Long-term acclimation of leaf production, development, longevity and quality following 3 yr exposure to free-air CO2 enrichment during canopy closure in Populus

The effects of elevated CO2 on leaf development in three genotypes of Populus were investigated during canopy closure, following exposure to elevated CO2 over 3 yr using free-air enrichment.• Leaf quality was altered such that nitrogen concentration per unit d. wt (Nmass) declined on average by 22 and 13% for sun and shade leaves, respectively, in elevated CO2. There was little evidence that this was the result of ‘dilution’ following accumulation of nonstructural carbohydrates. Most likely, this was the result of increased leaf thickness. Specific leaf area declined in elevated CO2 on average by 29 and 5% for sun and shade leaves, respectively.• Autumnal senescence was delayed in elevated CO2 with a 10% increase in the number of days at which 50% leaf loss occurred in elevated as compared with ambient CO2.• These data suggest that changes in leaf quality may be predicted following long-term acclimation of fast-growing forest trees to elevated CO2, and that canopy longevity may increase, with important implications for forest productivity.

Human red blood cells at work: identification and visualization of erythrocytic eNOS activity in health and disease

A nitric oxide synthase (NOS)-like activity has been demonstrated in human red blood cells (RBCs), but doubts about its functional significance, isoform identity and disease relevance remain. Using flow cytometry in combination with the NO-imaging probe DAF-FM we find that all blood cells form NO intracellularly, with a rank order of monocytes > neutrophils > lymphocytes > RBCs > platelets. The observation of a NO-related fluorescence within RBCs was unexpected given the abundance of the NO-scavenger oxyhemoglobin. Constitutive normoxic NO formation was abolished by NOS inhibition and intracellular NO scavenging, confirmed by laser-scanning microscopy and unequivocally validated by detection of the DAF-FM reaction product with NO using HPLC and LC-MS/MS. Employing immunoprecipitation, ESI-MS/MS-based peptide sequencing and enzymatic assay we further demonstrate that human RBCs contain an endothelial NOS (eNOS) that converts L-3H-Arginine to L-3H-Citrulline in a Ca2+/Calmodulin-dependent fashion. Moreover, in patients with coronary artery disease, red cell eNOS expression and activity are both lower than in age-matched healthy individuals and correlate with the degree of endothelial dysfunction. Thus, human RBCs constitutively produce NO under normoxic conditions via an active eNOS isoform the activity of which is compromised in patients with coronary artery disease.

A multi-level analytical approach for detection and visualization of intracellular NO production and nitrosation events using diaminofluoresceins

Diaminofluoresceins are widely used probes for detection and intracellular localization of NO formation in cultured/isolated cells and intact tissues. The fluorinated derivative, 4-amino-5-methylamino-2′,7′-difluorofluorescein (DAF-FM), has gained increasing popularity in recent years due to its improved NO-sensitivity, pH-stability, and resistance to photo-bleaching compared to the first-generation compound, DAF-2. Detection of NO production by either reagent relies on conversion of the parent compound into a fluorescent triazole, DAF-FM-T and DAF-2-T, respectively. While this reaction is specific for NO and/or reactive nitrosating species, it is also affected by the presence of oxidants/antioxidants. Moreover, the reaction with other molecules can lead to the formation of fluorescent products other than the expected triazole. Thus additional controls and structural confirmation of the reaction products are essential. Using human red blood cells as an exemplary cellular system we here describe robust protocols for the analysis of intracellular DAF-FM-T formation using an array of fluorescence-based methods (laser-scanning fluorescence microscopy, flow cytometry and fluorimetry) and analytical separation techniques (reversed-phase HPLC and LC-MS/MS). When used in combination, these assays afford unequivocal identification of the fluorescent signal as being derived from NO and are applicable to most other cellular systems without or with only minor modifications.

