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Abstract Skilful predictions of near-term climate extremes are key to a resilient society. However, standard methods of analysing seasonal forecasts are not optimised to identify the rarer and most impactful extremes. For example, standard tercile probability maps, used in real-time regional climate outlooks, failed to convey the extreme magnitude of summer 2022 Pakistan rainfall that was, in fact, widely predicted by seasonal forecasts. Here we argue that, in this case, a strong summer La Niña provided a window of opportunity to issue a much more confident forecast for extreme rainfall than average skill estimates would suggest. We explore ways of building forecast confidence via a physical understanding of dynamical mechanisms, perturbation experiments to isolate extreme drivers, and simple empirical relationships. We highlight the need for more detailed routine monitoring of forecasts, with improved tools, to identify regional climate extremes and hence utilise windows of opportunity to issue trustworthy and actionable early warnings.

Tropilaelaps spp. are invasive mites that cause severe disease in Apis mellifera colonies. The UK has deployed an elaborate surveillance system that seeks to detect these mites early in any invasion to allow the best opportunity to eradicate any incursion. Effective field and laboratory protocols, capable of reliably detecting low numbers of mites, are key to the success of any intervention. Here we compared the efficacy of established field monitoring using brood removal with an uncapping fork, and brood 'bump' methods with novel methods for Tropilaelaps detection modified from Varroa monitoring schemes. In addition, we monitored the efficacy of the laboratory method for screening for mites in hive debris by floating mites in ethanol. Our results clearly indicated that novel methods such as uncapping infested brood with tweezers, catching mite drop using sticky traps and rolling adult bees in icing sugar were all significantly more likely to detect Tropilaelaps than existing methods such as using an uncapping fork on infested brood, or the brood 'bump' method. Existing laboratory protocols that sieved hive debris and then floated the mite containing layer failed to detect Tropilaelaps mites and new efficacious protocols were developed. Our results demonstrated that the national surveillance protocols for Tropilaelaps mite detection required modification to improve the early detection of this damaging invasive mite.

