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Purpose The purpose of this study was to elucidate the role and molecular consequences of impaired glutathione (GSH) biosynthesis on eye development. Methods GSH biosynthesis was impaired in surface ectoderm-derived ocular tissues by crossing Gclcf/f mice with hemizygous Le-Cre transgenic mice to produce Gclcf/f/Le-CreTg/- (KO) mice. Control mice included Gclcf/f and Gclcwt/wt/Le-CreTg/- mice (CRE). Eyes from all mice (at various stages of eye development) were subjected to histological, immunohistochemical, Western blot, RT-qPCR, RNA-seq, and subsequent Gene Ontology, Ingenuity Pathway Analysis and TRANSFAC analyses. PAX6 transactivation activity was studied using a luciferase reporter assay in HEK293T cells depleted of GSH using buthionine sulfoximine (BSO). Results Deletion of Gclc diminished GSH levels, increased reactive oxygen species (ROS), and caused an overt microphthalmia phenotype characterized by malformation of the cornea, iris, lens, and retina that is distinct from and much more profound than the one observed in CRE mice. In addition, only the lenses of KO mice displayed reduced crystallin (α, β), PITX3 and Foxe3 expression. RNA-seq analyses at postnatal day 1 revealed 1552 differentially expressed genes (DEGs) in the lenses of KO mice relative to those from Gclcf/f mice, with Crystallin and lens fiber cell identity genes being downregulated while lens epithelial cell identity and immune response genes were upregulated. Bioinformatic analysis of the DEGs implicated PAX6 as a key upstream regulator. PAX6 transactivation activity was impaired in BSO-treated HEK293T cells. Conclusions These data suggest that impaired ocular GSH biosynthesis may disrupt eye development and PAX6 function.

This study piloted the use of ACT-informed exposure as an adjunct to habit reversal training (HRT) for excoriation disorder (ExD). Using a nonconcurrent multiple baseline single case design, four participants completed sessions of exposure and HRT. Repeated measures and self-report data were collected on skin picking and psychological flexibility. Two participants completed HRT followed by exposure, and two participants completed exposure followed by HRT. Results support the effectiveness of HRT in reducing picking. Results suggest exposure may have some impact in reducing picking, but effects were weaker compared to HRT. Contrary to predictions, repeated measures and self-report data did not indicate consistent improvement in psychological flexibility during exposure phases. As any reduction in picking may be clinically meaningful and all participants maintained gains at follow-up, there is some indication that exposure may be a second-line treatment worth further study. Limitations and future areas of research are discussed.

MotivationHigh-dimension cytometry assays can simultaneously measure dozens of markers, enabling the investigation of complex phenotypes. However, as manual gating relies on previous biological knowledge, few marker combinations are often assessed. This results in complex phenotypes with potential for biological relevance being overlooked. Here we present PhenoComb, an R package that allows agnostic exploration of phenotypes by assessing all combinations of markers.DesignPhenoComb uses signal intensity thresholds to assign markers to discrete states (e.g. negative, low, high) and then counts the number of cells per sample from all possible marker combinations in a memory-safe manner. Time and disk space are the only constraints on the number of markers evaluated. PhenoComb also provides several approaches to perform statistical comparisons, evaluate the relevance of phenotypes, and assess the independence of identified phenotypes. PhenoComb allows users to guide analysis by adjusting several function arguments such as identifying parent populations of interest, filtering of low-frequency populations, and defining a maximum complexity of phenotypes to evaluate. We have designed PhenoComb to be compatible with local computer or server-based use.ResultsIn testing of PhenoComb’s performance on synthetic datasets, computation on 16 markers was completed in the scale of minutes and up to 26 markers in hours. We applied PhenoComb to two publicly available datasets: an HIV flow cytometry dataset (12 markers and 421 samples) and the COVIDome CyTOF dataset (40 markers and 99 samples). In the HIV dataset, PhenoComb identified immune phenotypes associated with HIV seroconversion, including those highlighted in the original publication. In the COVID dataset, we identified several immune phenotypes with altered frequencies in infected individuals relative to healthy individuals. Collectively, PhenoComb represents a powerful discovery tool for agnostically assessing high-dimension, single-cell data.AvailabilityThe PhenoComb R package can be downloaded from https://github.com/SciOmicsLab/PhenoComb