Arctic ozone depletion in 2002–2003 measured by ASUR and comparison with POAM observations

We present ozone loss estimated from airborne measurements taken during January–February and March in the Arctic winter 2002/2003. The first half of the winter was characterized by unusually cold temperatures and the second half by a major stratospheric sudden warming around 15–18 January 2003. The potential vorticity maps show a vortex split in the lower stratosphere during the major warming (MW) in late January and during the minor warming in mid-February due to wave 1 amplification. However, the warming can be termed as a vortex displacement event as there was no vortex split during the MW period at 10 hPa. Very low temperatures, large areas of polar stratospheric clouds (PSCs), and high chlorine activation triggered significant ozone loss in the early winter, as the vortex moved to the midlatitude regions. The ozone depletion derived from the ASUR measurements sampled inside the vortex, in conjunction with the Mimosa-Chim model tracer, shows a maximum of 1.3 ± 0.2 ppmv at 450–500 K by late March. The partial column loss derived from the ASUR ozone profiles reaches up to 61 ± 4 DU in 400–550 K in the same period. The evolution of ozone and ozone loss assessed from the ASUR measurements is in very good agreement with POAM observations. The reduction in ozone estimated from the POAM measurements shows a similar maximum of 1.3 ± 0.2 ppmv at 400–500 K or 63 ± 4 DU in 400–550 K in late March. Our study reveals that the Arctic winter 2002/2003 was unique as it had three minor warmings and a MW, yet showed large loss in ozone. No such feature was observed in any other Arctic winter in the 1989–2010 period. In addition, an unusually large ozone loss in December, around 0.5 ± 0.2 ppmv at 450–500 K or 12 ± 1 DU in 400–550 K, was estimated for the first time in the Arctic. A careful and detailed diagnosis with all available published results for this winter exhibits an average ozone loss of 1.5 ± 0.3 ppmv at 450–500 K or 65 ± 5 DU in 400–550 K by the end of March, which exactly matches the ozone depletion derived from the ASUR, POAM and model data. The early ozone loss together with considerable loss afterwards put the warm Arctic winter 2002/2003 amongst the moderately cold winters in terms of the significance of the ozone loss.

Retrofit city futures: visions for urban sustainability

This report (which is part of the EPSRC Retrofit 2050 project) sets out three contrasting long term (2050) visions for retrofit city-regional futures, developed through an in-depth participatory backcasting and foresight process. These contextual scenarios are intended as a tool which can be adapted and used by a wide variety of stakeholders and organisations to stimulate discussion and inform future policy and long-term planning.

City futures: exploring urban retrofit and sustainable transitions

Cities are responsible for up to 70% of global carbon emissions and 75% of global energy consumption. By 2050 it is estimated that 70% of the world's population will live in cities. The critical challenge for contemporary urbanism, therefore, is to understand how to develop the knowledge, capacity and capability for public agencies, the private sector and multiple users in city-regions (i.e. the city and its wider hinterland) to re-engineer systemically their built environment and urban infrastructure in response to climate change and resource constraints. To inform transitions to urban sustainability, key stakeholders' perceptions were sought though a participatory backcasting and scenario foresight process in order to illuminate challenging but realistic socio-technical scenarios for the systemic retrofit of core UK city-regions. The challenge of conceptualizing complex urban transitions is explored across multiple socio-technical ‘regimes’ (housing, non-domestic buildings, urban infrastructure), scales (building, neighbourhood, city-region), and domains (energy, water, use of resources) within a participatory process. The development of three archetypal ‘guiding visions’ of retrofit city-regional futures developed through this process are discussed, along with the contribution that such foresight processes might play in ‘opening up’ the governance and strategic navigation of urban sustainability.

