cell and cancerous cell metabolisms, respectively. When yeast were grown on fermentable carbon sources, in the presence of L-carnosine, a reduction in cell growth and viability was observed, which was not apparent for cells grown on a non-fermentable carbon source. The metabolism-dependent mechanism was confirmed in the respiratory yeast species Pichia pastoris. Further analysis of S. cerevisiae yeast strains with deletions in their nutrient-sensing pathway, which result in an increase in respiratory metabolism, confirmed the metabolism-dependent effects of L-carnosine.

Experimental and theoretical studies on a distributed temperature sensor based on Brillouin scattering

Results are reported from recent research on the use of the Brillouin gain/loss mechanism for distributed sensing. A theoretical model of the interaction of the pulsed and CW beams is described and compared with experiments. Results from a system with a 51 km sensing length are presented. We finally investigate issues related to the variation within the sensing fiber of the polarizations of the two beams.

Experimental and theoretical studies on a distributed temperature sensor based on Brillouin scattering

Results are reported from recent research on the use of the Brillouin gain/loss mechanism for distributed sensing. A theoretical model of the interaction of the pulsed and CW beams is described and compared with experiments. Results from a system with a 51 km sensing length are presented. We finally investigate issues related to the variation within the sensing fiber of the polarizations of the two beams.

The role of social agents in the translation into English of the novels of Naguib Mahfouz

This research investigates the field of translation in an Egyptain context around the work of the Egyptian writer and Nobel Laureate Naguib Mahfouz by adopting Pierre Bourdieu’s sociological framework. Bourdieu’s framework is used to examine the relationship between the field of cultural production and its social agents. The thesis includes investigation in two areas: first, the role of social agents in structuring and restructuring the field of translation, taking Mahfouz’s works as a case study; their role in the production and reception of translations and their practices in the field; and second, the way the field, with its political and socio-cultural factors, has influenced translators’ behaviour and structured their practices. In this research, it is argued that there are important social agents who have contributed significantly to the structure of the field and its boundaries. These are key social agents in the field namely; the main English language publisher in Egypt, the American University in Cairo Press (AUCP); the translators: Denys Johnson-Davies, Roger Allen and Trevor Le Gassick; and the author, Naguib Mahfouz. Their roles and contributions are examined and discussed through the lens of Bourdieu’s sociology. Particular focus is given to the author Mahfouz and his award of the Nobel Prize, and how this award has influenced the field of cultural production and its social agents. Also, it is argued that socio-cultural factors in the field, in the period between 1960s and 2000s, affected the translators’ practices in terms of modes of production of Mahfouz’s works. To investigate the influence of these factors on translators’ practices in the field, empirical examination is conducted, at the textual level, on a corpus of six translated novels written by the same author, Mahfouz. It is shown that the translators have an increased tendency, over time, towards applying a foreignising approach in their translations of culture-specific items. The translators’ behaviour, which is a result of their habitus, is correlated to political and socio-cultural factors in the field of translation. That is, based on interviews conducted with the translators, it has been found that there are particular factors influenced their translational habitus and, thus, their practices during the production process of the translations.

The translation of cell-based therapies:clinical landscape and manufacturing challenges

Cell-based therapies have the potential to make a large contribution toward currently unmet patient need and thus effective manufacture of these products is essential. Many challenges must be overcome before this can become a reality and a better definition of the manufacturing requirements for cell-based products must be obtained. The aim of this study is to inform industry and academia of current cell-based therapy clinical development and to identify gaps in their manufacturing requirements. A total of 1342 active cell-based therapy clinical trials have been identified and characterized based on cell type, target indication and trial phase. Multiple technologies have been assessed for the manufacture of these cell types in order to facilitate product translation and future process development.

The design of contact lens based ocular drug delivery systems for single-day use:part (I) structural factors, surrogate ophthalmic dyes and passive diffusion studies

The poor retention and efficacy of instilled drops as a means of delivering drugs to the ophthalmic environment is well-recognised. The potential value of contact lenses as a means of ophthalmic drug delivery, and consequent improvement of pre-corneal retention is one obvious route to the development of a more effective ocular delivery system. Furthermore, the increasing availability and clinical use of daily disposable contact lenses provides the platform for the development of viable single-day use drug delivery devices based on existing materials and lenses. In order to provide a basis for the effective design of such devices, a systematic understanding of the factors affecting the interaction of individual drugs with the lens matrix is required. Because a large number of potential structural variables are involved, it is necessary to achieve some rationalisation of the parameters and physicochemical properties (such as molecular weight, charge, partition coefficients) that influence drug interactions. Ophthalmic dyes and structurally related compounds based on the same core structure were used to investigate these various factors and the way in which they can be used in concert to design effective release systems for structurally different drugs. Initial studies of passive diffusional release form a necessary precursor to the investigation of the features of the ocular environment that over-ride this simple behaviour. Commercially available contact lenses of differing structural classifications were used to study factors affecting the uptake of the surrogate actives and their release under 'passive' conditions. The interaction between active and lens material shows considerable and complex structure dependence, which is not simply related to equilibrium water content. The structure of the polymer matrix itself was found to have the dominant controlling influence on active uptake; hydrophobic interaction with the ophthalmic dye playing a major role. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

Noun Sense Disambiguation using Co-Occurrence Relation in Machine Translation

Word Sense Disambiguation, the process of identifying the meaning of a word in a sentence when the word has multiple meanings, is a critical problem of machine translation. It is generally very difficult to select the correct meaning of a word in a sentence, especially when the syntactical difference between the source and target language is big, e.g., English-Korean machine translation. To achieve a high level of accuracy of noun sense selection in machine translation, we introduced a statistical method based on co-occurrence relation of words in sentences and applied it to the English-Korean machine translator RyongNamSan. ACM Computing Classification System (1998): I.2.7.