Insights into RNA biology from an atlas of mammalian mRNA-binding proteins.

RNA-binding proteins (RBPs) determine RNA fate from synthesis to decay. Employing two complementary protocols for covalent UV crosslinking of RBPs to RNA, we describe a systematic, unbiased, and comprehensive approach, termed "interactome capture," to define the mRNA interactome of proliferating human HeLa cells. We identify 860 proteins that qualify as RBPs by biochemical and statistical criteria, adding more than 300 RBPs to those previously known and shedding light on RBPs in disease, RNA-binding enzymes of intermediary metabolism, RNA-binding kinases, and RNA-binding architectures. Unexpectedly, we find that many proteins of the HeLa mRNA interactome are highly intrinsically disordered and enriched in short repetitive amino acid motifs. Interactome capture is broadly applicable to study mRNA interactome composition and dynamics in varied biological settings.

Single cell sequencing data identify distinct B cell and fibroblast populations in stricturing Crohn's disease.

Single cell RNA sequencing of human full thickness Crohn's disease (CD) small bowel resection specimens was used to identify potential therapeutic targets for stricturing (S) CD. Using an unbiased approach, 16 cell lineages were assigned within 14,539 sequenced cells from patient-matched SCD and non-stricturing (NSCD) preparations. SCD and NSCD contained identical cell types. Amongst immune cells, B cells and plasma cells were selectively increased in SCD samples. B cell subsets suggested formation of tertiary lymphoid tissue in SCD and compared with NSCD there was an increase in IgG, and a decrease in IgA plasma cells, consistent with their potential role in CD fibrosis. Two Lumican-positive fibroblast subtypes were identified and subclassified based on expression of selectively enriched genes as fibroblast clusters (C) 12 and C9. Cells within these clusters expressed the profibrotic genes Decorin (C12) and JUN (C9). C9 cells expressed ACTA2; ECM genes COL4A1, COL4A2, COL15A1, COL6A3, COL18A1 and ADAMDEC1; LAMB1 and GREM1. GO and KEGG Biological terms showed extracellular matrix and stricture organization associated with C12 and C9, and regulation of WNT pathway genes with C9. Trajectory and differential gene analysis of C12 and C9 identified four sub-clusters. Intra sub-cluster gene analysis detected 13 co-regulated gene modules that aligned along predicted pseudotime trajectories. CXCL14 and ADAMDEC1 were key markers in module 1. Our findings support further investigation of fibroblast heterogeneity and interactions with local and circulating immune cells at earlier time points in fibrosis progression. Breaking these interactions by targeting one or other population may improve therapeutic management for SCD.

Chinese hamster ovary cell line engineering strategies for modular production of custom extracellular vesicles.

Continuously secreted by all cell types, extracellular vesicles (EVs) are small membrane-bound structures which shuttle bioactive cargo between cells across their external environment. Their central role as natural molecular messengers and ability to cross biological barriers has garnered significant attention in the use of EVs as therapeutic delivery vehicles. Still, harnessing the potential of EVs is faced with many obstacles. A cell line engineering approach can be used to exploit EVs to encapsulate a bespoke cargo of interest. However, full details regarding native EV-loading mechanisms remain under debate, making this a challenge. While Chinese hamster ovary (CHO) cells are well known to be the preferred host for recombinant therapeutic protein production, their application as an EV producer cell host has been largely overlooked. In this study, we engineered CHO DG44 cells to produce custom EVs with bespoke cargo. To this end, genetic constructs employing split green fluorescent protein technology were designed for tagging both CD81 and protein cargoes to enable EV loading via self-assembling activity. To demonstrate this, NanoLuc and mCherry were used as model reporter cargoes to validate engineered loading into EVs. Experimental findings indicated that our custom EV approach produced vesicles with up to 15-fold greater cargo compared with commonly used passive loading strategies. When applied to recipient cells, we observed a dose-dependent increase in cargo activity, suggesting successful delivery of engineered cargo via our custom CHO EVs.

Myhre syndrome is caused by dominant-negative dysregulation of SMAD4 and other co-factors.

