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Changing climate and land‐use practices have the potential to bring previously isolated populations of pest insects into new sympatry. This heightens the need to better understand how differing patterns of host–plant association, and unique endosymbionts, serve to promote genetic isolation or integration. We addressed these factors in populations of potato psyllid, Bactericera cockerelli (Šulc), a generalist herbivore that vectors a bacterial pathogen (Candidatus Liberibacter solanacearum, causal pathogen of zebra chip disease) of potato (Solanum tuberosum L.). Genome‐wide SNP data revealed two major genetic clusters—psyllids collected from potato crops were genetically similar to psyllids found on a common weed, Lycium spp., but dissimilar from those found on another common non‐crop host, Solanum dulcamara L. Most psyllids found on Lycium spp. and potato represented a single mitochondrial cytochrome oxidase I (COI) haplotype that has been suggested to not be native to the region, and whose arrival may have been concurrent with zebra chip disease first emerging. The putatively introduced COI haplotype usually co‐occurred with endosymbiotic Wolbachia, while the putatively resident COI haplotype generally did not. Genetic intermediates between the two genetic populations of insects were rare, consistent with recent sympatry or reproductive isolation, although admixture patterns of apparent hybrids were consistent with introgression of genes from introduced into resident populations. Our results suggest that both host–plant associations and endosymbionts are shaping the population genetic structure of sympatric psyllid populations associated with different non‐crop hosts. It is of future interest to explicitly examine vectorial capacity of the two populations and their potential hybrids, as population structure and hybridization might alter regional vector capacity and disease outbreaks.

Detecting a tropospheric ozone trend from sparsely sampled ozonesonde profiles (typically once per week) is challenging due to the short-lived anomalies in the time series resulting from ozone's high temporal variability. To enhance trend detection, we have developed a sophisticated statistical approach that utilizes a geoadditive model to assess ozone variability across a time series of vertical profiles. Treating the profile time series as a set of individual time series on discrete pressure surfaces, a class of smoothing spline ANOVA (analysis of variance) models is used for the purpose of jointly modeling multiple correlated time series (on separate pressure surfaces) by their associated seasonal and interannual variabilities. This integrated fit method filters out the unstructured variation through a statistical regularization (i.e., a roughness penalty) by taking advantage of the additional correlated data points available on the pressure surfaces above and below the surface of interest. We have applied this technique to the trend analysis of the vertically correlated time series of tropospheric ozone observations from (1) IAGOS (In-service Aircraft for a Global Observing System) commercial aircraft profiles above Europe and China throughout 1994–2017 and (2) NOAA GML's (Global Monitoring Laboratory) ozonesonde records at Hilo, Hawaii, (1982–2018) and Trinidad Head, California (1998–2018). We illustrate the ability of this technique to detect a consistent trend estimate and its effectiveness in reducing the associated uncertainty in the profile data due to the low sampling frequency. We also conducted a sensitivity analysis of frequent IAGOS profiles above Europe (approximately 120 profiles per month) to determine how many profiles in a month are required for reliable long-term trend detection. When ignoring the vertical correlation, we found that a typical sampling strategy (i.e. four profiles per month) might result in 7 % of sampled trends falling outside the 2σ uncertainty interval derived from the full dataset with an associated 10 % of mean absolute percentage error. Based on a series of sensitivity studies, we determined optimal sampling frequencies for (1) basic trend detection and (2) accurate quantification of the trend. When applying the integrated fit method, we find that a typical sampling frequency of four profiles per month is adequate for basic trend detection; however, accurate quantification of the trend requires 14 profiles per month. Accurate trend quantification can be achieved with only 10 profiles per month if a regular sampling frequency is applied. In contrast, the standard separated fit method, which ignores the vertical correlation between pressure surfaces, requires 8 profiles per month for basic trend detection and 18 profiles per month for accurate trend quantification. While our method improves trend detection from sparse datasets, the key to substantially reducing the uncertainty is to increase the sampling frequency.

Although nonprofit collaboration is commonplace, recent research suggests that faith-based organizations (FBOs) are less likely to collaborate than other nonprofits. This study builds on prior FBO, collaboration, and nonprofit capacity research to examine the influence of religiosity and operational capacity on FBOs’ within- and cross-sector partnerships. Findings from a survey with 197 FBOs across the United States reveal a complex picture of how religiosity and operational capacity influence FBO collaboration. More specifically, staff religiosity was positively related to cross-sector partnerships. Service religiosity (i.e., religious elements in staff–client interactions) was negatively associated with collaboration with government agencies. Results also indicated that FBOs with higher operational capacity had more partners in the nonprofit, business, and public sectors. These findings suggest that FBOs generally lack the operational capacity for collaboration and that service religiosity creates additional barriers to it. This article concludes with implications for research in FBOs, interorganizational collaboration, and nonprofit capacity.

Murine studies have shown the potential for γδ T cells to mediate immunity to Staphylococcus aureus in multiple tissue settings by the secretion of diverse cytokines. However, the role played by γδ T cells in human immune responses to S. aureus is almost entirely unknown. In this study, we establish the capacity of human Vδ2+ γδ T cells for rapid activation in response to S. aureus. In coculture with S. aureus–infected monocyte-derived dendritic cells (DCs), Vδ2+ cells derived from peripheral blood rapidly upregulate CD69 and secrete high levels of IFN-γ. DCs mediate this response through direct contact and IL-12 secretion. In turn, IFN-γ released by Vδ2+ cells upregulates IL-12 secretion by DCs in a positive feedback loop. Furthermore, coculture with γδ T cells results in heightened expression of the costimulatory molecule CD86 and the lymph node homing molecule CCR7 on S. aureus–infected DCs. In cocultures of CD4+ T cells with S. aureus–infected DCs, the addition of γδ T cells results in heightened CD4+ T cell activation. Our findings identify γδ T cells as potential key players in the early host response to S. aureus during bloodstream infection, promoting enhanced responses by both innate and adaptive immune cell populations, and support their consideration in the development of host-directed anti–S. aureus treatments.