We investigate how humans perform the task of dubbing video content from one language into another, leveraging a novel corpus of 319.57 hours of video from 54 professionally produced titles. This is the first such large-scale study we are aware of. The results challenge a number of assumptions commonly made in both qualitative literature on human dubbing and machine-learning literature on automatic dubbing, arguing for the importance of vocal naturalness and translation quality over commonly emphasized isometric (character length) and lip-sync constraints, and for a more qualified view of the importance of isochronic (timing) constraints. We also find substantial influence of the source-side audio on human dubs through channels other than the words of the translation, pointing to the need for research on ways to preserve speech characteristics, as well as semantic transfer such as emphasis/emotion, in automatic dubbing systems., Comment: Accepted at TACL. pre-MIT Press publication version

Motivation High-dimensional cytometry assays can simultaneously measure dozens of markers, enabling the investigation of complex phenotypes. However, as manual gating relies on previous biological knowledge, few marker combinations are often assessed. This results in complex phenotypes with the potential for biological relevance being overlooked. Here, we present PhenoComb, an R package that allows agnostic exploration of phenotypes by assessing all combinations of markers. PhenoComb uses signal intensity thresholds to assign markers to discrete states (e.g. negative, low, high) and then counts the number of cells per sample from all possible marker combinations in a memory-safe manner. Time and disk space are the only constraints on the number of markers evaluated. PhenoComb also provides several approaches to perform statistical comparisons, evaluate the relevance of phenotypes and assess the independence of identified phenotypes. PhenoComb allows users to guide analysis by adjusting several function arguments, such as identifying parent populations of interest, filtering of low-frequency populations and defining a maximum complexity of phenotypes to evaluate. We have designed PhenoComb to be compatible with a local computer or server-based use. Results In testing of PhenoComb’s performance on synthetic datasets, computation on 16 markers was completed in the scale of minutes and up to 26 markers in hours. We applied PhenoComb to two publicly available datasets: an HIV flow cytometry dataset (12 markers and 421 samples) and the COVIDome CyTOF dataset (40 markers and 99 samples). In the HIV dataset, PhenoComb identified immune phenotypes associated with HIV seroconversion, including those highlighted in the original publication. In the COVID dataset, we identified several immune phenotypes with altered frequencies in infected individuals relative to healthy individuals. Collectively, PhenoComb represents a powerful discovery tool for agnostically assessing high-dimensional single-cell data. Availability and implementation The PhenoComb R package can be downloaded from https://github.com/SciOmicsLab/PhenoComb. Supplementary information Supplementary data are available at Bioinformatics Advances online.

Clinical trials during public health emergencies of novel medical products such as therapeutics and vaccines in resource-limited settings are daunting due to the limited capacity for regulatory assessment. Regulating clinical trials during the Ebola outbreak in Sierra Leone required expedited evaluation to identify medical products that could be promptly introduced to combat the epidemic in the absence of approved treatment or prevention. This article explored the decisions taken by the Pharmacy Board of Sierra Leone through its Expert Committee on Medicine Safety and Clinical Trials regarding clinical trials oversight during the Ebola epidemic and the lessons learned. This independent expert committee assessed and provided scientific opinions to the Pharmacy Board of Sierra Leone to inform approval of all clinical trials within 10–15 working days. We also requested for assisted review from the African Vaccine Regulatory Forum and support from the US Food and Drug Administration through a unilateral recognition and reliance memorandum of understanding. In addition, the Agency-ensured structures and systems were in place for reporting and reviewing adverse events and serious adverse events, management of biological samples, submission and review of progress reports, and good clinical practice inspections. Unfortunately, the Ebola epidemic revealed many weaknesses in the country’s clinical trials regulatory structure and processes. Government and partners should further offer more resources to build the clinical trial structures and systems so that the Agency will be better poised to handle future public health emergencies.

