Functional Analysis of Subunit e of the F\u3csub\u3e1\u3c/sub\u3eF\u3csub\u3eo\u3c/sub\u3e-ATP Synthase of the Yeast \u3cem\u3eSaccharomyces cerevisiae\u3c/em\u3e: Importance of the N-Terminal Membrane Anchor Region

Mitochondrial F1Fo-ATP synthase complexes do not exist as physically independent entities but rather form dimeric and possibly oligomeric complexes in the inner mitochondrial membrane. Stable dimerization of two F1Fo-monomeric complexes involves the physical association of two membrane-embedded Fo-sectors. Previously, formation of the ATP synthase dimeric-oligomeric network was demonstrated to play a critical role in modulating the morphology of the mitochondrial inner membrane. In Saccharomyces cerevisiae, subunit e (Su e) of the Fo-sector plays a central role in supporting ATP synthase dimerization. The Su e protein is anchored to the inner membrane via a hydrophobic region located at its N-terminal end. The hydrophilic C-terminal region of Su e resides in the intermembrane space and contains a conserved coiled-coil motif. In the present study, we focused on characterizing the importance of these regions for the function of Su e. We created a number of C-terminal-truncated derivatives of the Su e protein and expressed them in the Su e null yeast mutant. Mitochondria were isolated from the resulting transformant strains, and a number of functions of Su e were analyzed. Our results indicate that the N-terminal hydrophobic region plays important roles in the Su e-dependent processes of mitochondrial DNA maintenance, modulation of mitochondrial morphology, and stabilization of the dimer-specific Fo subunits, subunits g and k. Furthermore, we show that the C-terminal coiled-coil region of Su e functions to stabilize the dimeric form of detergent-solubilized ATP synthase complexes. Finally, we propose a model to explain how Su e supports the assembly of the ATP synthase dimers-oligomers in the mitochondrial membrane

Plenty More Fish in the Sea? A working paper on the legal issues related to fishing beyond maximum sustainable yield: A UK case study

