SARS coronavirus nucleocapsid immunodominant T-cell epitope cluster is common to both exogenous recombinant and endogenous DNA-encoded immunogens

Correspondence between the T-cell epitope responses of vaccine immunogens and those of pathogen antigens is critical to vaccine efficacy. In the present study, we analyzed the spectrum of immune responses of mice to three different forms of the SARS coronavirus nucleocapsid (N): (1) exogenous recombinant protein (N-GST) with Freund's adjuvant; (2) DNA encoding unmodified N as an endogenous cytoplasmic protein (pN); and (3) DNA encoding N as a LAMP-I chimera targeted to the lysosomal MHC II compartment (p-LAMP-N). Lysosomal trafficking of the LAMP/N chimera in transfected cells was documented by both confocal and immunoelectron microscopy. The responses of the immunized mice differed markedly. The strongest T-cell IFN-gamma and CTL responses were to the LAMP-N chimera followed by the pN immunogen. In contrast, N-GST elicited strong T cell IL-4 but minimal IFN-gamma responses and a much greater antibody response. Despite these differences, however, the immunodominant T-cell ELISpot responses to each of the three immunogens were elicited by the same N peptides, with the greatest responses being generated by a cluster of five overlapping peptides, N76-114, each of which contained nonameric H2(d) binding domains with high binding scores for both class I and, except for N76-93, class II alleles. These results demonstrate that processing and presentation of N, whether exogenously or endogenously derived, resulted in common immunodominant epitopes, supporting the usefulness of modified antigen delivery and trafficking forms and, in particular, LAMP chimeras as vaccine candidates. Nevertheless, the profiles of T-cell responses were distinctly different. The pronounced Th-2 and humoral response to N protein plus adjuvant are in contrast to the balanced IFN-gamma and IL-4 responses and strong memory CTL responses to the LAMP-N chimera. (C) 2005 Elsevier Inc. All rights reserved.

Control of culture environment for improved polyethylenimine-mediated transient production of recombinant monoclonal antibodies by CHO cells

In this study we describe optimization of polyethylenimine (PEI)-mediated transient production of recombinant protein by CHO cells by facile manipulation of a chemically defined culture environment to limit accumulation of nonproductive cell biomass, increase the duration of recombinant protein production from transfected plasmid DNA, and increase cell-specific production. The optimal conditions for transient transfection of suspension-adapted CHO cells using branched, 25 kDa PEI as a gene delivery vehicle were experimentally determined by production of secreted alkaline phosphatase reporter in static cultures and recombinant IgG(4) monoclonal antibody (Mab) production in agitated shake flask cultures to be a DNA concentration of 1.25 mu g 10(6) cells(-1) mL(-1) at a PEI nitrogen: DNA phosphate ratio of 20:1. These conditions represented the optimal compromise between PEI cytotoxicity and product yield with most efficient recombinant DNA utilization. Separately, both addition of recombinant insulin-like growth factor (LR3-IGF) and a reduction in culture temperature to 32 degrees C were found to increase product titer 2- and 3-fold, respectively. However, mild hypothermia and LR3-IGF acted synergistically to increase product titer 11-fold. Although increased product titer in the presence of LR3-IGF alone was solely a consequence of increased culture duration, a reduction in culture temperature post-transfection increased both the integral of viable cell concentration (IVC) and cell-specific Mab production rate. For cultures maintained at 32 degrees C in the presence of LR3-IGF, IVC and qMab were increased 4- and 2.5-fold, respectively. To further increase product yield from transfected DNA, the duration of transgene expression in cell populations maintained at 32 C in the presence of LR3-IGF was doubled by periodic resuspension of transfected cells in fresh media, leading to a 3-fold increase in accumulated Mab titer from similar to 13 to similar to 39 mg L-1. Under these conditions, Mab glycosylation at Asn297 remained essentially constant and similar to that of the same Mab produced by stably transfected GS-CHO cells. From these data we suggest that the efficiency of transient production processes (protein output per rDNA input) can be significantly improved using a combination of mild hypothermia and growth factor(s) to yield an extended activated hypothermic synthesis.

Response and function of cutaneous mucosal and serum antibodies in barramundi (Lates calcarifer) acclimated in seawater and freshwater

Mucosal and serum antibody responses were studied in sibling barramundi (Lates calcarifer) acclimated in either seawater or freshwater following vaccination by intraperitoneal injection or direct immersion in an inactivated Streptococcus iniae vaccine. As expected, route of vaccination had a marked effect on immune response, with direct immersion resulting in low serum antibody levels against S. iniae by ELISA detected 21 days post vaccination at 26 degrees C, whilst a significant response was detected in mucus. A strong specific antibody response was detected in both mucus and serum 21 days following intraperitoneal injection. Fish acclimated in seawater prior to vaccination showed a markedly higher specific mucosal antibody response than sibling fish acclimated in freshwater, regardless of the route of vaccination, whilst the serum antibody response was not affected by salinity. Both mucosal and serum antibodies from fish in seawater and freshwater were capable of binding antigen at salinities similar to full strength seawater in a modified ELISA assay. These results indicate that this euryhaline fish species is riot only able to mount significant specific antibody response in cutaneous mucus, but that these antibodies will function in the marine environment. (c) 2005 Elsevier Ltd. All rights reserved.

