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Context: Upper airway suctioning at birth was considered standard procedure and is still commonly practiced. Negative effects could exceed benefits of suction., Question: In infants born through clear amniotic fluid (P) does suctioning of the mouth and nose (I) vs no suctioning (C) improve outcomes (O)., Data Sources: Information specialist conducted literature search (12th September 2021, re-run 17th June 2022) using Medline, Embase, Cochrane Databases, Database of Abstracts of Reviews of Effects, and CINAHL. RCTs, non-RCTs and observational studies with a defined selection strategy were included. Unpublished studies, reviews, editorials, animal and manikin studies were excluded., Data Extraction: Two authors independently extracted data, risk of bias was assessed using the Cochrane ROB2 and ROBINS-I tools. Certainty of evidence was assed using the GRADE framework. Review Manager was used to analyse data and GRADEPro to develop summary of evidence tables. Meta-analyses were performed if ≥2 RCTs were available., Outcomes: Primary: assisted ventilation. Secondary: advanced resuscitation, oxygen supplementation, adverse effects of suctioning, unanticipated NICU admission., Results: Nine RCTs (n = 1096) and 2 observational studies (n = 418) were identified. Two RCTs (n = 280) with data concerns were excluded post-hoc. Meta-analysis of 3 RCTs, (n = 702) showed no difference in primary outcome. Two RCTs (n = 200) and 2 prospective observational studies (n = 418) found lower oxygen saturations in first 10 minutes of life with suctioning. Two RCTs (n = 200) showed suctioned newborns took longer to achieve target saturations., Limitations: Certainty of evidence was low or very low for all outcomes. Most studies selected healthy newborns limiting generalisability and insufficient data was available for planned subgroup analyses., Conclusions: Despite low certainty evidence, this review suggests no clinical benefit from suctioning clear amniotic fluid from infants following birth, with some evidence suggesting a resulting desaturation. These finding support current guideline recommendations that this practice is not used as a routine step in birth., Funding: The International Liaison Committee on Resuscitation provided access to software platforms, an information specialist and teleconferencing., Clinical Trial Registration: This systematic review was registered with the Prospective Register of Systematic Reviews (https://www.crd.york.ac.uk/prospero/) (identifier: CRD42021286258)., (© 2022 The Author(s).)

T cells depend on the phosphatase CD45 to initiate T cell receptor signaling. Although the critical role of CD45 in T cells is established, the mechanisms controlling function and localization in the membrane are not well understood. Moreover, the regulation of specific CD45 isoforms in T cell signaling remains unresolved. By using unbiased mass spectrometry, we identify the tetraspanin CD53 as a partner of CD45 and show that CD53 controls CD45 function and T cell activation. CD53-negative T cells (Cd53 -/- ) exhibit substantial proliferation defects, and Cd53 -/- mice show impaired tumor rejection and reduced IFNγ-producing T cells compared with wild-type mice. Investigation into the mechanism reveals that CD53 is required for CD45RO expression and mobility. In addition, CD53 is shown to stabilize CD45 on the membrane and is required for optimal phosphatase activity and subsequent Lck activation. Together, our findings reveal CD53 as a regulator of CD45 activity required for T cell immunity., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: In previous clinical trials, patients with active rheumatoid arthritis (RA) treated with upadacitinib (UPA) have improved patient-reported outcomes (PROs). This post hoc analysis of SELECT-CHOICE, a phase 3 clinical trial, evaluated the impact of UPA vs abatacept (ABA) with background conventional synthetic disease-modifying antirheumatic drugs (csDMARDs) on PROs in patients with RA with inadequate response or intolerance to biologic disease-modifying antirheumatic drugs (bDMARD-IR)., Methods: Patients in SELECT-CHOICE received UPA (oral 15 mg/day) or ABA (intravenous). PROs evaluated included Patient Global Assessment of Disease Activity (PtGA) by visual analog scale (VAS), patient's assessment of pain by VAS, Health Assessment Questionnaire Disability Index (HAQ-DI), morning stiffness duration and severity, 36-Item Short Form Health Survey (SF-36), Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-F), Work Productivity and Activity Impairment (WPAI), and EQ-5D 5-Level (EQ-5D-5L) index score. Least squares mean (LSM) changes from baseline to weeks 12 and 24 were based on an analysis of covariance model. Proportions of patients reporting improvements ≥ minimal clinically important differences (MCID) were compared using chi-square tests., Results: Data from 612 patients were analyzed (UPA, n=303; ABA, n=309). Mean age was 56 years and mean disease duration was 12 years. One-third received ≥2 prior bDMARDs and 72% received concomitant methotrexate at baseline. At week 12, UPA- vs ABA-treated patients had significantly greater improvements in PtGA, pain, HAQ-DI, morning stiffness severity, EQ-5D-5L, 2/4 WPAI domains, and 3/8 SF-36 domains and Physical Component Summary (PCS) scores (P<0.05); significant differences persisted at week 24 for HAQ-DI, morning stiffness severity, SF-36 PCS and bodily pain domain, and WPAI activity impairment domain. At week 12, significantly more UPA- vs ABA-treated patients reported improvements ≥MCID in HAQ-DI (74% vs 64%) and SF-36 PCS (79% vs 66%) and 4/8 domain scores (P<0.05)., Conclusions: At week 12, UPA vs ABA treatment elicited greater improvements in key domains of physical functioning, pain, and general health and earlier improvements in HAQ-DI. Overall, more UPA- vs ABA-treated patients achieved ≥MCID in most PROs at all timepoints; however, not all differences were statistically significant. These data, however, highlight the faster response to UPA treatment., Trial Registration: NCT03086343 , March 22, 2017., (© 2022. The Author(s).)