Background Stigma contributes to the negative social conditions persons with intellectual disabilities are exposed to, and it needs tackling at multiple levels. Standing Up for Myself is a psychosocial group intervention designed to enable individuals with intellectual disabilities to discuss stigmatising encounters in a safe and supportive setting and to increase their self-efficacy in managing and resisting stigma. Objectives To adapt Standing Up for Myself to make it suitable as a digital intervention; to evaluate the feasibility and acceptability of Digital Standing Up for Myself and online administration of outcome measures in a pilot; to describe usual practice in the context of the coronavirus disease 2019 pandemic to inform future evaluation. Design Adaptation work followed by a single-arm pilot of intervention delivery. Setting and participants Four third and education sector organisations. Individuals with mild-to-moderate intellectual disabilities, aged 16+, members of existing groups, with access to digital platforms. Intervention Digital Standing Up for Myself intervention. Adapted from face-to-face Standing Up for Myself intervention, delivered over four weekly sessions, plus a 1-month follow-up session. Outcomes Acceptability and feasibility of delivering Digital Standing Up for Myself and of collecting outcome and health economic measures at baseline and 3 months post baseline. Outcomes are mental well-being, self-esteem, self-efficacy in rejecting prejudice, reactions to discrimination and sense of social power. Results Adaptation to the intervention required changes to session duration, group size and number of videos; otherwise, the content remained largely the same. Guidance was aligned with digital delivery methods and a new group member booklet was produced. Twenty-two participants provided baseline data. The intervention was started by 21 participants (four groups), all of whom were retained at 3 months. Group facilitators reported delivering the intervention as feasible and suggested some refinements. Fidelity of the intervention was good, with over 90% of key components observed as implemented by facilitators. Both facilitators and group members reported the intervention to be acceptable. Group members reported subjective benefits, including increased confidence, pride and knowing how to deal with difficult situations. Digital collection of all outcome measures was feasible and acceptable, with data completeness ≥ 95% for all measures at both time points. Finally, a picture of usual practice has been developed as an intervention comparator for a future trial. Limitations The pilot sample was small. It remains unclear whether participants would be willing to be randomised to a treatment as usual arm or whether they could be retained for 12 months follow-up. Conclusions The target number of groups and participants were recruited, and retention was good. It is feasible and acceptable for group facilitators with some training and supervision to deliver Digital Standing Up for Myself. Further optimisation of the intervention is warranted. Future work To maximise the acceptability and reach of the intervention, a future trial could offer the adapted Digital Standing Up for Myself, potentially alongside the original face-to-face version of the intervention. Study registration This study was registered as ISRCTN16056848. Funding This award was funded by the National Institute for Health and Care Research (NIHR) Public Health Research programme (NIHR award ref: 17/149/03) and is published in full in Public Health Research; Vol. 12, No. 1. See the NIHR Funding and Awards website for further award information. Plain language summary People with intellectual disabilities (or ‘learning disabilities’ in United Kingdom language) are more likely to experience poor physical and mental health than the general population. Stigma (negative stereotypes, prejudice and discrimination) has been linked to lower self-esteem, quality of life, and mental and physical ill health. Efforts to empower people with intellectual disabilities themselves to challenge stigma with a view to improving well-being, health and self-esteem are lacking. In 2017, we developed Standing Up for Myself, a brief group-based programme for people with mild-to-moderate intellectual disabilities aged 16+ to address this gap. As this study got underway, face-to-face meetings were suspended due to the coronavirus disease 2019 pandemic. We used the opportunity to assess whether Standing Up for Myself could be delivered through web-based meetings. We adapted Standing Up for Myself for digital delivery, with close input from advisors with intellectual disabilities and experienced group facilitators. We then tested the digital version in charity and education settings to evaluate if Digital Standing Up for Myself could be delivered as planned and how acceptable it was to group facilitators and participants. Four groups, with a total of 22 members, signed up to try Digital Standing Up for Myself. One participant dropped out before starting Standing Up for Myself, and the other 21 continued until the end of the programme. Retention and attendance were good; participants on average attended four of the five sessions. Ninety per cent of the core programme requirements were fully delivered as detailed in the Digital Standing Up for Myself manual. Problems with technology were manageable, although facilitators found using the Standing Up for Myself Wiki platform (an online platform for storage and sharing of resources) difficult, particularly when sharing video content. Facilitators felt acceptable levels of privacy were achieved and there were no reports of undue distress. All facilitators and many group members said they would recommend Digital Standing Up for Myself to others. Group members shared how the programme benefitted them, noting increased awareness about disabilities, and for some increased confidence, pride and independence. Some had learnt how to stand up for themselves and manage difficult situations and took pride in this. Completing outcome and health cost measures via web-based meetings was acceptable and data were largely fully complete and useable. Scientific summary Background Approximately 1.4–2% of the UK population have an intellectual disability. Individuals with intellectual disabilities face substantial social and health inequalities and are at increased risk of experiencing mental health problems. One factor compounding these inequalities is the impact of stigma, whereby individuals experience negative stereotyping, prejudice and discrimination associated with intellectual disability. Interventions that seek to reduce stigma are needed at multiple levels. The Standing Up for Myself (STORM) programme targets the person with an intellectual disability themselves and seeks to empower them as agent of positive change. It seeks to empower individuals with intellectual disabilities to challenge stigma they face in everyday encounters, thus potentially improving well-being and reducing inequalities, alongside interventions at community and institutional/societal levels to reduce stigma. An earlier pilot of the STORM programme showed it to be a promising intervention which could be delivered in community, third sector (i.e. ‘charities’ that typically provide services to achieve social goals) and education settings to existing groups of individuals with intellectual disabilities, aged 16 years and above. Some issues were highlighted which were to be addressed as part of a feasibility study with the intention of progressing to a fully powered randomised controlled trial (RCT). The start of the feasibility study coincided with the outbreak of the coronavirus disease 2019 (COVID-19) pandemic and the first national lockdown during month 6 of the original project. Following a necessary pause, the study management team saw a unique opportunity to revise the study to examine the potential of STORM as a digital intervention. Adapting STORM to make it suitable for digital delivery would allow the programme to be available to a much wider audience and potentially future-proof it in the context of the ongoing pandemic. Extensive patient and public involvement (PPI) work generated evidence in line with the National Institute for Health and Care Excellence (Evidence Standards Framework for Digital Health Technologies, 2019, www.nice.org.uk/Media/Default/About/what-we-do/our-programmes/evidence-standards-framework/digital-evidence-standards-framework.pdf) framework for digital health technologies and indicated that the creation of a digital version of STORM was important to potential future users and professionals. We, therefore, proposed to adapt the STORM programme for digital delivery to groups of individuals with mild-to-moderate intellectual disabilities. Working closely with PPI partners we planned, following adaptation of the intervention, to deliver the digital programme to four groups as part of a small pilot study. We also engaged experts on digital inclusion and learning design to ensure that the adapted digital version of STORM was optimised for engagement of people with intellectual disabilities and delivery by group facilitators with varying experience and skills in digital delivery. Objectives to adapt the existing STORM intervention for online delivery (Digital STORM), ensuring the content, number of sessions and direct contact time were the same for both STORM and Digital STORM; to pilot the Digital STORM intervention in order to investigate the feasibility of recruitment to and retention of participants in Digital STORM; and adherence, fidelity and acceptability of Digital STORM, when delivered to groups of people with mild-to-moderate intellectual disabilities online; to test digital administration