Equation of state of a superfluid Fermi gas in the BCS-BEC crossover

We present a theory for a superfluid Fermi gas near the BCS-BEC crossover, including pairing fluctuation contributions to the free energy similar to that considered by Nozieres and Schmitt-Rink for the normal phase. In the strong coupling limit, our theory is able to recover the Bogoliubov theory of a weakly interacting Bose gas with a molecular scattering length very close to the known exact result. We compare our results with recent Quantum Monte Carlo simulations both for the ground state and at finite temperature. Excellent agreement is found for all interaction strengths where simulation results are available.

Assembly of Human Papillomavirus Type-16 Virus-Like Particles: Multifactorial Study of Assembly and Competing Aggregation

Pentameric capsomeres of human papillomavirus capsid protein L1 expressed in Escherichia coli self-assemble into virus-like particles (VLPs) in vitro. A multifactorial experimental design was used to explore a wide range of solution conditions to optimize the assembly process. The degree of assembly was measured using an enzyme-linked immunosorbent assay, and a high-throughput turbidity assay was developed to monitor competing aggregation. The presence of zinc ions in the assembly buffer greatly increased the incidence of aggregation and had to be excluded from the experiment for meaningful analysis. Assembly of VLPs was optimal at a pH of about 6.5, calcium and sodium ions had no measurable effect, and dithiothreitol and glutathione inhibited assembly. Tryptophan fluorescence spectroscopy demonstrated that an increase in urea concentration reduced the rate of VLP formation but had no effect on the final concentration of assembled VLPs. This study demonstrates the use of the hanging-drop vapor-diffusion crystallization method to screen for conditions that promote aggregation and the use of tryptophan fluorescence spectroscopy for real-time monitoring of the assembly process.

Partitioning of deformation within a subduction channel during exhumation of high-pressure rocks: a case study from the Western Alps

The metamorphic belt of the Western Alps was subjected to widespread extensional tectonism at the end of the Eocene (ca. 45-35 Ma). Extension was accommodated by hinterland-directed movements along gently inclined extensional shear zones, which facilitated rapid exhumation of high-pressure and ultra-high-pressure rocks. This deformation resulted in a normal metamorphic sequence. Extension in the inner parts of the Western Alps was coeval with shortening at the front of the belt (foreland-directed thrusts), which took place during decompression, and emplaced higher grade metamorphic units over lower grade metamorphic rocks, thus forming an inverse metamorphic sequence. Two mechanisms for this extensional episode are discussed: (1) collapse of an overthickened lithosphere, and (2) internal readjustments within the orogenic wedge due to subduction channel dynamics. We favour the latter mechanism because it can account for the development of the observed inverse and normal metamorphic sequences along foreland-directed thrusts and hinterland-directed detachments, respectively. This hypothesis is supported by published structural, metamorphic and geochronological data from four geological transects through the Western Alps. This study also emphasizes the importance of post-shearing deformation (e.g. horizontal buckling versus vertical flattening), which can modify the distribution of hinterland- and foreland-directed shear zones in orogenic belts. (c) 2006 Elsevier Ltd. All rights reserved.

The other on display: translation in the Ethnographic Museum

The ethnographic museum in the West has a long and troubling history. The display of 'exotic peoples' in travelling exhibitions began as early as the sixteenth century, but it was the mid and late nineteenth century that saw the great expansion of museums as sites to show artefacts collected - under anything but reputable circumstances - from what were considered the 'primitive', 'natural', or 'tribal' peoples of the world. Today the ethnographic museum is still a feature of large European cities, though faced with newly formulated dilemmas in the postcolonial world. For how can the material culture of a non-western people be collected and displayed in the West without its makers being translated into wordless and powerless objects of visual consumption? In national museums the processes of choosing, contextualizing and commentating exhibits help form national identity; in the ethnographic museum, similarly, they shape perceptions of the apparently distant Other. Like written ethnography, the museum is a 'translation of culture', with many of the associated problems traced by Talal Asad (1986). Like the written form, it has to represent the dialogic realities of cultural encounters in a fixed and intelligible form, to propose categories that define and order the material it has gathered. As the public face of academic ethnography, the museum interprets other cultures for the benefit of the general reader, and in that task museum practice, like all ethnography, operates within very specific historical and political parameters. How are museums in western Europe responding to the issues raised by critical ethnographers like James Clifford (1988), with their focus on the politics of representation? Is globalisation increasing the degree of accountability imposed on the ethnographic museum, or merely reinforcing older patterns? What opportunities and problems are raised by the use of more words - more 'translation' in the narrower sense - in ethnographic museums, and how do museums gain from introducing a reflexive and contextualizing concept of "thick translation" (Appiah 1993) into their work of interpretation?