Phagocytes migrate into tissues to combat infection and maintain tissue homeostasis. As dysregulated phagocyte migration and function can lead to inflammation or susceptibility to infection, identifying molecules that control these processes is critical. Here, we show that the tetraspanin CD82 restrains the migration of neutrophils and macrophages into tissues. Cd82 -/- phagocytes exhibited excessive migration during in vivo models of peritoneal inflammation, superfusion of CXCL1, retinopathy of prematurity, and infection with the protozoan parasite L. mexicana . However, with the latter, while Cd82 -/- macrophages infiltrated infection sites at higher proportions, cutaneous L. mexicana lesions were larger and persisted, indicating a failure to control infection. Analyses of in vitro bone-marrow-derived macrophages showed CD82 deficiency altered cellular morphology, and impaired gene expression and metabolism in response to anti-inflammatory activation. Altogether, this work reveals an important role for CD82 in restraining phagocyte infiltration and mediating their differentiation in response to stimulatory cues., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)

Purpose: This study provides the first experimental application of multiscale 3-dimensional (3D) x-ray phase contrast imaging computed tomography (XPCI-CT) virtual histology for the inspection and quantitative assessment of the late-stage effects of radio-induced lesions on lungs in a small animal model., Methods and Materials: Healthy male Fischer rats were irradiated with x-ray standard broad beams and microbeam radiation therapy, a high-dose rate (14 kGy/s), FLASH spatially fractionated x-ray therapy to avoid beamlet smearing owing to cardiosynchronous movements of the organs during the irradiation. After organ dissection, ex vivo XPCI-CT was applied to all the samples and the results were quantitatively analyzed and correlated to histologic data., Results: XPCI-CT enables the 3D visualization of lung tissues with unprecedented contrast and sensitivity, allowing alveoli, vessel, and bronchi hierarchical visualization. XPCI-CT discriminates in 3D radio-induced lesions such as fibrotic scars and Ca/Fe deposits and allows full-organ accurate quantification of the fibrotic tissue within the irradiated organs. The radiation-induced fibrotic tissue content is less than 10% of the analyzed volume for all microbeam radiation therapy-treated organs and reaches 34% in the case of irradiations with 50 Gy using a broad beam., Conclusions: XPCI-CT is an effective imaging technique able to provide detailed 3D information for the assessment of lung pathology and treatment efficacy in a small animal model., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Stimulator of Interferon Genes (STING) is a cytosolic sensor of cyclic dinucleotides (CDNs). The activation of dendritic cells (DC) via the STING pathway, and their subsequent production of type I interferon (IFN) is considered central to eradicating tumours in mouse models. However, this contribution of STING in preclinical murine studies has not translated into positive outcomes of STING agonists in phase I & II clinical trials. We therefore questioned whether a difference in human DC responses could be critical to the lack of STING agonist efficacy in human settings. This study sought to directly compare mouse and human plasmacytoid DCs and conventional DC subset responses upon STING activation. We found all mouse and human DC subsets were potently activated by STING stimulation. As expected, Type I IFNs were produced by both mouse and human plasmacytoid DCs. However, mouse and human plasmacytoid and conventional DCs all produced type III IFNs (i.e., IFN-λs) in response to STING activation. Of particular interest, all human DCs produced large amounts of IFN-λ1, not expressed in the mouse genome. Furthermore, we also found differential cell death responses upon STING activation, observing rapid ablation of mouse, but not human, plasmacytoid DCs. STING-induced cell death in murine plasmacytoid DCs occurred in a cell-intrinsic manner and involved intrinsic apoptosis. These data highlight discordance between STING IFN and cell death responses in mouse and human DCs and caution against extrapolating STING-mediated events in mouse models to equivalent human outcomes., Competing Interests: HH is an employee of Bavarian Nordic GmbH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Pang, Daraj, Balka, De Nardo, Macri, Hochrein, Masterman, Tan, Shoppee, Magill, Jahan, Bafit, Zhan, Kile, Lawlor, Radford, Wright and O’Keeffe.)

Tetraspanins are proteins that organize cell membranes via interactions with partner proteins mediated by their large ectodomain. In this issue of Structure, Lipper et al., 2022 have elucidated the structure of the first C8 tetraspanin and expand functional insight into how C8 tetraspanins regulate substrate specificity for the transmembrane protease ADAM10., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Using Brofenbrenner's socio-ecological model as a conceptual framework, the objective of this study was to determine playground users (primary school staff and pupils) perceptions of the barriers and facilitators to a physically active school playground at an intra-personal (individual), inter-personal (social), environmental and policy level. Results from a series of qualitative interactions with children (n = 65) from years five and six (9 to 11 years old), and structured interviews with adult teachers (n = 11) revealed key differences in the child and adult perceptions of the playground and the purpose of break-times. A number of inter-related environmental boundaries and school policies were identified as restrictive to children's explorations and activity levels during 'free play' periods, which centred on resource availability, accessibility and health and safety. Further, traditional playground hierarchies act to promote and prevent physical activity engagement for different groups (e.g. gender and age). Finally, differences between the adult and child perception of the primary school playground were observed. Playground physical activity, during break-times appears to be affected by a number of variables at each level of the socio-ecological model. This study provides an opportunity for primary schools to reflect on primary school playground strategies and practices that are implemented at each level of the socio-ecological model to encourage a more effective use of the playground during school break-times., Competing Interests: The authors have declared that no competing interests exist.

Conventional radiotherapy is a widely used non-invasive form of treatment for many types of cancer. However, due to a low threshold in the lung for radiation-induced normal tissue damage, it is of less utility in treating lung cancer. For this reason, surgery is the preferred treatment for lung cancer, which has the detriment of being highly invasive. Non-conventional ultra-high dose rate (FLASH) radiotherapy is currently of great interest in the radiotherapy community due to demonstrations of reduced normal tissue toxicity in lung and other anatomy. This study investigates the effects of FLASH microbeam radiotherapy, which in addition to ultra-high dose rate incorporates a spatial segmentation of the radiation field, on the normal lung tissue of rats. With a focus on fibrotic damage, this work demonstrates that FLASH microbeam radiotherapy provides an order of magnitude increase in normal tissue radio-resistance compared to FLASH radiotherapy. This result suggests FLASH microbeam radiotherapy holds promise for much improved non-invasive control of lung cancer., Competing Interests: Two U. S. patents issued to author MDW as Inventor and to VMS as Assignee are related to the work., (Copyright © 2021, Wright et al.)