The optimum target for systolic and diastolic blood pressure remains divisive. In particular, the conflicting outcomes of the SPRINT and ACCORD trials have led to a divergence of guideline-recommended blood pressure targets for adults with diabetes.Here, we review the existing recommendations for blood pressure targets in diabetes, discussing the evidence base behind them and their limitations. We start by outlining the risks and benefits of lower systolic blood pressure targets among diabetics. We then follow with a separate appraisal of diastolic blood pressure targets, which necessitates examination of the 'J curve' and isolated diastolic hypertension.Current and emerging evidence supports, on balance, a blood pressure therapeutic target of130/90 mmHg in adults at increased risk for cardiovascular disease, including diabetics. Whether certain diabetics with systolic BPs of 120-130 and/or diastolic BPs 80-90 mmHg require drug treatment to a target of120/80 mmHg is less clear and requires more research.

Recent reports have challenged the notion that the lens is immune-privileged. However, these studies have not fully identified the molecular mechanism(s) that promote immune surveillance of the lens. Using a mouse model of targeted glutathione (GSH) deficiency in ocular surface tissues, we have investigated the role of oxidative stress in upregulating cytokine expression and promoting immune surveillance of the eye. RNA-sequencing of lenses from postnatal day (P) 1- aged Gclcf/f;Le-CreTg/− (KO) and Gclcf/f;Le-Cre−/− control (CON) mice revealed upregulation of many cytokines (e.g., CCL4, GDF15, CSF1) and immune response genes in the lenses of KO mice. The eyes of KO mice had a greater number of cells in the aqueous and vitreous humors at P1, P20 and P50 than age-matched CON and Gclcw/w;Le-CreTg/− (CRE) mice. Histological analyses revealed the presence of innate immune cells (i.e., macrophages, leukocytes) in ocular structures of the KO mice. At P20, the expression of cytokines and ROS content was higher in the lenses of KO mice than in those from age-matched CRE and CON mice, suggesting that oxidative stress may induce cytokine expression. In vitro administration of the oxidant, hydrogen peroxide, and the depletion of GSH (using buthionine sulfoximine (BSO)) in 21EM15 lens epithelial cells induced cytokine expression, an effect that was prevented by co-treatment of the cells with N-acetyl-L-cysteine (NAC), a antioxidant. The in vivo and ex vivo induction of cytokine expression by oxidative stress was associated with the expression of markers of epithelial-to-mesenchymal transition (EMT), α-SMA, in lens cells. Given that EMT of lens epithelial cells causes posterior capsule opacification (PCO), we propose that oxidative stress induces cytokine expression, EMT and the development of PCO in a positive feedback loop. Collectively these data indicate that oxidative stress induces inflammation of lens cells which promotes immune surveillance of ocular structures.HighlightsImmune surveillance of ocular structures occurs in mouse eyes deficient in glutathione.Oxidative stress upregulates the expression of pro-inflammatory cytokines (e.g., GDF15, CSF1) in lens cells in vitro and in vivo.The upregulation of cytokines in lens cells is associated with markers of an epithelial-to-mesenchymal transition phenotype.Oxidative stress-induced inflammation and associated epithelial-to-mesenchymal transition may play a role in the development of posterior capsule opacification.

Phenol is an important petrochemical that is conventionally used as a precursor for synthesizing an array of plastics and fine chemicals. As an emerging alternative to its traditional petrochemical production, multiple enzyme pathways have been engineered to date to enable its renewable biosynthesis from biomass feedstocks, each incorporating unique enzyme chemistries and intermediate molecules. Leveraging all three of the unique phenol biosynthesis pathways reported to date, a series of synthetic ‘metabolic funnels’ was engineered, each with the goal of maximizing net precursor assimilation and flux towards phenol via the parallel co-expression of multiple distinct pathways within the same Escherichia coli host. By constructing and evaluating all possible binary and tertiary pathway combinations, one ‘funnel’ was ultimately identified, which supported enhanced phenol production relative to all three individual pathways by 16 to 69%. Further host engineering to increase endogenous precursor availability then allowed for 26% greater phenol production, reaching a final titer of 554 ± 19 mg/L and 28.8 ± 0.34 mg/g yield on glucose. Lastly, using a diphasic culture including dibutyl phthalate for in situ phenol extraction, final titers were further increased to a maximum of 812 ± 145 mg/L at a yield of 40.6 ± 7.2 mg/g. The demonstrated ‘funneling’ pathway holds similar promise in support of phenol production by other, non-E. coli hosts, while this general approach can be readily extended towards a diversity of other value-added bioproducts of interest.

