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Abstract Background Professionalism is expected of health professionals and advocated by professional regulators in the United Kingdom (UK). Concepts of professionalism have evolved in sociological discourse and its meaning for dentistry is unclear. It is, none-the-less, considered a core domain of dental education and professional practice by the United Kingdom regulator, the General Dental Council. This paper reports the sense-making process, or social process, of professionalism in practice within England. Aim To explore the research question ‘What does dental professionalism mean in practice? Methods Taking a constructivist grounded theory approach, involving purposive and theoretical sampling, 24 dental professionals were recruited to participate in this qualitative study. In-depth, semi-structured interviews were conducted by one interviewer (AT). Interviews were recorded, transcribed verbatim, and analysed leading to the development of a theory grounded in the data. Results A focus on the social-professional constructs used by participants to make sense of their experiences, resulted in a grounded theory where Reconciling Duty emerged as the core category. This represents a process of meeting professional duties to different parties that are often mutually exclusive. It is comprised of three supporting categories: Applying order to the system, where individuals attempt to identify what constitutes professional attitudes and behaviours, Rationalising what is fair, where individuals make judgements on how the conflict between duties should be resolved, and finally Responding to the System, where individuals attempt to actualise these desired resolutions in the context of the complex social system in which they practice. Three dentist archetypes (typologies) emerged, which involved a personal (Type 1), patient (Type 2), or a societal (Type 3) compromise. Conclusion Professionalism can be conceptualised as process of reconciling multiple, competing, legitimate duties to different parties, in seeking a fair solution. Once this has been identified, individuals need to work within the complex system of dentistry to make their identified outcome a reality. The findings suggest that using the theory of Reconciling Duty helps us to engage with the meaning that the participants drew from the term ‘professionalism’, and anchors it in the lived, everyday professional experiences and challenges faced. A novel typology is proposed, commensurate with calls for a systems approach to the topic.

BackgroundGlobally, there is growing attention towards health system strengthening, and the importance of quality in health systems. However, fragile and conflict-affected states present particular challenges. The aim of this study was to explore health system strengthening in fragile and conflict-affected states by synthesising the evidence from published literature.MethodsWe conducted a review of systematic reviews (Prospero Registration Number: CRD42022371955) by searching Ovid (Medline, Embase, and Global Health), Scopus, Web of Science, and the Cochrane Library databases. Only English-language publications were considered. The Joanna Briggs Institute (JBI) Critical Appraisal Tool was employed to assess methodological quality of the included studies. The findings were narratively synthesised and presented in line with the Lancet's 'high-quality health system framework'.ResultsTwenty-seven systematic reviews, out of 2,704 identified records, considered key dimensions of health systems in fragile and conflict-affected states, with the 'foundations' domain having most evidence. Significant challenges to health system strengthening, including the flight of human capital due to safety concerns and difficult working conditions, as well as limited training capacities and resources, were identified. Facilitators included community involvement, support systems and innovative financing mechanisms. The importance of coordinated and integrated responses tailored to the context and stage of the crisis situation was emphasised in order to strengthen fragile health systems. Overall, health system strengthening initiatives included policies encouraging the return and integration of displaced healthcare workers, building local healthcare workers capacity, strengthening education and training, integrating healthcare services, trust-building, supportive supervision, and e-Health utilisation.ConclusionThe emerging body of evidence on health system strengthening in fragile and conflict-affected states highlights its complexity. The findings underscore the significance of adopting a comprehensive approach and engaging various stakeholders in a coordinated manner considering the stage and context of the situation.