The literature on sleep and eating disorder (ED) diagnoses is scarce, particularly so in the developmental period of adolescence. Despite a dearth of published findings, it appears that the majority of individuals with EDs suffer from disturbed sleep. There are some inconsistencies in the current literature; however, only a few studies have assessed sleep in this population utilizing objective measures. In this article, we provide an overview of the current available research and suggest an agenda for future directions to better understand the sleep-ED association and corresponding treatment implications. Research that includes objective and subjective measures to assess sleep problems, stratified by ED diagnosis, and in a treatment-specific context is necessary to inform future prevention and intervention efforts.

A generic method to estimate the relative feasibility of formation of high-entropy compounds in a single phase, directly from first principles, is developed. As a first step, the relative formation...

Objectives To evaluate the efficacy of two novel compounds against mycobacteria and determine the molecular basis of their action on DNA gyrase using structural and mechanistic approaches. Methods Redx03863 and Redx04739 were tested in antibacterial assays, and also against their target, DNA gyrase, using DNA supercoiling and ATPase assays. X-ray crystallography was used to determine the structure of the gyrase B protein ATPase sub-domain from Mycobacterium smegmatis complexed with the aminocoumarin drug novobiocin, and structures of the same domain from Mycobacterium thermoresistibile complexed with novobiocin, and also with Redx03863. Results Both compounds, Redx03863 and Redx04739, were active against selected Gram-positive and Gram-negative species, with Redx03863 being the more potent, and Redx04739 showing selectivity against M. smegmatis. Both compounds were potent inhibitors of the supercoiling and ATPase reactions of DNA gyrase, but did not appreciably affect the ATP-independent relaxation reaction. The structure of Redx03863 bound to the gyrase B protein ATPase sub-domain from M. thermoresistibile shows that it binds at a site adjacent to the ATP- and novobiocin-binding sites. We found that most of the mutations that we made in the Redx03863-binding pocket, based on the structure, rendered gyrase inactive. Conclusions Redx03863 and Redx04739 inhibit gyrase by preventing the binding of ATP. The fact that the Redx03863-binding pocket is distinct from that of novobiocin, coupled with the lack of activity of resistant mutants, suggests that such compounds could have potential to be further exploited as antibiotics.

The spatial pattern of flea (Siphonaptera: Pulicidae) infestation risk in cats and dogs across Great Britain is quantified, using data collected from a national survey undertaken in 2018, with particular attention given to the association between insecticidal treatment and infestation risk. Flea infestation risk declined significantly from south to north. None of the factors: pet breed, sex, neutered status, or whether the pet had been abroad, showed any relationship with the underlying geographic distribution, which is most likely to be associated with climatic factors. However, overall, only 23.6% of the cats and 35% of the dogs inspected had been treated with identifiable flea products that were still ‘in date’ at the point of inspection. The percentage of owners treating their pet broadly followed infestation risk. The insecticide fipronil is a common active in a wide range of flea treatments and was the most frequently applied insecticide class, particularly in cats. However, 62% of cats and 45% of dogs that had been treated with a fipronil-based product that was ‘in date’ at the point of inspection still had fleas. Persistent flea infestation is likely to be due to a range of factors, including compliance and application failure, but the data provide strong inferential evidence for a lack of efficacy of fipronil-based products. Given the ubiquity of flea infestation, this finding and the relatively low-level of treatment compliance, highlight a clear need for greater owner education about the importance of flea management and a better understanding of the efficacy of different products.

Introduction Inflammasomes are multiprotein complexes that regulate immune processes in response to infections and tissue damage. They modulate Interleukin-1beta (IL-1β) expression, a major proinflammatory cytokine. The inflammasome/IL-1β pathway is involved in head and neck squamous cell carcinoma (HNSCC) progression and the periodontal pathogens Fusobacterium nucleatum (Fn) and Porphyromonas gingivalis (Pg) have been reported to cause chronic inflammation in HNSCC. The aim of this study was to characterise the role of these pathogens in regulating inflammasome activity and the IL-1β response in HNSCC in vitro. Methods An HNSCC cell line (H400) was exposed to Fn and Pg individually or in combination for 24h, ± incubation for 30 min with 5 mM adenosine triphosphate (ATP). Transcript levels of inflammasomes, NLRP3 and AIM2; inflammasome-regulatory proteins, POP1, CARD16 and TRIM16; and inflammasome-component, ASC and caspase 1 and IL-1β, were assayed by RT-PCR. Expression of IL-1β was by immunocytochemistry and ELISA. Results NLRP3 expression was significantly upregulated in response to Pg, Fn + Pg, Pg + ATP and Fn + Pg + ATP. AIM2 was significantly upregulated by Fn, Pg and Fn + Pg + ATP exposure. All conditions significantly upregulated IL-1β gene expression. POP1 expression was significantly downregulated by Pg or Fn exposure but not by Fn + Pg. Intracellular pro- and mature IL-1β were significantly higher following Fn and Pg + ATP exposure. Conclusion Pg alone increased IL-1β by upregulating AIM2, NLRP3 and downregulating POP1. Fn promoted IL-1β by increasing AIM2 and downregulating POP1. Pg + ATP with or without Fn upregulated NLRP3, IL-1β by downregulating POP1. Periodontal pathogens may contribute to HNSCC pathogenesis by increasing the IL-1β response due to inflammasome dysregulation.