This chapter deals with the area of human rights. It first considers the former position on human rights in the UK, then the European Convention on Human Rights and its incorporation into domestic law by the Human Rights Act 1998. The chapter explores the extent to which human rights law applies to private bodies performing public functions, how it affects the interpretation of legislation, when courts may find legislation compatible with Convention rights and when they may issue a declaration of incompatibility, and examines the adoption and the application of the proportionality test.

This chapter examines the institution of the ombudsman, its origins, the conditions of access and the jurisdiction of the parliamentary, health service and local government ombudsmen and the key ideas of maladministration and injustice. It also reviews how ombudsmen handle and resolve complaints and seek improvements by public bodies in their handling of complaints, and the learning of lessons, as well as considering the outcome of investigations and remedies recommended by the ombudsman. Finally, the chapter discusses the process developed by the ombudsmen in dealing with complainants dissatisfied with the service they have received and how the courts deal with the judicial reviews brought against ombudsmen by complainants. Note is also made of the research on judicial reviews brought against these ombudsmen.

This chapter focuses on devolution. The nature of the asymmetrical devolution of legislative and executive power to Scotland, Wales, and Northern Ireland is outlined, along with the position in England, with the ongoing change to these arrangements analysed. Some issues related to devolution which have been raised in the UK courts are then considered, with the chapter using a range of examples in Scotland, Wales and Northern Ireland to discuss the extent to which the devolution settlement is dynamic and continuing to change. It concludes by exploring the impact of Brexit on devolution in particular.

This chapter discusses the constitutional principle of the rule of law. First the concept of the rule of law is introduced, including discussion and criticisms of Dicey’s understanding of the principle. The difference between formal and substantive understandings of the rule of law is explored, then the chapter examines the rule of law as a broad political doctrine requiring law to be clear and prospective. Finally, the chapter uses in depth analysis of case law to explain the idea of government according to law in the UK: it examines the need for legal authority for official acts, the principle of legality, and the rule of law as a substantive constitutional principle.

This chapter discusses law-making. First, it considers the different types of legislative measures, including the special arrangements introduced for law-making in preparation for Brexit. It then examines the methods of control and influence used before and during the consideration of legislation by Parliament, including analysis of the recently abandoned experiment with ‘English Votes for English Laws’. It concludes by analysing the process of judicial review of delegated legislation.

This chapter examines the operation of the UK’s parliamentary style of government, focusing on the role of Parliament in scrutinising the government. The chapter explains the crucial constitutional convention of ministerial responsibility to Parliament, using examples to demonstrate how it operates in practice. Having set out the principles, the chapter explores the practice of parliamentary scrutiny of government, including the role of select committees, debates and questions on the floor of the Commons, and the role of correspondence between MPs, their constituents, and the government. The chapter concludes by reviewing some reforms intended to re-balance power within Parliament, by strengthening the role of backbench MPs, creating a public petitions system to select topics for debate, and extending parliamentary oversight over public appointments.

This chapter deals with the availability of judicial review and its significance in the constitution. First, it considers the claim for judicial review and the exclusivity principle. It determines who can apply for judicial review and against whom and in respect of what activities judicial review may be sought. Next, it examines the discretionary nature of the remedies available in judicial review proceedings, including how the courts exercise this discretion. The chapter concludes with an examination of the courts’ response to legislative attempts to exclude or oust judicial review.