of the study outcome and health economics measures; to build on community assessments to describe what usual practice might look like for groups of people with mild-to-moderate intellectual disabilities in the wake of COVID-19, to inform a potential future trial. Methods Design and procedure Digital STORM was an adaptation of the original STORM intervention, designed for online delivery, piloted with four groups of young people and adults with mild-to-moderate intellectual disabilities. Adaptation and pilot work took place during months 14–21 of the overall 24-month project in distinct phases of work: Intervention adaptation (months 14–17) An Intervention Adaptation Group (IAG) was established to oversee the adaptation and report progress to oversight committees. The group included all members of the PPI advisory group (people with intellectual disabilities, the independent co-chair), experienced group facilitators from third and education sector organisations (from our stakeholder group), Mencap as our intervention delivery partner, digital inclusion experts and members of the research team. The focus of this group was to maximise access to and engagement with Digital STORM to ensure it would be inclusive and to address potential barriers to access to and/or engagement with the intervention. Pilot of Digital STORM (months 18–21) The adapted intervention was then piloted with four groups (N = 22). Priority was given to groups that had expressed interest in participating in STORM at the point of having to pause the original study in March 2020 due to the first national coronavirus disease 2019 (COVID-19) lockdown. Of the four pilot groups, at least one would need to do some additional work to allow all its group members to access Digital STORM. This would allow us to explore ‘live’ how issues relating to access to technology, support, and provisions to ensure privacy are managed. Following delivery of the adapted digital intervention, pilot group facilitators were interviewed about their experiences to assess barriers and facilitators to implementation. Participants took part in focus groups to access their views on the intervention, the delivery mechanisms, and the intervention’s subjective impact. The focus groups were co-led by a member of the PPI advisory group and a researcher in three cases and by a researcher alone in one case and recorded. Decision phase (month 22) Oversight committees met at regular intervals throughout the adaptation and pilot phases to monitor progress. They reviewed findings from the pilot against the progression criteria and made recommendations to the National Institute for Health and Care Research (NIHR) regarding the potential for a future funding application. Inclusion and exclusion criteria Groups from third and education sector organisations were included where they had a willing facilitator and organisational support in place, were meeting or restarting meetings as a group for at least 3 further months and were willing to replace five of their meetings with Digital STORM. Groups needed at least three and no more than eight members with intellectual disabilities to participate. Groups were excluded if they were run as part of the National Health Service, or if some of their regular members declined taking part in Digital STORM and if it was not possible to find alternative meeting times for those who wanted to participate. Individual participants were included if they were aged 16 years or older, had mild-to-moderate intellectual disabilities, capacity to provide informed consent, were able to complete the outcome measures and could engage with the intervention in English. They needed to be a member of an established (educational, activity, social or self-advocacy focused) group; to have access to the internet, a device to join web meetings, and support to access web-based meetings when needed. Participants were excluded from the research if they did not provide consent. Intervention Adaptation of the STORM intervention for delivery using web-based video meetings (Digital STORM), consisting of four weekly 90-minute sessions and a 90-minute follow-up session (delivered around 4 weeks after session four). An intervention manual and a Wiki (a web platform designed as both a repository of intervention resources and an aide to delivering session content) was provided to facilitators. Outcomes The primary outcome of the adaptation phase was the feasibility and acceptability of delivering STORM to groups in a web-delivered format (Digital STORM). Secondary outcomes of the adaptation phase were the feasibility of remotely collecting outcomes, health economics and process data and the description of ‘Usual practice’, that is, activities usually undertaken within the group setting, in a changed service delivery context. Analyses Pilot data (baseline demographics, responses to outcome measures, adherence and fidelity ratings) were described descriptively. The feasibility of economic evaluation was assessed using completion rates of included measures [Service Information Schedule (SIS), EuroQol-Youth version, and Client Service Receipt Inventory]. Intervention costs were calculated using SIS data. Qualitative interviews and focus groups were conducted to explore barriers and facilitators to participation in the intervention. Interview and focus group transcripts were analysed thematically using Framework analysis; a sample of transcripts was double coded. Progression criteria were included to determine suitability for moving to a future trial of Digital STORM. Results The STORM intervention was successfully adapted for online delivery (objective 1). The IAG addressed issues such as access to the digital intervention, engagement with it and potential risks to managing participant privacy. Minor adjustments were made to the STORM intervention to allow for digital delivery. Content was slightly streamlined to ensure each session could be covered within the 60-minute sessions. Resources were created to support participation in online group work (for participants) and for running sessions (for facilitators). A revised intervention logic model for Digital STORM was also created. Digital STORM was both feasible and acceptable when delivered to groups online (objective 2). Targets for recruitment into the pilot of Digital STORM were met with 4 groups taking part and 22 participants providing informed consent, 1 of whom dropped out before starting the intervention. There was good attendance across sessions (median attendance 5 out of 5 sessions, with 20 of 21 participants attending 3 or more sessions) and the majority of participants missed no more than 15 minutes of any one intervention session due to technical difficulties. Accordingly, there was a strong indicator of feasibility for progression to a future trial. There was also a strong indicator for the acceptability of Digital STORM. Facilitators found the recording of sessions acceptable and over 90% of the core intervention requirements were met in full. Feasibility and acceptability were also examined through interviews (with facilitators) and focus groups (with participants). Resources supported facilitators’ delivery of sessions though some reported challenges when playing and sharing videos in online meetings and with trying to co-ordinate moving between video sharing via the STORM Wiki platform, the manual and operating the meeting platform. Feasibility of administering study outcome measures was also demonstrated (objective 3). Participants completed outcome measures via web-based platforms at baseline and post intervention. This enabled the research team to recruit from a wider geographical area than would have been possible using face-to-face data collection methods. Data completeness was very high – only one response was missing across all measures. Similarly, data completeness for economic evaluation was very good with no barriers to future data collection identified. Group members’ experience of completing measures was positive and they felt positive about the digital approach as a method for data collection in future. Finally, it was determined that many organisations delivering group sessions had made the transition to online delivery and were running virtual groups. This supports the possibility of including a control arm that described ‘usual practice’ as part of a future trial (objective 4). Conclusions The STORM digital adaptation and pilot was a well-delivered package of work. Results evidence that all progression criteria were achieved in full. Oversight committees therefore recommend progression to a full trial. Strengths and limitations The key strength of this work was the ability of the team to work flexibly and creatively to adapt to the changing situation of the pandemic. Involvement of experts by experience meant the rationale for creating a digital version of STORM was supported by all stakeholders and the accessibility of the processes and resources was considered carefully from a user perspective. A number of limitations need noting. These include the small sample size for the pilot; long-term retention was not established; those taking part all did so with the knowledge they would receive the intervention, thus the ability to randomise was not confirmed; the video sharing platform was not optimal – other approaches might have worked better; qualitative work could not capture the views of all those who took part in the pilot. Study registration This study was registered as ISRCTN16056848. Funding This award was funded by the National Institute for Health and Care Research (NIHR) Public Health Research programme (NIHR award ref: 17/149/03) and is published in full in Public Health Research; Vol. 12, No. 1. See the NIHR Funding and Awards website for further award information.