Maximum Sustainable Yield (MSY) is about to become a centre stage issue. The EU now has a commitment that fishing will progressively be managed at levels that correspond to MSY, and setting of quotas will respect scientific advice. Where possible, this is to be attained by 2015, and all fish stocks will be managed at MSY by 2020 at the latest. What the term ‘maximum sustainable yield’ means will dominate the next seven years of fisheries discussions. MSY has two different interpretations. Scientists use the term to mean the ability of a fish stock to support a sustainable fishing industry. Lawyers use it when assessing the obligations of a country in respect of its sovereign fishing rights. This report focuses on the legal interpretation of MSY. The report’s findings are particularly important as the EU has consistently over-allocated quota and ignored scientific advice, with the result that 75% of EU fish stocks are overfished. The starting point of a thorough investigation of MSY is the nature of fishing rights themselves. Fisheries were traditionally open access and seen as being inexhaustible. The origins of this approach date at least as far back as the era of the Roman Empire, but even then interpretation by the Roman courts created some perverse outcomes. The principle of open access became established for the English fishery in the 12th century. Before then, it is likely that all viable tidal fisheries were granted by the Crown into private hands following the Norman conquest. Indeed, some of these private tidal fisheries still persist today. In the seventeenth century, the mare liberum (freedom of the seas) debate confirmed open access on the high seas, but the position for territorial waters (within the 3 mile limit) was more complex. Under the common law, fishers operated under the public right to fish which permitted open access and has come to be interpreted as being based on the principle of inexhaustibility of the stock. Where this myth of inexhaustibility has come from is difficult to tell. It is often attributed to the Victorian scientist Thomas Huxley because of his pronouncement on the fecundity of some fish species, but Huxley himself highlights fisheries which were vulnerable to over-fishing and recognises that, on the high seas, no regulatory mechanisms were available, so this aspect of his findings was theoretical. Moreover, the negative impacts of some fisheries were known to the legislators at least since the Middle Ages and it is tempting to posit that public authorities, in whatever form they manifested themselves, have always been aware that some fishing methods can cause significant harm. As it stands, the basic principle the courts still adhere to is that the right to fish is untrammelled, but whether this principle would withstand direct judicial scrutiny is questionable. The United Nations Convention on the Laws of the Sea (UNCLOS) changed everything. Coastal states now enjoy sovereign rights over a far larger area of sea than they did previously. UK territorial waters now extend to 12 nautical miles and its exclusive economic zone (EEZ) extends up to 200 nautical miles. UNCLOS recognises sovereign rights for fisheries but also imposes duties on coastal states to restore stocks to levels which can produce MSY within their EEZs. Sovereign fishing rights are not untrammelled; they are qualified by a duty to restore stocks and to preserve the marine environment. Similar obligations are contained in the Convention on Biodiversity (with a target for 2020) and by virtue of the World Summit for Sustainable Development (with a target of 2015 for depleted stocks). Increasingly, international obligations of this nature are being enforced through the domestic courts. Many countries (including the UK itself for its Crown Dependencies and overseas territories) vest their fishing rights in an identifiable legal entity which then not just regulates the fishery, but also acts as an owner in the way it disposes of the fishery to commercial operators. Unfortunately, for the waters immediately adjacent to the UK, it is not clear in what Crown entity the UK’s fishing rights actually vest. The UK fishery is clearly some form of public asset and it is likely that there is a Crown trust in existence. The terms of this trust would place similar duties to maintain and restore fish stocks as those contained within UNCLOS. Despite a first instance decision to the contrary, it is possible that the terms of that trust are enforceable by third parties. There is a well-established principle in English law of nemo dat quod non habet (you cannot dispose of something you don’t own). The fact that the UK’s sovereign rights are qualified to MSY mean that the UK cannot grant to its fishers (or fishers of other Member States operating in UK waters) untrammelled rights which it does not possess itself. The same principle applies to the EU Common Fisheries Policy; the UK cannot confer on the EU greater fishing rights than those acknowledged by UNCLOS. The scientific origins of the term MSY were in the work of Beverton and Holt and related to a theoretical maximum, where a capture fishery will sustain itself. The legal meaning of the term gives discretion to fisheries managers to establish a management system which uses the best available science to determine what actions needs to be taken. The legal interpretation of MSY is therefore much broader than the scientific one and has the following features: • it is constantly evolving relating to the best available science of the time; • it defines the limits of a coastal state’s sovereign rights; • it may relate to the setting of fishing quota (where calculations of scientific MSY are particularly predominant), but many obligations for the attainment of MSY may also be satisfied by the imposition of technical or area-based management techniques such as marine protected areas; • it provides the framework in which fisheries should be managed; and • it can be used to show where stocks or areas of sea are not being properly managed, and thus direct where management resources should be targeted. The introduction of the ecosystem approach to fisheries management means that legal MSY can now take into consideration a much wider range of services provided by the fishery in the calculation and management of that fishery. UNCLOS itself permits taking into consideration relevant environmental and economic factors, including the economic needs of coastal fishing communities, but the ecosystem approach takes that a stage further by assessing all the services provided by the fishery. This shifts the focus away from simply treating the fishery as a commercial resource, to ensure that management benefits a much broader constituency. In 2008, the environmental non-governmental organisation (NGO), WWF tried to take the European Commission to the European Court of Justice for failing to allocate cod quota according to scientific advice. However, WWF was unable to access the European court system because the ECJ refused access to NGOs; ECJ rules at the time did not grant an NGO the right to be heard in court. Those rules are under scrutiny once again because they seem incompatible with the Aarhus Convention, so this position may change shortly. There are also other legal forums for testing the point. The UK courts do not have the same access restrictions as the ECJ, so a case launched in the UK courts concerning UK fishing rights or the voting patterns of the UK Minister in the EU could come to trial. The former President of the International Tribunal for the Laws of the Sea (ITLOS), Rüdiger Wolfrum, recommended ITLOS as having suitable jurisdiction to ensure compliance with MSY requirements The legal regime has dramatically moved away from the impossibility of mare liberum and open access fisheries to the possibility of effective regulation. Clearer sovereignty over the sea and better technology informing enforceable law means the legal context of fisheries have changed fundamentally for the better. There is an understanding that fisheries are held on trust for future generations. Fishing to MSY and trusteeship amount to the same thing; the legal interpretation of coastal states’ rights and responsibilities under MSY and the obligations of a Crown trust are the same. The ecosystem approach provides the matrix to understand who the beneficiaries of that trust are and where management effort needs to be directed. All these pieces are now in place and the hard law is there at the edges to ensure that reform remains on track. In short there are two key points to be drawn from this report: • It is illegal for coastal states to permit fishing beyond MSY; and • It is highly likely that this law is enforceable through the court system.Waterloo Foundatio

Macrophages Are Required for Dendritic Cell Uptake of Respiratory Syncytial Virus from an Infected Epithelium