Introduction: The Global Strategy for Human Resources for Health 2030, requires member states to half their dependency on an international workforce by 2030. In order to design policies towards that goal, country-specific research on migration motivations of the health workforce is required. The United Kingdom (UK) is a net importer of health professionals and whilst there is a body of research on doctors’ and nurses’ migration, there is no research on the migration motivations of migrant dentists in the UK. This research explored the migration motivations of internationally qualified dentists (IQDs) in the UK and presents a typology to understand the global migration of dentists in the context of oral health workforce. Methods: The paper presents qualitative data from semi-structured interviews conducted between August 2014 and October 2017, of IQDs working in the United Kingdom. The topic guide for interviews was informed by the literature, with new themes added inductively. A phenomenological approach involving an epistemological stance of interpretivism, was used with framework analysis. Results: A total of 38 internationally qualified dentists (M = 18, F = 20), migrating from the five World Health Organization regions, and working in general practice, NHS hospitals and in community dental services across the four nations of the UK were interviewed. Seven types of internationally qualified dentists were identified working in the UK. They were livelihood migrants, career-orientated migrants, dependant migrants, backpacker migrants, commuter migrants, undocumented migrants, and education-tourist migrant. The categories were based on their migration motivations, which were complex, multifactorial, and included personal, professional, national, and international drivers. The typology, based on their migration motivations, offered a structured, comprehensive understanding of the migrant dental workforce. This typology involving dentists provides additional dimensions to commuter and undocumented migrants described in the context of other health professionals. The education-tourist migrant is a new category proposed as an extension to existing typology in health professional migration. Conclusions: The typology of internationally qualified dentists has congruency with other health professionals’ typology in categories previously described and demonstrates that each of these categories are complex, fluid and change in response to policy changes. The new category of education-tourist migrant along with oral health dimensions of commuter and undocumented migrants adds to the existing typology in health professional migration.

Few descriptions of using the femoral vein as an arterial replacement exist in the pediatric surgical literature. A 3-year-old child sustained a gunshot wound to the right subclavian artery, which resulted in ischemia to the upper extremity. The left femoral vein was harvested and used to reconstruct the subclavian artery. This report highlights a rare case of subclavian artery reconstruction using a femoral vein conduit in a young child after a penetrating injury.

BackgroundComparative optimism, the belief that negative events are more likely to happen to others rather than to oneself, is well established in health risk research. It is unknown, however, whether comparative optimism also permeates people's health expectations and potentially behaviour during the COVID-19 pandemic.ObjectivesData were collected through an international survey (N = 6485) exploring people's thoughts and psychosocial behaviours relating to COVID-19. This paper reports UK data on comparative optimism. In particular, we examine the belief that negative events surrounding risk and recovery from COVID-19 are perceived as more likely to happen to others rather than to oneself.MethodsUsing online snowball sampling through social media, anonymous UK survey data were collected from N = 645 adults during weeks 5-8 of the UK COVID-19 lockdown. The sample was normally distributed in terms of age and reflected the UK ethnic and disability profile.FindingsRespondents demonstrated comparative optimism where they believed that as compared to others of the same age and gender, they were unlikely to experience a range of controllable (eg accidentally infect/ be infected) and uncontrollable (eg need hospitalization/ intensive care treatment if infected) COVID-19-related risks in the short term (P

The sustained governmental assault on the National Health Service (NHS) in England during post-1970s financialised or rentier capitalism has received considerable attention by the research community. There is some evidence, however, that many of those members of the public who have not had occasion to use the NHS remain largely ill-informed about the extent of, and reasons for, its present troubles. In this paper I offer an auto/biographic account of my own recent experiences as a patient with type 2 diabetes and subsequent polymyalgia in both primary and secondary care. I then deploy analytic induction to consider, and explain, my personal travails against the background of the shifting nature of doctor-patient interaction occasioned by governmental politics in relation to the NHS. The result is an illustrated story of the decline of health care at a political juncture when the ever-expanding capital assets of a tiny minority of the population trumps the health care needs of the population as a whole. The present impoverishment of management and care must be understood with reference to wider aspects of macro-social change. The paper concludes with some ideas about how to (re)fund a severely ailing NHS.

Abstract Background There is a social expectation that dentists demonstrate professionalism. Although the General Dental Council puts it at the heart of their regulatory agenda, there is not yet consensus on the meaning and implications of the term. Objective To explore practising dentists’ understanding of the character traits commonly associated with professionalism and what these mean in practice. Method Constructivist grounded theory was employed throughout this study. Qualitative, in-depth interviews were conducted with dental professionals in England recruited through theoretical sampling to saturation point. Interviews used a topic guide informed by the literature, and analysis was conducted through constant comparison during data collection. Results The study found that traits commonly associated with professionalism in the literature were difficult for dentists to define clearly or operationalise in a clinical setting. There was disagreement over how some traits should be understood, and it was unclear to participants how, or indeed if, the listed traits were directly relevant to practice in their current form. Conclusion Rather than expecting unconditional adherence to an externally imposed definition, further exploration is required to understand how health professionals make sense of professionalism by reference to their lived experiences and worldviews. In Brief Institutional expectations of professionalism, defined through character traits and behaviours, do not appear to map neatly on to the experiences of dental professionals. Straightforward, apparently uncontroversial terms elicited a wide range of responses, including disagreement. This brought in to question whether achieving consensus is possible. Analysing how our respondents understood the terms by reference to the meanings they constructed from lived experience offers deeper insights.

