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Background Oncofetal splice variants of extracellular matrix (ECM) proteins present a unique group of target antigens for the immunotherapy of pediatric cancers. However, limited data is available if these splice variants can be targeted with T cells expressing chimeric antigen receptors (CARs).Methods To determine the expression of the oncofetal version of tenascin C (TNC) encoding the C domain (C.TNC) in pediatric brain and solid tumors, we used quantitative reverse transcription PCR and immunohistochemistry. Genetically modified T cells were generated from human peripheral blood mononuclear cells and evaluated in vitro and in vivo.Results We demonstrate that C.TNC is expressed on a protein level in pediatric tumors, including diffuse intrinsic pontine glioma, osteosarcoma, rhabdomyosarcoma, and Ewing sarcoma. We generate C.TNC-CAR T cells and establish that these recognize and kill C.TNC-positive tumor cells. However, their antitumor activity in vivo is limited. To improve the effector function of C.TNC-CAR T cells, we design a leucine zipper-based chimeric cytokine receptor that activates interleukin-18 signaling pathways (Zip18R). Expression of Zip18R in C.TNC-CAR T cells improves their ability to secrete cytokines and expand in repeat stimulation assays. C.TNC-CAR.Zip18R T cells also have significantly greater antitumor activity in vivo compared with unmodified C.TNC-CAR T cells.Conclusions Our study identifies the C domain of the ECM protein TNC as a promising CAR T-cell therapy for pediatric solid tumors and brain tumors. While we focus here on pediatric cancer, our work has relevance to a broad range of adult cancers that express C.TNC.

Abstract Background The Covid-19 pandemic has exacerbated pre-existing inequalities and increased adversity and challenges for vulnerable and marginalised communities worldwide. In the UK, the Voluntary Community and Social Enterprise (VCSE) sector play a vital role in supporting the health and wellbeing of people who are marginalised or experiencing multiple complex needs. However, only a small number of studies have focused on the impact that Covid-19 had on the VCSE sector. Methods As part of a Health Inequalities Impact Assessment (HIIA), we conducted qualitative focus groups with staff and volunteers from five organisations to examine short, medium and longer-term impacts of Covid-19 upon the VCSE sector in Northern England. Nine online focus groups were conducted between March and July 2021. Findings Focus group transcripts were analysed using Framework Analysis and yielded three central themes: (1) exacerbation of pre-existing inequalities, adversity and challenges for vulnerable and marginalised populations; (2) the ‘price’ of being flexible, innovative and agile for VCSE staff and volunteers; and (3) the voluntary sector as a ‘lifeline’ - organisational pride and resilience. Conclusions While the voluntary sector ‘adapted at pace’ to provide support during Covid-19 and in its continued aftermath, this resilience has potentially come at the cost of workforce and volunteer wellbeing, compounded by political obstacles and chronic shortage in funding and support. The VCSE sector has a vital role to play in the post-lockdown ‘levelling up’ agenda. The expertise, capacity and resilience of VCSE organisations, and their ability to respond to Covid-19, should be celebrated, recognised and supported adequately to maintain its resilience. To not do so threatens the sector’s sustainability and risks jeopardising attempts to involve the sector in addressing the social determinants of health.

The goal of this study is to develop and validate a lightweight, interpretable machine learning (ML) classifier to identify opioid overdoses in emergency medical services (EMS) records. We conducted a comparative assessment of three feature engineering approaches designed for use with unstructured narrative data. Opioid overdose annotations were provided by two harm reduction paramedics and two supporting annotators trained to reliably match expert annotations. Candidate feature engineering techniques included term frequency-inverse document frequency (TF-IDF), a highly performant approach to concept vectorization, and a custom approach based on the count of empirically-identified keywords. Each feature set was trained using four model architectures: generalized linear model (GLM), Naïve Bayes, neural network, and Extreme Gradient Boost (XGBoost). Ensembles of trained models were also evaluated. The custom feature models were also assessed for variable importance to aid interpretation. Models trained using TF-IDF feature engineering ranged from AUROC = 0.59 (95% CI: 0.53-0.66) for the Naïve Bayes to AUROC = 0.76 (95% CI: 0.71-0.81) for the neural network. Models trained using concept vectorization features ranged from AUROC = 0.83 (95% 0.78-0.88)for the Naïve Bayes to AUROC = 0.89 (95% CI: 0.85-0.94) for the ensemble. Models trained using custom features were the most performant, with benchmarks ranging from AUROC = 0.92 (95% CI: 0.88-0.95) with the GLM to 0.93 (95% CI: 0.90-0.96) for the ensemble. The custom features model achieved positive predictive values (PPV) ranging for 80 to 100%, which represent substantial improvements over previously published EMS encounter opioid overdose classifiers. The application of this approach to county EMS data can productively inform local and targeted harm reduction initiatives.