ContextInterscholastic heat policies for football have not been evidence based. Therefore, their effectiveness in mitigating exertional heat illness has not been assessed.ObjectiveTo discuss the development of the Georgia High School Association heat policy and assess the effectiveness of revised guidelines.DesignDescriptive epidemiology study.SettingGeorgia high schools.Patients or Other ParticipantsInterscholastic football players in grades 9 through 12.Main Outcome Measure(s)Heat syncope and heat exhaustion (HS/HE) illness rates (IRs) were calculated per 1000 athlete-exposures (AEs), and relative risk (RR) was calculated as a ratio of postpolicy (POST) IR divided by prepolicy (PRE) IR.ResultsA total of 214 HS/HE cases (172 PRE, 42 POST) and 341 348 AEs (178 230 PRE, 163 118 POST) were identified. During the first 5 days of the PRE period, approximately 50% of HS/HE illnesses occurred; HS/HE IRs doubled when practice sessions increased from 2 to 2.5 hours and tripled for practices ≥3 hours. The HS/HE IRs in the PRE period increased from 0.44/1000 AEs for wet-bulb globe temperatures (WBGTs) of 2.0/1000 AEs for WBGTs from 87°F (30.6°C) to 89.9°F (32.2°C). The RRs comparing PRE and POST policy periods were 0.29 for WBGTs of 90°F (>32.3°C).ConclusionsResults from the PRE period guided the Georgia High School Association to revise its heat and humidity policy to include a mandated 5-day acclimatization period when no practices may exceed 2 hours and the use of WBGT-based activity-modification categories. The new policy reduced HS/HE IRs by 35% to 100%, depending on the WBGT category. Our results may be generalizable to other states with hot and humid climates similar to that of Georgia.

Female steelhead trout (Oncorhynchus mykiss) reared for 1 year (traditional, “S1”) and 2 years (experimental, “S2”) in captivity in the Methow River Basin, Washington, produced very similar numbers of offspring in replicate spawning channels over 3 consecutive years. In contrast, S1 anadromous males outcompeted S2 males for access to nesting females and, as a result, produced significantly more offspring. Male dominance was positively associated with body mass, but body mass did not account for differences between S1 and S2 males. Much smaller precocious S2 males that matured during rearing in hatchery raceways sired offspring in all six breeding groups. Contributions from precocious males were nearly always the result of sneak spawning events, although on rare occasions, precocious males were able to stimulate females to spawn in the absence of an anadromous male. Similarities in female breeding success suggest that S1 and S2 hatchery steelhead should exhibit similar productivity under natural conditions, but S1 male steelhead may result in greater rates of gene flow from hatchery to natural populations.

Every day, hundreds of millions of people worldwide take nonsteroidal anti-inflammatory drugs (NSAIDs), often in conjunction with multiple other medications. In the bloodstream, NSAIDs are mostly bound to serum albumin (SA). We report the crystal structures of equine serum albumin complexed with four NSAIDs (ibuprofen, ketoprofen, etodolac, and nabumetone) and the active metabolite of nabumetone (6-methoxy-2-naphthylacetic acid, 6-MNA). These compounds bind to seven drug-binding sites on SA. These sites are generally well-conserved between equine and human SAs, but ibuprofen binds to both SAs in two drug-binding sites, only one of which is common. We also compare the binding of ketoprofen by equine SA to binding of it by bovine and leporine SAs. Our comparative analysis of known SA complexes with FDA-approved drugs clearly shows that multiple medications compete for the same binding sites, indicating possibilities for undesirable physiological effects caused by drug–drug displacement or competition with common metabolites. We discuss the consequences of NSAID binding to SA in a broader scientific and medical context, particularly regarding achieving desired therapeutic effects based on an individual’s drug regimen.

Aim: This study examined the direct and indirect associations between childhood psychopathology symptoms, screen use, media multitasking and participation in non-digital recreation. Methods: Psychopathology symptoms, media use, media multitasking, participation in sports, social clubs and reading/games were reported by 520 parents about their 3- to 11-year-old children. The data were analysed using structural equation modelling. Results: There were bidirectional negative associations between sports participation and emotional problems (β = −0.16, P

Pediatric human immunodeficiency virus post-exposure prophylaxis is frequently indicated, but delays in medication receipt are common. Using plan-do-study-act cycles, we developed a multidisciplinary collaboration to reduce critical process delays in our pediatric emergency department. Interruptions decreased from a median 1 per month pre-intervention to zero per month during the intervention.

Polistes paper wasps in the Fuscopolistes subgenus (Hymenoptera: Vespidae) can be serious pests when they swarm at tall man-made structures. Chemical attractants may be useful to trap such paper wasps when they achieve pest status. Polistes venom has been shown to elicit a variety of behavioral responses in congeneric wasps, making it a source for potential chemical attractants. The compound N-(3-methylbutyl)acetamide is a principal volatile component in the venom of many female vespid wasps, including numerous Polistes species. We report the presence of N-(3-methylbutyl)acetamide in autumn gynes of Polistes metricus Say, Polistes bellicosus Cresson, and Polistes dorsalis (F.), as well as workers of Polistes aurifer (Saussure), P. bellicosus, P. metricus, and P. dorsalis. In field tests conducted in Florida, Georgia, South Carolina, and Washington, N-(3-methylbutyl)acetamide attracted male and female P. aurifer and P. metricus, as well as male P. dorsalis and P. bellicosus. Thus, N-(3-methylbutyl)acetamide may be a useful lure for trapping these paper wasps in pest situations.

Aim\ud \ud To establish if irrigant activation techniques, namely manual‐dynamic‐activation (MDA), passive‐ultrasonic‐irrigation (PUI) and sonic‐irrigation (SI), improve the tubular penetration of sodium hypochlorite (NaOCl) into root dentine when compared with conventional‐needle‐irrigation (CNI). Secondly, investigate if increasing NaOCl concentration and/or contact‐time improves the performance of these techniques.\ud Methodology\ud \ud A total of 83 extracted human maxillary permanent canines were decoronated to 15 mm and root canals prepared to a size 40,.10 taper. Root dentine was stained with crystal violet for 72 h and embedded in silicone. Eighty specimens were randomly distributed into 16 groups (n = 5) according to the irrigant activation technique, NaOCl concentration (2%; 5.25%) and irrigant contact‐time (10 min; 20 min). All activation techniques were used for 60 s in the last minute of irrigation. Additionally, 3 teeth were not exposed to NaOCl to confirm adequate dentine staining had occurred (i.e. negative control). All specimens were subsequently dissected, observed under a light microscope and NaOCl penetration depth (µm) determined by measuring the average width of bleached dentine using ImageJ software. Statistical comparisons were made with paired and unpaired t‐tests, ANOVAs followed by post‐hoc Tukey and Dunnett’s tests, and a general linear model (α < 0.05).\ud Results\ud \ud Overall, NaOCl penetration ranged from 38.8 µm – 411.0 µm with MDA, PUI and SI consistently resulting in significantly greater tubular infiltration than CNI (P < 0.05). The deepest measurements in the coronal, middle and apical segments were all recorded in the MDA; 5.25%; 20 min group and the least in the CNI; 2%; 10 min group. Increasing either irrigant concentration or contact‐time resulted in significantly greater NaOCl penetration depths for all techniques and segments of the canal (P < 0.05). However, when irrigant concentration and contact‐time were increased together, a significant interaction effect between these two independent variables was observed on overall NaOCl penetration (P < 0.05).\ud Conclusions\ud \ud Agitating irrigants with MDA, PUI or SI, as well as using greater irrigant concentrations or contact‐times, potentiated NaOCl penetration into root dentine. However, longer durations of NaOCl exposure at lower concentrations resulted in similar depths of tubular penetration as those achieved at higher concentrations.