Myhre syndrome is a connective tissue disorder characterized by congenital cardiovascular, craniofacial, respiratory, skeletal, and cutaneous anomalies as well as intellectual disability and progressive fibrosis. It is caused by germline variants in the transcriptional co-regulator SMAD4 that localize at two positions within the SMAD4 protein, I500 and R496, with I500 V/T/M variants more commonly identified in individuals with Myhre syndrome. Here we assess the functional impact of SMAD4-I500V variant, identified in two previously unpublished individuals with Myhre syndrome, and provide novel insights into the molecular mechanism of SMAD4-I500V dysfunction. We show that SMAD4-I500V can dimerize, but its transcriptional activity is severely compromised. Our data show that SMAD4-I500V acts dominant-negatively on SMAD4 and on receptor-regulated SMADs, affecting transcription of target genes. Furthermore, SMAD4-I500V impacts the transcription and function of crucial developmental transcription regulator, NKX2-5. Overall, our data reveal a dominant-negative model of disease for SMAD4-I500V where the function of SMAD4 encoded on the remaining allele, and of co-factors, are perturbed by the continued heterodimerization of the variant, leading to dysregulation of TGF and BMP signaling. Our findings not only provide novel insights into the mechanism of Myhre syndrome pathogenesis but also extend the current knowledge of how pathogenic variants in SMAD proteins cause disease.

Standardised method for cardiomyocyte isolation and purification from individual murine neonatal, infant, and adult hearts.

Primary cardiomyocytes are invaluable for understanding postnatal heart development. However, a universal method to obtain freshly purified cardiomyocytes without using different age-dependent isolation procedures and cell culture, is lacking. Here, we report the development of a standardised method that allows rapid isolation and purification of high-quality cardiomyocytes from individual neonatal through to adult C57BL/6J murine hearts. Langendorff retrograde perfusion, which is currently limited to adult hearts, was adapted for use in neonatal and infant hearts by developing an easier in situ aortic cannulation technique. Tissue digestion conditions were optimised to achieve efficient digestion of hearts of all ages in a comparable timeframe (<14 min). This resulted in a high yield (1.56-2.2 × 106 cells/heart) and viability (~70-100%) of cardiomyocytes post-isolation. An immunomagnetic cell separation step was then applied to yield highly purified cardiomyocytes (~95%) as confirmed by immunocytochemistry, flow cytometry, and qRT-PCR. For cell type-specific studies, cardiomyocyte DNA, RNA, and protein could be extracted in sufficient yields to conduct molecular experiments. We generated transcriptomic datasets for neonatal cardiomyocytes from individual hearts, for the first time, which revealed nine sex-specific genes (FDR < 0.05) encoded on the sex chromosomes. Finally, we also developed an in situ fixation protocol that preserved the native cytoarchitecture of cardiomyocytes (~94% rod-shaped post-isolation), and used it to evaluate cell morphology during cardiomyocyte maturation, as well as capture spindle-shaped neonatal cells undergoing cytokinesis. Together, these procedures allow molecular and morphological profiling of high-quality cardiomyocytes from individual hearts of any postnatal age.

Ride-Hailing and Road Traffic Crashes: A Critical Review.

Ride-hailing businesses, including Uber and Lyft, have reshaped road traffic since they first began operating in the United States approximately a decade ago. It follows that ride-hailing may also alter the incidence and distribution of road traffic crash injuries and deaths. The available evidence relating ride-hailing to crashes is critically reviewed in this article. We present a theoretical model that synthesizes the hypothesized mechanisms, and we identify common methodological challenges and suggest priorities for future research. Mixed results have been reported for the overall incidence of road traffic crash injuries and deaths, likely due to heterogeneous impacts on vehicular traffic flow (e.g., increasing the volume of vehicles); on vehicle-, person-, and event-level characteristics (e.g., reducing alcohol-impaired driver crashes); on road-user types (e.g., increasing pedestrian crashes); and on environmental conditions (e.g., reducing crashes most substantially where public transit access is poorest). The lack of a well-developed theory of human mobility and methodological challenges that are common to many ecological studies impede exploration of these sources of moderation. Innovative solutions are required to explicate ride-hailing's heterogeneous impacts, to guide policy that can take advantage of the public health benefits of ride-hailing, and to ensure that research keeps pace with technological advances that continue to reshape road traffic use.

