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Neonatal-perinatal medicine (NPM) fellowship training aims to educate fellows and enable them to move into successful independent neonatal practice. To reach this goal, every fellow must acquire a wide range of clinical, scholarly, and professional skills that require assessment and evaluation by the fellowship program. The assessment and evaluation of fellow progression and promotion must rely on a rigorous educational framework. The goal of this article is to illustrate the process of NPM fellow assessment and evaluation, introduce the educational framework utilized, describe challenges inherent to the process, and discuss opportunities to improve how NPM fellows are evaluated., Competing Interests: Competing interests: The authors declare no competing interests., (© 2025. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Variability in space use among conspecifics can emerge from foraging strategies that track available resources, especially in riverscapes that promote high synchrony between prey pulses and consumers. Projected changes in riverscape hydrological regimes due to water management and climate change accentuate the need to understand the natural variability in animal space use and its implications for population dynamics and ecosystem function. Here, we used long-term tracking of Common Snook (Centropomus undecimalis) movement and trophic dynamics in the Shark River, Everglades National Park from 2012 to 2023 to test how specialization in the space use of individuals (i.e., E adj ) changes seasonally, how it is influenced by yearly hydrological conditions, and its relationship to the between individual trophic niche. Snook exhibited seasonal variability in space use, with maximum individual specialization (high dissimilarity) in the wet season. The degree of individual specialization increased over the years in association with greater marsh flooding duration, which produced important subsidies. Also, there were threshold responses of individual space use specialization as a function of floodplain conditions. Greater specialization in space use results in a decrease in snook trophic niche size. These results show how hydrological regimes in riverscapes influence individual specialization of resource use (both space and prey), providing insight into how forecasted hydroclimatic scenarios may shape habitat selection processes and the trophic dynamics of mobile consumers., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethical approval: The Florida International University Institution Animal Care and Use Committee (IACUC-18-002-CR01) approved the project’s sampling and animal handling protocol. Project activities were conducted under the Everglades National Park research permit EVER-2022-SCI-0056. This study is reported following ARRIVE guidelines., (© 2025. The Author(s).)

Marine foundation species are increasingly impacted by anthropogenic stressors, driving a loss of diversity within these critical habitats. Prior studies suggest that species diversity within mussel beds has declined precipitously in southern California, USA, but it is unclear whether a similar loss has occurred farther north. Here, we resurvey a mussel bed community in northern California first sampled in 1941 to evaluate changes in diversity after 78 years. More broadly, we explore the value and potential challenges of using imperfect historical data to assess community changes. Our 2019 survey documented 90 species/taxa across 10 phyla. The majority of species (~ 72%) were common to all replicate plots, suggesting that variation in species diversity over small spatial scales was unlikely to mask temporal changes. In contrast to results from southern California, we observed no decline in species diversity between timepoints. However, there were shifts in species composition, with an increase in the abundance of southern species and a decrease in northern species, consistent with warming observed at a nearby shoreline site. Overall, our findings are an encouraging sign for the health of this mussel bed community in northern California and illustrate how non-traditional data can contribute to assessments of long-term ecological change., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Objective: The Downes score is a neonatal examination scoring tool frequently used to guide initiation of CPAP, but its ability to predict the need for surfactant has not been assessed. We assessed the extent to which the Downes score predicts the receipt of surfactant., Study Design: We calculated a simplified Downes score from nursing admission data for infants (≤ 2000 grams, and ≥ 25 weeks' gestation) admitted on CPAP to a highly resourced level III NICU, to assess the predictive value for the receipt of surfactant., Results: Fifty-three (31.5%) out of 168 infants admitted on CPAP received surfactant. A simplified Downes score of ≥ 4 predicted the receipt of surfactant with 90.6% sensitivity, 52.2% specificity, 46.6% positive predictive value, 92.3% negative predictive value, and 64.3% accuracy., Conclusion: The high sensitivity and negative predictive value suggest utility for using the Downes score to help guide clinical decision making regarding surfactant therapy., Competing Interests: Competing interests: The authors declare no competing interests. Ethics approval: The University of Vermont Institutional Review Board determined that this study was not human subjects research. This analysis of existing data was performed in accordance with the Declaration of Helsinki., (© 2024. The Author(s).)