This article reports on a multiple-case study of how six educational alliances developed Integrated Logic Models, a new tool that integrates components of multiple programs into a single logic mode...

This symposium presents three studies that apply Bridging Multiple Worlds and other complementary theories to analyze how students and families engage their cultural resources to create multiple pa...

Bed-load transport plays a critical role in river morphological change and has an important impact on river ecology. Although there is good understanding of the role of the variation of river bed grain size on transport dynamics in equilibrium conditions, much less is understood for non-equilibrium conditions when the channel is either aggrading or degrading. In particular, the relative role of different grain sizes in the promotion and hindering of the transport of coarse and fine fractions in a degrading channel has yet to be investigated. The current study attempts to provide new understanding through a series of flume experiments done using uniform and graded sediment particles. The experiments revealed coarser grain-size fractions for a poorly-sorted sediment, relative to uniform-sized sediment, reduced the transport of finer grains and finer fractions enhanced the transport of coarse grains. This hindering-promotion effect, caused by relative hiding and exposure of finer and coarse fractions, increased with bed slope and decreased with relative submergence. In particular, as relative submergence increased, the graded fractions tended towards behaving more like their uniform-sized counterparts. Also, the bed-load parameter of the graded fractions increased more with a rise in bed slope than observed for the uniform-sized counterparts. These results revealed, for degrading channel conditions, such as downstream of a dam, bed-load equations developed for uniform bed sediment are inappropriate for use in natural river systems, particularly in mountain streams. Furthermore, changes in river bed composition due to activities that enhance the input of hill-slope sediment, such as fire, logging, and agricultural development, are likely to cause significant changes in river morphology.

Background: The role of nutrient timing in energy intake amongst people with dysglycaemia is understudied but could be a simple method to help regulate appetite. This study analysed within-person associations of sugar intake at breakfast and subsequent energy intake. Methods: We used 4-day diet diary data from 147 participants (47 % men) encompassing 547 days of diet recording in the Sedentary Time and Metabolic Health in People with Type 2 diabetes project (STAMP-2). Linear two-level models were used to investigate within- (day-level) and between-person (participant-level) variation in total and post-breakfast energy intake according to skipping breakfast, low- (> 0-14.2 g) or high- (> 14.2 g) sugar intake at breakfast, adjusting for potential confounding or mediation. Results: Post-breakfast energy intakes were observed to be lower after eating low- and high-sugar breakfasts (compared to skipping), but higher total energy intake was associated with eating a high-sugar breakfast. Compared to breakfast skipping, both low- and high-sugar breakfasts were strongly associated with lower post-breakfast energy intake (-178, 95 % confidence interval [CI] -261, -94 kcal/d; ‑151, 95 % CI -235, -67 kcal/d, respectively). However, compared to skipping breakfast, low-sugar breakfasts were weakly associated with higher total daily energy intake (64, 95 % CI -18, 146 kcal/d), whereas high-sugar breakfasts had a strong association (135, 95 % CI 52, 217 kcal/d). Post-breakfast energy intakes were similar between low- and high-sugar breakfast days (27, 95 % CI -53, 106 kcal/d), whilst total daily energy intake tended to be higher with high- (compared to low-) sugar breakfasts (70, -8, 149 kcal/d). We also observed evidence of energy compensation whereby 86 % of the extra energy consumed in a low-sugar breakfast was compensated for by reductions in post-breakfast energy intakes, compared to only 53 % after a high-sugar breakfast. Conclusion: Overall, high-, but not low-, sugar breakfasts were associated with higher total daily energy intake when compared to breakfast skipping, despite a similar reduction in post-breakfast energy intake. We found evidence of poorer post-breakfast energy intake compensation with high-sugar breakfasts than low-sugar breakfasts. These findings suggest portion size may be important to consider in future breakfast research, with sugar being a proxy of portion size.