This chapter introduces the idea of a constitution, analyses its various definitions and characteristics, and explores the specific nature of the UK’s constitutional arrangements. It then examines the key concepts of legitimacy, democracy, legal and political constitutionalism, the conditioning of power (via the principles of the separation of powers and responsible government), and the state.

Cases & Materials on Constitutional and Administrative Law is an invaluable resource. Extracts have been chosen from a wide range of historical and contemporary cases and materials to illustrate the reasoning processes of the courts and to show how legal principles are developed. The extracts from the leading cases in the field are combined with legal, political, and philosophical materials and linked together with explanatory text, alongside extensive notes and questions for discussion. The book takes a critical look at the main doctrines of constitutional law as well as the principles of administrative law, examining the operation of the constitution in relation to Parliament, the government, and the citizen. Incisive commentary throughout the text provides explanation and analysis of the key issues and challenges in constitutional and administrative law. The thirteenth edition has been fully revised and updated to reflect the latest developments in legislation, case law, and politics, including the process and implications of exiting the EU, and the UK’s new post-Brexit legal arrangements; continuing change and challenges to the devolution settlement in Scotland, Wales and Northern Ireland; major Supreme Court decisions in Miller (No.2) / Cherry, UNISON, the Scottish Continuity Bill Reference, and Privacy International; new developments in relation to ministerial responsibility and parliamentary accountability (including the impact of the coronavirus pandemic); proposed repeal of the Fixed-term Parliaments Act 2011; and discussion of proposals for reforms of judicial review and tribunal appeal processes, as well as proposed reform of ombudsmen. This text continues to provide instant access to an unrivalled collection of up-to-date judgments, statutory provisions, official publications, and other policy materials.

This chapter deals with the doctrine of the legislative sovereignty of Parliament. First, the nature of parliamentary sovereignty is considered. Next, the question of whether Parliament can limit the powers of its successors is explored, addressing possible substantive and procedural limits on legislative sovereignty. The chapter then assesses a range of modern challenges to parliamentary sovereignty: those posed by membership of and exit from the EU (including the process and impact of Brexit), human rights, the decision of the House of Lords in Jackson, common law rights and principles, and referendums.

This chapter examines the rationale for giving the task of resolving disputes to statutory tribunals rather than courts. It also describes the structure and organization for most tribunals and how they conduct dispute resolution adjudication. The hearing technique of redress is considered alongside administrative review, particularly the use of mandatory reconsideration in social security to illustrate the advantages and disadvantages of different methods of dispute resolution. Their place in a staged approach, proportionate dispute resolution, is outlined and the possible benefit of conceiving administrative justice as a system with a focus on users is raised as well as some of the issues raised for users by the adoption of digitalization. An outline is given of the oversight activities conducted by the non-statutory Administrative Justice Council.

This chapter examines the notion of judicial independence. It discusses the Constitutional Reform Act 2005 and its provisions reforming the office of the Lord Chancellor, establishing a new Supreme Court, and restructuring judicial appointments. Judicial diversity and discipline, along with further change to the judicial appointments process, are also considered. The chapter also considers the accountability of the judiciary to Parliament and the public, and the relationship between judicial independence and parliamentary privilege.

This chapter reviews the royal prerogative and constitutional conventions, and the relationship between these two sources of constitutional rules. The first section identifies the various types of prerogative power and explores recent examples where these such powers have been placed on a statutory basis, as well as proposals to reverse this process, such as by repealing the Fixed-term Parliaments Act 2011 and reviving the royal prerogative. It also examines attempts to codify constitutional practice, including the Crown’s personal prerogative of the appointment of the Prime Minister in the Cabinet Manual, and the interaction between prerogative and statute in the courts. The second section of the chapter explores constitutional conventions as a source of the constitution, their relationship with law, and their nature as rules of political behaviour. It considers the treatment of conventions in the courts, whether they can obtain legal force, and the feasibility and desirability of codifying conventions. The important connections between the royal prerogative and constitutional conventions are analysed at various points throughout the chapter.