Heterozygous loss of WBP11 function causes multiple congenital defects in humans and mice.

The genetic causes of multiple congenital anomalies are incompletely understood. Here, we report novel heterozygous predicted loss-of-function (LoF) and predicted damaging missense variants in the WW domain binding protein 11 (WBP11) gene in seven unrelated families with a variety of overlapping congenital malformations, including cardiac, vertebral, tracheo-esophageal, renal and limb defects. WBP11 encodes a component of the spliceosome with the ability to activate pre-messenger RNA splicing. We generated a Wbp11 null allele in mouse using CRISPR-Cas9 targeting. Wbp11 homozygous null embryos die prior to E8.5, indicating that Wbp11 is essential for development. Fewer Wbp11 heterozygous null mice are found than expected due to embryonic and postnatal death. Importantly, Wbp11 heterozygous null mice are small and exhibit defects in axial skeleton, kidneys and esophagus, similar to the affected individuals, supporting the role of WBP11 haploinsufficiency in the development of congenital malformations in humans. LoF WBP11 variants should be considered as a possible cause of VACTERL association as well as isolated Klippel-Feil syndrome, renal agenesis or esophageal atresia.

Trends in inpatient care for psychiatric disorders in NHS hospitals across England, 1998/99-2019/20: an observational time series analysis.

PURPOSE: It is unclear how hospitals are responding to the mental health needs of the population in England, against a backdrop of diminishing resources. We aimed to document patterns in hospital activity by psychiatric disorder and how these have changed over the last 22 years. METHODS: In this observational time series analysis, we used routinely collected data on all NHS hospitals in England from 1998/99 to 2019/20. Trends in hospital admissions and bed days for psychiatric disorders were smoothed using negative binomial regression models with year as the exposure and rates (per 1000 person-years) as the outcome. When linear trends were not appropriate, we fitted segmented negative binomial regression models with one change-point. We stratified by gender and age group [children (0-14 years); adults (15 years +)]. RESULTS: Hospital admission rates and bed days for all psychiatric disorders decreased by 28.4 and 38.3%, respectively. Trends were not uniform across psychiatric disorders or age groups. Admission rates mainly decreased over time, except for anxiety and eating disorders which doubled over the 22-year period, significantly increasing by 2.9% (AAPC = 2.88; 95% CI: 2.61-3.16; p < 0.001) and 3.4% (AAPC = 3.44; 95% CI: 3.04-3.85; p < 0.001) each year. Inpatient hospital activity among children showed more increasing and pronounced trends than adults, including an increase of 212.9% for depression, despite a 63.8% reduction for adults with depression during the same period. CONCLUSION: In the last 22 years, there have been overall reductions in hospital activity for psychiatric disorders. However, some disorders showed pronounced increases, pointing to areas of growing need for inpatient psychiatric care, especially among children.

Effectiveness of cash-plus programmes on early childhood outcomes compared to cash transfers alone: A systematic review and meta-analysis in low- and middle-income countries.