Recycled aggregate (RA) from construction and demolition waste can readily be used to replace natural aggregate in concrete. Nonetheless, the poor quality of RA adversely affects the properties of recycled aggregate concrete, limiting its use to only non-structural applications. This study examined the effects of various treatment methods by testing the aggregate impact value (AIV) and water absorption (WA) of recycled coarse aggregate before and after treatments as an indicator to examine the efficiency of these treatments. The results showed that the untreated RA achieved 17% and 6.1% for AIV and WA, respectively. Accelerated carbonation treatment at 50% CO2 concentration level for six days of CO2 exposure time achieved the best results among other carbonation conditions, showing 11% and 46% enhancements in the AIV and WA, respectively. The RA treated with cyclic limewater combined with accelerated carbonation exhibited better improvement in the AIV and WA compared to the sole use of accelerated carbonation treatment. Coating RA with cement-silica fume slurry at 0.1 mm coating thickness was found to be optimal, achieving 12% and 54% enhancements in the AIV and WA, respectively. Soaking RA in a 10% Portland cement - pulverized fuel ash - silica fume solution for 4 h was found to be optimal, recording enhancements of 13% and 54% in the AIV and the WA, respectively among other soaking solutions. Soaking RA in 5% sodium silicate – silica fume solution for 4 h was found to be optimal, obtaining 8% and 33% enhancements in the AIV and the WA, respectively. The treatment techniques proposed can be a powerful tool for promoting the use of RA in the construction industry.