Background Vaccination of children and young people against SARS-CoV-2 is recommended in some countries. Scarce data have been published on immune responses induced by COVID-19 vaccines in people younger than 18 years compared with the same data that are available in adults. Methods COV006 is a phase 2, single-blind, randomised, controlled trial of ChAdOx1 nCoV-19 (AZD1222) in children and adolescents at four trial sites in the UK. Healthy participants aged 6–17 years, who did not have a history of chronic respiratory conditions, laboratory-confirmed COVID-19, or previously received capsular group B meningococcal vaccine (the control), were randomly assigned to four groups (4:1:4:1) to receive two intramuscular doses of 5 × 1010 viral particles of ChAdOx1 nCoV-19 or control, 28 days or 84 days apart. Participants, clinical investigators, and the laboratory team were masked to treatment allocation. Study groups were stratified by age, and participants aged 12–17 years were enrolled before those aged 6–11 years. Due to the restrictions in the use of ChAdOx1 nCoV-19 in people younger than 30 years that were introduced during the study, only participants aged 12–17 years who were randomly assigned to the 28-day interval group had received their vaccinations at the intended interval (day 28). The remaining participants received their second dose at day 112. The primary outcome was assessment of safety and tolerability in the safety population, which included all participants who received at least one dose of the study drug. The secondary outcome was immunogenicity, which was assessed in participants who were seronegative to the nucleocapsid protein at baseline and received both prime and boost vaccine. This study is registered with ISRCTN (15638344). Findings Between Feb 15 and April 2, 2021, 262 participants (150 [57%] participants aged 12–17 years and 112 [43%] aged 6–11 years; due to the change in the UK vaccination policy, the study terminated recruitment of the younger age group before the planned number of participants had been enrolled) were randomly assigned to receive vaccination with two doses of either ChAdOx1 nCoV-19 (n=211 [n=105 at day 28 and n=106 at day 84]) or control (n=51 [n=26 at day 28 and n=25 at day 84]). One participant in the ChAdOx1 nCoV-19 day 28 group in the younger age bracket withdrew their consent before receiving a first dose. Of the participants who received ChAdOx1 nCoV-19, 169 (80%) of 210 participants reported at least one solicited local or systemic adverse event up to 7 days following the first dose, and 146 (76%) of 193 participants following the second dose. No serious adverse events related to ChAdOx1 nCoV-19 administration were recorded by the data cutoff date on Oct 28, 2021. Of the participants who received at least one dose of ChAdOx1 nCoV-19, there were 128 unsolicited adverse events up to 28 days after vaccination reported by 83 (40%) of 210 participants. One participant aged 6–11 years receiving ChAdOx1 nCoV-19 reported a grade 4 fever of 40·2°C on day 1 following first vaccination, which resolved within 24 h. Pain and tenderness were the most common local solicited adverse events for all the ChAdOx1 nCoV-19 and capsular group B meningococcal groups following both doses. Of the 242 participants with available serostatus data, 14 (6%) were seropositive at baseline. Serostatus data were not available for 20 (8%) of 262 participants. Among seronegative participants who received ChAdOx1 nCoV-19, anti-SARS-CoV-2 IgG and pseudoneutralising antibody titres at day 28 after the second dose were higher in participants aged 12–17 years with a longer interval between doses (geometric means of 73 371 arbitrary units [AU]/mL [95% CI 58 685–91 733] and 299 half-maximal inhibitory concentration [IC50; 95% CI 230–390]) compared with those aged 12–17 years who received their vaccines 28 days apart (43 280 AU/mL [95% CI 35 852–52 246] and 150 IC50 [95% CI 116–194]). Humoral responses were higher in those aged 6–11 years than in those aged 12–17 years receiving their second dose at the same 112-day interval (geometric mean ratios 1·48 [95% CI 1·07–2·07] for anti-SARS-CoV-2 IgG and 2·96 [1·89–4·62] for pseudoneutralising antibody titres). Cellular responses peaked after a first dose of ChAdOx1 nCoV-19 across all age and interval groups and remained above baseline after a second vaccination. Interpretation ChAdOx1 nCoV-19 is well tolerated and immunogenic in children aged 6–17 years, inducing concentrations of antibody that are similar to those associated with high efficacy in phase 3 studies in adults. No safety concerns were raised in this trial. Funding AstraZeneca and the UK Department of Health and Social Care through the UK National Institute for Health and Care Research.

BackgroundWalk-in clinics are non-hospital based primary care facilities that are designed to operate without appointments and provide increased healthcare access with extended hours. Urgent Primary Care Centres (UPCCs) were introduced to British Columbia (BC) in 2018 as an additional primary care resource that provides urgent, but not emergent care during extended hours. This study identifies publicly-reported access characteristics for episodic primary care in BC, and provides a clinic-level comparison between walk-in clinics and UPCCs.MethodologyThis cross-sectional study used publicly available data from all walk-in clinics and UPCCs in BC. A structured data collection form was used to record access characteristics from clinic websites, including business hours, weekend availability, attachment to a longitudinal family practice, and provision of virtual services.ResultsIn total, 268 clinics were included in the analysis (243 walk-in clinics, 25 UPCCs). Of those, 225 walk-in clinics (92.6%) and two UPCCs (8.0%) were attached to a longitudinal family practice. Only 153 (63%) walk-in clinics offered weekend services, compared to 24 (96%) of UPCCs. Walk-in clinics offer the majority (8,968.6/ 78.4%) of their service hours between 08:00 and 17:00, Monday to Friday. UPCCs offer the majority (889.3/ 53.7%) of their service hours after 17:00.ConclusionMost walk-in clinics are associated with a longitudinal family practice and provide the majority of clinic services during typical business hours. More research that includes patient characteristics and care outcomes, analyzed at the clinic level, may be useful to support the optimization of episodic primary health care delivery.

BACKGROUND Advances in medical treatments in recent years have contributed to an overall decline in HIV-related opportunistic infections and deaths in youth; however, mortality and morbidity rates in perinatally and non-perinatally infected youth living with HIV (YLWH) remain relatively high today. OBJECTIVE The goal of this project is to assess the usage, utility, and cost-effectiveness of PlusCare, a mobile/web application (app) for HIV case management for YLWH. The app supports routine case management tasks such as scheduling follow-up visits, sharing documents for review and signature, lab test results, and between-visit communications (e.g., encouraging messages). METHODS We conducted a single-group mixed-methods, pre-post study conducted with HIV case management programs in 2 large, urban hospitals in the Boston metro area. Case management staff (CMs, N=20) and YLWH patients (N=45) participated in the study with access to PlusCare for up to 15 months and 12 months, respectively. RESULTS CMs and YLWH reported System Usability Scale scores of 51 (SD=7.9) and 63 (SD=10.6), respectively. Although marginally significant, total charges billed at one of the two sites compared to the 12 months prior to app use (including emergency, inpatient, and outpatient charges) decreased by 41% (P=.046). We also observed slight increases in YLWH self-reported self-efficacy (SEMCD6) and quality of life (CDC HRQOL-4) from baseline to 12-month follow-up (P=.02 and P=.03, respectively) and increases in self-efficacy from 6-month to 12-month follow-up (P=.02). There was no significant change in HIV viral suppression or appointment or medication adherence in this small sample pilot study. CONCLUSIONS While perceived usability was low, qualitative feedback from CMs and usage patterns suggest direct messaging and timely, remote, and secure sharing of lab results and documents (including electronic signatures) between CMs and YLWH can be particularly useful and have potential value in supporting care coordination and promotion of patient self-efficacy and quality of life. CLINICALTRIAL ClinicalTrials.gov NCT03758066