BACKGROUND: To strengthen the impact of cash transfers, these interventions have begun to be packaged as cash-plus programmes, combining cash with additional transfers, interventions, or services. The intervention's complementary ("plus") components aim to improve cash transfer effectiveness by targeting mediating outcomes or the availability of supplies or services. This study examined whether cash-plus interventions for infants and children <5 are more effective than cash alone in improving health and well-being. METHODS AND FINDINGS: Forty-two databases, donor agencies, grey literature sources, and trial registries were systematically searched, yielding 5,097 unique articles (as of 06 April 2021). Randomised and quasi-experimental studies were eligible for inclusion if the intervention package aimed to improve outcomes for children <5 in low- and middle-income countries (LMICs) and combined a cash transfer with an intervention targeted to Sustainable Development Goal (SDG) 2 (No Hunger), SDG3 (Good Health and Well-being), SDG4 (Education), or SDG16 (Violence Prevention), had at least one group receiving cash-only, examined outcomes related to child-focused SDGs, and was published in English. Risk of bias was appraised using Cochrane Risk of Bias and ROBINS-I Tools. Random effects meta-analyses were conducted for a cash-plus intervention category when there were at least 3 trials with the same outcome. The review was preregistered with PROSPERO (CRD42018108017). Seventeen studies were included in the review and 11 meta-analysed. Most interventions operated during the first 1,000 days of the child's life and were conducted in communities facing high rates of poverty and often, food insecurity. Evidence was found for 10 LMICs, where most researchers used randomised, longitudinal study designs (n = 14). Five intervention categories were identified, combining cash with nutrition behaviour change communication (BCC, n = 7), food transfers (n = 3), primary healthcare (n = 2), psychosocial stimulation (n = 7), and child protection (n = 4) interventions. Comparing cash-plus to cash alone, meta-analysis results suggest Cash + Food Transfers are more effective in improving height-for-age (d = 0.08 SD (0.03, 0.14), p = 0.02) with significantly reduced odds of stunting (OR = 0.82 (0.74, 0.92), p = 0.01), but had no added impact in improving weight-for-height (d = -0.13 (-0.42, 0.16), p = 0.24) or weight-for-age z-scores (d = -0.06 (-0.28, 0.15), p = 0.43). There was no added impact above cash alone from Cash + Nutrition BCC on anthropometrics; Cash + Psychosocial Stimulation on cognitive development; or Cash + Child Protection on parental use of violent discipline or exclusive positive parenting. Narrative synthesis evidence suggests that compared to cash alone, Cash + Primary Healthcare may have greater impacts in reducing mortality and Cash + Food Transfers in preventing acute malnutrition in crisis contexts. The main limitations of this review are the few numbers of studies that compared cash-plus interventions against cash alone and the potentially high heterogeneity between study findings. CONCLUSIONS: In this study, we observed that few cash-plus combinations were more effective than cash transfers alone. Cash combined with food transfers and primary healthcare show the greatest signs of added effectiveness. More research is needed on when and how cash-plus combinations are more effective than cash alone, and work in this field must ensure that these interventions improve outcomes among the most vulnerable children.

Synthetic Control Methods for the Evaluation of Single-Unit Interventions in Epidemiology: A Tutorial.

Evaluating the impacts of population-level interventions (e.g., changes to state legislation) can be challenging as conducting randomized experiments is often impractical and inappropriate, especially in settings where the intervention is implemented in a single, aggregate unit (e.g., a country or state). A common nonrandomized alternative is to compare outcomes in the treated unit(s) with unexposed controls both before and after the intervention. However, the validity of these designs depends on the use of controls that closely resemble the treated unit on before-intervention characteristics and trends on the outcome, and suitable controls may be difficult to find because the number of potential control regions is typically limited. The synthetic control method provides a potential solution to these problems by using a data-driven algorithm to identify an optimal weighted control unit-a "synthetic control"-based on data from before the intervention from available control units. While popular in the social sciences, the method has not garnered as much attention in health research, perhaps due to a lack of accessible texts aimed at health researchers. We address this gap by providing a comprehensive, nontechnical tutorial on the synthetic control method, using a worked example evaluating Florida's "stand your ground" law to illustrate methodological and practical considerations.

Firearm Homicide Incidence, Within-state Firearm Laws, and Interstate Firearm Laws in US Counties.