The leukocyte-restricted tetraspanin CD53 has been shown to promote lymphocyte homing to lymph nodes (LNs) and myeloid cell recruitment to acutely inflamed peripheral organs, and accelerate the onset of immune-mediated disease. However, its contribution in the setting of chronic systemic autoimmunity has not been investigated. We made use of the Lyn -/- autoimmune model, generating Cd53 -/- Lyn -/- mice, and compared trafficking of immune cells into secondary lymphoid organs and systemic autoimmune disease development with mice lacking either gene alone. Consistent with previous observations, absence of CD53 led to reduced LN cellularity via reductions in both B and T cells, a phenotype also observed in Cd53 -/- Lyn -/- mice. In some settings, Cd53 -/- Lyn -/- lymphocytes showed greater loss of surface L-selectin and CD69 upregulation above that imparted by Lyn deficiency alone, indicating that absence of these two proteins can mediate additive effects in the immune system. Conversely, prototypical effects of Lyn deficiency including splenomegaly, plasma cell expansion, elevated serum immunoglobulin M and anti-nuclear antibodies were unaffected by CD53 deficiency. Furthermore, while Lyn -/- mice developed glomerular injury and showed elevated glomerular neutrophil retention above than that in wild-type mice, absence of CD53 in Lyn -/- mice did not alter these responses. Together, these findings demonstrate that while tetraspanin CD53 promotes lymphocyte trafficking into LNs independent of Lyn, it does not make an important contribution to development of autoimmunity, plasma cell dysfunction or glomerular injury in the Lyn -/- model of systemic autoimmunity., (© 2021 Australian and New Zealand Society for Immunology Inc.)

Abstract: People living with HIV (PLWH) are at increased risk for cardiovascular disease. Physical activity, exercise, and controlled diet can mitigate this risk, yet these behaviors are understudied in sub-Saharan Africa. Our objective was to describe and compare the meaning, value, and patterns of physical activity, exercise, and diet among PLWH and older adults without HIV in Uganda. This mixed methods, observational study included 30 adult PLWH and 29 adults without HIV who (a) wore an accelerometer to measure physical activity; (b) had weight, height, and waist and hip circumference measured; (c) completed physical fitness measures; and (d) used digital cameras to record photographs and videos of their typical diet and physical activities. Participants were approximately 58 years old and 68% female. Approximately 20% of PLWH and 40% of adults without HIV met physical activity guidelines (p > .05). Qualitative themes included engaging in a variety of exercise, structural barriers to exercising, and typical meals. Older adults in Uganda have low levels of physical activity and homogenous diets, increasing their risk for cardiovascular disease., (Copyright © 2020 Association of Nurses in AIDS Care.)

Background: Invasive lobular carcinoma (ILC) is thought be a unique entity with higher rates of multifocal/multicentric and bilateral disease. This study aimed to evaluate the true extent of the disease, risk of bilaterality, lymph node involvement, and impact of preoperative imaging to help guide surgical decision making., Methods: A retrospective analysis identified patients treated for ILC between 2004 and 2017. Clinical staging and pathologic results were compared. Follow-up details including local recurrence, contralateral breast cancer (CBC), and survival outcomes were evaluated., Results: The study identified 692 patients with ILC, including 43 patients (6%) with a diagnosis of CBC and 232 patients (33%) with a diagnosis of multifocal/multicentric disease at presentation. Preoperative magnetic resonance imaging (MRI) led to an identification of additional disease in 20% of the patients. Preoperative MRI resulted in a more accurate prediction of tumor size staging but did not improve the discordance between clinical and pathologic nodal staging. Overall, the rate of imaging occult lymph node disease was 24%. At the 6-year follow-up evaluation, a local recurrence had developed in 2.3%, a CBC in 2.3, and a distant metastasis in 9.4% of the patients. The overall survival rate was 96% at 3 years and 91% at 5 years., Conclusions: Invasive lobular carcinoma is a distinct subset of cancer that poses a diagnostic staging challenge. The results of this study favor MRI for accurate tumor staging and for improving detection of multicentricity and bilaterality. However, clinicians should be aware of the higher likelihood of occult lymph node involvement with ILC and subsequent early metastasis., (© 2021. Society of Surgical Oncology.)

Background: Home blood pressure monitoring is increasingly used for pregnant individuals; however, there are no guidelines on such monitoring in this population. We assessed current practices in the prescription and use of home blood pressure monitoring in pregnancy., Methods: We conducted a systematic review and meta-analysis of observational studies and randomized controlled trials (RCTs). We conducted a structured search through the MEDLINE (from 1946), Embase (from 1974) and CENTRAL (from 2018) databases up to Oct. 19, 2020. We included trials comparing office and home blood pressure monitoring in pregnant people. Outcomes included patient education, home blood pressure device, monitoring schedule, adherence, diagnostic thresholds for home blood pressure, and comparison between home and office measurements of blood pressure., Results: We included in our review 21 articles on 19 individual studies (1 RCT, 18 observational) that assessed home and office blood pressure in pregnant individuals ( n = 2843). We observed variation in practice patterns in terms of how home monitoring was prescribed. Eight (42%) of the studies used validated home blood pressure devices. Across all studies, measurements were taken 3 to 36 times per week. Third-trimester home blood pressure corresponding to office blood pressure of 140/90 mm Hg after application of a conversion factor ranged from 118 to 143 mm Hg (systolic) and from 76 to 92 mm Hg (diastolic), depending on the patient population and methodology. Systolic and diastolic blood pressure values measured at home were lower than office values by 4 (95% confidence interval [CI] -6 to -3) mm Hg and 3 (95% CI -4 to -2) mm Hg, respectively., Interpretation: Many issues related to home blood pressure monitoring in pregnancy are currently unresolved, including technique, monitoring schedule and target values. Future studies should prioritize the use of validated home measuring devices and standardized measurement schedules and should establish treatment targets., Prospero Registration: CRD42020147352., Competing Interests: Competing interests: Raj Padwal is the CEO of mmHg Inc., provider of cloud-based digital health solutions. No other competing interests were declared., (© 2021 CMA Joule Inc. or its licensors.)