Abstract Introduction Regulatory processes for Oral health care professionals are considered essential for patient safety and to ensure health workforce quality. The global variation in their registration and regulation is under-reported in the literature. Regulatory systems could become a barrier to their national and international movement, leading to loss of skilled human resources. The General Dental Council is the regulatory authority in the UK, one of the nine regulators of health care overseen by the Professional Standards Authority. Aim The aim of this paper is to present the professional integration experiences of internationally qualified dentists (IQDs) working in the UK, against the background of regulation and accreditation nationally. Methods Registration data were obtained from the General Dental Council to inform the sampling and recruitment of research participants. Semi-structured interviews of 38 internationally qualified dentists working in the United Kingdom were conducted between August 2014 and October 2017. The topic guide which explored professional integration experiences of the dentists was informed by the literature, with new themes added inductively. A phenomenological approach involving an epistemological stance of interpretivism, was used with framework analysis to detect themes. Results Internationally qualified dentist’s professional integration was influenced by factors that could be broadly classified as structural (source country training; registration and employment; variation in practising dentistry) and relational (experiences of discrimination; value of networks and support; and personal attributes). The routes to register for work as a dentist were perceived to favour UK dental graduates and those qualifying from the European Economic Area. Dentists from the rest of the world reported experiencing major hurdles including succeeding in the licensing examinations, English tests, proving immigration status and succeeding in obtaining a National Health Service performer number, all prior to being able to practice within state funded dental care. Conclusion The pathways for dentists to register and work in state funded dental care in UK differ by geographic type of registrant, creating significant inconsistencies in their professional integration. Professional integration is perceived by an individual IQD as a continuum dictated by host countries health care systems, workforce recruitment policies, access to training, together with their professional and personal skills. The reliance of the UK on internationally qualified dentists has increased in the past two decades, however, it is not known how these trends will be affected by UK’s exit from the European Union and the COVID-19 pandemic.

The chemokine SDF-1 (CXCL12) and its receptor CXCR4 control several processes during embryonic development such as the regulation of stem cell proliferation, differentiation, and migration. However, the role of this pathway in the formation of the pituitary gland is not understood. We sought to characterise the expression patterns of CXCR4, SDF-1 and CXCR7 at different stages of pituitary gland development. Our expression profiling revealed that SDF-1 is expressed in progenitor-rich regions of the pituitary anterior lobe, that CXCR4 and CXCR7 have opposite expression domains and that CXCR4 expression is conserved between mice and human embryos. We then assessed the importance of this signalling pathway in the development and function of the murine pituitary gland through conditional deletion of CXCR4 in embryonic pituitary progenitors. Successful and specific ablation of CXCR4 expression in embryonic pituitary progenitors did not lead to observable embryonic nor postnatal defects but allowed the identification of stromal CXCR4+ cells not derived from HESX1+ progenitors. Further analysis of constitutive SDF-1, CXCR7 and CXCR4 mutants of the pathway indicates that CXCR4 expression in HESX1+ cells and their descendants is not essential for normal pituitary development in mice.

