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Abstract Background Scaling is typically discussed as a way to amplify or expand a health innovation. However, there is limited knowledge about the specific techniques that can enhance access to or improve the quality of innovations, aiming to increase their positive impacts for the public good. We sought to identify, compare, and contrast scaling frameworks to advance the science and practice of scaling. Methods Using a scoping review we asked: 1) What are the attributes of scaling frameworks for innovations that support health outcomes? and 2) What are the similarities and differences of these attributes? Inclusion criteria were 1) primary studies or review articles, 2) a primary focus on scaling innovations for health and social care, 3) articles that developed a framework, and 4) articles were concerned with a health outcome. Starting from an umbrella review, we identified relevant studies and extracted data about the characteristics of the articles, attributes of framework development, attributes of framework components, transferability, and the framework’s underlying ethical lens. Grey literature was included through expert consultation. Data were summarized using frequencies and qualitative description. Results From 94 potentially eligible articles, we identified 9 unique frameworks and included 4 additional frameworks from the grey literature, resulting in a total of 13 frameworks. Seven frameworks include a definition of scaling, and eight are designed for public health settings. Five of the frameworks were developed for the US/Canada/UK and Australia. Six of the lead authors’ primary institutional affiliation are from North America. Framework developers involved diverse stakeholders in a number of ways to develop their framework. Eight frameworks were developed, but not yet tested or applied, while the remaining frameworks were in the process of being applied or had already been applied to cases. All frameworks use a consequentialist-utilitarian ethical lens. Lastly, a comparison between frameworks found in the grey or published literature show important differences. Conclusion Much may be learned through further support for, and development of, scaling frameworks by primary authors affiliated with the Global South. Important aspects of framework development were identified, especially understanding the nuances of diverse stakeholder involvement in development.

Abstract Background Patients with kidney failure often lack robust evidence because they are excluded from randomized trials. Trial emulation provides an alternative approach to derive treatment effect estimates when randomized trials cannot be conducted. Critical questions about the comparative efficacy and safety of interventions in kidney failure are now being answered using this approach or parts of it. However, variations and inconsistencies in reporting cast doubt on the reliability and validity of effect estimates not derived from randomized trials. The aim of this methodological systematic review is to understand the extent to which the target study approach is used in kidney failure and the appropriateness of this approach. By identifying and evaluating studies that qualify as emulating a target trial, compared with studies that did not apply the principles. We aim to provide more specific methodological guidance to increase the clarity and reliability of reporting treatment effect estimates when running a trial in kidney failure is not feasible. Methods This protocol is developed in accordance with Preferred Reporting Items for Systematic reviews and Meta-Analyses Protocols (PRISMA-P) statement. MEDLINE, Embase, and reference lists (backwards citation chasing) will be searched up until 1st July 2023 and the search updated prior to publication to identify all studies evaluating patient outcomes in late-stage kidney disease and failure that use target trial emulation as the primary approach for analysis. Two authors (A. A., P. K.) will select articles based on title and abstract and then full text, with a third reviewer settling disagreements (J. P.). The prespecified variables will be extracted, and the risk of bias will be assessed by at least two authors (A. A., P. K., A. N.) using prespecified data forms. This will enable the determination of the robustness of the methodological quality of observational studies in using the whole or elements of the target trial approach. We will thereby assess their ability to reliably report treatment effect estimates. Discussion We will provide specific methodological recommendations on how to design target trials and model assumptions for emulation to get reliable treatment effect estimates for therapeutic interventions in kidney failure. Methodological systematic review registration Open Science Framework: Identifier https://doi.org/10.17605/OSF.IO/Z4Y29 .

Abstract Background and aims Several non-invasive tests for detecting bladder cancer (BC) are commercially available and are based on detecting small panels of BC-associated mutations and/or methylation changes in urine DNA. However, it is not clear which type of biomarker is best, or if a combination of the two is needed. In this study we address this question by taking a 23-gene mutation panel (GALEAS™ Bladder, GB) and testing if adding a panel of methylation markers improves the sensitivity of BC detection. Methods Twenty-three methylation markers were assessed in urine DNA by bisulphite conversion, multiplex PCR, and next generation sequencing in 118 randomly selected haematuria patients with pre-existing GB data (56 BCs and 62 non-BCs), split into training and test sets. We also analysed an additional 16 GB false-negative urine DNAs. Results The methylation panel detected bladder cancer in haematuria patients with 69% sensitivity at 96% specificity (test set results, 95% CIs 52-87% and 80-99%, respectively). Corresponding sensitivity and specificity for GB were 92% and 89%. Methylation and mutation markers were highly concordant in urine, with all GB false-negative samples also negative for methylation markers. Conclusions and limitations Our data show that, with a comprehensive mutation panel, any gains from adding methylation markers are, at best, marginal. It is likely that low tumour content is the commonest cause of false-negative urine test results. Our study does have a limited sample size and other methylation markers might behave differently to the those studied here.

Abstract Background Alternative polyadenylation (APA) affects most human genes and is recurrently dysregulated in all studied cancers. However, the mechanistic origins of this dysregulation are incompletely understood. Results We describe an unbiased analysis of molecular regulators of poly(A) site selection across The Cancer Genome Atlas and identify that colorectal adenocarcinoma is an outlier relative to all other cancer subtypes. This distinction arises from the frequent presence of loss-of-function APC mutations in colorectal adenocarcinoma, which are strongly associated with long 3′ UTR expression relative to tumors lacking APC mutations. APC knockout similarly dysregulates APA in human colon organoids. By mining previously published APC eCLIP data, we show that APC preferentially binds G- and C-rich motifs just upstream of proximal poly(A) sites. Lastly, we find that reduced APC expression is associated with APA dysregulation in tumor types lacking recurrent APC mutations. Conclusions As APC has been previously identified as an RNA-binding protein that preferentially binds 3′ UTRs during mouse neurogenesis, our results suggest that APC promotes proximal poly(A) site use and that APC loss and altered expression contribute to pervasive APA dysregulation in cancers.

Abstract Immune modulation and desensitization is a growing field of research and clinical investigation that requires precise delivery of antigens to immune system cells. Nanoparticles (NPs) have emerged as excellent candidates for antigen delivery, particularly in immune desensitization applications. NP-encapsulated protein antigens enable the delivery of protein and co-encapsulated adjuvant to antigen-presenting cells without systemic exposure and allergic response. Here, we show a method for producing poly(lactide-co-glycolide) (PLG) NPs in an efficient, high-yield, and large-scale inhomogeneous precipitation process. The process enables the production of compositionally complex PLG NPs containing protein while also incorporating DNA and E. coli phospholipids as integral adjuvants in the NP vehicle. Orally delivered PLG NPs activate the murine immune system, and encapsulated peanut allergen protein elicits approximately 10-fold lower levels of basophil activation than does unencapsulated protein in basophils isolated from peanut-allergic patients. This efficacy and safety evidence makes these PLG NPs excellent candidates for clinical applications.