BackgroundHIV disproportionately affects sexual minority men (SMM) in the United States. ObjectiveWe sought to determine past HIV postexposure prophylaxis (PEP) use and current and prior pre-exposure prophylaxis (PrEP) use among a web-based sample of cisgender and transgender men who have sex with men. MethodsIn 2019, HIV-negative and unknown status SMM (n=63,015) were recruited via geosocial networking apps, social media, and other web-based venues to participate in a brief eligibility screening survey. Individuals were asked about past PEP use and current and prior PrEP use. We examined associations of demographics, socioeconomic indicators, and recent club drug use with PEP and PrEP use, as well as the association between past PEP use and current and prior PrEP use using generalized linear models and multinomial logistic regression. Statistical significance was considered at P

Objectives The purpose of this study was to conduct a methodological review of research on the effects of conflicts of interest (COIs) in research contexts.Design Methodological review.Data sources Ovid.Eligibility criteria Studies published between 1986 and 2021 conducting quantitative assessments of relationships between industry funding or COI and four target outcomes: positive study results, methodological biases, reporting quality and results–conclusions concordance.Data extraction and synthesis We assessed key facets of study design: our primary analysis identified whether studies stratified industry funding or COI variables by magnitude (ie, number of COI or disbursement amount), type (employment, travel fees, speaking fees) or if they assessed dichotomous variables (ie, conflict present or absent). Secondary analyses focused on target outcomes and available effects measures.Results Of the 167 articles included in this study, a substantial majority (98.2%) evaluated the effects of industry sponsorship. None evaluated associations between funding magnitude and outcomes of interest. Seven studies (4.3%) stratified industry funding based on the mechanism of disbursement or funder relationship to product (manufacturer or competitor). A fifth of the articles (19.8%) assessed the effects of author COI on target outcomes. None evaluated COI magnitude, and three studies (9.1%) stratified COI by disbursement type and/or reporting practices. Participation of an industry-employed author showed the most consistent effect on favourability of results across studies.Conclusions Substantial evidence demonstrates that industry funding and COI can bias biomedical research. Evidence-based policies are essential for mitigating the risks associated with COI. Although most policies stratify guidelines for managing COI, differentiating COIs based on the type of relationship or monetary value, this review shows that the available research has generally not been designed to assess the differential risks of COI types or magnitudes. Targeted research is necessary to establish an evidence base that can effectively inform policy to manage COI.

During influenza A virus (IAV) entry, the hemagglutinin (HA) protein is triggered by endosomal low pH to undergo irreversible structural changes that mediate membrane fusion. HA proteins from different isolates vary in the pH at which they become activated in endosomes or become irreversible inactivated if exposed to extracellular acid. Little is known about extracellular pH in the upper respiratory tracts of mammals, how pH may shift during IAV infection, and its impact on replication of viruses that vary in HA activation pH. Here, we inoculated DBA/2J mice intranasally with A/TN/1-560/2009 (H1N1) (activation pH 5.5) or a mutant containing the destabilizing mutation HA1-Y17H (pH 6.0). We measured the kinetics of extracellular pH during infection using an optical pH-sensitive microsensor probe placed in the naris, nasal sinus, soft palate, and trachea. We also measured intracellular pH of single-cell suspensions of live, primary lung epithelial cells with various wavelength pH-sensitive dyes localized to cell membranes, cytosol, endosomes, secretory vesicles, microtubules, and lysosomes. Infection with either virus decreased extracellular pH and increased intracellular pH. Peak host immune responses were observed at 2 days post infection (DPI) and peak pH changes at 5 DPI. Extracellular and intracellular pH returned to baseline by 7 DPI in mice infected with HA1-Y17H and was restored later in wildtype-infected. Overall, IAV infection altered respiratory tract pH, which in turn modulated replication efficiency. This suggests a virus-host pH feedback loop that may select for IAV strains containing HA proteins of optimal pH stability, which may be approximately pH 5.5 in mice but may differ in other species.