Abstract Aim To review evidence on oral health practices, beliefs/views and experiences of community-dwelling older adults living with dementia, including their carers. Materials and methods A search of key terms across six databases including Pubmed, Web of Science and OVID (Embase, MEDLINE [R] and PsycINFO) and Google Scholar was conducted, supplemented by reference screening. The Mixed Methods Appraisal Tool (MMAT) 2018 was used to assess the methodological quality. Results Eighteen studies reported across 19 papers were included in the review. Papers largely focused on normative needs (n = 13), whilst also reporting oral health-related experiences (n = 2), practices (n = 7), and beliefs/views (n = 9), of community dwellers with dementia. Generally, people living with dementia presented with poor oral and dental health, the exception being one study where dental care was integrated with memory clinic services. Maintenance of oral health focused only on toothbrushing. Overall, people living with dementia have reduced capacity for self-performed oral hygiene and high reliance on caregivers. There was a paucity of evidence on their perceptions of oral health and quality of life, the findings of which were equivocal, with weak evidence suggesting possible difficulty in identifying and communicating their needs. Experiences of accessing dental care, when explored, appear to be system dependent. Conclusion There was limited research evidence on oral health-related practices, beliefs/views and experiences of people with dementia. Recommendations for future research are presented.

In this wide-ranging interview Graham Scambler provides an overview of his long academic career. He discusses how he became a medical sociologist, his early work on epilepsy and stigma, his part in the development of sociology textbooks for medical students, the diversity of his work and his many collaborations, his 'theoretical turn', his longstanding interest in critical realism and his attitude to 'bridge building' between philosophy and empirical work. [ABSTRACT FROM AUTHOR]

We examined perceived self-other differences (self-uniqueness) in appraisals of one’s risk of an infectious disease (COVID-19), one’s adherence to behavioural precautionary measures against the disease, and the impact of these measures on one’s life. We also examined the relationship of self-uniqueness with information seeking and trust in sources of information about the disease. We administered an online survey to a community sample (N = 8696) of Dutch-speaking individuals, mainly in Belgium and The Netherlands, during the first lockdown (late April-Mid June 2020). As a group, participants reported that they were less likely to get infected or infect others or to suffer severe outcomes than average (unrealistic optimism) and that they adhered better than average to behavioural precautionary measures (illusory superiority). Except for participants below 25, who reported that they were affected more than average by these measures (egocentric impact bias), participants also generally reported that they were less affected than average (allocentric impact bias). Individual differences in self-uniqueness were associated with differences in the number of information sources being used and trust on these sources. Higher comparative optimism for infection, self-superiority, and allocentric impact perception were associated with information being sought from fewer sources; higher self-superiority and egocentric impact perception were associated with lower trust. We discuss implications for health communication.

In this paper I ground a brief account of the impact of COVID-19 on the United Kingdom in an understanding of a decade of austerity politics from 2010 to 2020, itself a product of the advent and consolidation of post-1970s financialised or rentier capitalism. I argue that such an analysis is essential if realistic plans are to be laid for a “better”—understood here as a more equitable or “fairer”—society. I go on to consider the contributions that sociology can, and arguably should, make to this end. This involves a range of engagements from scholarship at one end of the spectrum to action or muckraking sociology at the other. In addition to plotting a role for sociology, I suggest a set of criteria for recognizing a “fairer society”; postulate a series of institutional reforms that might characterize the attainment of such a society; and outline and confront social structural, cultural and agential obstacles to its realization. A theme running throughout the paper is that the delineation and promulgation of the “good society” remains central to any credible—that is, post-Enlightenment reconstruction of - the sociological project.

Summary: Cxcl12-null embryos have dysplastic, misaligned, and hyperplastic semilunar valves (SLVs). In this study, we show that CXCL12 signaling via its receptor CXCR4 fulfills distinct roles at different stages of SLV development, acting initially as a guidance cue to pattern cellular distribution within the valve primordia during the endocardial-to-mesenchymal transition (endoMT) phase and later regulating mesenchymal cell proliferation during SLV remodeling. Transient, anteriorly localized puncta of internalized CXCR4 are observed in cells undergoing endoMT. In vitro, CXCR4+ cell orientation in response to CXCL12 requires phosphatidylinositol 3-kinase (PI3K) signaling and is inhibited by suppression of endocytosis. This dynamic intracellular localization of CXCR4 during SLV development is related to CXCL12 availability, potentially enabling activation of divergent downstream signaling pathways at key developmental stages. Importantly, Cxcr7-/- mutants display evidence of excessive CXCL12 signaling, indicating a likely role for atypical chemokine receptor CXCR7 in regulating ligand bioavailability and thus CXCR4 signaling output during SLV morphogenesis.