Abstract Robotic locomotion in unstructured terrain demands an agile, adaptive, and energy-efficient architecture. To traverse such terrains, legged robots use rigid electromagnetic motors and sensorized drivetrains to adapt to the environment actively. These systems struggle to compete with animals that excel through their agile and effortless motion in natural environments. We propose a bio-inspired musculoskeletal leg architecture driven by antagonistic pairs of electrohydraulic artificial muscles. Our leg is mounted on a boom arm and can adaptively hop on varying terrain in an energy-efficient yet agile manner. It can also detect obstacles through capacitive self-sensing. The leg performs powerful and agile gait motions beyond 5 Hz and high jumps up to 40 % of the leg height. Our leg’s tunable stiffness and inherent adaptability allow it to hop over grass, sand, gravel, pebbles, and large rocks using only open-loop force control. The electrohydraulic leg features a low cost of transport (0.73), and while squatting, it consumes only a fraction of the energy (1.2 %) compared to its conventional electromagnetic counterpart. Its agile, adaptive, and energy-efficient properties would open a roadmap toward a new class of musculoskeletal robots for versatile locomotion and operation in unstructured natural environments.

Abstract Background Deletions and partial losses of chromosome 7 (chr7) are frequent in acute myeloid leukemia (AML) and are linked to dismal outcome. However, the genomic landscape and prognostic impact of concomitant genetic aberrations remain incompletely understood. Methods To discover genetic lesions in adult AML patients with aberrations of chromosome 7 [abn(7)], 60 paired diagnostic/remission samples were investigated by whole-exome sequencing in the exploration cohort. Subsequently, a gene panel including 66 genes and a SNP backbone for copy-number variation detection was designed and applied to the remaining samples of the validation cohort. In total, 519 patients were investigated, of which 415 received intensive induction treatment, typically containing a combination of cytarabine and anthracyclines. Results In the exploration cohort, the most frequently mutated gene was TP53 (33%), followed by epigenetic regulators (DNMT3A, KMT2C, IDH2) and signaling genes (NRAS, PTPN11). Thirty percent of 519 patients harbored ≥ 1 mutation in genes located in commonly deleted regions of chr7—most frequently affecting KMT2C (16%) and EZH2 (10%). KMT2C mutations were often subclonal and enriched in patients with del(7q), de novo or core-binding factor AML (45%). Cancer cell fraction analysis and reconstruction of mutation acquisition identified TP53 mutations as mainly disease-initiating events, while del(7q) or −7 appeared as subclonal events in one-third of cases. Multivariable analysis identified five genetic lesions with significant prognostic impact in intensively treated AML patients with abn(7). Mutations in TP53 and PTPN11 (11%) showed the strongest association with worse overall survival (OS, TP53: hazard ratio [HR], 2.53 [95% CI 1.66–3.86]; P

Abstract Outer membrane vesicles (OMVs) produced by Gram-negative bacteria have key roles in cell envelope homeostasis, secretion, interbacterial communication, and pathogenesis. The facultative intracellular pathogen Salmonella Typhimurium increases OMV production inside the acidic vacuoles of host cells by changing expression of its outer membrane proteins and modifying the composition of lipid A. However, the molecular mechanisms that translate pH changes into OMV production are not completely understood. Here, we show that the outer membrane protein PagC promotes OMV production through pH-dependent interactions between its extracellular loops and surrounding lipopolysaccharide (LPS). Structural comparisons and mutational studies indicate that a pH-responsive amino acid motif in PagC extracellular loops, containing PagC-specific histidine residues, is crucial for OMV formation. Molecular dynamics simulations suggest that protonation of histidine residues leads to changes in the structure and flexibility of PagC extracellular loops and their interactions with the surrounding LPS, altering membrane curvature. Consistent with that hypothesis, mimicking acidic pH by mutating those histidine residues to lysine increases OMV production. Thus, our findings reveal a mechanism for sensing and responding to environmental pH and for control of membrane dynamics by outer membrane proteins.

Abstract The rodent-borne Andes virus (ANDV) causes a severe disease in humans. We developed an ANDV mRNA vaccine based on the M segment of the viral genome, either with regular uridine (U-mRNA) or N1-methylpseudouridine (m1Ψ-mRNA). Female mice immunized by m1Ψ-mRNA developed slightly greater germinal center (GC) responses than U-mRNA-immunized mice. Single cell RNA and BCR sequencing of the GC B cells revealed similar levels of activation, except an additional cluster of cells exhibiting interferon response in animals vaccinated with U-mRNA but not m1Ψ-mRNA. Similar immunoglobulin class-switching and somatic hypermutations were observed in response to the vaccines. Female Syrian hamsters were immunized via a prime–boost regimen with two doses of each vaccine. The titers of glycoprotein-binding antibodies were greater for U-mRNA construct than for m1Ψ-mRNA construct; however, the titers of ANDV-neutralizing antibodies were similar. Vaccinated animals were challenged with a lethal dose of ANDV, along with a naïve control group. All control animals and two animals vaccinated with a lower dose of m1Ψ-mRNA succumbed to infection whereas other vaccinated animals survived without evidence of virus replication. The data demonstrate the development of a protective vaccine against ANDV and the lack of a substantial effect of m1Ψ modification on immunogenicity and protection in rodents.

Abstract Remarkable advances in protocol development have been achieved to manufacture insulin-secreting islets from human pluripotent stem cells (hPSCs). Distinct from current approaches, we devised a tunable strategy to generate islet spheroids enriched for major islet cell types by incorporating PDX1+ cell budding morphogenesis into staged differentiation. In this process that appears to mimic normal islet morphogenesis, the differentiating islet spheroids organize with endocrine cells that are intermingled or arranged in a core-mantle architecture, accompanied with functional heterogeneity. Through in vitro modelling of human pancreas development, we illustrate the importance of PDX1 and the requirement for EphB3/4 signaling in eliciting cell budding morphogenesis. Using this new approach, we model Mitchell-Riley syndrome with RFX6 knockout hPSCs illustrating unexpected morphogenesis defects in the differentiation towards islet cells. The tunable differentiation system and stem cell-derived islet models described in this work may facilitate addressing fundamental questions in islet biology and probing human pancreas diseases.

Background Unfavorable health‐related social needs (HRSNs) have the potential to worsen health and well‐being and drive health disparities. Its associations with cardiovascular health (CVH), assessed by Life's Essential 8, have not been comprehensively examined among US adults. Methods and Results We used the National Health and Nutrition Examination Survey 2011 to March 2020 data for adults aged ≥20 years. We grouped Life's Essential 8 scores as low (0–49), moderate (50–79), and high (80–100) CVH. We identified 8 unfavorable HRSNs and assigned a value of 1 for the unfavorable status of each. The number of unfavorable HRSNs was summed and ranged from 0 to 8, with higher numbers indicating more unfavorable HRSNs. We used multivariable linear and multinomial logistic regression to examine the association between HRSNs and CVH. A total of 14 947 participants were included (n=7340 male [49.3%]; mean [SE] age, 46.4 [0.35] years). A higher number of unfavorable HRSNs were associated with worse CVH: comparing adults with unfavorable HRSNs of 1–2, 3–4, and ≥5 to those with none, the fully adjusted prevalence ratios (95% CI) for low CVH were 1.42 (1.17–1.73), 2.11 (1.69–2.63), and 2.42 (1.90–3.08), respectively. The corresponding prevalence ratios (95% CI) for high CVH were 0.77 (0.68–0.87), 0.58 (0.49–0.67), and 0.46 (0.38–0.55). The associations were consistent across subgroups and in sensitivity analyses. Conclusions There was a graded association between unfavorable HRSNs and a higher prevalence of low CVH or lower prevalence of high CVH. Public health interventions targeting HRSNs might reduce health disparities and promote CVH.