We have previously shown that the respiratory syncytial virus [RSV] can productively infect monocyte derived dendritic cells [MoDC] and remain dormant within the same cells for prolonged periods. It is therefore possible that infected dendritic cells act as a reservoir within the airways of individuals between annual epidemics. In the present study we explored the possibility that sub-epithelial DCs can be infected with RSV from differentiated bronchial epithelium and that in turn RSV from DCs can infect the epithelium. A dual co-culture model was established in which a differentiated primary airway epithelium on an Air Liquid Interface (ALI) was cultured on a transwell insert and MoDCs were subsequently added to the basolateral membrane of the insert. Further experiments were undertaken using a triple co-culture model in which in which macrophages were added to the apical surface of the differentiated epithelium. A modified RSV [rr-RSV] expressing a red fluorescent protein marker of replication was used to infect either the MoDCs or the differentiated epithelium and infection of the reciprocal cell type was assessed using confocal microscopy. Our data shows that primary epithelium became infected when rr-RSV infected MoDCs were introduced onto the basal surface of the transwell insert. MoDCs located beneath the epithelium did not become infected with virus from infected epithelial cells in the dual co-culture model. However when macrophages were present on the apical surface of the primary epithelium infection of the basal MoDCs occurred. Our data suggests that RSV infected dendritic cells readily transmit infection to epithelial cells even when they are located beneath the basal layer. However macrophages appear to be necessary for the transmission of infection from epithelial cells to basal dendritic cells

Gut microbiota-derived propionate reduces cancer cell proliferation in the liver

Peer reviewedPublisher PD

Why don't they accept Non-Invasive Ventilation? : Insight into the interpersonal perspectives of patients with MND.

"Objectives. Although non-invasive ventilation (NIV) can benefit survival and quality of life, it is rejected by a substantial proportion of people with motor neurone disease (MND). The aim of this study was to understand why some MND patients decline or withdraw from NIV. Method. Nine patients withMND(male = 7, mean age = 67 years) participated in this study. These patients, from a cohort of 35 patients who were offered NIV treatment to support respiratory muscle weakness, did not participate in NIV treatment when it was clinically appropriate. Semi-structured interviews and interpretative phenomenological analysis (IPA) were employed to explore these patient’s experience of MND and their thoughts and understanding of NIV treatment. Results. Using IPA, four themes were identified: preservation of the self, negative perceptions of NIV, negative experience with health care services, and not needing NIV. Further analysis identified the fundamental issue to be the maintenance of perceived self, which was interpreted to consist of the sense of autonomy, dignity, and quality of life. Conclusions. The findings indicate psychological reasons for disengagement with NIV. The threat to the self, the sense of loss of control, and negative views of NIV resulting from anxiety were more important to these patients than prolonging life in its current form. These findings suggest the importance of understanding the psychological dimension involved in decision-making regarding uptake of NIV and a need for sensitive holistic evaluation if NIV is declined.

Additive sustainability footprint: Rationale and pilot evaluation of a tool for assessing the sustainable use of PVC additives

© 2019 The Authors. Journal of Vinyl and Additive Technology published by Wiley Periodicals, Inc. on behalf of Society of Plastics Engineers. PVC compounds contain additives necessary for processing and stability, and to modify the plastic's properties. The Europe-wide VinylPlus® voluntary commitment includes a challenge to make progress toward sustainable use of additives. Additive Sustainability Footprint (ASF) was developed to assess sustainable use of additives across the whole societal life cycles of finished PVC articles, taking a risk-based approach rather than simplistic hazard assessment. ASF addresses impacts across six life cycle assessment (LCA) stages established by ISO Standard 14040, using the four System Conditions (sustainability principles) developed by The Natural Step (TNS) covering social as well as environmental factors. For each LCA stage/System Condition combination, seven generically similar questions cover negative impacts (many covered by existing tools and regulations) but also the additive's positive contributions to the sustainability of finished articles. Positive contributions include ethical sourcing, longevity of service life, low maintenance inputs, and recyclability. Answers to questions determine a score, which can be combined across the life cycle and with other additives. Testing on a generic EU PVC window profile supported ASF development and demonstrated applicability and potential benefits including use for sensitivity analysis of alternative additives from different geopolitical regions or from recycled as opposed to virgin sources. J. VINYL ADDIT. TECHNOL., 2019. © 2019 The Authors. Journal of Vinyl and Additive Technology published by Wiley Periodicals, Inc. on behalf of Society of Plastics Engineers