Intraspecific variation is important for species' long-term persistence in changing environments. Conservation strategies targeting intraspecific variation often rely on the identification of management or policy units below the species level based on biological differences among populations. To identify management units, this paper examines intraspecific divergence of Lake Trout (Salvelinus namaycush) in Great Slave Lake (GSL), Canada, using low-coverage whole-genome sequencing data. Specifically, we evaluate genetic differentiation and assess the relationship with morphological, mitochondrial, and putatively adaptive divergence. We show that at least three and up to five genetically distinct Lake Trout populations co-occur in GSL and exhibit differences in spatial distribution and body size, with signatures of selection. However, admixture was widespread (60% of the fish) and population structure was shallow (average F ST  = 0.022). These findings highlight that, even in the era of whole genome sequencing, identifying discrete management units to implement conservation efforts and policy can remain challenging in systems where gene flow among genetically and ecologically distinct populations is ubiquitous. To give more recognition to this complexity, management efforts should also focus on the area where adaptive variation is evident, and evolutionary acts are at play, to better protect species' resilience and adaptive potential in some natural systems., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Background: Hepatocellular carcinoma (HCC) is a leading cause of cancer-related deaths worldwide, often linked to chronic inflammation. Our study aimed to probe inflammation pathways at the genetic level and pinpoint biomarkers linked to HCC patient survival., Methods: We analyzed gene transcriptome data from 246 resectable stage I and II HCC patients from The Cancer Genome Atlas (TCGA). After selecting 917 inflammation-related genes (IRGs), we identified 104 differentially expressed genes (DEGs) through differential expression analysis. Two significant prognostic DEGs, S100A9 and PBK, were identified using LASSO and Cox regression, forming the basis of a risk score model. We conducted functional enrichment and immune landscape analyses, validated our findings on 170 patients from the GSE14520 dataset, and performed mutational analysis using TCGA somatic mutation data., Results: We analyzed 296 samples (246 HCC, 50 normal liver), showing significant survival differences between high and low-risk groups based on our risk score model. Functional enrichment analysis unveiled inflammation-associated pathways. Validation using the GSE14520 dataset confirmed our risk score's predictive ability, and we explored clinical correlations., Conclusion: Our study delineates inflammation-related genomic changes in HCC, unveiling prognostic biomarkers with potential therapeutic implications. These findings deepen our understanding of HCC molecular mechanisms and may guide personalized therapeutic approaches, ultimately improving patient outcomes., Competing Interests: Declarations. Financial support: This research has not received external funding. Conflict of interest: Anwaar Saeed reports a leadership role with Autem therapeutics, Exelixis, KAHR medical and Bristol-Myers Squibb; consulting or advisory board role with AstraZeneca, Bristol-Myers Squibb, Merck, Exelixis, Pfizer, Xilio therapeutics, Taiho, Amgen, Autem therapeutics, KAHR medical, and Daiichi Sankyo; institutional research funding from AstraZeneca, Bristol-Myers Squibb, Merck, Clovis, Exelixis, Actuate therapeutics, Incyte Corporation, Daiichi Sankyo, Five prime therapeutics, Amgen, Innovent biologics, Dragonfly therapeutics, Oxford Biotherapeutics, Arcus therapeutics, and KAHR medical; and participation as a data safety monitoring board chair for Arcus therapeutics. The remaining authors have no relevant financial interests to disclose., (© 2024. The Author(s).)

Leptin is an adipokine associated with obesity and with hypertension in animal models. Whether leptin is associated with hypertension independent of obesity is unclear. Relative to White adults, Black adults have higher circulating leptin concentration. As such, leptin may mediate some of the excess burden of incident hypertension among Black adults. REGARDS enrolled 30,239 adults aged ≥45 years from 48 US states in 2003-07. Baseline leptin was measured in a sex- and race-stratified sample of 4400 participants. Modified Poisson regression estimated relative risk (RR) of incident hypertension (new ≥140/≥90 mmHg threshold or use of antihypertensives) per SD of log-transformed leptin, stratified by obesity (BMI of 30 kg/m 2 ). Inverse odds ratio weighting estimated the % mediation by leptin of the excess hypertension RR among Black relative to White participants. Among the 1821 participants without prevalent hypertension, 35% developed incident hypertension. Obesity modified the relationship between leptin and incident hypertension (P-interaction 0.006) such that higher leptin was associated with greater hypertension risk in the crude model among those with BMI < 30 kg/m 2 , but not those with BMI ≥ 30 kg/m 2 . This was fully attenuated when adjusting for anthropometric measures. In the crude model, Black adults had a 52% greater risk of incident hypertension. Leptin did not significantly mediate this disparity. In this national U.S. sample, leptin was associated with incident hypertension among non-obese but not obese adults. Future investigations should focus on the effect of weight modification on incident hypertension among non-obese adults with elevated leptin., Competing Interests: Competing interests: DL Long, PhD received investigator-initiated research support from Amgen, Inc. for work unrelated to this manuscript. Ethical approval: Study methods and protocols were approved by the institutional review board of the University of Alabama at Birmingham, with protocols cited in the previous design paper. All participants provided written informed consent. No identifiable participant information is published herein. All methods were carried out in accordance with relevant guidelines and regulations., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

