Cross-regulation between Aurora B and Citron kinase controls midbody architecture in cytokinesis.

Cytokinesis culminates in the final separation, or abscission, of the two daughter cells at the end of cell division. Abscission relies on an organelle, the midbody, which forms at the intercellular bridge and is composed of various proteins arranged in a precise stereotypic pattern. The molecular mechanisms controlling midbody organization and function, however, are obscure. Here we show that proper midbody architecture requires cross-regulation between two cell division kinases, Citron kinase (CIT-K) and Aurora B, the kinase component of the chromosomal passenger complex (CPC). CIT-K interacts directly with three CPC components and is required for proper midbody architecture and the orderly arrangement of midbody proteins, including the CPC. In addition, we show that CIT-K promotes Aurora B activity through phosphorylation of the INCENP CPC subunit at the TSS motif. In turn, Aurora B controls CIT-K localization and association with its central spindle partners through phosphorylation of CIT-K's coiled coil domain. Our results identify, for the first time, a cross-regulatory mechanism between two kinases during cytokinesis, which is crucial for establishing the stereotyped organization of midbody proteins.C.M. holds a Cambridge Cancer Centre PhD studentship, which is supported by Cancer Research UK and the MRC. Z.I.B. was supported by a Gwynaeth Pretty PhD studentship and the BlueGnome Molecular Genetics Fund from the Department of Pathology, University of Cambridge, and was also a recipient of a short-term EMBO fellowship and a fellowship from the Cambridge Philosophical Society. G.C. and M.G. are supported by a grant ‘Progetto di Ricerca di Interesse Nazionale’ (PRIN 2012) from the Ministero dell'Istruzione, Università e Ricerca. P.P.D. is the recipient of the Maplethorpe Fellowship from Murray Edwards College, Cambridge, UK.This is the final version of the article. It first appeared from Royal Society Publishing via https://doi.org/10.1098/rsob.16001

Novel Fas-TNFR chimeras that prevent Fas ligand-mediated kill and signal synergistically to enhance CAR T cell efficacy

The hostile tumor microenvironment limits the efficacy of adoptive cell therapies. Activation of the Fas death receptor initiates apoptosis and disrupting these receptors could be key to increasing CAR T cell efficacy. We screened a library of Fas-TNFR proteins identifying several novel chimeras that not only prevented Fas ligand-mediated kill, but also enhanced CAR T cell efficacy by signaling synergistically with the CAR. Upon binding Fas ligand, Fas-CD40 activated the NF-κB pathway, inducing greatest proliferation and IFN-γ release out of all Fas-TNFRs tested. Fas-CD40 induced profound transcriptional modifications, particularly genes relating to the cell cycle, metabolism, and chemokine signaling. Co-expression of Fas-CD40 with either 4-1BB- or CD28-containing CARs increased in vitro efficacy by augmenting CAR T cell proliferation and cancer target cytotoxicity, and enhanced tumor killing and overall mouse survival in vivo. Functional activity of the Fas-TNFRs were dependent on the co-stimulatory domain within the CAR, highlighting crosstalk between signaling pathways. Furthermore, we show that a major source for Fas-TNFR activation derives from CAR T cells themselves via activation-induced Fas ligand upregulation, highlighting a universal role of Fas-TNFRs in augmenting CAR T cell responses. We have identified Fas-CD40 as the optimal chimera for overcoming Fas ligand-mediated kill and enhancing CAR T cell efficacy

Exploration of T cell immune responses by expression of a dominant-negative SHP1 and SHP2