Primary ciliary dyskinesia (PCD) is a rare, genetically heterogeneous ciliopathy disorder affecting cilia and sperm motility. A range of ultrastructural defects of the axoneme underlie the disease, which is characterised by chronic respiratory symptoms and

Mouse embryo imaging is conventionally carried out on ex vivo embryos excised from the amniotic sac, omitting vital structures and abnormalities external to the body. Here, we present an in amnio MR imaging methodology in which the mouse embryo is retained

Abstract In this study, eight species of Chroogomphus are recognized from Europe: C. britannicus, C. aff. filiformis 1, C. fulmineus, C. cf. helveticus, C. mediterraneus, C. cf. purpurascens, C. rutilus, and C. subfulmineus. Different candidates for the application of the name C. rutilus are evaluated and the best fit to the description is selected; lecto- and epitypes are chosen to fix the name. Chroogomphus fulmineus and C. mediterraneus are also epitypified and a new species, C. subfulmineus, is described. The infrageneric classification is revised and a new subgenus Siccigomphus and three new sections, Confusi, Filiformes, and Fulminei are introduced. The former sections Chroogomphus and Floccigomphus are elevated to subgeneric level. Comparison of the ITS regions (nuc rDNA ITS1-5.8S-ITS2) of all species studied shows that there is a minimum interspecific difference of 1.5 %, with the exception of the two species belonging to sect. Fulminei which differ by a minimum of 0.9 %. Ecological specimen data indicate that species of Chroogomphus form basidiomes under members of Pinaceae, with a general preference for species of Pinus. Five European species have been recorded under Picea, while Abies and Larix have also been recorded as tree associates, although the detailed nutritional relationships of the genus, involving other suilloid fungi in particular, have yet to be fully clarified.

Bidirectional (anterograde and retrograde) motor-based intraflagellar transport (IFT) governs cargo transport and delivery processes that are essential for primary cilia growth and maintenance and for hedgehog signaling functions. The IFT dynein-2 motor complex that regulates ciliary retrograde protein transport contains a heavy chain dynein ATPase/motor subunit, DYNC2H1, along with other less well functionally defined subunits. Deficiency of IFT proteins, including DYNC2H1, underlies a spectrum of skeletal ciliopathies. Here, by using exome sequencing and a targeted next-generation sequencing panel, we identified a total of 11 mutations in WDR34 in 9 families with the clinical diagnosis of Jeune syndrome (asphyxiating thoracic dystrophy). WDR34 encodes a WD40 repeat-containing protein orthologous to Chlamydomonas FAP133, a dynein intermediate chain associated with the retrograde intraflagellar transport motor. Three-dimensional protein modeling suggests that the identified mutations all affect residues critical for WDR34 protein-protein interactions. We find that WDR34 concentrates around the centrioles and basal bodies in mammalian cells, also showing axonemal staining. WDR34 coimmunoprecipitates with the dynein-1 light chain DYNLL1 in vitro, and mining of proteomics data suggests that WDR34 could represent a previously unrecognized link between the cytoplasmic dynein-1 and IFT dynein-2 motors. Together, these data show that WDR34 is critical for ciliary functions essential to normal development and survival, most probably as a previously unrecognized component of the mammalian dynein-IFT machinery.

Previously, we showed that serum and monocytes from patients with CF exhibit an enhanced NLRP3-inflammasome signature with increased IL-18, IL-1β, caspase-1 activity and ASC speck release (Scambler et al. eLife 2019). Here we show that CFTR modulators down regulate this exaggerated proinflammatory response following LPS/ATP stimulation. In vitro application of ivacaftor/lumacaftor or ivacaftor/tezacaftor to CF monocytes showed a significant reduction in IL-18, whereas IL-1β was only reduced with ivacaftor/tezacaftor. Thirteen adults starting ivacaftor/lumacaftor and eight starting ivacaftor/tezacaftor were assessed over three months. Serum IL-18 and TNF decreased significantly with treatments, but IL-1β only declined following ivacaftor/tezacaftor. In (LPS/ATP-stimulated) PBMCs, IL-18/TNF/caspase-1 were all significantly decreased and IL-10 was increased with both combinations. Ivacaftor/tezacaftor alone showed a significant reduction in IL-1β and pro-IL-1β mRNA. This study demonstrates that these CFTR modulator combinations have potent anti-inflammatory properties, in addition to their ability to stimulate CFTR function, which could contribute to improved clinical outcomes.