Mastitis is an inflammatory disease affecting mammary tissues caused by bacterial infection that negatively affects milk quality and quantity. S-Allylmercaptocysteine (SAMC), a sulfur compound in aged garlic extract (AGE), suppresses lipopolysaccharide (LPS)-induced inflammation in mouse models and cell cultures. However, the mechanisms underlying this anti-inflammatory effect remain unclear. In this study, we demonstrated that oral administration of AGE suppressed the LPS-induced immune response in a mastitis mouse model and that SAMC inhibited LPS-induced interleukin-6 production and nuclear factor κB p65 subunit activation in HC11 mammary epithelial cells. Global phosphoproteomic analysis revealed that SAMC treatment downregulated 910 of the 1,304 phosphorylation sites upregulated by LPS stimulation in mammary cells, including those associated with toll-like receptor 4 (TLR4) signaling. Additionally, SAMC decreased the phosphorylation of 26 proteins involved in pre-mRNA splicing, particularly the U2 small nuclear ribonucleoprotein complex. Furthermore, we found that SAMC increased the production of the myeloid differentiation factor 88 short form (MyD88-S), an alternatively spliced form of MyD88 that negatively regulates TLR4 signaling. These findings suggest that SAMC inhibits TLR4-mediated inflammation via alternative pre-mRNA splicing, thus promoting MyD88-S production in mammary epithelial cells. Therefore, SAMC may alleviate various inflammatory diseases, such as mastitis, by modulating immune responses., Competing Interests: Declarations. Competing interests: M.T., M.K., H.T., and J.I.S. are employed by Wakunaga Pharmaceutical Co., Ltd., and F.Q.Z. is employed by University of Vermont as listed in the affiliation section. The employment does not affect each researcher’s conceptualization, design, data collection, analysis, decision to publish, or preparation of the manuscript. This study revealed the new biological activity of SAMC in the experiment systems. The company and the University have no financial interest in this publication., (© 2024. The Author(s).)

We investigate the band structure of metal-dielectric photonic crystals comprising stacked organic semiconductor microcavities with silver metal mirrors incorporating crystal defects: individual unit cells with aperiodic dimensionality. Both transfer matrix simulation and experimental verification are performed to investigate the impact on the photonic band structure as a single cavity is varied in size. The resulting mid-gap defect states are shown to hybridize with a photonic band at certain resonant dimensions. The resonance of the defect cavity affects the transmittance of light through the device, disrupting or enhancing the coupling between otherwise resonant cavities. We outline potential applications for defect engineering of these devices through controlled manipulation of the transmission spectrum., Competing Interests: Declarations. Competing interests: M.S.W. and D.A. are inventors on PCT Patent Application No. PCT/US21/71679 with the University of Vermont and State Agricultural College as applicant. This patent covers light emitting devices with coupled resonant photonic unit cells., (© 2024. The Author(s).)

This study examined the effectiveness of various electrical stimulation methods in alleviating symptoms and enhancing the quality of life for women with urinary incontinence. We conducted a systematic search of PubMed, Cochrane Library, PEDro, EMBASE, and ClinicalTrials.gov from inception to August 2024. Randomized controlled trials (RCTs) that meet following criteria were included, urinary continence in women, using various electric stimulation treatments and evaluated outcomes related to symptoms, quality of life. Thirty RCTs were subjected to risk of bias assessment, certainty of evidence, and network meta-analysis. Statistical analysis was performed using a random-effects model, with continuous variables expressed as standardized mean difference (SMD) and 95% confidence interval (CI). Percutaneous tibial stimulation (SMD - 1.86, 95% CI - 2.77 to - 0.96) and intravaginal stimulation (SMD - 0.97, 95% CI - 1.55 to - 0.38) significantly reduced symptom severity. Additionally, percutaneous tibial, intravaginal, transcutaneous tibial, and trans-sacral stimulations improved quality of life. Percutaneous tibial stimulation was the most effective, followed by intravaginal stimulation. Despite moderate to low confidence in the evidence, large-scale RCTs are needed to evaluate long-term benefits of these treatment., (© 2024. The Author(s).)

Neuroimaging plays a crucial role in understanding brain structure and function, but the lack of transparency, reproducibility, and reliability of findings is a significant obstacle for the field. To address these challenges, there are ongoing efforts to develop reporting checklists for neuroimaging studies to improve the reporting of fundamental aspects of study design and execution. In this review, we first define what we mean by a neuroimaging reporting checklist and then discuss how a reporting checklist can be developed and implemented. We consider the core values that should inform checklist design, including transparency, repeatability, data sharing, diversity, and supporting innovations. We then share experiences with currently available neuroimaging checklists. We review the motivation for creating checklists and whether checklists achieve their intended objectives, before proposing a development cycle for neuroimaging reporting checklists and describing each implementation step. We emphasize the importance of reporting checklists in enhancing the quality of data repositories and consortia, how they can support education and best practices, and how emerging computational methods, like artificial intelligence, can help checklist development and adherence. We also highlight the role that funding agencies and global collaborations can play in supporting the adoption of neuroimaging reporting checklists. We hope this review will encourage better adherence to available checklists and promote the development of new ones, and ultimately increase the quality, transparency, and reproducibility of neuroimaging research., (© 2024. The Author(s), under exclusive licence to American College of Neuropsychopharmacology.)