SHP1 and SHP2 are SH2 domain-containing proteins which have inhibitory phosphatase activity when recruited to phosphorylated ITIMs and ITSMs on inhibitory immune receptors. Consequently, SHP1 and SHP2 are key proteins in the transmission of inhibitory signals within T cells, constituting an important point of convergence for diverse inhibitory receptors. Therefore, SHP1 and SHP2 inhibition may represent a strategy for preventing immunosuppression of T cells mediated by cancers hence improving immunotherapies directed against these malignancies. Both SHP1 and SHP2 contain dual SH2 domains responsible for localization to the endodomain of inhibitory receptors and a protein tyrosine phosphatase domain which dephosphorylates and thus inhibits key mediators of T cell activation. We explored the interaction of the isolated SH2 domains of SHP1 and SHP2 to inhibitory motifs from PD1 and identified strong binding of both SH2 domains from SHP2 and more moderate binding in the case of SHP1. We next explored whether a truncated form of SHP1/2 comprising only of SH2 domains (dSHP1/2) could act in a dominant negative fashion by preventing docking of the wild type proteins. When co-expressed with CARs we found that dSHP2 but not dSHP1 could alleviate immunosuppression mediated by PD1. We next explored the capacity of dSHP2 to bind with other inhibitory receptors and observed several potential interactions. In vivo we observed that the expression of PDL1 on tumor cells impaired the ability of CAR T cells to mediate tumor rejection and this effect was partially reversed by the co-expression of dSHP2 albeit at the cost of reduced CAR T cell proliferation. Modulation of SHP1 and SHP2 activity in engineered T cells through the expression of these truncated variants may enhance T cell activity and hence efficacy in the context of cancer immunotherapy

Exploration of T cell immune responses by expression of a dominant-negative SHP1 and SHP2

SHP1 and SHP2 are SH2 domain-containing proteins which have inhibitory phosphatase activity when recruited to phosphorylated ITIMs and ITSMs on inhibitory immune receptors. Consequently, SHP1 and SHP2 are key proteins in the transmission of inhibitory signals within T cells, constituting an important point of convergence for diverse inhibitory receptors. Therefore, SHP1 and SHP2 inhibition may represent a strategy for preventing immunosuppression of T cells mediated by cancers hence improving immunotherapies directed against these malignancies. Both SHP1 and SHP2 contain dual SH2 domains responsible for localization to the endodomain of inhibitory receptors and a protein tyrosine phosphatase domain which dephosphorylates and thus inhibits key mediators of T cell activation. We explored the interaction of the isolated SH2 domains of SHP1 and SHP2 to inhibitory motifs from PD1 and identified strong binding of both SH2 domains from SHP2 and more moderate binding in the case of SHP1. We next explored whether a truncated form of SHP1/2 comprising only of SH2 domains (dSHP1/2) could act in a dominant negative fashion by preventing docking of the wild type proteins. When co-expressed with CARs we found that dSHP2 but not dSHP1 could alleviate immunosuppression mediated by PD1. We next explored the capacity of dSHP2 to bind with other inhibitory receptors and observed several potential interactions. In vivo we observed that the expression of PDL1 on tumor cells impaired the ability of CAR T cells to mediate tumor rejection and this effect was partially reversed by the co-expression of dSHP2 albeit at the cost of reduced CAR T cell proliferation. Modulation of SHP1 and SHP2 activity in engineered T cells through the expression of these truncated variants may enhance T cell activity and hence efficacy in the context of cancer immunotherapy

Characterising illness stages and recovery trajectories of eating disorders in young people via remote measurement technology (STORY):A multi-centre prospective cohort study protocol

Background: Eating disorders (EDs) are serious, often chronic, conditions associated with pronounced morbidity, mortality, and dysfunction increasingly affecting young people worldwide. Illness progression, stages and recovery trajectories of EDs are still poorly characterised. The STORY study dynamically and longitudinally assesses young people with different EDs (restricting; bingeing/bulimic presentations) and illness durations (earlier; later stages) compared to healthy controls. Remote measurement technology (RMT) with active and passive sensing is used to advance understanding of the heterogeneity of earlier and more progressed clinical presentations and predictors of recovery or relapse. Methods: STORY follows 720 young people aged 16–25 with EDs and 120 healthy controls for 12 months. Online self-report questionnaires regularly assess ED symptoms, psychiatric comorbidities, quality of life, and socioeconomic environment. Additional ongoing monitoring using multi-parametric RMT via smartphones and wearable smart rings (‘Ōura ring’) unobtrusively measures individuals’ daily behaviour and physiology (e.g., Bluetooth connections, sleep, autonomic arousal). A subgroup of participants completes additional in-person cognitive and neuroimaging assessments at study-baseline and after 12 months. Discussion: By leveraging these large-scale longitudinal data from participants across ED diagnoses and illness durations, the STORY study seeks to elucidate potential biopsychosocial predictors of outcome, their interplay with developmental and socioemotional changes, and barriers and facilitators of recovery. STORY holds the promise of providing actionable findings that can be translated into clinical practice by informing the development of both early intervention and personalised treatment that is tailored to illness stage and individual circumstances, ultimately disrupting the long-term burden of EDs on individuals and their families.</p