Introduction : Cyclin-dependent kinase (CDK) 4/6 inhibitors have altered the standard-of-care treatment for patients with ER-positive, HER2-negative metastatic breast cancer. One such inhibitor, abemaciclib, a reversible ATP-competitive CDK4/6 inhibitor developed by Eli Lilly and Company, was approved by the FDA for ER-positive, HER2-negative metastatic breast cancer. Areas covered : Preclinical studies revealed abemaciclib's distinct structure, efficacy as monotherapy, and ability to penetrate the Central Nervous System. In this review, the authors have examined the literature regarding the development of CDK 4/6 inhibitors before providing a focused review on the preclinical discovery and development of abemaciclib. The authors then conclude their manuscript by providing their expert opinion and future perspectives. Expert opinion : Understanding the genesis and evolution from concept to approval and beyond will allow one to analyze the impact of abemaciclib. With its unique characteristics, abemaciclib has provided a meaningful addition to the therapeutic arsenal for metastatic breast cancer. There is, however, a need for predictive biomarkers to identify patients who may not benefit from or may develop resistance to CDK4/6 inhibition.

Opioid use disorder has long been associated with psychiatric symptoms, including dissociative experiences. Medications used to treat opioid use disorder can potentially impact dissociative symptoms, but the existing literature has not explored this. We examined the relationship between dissociative symptoms and opioid use disorder using the Dissociative Experiences Scale (DES). We studied subjects who were taking prescribed methadone, buprenorphine, or naltrexone for opioid use disorder. We gave the DES, the Patient Health Questionairre-9 (PHQ-9), and the PTSD Checklist for DSM-5 (PCL-5) with Criterion A to subjects in three substance use treatment facilities in Ohio. We conducted Analysis of Variance (ANOVA) and Spearman's Rank Correlations to examine associations between the variables and outcomes. We developed three separate multiple linear regression models. We included 116 participants in our exploratory and naturalistic study. The majority of participants were female (51.7%), white (89.5%), ≤ 40 years of age (64.7%), and taking buprenorphine (55%). The average DES score was 16.1 (standard deviation = 14.9) and we considered 80.9% to have low dissociation (score < 30). Approximately 55% (n = 64) of participants were taking prescribed buprenorphine. Approximately 27% (n = 32) were taking prescribed methadone and approximately 18% (n = 21) were taking prescribed naltrexone (oral or extended release). There was a significant association between opioid medication type and log dissociative symptoms (p = .01). Participants taking prescribed buprenorphine had higher mean log dissociation symptom scores (2.7) compared to those taking prescribed methadone (2.2) and prescribed naltrexone (2.1). Log dissociation symptom scores were significantly associated with last use of any opiates (rs = -0.21; p = .02) and time on medication (rs = -0.228; p = .01). Compared to those taking buprenorphine, those taking both methadone (β = -0.26; p = .01) and naltrexone (β = -0.27; p = .006) had significantly lower dissociation scores, controlling for the other variables in the model. Dissociation scores were positively correlated with depression scores (r = 0.45; p < .0001) and with PCL-5 scores (r = 0.51; p < .0001). Our study highlights the importance of diagnosing and monitoring dissociative symptoms in individuals who are taking prescribed medications for opioid use disorder, especially since dissociative symptoms can interfere with substance use treatment., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Immunity plays a key role in epithelial ovarian cancer (EOC) progression with a well-documented correlation between patient survival and high intratumoral CD8+ to T regulatory cell (Treg) ratios. We previously identified dysregulated DPP4 activity in EOCs as a potentially immune-disruptive influence contributing to a reduction in CXCR3-mediated T-cell infiltration in solid tumours. We therefore hypothesized that inhibition of DPP4 activity by sitagliptin, an FDA-approved inhibitor, would improve T-cell infiltration and function in a syngeneic ID8 mouse model of EOC. Daily oral sitagliptin at 50 mg/kg was provided to mice with established primary EOCs. Sitagliptin treatment decreased metastatic tumour burden and significantly increased overall survival and was associated with significant changes to the immune landscape. Sitagliptin increased overall CXCR3-mediated CD8+ T-cell trafficking to the tumour and enhanced the activation and proliferation of CD8+ T-cells in tumour tissue and the peritoneal cavity. Substantial reductions in suppressive cytokines, including CCL2, CCL17, CCL22 and IL-10, were also noted and were associated with reduced CD4+ CD25+ Foxp3+ Treg recruitment in the tumour. Combination therapy with paclitaxel, however, typical of standard-of-care for patients in palliative care, abolished CXCR3-specific T-cell recruitment stimulated by sitagliptin. Our data suggest that sitagliptin may be suitable as an adjunct therapy for patients between chemotherapy cycles as a novel approach to enhance immunity, optimise T-cell-mediated function and improve overall survival.

Objectives: Our objectives were to examine the impact of methamphetamine use on opioid use disorder (OUD) treatment retention in patients prescribed either buprenorphine/buprenorphine-naloxone (BUP-NX) or naltrexone/extended-release naltrexone (XR-NTX), while also exploring the role of other risk factors that may modify the impact of methamphetamine use., Methods: We conducted an exploratory retrospective study examining OUD treatment retention in 127 patients in Ohio (USA). Patients were prescribed either BUP-NX or naltrexone/XR-NTX. Cox proportional hazard regression was used to compare time to dropout of treatment between patients positive and negative on screening for methamphetamines at intake, estimate the association between other risk factors and time to dropout, and test interactions between risk factors and methamphetamine status., Results: Among patients prescribed naltrexone/XR-NTX, those positive for methamphetamines had almost three times the risk of treatment dropout (AHR = 2.89, 95% CI =1.11, 7.07), significantly greater (interaction p = .039) than the methamphetamine effect among those prescribed BUP-NX (AHR = 0.94, 95% CI = 0.51, 1.65). Early in treatment, being prescribed BUP-NX was strongly associated with a greater risk of treatment dropout (at baseline: AHR = 2.90, 95% CI = 1.33, 7.15), regardless of baseline methamphetamine use status. However, this effect decreased with time and shifted to greater risk of dropout among those prescribed naltrexone/XR-NTX (non-proportional hazard; interaction with time AHR = 0.66, 95% CI = 0.49, 0.86), with the shift occurring sooner among those positive for methamphetamine at baseline., Conclusions: Additional support should be provided to patients who use methamphetamines prior to starting OUD treatment.