Background: Packed red blood cells (PRBC) are often ordered for pediatric cardiac catheterizations (PCC) with no current appropriate use guidelines, resulting in increased utilization. A quality improvement (QI) project was created to focus on optimizing ordering of PRBC for PCC. The primary aim was to increase adherence to new guidelines to > 97%, with a global aim to decrease patient cost. Methods: Retrospective chart review was performed to collect baseline data. For PDSA cycle 1, we updated our own PRBC ordering guidelines by risk stratifying procedures. The PCC report was updated to include documentation of PRBC ordering or transfusion. PDSA cycle 2 included further refinement of the guidelines, and improved awareness amongst those placing the pre-PCC orders. Process measures included ensuring the need for PRBC stated during every pre-PCC time-out, and consistent documentation in the PCC report. Balancing measures included monitoring the rate of PRBC transfusion within 24 hours post-PCC, and incidence of emergency release of PRBCs. Results: For 3 months of baseline data, 112/403 of CCPs (27.8%) had PRBC ordered, 87.5% (98/112) of which followed the prior protocol, with 13/403 (3.2%) receiving PRBC transfusion during CCP. With our revised guidelines, there could be > 20% reduction in PRBC ordering, with estimated > $20,000 cost savings based on 1 crossmatch ($173) and 1 unit PRBC ($730) prepared. For PDSA cycle 1, 27/81 (33%) of PCC had PRBC ordered, with 92% adherence to the updated guidelines, and 100% documentation compliance. For PDSA cycle 2, 13/47 (27.6%) had PRBC ordered with 100% adherence to ordering guidelines. During both cycles, 3 patients had PRBC transfusions within 24 hours of their procedure for other medical reasons not related to bleeding. No cases required emergency release of PRBC. There was a statistically significant process change from a median of 86% to 100% adherence (p=0.01) since implementation of the updated guidelines. Conclusion: With revised guidelines and improved adherence, hospital resources and charges were decreased in low-risk cardiac catheterization procedures without an increase in adverse events. This model can be utilized in other systems to similarly reduce resource utilization and costs.

Metals and metal-based compounds comprise multifarious pharmaco-active and toxicological xenobiotics. From heavy metal toxicity to chemotherapeutics, the toxicokinetics of these compounds have both historical and modern-day relevance. Zebrafish have become an attractive model organism in elucidating pharmaco- and toxicokinetics in environmental exposure and clinical translation studies. Although zebrafish studies have the benefit of being higher-throughput than rodent models, there are several significant constraints to the model. One such limitation is inherent in the waterborne dosing regimen. Water concentrations from these studies cannot be extrapolated to provide reliable internal dosages. Direct measurements of the metal-based compounds allow for a better correlation with compound-related molecular and biological responses. To overcome this limitation for metals and metal-based compounds, a technique was developed to digest zebrafish larval tissue after exposure and quantify metal concentrations within tissue samples by inductively coupled plasma mass spectrometry (ICPMS). ICPMS methods were used to determine the metal concentrations of platinum (Pt) from cisplatin and ruthenium (Ru) from several novel Ru-based chemotherapeutics in zebrafish tissue. Additionally, this protocol distinguished concentrations of Pt that were sequestered in the chorion of the larval compared with the zebrafish tissue. These results indicate that this method can be applied to quantitate the metal dose present in larval tissues. Further, this method may be adjusted to identify specific metals or metal-based compounds in a broad range of exposure and dosing studies.

The objective of this study was to characterize and compare the dry-aging flavor precursors and their liberation mechanisms in beef aged with different methods. Thirteen paired loins were collected at 5 days postmortem, divided into four sections, and randomly assigned into four aging methods (wet-aging (WA), conventional dry-aging (DA), dry-aging in a water-permeable bag (DWA), and UV-light dry-aging (UDA)). All sections were aged for 28 days at 2 °C, 65% RH, and a 0.8 m/s airflow before trimming and sample collection for chemical, metabolomics, and microbiome analyses. Higher concentrations of free amino acids and reducing sugars were observed in all dry-aging samples (p < 0.05). Similarly, metabolomics revealed greater short-chain peptides in the dry-aged beef (p < 0.05). The DWA samples had an increase in polyunsaturated free fatty acids (C18:2trans, C18:3n3, C20:2, and C20:5; p < 0.05) along with higher volatile compound concentrations compared to other aging methods (aldehyde, nonanal, octanal, octanol, and carbon disulfide; p < 0.05). Microbiome profiling identified a clear separation in beta diversity between dry and wet aging methods. The Pseudomonas spp. are the most prominent bacterial species in dry-aged meat, potentially contributing to the greater accumulation of flavor precursor concentrations in addition to the dehydration process during the dry-aging. Minor microbial species involvement, such as Bacillus spp., could potentially liberate unique and potent flavor precursors.

To determine the ability of different irrigation solutions to biomechanically remove Enterococcus faecalis biofilm from a novel artificial root canal model during chemomechanical preparation.High resolution micro-computer-tomography scans of a mandibular molar's mesial root were used to produce 50 identical 3D-printed resin root canal models. These were cultured with E.faecalis over seven days to generate biofilm and subjected to chemomechanical preparation using: saline; 17% ethylenediaminetetraacetic acid (EDTA) or 2% sodium hypochlorite (NaOCl) alongside positive/negative controls (n = 10). Canals were prepared to 40/.06 taper, with 1 mL irrigation between instruments, followed by 5 mL penultimate rinse, 30 s ultrasonic activation and 5 mL final rinse. Residual biofilm volume (pixels) was determined following immunofluorescent staining and confocal-laser-scanning-microscopy imaging. Statistical comparisons were made using Kruskal-Wallis with post-hoc Dunn's tests (α ⟨0.05).In all canal thirds, the greatest biofilm removal was observed with NaOCl, followed by EDTA and saline. The latter had significantly higher E.faecalis counts than NaOCl and EDTA (P ⟨0.01). However, no statistical differences were found between EDTA and NaOCl or saline and positive controls (P ⟩0.05).Within limitations of this model, 17% EDTA was found to be as effective as 2% NaOCl at eradicating E.faecalis biofilm following chemomechanical preparation. Further investigations with multi-species biofilms are encouraged.