ABSTRACT Introduction Gestational diabetes mellitus (GDM) continues to increase particularly for non‐European women. This study aimed to identify and quantify risk factors for women diagnosed with gestational diabetes in New Zealand to identify women at higher risk. Methods A national dataset of 601,166 eligible women who had ≥ 1 birth in New Zealand between January 2001 and December 2010 identified 11,459 women with gestational diabetes of whom 11,447 were randomly matched with 57,235 control women for age and year of delivery. Results Adjusted odds ratios (95% CI) showed higher odds of gestational diabetes for Asian (3.60, 3.39–3.82), Pacific (2.76, 2.57–2.96) and Māori (1.23, 1.15–1.31) women compared with European/Other women. Women most economically disadvantaged (1.44, 1.34–1.56), not registered with a lead maternity carer (1.16, 1.04–1.30) and those identified as smokers (1.20, 1.11–1.31) were more likely than control women to develop gestational diabetes. In contrast, women residing in rural (0.83, 0.77–0.88) and remote areas (0.68, 0.60–0.77) were less likely to develop gestational diabetes compared with women living in urban areas, and similarly for non‐New Zealand resident women (0.78, 0.72–0.85) compared with resident women. Conclusions Women who were diagnosed with gestational diabetes were more likely to be non‐European, economically disadvantaged, residing in urban areas, unregistered with a lead maternity carer and more likely to smoke. In addition to universal screening for pre‐existing diabetes, all women at risk of gestational diabetes should be identified and supported to undertake to a 75 g glucose challenge test between 24 and 28 weeks.

ABSTRACT Diffuse panbronchiolitis (DPB) is a rare, idiopathic inflammatory disease primarily diagnosed in East Asian populations. DPB is characterized by diffuse pulmonary lesions, inflammation of the respiratory bronchioles, and bacterial infections of the airway. Historically, sputum cultures reveal Pseudomonas aeruginosa in 22% of DPB patients, increasing to 60% after 4 years from disease onset. Although DPB patients have a known susceptibility to respiratory P. aeruginosa infections, as is observed in other chronic lung diseases such as cystic fibrosis (CF), the characterization of DPB P. aeruginosa strains is limited. In this study, we characterized 24 strains obtained from a cohort of DPB patients for traits previously associated with virulence, including growth, motility, antibiotic susceptibility, lipopolysaccharide structure, and genomic diversity. Our cohort of DPB P. aeruginosa strains exhibits considerable genomic variability when compared with isolates from people with cystic fibrosis chronically colonized with P. aeruginosa and acute P. aeruginosa infection isolates. Similar to CF, DPB P. aeruginosa strains produce a diverse array of modified lipid A structures. Antibiotic susceptibility testing revealed increased resistance to erythromycin, a representative agent of the macrolide antibiotics used to manage DPB patients. Differences in the O-antigen type among P. aeruginosa strains collected from these different backgrounds were also observed. Ultimately, the characterization of DPB P. aeruginosa strains highlights several unique qualities of P. aeruginosa strains collected from chronically diseased airways, underscoring the challenges in treating DPB, CF, and other obstructive respiratory disease patients with P. aeruginosa infections.IMPORTANCEDiffuse panbronchiolitis (DPB), a chronic lung disease characterized by persistent P. aeruginosa infection, serves as an informative comparator to more common chronic lung diseases, such as cystic fibrosis (CF). This study aimed to better address the interplay between P. aeruginosa and chronically compromised airway environments through the examination of DPB P. aeruginosa strains, as existing literature regarding DPB is limited to case reports, case series, and clinical treatment guidelines. The evaluation of these features in the context of DPB, in tandem with prevailing knowledge of P. aeruginosa strains collected from more common chronic lung diseases (e.g., CF), can aid in the development of more effective strategies to combat respiratory P. aeruginosa infections in patients with chronic lung diseases.

Hypoxic microenvironments induce widespread metabolic changes that have been shown to be critical in regulating innate and adaptive immune responses. Hypoxia-induced changes include the generation of extracellular adenosine followed by subsequent signaling through adenosine receptors on immune cells. This evolutionarily conserved “hypoxia-adenosinergic” pathway of hypoxia → extracellular adenosine → adenosine receptor signaling has been shown to be critical in limiting and redirecting T cell responses including in tumor microenvironments and the gut mucosa. However, the question of whether hypoxic microenvironments are involved in the development of B cell responses has remained unexplored until recently. The discovery that germinal centers (GC), the anatomic site in which B cells undergo secondary diversification and affinity maturation, develop a hypoxic microenvironment has sparked new interest in how this evolutionarily conserved pathway affects antibody responses. In this review we will summarize what is known about hypoxia-adenosinergic microenvironments in lymphocyte development and ongoing immune responses. Specific focus will be placed on new developments regarding the role of the hypoxia-adenosinergic pathway in regulating GC development and humoral immunity.

The precise control of soft and continuum robots requires knowledge of their shape, which has, in contrast to classical rigid robots, infinite degrees of freedom. To partially reconstruct the shape, proprioceptive techniques use built‐in sensors, resulting in inaccurate results and increased fabrication complexity. Exteroceptive methods so far rely on expensive tracking systems with reflective markers placed on all components, which are infeasible for deformable robots interacting with the environment due to marker occlusion and damage. Here, a regression approach is presented for three‐dimensional key point estimation using a convolutional neural network. The proposed approach uses data‐driven supervised learning and is capable of online markerless estimation during inference. Two images of a robotic system are captured simultaneously at 25 Hz from different perspectives and fed to the network, which returns for each pair the parameterized key point or piecewise constant curvature shape representations. The proposed approach outperforms markerless state‐of‐the‐art methods by a maximum of 4.5% in estimation accuracy while being more robust and requiring no prior knowledge of the shape. Online evaluations on two types of soft robotic arms and a soft robotic fish demonstrate the method's accuracy and versatility on highly deformable systems.

The Xerces Blue (Glaucopsyche xerces) is considered to be the first butterfly to become extinct in historical times. It was notable for its chalky lavender wings with conspicuous white spots on the ventral wings. The last individuals were collected in their restricted habitat, in the dunes near the Presidio military base in San Francisco, in 1941. We sequenced the genomes of four 80- to 100-year-old Xerces Blue, and seven historical and one modern specimens of its closest relative, the Silvery Blue (Glaucopsyche lygdamus). We compared these to a novel annotated genome of the Green-Underside Blue (Glaucopsyche alexis). Phylogenetic relationships inferred from complete mitochondrial genomes indicate that Xerces Blue was a distinct species that diverged from the Silvery Blue lineage at least 850,000 years ago. Using nuclear genomes, both species experienced population growth during the Eemian interglacial period, but the Xerces Blue decreased to a very low effective population size subsequently, a trend opposite to that observed in the Silvery Blue. Runs of homozygosity and deleterious load in the former were significantly greater than in the later, suggesting a higher incidence of inbreeding. These signals of population decline observed in Xerces Blue could be used to identify and monitor other insects threatened by human activities, whose extinction patterns are still not well known.