Representing others:translation, ethnography and museum

Cultural anthropology has always been dependent on translation as a textual practice, and it has often used 'translation' as a metaphor to describe ethnography's processes of interpretation and cross-cultural comparison. Questions of intelligibility and representation are central to both translation studies and ethnographic writing - as are the dilemmas of cultural distance or proximity, exoticism or appropriation. Similarly, recent work in museum studies discusses problems of representation that are raised by ethnographic museums as multimedia 'translations'. However, as yet there has been remarkably little interdisciplinary exchange: neither has translation studies kept up with the sophistication of anthropology's investigations of meaning, representation and 'culture' itself, nor have anthropology and museum studies often looked to translation studies for analyses of language difference or concrete methods of tracing translation practices.

Parallel texts in translation

Creating an appropriate translation often means adapting the target text (TT) to the text-typological conventions of the target culture. Such knowledge can be gained by a comparative analysis of parallel texts, i.e. L2 and L1 texts of equal informativity which have been produced in similar communicative situations. Some problems related to (cross-cultural) text-typological conventions and the role of parallel texts for describing translation strategies are described, as well as implications for teaching translation. The discussion is supported with examples of parallel texts that are representative of various genres, such as instruction manuals, international treaties and tourist brochures.

Third Ways and new centres - ideological unity or difference?

This chapter illustrates extratextual and intratextual aspects of ideology as related to translation with a case study, a policy document by Tony Blair and Gerhard Schröder, jointly published in English and German in June 1999. Textual features of the two language versions are compared and linked to the social contexts. Concepts and methods of critical discourse analysis and of descriptive and functionalist approaches to translation are applied for this purpose. In particular, reactions to the German text in Germany are explained with reference to the socio-political and ideological conditions of the text production, which was a case of parallel text production combined with translation. It is illustrated that decisions at the linguistic micro-level have had effects for a political party, reflected for example in the German Social Democratic Party debating its identity due to the textual treatment of ideological keywords. The subtle differences revealed in a comparative analysis of the two texts indicate the text producers' awareness of ideological phenomena in the respective cultures. Both texts thus serve as windows onto ideologies and political power relations in the contemporary world.

Representational modes of thinking employed by children aged thirteen and fourteen, and their relationship to performance in standardised tests of ability, measures of creativity and personality

This thesis proposes that despite many experimental studies of thinking, and the development of models of thinking, such as Bruner's (1966) enactive, iconic and symbolic developmental modes, the imagery and inner verbal strategies used by children need further investigation to establish a coherent, theoretical basis from which to create experimental curricula for direct improvement of those strategies. Five hundred and twenty-three first, second and third year comprehensive school children were tested on 'recall' imagery, using a modified Betts Imagery Test; and a test of dual-coding processes (Paivio, 1971, p.179), by the P/W Visual/Verbal Questionnaire, measuring 'applied imagery' and inner verbalising. Three lines of investigation were pursued: 1. An investigation a. of hypothetical representational strategy differences between boys and girls; and b. the extent to which strategies change with increasing age. 2. The second and third year children's use of representational processes, were taken separately and compared with performance measures of perception, field independence, creativity, self-sufficiency and self-concept. 3. The second and third year children were categorised into four dual-coding strategy groups: a. High Visual/High Verbal b. Low Visual/High Verbal c. High Visual/Low Verbal d. Low Visual/Low Verbal These groups were compared on the same performance measures. The main result indicates that: 1. A hierarchy of dual-coding strategy use can be identified that is significantly related (.01, Binomial Test) to success or failure in the performance measures: the High Visual/High Verbal group registering the highest scores, the Low Visual/High Verbal and High Visual/Low Verbal groups registering intermediate scores, and the Low Visual/Low Verbal group registering the lowest scores on the performance measures. Subsidiary results indicate that: 2. Boys' use of visual strategies declines, and of verbal strategies increases, with age; girls' recall imagery strategy increases with age. Educational implications from the main result are discussed, the establishment of experimental curricula proposed, and further research suggested.

Towards understanding promiscuity in multidrug efflux pumps

Drug export from cells is a major factor in the acquisition of cellular resistance to antimicrobial and cancer chemotherapy, and poses a significant threat to future clinical management of disease. Many of the proteins that catalyse drug efflux do so with remarkably low substrate specificity, a phenomenon known as multidrug transport. For these reasons we need a greater understanding of drug recognition and transport in multidrug pumps to inform research that attempts to circumvent their action. Structural and computational studies have been heralded as being great strides towards a full elucidation of multidrug recognition and transport. In this review we summarise these advances and ask how close we are to a molecular understanding of this remarkable phenomenon. © 2013 Elsevier Ltd.

Introduction : special issues on key debates in advertising material

Research on advertising has been conducted mainly from within disciplines such as marketing and reception studies. Within translation studies, research has focused more on comparative analysis of language pairs for intersystemic differences, and more recently on intercultural differences in product appeal and cultural values. Advertising as text type was most comprehensively used and developed within Western cultures, then 'exported' to other cultures. Initially the assumption was that the message could be translated and the product would sell on the basis of Western value-based market research, although this is now clearly shown not to be the case, as contributors to this volume all demonstrate through empirical studies. The introduction to this volume offers a comprehensive survey of work done in the field, both within and outside translation studies, outlines the use of semiotics as a suitable framework for research, and summarizes the key issues relating to the global vs. local debate.