Recently, concerns have been raised over the potential impacts of commercial relationships on editorial practices in biomedical publishing. Specifically, it has been suggested that certain commercial relationships may make editors more open to publishing articles with author conflicts of interest (aCOI). Using a data set of 128,781 articles published in 159 journals, we evaluated the relationships among commercial publishing practices and reported author conflicts of interest. The 159 journals were grouped according to commercial biases (reprint services, advertising revenue, and ownership by a large commercial publishing firm). 30.6% (39,440) of articles were published in journals showing no evidence of evaluated commercial publishing relationships. 33.9% (43,630) were published in journals accepting advertising and reprint fees; 31.7% (40,887) in journals owned by large publishing firms; 1.2% (1,589) in journals accepting reprint fees only; and 2.5% (3,235) in journals accepting only advertising fees. Journals with commercial relationships were more likely to publish articles with aCOI (9.2% (92/1000) vs. 6.4% (64/1000), p = 0.024). In the multivariate analysis, only a journal's acceptance of reprint fees served as a significant predictor (OR = 2.81 at 95% CI, 1.5 to 8.6). Shared control estimation was used to evaluate the relationships between commercial publishing practices and aCOI frequency in total and by type. BCa-corrected mean difference effect sizes ranged from -1.0 to 6.1, and confirm findings indicating that accepting reprint fees may constitute the most significant commercial bias. The findings indicate that concerns over the influence of industry advertising in medical journals may be overstated, and that accepting fees for reprints may constitute the largest risk of bias for editorial decision-making.

The common arguments about potential student use of these technologies suggest artificial intelligence (AI)-assisted writing will damage student learning by shortcutting the writing process. There is a further worry that AI-based pedagogy will de-skill students by reducing writing to a mere editing practice. Additionally, some suggest that students will fail to do adequate background research, relying on dubious or non-existent AI-generated sources. Ultimately, these concerns indicate a view of AI-assisted writing where students can essentially parachute their way into the last stages of the writing process, skipping past considerable effort and learning. S. Scott Graham largely rejects these worries because they simply do not fit with his understanding of how writing works. At their core, each of these concerns is marked by a problematic commitment to a limited and linear process-based model of writing. Instead, he thinks writing instructors and scholars of writing studies can consider how their hard-won insights about diverse and variable writing practices might allow for robust writing pedagogies that engage, rather than prohibit, use of AI. He argues in this article that there is already a post-process pedagogy however he first discusses what exactly these new AI systems can do.

In this work, speed of sound in 2 phase mixture has been explored using CFD-DEM (Computational Fluid Dynamcis - Discrete Element Modelling). In this method volume averaged Navier Stokes, continuity and energy equations are solved for fluid. Particles are simulated as individual entities; their behaviour is captured by Newton's laws of motion and classical contact mechanics. Particle-fluid interaction is captured using drag laws given in literature. The speed of sound in a medium depends on physical properties. It has been found experimentally that speed of sound drops significantly in 2 phase mixture of fluidised particles because of its increased density relative to gas while maintaining its compressibility. Due to the high rate of heat transfer within 2 phase medium as given in Roy et al. (1990), it has been assumed that the fluidised gas-particle medium is isothermal. The similar phenomenon has been tried to be captured using CFD-DEM numerical simulation. The disturbance is introduced and fundamental frequency in the medium is noted to measure the speed of sound for e.g. organ pipe. It has been found that speed of sound is in agreement with the relationship given in Roy et al. (1990). Their assumption that the system is isothermal also appears to be valid.

OBJECTIVE: To examine determinants of maternal mortality and assess the effect of programmes aimed at increasing the number of births attended by health professionals in two districts in West Java, Indonesia. METHODS: We used informant networks to characterize all maternal deaths, and a capture-recapture method to estimate the total number of maternal deaths. Through a survey of recent births we counted all midwives practising in the two study districts. We used case-control analysis to examine determinants of maternal mortality, and cohort analysis to estimate overall maternal mortality ratios. FINDINGS: The overall maternal mortality ratio was 435 per 100 000 live births (95% confidence interval, CI: 376-498). Only 33% of women gave birth with assistance from a health professional, and among them, mortality was extremely high for those in the lowest wealth quartile range (2303 per 100 000) and remained very high for those in the lower middle and upper middle quartile ranges (1218 and 778 per 100 000, respectively). This is perhaps because the women, especially poor ones, may have sought help only once a serious complication had arisen. CONCLUSION: Achieving equitable coverage of all births by health professionals is still a distant goal in Indonesia, but even among women who receive professional care, maternal mortality ratios remain surprisingly high. This may reflect the limitations of home-based care. Phased introduction of fee exemption and transport incentives to enable all women to access skilled delivery care in health centres and emergency care in hospitals may be a feasible, sustainable way to reduce Indonesia's maternal mortality ratio.

[This corrects the article DOI: 10.1371/journal.pone.0154412.].