Heterogeneity of lymphatic vessels during embryogenesis is critical for organ-specific lymphatic function. Little is known about lymphatics in the developing kidney, despite their established roles in pathology of the mature organ. We performed three-dimensional imaging to characterize lymphatic vessel formation in the mammalian embryonic kidney at single-cell resolution. In mouse, we visually and quantitatively assessed the development of kidney lymphatic vessels, remodeling from a ring-like anastomosis under the nascent renal pelvis; a site of VEGF-C expression, to form a patent vascular plexus. We identified a heterogenous population of lymphatic endothelial cell clusters in mouse and human embryonic kidneys. Exogenous VEGF-C expanded the lymphatic population in explanted mouse embryonic kidneys. Finally, we characterized complex kidney lymphatic abnormalities in a genetic mouse model of polycystic kidney disease. Our study provides novel insights into the development of kidney lymphatic vasculature; a system which likely has fundamental roles in renal development, physiology and disease.

Nawaf Al Shammary,1 Koula Asimakopoulou,1 Fraser McDonald,2 Jonathon T Newton,1 Sasha Scambler1 1Division of Population and Patient Health, 2Craniofacial Development and Stem Cell Biology Department, King’s College London Dental Institute, London, UKBackground: Patient adherence in orthodontic treatment is extremely important as it is linked with better treatment outcomes. Despite its importance, however, there is no shared definition of the concept. This makes the recording of adherence-related behaviors in patient notes difficult. The current study explored how, and to what extent adherence is recorded in adult patients’ medical records by orthodontists working in a large National Health Service (NHS) London hospital.Materials and methods: A mixed-methods approach was used. A total of 17 clinicians with a mean age of 31 years (SD =4.87) provided N=20 case notes spanning N=324 appointments with patients they judged to be non-adherent. The notes were inspected for evidence of recording of patient adherence using adherence indicators identified in the literature.Results: The term “adherence” did not feature in any notes. The quantitative analysis showed that the three most frequent adherence-related behaviors recorded in notes were “oral hygiene,” “appointment attendance” and “breakages of appliances.” Qualitative analysis not only confirmed these factors but also showed that 1) the clinical aspects of treatment, 2) clinician–patient interaction factors and 3) patient attitudes also featured. This part of the analysis also highlighted inconsistencies across case notes in terms of the amount of information being recorded.Conclusion: Adherence as a term does not feature in the clinical case notes of clinician-identified non-adherent adult patients, while predictors of adherence are recorded with varying degrees of consistency. Keywords: adherence, adults, orthodontic treatment

Abstract In many different cell types, pro-inflammatory agonists induce the expression of cyclooxygenase 2 (COX-2), an enzyme that catalyzes rate-limiting steps in the conversion of arachidonic acid to a variety of lipid signaling molecules, including prostaglandin E2 (PGE2). PGE2 has key roles in many early inflammatory events, such as the changes of vascular function that promote or facilitate leukocyte recruitment to sites of inflammation. Depending on context, it also exerts many important anti-inflammatory effects, for example increasing the expression of the anti-inflammatory cytokine interleukin 10 (IL-10), and decreasing that of the pro-inflammatory cytokine tumor necrosis factor (TNF). The tight control of both biosynthesis of, and cellular responses to, PGE2 are critical for the precise orchestration of the initiation and resolution of inflammatory responses. Here we describe evidence of a negative feedback loop, in which PGE2 augments the expression of dual specificity phosphatase 1, impairs the activity of mitogen-activated protein kinase p38, increases the activity of the mRNA-destabilizing factor tristetraprolin, and thereby inhibits the expression of COX-2. The same feedback mechanism contributes to PGE2-mediated suppression of TNF release. Engagement of the DUSP1-TTP regulatory axis by PGE2 is likely to contribute to the switch between initiation and resolution phases of inflammation.