BACKGROUND: Firearm homicides occur less frequently in US states with more firearm control laws. However, firearms are easily transported across state lines, and laws in one location may affect firearm violence in another. This study examined associations between within-state firearm laws and firearm homicide while accounting for interference from laws in other nearby states. METHODS: The units of analysis were 3,107 counties in the 48 contiguous US states, arrayed in 15 yearly panels for 2000 to 2014 (n = 46,605). The dependent measure was firearm homicides accessed from the Centers for Disease Control and Prevention (CDC) Compressed Mortality Data. The main independent measures were counts of firearm laws and the proportion of laws within categories (e.g., background checks, child access prevention laws). We calculated these measures for interstate laws using a geographic gravity function between county centroids. Bayesian conditional autoregressive Poisson models related within-state firearm laws and interstate firearm laws to firearm homicides. RESULTS: There were 172,726 firearm homicides in the included counties over the 15 years. States had between 3 and 100 firearm laws. Within-state firearm laws (incidence rate ratio [IRR] = 0.995, 95% confidence interval [CI] = 0.992, 0.997) and interstate firearm laws (IRR = 0.993, 95% CI = 0.990, 0.996) were independently associated with fewer firearm homicides, and associations for within-state laws were strongest where interstate laws were weakest. CONCLUSIONS: Additional firearm laws are associated with fewer firearm homicides both within the states where the laws are enacted and elsewhere in the United States. Interference from interstate firearm laws may bias associations for studies of within-state laws and firearm homicide.

A comparative analysis of recombinant Fab and full-length antibody production in Chinese hamster ovary cells.

Monoclonal antibodies are the leading class of biopharmaceuticals in terms of numbers approved for therapeutic purposes. Antigen-binding fragments (Fab) are also used as biotherapeutics and used widely in research applications. The dominant expression systems for full-length antibodies are mammalian cell-based, whereas for Fab molecules the preference has been an expression in bacterial systems. However, advances in CHO and downstream technologies make mammalian systems an equally viable option for small- and large-scale Fab production. Using a panel of full-length IgG antibodies and their corresponding Fab pair with different antigen specificities, we investigated the impact of the IgG and Fab molecule format on production from Chinese hamster ovary (CHO) cells and assessed the cellular capability to process and produce these formats. The full-length antibody format resulted in the recovery of fewer mini-pools posttransfection when compared to the corresponding Fab fragment format that could be interpreted as indicative of a greater overall burden on cells. Antibody-producing cell pools that did recover were subsequently able to achieve higher volumetric protein yields (mg/L) and specific productivity than the corresponding Fab pools. Importantly, when the actual molecules produced per cell of a given format was considered (as opposed to mass), CHO cells produced a greater number of Fab molecules per cell than obtained with the corresponding IgG, suggesting that cells were more efficient at making the smaller Fab molecule. Analysis of cell pools showed that gene copy number was not correlated to the subsequent protein production. The amount of mRNA correlated with secreted Fab production but not IgG, whereby posttranscriptional processes act to limit antibody production. In summary, we provide the first comparative description of how full-length IgG and Fab antibody formats impact on the outcomes of a cell line construction process and identify potential limitations in their production that could be targeted for engineering increases in the efficiency in the manufacture of these recombinant antibody formats.

Effects of Laws Expanding Civilian Rights to Use Deadly Force in Self-Defense on Violence and Crime: A Systematic Review.

Background. Since 2005, most US states have expanded civilian rights to use deadly force in self-defense outside the home. In most cases, legislation has included removing the duty to retreat anywhere one may legally be, commonly known as stand-your-ground laws. The extent to which these laws affect public health and safety is widely debated in public and policy discourse.Objectives. To synthesize the available evidence on the impacts and social inequities associated with changing civilian rights to use deadly force in self-defense on violence, injury, crime, and firearm-related outcomes.Search Methods. We searched MEDLINE, Embase, PsycINFO, Scopus, Web of Science, Sociological Abstracts, National Criminal Justice Reference Service Abstracts, Education Resources Information Center, International Bibliography of the Social Sciences, ProQuest Dissertations and Theses, Google Scholar, National Bureau of Economic Research working papers, and SocArXiv; harvested references of included studies; and consulted with experts to identify studies until April 2020.Selection Criteria. Eligible studies quantitatively estimated the association between laws that expanded or restricted the right to use deadly force in self-defense and population or subgroup outcomes among civilians with a comparator.Data Collection and Analysis. Two reviewers extracted study data using a common form. We assessed study quality using the Risk of Bias in Nonrandomized Studies of Interventions tools adapted for (controlled) before-after studies. To account for data dependencies, we conducted graphical syntheses (forest plots and harvest plots) to summarize the evidence on impacts and inequities associated with changing self-defense laws.Main Results. We identified 25 studies that estimated population-level impacts of laws expanding civilian rights to use deadly force in self-defense, all of which focused on stand-your-ground or other expansions to self-defense laws in the United States. Studies were scored as having serious or critical risk of bias attributable to confounding. Risk of bias was low across most other domains (i.e., selection, missing data, outcome, and reporting biases). Stand-your-ground laws were associated with no change to small increases in violent crime (total and firearm homicide, aggravated assault, robbery) on average across states. Florida-based studies showed robust increases (24% to 45%) in firearm and total homicide while self-defense claims under stand-your-ground law were more often denied when victims were White, especially when claimants were racial minorities.Author's Conclusions. The existing evidence contradicts claims that expanding self-defense laws deters violent crime across the United States. In at least some contexts, including Florida, stand-your-ground laws are associated with increases in violence, and there are racial inequities in the application of these laws.Public Health Implications. In some US states, most notably Florida, stand-your-ground laws may have harmed public health and safety and exacerbated social inequities. Our findings highlight the need for scientific evidence on both population and equity impacts of self-defense laws to guide legislative action that promotes public health and safety for all.Trial Registration. Open Science Framework (https://osf.io/uz68e).