The dendritic cell receptor Clec9A facilitates processing of dead cell-derived antigens for cross-presentation and the induction of effective CD8 + T cell immune responses. Here, we show that this process is regulated by E3 ubiquitin ligase RNF41 and define a new ubiquitin-mediated mechanism for regulation of Clec9A, reflecting the unique properties of Clec9A as a receptor specialized for delivery of antigens for cross-presentation. We reveal RNF41 is a negative regulator of Clec9A and the cross-presentation of dead cell-derived antigens by mouse dendritic cells. Intriguingly, RNF41 regulates the downstream fate of Clec9A by directly binding and ubiquitinating the extracellular domains of Clec9A. At steady-state, RNF41 ubiquitination of Clec9A facilitates interactions with ER-associated proteins and degradation machinery to control Clec9A levels. However, Clec9A interactions are altered following dead cell uptake to favor antigen presentation. These findings provide important insights into antigen cross-presentation and have implications for development of approaches to modulate immune responses., Competing Interests: KT, PT, HP, RS, NM, RL, AP, EG, CH, AF, JD, PC, LW, JM, GR, KR, IC, MO, JV, MW, MB, WH, KS, AP, NN, JZ, ML No competing interests declared, (© 2020, Tullett et al.)

Nanoparticles have been incorporated into a range of consumer spray products, providing the potential for inadvertent inhalation by users and bystanders. The levels and characteristics of nanoparticle inhalation exposures arising from the use of such products are important inputs to risk assessments and informing dose regimes for in vitro and in vivo studies investigating hazard potentials. To date, only a small number of studies have been undertaken to explore both the aerosols generated from such products and the metal nanoparticles within them. The objective of the current study was to add to the limited data in this field by investigating a range of nano-containing spray products available within the UK. Six products were selected and the nanoparticles characterised using a combination of techniques, including: inductively coupled plasma mass spectrometry (ICP-MS), dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), transmission electron microscopy energy-dispersive X-ray spectroscopy (TEM-EDX) and single particle ICP-MS (spICP-MS). The aerosol produced by these products, when sprayed within a glovebox, was characterised by scanning mobility particle sizer (SMPS) and an aerodynamic particle sizer (APS). A cascade impactor with thirteen stages (NanoMOUDI) was used with one product to generate information on the size specific nanoparticle elemental distribution within the aerosol. The results demonstrated the presence of solid nanoparticles (silver, gold or silica) in each of the products at low concentrations (<13 ppm). TEM and (sp)ICP-MS provided reliable information on nanoparticle size, shape, number and mass, while the light scattering methods were less effective due to the complex matrices of the products and their lack of chemical specificity. The aerosols varied significantly across products, with particle and mass concentrations spanning 5 orders of magnitude (10 - 10 6  cm -3 and 0.3-7600 μg m -3 , respectively). The NanoMOUDI results clearly indicated non-uniform distribution of silver within different aerosol particle size ranges., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2020 The Authors.)

Purpose: To understand the validity of differential ratings of perceived exertion (dRPE) as a measure of girls' training and match internal loads., Methods: Using the centiMax scale (CR100), session dRPE for breathlessness (sRPE-B) and leg muscle exertion (sRPE-L) were collected across a season of training (soccer, resistance, and fitness) and matches from 33 players (15 [1] y). Differences and associations between dRPE were examined using mixed and general linear models. The authors' minimal practical important difference was 8 arbitrary units (AU)., Results: Mean (AU [SD] ∼16) sRPE-B and sRPE-L were 66 and 61 for matches, 51 and 49 for soccer, 86 and 67 for fitness, and 45 and 58 for resistance, respectively. Session RPE-B was rated most likely harder than sRPE-L for fitness (19 AU; 90% confidence limits: ±7) and most likely easier for resistance (-13; ±2). Match (5; ±4) and soccer (-3; ±2) differences were likely to most likely trivial. The within-player relationships between sRPE-B and sRPE-L were very likely moderate for matches (r = .44; 90% confidence limits: ±.12) and resistance training (.38; ±.06), likely large for fitness training (.51; ±.22), and most likely large for soccer training (.56; ±.03). Shared variance ranged from 14% to 35%., Conclusions: Practically meaningful differences between dRPE following physical training sessions coupled with low shared variance in all training types and matches suggest that sRPE-B and sRPE-L represent unique sensory inputs in girls' soccer players. The data provide evidence for the face and construct validity of dRPE as a measure of internal load in this population.

The importance of tetraspanin proteins in regulating migration has been demonstrated in many diverse cellular systems. However, the function of the leukocyte-restricted tetraspanin CD53 remains obscure. We therefore hypothesized that CD53 plays a role in regulating leukocyte recruitment and tested this hypothesis by examining responses of CD53-deficient mice to a range of inflammatory stimuli. Deletion of CD53 significantly reduced neutrophil recruitment to the acutely inflamed peritoneal cavity. Intravital microscopy revealed that in response to several inflammatory and chemotactic stimuli, absence of CD53 had only minor effects on leukocyte rolling and adhesion in postcapillary venules. In contrast, Cd53 -/- mice showed a defect in leukocyte transmigration induced by TNF, CXCL1 and CCL2, and a reduced capacity for leukocyte retention on the endothelial surface under shear flow. Comparison of adhesion molecule expression in wild-type and Cd53 -/- neutrophils revealed no alteration in expression of β 2 integrins, whereas L-selectin was almost completely absent from Cd53 -/- neutrophils. In addition, Cd53 -/- neutrophils showed defects in activation-induced cytoskeletal remodeling and translocation to the cell periphery, responses necessary for efficient transendothelial migration, as well as increased α 3 integrin expression. These alterations were associated with effects on inflammation, so that in Cd53 -/- mice, the onset of neutrophil-dependent serum-induced arthritis was delayed. Together, these findings demonstrate a role for tetraspanin CD53 in promotion of neutrophil transendothelial migration and inflammation, associated with CD53-mediated regulation of L-selectin expression, attachment to the endothelial surface, integrin expression and trafficking, and cytoskeletal function., (Copyright © 2020 by The American Association of Immunologists, Inc.)