English Language Teaching (ELT) in Mexico covers a wide range of situations and results, from bilingual schools achieving excellent results, through language centres achieving generally good results, to common core English programmes in universities that start students at beginner level for the third or fourth time, after a decade or more of English classes at school, and generally fail to get most students beyond A2 level. Those common core university English programmes and other evidence suggest that most ELT in Mexico, especially school ELT, is failing to achieve worthwhile results. Sadly, we know too little for sure about both the successes and the failures of ELT in Mexico, though we can assume or guess a lot. This article considers what we really know about ELT in Mexico, what we do not know and urgently need to find out in order to radically improve the general situation, and how we might find it out.

The recent emergence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the underlying cause of Coronavirus Disease 2019 (COVID-19), has led to a worldwide pandemic causing substantial morbidity, mortality, and economic devastation. In response, many laboratories have redirected attention to SARS-CoV-2, meaning there is an urgent need for tools that can be used in laboratories unaccustomed to working with coronaviruses. Here we report a range of tools for SARS-CoV-2 research. First, we describe a facile single plasmid SARS-CoV-2 reverse genetics system that is simple to genetically manipulate and can be used to rescue infectious virus through transient transfection (without in vitro transcription or additional expression plasmids). The rescue system is accompanied by our panel of SARS-CoV-2 antibodies (against nearly every viral protein), SARS-CoV-2 clinical isolates, and SARS-CoV-2 permissive cell lines, which are all openly available to the scientific community. Using these tools, we demonstrate here that the controversial ORF10 protein is expressed in infected cells. Furthermore, we show that the promising repurposed antiviral activity of apilimod is dependent on TMPRSS2 expression. Altogether, our SARS-CoV-2 toolkit, which can be directly accessed via our website at https://mrcppu-covid.bio/, constitutes a resource with considerable potential to advance COVID-19 vaccine design, drug testing, and discovery science.

Of the 16 non-structural proteins (Nsps) encoded by SARS CoV-2, Nsp3 is the largest and plays important roles in the viral life cycle. Being a large, multidomain, transmembrane protein, Nsp3 has been the most challenging Nsp to characterize. Encoded within Nsp3 is the papain-like protease domain (PLpro) that cleaves not only the viral polypeptide but also K48-linked polyubiquitin and the ubiquitin-like modifier, ISG15, from host cell proteins. We here compare the interactors of PLpro and Nsp3 and find a largely overlapping interactome. Intriguingly, we find that near full length Nsp3 is a more active protease compared to the minimal catalytic domain of PLpro. Using a MALDI-TOF based assay, we screen 1971 approved clinical compounds and identify five compounds that inhibit PLpro with IC50s in the low micromolar range but showed cross reactivity with other human deubiquitinases and had no significant antiviral activity in cellular SARS-CoV-2 infection assays. We therefore looked for alternative methods to block PLpro activity and engineered competitive nanobodies that bind to PLpro at the substrate binding site with nanomolar affinity thus inhibiting the enzyme. Our work highlights the importance of studying Nsp3 and provides tools and valuable insights to investigate Nsp3 biology during the viral infection cycle.