Background Although current guidelines recommend implantable cardioverter‐defibrillator (ICD) placement in survivors of out‐of‐hospital cardiac arrest, contemporary data on secondary‐prevention ICDs in survivors of out‐of‐hospital cardiac arrest remain limited. Methods and Results Using 2013 to 2019 CARES (Cardiac Arrest Registry to Enhance Survival) linked to Medicare, we identified 3226 patients aged ≥65 years with an initial shockable rhythm who survived to discharge without severe neurological disability. Multivariable hierarchical regression models were used to examine the association between patient variables and ICD placement and quantify hospital variation in ICD implantation. The mean age was 72.2 years, 23.5% were women, 10% were Black individuals, and 4% were Hispanic individuals. Overall, 997 (30.9%) patients received an ICD before discharge, 1266 (39.2%) at 90 days, and 1287 (39.9%) within 6 months. Older age (≥85 years), female sex, history of diabetes, calendar year, and presentation with acute myocardial infarction were associated with lower odds of ICD implantation, but race or ethnicity was not associated with ICD implantation. Among 297 hospitals, the median proportion of survivors receiving ICD at discharge was 28.6% (interquartile range, 20%–50%). The relative odds of ICD implantation varied by 62% across hospitals (median odds ratio, 1.62 [95% CI, 1.38–1.82]) after adjusting for case mix. Conclusions Fewer than 1 in 3 survivors of out‐of‐hospital cardiac arrest due to a shockable rhythm received a secondary‐prevention ICD before discharge. Although patient variables were associated with ICD implantation, there was no difference by race or ethnicity. Even after adjusting for patient case mix, ICD implantation varied markedly across hospitals.

Background: Inequities in stroke outcomes have existed for decades, and the COVID-19 pandemic amplified these inequities. Objectives: This study examined the association between social vulnerability and all-cause mortality among Medicare beneficiaries hospitalized with acute ischemic stroke (AIS) during COVID-19 pandemic periods. Methods: We analyzed data on Medicare fee-for-service beneficiaries aged ≥65 years hospitalized with AIS between April 1, 2020, and December 31, 2021 (followed until December 31, 2023) merged with county-level data from the 2020 Centers for Disease Control and Prevention/Agency for Toxic Substances and Disease Registry’s Social Vulnerability Index (SVI). We used a Cox proportional hazard model to examine the association between SVI quartile and all-cause mortality. Results: Among 176,123 Medicare fee-for-service beneficiaries with AIS, 29.9% resided in the most vulnerable counties (SVI quartile 4), while 14.9% resided in counties with least social vulnerability (SVI quartile 1). AIS Medicare beneficiaries living in the most vulnerable counties had the highest proportions of adults aged 65 to 74 years, non-Hispanic Black or Hispanic, severe stroke at admission, a history of COVID-19, and more prevalent comorbidities. Compared to those living in least vulnerable counties, AIS Medicare beneficiaries living in most vulnerable counties had significantly higher all-cause mortality (adjusted HR: 1.11, 95% CI: 1.08-1.14). The pattern of association was largely consistent in subgroup analyses by age group, sex, and race and ethnicity. Conclusions: Higher social vulnerability levels were associated with increased all-cause mortality among AIS Medicare beneficiaries. To improve outcomes and address disparities, it may be important to focus efforts toward addressing social vulnerability.

Abstract In this work we demonstrate a miniaturised imaging system based around a time-gated SPAD array operating in a “chip-on-tip” manner. Two versions of the system are demonstrated, each measuring 23 mm × 23 mm × 28 mm with differing fields of view and working distances. Initial tests demonstrate contrast between materials in widefield fluorescence imaging (WFLIm) mode, with frame rates of > 2 Hz achievable. Following this, WFLIm images of autofluorescence in ovine lung tissue are obtained at frame rates of ~ 1 Hz. Finally, the ability of the second system to perform simultaneous WFLIm and time of flight (aka Flourescence Lifetime Imaging Distance and Ranging, FLImDAR) is also tested. This shows that the system is capable of 4 mm resolution of object separation when tested on 3D printed samples. It is further demonstrated as being able to perform scene reconstruction on autofluorescent lung tissue. This system is, to date, the smallest chip on tip WFLIm system published, and is the first demonstration of the FLImDAR technique in a compact, portable system.

Abstract Mammalian inner ear hair cell loss leads to permanent hearing and balance dysfunction. In contrast to the cochlea, vestibular hair cells of the murine utricle have some regenerative capacity. Whether human utricular hair cells regenerate in vivo remains unknown. Here we procured live, mature utricles from organ donors and vestibular schwannoma patients, and present a validated single-cell transcriptomic atlas at unprecedented resolution. We describe markers of 13 sensory and non-sensory cell types, with partial overlap and correlation between transcriptomes of human and mouse hair cells and supporting cells. We further uncover transcriptomes unique to hair cell precursors, which are unexpectedly 14-fold more abundant in vestibular schwannoma utricles, demonstrating the existence of ongoing regeneration in humans. Lastly, supporting cell-to-hair cell trajectory analysis revealed 5 distinct patterns of dynamic gene expression and associated pathways, including Wnt and IGF-1 signaling. Our dataset constitutes a foundational resource, accessible via a web-based interface, serving to advance knowledge of the normal and diseased human inner ear.

Abstract Classical metabolomic and new metabolic network methods were used to study the developmental features of autism spectrum disorder (ASD) in newborns (n = 205) and 5-year-old children (n = 53). Eighty percent of the metabolic impact in ASD was caused by 14 shared biochemical pathways that led to decreased anti-inflammatory and antioxidant defenses, and to increased physiologic stress molecules like lactate, glycerol, cholesterol, and ceramides. CIRCOS plots and a new metabolic network parameter, $$\dot{{{\boldsymbol{V}}}}\!$$ V ° net , revealed differences in both the kind and degree of network connectivity. Of 50 biochemical pathways and 450 polar and lipid metabolites examined, the developmental regulation of the purine network was most changed. Purine network hub analysis revealed a 17-fold reversal in typically developing children. This purine network reversal did not occur in ASD. These results revealed previously unknown metabolic phenotypes, identified new developmental states of the metabolic correlation network, and underscored the role of mitochondrial functional changes, purine metabolism, and purinergic signaling in autism spectrum disorder.

Abstract Animal activity reflects behavioral decisions that depend upon environmental context. Prior studies typically estimated activity distributions within few areas, which has limited quantitative assessment of activity changes across environmental gradients. We examined relationships between two response variables, activity level (fraction of each day spent active) and pattern (distribution of activity across a diel cycle) of white-tailed deer (Odocoileus virginianus), with four predictors—deer density, anthropogenic development, and food availability from woody twigs and agriculture. We estimated activity levels and patterns with cameras in 48 different 10.36-km2 landscapes across three larger regions. Activity levels increased with greater building density, likely due to heightened anthropogenic disturbance, but did not vary with food availability. In contrast, activity patterns responded to an interaction between twigs and agriculture, consistent with a functional response in habitat use. When agricultural land was limited, greater woody twig density was associated with reduced activity during night and evening. When agricultural land was plentiful, greater woody twig density was associated with more pronounced activity during night and evening. The region with the highest activity level also experienced the most deer-vehicle collisions. We highlight how studies of spatial variation in activity expand ecological insights on context-dependent constraints that affect wildlife behavior.