Cystic Fibrosis (CF) is a monogenic disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, resulting in defective CFTR-mediated chloride and bicarbonate transport, with dysregulation of epithelial sodium channels (ENaC). These changes alter fluid and electrolyte homeostasis and result in an exaggerated proinflammatory response driven, in part, by infection. We tested the hypothesis that NLRP3 inflammasome activation and ENaC upregulation drives exaggerated innate-immune responses in this multisystem disease. We identify an enhanced proinflammatory signature, as evidenced by increased levels of IL-18, IL-1β, caspase-1 activity and ASC-speck release in monocytes, epithelia and serum with CF-associated mutations; these differences were reversed by pretreatment with NLRP3 inflammasome inhibitors and notably, inhibition of amiloride-sensitive sodium (Na+) channels. Overexpression of β-ENaC, in the absence of CFTR dysfunction, increased NLRP3-mediated inflammation, indicating that dysregulated, ENaC-dependent signalling may drive exaggerated inflammatory responses in CF. These data support a role for sodium in modulating NLRP3 inflammasome activation.

Cystic Fibrosis (CF) is a recessive genetic disorder caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR). CFTR mutations cause dysregulation of channel function with intracellular accumulation of misfolded proteins and endoplasmic reticulum (ER) stress, with activation of the IRE1α-XBP1 pathway that regulates a subset of unfolded protein response (UPR) genes. This pathway regulates a group of genes that control proinflammatory and metabolic responses in different immune cells; however, the metabolic state of immune cells and the role of this pathway in CF remain elusive. Our results indicate that only innate immune cells from CF patients present increased levels of ER stress, mainly affecting neutrophils, monocytes, and macrophages. An overactive IRE1α-XBP1 pathway reprograms CF M1 macrophages toward an increased metabolic state, with increased glycolytic rates and mitochondrial function, associated with exaggerated production of TNF and IL-6. This hyper-metabolic state, seen in CF macrophages, is reversed by inhibiting the RNase domain of IRE1α, thereby decreasing the increased glycolic rates, mitochondrial function and inflammation. Altogether, our results indicate that innate immune cells from CF patients are primarily affected by ER stress. Moreover, the IRE1α-XBP1 pathway of the UPR is responsible for the hyper-metabolic state seen in CF macrophages, which is associated with the exaggerated inflammatory response. Modulating ER stress, metabolism and inflammation, by targeting IRE1α, may improve the metabolic fitness of macrophages, and other immune cells in CF and other immune-related disorders.

TNFAIP3 encodes the NF-κB regulatory protein A20. High-penetrance heterozygous mutations in TNFAIP3 cause a haploinsufficiency of A20 (HA20), inadequate inhibition of NF-κB pathway, and an early onset autoinflammatory disorder. However, the clinical phenotype of patients with HA20 varies greatly and clinical diagnoses prior to establishing the genetic cause, included both autoimmune and autoinflammatory conditions. Here, we present the first patient with HA20, who was previously diagnosed with AOSD but was later found to have a novel heterozygous variant in TNFAIP3 and who was successfully treated with anti-IL6 receptor biologic tocilizumab (RoActemra). We discovered a novel heterozygous mutation in TNFAIP3 c.1906C>T, not previously found in ExAC database. Further analysis shows that this single-nucleotide variant at the terminal residue of TNFAIP3 exon 7 produces an alternatively spliced mRNA resulting in p.His636fsTer1. Additional genetic analysis of family members shows that this variant does segregate with the inflammatory clinical phenotypes. Subsequent functional test show that NF-κB activation, measured as intracellular phosphorylation of p65 in CD14 + monocytes, was more enhanced in the patient compared with healthy controls (HC) following stimulation with LPS. This was associated with higher production of inflammatory cytokines by the patients PBMC in response to LPS and ATP and enhanced activation of NLRP3 inflammasome complex. Furthermore, increased activation of NLRP3 inflammasome was evident systemically, since we detected higher levels of ASC specks in patients’ sera compared with HC. Finally, we used population genetics data from GnomAD to construct a map of both genetic conservation and most probable disease-causing variants in TNFAIP3 which might be found in future cases of HA20.