The inhibitory synaptic protein collybistin (CB) and GABAAR-α subunits are thought to interact, but strength and specificity are unclear. Here the authors study the CB–α2 interaction and show that a mouse mutated in the CB-binding region of α2 displays a loss of specific synapses and seizure.

Theorisations of comics and graphic narrative have tended to focus on readership rather than creation. Even McCloud’s seminal Understanding Comics (1993) focuses on interpretation, though the later Making Comics (2006) pays more attention to the work of the creator. McCloud incorporates into his theory five ‘choices of the comic creator’: choices of Moment, Frame, Image, Word and Flow.However, these may not be the most salient choices for a creator to take; nor do they necessarily occur in that order. Using these alongside McCloud’s other notions (notably ‘transitions’) with a group of sixth form comics creators led me to reconsider approaches to comics creation, as well as reading, in my PhD research ‘Making Meanings with Comics: A Functional Approach to Graphic Narrative’. In this work, I take models of meaning-making from the functional linguistics of M.A.K. Halliday, and adapt that framework to specify how comics make meaning. As a linguistics-inspired approach, this takes seriously the production of meaning as well as the reception.In this graphic article, I give a brief illustrated overview of this view of comics meaning making, and present a set of new ‘choices for the comics creator’ which arose from it. These new choices include: Choice of Character Design, Choice of ‘Verb Style’, Choice of Framing, Choice of Density, Choice of Metonymy. I offer a critique of McCloud’s choices, and account for the proposed alternatives, indicating the theory that led to them with illustrative examples, drawn in the main from my own comics created in that sixth form workshop alongside students. The article is drawn digitally and incorporates extracts of comics pages as well as diagrams to illustrate the approach to comics that informs the five ‘choices’ described.

A Theory of Narrative Drawing, by Simon Grennan, Palgrave Macmillan; XII, 277 pages, 16 b/w illustrations, 10 illustrations in colour; eBook ISBN 978-1-137-51844-6; DOI: https://doi.org/10.1057/978-1-137-51844-6; Hardcover ISBN 978-1-137-52165-1 This review in graphic form responds to Simon Grennan’s 2017 monograph ‘A Theory of Narrative Drawing’, describing its form and key ideas, and commenting on its contribution to the field, whilst enacting the graphic mark-making which is its focus. The review concludes that though some more framing material would have been useful for readers, the book repays a second reading and makes an invaluable contribution to the view of comics as an active process of meaning-making.

Cancer is sometimes depicted as a reversion to single cell behavior in cells adapted to live in a multicellular assembly. If this is the case, one would expect that mutation in cancer disrupts functional mechanisms that suppress cell-level traits detrimental to multicellularity. Such mechanisms should have evolved with or after the emergence of multicellularity. This leads to two related, but distinct hypotheses: 1) Somatic mutations in cancer will occur in genes that are younger than the emergence of multicellularity (1000 million years [MY]); and 2) genes that are frequently mutated in cancer and whose mutations are functionally important for the emergence of the cancer phenotype evolved within the past 1000 million years, and thus would exhibit an age distribution that is skewed to younger genes. In order to investigate these hypotheses we estimated the evolutionary ages of all human genes and then studied the probability of mutation and their biological function in relation to their age and genomic location for both normal germline and cancer contexts. We observed that under a model of uniform random mutation across the genome, controlled for gene size, genes less than 500 MY were more frequently mutated in both cases. Paradoxically, causal genes, defined in the COSMIC Cancer Gene Census, were depleted in this age group. When we used functional enrichment analysis to explain this unexpected result we discovered that COSMIC genes with recessive disease phenotypes were enriched for DNA repair and cell cycle control. The non-mutated genes in these pathways are orthologous to those underlying stress-induced mutation in bacteria, which results in the clustering of single nucleotide variations. COSMIC genes were less common in regions where the probability of observing mutational clusters is high, although they are approximately 2-fold more likely to harbor mutational clusters compared to other human genes. Our results suggest this ancient mutational response to stress that evolved among prokaryotes was co-opted to maintain diversity in the germline and immune system, while the original phenotype is restored in cancer. Reversion to a stress-induced mutational response is a hallmark of cancer that allows for effectively searching "protected" genome space where genes causally implicated in cancer are located and underlies the high adaptive potential and concomitant therapeutic resistance that is characteristic of cancer.