Potential of a cyclone prototype spacer to improve in vitro dry powder delivery

Copyright The Author(s) 2013. This article is published with open access at Springerlink.com. This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are creditedPurpose: Low inspiratory force in patients with lung disease is associated with poor deagglomeration and high throat deposition when using dry powder inhalers (DPIs). The potential of two reverse flow cyclone prototypes as spacers for commercial carrierbased DPIs was investigated. Methods: Cyclohaler®, Accuhaler® and Easyhaler® were tested with and without the spacers between 30-60 Lmin-1. Deposition of particles in the next generation impactor and within the devices was determined by high performance liquid chromatography. Results: Reduced induction port deposition of the emitted particles from the cyclones was observed due to the high retention of the drug within the spacers (e.g. salbutamol sulphate (SS): 67.89 ± 6.51 % at 30 Lmin-1 in Cheng 1). Fine particle fractions of aerosol as emitted from the cyclones were substantially higher than the DPIs alone. Moreover, the aerodynamic diameters of particles emitted from the cyclones were halved compared to the DPIs alone (e.g. SS from the Cyclohaler® at 4 kPa: 1.08 ± 0.05 μm vs. 3.00 ± 0.12 μm, with and without Cheng 2, respectively) and unaltered with increased flow rates. Conclusion: This work has shown the potential of employing a cyclone spacer for commercial carrier-based DPIs to improve inhaled drug delivery.Peer reviewe

Work Package Report 6: Shared, plural and cultural values of ecosystems – Summary

Key findings: Finding 1: Shared values resulting from deliberative, group-based valuation are different from individual values. Case study evidence suggests that they are more informed, considered, confident and reflective of participants’ deeper-held, transcendental values. Deliberated, group-based monetary values may be a better reflection of real welfare impacts than non-deliberated individual values, if derived through a carefully designed and managed process. Although more research is needed to expand the currently small evidence base on deliberative monetary methods, group deliberation has the potential to significantly enhance elicitation of values. Finding 2: The ethical, moral and justice dimensions of many environmental issues necessitate approaches that allow for the elicitation of shared and plural values. Key ethical concerns include: 1) providing a space and opportunity for people to identify values that they may find difficult to articulate (e.g. spiritual, identity); 2) recognising that some values cannot be traded without discussion and negotiation (e.g. the legal or felt rights of local people, intrinsic values of other species); and 3) understanding that it is often difficult to isolate valuation from decision-making processes because people feel there are strong ethical or moral issues at stake that need to be debated (e.g. the justice of the process, fairness in the distribution of benefits or disbenefits, responsibility, and issues of sustainability and future generations). Finding 3: Catalyst and/or conflict points can play a key role in the emergence and articulation of values at a societal or community level that have not previously been outwardly or explicitly articulated. Catalyst and conflict points can be symbolic and are often linked to wider contested issues and meanings about who is involved in decision-making, whose voice counts and who receives the benefits or disbenefits of environmental change. These catalyst points can potentially be connected to feelings of powerlessness that give rise to concern and protest. By recognising transcendental societal and communal values (the deeper-held and overarching values held by society and communities), it becomes possible to make these values explicit and incorporate them in decision-making to better anticipate and manage conflicts. Finding 4: There is a diversity of ways in which shared, plural, cultural and social values are used, but they are rarely conceptualised. The UK NEAFO provides a clear theoretical framework that distinguishes and categorises different dimensions and types of shared values. The proposed range of value types was both identifiable and distinguishable within case study results. This suggests that the framework provides a useful basis for operationalizing shared values for decision-making. Finding 5: Shared and social values in the sense of value to society is conceptualised very differently by conventional economics and other disciplines. Neoclassical economists have generally undertaken valuation by equating social value with the aggregate of individual values. They consider values as fundamentally commensurable. In contrast, literature from other disciplines consistently considers values as plural, not just in the sense that multiple things have value, but also that there are multiple dimensions to value that cannot necessarily be captured in a single metric. Within mainstream economics, the difficulties associated with commensurability and aggregating values have long been recognised, but have also been neglected. An interesting area for future debate between economic and non-economic views on values may be the normative nature of value-aggregation. Finding 6: A mixed method approach is required to elicit the multiple dimensions of shared values and to translate deeper-held, transcendental values into contextual values and preferences. Monetary valuation is limited to quantifying values. Other methods are needed to understand their meaning or content, and the communal, societal and transcendental values that underpin them. Psychometric, non-analytical and interpretive methods (e.g. storytelling) can reveal those shared values. They can be combined with deliberative-analytical methods (e.g. deliberative monetary valuation and multi-criteria analysis) to provide a comprehensive valuation that can quantify values, understand their individual and shared meanings and significance, and better include ethical dimensions. Finding 7: Deliberative and social learning processes help people to understand the values held by others; they can lead to increased sharing of values and/or to greater acceptance of the decisions emerging from such processes. Deliberation clearly affects what values participants express compared to non-deliberated processes. There is also a growing body of theoretical and empirical research suggesting that deliberation has the potential to affect how people understand and shape the values of others. Although rarely considered in the economic literature, the concept of social learning helps to explain some of the processes involved in deliberation. The extent to which deliberation or social learning helps participants express and shape values will depend upon the frequency and depth of interactions and the timescale over which interactions occur. Only a shift in cultural values (e.g. less emphasis on material wealth), reflected in other societal institutions (e.g. changes in the indicators used to measure national progress) is likely to achieve sustainable outcomes in the long-term. Finding 8: Media analysis is a promising avenue for characterising different types of shared values at a large scale, as well as assessing the conflicts between the communal values of different sectors of society. There has been a marked increase in public interest in environmental issues over the last decade, which is reflected in their increased media coverage. Media content and discourse analysis is able to distinguish and characterise the plurality of cultural, societal and transcendental values and their interrelationships, and can offer a picture of the self- and other-regarding values that underpin environmental issues and conflicts. Social media can provide a further forum for understanding societal and communal values surrounding environmental issues. Finding 9: Aesthetic and spiritual values of ecosystems have a strong non-instrumental component. While they benefit human well-being, they should not simply be classified as just ‘services’ or ‘benefits’. Many spiritual discourses about nature resist talk of consequentialist benefits and economic analysis. These discourses counter assertions of the disenchantment of the world, which is associated with an instrumental environmental ethic and the commodification of nature. Allowing the possibility of enchantment can be a richer way of understanding our experience of nature and alerts us to the limitations of using economic models for valuation and informing decisions about these profound cultural ecosystem ‘services’. Faith communities have experience of using these non-utilitarian values in their own decision-making and provide models that could be adapted for use in environmental decision-making. Finding 10: Subjective well-being measures provide a useful means of assessing ‘intangible’ cultural ecosystem services and their benefits. Different user groups associate common elements of subjective well-being with environmental settings, providing opportunities for development of standardised measures. In the UK NEAFO, key facets of well-being associated with places in nature across different user groups included: engagement with nature (incorporating elements of connectedness, getting to know nature and the beauty of nature, and taking care of a place); therapeutic benefits (including physical and mental aspects of health); place identity (including a sense of place and belonging); spiritual value (in the sense of feeling connected or responsible to something larger than oneself); social bonding with others; and transformative and memorable experiences. Further empirical work with different user groups and environmental settings would allow for the continued development of a standardised tool for large-scale non-monetary assessment of cultural ecosystem services