Abstract Bromodomain and extra-terminal domain (BET) proteins are therapeutic targets in several cancers including the most common malignant adult brain tumor glioblastoma (GBM). Multiple small molecule inhibitors of BET proteins have been utilized in preclinical and clinical studies. Unfortunately, BET inhibitors have not shown efficacy in clinical trials enrolling GBM patients. One possible reason for this may stem from resistance mechanisms that arise after prolonged treatment within a clinical setting. However, the mechanisms and timeframe of resistance to BET inhibitors in GBM is not known. To identify the temporal order of resistance mechanisms in GBM we performed quantitative proteomics using multiplex-inhibitor bead mass spectrometry and demonstrated that intrinsic resistance to BET inhibitors in GBM treatment occurs rapidly within hours and involves the fibroblast growth factor receptor 1 (FGFR1) protein. Additionally, small molecule inhibition of BET proteins and FGFR1 simultaneously induces synergy in reducing GBM tumor growth in vitro and in vivo. Further, FGFR1 knockdown synergizes with BET inhibitor mediated reduction of GBM cell proliferation. Collectively, our studies suggest that co-targeting BET and FGFR1 may dampen resistance mechanisms to yield a clinical response in GBM.

Abstract Background Randomised controlled trials (RCTs) aim to estimate the causal effect of one or more interventions relative to a control. One type of outcome that can be of interest in an RCT is an ordinal outcome, which is useful to answer clinical questions regarding complex and evolving patient states. The target parameter of interest for an ordinal outcome depends on the research question and the assumptions the analyst is willing to make. This review aimed to provide an overview of how ordinal outcomes have been used and analysed in RCTs. Methods The review included RCTs with an ordinal primary or secondary outcome published between 2017 and 2022 in four highly ranked medical journals (the British Medical Journal, New England Journal of Medicine, The Lancet, and the Journal of the American Medical Association) identified through PubMed. Details regarding the study setting, design, the target parameter, and statistical methods used to analyse the ordinal outcome were extracted. Results The search identified 309 studies, of which 144 were eligible for inclusion. The most used target parameter was an odds ratio, reported in 78 (54%) studies. The ordinal outcome was dichotomised for analysis in 47 ( $$33\%$$ 33 % ) studies, and the most common statistical model used to analyse the ordinal outcome on the full ordinal scale was the proportional odds model (64 [ $$44\%$$ 44 % ] studies). Notably, 86 (60%) studies did not explicitly check or describe the robustness of the assumptions for the statistical method(s) used. Conclusions The results of this review indicate that in RCTs that use an ordinal outcome, there is variation in the target parameter and the analytical approaches used, with many dichotomising the ordinal outcome. Few studies provided assurance regarding the appropriateness of the assumptions and methods used to analyse the ordinal outcome. More guidance is needed to improve the transparent reporting of the analysis of ordinal outcomes in future trials.

Purpose: To assess the feasibility of a second-generation (44-channel) suprachoroidal retinal prosthesis for provision of functional vision in recipients with end-stage retinitis pigmentosa (RP) over 2.7 years. Design: Prospective, single-arm, unmasked interventional clinical trial. Participants: Four participants, with advanced RP and bare-light perception vision. Methods: The 44-channel suprachoroidal retinal prosthesis was implanted in the worse-seeing eye. Device stability, functionality, and adverse events were investigated at approximately 12-week intervals up to 140 weeks (2.7 years) postdevice activation. Main Outcome Measures: Serious adverse event (SAE) reporting, visual response outcomes, functional vision outcomes, and quality-of-life outcomes. Results: All 4 participants (aged 39–66 years, 3 males) were successfully implanted in 2018, and there were no device-related SAEs over the duration of the study. A mild postoperative subretinal hemorrhage was detected in 2 recipients, which cleared spontaneously within 2 weeks. OCT confirmed device stability and position under the macula. Improvements in localization abilities were demonstrated for all 4 participants in screen-based, tabletop, and orientation and mobility tasks. In addition, 3 of 4 participants recorded improvements in motion discrimination and 2 of 4 participants recorded substantial improvements in spatial discrimination and identification of tabletop objects. Participants reported their unsupervised use of the device included exploring new environments, detecting people, and safely navigating around obstacles. A positive effect of the implant on participants’ daily lives in their local environments was confirmed by an orientation and mobility assessor and participant self-report. Emotional well-being was not impacted by device implantation or usage. Conclusions: The completed clinical study demonstrates that the suprachoroidal prosthesis raises no safety concerns and provides improvements in functional vision, activities of daily living, and observer-rated quality of life. Financial Disclosure(s): Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

Abstract Declines in amphibian populations have led to greater research and monitoring of species of interest, including terrestrial salamanders. Several methods have been used to study salamanders, including artificial cover objects (ACOs). Artificial cover objects are easy to install and use, limit destruction to habitats, are easily reproducible both across and within studies, and can reduce variability between observers and sites by not relying on observer search time or effort, or on natural cover. However, different ACO designs and spacing may yield different capture rates, and thus different estimates of population densities. We examined the influence of ACO wood type, size, shape, and spacing arrays on the number of individually marked red‐backed salamanders (Plethodon cinereus) captured under ACOs. White pine (Pinus strobus) ACOs yielded the most (average of 14 across 3 replicates) individuals, while white ash (Fraxinus americana) ACOs had the least (average of 7 across 3 replicates). Larger ACOs had similar capture rates compared to equal sampled areas of smaller ACOs, but a single smaller ACO tended to have fewer individual salamander captures than a single large ACO. Our results suggested that high‐density ACO spacing arrays may increase capture probability but reduce the area uniquely sampled by each ACO. In contrast, low‐density ACO arrays may produce population estimates more comparable to other arrays. Future studies should consider monitoring goals and available sampling effort when choosing a spacing array. As ACO study design impacts salamander capture rates, a clear understanding of the effects of each element of ACO design is important when drawing inferences or making comparisons among studies.

Background: The World Health Organization has updated its classification of haematolymphoid tumours to the new 5th edition. The International Consensus Classification is another group that has developed a separate classification system with similar but conflicting approaches. This led to a debate among haematologists about which classification system is more appropriate for clinical use. This article examines and compares these contrasting and concurring classifications, and provides insight into the utility of each system. Methods: Journal articles pertaining to WHO-HAEM4R, WHO-HAEM5 and ICC were searched for using the MEDLINE database, from November 2022 to November 2023. Original research articles and reviews were selected to compile this review. Results: The classification systems share many similarities with minor name changes and subgrouping. There are few instances where diagnostic grouping differs between the classifications, which could have clinical implications regarding treatment and enrolment in clinical trials. Conclusion: There is universal agreement about the need for objective criteria for the classification of haematolymphoid tumours. As a result, there have been considerable strides in the classification with regard to morphologic, immunophenotypic, molecular and cytogenetic characterisation. It is reassuring to have a stepwise approach to diagnosis, allowing developing countries to make appropriate diagnoses. Contribution: Harmonisation is needed for a universal diagnostic system for the benefit of the patient. Both classification systems have merit and either could be adopted by individual institutions at present. The authors appeal for interim advocacy measures to assure access to the specific diagnostic investigations in poorly resourced societies, pending more permanent and sustainable access.

Many of the most highly conserved elements in the human genome are "poison exons," alternatively spliced exons that contain premature termination codons and permit post-transcriptional regulation of mRNA abundance through induction of nonsense-mediated mRNA decay (NMD). Poison exons are widely assumed to be highly conserved due to their presumed importance for organismal fitness, but this functional importance has never been tested in the context of a whole organism. Here, we report that a poison exon in Smndc1 is conserved across mammals and plants and plays a molecular autoregulatory function in both kingdoms. We generated mouse and A. thaliana models lacking this poison exon to find its loss leads to deregulation of SMNDC1 protein levels, pervasive alterations in mRNA processing, and organismal size restriction. Together, these models demonstrate the importance of poison exons for both molecular and organismal phenotypes that likely explain their extraordinary conservation.