Mutations in Park8, encoding for the multidomain Leucine-rich repeat kinase 2 (LRRK2) protein, comprise the predominant genetic cause of Parkinson's disease (PD). G2019S, the most common amino acid substitution activates the kinase two- to threefold. This has motivated the development of LRRK2 kinase inhibitors; however, poor consensus on physiological LRRK2 substrates has hampered clinical development of such therapeutics. We employ a combination of phosphoproteomics, genetics, and pharmacology to unambiguously identify a subset of Rab GTPases as key LRRK2 substrates. LRRK2 directly phosphorylates these both in vivo and in vitro on an evolutionary conserved residue in the switch II domain. Pathogenic LRRK2 variants mapping to different functional domains increase phosphorylation of Rabs and this strongly decreases their affinity to regulatory proteins including Rab GDP dissociation inhibitors (GDIs). Our findings uncover a key class of bona-fide LRRK2 substrates and a novel regulatory mechanism of Rabs that connects them to PD.

This paper provides a discussion of the methodological approach to a research project that builds on previous studies of effective argumentation pedagogy undertaken by one of the authors. In this study, teachers from six schools in a city location are taking part in a one-year project to use new web-based professional development materials to advance their practice in using group discussion and argumentation in science. The teachers attend workshops on planning and organising effective group-work, introducing argument and sustaining small group discussion, and curriculum planning to promote argumentation activity. Between sessions the teachers develop each aspect of pedagogy in their schools. Data are collected from each of the participating schools using teacher survey and interviews to ascertain perspectives on argumentation practice in science, pedagogical strategies used, changes in practice and collaborative working with colleagues in school. The paper focuses on the issues of research design in determining the impact of the workshops.Este artigo traz uma discussão sobre a abordagem metodológica de um projeto de pesquisa que se baseia em estudos anteriores sobre a pedagogia da argumentação eficaz, realizado por um dos autores. Neste estudo, professores de seis escolas estão participando de um projeto, com duração de um ano, para usar novos materiais de desenvolvimento profissionais baseados na internet para a melhoria da sua prática no uso de grupo de discussão e argumentação em ciências. Os professores participam de oficinas sobre planejamento e organização de grupos de trabalho eficientes, introduzindo argumentação e mantendo a discussão em pequenos grupos, e do planejamento de currículos para promover a atividade argumentativa. Entre as sessões, os professores desenvolvem cada aspecto da pedagogia em suas escolas. Os dados são levantados em cada uma das escolas participantes por meio de questionários e entrevistas junto aos professores para averiguar as perspectivas sobre a prática da argumentação em ciência, estratégias pedagógicas utilizadas, mudanças nas práticas e trabalho colaborativo com os colegas na escola. O artigo enfoca questões sobre o delineamento da pesquisa para determinar o impacto das oficinas.

Narrative Structure in Comics builds on Postema’s PhD thesis to present for a more general audience her focus on the ‘gap’ in comics and its place in the process of reading graphic narrative, from the detailed textual level up to the level of narrative structure overall. Postema's readings of comics texts are well-argued and illuminating; the breadth of theory brought together here, and the range of exemplars used in analysis, make Narrative Structure in Comics an invaluable reader for those interested in engaging with the practical application of medium-specific theory to comics texts themselves.

Empirical studies give increased support for the hypothesis that the sporadic form of cancer is an age-related metabolic disease characterized by: (a) metabolic dysregulation with random abnormalities in mitochondrial DNA, and (b) metabolic alteration – the compensatory upregulation of glycolysis to offset mitochondrial impairments. This paper appeals to the theory of Quantum Metabolism and the principles of natural selection to formulate a conceptual framework for a quantitative analysis of the origin and proliferation of the disease. Quantum Metabolism, an analytical theory of energy transduction in cells inspired by the methodology of the quantum theory of solids, elucidates the molecular basis for differences in metabolic rate between normal cells, utilizing predominantly oxidative phosphorylation, and cancer cells utilizing predominantly glycolysis. The principles of natural selection account for the outcome of competition between the two classes of cells. Quantum Metabolism and the principles of natural selection give an ontogenic and evolutionary rationale for cancer proliferation and furnish a framework for effective therapeutic strategies to impede the spread of the disease.