Ridesharing and motor vehicle crashes: a spatial ecological case-crossover study of trip-level data.

BACKGROUND: Ridesharing services (eg, Uber, Lyft) have facilitated over 11 billion trips worldwide since operations began in 2010, but the impacts of ridesharing on motor vehicle injury crashes are largely unknown. - METHODS: This spatial ecological case-cross over used highly spatially and temporally resolved trip-level rideshare data and incident-level injury crash data for New York City (NYC) for 2017 and 2018. The space-time units of analysis were NYC taxi zone polygons partitioned into hours. For each taxi zone-hour we calculated counts of rideshare trip origins and rideshare trip destinations. Case units were taxi zone-hours in which any motor vehicle injury crash occurred, and matched control units were the same taxi zone from 1 week before (-168 hours) and 1 week after (+168 hours) the case unit. Conditional logistic regression models estimated the odds of observing a crash (separated into all injury crashes, motorist injury crashes, pedestrian injury crashes, cyclist injury crashes) relative to rideshare trip counts. Models controlled for taxi trips and other theoretically relevant covariates (eg, precipitation, holidays). RESULTS: Each additional 100 rideshare trips originating within a taxi zone-hour was associated with 4.6% increased odds of observing any injury crash compared with the control taxi zone-hours (OR=1.046; 95% CI 1.032 to 1.060). Associations were detected for motorist injury and pedestrian injury crashes, but not cyclist injury crashes. Findings were substantively similar for analyses conducted using trip destinations as the exposure of interest. CONCLUSIONS: Ridesharing contributes to increased injury burden due to motor vehicle crashes, particularly for motorist and pedestrian injury crashes at trip nodes.

Ularcirc: visualization and enhanced analysis of circular RNAs via back and canonical forward splicing.

Circular RNAs (circRNA) are a unique class of transcripts that can only be identified from sequence alignments spanning discordant junctions, commonly referred to as backsplice junctions (BSJ). Canonical splicing is also linked with circRNA biogenesis either from the parental transcript or internal to the circRNA, and is not fully utilized in circRNA software. Here we present Ularcirc, a software tool that integrates the visualization of both BSJ and forward splicing junctions and provides downstream analysis of selected circRNA candidates. Ularcirc utilizes the output of CIRI, circExplorer, or raw chimeric output of the STAR aligner and assembles BSJ count table to allow multi-sample analysis. We used Ularcirc to identify and characterize circRNA from public and in-house generated data sets and demonstrate how it can be used to (i) discover novel splicing patterns of parental transcripts, (ii) detect internal splicing patterns of circRNA, and (iii) reveal the complexity of BSJ formation. Furthermore, we identify circRNA that have potential open reading frames longer than their linear sequence. Finally, we detected and validated the presence of a novel class of circRNA generated from ApoA4 transcripts whose BSJ derive from multiple non-canonical splicing sites within coding exons. Ularcirc is accessed via https://github.com/VCCRI/Ularcirc.