Patterning and detailed study of human hNT astrocytes on parylene-C/silicon dioxide substrates to the single cell level

It is estimated that the adult human brain contains 100 billion neurons with 5–10 times as many astrocytes. Although it has been generally considered that the astrocyte is a simple supportive cell to the neuron, recent research has revealed new functionality of the astrocyte in the form of information transfer to neurons of the brain. In our previous work we developed a protocol to pattern the hNT neuron (derived from the human teratocarcinoma cell line (hNT)) on parylene-C/SiO2 substrates. In this work, we report how we have managed to pattern hNT astrocytes, on parylene-C/SiO2 substrates to single cell resolution. This article disseminates the nanofabrication and cell culturing steps necessary for the patterning of such cells. In addition, it reports the necessary strip lengths and strip width dimensions of parylene-C that encourage high degrees of cellular coverage and single cell isolation for this cell type. The significance in patterning the hNT astrocyte on silicon chip is that it will help enable single cell and network studies into the undiscovered functionality of this interesting cell, thus, contributing to closer pathological studies of the human brain.

Low cost, patterning of human hNT brain cells on parylene-C with UV & IR laser machining

This paper describes the use of 800nm femtosecond infrared (IR) and 248nm nanosecond ultraviolet (UV) laser radiation in performing ablative micromachining of parylene-C on SiO2 substrates for the patterning of human hNT astrocytes. Results are presented that support the validity of using IR laser ablative micromachining for patterning human hNT astrocytes cells while UV laser radiation produces photo-oxidation of the parylene-C and destroys cell patterning. The findings demonstrate how IR laser ablative micromachining of parylene-C on SiO2 substrates can offer a low cost, accessible alternative for rapid prototyping, high yield cell patterning.

Urban retrofitting: identifying disruptive and sustaining technologies using performative and foresight techniques

Cities, which are now inhabited by a majority of the world's population, are not only an important source of global environmental and resource depletion problems, but can also act as important centres of technological innovation and social learning in the continuing quest for a low carbon future. Planning and managing large-scale transitions in cities to deal with these pressures require an understanding of urban retrofitting at city scale. In this context performative techniques (such as backcasting and roadmapping) can provide valuable tools for helping cities develop a strategic view of the future. However, it is also important to identify ‘disruptive’ and ‘sustaining’ technologies which may contribute to city-based sustainability transitions. This paper presents research findings from the EPSRC Retrofit 2050 project, and explores the relationship between technology roadmaps and transition theory literature, highlighting the research gaps at urban/city level. The paper develops a research methodology to describe the development of three guiding visions for city-regional retrofit futures, and identifies key sustaining and disruptive technologies at city scale within these visions using foresight (horizon scanning) techniques. The implications of the research for city-based transition studies and related methodologies are discussed.

Femininity, feminism and the terrorist

This chapter aims to discuss the relationship between femininity and representations of women involved in violence, focussing on visual representations. Miranda Alison has made the point that the repeated necessity to qualify the term 'combatant' with the descriptor 'female' draws attention to how women soldiers, female freedom fighters, female suicide bombers and female terrorists are exceptional figures. That the female combatant or the female terrorist is an aberration or a deviation from a masculine norm is undermined by the lengthy history of women as warriors, fighters, and terrorists. In that sense it is not so much that fighting women are rare but that there is amnesia within cultural memories concerning the woman fighter. However, in representations of conflict, the dominant image associated with femininity is passive; that is as the defenceless and the defended, or as the allegory of peace. Moreover, representations of men in wars as defeated or wounded means feminising such figures. Miriam Cooke, in her Women and the War Story, 1996, points out how a mythic war story provides men with political roles, in the politikon or public arena, whereas women are domesticated in the space of the oikon. In the mythic war story women may function as Mater Dolorosa, Patriotic Mother or Spartan Mother. It follows then that there are conditions in which it is permissible to represent women fighting on behalf of their children or in defence of the home, and in the absence of men. These images are also found in wider culture: Sarah Connor in Terminator or Ripley in Alien, for example. Images of the female terrorist raise new issues but I want to argue that it is also the case that discussing femininity and the terrorist must involve relating such imagery to representations of the female warrior over a longer timespan. Some questions have shifted since the late twentieth century. Dating from the early 1990s, most Western nations increasingly incorporated women into combat roles within their armed forces. This paper will aim to unpick some of the intricate connections between the increasing presence of women in the armed forces, what relationship this has to emancipation and the participation of women in violence classed as terrorist.