Abstract Viral vectors and lipofection‐based gene therapies have dispersion‐dependent transduction/transfection profiles that thwart precise targeting. The study describes the development of focused close‐field gene electrotransfer (GET) technology, refining spatial control of gene expression. Integration of fluidics for precise delivery of “naked” plasmid deoxyribonucleic acid (DNA) in sucrose carrier within the focused electric field enables negative biasing of near‐field conductivity (“conductivity‐clamping”–CC), increasing the efficiency of plasma membrane molecular translocation. This enables titratable gene delivery with unprecedently low charge transfer. The clinic‐ready bionics‐derived CC‐GET device achieved neurotrophin‐encoding miniplasmid DNA delivery to the cochlea to promote auditory nerve regeneration; validated in deafened guinea pig and cat models, leading to improved central auditory tuning with bionics‐based hearing. The performance of CC‐GET is evaluated in the brain, an organ problematic for pulsed electric field‐based plasmid DNA delivery, due to high required currents causing Joule‐heating and damaging electroporation. Here CC‐GET enables safe precision targeting of gene expression. In the guinea pig, reporter expression is enabled in physiologically critical brainstem regions, and in the striatum (globus pallidus region) delivery of a red‐shifted channelrhodopsin and a genetically‐encoded Ca2+ sensor, achieved photoactivated neuromodulation relevant to the treatment of Parkinson's Disease and other focal brain disorders.

Background: Twin pregnancies are associated with higher risks of adverse neonatal outcomes compared to singleton pregnancies. The choice of delivery mode, when twin A presents cephalic, remains a subject of debate. In low- and middle-income countries, where healthcare resources are limited, the decision on the mode of delivery is even more critical. Objective: To evaluate the neonatal outcomes and the hospital costs of planned vaginal delivery compared to cesarean section (CS) in twin pregnancies with twin A presenting cephalic at Tenwek Hospital, Kenya. Study Design: This retrospective cohort study analyzed data from all twin deliveries at Tenwek Hospital, Kenya from, April 1, 2017, to March 30, 2023. Maternal data, mode of delivery, and neonatal data were collected from delivery logs, electronic health records, and neonatal records. Neonatal outcomes were a composite of either Appearance, Pulse, Grimace, Activity, and Respiration score less than seven at 5 minutes, neonatal intensive care unit admission, resuscitation, birth trauma, or neonatal complications, including death before discharge from the hospital. A logistic regression model was created to assess the impact of the planned mode of delivery on neonatal outcomes, controlling for antenatal care clinic visits, noncephalic presentation of twin B, and birth weight category. Results: The study included 177 twin deliveries: 129 (72.9%) were planned as vaginal deliveries and 48 (27.1%) were planned for CS. Among the planned vaginal deliveries, 66 (51.2%) experienced adverse outcomes, compared to 14 (29.2%) in the CS group (P=.009). Logistic regression showed that the odds of adverse outcomes were 0.35 times lower in the CS group compared to the planned vaginal delivery group (95% CI: 0.15–0.83; P=.017). The average total hospital costs for planned vaginal delivery were 104,608 Kenya Shillings (standard deviation 111,761) compared to 100,708 Kenya Shillings (standard deviation 75,468) for CS (P=.82). Conclusion: Planned cesarean deliveries in twin pregnancies with twin A presenting cephalic at Tenwek Hospital were associated with fewer adverse neonatal outcomes compared to planned vaginal deliveries. There was no significant difference in hospital costs. These findings raise the question of the safest mode of delivery for patients in a resource-constrained setting.

ObjectiveIn order to gain a better understanding of the individual and joint impact of testosterone and cortisol on behavior, the present study was developed to test the differences in each hormone alone and conjointly between perpetrators of IPV and non-violent controls.MethodPerpetrators of IPV on probation were compared to a control group of non-aggressive males from Hidalgo County in the Rio Grande Valley on baseline testosterone and cortisol, as well as several relevant questionnaires measuring aggression and trait anger. Differences in cortisol following exposure to a stressful event were also examined. Procedures included two laboratory visits consisting of questionnaires, a number of salivary testosterone and cortisol collections, and exposure to a stressor.ResultsPerpetrators had higher basal testosterone and post stressor cortisol levels than non- violent controls as well as a higher T/C ratio. In addition, trait anger moderated the relationship between both testosterone alone, and the testosterone/cortisol ratio and perpetration of IPV.ConclusionResults are consistent with the hypothesis that testosterone leads to antisocial behavior, including perpetration of violence. The results are also consistent with the dual hormone hypothesis, i.e., that testosterone and cortisol work together to jointly regulate social dominance and aggression. Both the increased freestanding testosterone and the increased cortisol following exposure to stress places these men at risk for perpetrating violence. Clinical implications are discussed.

Abstract A wide-field microscope with epi-fluorescence and selective plane illumination was combined with a single-photon avalanche diode (SPAD) array camera to enable live-cell fluorescence lifetime imaging (FLIM) using time-correlated single-photon counting (TCSPC). The camera sensor comprised of $$\text {192}\times \text {128}$$ 192 × 128 pixels, each integrating a single SPAD and a time-to-digital converter. Jointly, they produced a stream of single-photon images of photon arrival times with $$\approx \text {38 ps}$$ ≈ 38 ps accuracy. The photon arrival times were subject to systematic delays and nonlinearities, which were corrected by a Monte-Carlo algorithm. The SPAD camera was then applied to FLIM where histogramming the resulting photon arrival times in each pixel resulted in decays compatible with common data processing pipelines for fluorescence lifetime analysis. The capabilities of the TCSPC camera-based FLIM microscope were demonstrated by imaging living unicellular photosynthetic algae and artificial lipid vesicles. Epi-fluorescence illumination enabled rapid fluorescence lifetime imaging of living cells and selective-plane illumination enabled 3-dimensional FLIM of stationary samples.

Abstract Acquiring labeled datasets often incurs substantial costs primarily due to the requirement of expert human intervention to produce accurate and reliable class labels. In the modern data landscape, an overwhelming proportion of newly generated data is unlabeled. This paradigm is especially evident in domains such as fraud detection and datasets for credit card fraud detection. These types of data have their own difficulties associated with being highly class imbalanced, which poses its own challenges to machine learning and classification. Our research addresses these challenges by extensively evaluating a novel methodology for synthesizing class labels for highly imbalanced credit card fraud data. The methodology uses an autoencoder as its underlying learner to effectively learn from dataset features to produce an error metric for use in creating new binary class labels. The methodology aims to automatically produce new labels with minimal expert input. These class labels are then used to train supervised classifiers for fraud detection. Our empirical results show that the synthesized labels are of high enough quality to produce classifiers that significantly outperform a baseline learner comparison when using area under the precision-recall curve (AUPRC). We also present results of varying levels of positive-labeled instances and their effect on classifier performance. Results show that AUPRC performance improves as more instances are labeled positive and belong to the minority class. Our methodology thereby effectively addresses the concerns of high class imbalance in machine learning by creating new and effective class labels.