Carbon nanotube (CNT) in vivo inhalation studies are increasingly providing estimates of the quantity of material deposited in the lung, generally estimated using standard formulae and pulmonary deposition models. These models have typically been developed and validated using data from studies using sphere-like particles. Given the importance of particle morphology to pulmonary deposition, the appropriateness of such an approach was explored to identify any potential limitations. Aerosolized CNT particles typically form 'fiber-like' and/or 'broadly spherical' agglomerates. A review of currently used deposition models indicates that none have been directly validated against results for CNT, however, models for spherical particles have been extensively validated against a wide range of particle sizes and materials and are thus expected to provide reasonable estimates for most 'broadly spherical' CNT particles, although experimental confirmation of this would be of benefit, especially given their low density. The validation of fiber deposition models is significantly less extensive and, in general, focused on larger particles, e.g. asbestos. This raises concerns about the accuracy of deposition estimates for 'fiber-like' CNT particles and recommendations are made for future research to address this. An appreciation of the uncertainties on CNT deposition estimates is important for their interpretation and thus it is recommended that model sensitivity and uncertainty assessments be undertaken. Issues surrounding the measurement and derivation of model input data are also addressed, including instrument responses and particle density assessment options. Recommendations are also made for aerosol characterization to 'future-proof' CNT inhalation studies regarding advances in deposition modeling and toxicological understanding.

Tetraspanins regulate key processes in immune cells; however, the function of the leukocyte-restricted tetraspanin CD53 is unknown. Here we show that CD53 is essential for lymphocyte recirculation. Lymph nodes of Cd53 -/- mice were smaller than those of wild-type mice due to a marked reduction in B cells and a 50% decrease in T cells. This reduced cellularity reflected an inability of Cd53 -/- B and T cells to efficiently home to lymph nodes, due to the near absence of L-selectin from Cd53 -/- B cells and reduced stability of L-selectin on Cd53 -/- T cells. Further analyses, including on human lymphocytes, showed that CD53 stabilizes L-selectin surface expression and may restrain L-selectin shedding via both ADAM17-dependent and ADAM17-independent mechanisms. The disruption in lymphocyte recirculation in Cd53 -/- mice led to impaired immune responses dependent on antigen delivery to lymph nodes. Together these findings demonstrate an essential role for CD53 in lymphocyte trafficking and immunity., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)

Urban aerosol is a growing concern for people living within cities; aerosol have been implicated in many ill health conditions, including that of the lung and of the heart. Atmospheric potential gradient is a consequence of charge carried to the ionosphere through thunderstorms, and its value depends on highly electrically mobile ion concentrations, hence local conductivity of the air. Ions attach to aerosol in the atmosphere, reducing their mobility and therefore increasing the potential gradient, and so potential gradient measurements have been suggested as a proxy for aerosol measurements. Particle number count, size distribution and potential gradient were measured for two campaigns in Manchester, U.K., and one campaign in Bristol, U.K. Using a factor based on size distribution to account for preferential attachment at larger sizes provided the best relationship with potential gradient, but particle count alone showed a weaker, but similar relationship. The increase in particle count caused by annual bonfire and fireworks celebrations (November) was evidenced in both potential gradient and particle numbers. Daily regression or correlation did not show a consistent relationship. In the larger Bristol data set, increasing humidity led to a reduction of potential gradient, while increasing particle number led to an increase., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

We aimed to evaluate the internal consistency and factor structure of the Functional Movement Screen (FMS TM ) in youth athletes and quantify differences between individual task score at different stages of maturation. FMS TM and anthropometric variables were measured in 144 youth athletes (96 female, 48 male). Biological maturation was categorised as before- (<-0.5 years), at- (-0.49-0.50 years) and after- peak height velocity [PHV] (>0.51 years). Internal consistency was poor (Cronbach's alpha; 0.53, ±90% confidence limit 0.10; ordinal alpha 0.6, ±0.09). Principle component analysis extracted two components, representing 47% of the total variance. Tasks loading highest on component 1 required stability, while those loading highest on component 2 favoured mobility. "Likely" decrements in component 1 tasks were observed before-PHV. In-line lunge (effect size ±90% confidence limit; -0.47, ±0.49), hurdle step (-0.38, ±0.49), and trunk stability push-up (-0.51, ±0.45), were lower compared with athletes at-PHV and rotatory stability (-0.44, ±0.37) was lower than those after-PHV. Boys' scored "most likely", higher (0.73, ±0.28) in trunk stability push-up, and girls "likely" higher in shoulder mobility (0.46, ±0.29). In our population, the FMS TM is not uni-dimensional, thus total score should be avoided. Clear maturation affects were observed in stability tasks.

Wright, MD and Atkinson, G. Changes in sprint-related outcomes during a period of systematic training in a girls' soccer academy. J Strength Cond Res 33(3): 793-800, 2019-The longitudinal tracking of performance indicators is lacking during the training of young female soccer players. Therefore, changes in 5-m acceleration, 20-m speed, change-of-direction (COD) speed, and repeated-sprint ability (RSA) were quantified during a 3-year period in an English Football Association Center of Excellence. Fourteen players (mean age = 12.1 years, SD = ±0.9) were recruited, and their best performance scores from preseason and in-season testing were averaged. Players were typically exposed to soccer (2 × 90 minutes per week) and strength and conditioning training (1 × 70 minutes per week) and played 20 soccer matches (50-80 minutes) during 35-week seasons. Mean (±90% confidence limits (CL)) overall improvements over the 3 years were 5.9% (1.3) (most likely large) for speed, 4.0% (1.0) (most likely large) for RSA, 8.8% (1.1) for acceleration, and 8.3% (1.4) for COD speed (both most likely very large). Improvements between years one and 2 ranged from most likely moderate to very large. Further small improvements in COD speed and 20-m speed (both likely) were observed between years 2 and 3. Individual differences in response were apparent only for COD speed, which were moderate and small between years 2 and 3. Most likely very large to near-perfect within-player correlations were observed between maturation and sprint measures. These data from a single-arm longitudinal study indicate that systematic exposure to training, which includes one dedicated strength and conditioning session each week, is associated with improvements in sprint-related physical qualities in girls.

