Language and the Production of Space

This paper is a critical investigation into the relationship between language and spatial production within the context of post war social housing in the UK. Adopting an obsolete council estate in north London as an exemplar of British social housing of this period, this paper provides a critical reflection on a collaborative arts project titled 'The Market Estate Project' produced in 2010, just prior to the estates demolition. Coordinated by artists collaborative Tall Tales, backed by the Southern Housing Association and financed by the British Arts Council, 'The Market Estate Project' called for site specific artworks that celebrated ‘a colorful and rich last memory of the estate’, with an emphasis on collaboration and participation with residents and the local community. Combining theoretical research and site specific intervention, this paper explores the notion of ‘effective historical analysis’, in which the singularities pertaining to the transformation of language of social housing reveals parallels between the forces and networks which govern its transformations. The imagery presented within the paper is the documentation of practice led research comprising of two large-scale site specific interventions. The first project titled ‘Embodied Text’ (2010) was installed in the corridor of the Market Estate and explores the relationship between language, the body and the city. The second project titled ‘Rise and Fall’ (2010), is an investigation into the relationship between social and architectural space and involves the reproduction of a council estate flat reconfigured within a white cube gallery. Through an investigation of the complex networks, multiple logics and rich contradictions inherent in the field of social housing, this research suggests that language produces space and conversely space produces language echoing Henri Lefebvre’s proposition ‘that space and the political organization of space, expresses social relationships but also react back upon them.

Development of a minigenome cassette for Lettuce necrotic yellows virus: A first step in rescuing a plant cytorhabdovirus

Rhabdoviruses are enveloped negative-sense RNA viruses that have numerous biotechnological applications. However, recovering plant rhabdoviruses from cDNA remains difficult due to technical difficulties such as the need for concurrent in planta expression of the viral genome together with the viral nucleoprotein (N), phosphoprotein (P) and RNA-dependent RNA polymerase (L) and viral genome instability in E. coli. Here, we developed a negative-sense minigenome cassette for Lettuce necrotic yellows virus (LNYV). We introduced introns into the unstable viral ORF and employed Agrobacterium tumefaciens to co-infiltrate Nicotiana with the genes for the N, P, and L proteins together with the minigenome cassette. The minigenome cassette included the Discosoma sp. red fluorescent protein gene (DsRed) cloned in the negative-sense between the viral trailer and leader sequences which were placed between hammerhead and hepatitis delta ribozymes. In planta DsRed expression was demonstrated by western blotting while the appropriate splicing of introduced introns was confirmed by sequencing of RT-PCR product

Antibody degradation in tobacco plants: a predominantly apoplastic process.

BACKGROUND: Interest in using plants for production of recombinant proteins such as monoclonal antibodies is growing, but proteolytic degradation, leading to a loss of functionality and complications in downstream purification, is still a serious problem. RESULTS: In this study, we investigated the dynamics of the assembly and breakdown of a human IgG(1)κ antibody expressed in plants. Initial studies in a human IgG transgenic plant line suggested that IgG fragments were present prior to extraction. Indeed, when the proteolytic activity of non-transgenic Nicotiana tabacum leaf extracts was tested against a human IgG1 substrate, little activity was detectable in extraction buffers with pH > 5. Significant degradation was only observed when the plant extract was buffered below pH 5, but this proteolysis could be abrogated by addition of protease inhibitors. Pulse-chase analysis of IgG MAb transgenic plants also demonstrated that IgG assembly intermediates are present intracellularly and are not secreted, and indicates that the majority of proteolytic degradation occurs following secretion into the apoplastic space. CONCLUSIONS: The results provide evidence that proteolytic fragments derived from antibodies of the IgG subtype expressed in tobacco plants do not accumulate within the cell, and are instead likely to occur in the apoplastic space. Furthermore, any proteolytic activity due to the release of proteases from subcellular compartments during tissue disruption and extraction is not a major consideration under most commonly used extraction conditions

Sustainable Tourism Development in Rote Ndao

Located in the eastern province of Nusa Tenggara Timur (West Timor), Rote Ndao is an island of 120,000 residents that can be accessed via ferry from the West Timorese capital of Kupang. Prior to East Timorese independence in 2002, Kupung saw direct international flights from Australia, since redirected through Bali and Jakarta. Rote Ndao is traditionally focused on subsistence farming and the harvesting of the Lontar Palm; however, recent years have seen an influx of foreign interest as domestic and international investors buy up large tracks of beach front property. White sand beaches and world class surfing, Rote is a quintessential island paradise and many are predicting that this remote part of eastern Indonesia will become the new Bali of the 21st century

Staff unavailability and safe staffing: Are headroom allowances 'realistic'?