Measurements of galaxy rotation curves provide direct measurements of the distribution of baryonic and dark matter in galaxies. Here, we present spectroscopic confirmation and one such rotation curve for a z = 0.5325 galaxy observed with Keck I/MOSFIRE as a filler target for the Web Epoch of Reionization Survey. The rotation curve was derived from Hα 6563 Å emission out to a galactocentric radius of approximately 24 kpc. The target's rotation curve is well fit by an arctangent curve, that when combined with broadbanned photometric constraints on the galaxy’s stellar mass, predicts a dark matter fraction consistent with results from the literature for z ∼ 0.5. We constrain the estimate for this galaxy's dark matter fraction to be 93%, out to a galactocentric radius of 30 kpc.

Within regenerative endodontics, exciting opportunities exist for the development of next-generation targeted biomaterials that harness epigenetic machinery, including microRNAs (miRNAs), histone acetylation, and DNA methylation, which are used to control pulpitis and to stimulate repair. Although histone deacetylase inhibitors (HDACi) and DNA methyltransferase inhibitors (DNMTi) induce mineralisation in dental pulp cell (DPC) populations, their interaction with miRNAs during DPC mineralisation is not known. Here, small RNA sequencing and bioinformatic analysis were used to establish a miRNA expression profile for mineralising DPCs in culture. Additionally, the effects of a HDACi, suberoylanilide hydroxamic acid (SAHA), and a DNMTi, 5-aza-2′-deoxycytidine (5-AZA-CdR), on miRNA expression, as well as DPC mineralisation and proliferation, were analysed. Both inhibitors increased mineralisation. However, they reduced cell growth. Epigenetically-enhanced mineralisation was accompanied by widespread changes in miRNA expression. Bioinformatic analysis identified many differentially expressed mature miRNAs that were suggested to have roles in mineralisation and stem cell differentiation, including regulation of the Wnt and MAPK pathways. Selected candidate miRNAs were demonstrated by qRT-PCR to be differentially regulated at various time points in mineralising DPC cultures treated with SAHA or 5-AZA-CdR. These data validated the RNA sequencing analysis and highlighted an increased and dynamic interaction between miRNA and epigenetic modifiers during the DPC reparative processes.

Numerous studies have demonstrated the normal tissue-sparing effects of ultra-high dose rate ‘FLASH’ irradiation in vivo, with an associated reduction in damage burden being reported in vitro. Towards this, two key radiochemical mechanisms have been proposed: radical–radical recombination (RRR) and transient oxygen depletion (TOD), with both being proposed to lead to reduced levels of induced damage. Previously, we reported that FLASH induces lower levels of DNA strand break damage in whole-blood peripheral blood lymphocytes (WB-PBL) ex vivo, but our study failed to distinguish the mechanism(s) involved. A potential outcome of RRR is the formation of crosslink damage (particularly, if any organic radicals recombine), whilst a possible outcome of TOD is a more anoxic profile of induced damage resulting from FLASH. Therefore, the aim of the current study was to profile FLASH-induced damage via the Comet assay, assessing any DNA crosslink formation as a putative marker of RRR and/or anoxic DNA damage formation as an indicative marker of TOD, to determine the extent to which either mechanism contributes to the “FLASH effect”. Following FLASH irradiation, we see no evidence of any crosslink formation; however, FLASH irradiation induces a more anoxic profile of induced damage, supporting the TOD mechanism. Furthermore, treatment of WB-PBLs pre-irradiation with BSO abrogates the reduced strand break damage burden mediated by FLASH exposures. In summary, we do not see any experimental evidence to support the RRR mechanism contributing to the reduced damage burden induced by FLASH. However, the observation of a greater anoxic profile of damage following FLASH irradiation, together with the BSO abrogation of the reduced strand break damage burden mediated by FLASH, lends further support to TOD being a driver of the reduced damage burden plus a change in the damage profile mediated by FLASH.

A complete census of dusty star-forming galaxies (DSFGs) at early epochs is necessary to constrain the obscured contribution to the cosmic star formation rate density (CSFRD), however DSFGs beyond $z \sim 4$ are both rare and hard to identify from photometric data alone due to degeneracies in submillimeter photometry with redshift. Here, we present a pilot study obtaining follow-up Atacama Large Millimeter Array (ALMA) $2\,$mm observations of a complete sample of 39 $850\,\rm\mu m$-bright dusty galaxies in the SSA22 field. Empirical modeling suggests $2\,$mm imaging of existing samples of DSFGs selected at $850\,\rm\mu m - 1\,$mm can quickly and easily isolate the "needle in a haystack" DSFGs that sit at $z>4$ or beyond. Combining archival submillimeter imaging with our measured ALMA $2\,$mm photometry ($1\sigma \sim 0.08\,$mJy$\,$beam$^{-1}$ rms), we characterize the galaxies' IR SEDs and use them to constrain redshifts. With available redshift constraints fit via the combination of six submillimeter bands, we identify 6/39 high-$z$ candidates each with $>50\%$ likelihood to sit at $z > 4$, and find a positive correlation between redshift and $2\,$mm flux density. Specifically, our models suggest the addition of $2\,$mm to a moderately constrained IR SED will improve the accuracy of a millimeter-derived redshift from $\Delta z/(1+z) = 0.3$ to $\Delta z/(1+z) = 0.2$. Our IR SED characterizations provide evidence for relatively high emissivity spectral indices ($\langle \beta \rangle = 2.4\pm0.3$) in the sample. We measure that especially bright ($S_{850\rm\mu m}>5.55\,$mJy) DSFGs contribute $\sim10$% to the cosmic-averaged CSFRD from $2, Comment: 22 pages, 7 figures, accepted for publication in ApJ