The value of carbon sequestration and storage in coastal habitats

Coastal margin habitats are globally significant in terms of their capacity to sequester and store carbon, but their continuing decline, due to environmental change and human land use decisions, is reducing their capacity to provide this ecosystem service. In this paper the UK is used as a case study area to develop methodologies to quantify and value the ecosystem service of blue carbon sequestration and storage in coastal margin habitats. Changes in UK coastal habitat area between 1900 and 2060 are documented, the long term stocks of carbon stored by these habitats are calculated, and the capacity of these habitats to sequester CO2 is detailed. Changes in value of the carbon sequestration service of coastal habitats are then projected for 2000–2060 under two scenarios, the maintenance of the current state of the habitat and the continuation of current trends of habitat loss. If coastal habitats are maintained at their current extent, their sequestration capacity over the period 2000–2060 is valued to be in the region of £1 billion UK sterling (3.5% discount rate). However, if current trends of habitat loss continue, the capacity of the coastal habitats both to sequester and store CO2 will be significantly reduced, with a reduction in value of around £0.25 billion UK sterling (2000–2060; 3.5% discount rate). If loss-trends due to sea level rise or land reclamation worsen, this loss in value will be greater. This case study provides valuable site specific information, but also highlights global issues regarding the quantification and valuation of carbon sequestration and storage. Whilst our ability to value ecosystem services is improving, considerable uncertainty remains. If such ecosystem valuations are to be incorporated with confidence into national and global policy and legislative frameworks, it is necessary to address this uncertainty. Recommendations to achieve this are outlined