Background “Hours per patient day” (HPPD) is an internationally recognised resourcing metric used to measure direct nursing care hours. However, hospitals often under-estimate indirect time (unavailability) and specify unrealistic targets for planned unavailability (“headroom”). Aims To investigate the disparities between planned unavailability (“headroom”) and actual staff unavailability. Methods Data were collected from the e-rostering systems of 87 NHS Trusts. This was compared with published data from 35 roster policies. Results Many hospitals use headroom as a Key Performance Indicator (KPI) and set targets for its components in their roster policies. This research highlights large variations in unavailability (15.8% to 33.6%) and lower variations in headroom (16-26%). Conclusions Hospitals operationalise headroom around an idealised ‘target’ value. This may be detrimental. Compelling a unit with unavailability between 28-30%, to adopt an institution-wide headroom of, say, 22% may, at best, increase spending on bank/agency staff, or, at worse, jeopardise patient safety

Dynamic Analysis of Vascular Morphogenesis Using Transgenic Quail Embryos

Background: One of the least understood and most central questions confronting biologists is how initially simple clusters or sheet-like cell collectives can assemble into highly complex three-dimensional functional tissues and organs. Due to the limits of oxygen diffusion, blood vessels are an essential and ubiquitous presence in all amniote tissues and organs. Vasculogenesis, the de novo self-assembly of endothelial cell (EC) precursors into endothelial tubes, is the first step in blood vessel formation [1]. Static imaging and in vitro models are wholly inadequate to capture many aspects of vascular pattern formation in vivo, because vasculogenesis involves dynamic changes of the endothelial cells and of the forming blood vessels, in an embryo that is changing size and shape. Methodology/Principal Findings: We have generated Tie1 transgenic quail lines Tg(tie1:H2B-eYFP) that express H2B-eYFP in all of their endothelial cells which permit investigations into early embryonic vascular morphogenesis with unprecedented clarity and insight. By combining the power of molecular genetics with the elegance of dynamic imaging, we follow the precise patterning of endothelial cells in space and time. We show that during vasculogenesis within the vascular plexus, ECs move independently to form the rudiments of blood vessels, all while collectively moving with gastrulating tissues that flow toward the embryo midline. The aortae are a composite of somatic derived ECs forming its dorsal regions and the splanchnic derived ECs forming its ventral region. The ECs in the dorsal regions of the forming aortae exhibit variable mediolateral motions as they move rostrally; those in more ventral regions show significant lateral-to-medial movement as they course rostrally. Conclusions/Significance: The present results offer a powerful approach to the major challenge of studying the relative role(s) of the mechanical, molecular, and cellular mechanisms of vascular development. In past studies, the advantages of the molecular genetic tools available in mouse were counterbalanced by the limited experimental accessibility needed for imaging and perturbation studies. Avian embryos provide the needed accessibility, but few genetic resources. The creation of transgenic quail with labeled endothelia builds upon the important roles that avian embryos have played in previous studies of vascular development

The Art of Data Science

To flourish in the new data-intensive environment of 21st century science, we need to evolve new skills. These can be expressed in terms of the systemized framework that formed the basis of mediaeval education - the trivium (logic, grammar, and rhetoric) and quadrivium (arithmetic, geometry, music, and astronomy). However, rather than focusing on number, data is the new keystone. We need to understand what rules it obeys, how it is symbolized and communicated and what its relationship to physical space and time is. In this paper, we will review this understanding in terms of the technologies and processes that it requires. We contend that, at least, an appreciation of all these aspects is crucial to enable us to extract scientific information and knowledge from the data sets which threaten to engulf and overwhelm us.Comment: 12 pages, invited talk at Astrostatistics and Data Mining in Large Astronomical Databases workshop, La Palma, Spain, 30 May - 3 June 2011, to appear in Springer Series on Astrostatistic

Multiple gene expression in plants using MIDAS-P, a versatile type II restriction-based modular expression vector.