α-synuclein (αS) is a cellular protein mostly known for the association of its aggregated forms with a variety of diseases that include Parkinson's disease and Dementia with Lewy Bodies. While the role of αS in disease is well documented there is currently no agreement on the physiological function of the normal isoform of the protein. Here we provide strong evidence that αS is a cellular ferrireductase, responsible for reducing iron (III) to bio available iron (II). The recombinant form of the protein has a V(Max) of 2.72 nmols/min/mg and K(m) 23 µM. This activity is also evident in lysates from neuronal cell lines overexpressing αS. This activity is dependent on copper bound to αS as a cofactor and NADH as an electron donor. Overexpression of α-synuclein by cells significantly increases the percentage of iron (II) in cells. The common disease mutations associated with increased susceptibility to PD show no [corrected] differences in activity or iron (II) levels. This discovery may well provide new therapeutic targets for PD and Lewy body dementias.

Prion diseases are considered to be transmissible. The existence of sporadic forms of prion diseases such as scrapie implies an environmental source for the infectious agent. This would suggest that under certain conditions the prion protein, the accepted agent of transmission, can survive in the environment. We have developed a novel technique to extract the prion protein from soil matrices. Previous studies have suggested that environmental manganese is a possible risk factor for prion diseases. We have shown that exposure to manganese is a soil matrix causes a dramatic increase in prion protein survival (approximately 10 fold) over a two year period. We have also shown that manganese increases infectivity of mouse passaged scrapie to culture cells by 2 logs. These results clearly verify that manganese is a risk factor for both the survival of the infectious agent in the environment and its transmissibility.

Minimal residual disease detection, used for clinical management of children with acute lymphoblastic leukemia, can be performed by molecular analysis of antigen-receptor gene rearrangements or by flow cytometric analysis of aberrant immunophenotypes. For flow minimal residual disease to be incorporated into larger national and international trials, a quality assured, standardized method is needed which can be performed in a multi-center setting. We report a four color, flow cytometric protocol established and validated by the UK acute lymphoblastic leukemia Flow minimal residual disease group. Quality assurance testing gave high inter-laboratory agreement with no values differing from a median consensus value by more than one point on a logarithmic scale. Prospective screening of B-ALL patients (n=206) showed the method was applicable to 88.3% of patients. The minimal residual disease in bone marrow aspirates was quantified and compared to molecular data. The combined risk category concordance (minimal residual disease levels above or below 0.01%) was 86% (n=134). Thus, this standardized protocol is highly reproducible between laboratories, sensitive, applicable, and shows good concordance with molecular-based analysis.

Insertional mutations leading to expansion of the octarepeat domain of the prion protein (PrP) are directly linked to prion disease. While normal PrP has four PHGGGWGQ octapeptide segments in its flexible N-terminal domain, expanded forms may have up to nine additional octapeptide inserts. The type of prion disease segregates with the degree of expansion. With up to four extra octarepeats, the average onset age is above 60 years, whereas five to nine extra octarepeats results in an average onset age between 30 and 40 years, a difference of almost three decades. In wild-type PrP, the octarepeat domain takes up copper (Cu(2+)) and is considered essential for in vivo function. Work from our lab demonstrates that the copper coordination mode depends on the precise ratio of Cu(2+) to protein. At low Cu(2+) levels, coordination involves histidine side chains from adjacent octarepeats, whereas at high levels each repeat takes up a single copper ion through interactions with the histidine side chain and neighboring backbone amides. Here we use both octarepeat constructs and recombinant PrP to examine how copper coordination modes are influenced by octarepeat expansion. We find that there is little change in affinity or coordination mode populations for octarepeat domains with up to seven segments (three inserts). However, domains with eight or nine total repeats (four or five inserts) become energetically arrested in the multi-histidine coordination mode, as dictated by higher copper uptake capacity and also by increased binding affinity. We next pooled all published cases of human prion disease resulting from octarepeat expansion and find remarkable agreement between the sudden length-dependent change in copper coordination and onset age. Together, these findings suggest that either loss of PrP copper-dependent function or loss of copper-mediated protection against PrP polymerization makes a significant contribution to early onset prion disease.