Future Experiences: Sustainable Development and the Global South

The Sustainable Development and the Global South project was jointly conceived by the Innovation School at Glasgow School of Art in partnership with the Sustainable Futures in Africa Network (SFA), and the University of Glasgow. Graduating final year BDes Product Design students from the Innovation School were presented with a challenge-based project to produce a vision of the future based on current trends that relate to Sustainable Development work and the Global South. This project involved working closely with researchers, academics and professionals specialising in human geography, education, health, environment, engineering, cultural practice and community engagement who are part of the Sustainable Futures in Africa Network which includes a Scottish hub, led from the University of Glasgow. Included in the network was a representative from an NGO that builds schools in Malawi, an entrepreneur who runs an ethical clothing company that partners with producers in the Global South, a senior governance officer from the UK Government’s Department for International Development (DFID), a research network administrator, and international graduate students from Africa based at Scottish institutions. In addition to the SFA, external experts from design studio AndThen and GOODD design consultancy were engaged. The objective of this project was to investigate, in both analytical and speculative ways, future forms and functions of Sustainable Development work in relation to the Global South in ten years from now, to develop future scenarios and design the artefacts, services and the experiences associated with these future visions. On completion of the project and learning experience it was intended that the students would be able to recognise and articulate the impact and sustainability of their design propositions, consider the life-cycle of their proposals and the values these might create for the intended users, communities and contexts. The project was completed in January 2020, as the Covid-19 pandemic was just beginning its spread around the world. This unprecedented catastrophe reinforced the importance of supporting those most in need – the citizens of developing regions in the Global South. In April 2020, the heads of all the UN’s major agencies issued an open letter warning of the risks the virus posed to the world’s most vulnerable countries. It called on wealthier nations to increase funding and help to tackle issues such as the cessation of aid as a result of cancelled flights and disrupted supply routes. These and many other concerns highlighted during the crisis are among the topics explored in this project, which feels even more relevant and urgent than when it was initiated in the summer of 2019. One of the most significant societal shifts currently taking place within the field of sustainable development work is its transformation from being understood as a process of growth or, at its most benign, poverty alleviation, to one of community empowerment and civic participation. The public’s role is developing beyond once-passive community members and recipients of aid, into stakeholders valued for their local knowledge, lived experiences, participation in development projects, and contribution towards policy-making and decision-making. This new dynamic is changing the traditional North-South relationship and holds the potential to challenge the geopolitical hegemony of International Development. The impetus for this shift is a decolonial, collaborative approach to development, research and practice; increased local empowerment, and sustainable solutions to problems that are co-created in context with those affected by and affecting the issue in question. This project asked students to consider what happens in this global landscape ten years from now where Sustainable Development has evolved to the extent that new forms of work and communities of practice transform how people engage, learn and interact with each other, with stakeholders and with the global community around them. The brief gave students the opportunity to explore the underlying complexities regarding sustainable futures, the post-colonial dynamic between ‘norths’ and ‘souths’, post-capitalism and human agency, to envision a future world context, develop it as an experiential exhibit, and produce the designed products, services and experiences for the people who might live and work within it. The project was divided into two sections: The first was a collaborative stage where groups of students were assigned a specific domain to collectively research one aspect of the project challenge, these domains included; Health, Energy, Mobility, Economies, Education, Societal Structures and Environment. Each of these domains were examined through the lenses of Social, Technological, Economic, Ethical, Educational, Values, Political, Legal and Ecological (STEEEVPLE) and were tailored in use, as appropriate per domain. The groups focused on researching and exploring their specific domain and gathering as much information and understanding while working with the external experts to further their knowledge. This group stage culminated in a series of Future World exhibits which tangibly manifest the cohort’s collective knowledge and collaborative understanding of what the future could look like in 10 years from now, after exploring the possible consequences of current actions. The second stage saw students explore their individual response to the Future World that had been created in the first stage. Each student developed their own response to the research by iteratively creating a design outcome that was appropriate to the subject matter. This culminated in each student producing a designed product, service or system and a visual communication of the future experience which they had created. A visual summary of the journey and stages (Project Journey Map) is included within the repository and outlines the collaborative process of designing and the innovative nature of the project’s pedagogical model. The project aims to reveal and address the emerging possibilities collaboratively created by Sustainable Development professionals and designers interacting and learning from each other, to present preferable futures which reveal socio-ecological innovations in development work with the Global South in the near future. The deposited materials are arranged as follows: Readme files - two readme files relate to stage one and stage two of the project as outlined above. Project Journey Map - gives a visual overview of the pedagogical structure and timeline of the project. Data folders - the data folders for stage one of the project are named by the domains through which each group explored possible futures. The data folders for stage two of the project are named for the individual students who conducted the work