The impact of equity factors on receipt of timely appropriate care for children with suspected malaria in eastern Uganda.

INTRODUCTION: Malaria accounts for more than one-tenth of sub-Saharan Africa's 2.8 million annual childhood deaths, and remains a leading cause of post-neonatal child mortality in Uganda. Despite increased community-based treatment in Uganda, children continue to die because services fail to reach those most at risk. This study explores the influence of two key equity factors, socioeconomic position and rurality, on whether children with fever in eastern Uganda receive timely access to appropriate treatment for suspected malaria. METHODS: This was a cross-sectional study in which data were collected from 1094 caregivers of children aged 6-59 months on: illness and care-seeking during the previous two weeks, treatment received, and treatment dosing schedule. Additional data on rurality and household socioeconomic position were extracted from the Iganga-Mayuge Health and Demographic Surveillance Site (HDSS) database. A child was considered to have received prompt and appropriate care for symptoms of malaria if they received the recommended drug in the recommended dosing schedule on the day of symptom onset or the next day. Unadjusted and adjusted logistic regression models were developed to explore associations of the two equity factors with the outcome. The STROBE checklist for observational studies guided reporting. RESULTS: Seventy-four percent of children had symptoms of illness in the preceding two weeks, of which fever was the most common. Children from rural households were statistically more likely to receive prompt and appropriate treatment with artemisinin-combination therapy than their semi-urban counterparts (OR 2.32, CI 1.17-4.59, p = 0.016). This association remained significant following application of an adjusted regression model that included the age of the child, caregiver relationship, and household wealth index (OR 2.4, p = 0.036). Wealth index in its own right did not exert a significant effect for children with reported fever (OR for wealthiest quintile = 1.02, CI 0.48-2.15, p = 0.958). CONCLUSIONS: The findings from this study help to identify the role and importance of two key equity determinants on care seeking and treatment receipt for fever in children. Whilst results should be interpreted within the limitations of data and context, further studies have the potential to assist policy makers to target inequitable social and spatial variations in health outcomes as a key strategy in ending preventable child morbidity and mortality.

Dissecting the mechanism of action of intravenous immunoglobulin in human autoimmune disease: Lessons from therapeutic modalities targeting Fcγ receptors.

Since the first description of the administration of high doses of pooled serum IgG, also referred to as intravenous IgG (IVIg) therapy, as being able to ameliorate various autoimmune diseases, researchers have been investigating which molecular and cellular pathways underlie IVIg activity. Apart from trying to understand the obvious conundrum that IgG can trigger both autoimmune pathology and resolution of inflammation, the rapidly expanding use of IVIg has led to a lack of availability of this primary blood product, providing a strong rationale for developing recombinant alternatives. During the last decade, a tremendous number of novel insights into IVIg activity brought the goal of replacing IVIg within reach, at least in select indications, and has led to the initiation of several clinical trials. At the forefront of this effort is the modulation of autoantibody half-life and blocking access of autoantibodies to fragment cystallizable γ receptors (Fcγ receptors). In this rostrum article, we will briefly discuss current models of IVIg activity, followed by a more specific focus on novel therapeutic avenues that are entering the clinic and may replace IVIg in the future.

NAD Deficiency, Congenital Malformations, and Niacin Supplementation.