Excessive dietary intake of fat results in its storage in white adipose tissue (WAT). Energy expenditure through lipid oxidation occurs in brown adipose tissue (BAT). Certain WAT depots can undergo a change termed beiging where markers that BAT express are induced. Little is known about signalling pathways inducing beiging. Here, inhibition of a signalling pathway regulating alternative pre-mRNA splicing is involved in adipocyte beiging. Clk1/2/4 kinases regulate splicing by phosphorylating factors that process pre-mRNA. Clk1 inhibition by TG003 results in beige-like adipocytes highly expressing PGC1α and UCP1. SiRNA for Clk1, 2 and 4, demonstrated that Clk1 depletion increased UCP1 and PGC1α expression, whereas Clk2/4 siRNA did not. TG003-treated adipocytes contained fewer lipid droplets, are smaller, and contain more mitochondria, resulting in proton leak increases. Additionally, inhibition of PKCβII activity, a splice variant regulated by Clk1, increased beiging. PGC1α is a substrate for both Clk1 and PKCβII kinases, and we surmised that inhibition of PGC1α phosphorylation resulted in beiging of adipocytes. We show that TG003 binds Clk1 more than Clk2/4 through direct binding, and PGC1α binds to Clk1 at a site close to TG003. Furthermore, we show that TG003 is highly specific for Clk1 across hundreds of kinases in our activity screen. Hence, Clk1 inhibition becomes a target for induction of beige adipocytes.

Transforming growth factor

Human dental pulp stem cells (hDPSCs) have increasingly gained interest as a potential therapy for nerve regeneration in medicine and dentistry, however their neurogenic potential remains a matter of debate. This study aimed to characterize hDPSC neuronal differentiation in comparison with the human SH-SY5Y neuronal stem cell differentiation model. Both hDPSCs and SH-SY5Y could be differentiated to generate typical neuronal-like cells following sequential treatment with all-trans retinoic acid (ATRA) and brain-derived neurotrophic factor (BDNF), as evidenced by significant expression of neuronal proteins βIII-tubulin (TUBB3) and neurofilament medium (NF-M). Both cell types also expressed multiple neural gene markers including growth-associated protein 43 (GAP43), enolase 2/neuron-specific enolase (ENO2/NSE), synapsin I (SYN1), nestin (NES), and peripherin (PRPH), and exhibited measurable voltage-activated Na+ and K+ currents. In hDPSCs, upregulation of acetylcholinesterase (ACHE), choline O-acetyltransferase (CHAT), sodium channel alpha subunit 9 (SCN9A), POU class 4 homeobox 1 (POU4F1/BRN3A) along with a downregulation of motor neuron and pancreas homeobox 1 (MNX1) indicated that differentiation was more guided toward a cholinergic sensory neuronal lineage. Furthermore, the Extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor U0126 significantly impaired hDPSC neuronal differentiation and was associated with reduction of the ERK1/2 phosphorylation. In conclusion, this study demonstrates that extracellular signal-regulated kinase/Mitogen-activated protein kinase (ERK/MAPK) is necessary for sensory cholinergic neuronal differentiation of hDPSCs. hDPSC-derived cholinergic sensory neuronal-like cells represent a novel model and potential source for neuronal regeneration therapies.

Introduction Weak grip strength is associated with a range of adverse health outcomes, and an accelerated decline in grip strength confers an even greater risk. To date there has been limited research into the factors associated with change in grip strength in mid-life. Using data from UK Biobank our aim was to investigate the associations of long-term conditions (LTCs), multimorbidity and lifestyle factors with patterns of change in grip strength. Methods We used data from 44,315 UK Biobank participants with grip strength measured at baseline (2006–10) and a subsequent study visit around nine years later. At baseline, participants’ LTCs were self-reported and categorised against a hierarchy. Multimorbidity was characterised by the number of LTC categories present (0, 1, 2 and 3+). Lifestyle factors (body mass index (BMI), smoking, diet and physical activity) were also assessed. Change in grip strength was grouped into four patterns: decline, stable low, stable high or reference (no change or increase) and used as the outcome in multinomial logistic regression models. Results Most LTC categories were associated with adverse patterns of change in grip strength (stable low and/or decline): for example, musculoskeletal/trauma conditions were associated with an increased risk of the stable low pattern (Relative Risk Ratio [RRR] = 1.63; 95% Confidence Interval [CI] 1.49–1.79). Multimorbidity and lifestyle factors had independent associations with grip strength change: those with 3+ categories of LTCs were more likely to experience decline in grip strength (RRR = 1.18;95% CI 1.08–1.28) compared to those with no LTCs. Low physical activity was associated with adverse patterns of grip strength, while raised BMI had divergent associations (both the decline and stable high patterns of grip strength change). Conclusion Individuals living with multimorbidity and those with lifestyle risk factors such as low physical activity are at increased risk of loss of strength over time.

Iron surface determinant B (IsdB) is a hemoglobin (Hb) receptor essential for hemic iron acquisition by Staphylococcus aureus. Heme transfer to IsdB is possible from oxidized Hb (metHb), but inefficient from Hb either bound to oxygen (oxyHb) or bound to carbon monoxide (HbCO), and encompasses a sequence of structural events that are currently poorly understood. By single-particle cryo-electron microscopy, we determined the structure of two IsdB:Hb complexes, representing key species along the heme extraction pathway. The IsdB:HbCO structure, at 2.9-Å resolution, provides a snapshot of the preextraction complex. In this early stage of IsdB:Hb interaction, the hemophore binds to the β-subunits of the Hb tetramer, exploiting a folding-upon-binding mechanism that is likely triggered by a cis/trans isomerization of Pro173. Binding of IsdB to α-subunits occurs upon dissociation of the Hb tetramer into α/β dimers. The structure of the IsdB:metHb complex reveals the final step of the extraction process, where heme transfer to IsdB is completed. The stability of the complex, both before and after heme transfer from Hb to IsdB, is influenced by isomerization of Pro173. These results greatly enhance current understanding of structural and dynamic aspects of the heme extraction mechanism by IsdB and provide insight into the interactions that stabilize the complex before the heme transfer event. This information will support future efforts to identify inhibitors of heme acquisition by S. aureus by interfering with IsdB:Hb complex formation.