Impact of opioid-free analgesia on pain severity and patient satisfaction after discharge from surgery: multispecialty, prospective cohort study in 25 countries

Background: Balancing opioid stewardship and the need for adequate analgesia following discharge after surgery is challenging. This study aimed to compare the outcomes for patients discharged with opioid versus opioid-free analgesia after common surgical procedures.Methods: This international, multicentre, prospective cohort study collected data from patients undergoing common acute and elective general surgical, urological, gynaecological, and orthopaedic procedures. The primary outcomes were patient-reported time in severe pain measured on a numerical analogue scale from 0 to 100% and patient-reported satisfaction with pain relief during the first week following discharge. Data were collected by in-hospital chart review and patient telephone interview 1 week after discharge.Results: The study recruited 4273 patients from 144 centres in 25 countries; 1311 patients (30.7%) were prescribed opioid analgesia at discharge. Patients reported being in severe pain for 10 (i.q.r. 1-30)% of the first week after discharge and rated satisfaction with analgesia as 90 (i.q.r. 80-100) of 100. After adjustment for confounders, opioid analgesia on discharge was independently associated with increased pain severity (risk ratio 1.52, 95% c.i. 1.31 to 1.76; P &lt; 0.001) and re-presentation to healthcare providers owing to side-effects of medication (OR 2.38, 95% c.i. 1.36 to 4.17; P = 0.004), but not with satisfaction with analgesia (beta coefficient 0.92, 95% c.i. -1.52 to 3.36; P = 0.468) compared with opioid-free analgesia. Although opioid prescribing varied greatly between high-income and low- and middle-income countries, patient-reported outcomes did not.Conclusion: Opioid analgesia prescription on surgical discharge is associated with a higher risk of re-presentation owing to side-effects of medication and increased patient-reported pain, but not with changes in patient-reported satisfaction. Opioid-free discharge analgesia should be adopted routinely

Masculinity, morality and hunting, c.1850-1950

The moral imperatives of hunting have long been associated with masculinity. In Britain's age of High Imperialism, hunting assumed greater importance as a 'right of passage' necessary for personal assertion and imperial stability. For proponents, killing wildlife for sport was a 'natural' process which illustrated the ethnocentric and cultural superiority of British upper class men over various "others". This 'pre-eminence' manifested itself through 'male' institutions including elite education, church, army and hunting clubs and found wider expression through hunting books and museums. Dissidents from this pleasing masculinity railed at the celebration of hunting as 'character' training, in particular, the 'objectification' of animals for 'sport'. The ubiquity of the hunting image in the age of High Imperialism, however, was evidence of hunting's popularity as a 'maker of men' despite the concerns of humanitarians.The moral imperatives of hunting have long been associated with masculinity. In Britain's age of High Imperialism, hunting assumed greater importance as a 'right of passage' necessary for personal assertion and imperial stability. For proponents, killing wildlife for sport was a 'natural' process which illustrated the ethnocentric and cultural superiority of British upper class men over various "others". This 'pre-eminence' manifested itself through 'male' institutions including elite education, church, army and hunting clubs and found wider expression through hunting books and museums. Dissidents from this pleasing masculinity railed at the celebration of hunting as 'character' training, in particular, the 'objectification' of animals for 'sport'. The ubiquity of the hunting image in the age of High Imperialism, however, was evidence of hunting's popularity as a 'maker of men' despite the concerns of humanitarians