BACKGROUND: Congenital malformations can be manifested as combinations of phenotypes that co-occur more often than expected by chance. In many such cases, it has proved difficult to identify a genetic cause. We sought the genetic cause of cardiac, vertebral, and renal defects, among others, in unrelated patients. METHODS: We used genomic sequencing to identify potentially pathogenic gene variants in families in which a person had multiple congenital malformations. We tested the function of the variant by using assays of in vitro enzyme activity and by quantifying metabolites in patient plasma. We engineered mouse models with similar variants using the CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 system. RESULTS: Variants were identified in two genes that encode enzymes of the kynurenine pathway, 3-hydroxyanthranilic acid 3,4-dioxygenase (HAAO) and kynureninase (KYNU). Three patients carried homozygous variants predicting loss-of-function changes in the HAAO or KYNU proteins (HAAO p.D162*, HAAO p.W186*, or KYNU p.V57Efs*21). Another patient carried heterozygous KYNU variants (p.Y156* and p.F349Kfs*4). The mutant enzymes had greatly reduced activity in vitro. Nicotinamide adenine dinucleotide (NAD) is synthesized de novo from tryptophan through the kynurenine pathway. The patients had reduced levels of circulating NAD. Defects similar to those in the patients developed in the embryos of Haao-null or Kynu-null mice owing to NAD deficiency. In null mice, the prevention of NAD deficiency during gestation averted defects. CONCLUSIONS: Disruption of NAD synthesis caused a deficiency of NAD and congenital malformations in humans and mice. Niacin supplementation during gestation prevented the malformations in mice. (Funded by the National Health and Medical Research Council of Australia and others.).

Long-term Exposure to Neighborhood Deprivation and Intimate Partner Violence Among Women: A UK Birth Cohort Study.

BACKGROUND: Intimate partner violence is the most common form of violence perpetrated against women. To our knowledge, the effect of neighborhood disadvantage on intimate partner violence against women has never been investigated prospectively outside the United States. METHODS: We used data from the Avon Longitudinal Study of Parents and Children in the United Kingdom, which followed our target sample, 7,219 women, from birth and their mothers (from pregnancy). At age 21, 2,128 participants self-reported the frequency of experiencing physical, psychological, or sexual intimate partner violence since age 18. Participants' exposure to neighborhood-level deprivation and family-level socioeconomic characteristics (e.g., income) were measured at 10 time points from baseline (gestation) until children were 18 years old. We estimated the effect of cumulative exposure to greater neighborhood-level deprivation on the risk of experiencing intimate partner violence using marginal structural models with stabilized inverse probability weights, accounting for time-varying confounding by socioeconomic indicators and sample attrition. RESULTS: A one-unit increase in cumulative exposure to more severe neighborhood deprivation was associated with a 62% increase in participants' frequency of experiencing intimate partner violence (95% confidence interval 11%, 237%) and 36% increase in their risk of experiencing any intimate partner violence (95% confidence interval 1%, 85%). CONCLUSIONS: In our study, cumulative exposure to greater neighborhood deprivation over the first 18 years of life was associated with women's increased risk of experiencing intimate partner violence in early adulthood. Future studies should test this association across contexts, including underlying mechanisms, and evaluate preventive strategies that target structural disparities.

Evaluating the impact of penalising the use of mobile phones while driving on road traffic fatalities, serious injuries and mobile phone use: a systematic review.

BACKGROUND: A vast literature has demonstrated that using mobile phones while driving increases the risk of road traffic crashes. In response, policy-makers have introduced bans and harsher penalties on using mobile phones while driving. Even though emerging evidence suggests that such measures may reduce mobile phone use and crashes, the literature has not been systematically reviewed and synthesised. OBJECTIVE: To evaluate the impact of penalising mobile phone use while driving on road traffic fatalities, serious injuries and the prevalence of mobile phone use while driving. METHODS: We employed a comprehensive search strategy using electronic databases, websites, handsearching and other sources to locate studies evaluating legislation on mobile phone use while driving. Randomised controlled trials, interrupted time series', controlled before-after studies with control(s) not exposed to harsher sanctions and panel data designs were included if they measured the outcomes of fatalities, serious injuries or the prevalence of mobile phone use while driving. Eligible studies were critically appraised. Due to substantial heterogeneity, the results were synthesised narratively. The synthesis structured studies according to the type of legislation and outcome measure. RESULTS: Of the 7420 studies retrieved, 32 were included. The evidence on the effects of penalising mobile phone use while driving was weak, and somewhat inconsistent, but pointed to a potential decrease in the prevalence of mobile phone use and fatalities for all-driver primary enforcement hand-held bans and texting bans. CONCLUSIONS: Preventing fatalities from risky driving practices may be helped by implementing harsher laws that penalise mobile phone use while driving.