MIDAS-P is a plant expression vector with blue/white screening for iterative cloning of multiple, tandemly-arranged transcription units (TUs). We have used the MIDAS-P system to investigate expression of up to five genes encoding three anti-HIV proteins and the reporter gene DsRed in Nicotiana benthamiana plants. The anti-HIV cocktail was made up of a broadly neutralizing monoclonal antibody (VRC01), a lectin (Griffithsin), and a single-chain camelid nanobody (J3-VHH). Constructs containing different combinations of 3, 4 or 5 TUs encoding different components of the anti-HIV cocktail were assembled. mRNA levels of the genes of interest decreased beyond two TUs. Co-expression of the RNA silencing suppressor P19 dramatically increased overall mRNA and protein expression levels of each component. The position of individual TUs in 3 TU constructs did not affect mRNA or protein expression levels. However, their expression dropped to non-detectable levels in constructs with 4 or more TUs each containing the same promoter and terminator elements, with the exception of DsRed at the first or last position in 5 TU constructs. This drop was alleviated by co-expression of P19. In short, the MIDAS-P system is suitable for the simultaneous expression of multiple proteins in one construct. This article is protected by copyright. All rights reserved

Expression of Protease-Activated Receptor 1 and 2 and Anti-Tubulogenic Activity of Protease-Activated Receptor 1 in Human Endothelial Colony-Forming Cells

Endothelial colony-forming cells (ECFCs) are obtained from the culture of human peripheral blood mononuclear cell (hPBMNC) fractions and are characterised by high proliferative and pro-vasculogenic potential, which makes them of great interest for cell therapy. Here, we describe the detection of protease-activated receptor (PAR) 1 and 2 amongst the surface proteins expressed in ECFCs. Both receptors are functionally coupled to extracellular signal-regulated kinase (ERK) 1 and 2, which become activated and phosphorylated in response to selective PAR1- or PAR2-activating peptides. Specific stimulation of PAR1, but not PAR2, significantly inhibits capillary-like tube formation by ECFCs in vitro, suggesting that tubulogenesis is negatively regulated by proteases able to stimulate PAR1 (e.g. thrombin). The activation of ERKs is not involved in the regulation of tubulogenesis in vitro, as suggested by use of the MEK inhibitor PD98059 and by the fact that PAR2 stimulation activates ERKs without affecting capillary tube formation. Both qPCR and immunoblotting showed a significant downregulation of vascular endothelial growth factor 2 (VEGFR2) in response to PAR1 stimulation. Moreover, the addition of VEGF (50–100 ng/ml) but not basic Fibroblast Growth Factor (FGF) (25–100 ng/ml) rescued tube formation by ECFCs treated with PAR1-activating peptide. Therefore, we propose that reduction of VEGF responsiveness resulting from down-regulation of VEGFR2 is underlying the anti-tubulogenic effect of PAR1 activation. Although the role of PAR2 remains elusive, this study sheds new light on the regulation of the vasculogenic activity of ECFCs and suggests a potential link between adult vasculogenesis and the coagulation cascade

X-Ray Spectroscopy of Stars

(abridged) Non-degenerate stars of essentially all spectral classes are soft X-ray sources. Low-mass stars on the cooler part of the main sequence and their pre-main sequence predecessors define the dominant stellar population in the galaxy by number. Their X-ray spectra are reminiscent, in the broadest sense, of X-ray spectra from the solar corona. X-ray emission from cool stars is indeed ascribed to magnetically trapped hot gas analogous to the solar coronal plasma. Coronal structure, its thermal stratification and geometric extent can be interpreted based on various spectral diagnostics. New features have been identified in pre-main sequence stars; some of these may be related to accretion shocks on the stellar surface, fluorescence on circumstellar disks due to X-ray irradiation, or shock heating in stellar outflows. Massive, hot stars clearly dominate the interaction with the galactic interstellar medium: they are the main sources of ionizing radiation, mechanical energy and chemical enrichment in galaxies. High-energy emission permits to probe some of the most important processes at work in these stars, and put constraints on their most peculiar feature: the stellar wind. Here, we review recent advances in our understanding of cool and hot stars through the study of X-ray spectra, in particular high-resolution spectra now available from XMM-Newton and Chandra. We address issues related to coronal structure, flares, the composition of coronal plasma, X-ray production in accretion streams and outflows, X-rays from single OB-type stars, massive binaries, magnetic hot objects and evolved WR stars.Comment: accepted for Astron. Astrophys. Rev., 98 journal pages, 30 figures (partly multiple); some corrections made after proof stag