Abstract Background Autism spectrum disorder (ASD) and attention-deficit/hyperactivity disorder (ADHD) are neurodevelopmental disorders with overlapping behavioral features and genetic etiology. While brain cortical thickness (CTh) alterations have been reported in ASD and ADHD separately, the degree to which ASD and ADHD are associated with common and distinct patterns of CTh changes is unclear. Methods We searched PubMed, Web of Science, Embase, and Science Direct from inception to 8 December 2023 and included studies of cortical thickness comparing youth (age less than 18) with ASD or ADHD with typically developing controls (TDC). We conducted a comparative meta-analysis of vertex-based studies to identify common and distinct CTh alterations in ASD and ADHD. Results Twelve ASD datasets involving 458 individuals with ASD and 10 ADHD datasets involving 383 individuals with ADHD were included in the analysis. Compared to TDC, ASD showed increased CTh in bilateral superior frontal gyrus, left middle temporal gyrus, and right superior parietal lobule (SPL) and decreased CTh in right temporoparietal junction (TPJ). ADHD showed decreased CTh in bilateral precentral gyri, right postcentral gyrus, and right TPJ relative to TDC. Conjunction analysis showed both disorders shared reduced TPJ CTh located in default mode network (DMN). Comparative analyses indicated ASD had greater CTh in right SPL and TPJ located in dorsal attention network and thinner CTh in right TPJ located in ventral attention network than ADHD. Conclusions These results suggest shared thinner TPJ located in DMN is an overlapping neurobiological feature of ASD and ADHD. This alteration together with SPL alterations might be related to altered biological motion processing in ASD, while abnormalities in sensorimotor systems may contribute to behavioral control problems in ADHD. The disorder-specific thinner TPJ located in disparate attention networks provides novel insight into distinct symptoms of attentional deficits associated with the two neurodevelopmental disorders. Trial registration PROSPERO CRD42022370620. Registered on November 9, 2022.

Abstract Alternative polyadenylation (APA) is strikingly dysregulated in many cancers. Although global APA dysregulation is frequently associated with poor prognosis, the importance of most individual APA events is controversial simply because few have been functionally studied. Here, we address this gap by developing a CRISPR-Cas9-based screen to manipulate endogenous polyadenylation and systematically quantify how APA events contribute to tumor growth in vivo. Our screen reveals individual APA events that control mouse melanoma growth in an immunocompetent host, with concordant associations in clinical human cancer. For example, forced Atg7 3′ UTR lengthening in mouse melanoma suppresses ATG7 protein levels, slows tumor growth, and improves host survival; similarly, in clinical human melanoma, a long ATG7 3′ UTR is associated with significantly prolonged patient survival. Overall, our study provides an easily adaptable means to functionally dissect APA in physiological systems and directly quantifies the contributions of recurrent APA events to tumorigenic phenotypes.

Abstract Background Diabetes is a disease affecting over 500 million people globally due to insulin insufficiency or insensitivity. For individuals with type 1 diabetes, pancreatic islet transplantation can help regulate their blood glucose levels. However, the scarcity of cadaveric donor islets limits the number of people that could receive this therapy. To address this issue, human pluripotent stem cells offer a potentially unlimited source for generating insulin-producing cells through directed differentiation. Several protocols have been developed to make stem cell-derived insulin-producing cells. However, there is a lack of knowledge regarding the bioprocess parameters associated with these differentiation protocols and how they can be utilized to increase the cell yield. Methods We investigated various bioprocess parameters and quality target product profiles that may influence the differentiation pipeline using a seven-stage protocol in a scalable manner with CellSTACKs and vertical wheel bioreactors (PBS-Minis). Results Cells maintained > 80% viability through all stages of differentiation and appropriately expressed stage-specific markers. During the initial four stages leading up to the development of pancreatic progenitors, there was an increase in cell numbers. Following pancreatic progenitor stage, there was a gradual decrease in the percentage of proliferative cells, as determined by Ki67 positivity, and a significant loss of cells during the period of endocrine differentiation. By minimizing the occurrence of aggregate fusion, we were able to enhance cell yield during the later stages of differentiation. We suggest that glucose utilization and lactate production are cell quality attributes that should be considered during the characterization of insulin-producing cells derived from stem cells. Our findings also revealed a gradual metabolic shift from glycolysis, during the initial four stages of pancreatic progenitor formation, to oxidative phosphorylation later on during endocrine differentiation. Furthermore, the resulting insulin-producing cells exhibited a response to several secretagogues, including high glucose. Conclusion This study demonstrates process parameters such as glucose consumption and lactate production rates that may be used to facilitate the scalable manufacture of stem cell-derived insulin-producing cells.

While multi-agent frameworks can provide many advanced features, they often suffer from not being able to seamlessly interact with the outside world, e.g., with web-services or other multi-agent frameworks. This may be one factor that hinders a broader application of multi-agent systems in production systems. A possible solution to this problem is the combination of multi-agent systems with the concepts of micro-services and containerization, providing language-agnostic open interfaces, as well as encapsulation and modularity. In this paper, we propose an API and reference implementation that can be employed by multi-agent systems based on different languages and frameworks. Each agent component is encapsulated in a container and is accessed through its parent runtime platform, which takes care of aspects such as authentication, input validation, monitoring and other infrastructure tasks. Multiple runtime platforms can then be connected to form systems of distributed, heterogeneous multi-agent societies.

Abstract New generations of vaccines have been developed by encapsulating messenger ribonucleic acid (mRNA) in lipid nanoparticle (LNP) carriers. In addition to the physicochemical properties of LNPs, the encapsulation efficiency (EE) of mRNA in LNPs is a key factor to screen vaccine assembly assays. Fluorescent dyes with amplified signals upon binding with mRNA are at the core of developing assays to quantify EE. However, disregarding the temporal effects during the assay impacts the accuracy of the assay. Here, the kinetics of temporal decay in fluorescence intensity of dye‐RNA complex—in Ribogreen assay—are reported and shown how this dynamic process can be impeded in the presence of a nonionic surfactant. Further, the impact of this dynamic process on the calculated EE is studied. The corrections needed to accurately assay dynamic mRNA loading processes are presented.

BackgroundThis study unveils the intricate functional association between cyclic di-3’,5’-adenylic acid (c-di-AMP) signaling, cellular bioenergetics, and the regulation of lipopolysaccharide (LPS) profile in Porphyromonas gingivalis, a Gram-negative obligate anaerobe considered as a keystone pathogen involved in the pathogenesis of chronic periodontitis. Previous research has identified variations in P. gingivalis LPS profile as a major virulence factor, yet the underlying mechanism of its modulation has remained elusive.MethodsWe employed a comprehensive methodological approach, combining two mutants exhibiting varying levels of c-di-AMP compared to the wild type, alongside an optimized analytical methodology that combines conventional mass spectrometry techniques with a novel approach known as FLATn.ResultsWe demonstrate that c-di-AMP acts as a metabolic nexus, connecting bioenergetic status to nuanced shifts in fatty acid and glycosyl profiles within P. gingivalis LPS. Notably, the predicted regulator gene cdaR, serving as a potent regulator of c-di-AMP synthesis, was found essential for producing N-acetylgalactosamine and an unidentified glycolipid class associated with the LPS profile.ConclusionThe multifaceted roles of c-di-AMP in bacterial physiology are underscored, emphasizing its significance in orchestrating adaptive responses to stimuli. Furthermore, our findings illuminate the significance of LPS variations and c-di-AMP signaling in determining the biological activities and immunostimulatory potential of P. gingivalis LPS, promoting a pathoadaptive strategy. The study expands the understanding of c-di-AMP pathways in Gram-negative species, laying a foundation for future investigations into the mechanisms governing variations in LPS structure at the molecular level and their implications for host-pathogen interactions.