Immune cell recruitment and migration is central to the normal functioning of the immune system in health and disease. Numerous adhesion molecules on immune cells and the parenchymal cells they interact with are well recognized for their roles in facilitating the movements of immune cells throughout the body. A growing body of evidence now indicates that tetraspanins, proteins known for their capacity to organize partner molecules within the cell membrane, also have significant impacts on the ability of immune cells to migrate around the body. In this review, we examine the tetraspanins expressed by immune cells and endothelial cells that influence leukocyte recruitment and motility and describe their impacts on the function of adhesion molecules and other partner molecules that modulate the movements of leukocytes. In particular, we examine the functional roles of CD9, CD37, CD63, CD81, CD82, and CD151. This reveals the diversity of the functions of the tetraspanin family in this setting, both in the nature of adhesive and migratory interactions that they regulate, and the positive or inhibitory effects mediated by the individual tetraspanin proteins.

Objective: To characterize the effect of anti-tumor necrosis factor (TNF) therapy compared to conventional synthetic disease-modifying antirheumatic drugs (csDMARD) in children with enthesitis-related arthritis (ERA) over the first year after diagnosis., Methods: We conducted a multicenter retrospective comparative effectiveness study of children diagnosed with ERA. We estimated the effect of anti-TNF therapy on clinical variables (active joint count, tender entheses count) and patient-reported pain and global assessment of disease activity over the first year after diagnosis using state-of-the-art comparative effectiveness analytic methods., Results: During the study period, 217 patients newly diagnosed with ERA had a total of 965 clinic visits the first year after disease diagnosis. Children [median age 11.6 yrs, interquartile range 10-14] were treated with anti-TNF monotherapy (n = 33, 15.2%), csDMARD monotherapy (n = 73, 33.6%), or both (n = 52, 23.9%) in the first year after disease diagnosis. There was a statistically significant improvement in the primary outcome, active joint count, over time in children who received an anti-TNF drug versus those who did not (p = 0.03). Additionally, use of anti-TNF therapy versus no anti-TNF therapy was associated with less patient-reported pain (p < 0.01) and improved disease activity over time as assessed by the clinical Juvenile Arthritis Disease Activity Score (p < 0.01). The magnitude of estimated effect on clinical outcomes was uniformly greater, with the exception of tender entheses count, in children treated with an anti-TNF drug versus a csDMARD., Conclusion: During the first year after diagnosis, anti-TNF exposure was associated with benefits for several clinically meaningful outcomes in children with enthesitis-related arthritis.

Tumor-suppressing subchromosomal transferable fragment cDNA 6 (TSSC6) expression is restricted to hematopoietic organs and tissues where it plays a role in hematopoietic-cell function. The ADP purinergic receptor P2Y 12 is mainly expressed by platelets with important clinical significance as a target for several clinically approved antithrombotic agents. We have previously shown a physical association between P2Y 12 and TSSC6 in platelets. Hence our aim was to investigate whether this physical association is translated to functional effects. To investigate this possibility, we used wild-type or TSSC6 knockout (KO) mice treated with either PBS or 50mg/kg clopidogrel. TSSC6 KO mice treated with clopidogrel exhibited synergy in delayed kinetics of clot retraction, reduced collagen-mediated platelet aggregation, and platelet spreading on fibrinogen. Platelets derived from TSSC6 mice with P2Y 12 blockade form smaller thrombi when perfused over a collagen matrix under arterial flow. Clopidogrel treated TSSC6 KO arterioles showed smaller and less stable thrombi with increased tendency to embolise in vivo. These studies demonstrate a complementary role between TSSC6 and P2Y 12 receptor in platelets in regulating 'outside in' integrin α IIb β 3 signalling thrombus growth and stability., (Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.)

Objective: To describe long-term outcomes of children with early-infantile Krabbe disease who underwent hematopoietic stem cell transplantation (HSCT) in the first 7 weeks of life., Methods: In this prospective longitudinal study, evaluations performed at baseline and follow-up included brain imaging, neurodiagnostic tests, and neurobehavioral evaluations., Results: Of the 18 patients in this study (11 girls, 7 boys; mean follow-up 9.5 years, range 4-15), 5 died (3 of peritransplant complications, 1 of a surgical complication unrelated to Krabbe disease, 1 of disease progression). One of the surviving patients has normal cognitive function and 10 continue to develop cognitive skills at a slightly slower rate than normal. All surviving patients continue to gain receptive language skills, with 7 falling within the normal range. Ten patients receive speech therapy, and 2 of these patients require augmentative communication devices. Gross motor development varies widely, but 3 patients can walk independently, and 7 walk with assistive devices. Spasticity ranges from mild to severe, and 12 patients wear orthotics. Fine motor skills are generally preserved. Brain myelination and atrophy stabilized in 8 patients, improved in 4 patients, and worsened in 1 patient. Nerve conduction velocities initially improved but continue to be abnormal in most patients., Conclusions: The surviving patients function at a much higher level than untreated children or symptomatic children who underwent HSCT. These results show that early HSCT changes the natural history of this disease by improving both lifespan and functional abilities., Classification of Evidence: This study provides Class IV evidence that for children with early-infantile Krabbe disease, early HSCT improves lifespan and functional abilities., (© 2017 American Academy of Neurology.)

