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Transient transfection of mammalian cells using plasmid DNA is a standard method to produce adeno-associated virus (AAV) vectors allowing for flexible and scalable manufacture. Typically, three plasmids are used to encode the necessary components to facilitate vector production; however, a dual-plasmid system, termed pDG, was introduced over 2 decades ago demonstrating two components could be combined resulting in comparable productivity to triple transfection. We have developed a novel dual-plasmid system, pOXB, with an alternative arrangement of sequences that results in significantly increased AAV vector productivity and percentage of full capsids packaged in comparison to the pDG dual design and triple transfection. Here, we demonstrate the reproducibility of these findings across seven recombinant AAV genomes and multiple capsid serotypes as well as the scalability of the pOXB dual-plasmid transfection at 50-L bioreactor scale. Purified drug substance showed a consistent product quality profile in line with triple-transfected vectors, except for a substantial improvement in intact genomes packaged using the pOXB dual- transfection system. Furthermore, pOXB dual- and triple-transfection-based vectors performed consistently in vivo. The pOXB dual plasmid represents an innovation in AAV manufacturing resulting in significant process gains while maintaining the flexibility of a transient transfection platform.

During childhood we begin to develop values, including valuing the natural environment (biospheric values). Although biospheric values are believed to provide the foundation for pro-environmental behavior throughout the course of one’s life, little research has investigated these values in children. The present study aimed to investigate the relationships between children’s endorsement of biospheric values, their pro-environmental behaviors, and their perception of their friends’ and peers’ endorsement of biospheric values. Moreover, we investigated whether these values and behaviors, as well as the hypothesized relationships, were affected by educational programmes that were already implemented at schools. The results showed that children generally strongly endorse biospheric values, and that biospheric values were positively related to some personal and group pro-environmental behaviors. The study also found that, as in previous research with adults, the participants believed that their friends and peers endorsed biospheric values significantly less than they themselves did. Environmental educational programs were partially effective in reducing the participants’ underestimation of their friends’ biospheric values and increased the likelihood of some group pro-environmental behaviors. Our findings highlight the need for further research to investigate the effects of group pro-environmental behaviors and the perception of group values.

Policy discourse favours the idea of “ageing in place” but many older people move home and into different kinds of residential settings. This article extends the understanding of how relocation can promote as well as diminish older people’s well-being. Using relational understandings of place and capabilities (people’s freedoms and opportunities to be and to do what they value) we explored well-being across the relocation trajectories of 21 people aged 65–91 years living in diverse residential settings in Scotland. We found that a diverse array of capabilities mattered for well-being and that relocation was often motivated by concerns to secure “at-risk” capabilities for valued activities and relationships. Moving residence impacted several other capabilities, in addition to these, both, positively and negatively. We suggest that a capability approach offers a valuable lens for understanding and supporting well-being through behavioural models of late-life relocation.

In system development, epistemic uncertainty is an ever-present possibility when reasoning about the causal factors during hazard analysis. Such uncertainty is common when complicated systems interact with one another, and it is dangerous because it impairs hazard analysis and thus increases the chance of overlooking unsafe situations. Uncertainty around causation thus needs to be managed well. Unfortunately, existing hazard analysis techniques tend to ignore unknown uncertainties, and system stakeholders rarely track known uncertainties well through the system lifecycle. In this paper, we outline an approach to managing epistemic uncertainty in existing hazard analysis techniques by focusing on known and unknown uncertainty. We have created a reference populated with a wide range of safety-critical causal relationships to recognise unknown uncertainty, and we have developed a model to systematically capture and track known uncertainty around such factors. We have also defined a process for using the reference and model to assess possible causal factors that are suspected during hazard analysis. To assess the applicability of our approach, we have analysed the widely-used MoDAF architectural model and determined that there is potential for our approach to identify additional causal factors that are not apparent from individual MoDAF views. We have also reviewed an existing safety assessment example (the ARP4761 Aircraft System analysis) and determined that our approach could indeed be incorporated into that process. We have also integrated our approach into the STPA hazard analysis technique to demonstrate its feasibility to incorporate into existing techniques. It is therefore plausible that our approach can increase safety assurance provided by hazard analysis in the face of epistemic uncertainty.

Background Digitally enabled healthcare services combine socio-technical resources to deliver the required outcomes to patients. Unintended operation of these services may result in adverse effects to the patient. Eliminating avoidable harm requires a systematic way of analysing the causal conditions, identifying opportunities for intervention. Operators of such services may be required to justify, and communicate, their safety. For example, the UK Standardisation Committee for Care Information (SCCI) standards 0129 and 0160 require a safety justification for health IT (superseded versions were known as the Information Standards Board (ISB) 0129 & 0160. Initial as well as current standards are maintained by the NHS Digital. Method A framework was designed, and applied as proof of concept, to an IT-supported clinical emergencies (A&E) service. Evaluation was done qualitatively based on the authors’ experience, identifying potential benefits of the approach. Results The applied framework encapsulates analysis, and structures the generated information, into a skeleton of an evidence-based case for safety. The framework improved management of the safety activities, assigning ownership to stakeholders (e.g. IT developer), also creating a clear and compelling safety justification. Conclusions Application of the framework significantly contributed to systematising an exploratory approach for analysing the service, in addition to existing methods such as reporting. Its application made the causal chain to harm more diaphanous. Constructing a safety case contributed to: (a) identifying potential assurance gaps, (b) planning production of information and evidence, and (c) communication of the justification by graphical unambiguous means.

This paper examines three critical environmental episodes: the development of the subsistence homestead community of Norvelt, Pennsylvania in the 1930s and 1940s, the Donora smog disaster of 1948, and the emergence of the modern environmental movement of the 1960s. It seeks to discern whether an environmental ethos, articulated so clearly by Pope Francis in Laudato Si’, has an antecedent among Catholics in northern Appalachia in the middle decades of the twentieth century.

Safe Safety arguments are key components in a safety case. Too often, safety arguments are constructed without proper reasoning. To address this, we argue that informal logic argument schemes have important roles to play in safety argument construction and reviewing process. Ten commonly used reasoning schemes in computer system safety domain are proposed. The role of informal logic dialogue games in computer system safety arguments reviewing is also discussed and the intended work in this area is proposed. It is anticipated that this work will contribute toward the development of computer system safety arguments, and help to move forward the interplay between research in informal logic and research in computer system safety engineering.