With high fatality and no cure, chronic wasting disease (CWD) has infected cervids in multiple regions, including the United States, Canada, Europe, and South Korea. Despite the rapid growth of literature on CWD, the full scope of its ecological, social, and economic impacts and the most effective and socially acceptable management strategies to mitigate the disease is unclear. Of 3008 initially identified published peer-reviewed papers, 134 were included in a final systematic literature review to synthesize the current knowledge on CWD transmission patterns, impacts, and the effectiveness of management interventions. The number of publications on CWD has increased steadily since 2000 with an average of six papers per year. Most papers were related to CWD prevalence (39 %), human behavior (33 %), CWD impacts (31 %), and management interventions (16 %). Environmental factors such as soil, water, and plants were identified as the most common transmission medium, with a higher prevalence rate among adult male cervids than females. Hunters showed a higher risk perception and were more likely to change hunting behavior due to CWD detection than non-hunters. Ecological impacts included the decreased survival rate accompanied by lower population growth, eventually leading to the decline of cervid populations. Culling was found to be an effective and widely implemented management strategy across countries, although it often was associated with public resistance. Despite potentially high negative economic impacts anticipated due to CWD, studies on this subject were limited. Sustained surveillance, ongoing research, and engagement of affected stakeholders will be essential for future disease control and management.

Cancer immune evasion contributes to checkpoint immunotherapy failure in many patients with metastatic cancers. The embryonic transcription factor DUX4 was recently characterized as a suppressor of interferon-γ signaling and antigen presentation that is aberrantly expressed in a small subset of primary tumors. Here, we report that DUX4 expression is a common feature of metastatic tumors, with ~10–50% of advanced bladder, breast, kidney, prostate, and skin cancers expressing DUX4. DUX4 expression is significantly associated with immune cell exclusion and decreased objective response to PD-L1 blockade in a large cohort of urothelial carcinoma patients. DUX4 expression is a significant predictor of survival even after accounting for tumor mutational burden and other molecular and clinical features in this cohort, with DUX4 expression associated with a median reduction in survival of over 1 year. Our data motivate future attempts to develop DUX4 as a biomarker and therapeutic target for checkpoint immunotherapy resistance.

Objective: Alström Syndrome (AS), caused by biallelic ALMS1 mutations, includes obesity with disproportionately severe insulin resistant diabetes, dyslipidemia, and fatty liver. Prior studies suggest that hyperphagia is accounted for by loss of ALMS1 function in hypothalamic neurones, whereas disproportionate metabolic complications may be due to impaired adipose tissue expandability. We tested this by comparing the metabolic effects of global and mesenchymal stem cell (MSC)-specific Alms1 knockout. Methods: Global Alms1 knockout (KO) mice were generated by crossing floxed Alms1 and CAG-Cre mice. A Pdgfrα-Cre driver was used to abrogate Alms1 function selectively in MSCs and their descendants, including preadipocytes. We combined metabolic phenotyping of global and Pdgfrα+ Alms1-KO mice on a 45% fat diet with measurements of body composition and food intake, and histological analysis of metabolic tissues. Results: Assessed on 45% fat diet to promote adipose expansion, global Alms1 KO caused hyperphagia, obesity, insulin resistance, dyslipidaemia, and fatty liver. Pdgfrα-cre driven KO of Alms1 (MSC KO) recapitulated insulin resistance, fatty liver, and dyslipidaemia in both sexes. Other phenotypes were sexually dimorphic: increased fat mass was only present in female Alms1 MSC KO mice. Hyperphagia was not evident in male Alms1 MSC KO mice, but was found in MSC KO females, despite no neuronal Pdgfrα expression. Conclusions: Mesenchymal deletion of Alms1 recapitulates metabolic features of AS, including fatty liver. This confirms a key role for Alms1 in the adipose lineage, where its loss is sufficient to cause systemic metabolic effects and damage to remote organs. Hyperphagia in females may depend on Alms1 deficiency in oligodendrocyte precursor cells rather than neurones. AS should be regarded as a forme fruste of lipodystrophy.

Abstract Background Gestational diabetes mellitus increases the risk of developing type 2 diabetes. The aim of this study is to compare cardiometabolic and renal outcomes for all women in New Zealand with gestational diabetes (2001–2010) with women without diabetes, 10–20 years following delivery. Methods A retrospective cohort study, utilizing a national dataset providing information for all women who gave birth between 1 January 2001 and 31 December 2010 (n = 604 398). Adolescent girls

Pseudomonas aeruginosa (Pa) is an opportunistic bacterial pathogen responsible for severe hospital acquired infections in immunocompromised and elderly individuals. Emergence of increasingly drug resistant strains and the absence of a broad-spectrum prophylactic vaccine against both T3SA+ (type III secretion apparatus) and ExlA+/T3SA- Pa strains worsen the situation in a post-pandemic world. Thus, we formulated a candidate subunit vaccine (called ExlA/L-PaF/BECC/ME) against both Pa types. This bivalent vaccine was generated by combining the C-terminal active moiety of exolysin A (ExlA) produced by non-T3SA Pa strains with our T3SA-based vaccine platform, L-PaF, in an oil-in-water emulsion. The ExlA/L-PaF in ME (MedImmune emulsion) was then mixed with BECC438b, an engineered lipid A analogue and a TLR4 agonist. This formulation was administered intranasally (IN) to young and elderly mice to determine its potency across a diverse age-range. The elderly mice were used to mimic the infection seen in elderly humans, who are more susceptible to serious Pa disease compared to their young adult counterparts. After Pa infection, mice immunized with ExlA/L-PaF/BECC/ME displayed a T cell-mediated adaptive response while PBS-vaccinated mice experienced a rapid onset inflammatory response. Important genes and pathways were observed, which give rise to an anti-Pa immune response. Thus, this vaccine has the potential to protect aged individuals in our population from serious Pa infection.

The collaborative trans-European dimension of Modernism engenders particularly three huge potentials – especially exploring the developments beyond east and West dichotomies: first, a scope of incredibly rich and diverse modern cultural heritage, second the vital realm and diversity of historic protagonists as well as of current actors, topics, and formats and, third, its fruitful relevance and diverse perspectives for contemporary challenges and opportunities.

We performed a thorough systematic review of published literature to determine potential links between human health impacts and naled, a registered adult mosquito control product (adulticide), and its major degradate, dichlorvos (DDVP). A search query was performed on 8 September 2023, capturing all articles published up to that date on the Scopus and PubMed databases. Inclusion criteria were the presence of either pesticide and a measured or modeled human health outcome or risk. The search string resulted in 382 articles; however, 354 articles were excluded, resulting in only 28 articles that met the inclusion criteria. The studies that directly relate to aerosolized ultra-low volume (ULV) mosquito control applications did not report any associated deleterious human health outcomes. Results from the reviewed papers displayed no negative health effects or led to inconclusive results. No studies showed adverse health effects from aerial ULV applications for mosquito management. Our findings are congruent with the United States Environmental Protection Agency and Centers for Disease Control and Prevention recommendations that aerial applications of naled, following label parameters, do not pose an adverse risk exposure to humans, wildlife, and the environment.