Activation of B cells by the binding of antigens to the B cell receptor (BCR) requires the protein kinase C (PKC) family member PKCβ. Because PKCs must translocate to the plasma membrane to become activated, we investigated the mechanisms regulating their spatial distribution in mouse and human B cells. Through live-cell imaging, we showed that BCR-stimulated production of the second messenger diacylglycerol (DAG) resulted in the translocation of PKCβ from the cytosol to plasma membrane regions containing the tetraspanin protein CD53. CD53 was specifically enriched at sites of BCR signaling, suggesting that BCR-dependent PKC signaling was initiated at these tetraspanin microdomains. Fluorescence lifetime imaging microscopy studies confirmed the molecular recruitment of PKC to CD53-containing microdomains, which required the amino terminus of CD53. Furthermore, we showed that Cd53 -deficient B cells were defective in the phosphorylation of PKC substrates. Consistent with this finding, PKC recruitment to the plasma membrane was impaired in both mouse and human CD53 -deficient B cells compared to that in their wild-type counterparts. These data suggest that CD53 promotes BCR-dependent PKC signaling by recruiting PKC to the plasma membrane so that it can phosphorylate its substrates and that tetraspanin-containing microdomains can act as signaling hotspots in the plasma membrane., (Copyright © 2017, American Association for the Advancement of Science.)

Macrophages are heterogeneous innate immune cells which are important in both the maintenance of tissue homeostasis and its disruption, by promoting tissue inflammation and fibrosis. The renin-angiotensin system is central to the pathophysiology of a large suite of diseases, which are driven in part by large amounts of tissue inflammation and fibrosis. Here, we review recent advances in understanding macrophage heterogeneity in origin and function, and how these may lead to new insights into the pathogenesis of these chronic diseases.

People spend the majority of their time indoors and therefore the quality of indoor air is worthy of investigation; indoor air quality is affected by indoor sources of pollutants and from pollutants entering buildings from outdoors. In this study, unique perfluorocarbon tracers were released in five experiments at a 100 m and ~2 km distance from a large university building in Manchester, UK and tracer was also released inside the building to measure the amount of outdoor material penetrating into buildings and the flow of material within the building itself. Air samples of the tracer were taken in several rooms within the building, and a CO₂ tracer was used within the building to estimate air-exchange rates. Air-exchange rates were found to vary between 0.57 and 10.90 per hour. Indoor perfluorocarbon tracer concentrations were paired to outdoor tracer concentrations, and in-out ratios were found to vary between 0.01 and 3.6. The largest room with the lowest air-exchange rate exhibited elevated tracer concentrations for over 60 min after the release had finished, but generally had the lowest concentrations, the room with the highest ventilation rates had the highest concentration over 30 min, but the peak decayed more rapidly. Tracer concentrations indoors compared to outdoors imply that pollutants remain within buildings after they have cleared outside, which must be considered when evaluating human exposure to outdoor pollutants.

The use of big data to transform care delivery is rapidly becoming a reality. To deliver on the promise of value-based care, providers must know the key drivers of wellness at the patient and community levels, as well as understand resource constraints and opportunities to improve efficiency in the health-care system itself. Data are the linchpin. By gathering the right data and finding innovative ways to glean knowledge, we can improve clinical care, advance the health of our communities, improve the lives of our patients, and operate more efficiently. At Carolinas HealthCare System-one of the nation's largest health-care systems, with nearly 12 million patient encounters annually at more than 900 care locations-we have made substantial investments to establish a centralized data and analytics infrastructure that is transforming the way we deliver care across the continuum. Although the impetus and vision for our program have evolved over the past decade, our efforts coalesced into a strategic, centralized initiative with the launch of the Dickson Advanced Analytics (DA) group in 2012. DA has yielded significant gains in our ability to use data, not only for reporting purposes and understanding our business but also for predicting outcomes and informing action.While these efforts have been successful, the path has not been easy. Effectively harnessing big data requires navigating myriad technological, cultural, operational, and other hurdles. Building a program that is feasible, effective, and sustainable takes concerted effort and a rigorous process of continuous self-evaluation and strategic adaptation.

Little is known about the responses of girl athletes to training interventions throughout maturation. This study evaluated group and individual responses to an 8-week, mixed-methods, high-intensity interval training (HIIT) programme in girl football players. Thirty-seven players (age 13.4 ± 1.5 years) were tested for 20-m speed, repeated-sprint ability, change-of-direction speed and level 1 yo-yo intermittent recovery (YYIR). Players were subcategorised into before-, at- and after-PHV (peak height velocity) based on maturity offset. Very likely moderate (25%; ±90% confidence limits = 9.2) improvements occurred in YYIR, but data were unclear in players before-PHV with moderate individual differences in response. Decrements in repeated-sprint ability were most likely very large (6.5%; ±3.2) before-PHV, and likely moderate (1.7%; ±1.0) at-PHV. Data were unclear after-PHV. A very likely moderate (2.7%; ±1.0) decrement occurred in change-of-direction speed at-PHV while there was a very likely increase (-2.4%; ±1.3) in after-PHV players. Possibly small (-1.1%; ±1.4) improvements in 20-m speed occurred before-PHV but the effect was otherwise unclear with moderate to large individual differences. These data reflect specific responses to training interventions in girls of different biological maturity, while highlighting individual responses to HIIT interventions. This can assist practitioners in providing effective training prescription.

The use of big data to transform care delivery is rapidly becoming a reality. To deliver on the promise of value-based care, providers must know the key drivers of wellness at the patient and community levels, as well as understand resource constraints and opportunities to improve efficiency in the healthcare system itself. Data are the linchpin. By gathering the right data and finding innovative ways to glean knowledge, we can improve clinical care, advance the health of our communities, improve the lives of our patients, and operate more efficiently. At Carolinas HealthCare System-one of the nation's largest healthcare systems, with nearly 12 million patient encounters annually at more than 900 care locations-we have made substantial investments to establish a centralized data and analytics infrastructure that is transforming the way we deliver care across the continuum. Although the impetus and vision for our program have evolved over the past decade, our efforts coalesced into a strategic, centralized initiative with the launch of the Dickson Advanced Analytics (DA2) group in 2012. DA2 has yielded significant gains in our ability to use data, not only for reporting purposes and understanding our business but also for predicting outcomes and informing action.While these efforts have been successful, the path has not been easy. Effectively harnessing big data requires navigating myriad technological, cultural, operational, and other hurdles. Building a program that is feasible, effective, and sustainable takes concerted effort and a rigorous process of continuous self-evaluation and strategic adaptation.