The glaucomas are a group of diseases characterized by optic nerve damage that together represent a leading cause of blindness in the human population and in domestic animals. Here we report a mutation in LTBP2 that causes primary congenital glaucoma (PCG) in domestic cats. We identified a spontaneous form of PCG in cats and established a breeding colony segregating for PCG consistent with fully penetrant, autosomal recessive inheritance of the trait. Elevated intraocular pressure, globe enlargement and elongated ciliary processes were consistently observed in all affected cats by 8 weeks of age. Varying degrees of optic nerve damage resulted by 6 months of age. Although subtle lens zonular instability was a common feature in this cohort, pronounced ectopia lentis was identified in less than 10% of cats examined. Thus, glaucoma in this pedigree is attributed to histologically confirmed arrest in the early post-natal development of the aqueous humor outflow pathways in the anterior segment of the eyes of affected animals. Using a candidate gene approach, significant linkage was established on cat chromosome B3 (LOD 18.38, θ = 0.00) using tightly linked short tandem repeat (STR) loci to the candidate gene, LTBP2. A 4 base-pair insertion was identified in exon 8 of LTBP2 in affected individuals that generates a frame shift that completely alters the downstream open reading frame and eliminates functional domains. Thus, we describe the first spontaneous and highly penetrant non-rodent model of PCG identifying a valuable animal model for primary glaucoma that closely resembles the human disease, providing valuable insights into mechanisms underlying the disease and a valuable animal model for testing therapies.

Structural and gene expression changes in the microvasculature of the human choroid occur during normal aging and age-related macular degeneration (AMD). In this study, we sought to determine the impact of aging and AMD on expression of the endothelial cell glycoprotein CD34. Sections from 58 human donor eyes were categorized as either young (under age 40), age-matched controls (> age 60 without AMD), or AMD affected (>age 60 with early AMD, geographic atrophy, or choroidal neovascularization). Dual labeling of sections with Ulex europaeus agglutinin-I lectin (UEA-I) and CD34 antibodies was performed, and the percentage of capillaries labeled with UEA-I but negative for anti-CD34 was determined. In addition, published databases of mouse and human retinal pigment epithelium-choroid were evaluated and CD34 expression compared between young and old eyes. Immunohistochemical studies revealed that while CD34 and UEA-I were colocalized in young eyes, there was variable loss of CD34 immunoreactivity in older donor eyes. While differences between normal aging and AMD were not significant, the percentage of CD34 negative capillaries in old eyes, compared to young eyes, was highly significant (p = 3.8×10(-6)). Endothelial cells in neovascular membranes were invariably CD34 positive. Published databases show either a significant decrease in Cd34 (mouse) or a trend toward decreased CD34 (human) in aging. These findings suggest that UEA-I and endogenous alkaline phosphatase activity are more consistent markers of aging endothelial cells in the choroid, and suggest a possible mechanism for the increased inflammatory milieu in the aging choroid.

In traumatic brain injury (TBI), a diversity of brain resident and peripherally derived myeloid cells have the potential to worsen damage and/or to assist in healing. We define the heterogeneity of microglia and macrophage phenotypes during TBI in wild-type (WT) mice and Ccr2-/- mice, which lack macrophage influx following TBI and are resistant to brain damage. We use unbiased single-cell RNA sequencing methods to uncover 25 microglia, monocyte/macrophage, and dendritic cell subsets in acute TBI and normal brains. We find alterations in transcriptional profiles of microglia subsets in Ccr2-/- TBI mice compared to WT TBI mice indicating that infiltrating monocytes/macrophages influence microglia activation to promote a type I IFN response. Preclinical pharmacological blockade of hCCR2 after injury reduces expression of IFN-responsive gene, Irf7, and improves outcomes. These data extend our understanding of myeloid cell diversity and crosstalk in brain trauma and identify therapeutic targets in myeloid subsets.

Regular HIV testing is an essential component of the HIV prevention and care cascade. Sexual minority males (SMM) account for most new HIV infections in the US and testing rates among SMM vary substantially across the lifespan. Research has largely overlooked the developmental context of HIV testing. The current study compared correlates of HIV testing among adolescents (aged 13-17; n = 1,641), emerging adults (aged 18-29; n = 50,483), early adults (aged 30-39; n = 25,830), middle adults (aged 40-64; n = 25,326), and late adults (65 and older; n = 1,452) who were recruited online. Overall, HIV testing rates were lowest among adolescent SMM. Having condomless anal sex in the past 3-months was a consistentpredictor of HIV testing across all age cohorts.The association between relationship status and frequency of HIV testing varied across ages. Being in a non-monogamous relationship (versus single) was associated with more frequent HIV testing among adolescent and emerging adult SMM, while being in a monogamous relationship (versus single) was associated with lower odds of HIV testing among early, middle, and late adults., Author(s): Trey V. Dellucci [sup.1] [sup.2] , S. Scott Jones [sup.1] , Christel Adhemar [sup.1] , Sarah W. Feldstein Ewing [sup.3] , Travis I. Lovejoy [sup.4] [sup.5] , Tyrel J. [...]