Tumor necrosis factor (TNF)-receptor-associated periodic fever syndrome (TRAPS) is a rare monogenic autoinflammatory disorder characterized by mutations in the TNFRSF1A gene, causing TNF-receptor 1 (TNFR1) misfolding, increased cellular stress, activation of the unfolded protein response (UPR), and hyperresponsiveness to lipopolysaccharide (LPS). Both microRNA (miR)-146a and miR-155 provide negative feedback for LPS-toll-like receptor 2/4 signaling and cytokine production, through regulation of nuclear factor kappa B (NF-κB). In this study, we hypothesized that proinflammatory cytokine signaling in TRAPS downregulates these two miRs, resulting in LPS-induced hyperresponsiveness in TRAPS dermal fibroblasts (DFs), irrespective of the underlying genetic mutation. Primary DF were isolated from skin biopsies of TRAPS patients and healthy controls (HC). TNFR1 cell surface expression was measured using immunofluorescence. DF were stimulated with LPS, interleukin (IL)-1β, thapsigargin, or TNF, with and without inositol-requiring enzyme 1 (IRE1) inhibitor (4u8C), following which miR-146a and miR-155 expression was measured by RT-qPCR. IL-1β, IL-6, and TNF secretion was measured by enzyme-linked immunosorbent assays, and baseline expression of 384 different miRs was assessed using microfluidics assays. TNFR1 was found to be expressed on the surface of HC DF but expression was deficient in all samples with TRAPS-associated mutations. HC DF showed significant dose-dependent increases in both miR-146a and miR-155 expression levels in response to LPS; however, TRAPS DF failed to upregulate either miR-146a or miR-155 under the same conditions. This lack of miR-146a and miR-155 upregulation was associated with increased proinflammatory cytokine production in TRAPS DF in response to LPS challenge, which was abrogated by 4u8C. Incubation of HC DF with IL-1β led to downregulation of miR-146a and miR-155 expression, which was dependent on IRE1 enzyme. We observed global dysregulation of hundreds of other miRs at baseline in the TRAPS DF. In summary, these data suggest a mechanism whereby IL-1β, produced in response to activation of the UPR in TRAPS DF, downregulates miR-146a and miR-155, by inducing IRE1-dependent cleavage of both these miRs, thereby impairing negative regulation of NF-κB and increasing proinflammatory cytokine production.

The CXCL12-CXCR4 pathway has crucial roles in stem cell homing and maintenance, neuronal guidance, cancer progression, inflammation, remote-conditioning, cell migration and development. Recently, work in chick suggested that signalling via CXCR4 in neural crest cells (NCCs) has a role in the 22q11.2 deletion syndrome (22q11.2DS), a disorder where haploinsufficiency of the transcription factor TBX1 is responsible for the major structural defects. We tested this idea in mouse models. Our analysis of genes with altered expression in Tbx1 mutant mouse models showed down-regulation of Cxcl12 in pharyngeal surface ectoderm and rostral mesoderm, both tissues with the potential to signal to migrating NCCs. Conditional mutagenesis of Tbx1 in the pharyngeal surface ectoderm is associated with hypo/aplasia of the 4th pharyngeal arch artery (PAA) and interruption of the aortic arch type B (IAA-B), the cardiovascular defect most typical of 22q11.2DS. We therefore analysed constitutive mouse mutants of the ligand (CXCL12) and receptor (CXCR4) components of the pathway, in addition to ectodermal conditionals of Cxcl12 and NCC conditionals of Cxcr4. However, none of these typical 22q11.2DS features were detected in constitutively or conditionally mutant embryos. Instead, duplicated carotid arteries were observed, a phenotype recapitulated in Tie-2Cre (endothelial) conditional knock outs of Cxcr4. Previous studies have demonstrated genetic interaction between signalling pathways and Tbx1 haploinsufficiency e.g. FGF, WNT, SMAD-dependent. We therefore tested for possible epistasis between Tbx1 and the CXCL12 signalling axis by examining Tbx1 and Cxcl12 double heterozygotes as well as Tbx1/Cxcl12/Cxcr4 triple heterozygotes, but failed to identify any exacerbation of the Tbx1 haploinsufficient arch artery phenotype. We conclude that CXCL12 signalling via NCC/CXCR4 has no major role in the genesis of the Tbx1 loss of function phenotype. Instead, the pathway has a distinct effect on remodelling of head vessels and interventricular septation mediated via CXCL12 signalling from the pharyngeal surface ectoderm and second heart field to endothelial cells.