The fruit fly Drosophila melanogaster has emerged as a key model organism in neuroscience, in large part due to the concentration of collaboratively generated molecular, genetic and digital resources available for it. Here we complement the approximately 140,000 neuron FlyWire whole-brain connectome 1 with a systematic and hierarchical annotation of neuronal classes, cell types and developmental units (hemilineages). Of 8,453 annotated cell types, 3,643 were previously proposed in the partial hemibrain connectome 2 , and 4,581 are new types, mostly from brain regions outside the hemibrain subvolume. Although nearly all hemibrain neurons could be matched morphologically in FlyWire, about one-third of cell types proposed for the hemibrain could not be reliably reidentified. We therefore propose a new definition of cell type as groups of cells that are each quantitatively more similar to cells in a different brain than to any other cell in the same brain, and we validate this definition through joint analysis of FlyWire and hemibrain connectomes. Further analysis defined simple heuristics for the reliability of connections between brains, revealed broad stereotypy and occasional variability in neuron count and connectivity, and provided evidence for functional homeostasis in the mushroom body through adjustments of the absolute amount of excitatory input while maintaining the excitation/inhibition ratio. Our work defines a consensus cell type atlas for the fly brain and provides both an intellectual framework and open-source toolchain for brain-scale comparative connectomics., (© 2024. The Author(s).)

Connections between neurons can be mapped by acquiring and analysing electron microscopic brain images. In recent years, this approach has been applied to chunks of brains to reconstruct local connectivity maps that are highly informative 1-6 , but nevertheless inadequate for understanding brain function more globally. Here we present a neuronal wiring diagram of a whole brain containing 5 × 10 7 chemical synapses 7 between 139,255 neurons reconstructed from an adult female Drosophila melanogaster 8,9 . The resource also incorporates annotations of cell classes and types, nerves, hemilineages and predictions of neurotransmitter identities 10-12 . Data products are available for download, programmatic access and interactive browsing and have been made interoperable with other fly data resources. We derive a projectome-a map of projections between regions-from the connectome and report on tracing of synaptic pathways and the analysis of information flow from inputs (sensory and ascending neurons) to outputs (motor, endocrine and descending neurons) across both hemispheres and between the central brain and the optic lobes. Tracing from a subset of photoreceptors to descending motor pathways illustrates how structure can uncover putative circuit mechanisms underlying sensorimotor behaviours. The technologies and open ecosystem reported here set the stage for future large-scale connectome projects in other species., (© 2024. The Author(s).)

Visitors to Colter Bay Village in Grand Teton National Park were surveyed to elicit their evaluations of experimental outdoor lighting conditions. Luminaires capable of dimming and switching between two LED modules (white, blended red-white) were installed in street and parking areas. The blended red-white lamps consisted of 30 narrowband LED with a peak wavelength 623 nm and two 3000 K white LEDs. Similar "red" lamps were previously shown to reduce impacts to bats and insects. The white and red lamps were closely matched for luminance. Measured horizontal illuminance at survey locations had an interquartile range from 0.63 to 3.82 lx. The red lamps produced lower perceived brightness (V B2 (λ)), even after reflection off asphalt, yet survey participants expressed higher ratings for visual comfort and safety under red lighting. Surveys conducted earlier in the evening, with higher levels of predicted solar and measured horizontal illuminance, rated higher on visual comfort and safety, though these correlations were not as strong as the effect of lamp color. Streetlight ratings and support for lighting that protected natural resources were not contingent upon age or gender. Survey participants assessed red lighting as more protective of the environment. These results demonstrate that outdoor lighting designed to reduce ecological impacts can yield superior nocturnal experience for pedestrians., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Plant-mediated synthesis of nanoparticles is a sustainable approach that has gained widespread scientific acceptance due to its numerous benefits and applications. In this study, a zinc oxide-doped activated carbon (ZAC) derived from palm kernel shells (PKS) was synthesized via a bioreduction route using a water-based extract of Nymphaea lotus leaves as a reducing agent. The synthesized ZAC nanocomposites were characterized using microscopic (TEM, SEM) and spectroscopic (FTIR, EDS, XRD, and UV-Vis) analyses. The adsorptive properties of ZAC and efficiency in scavenging a phenothiazine derivative (methylene blue) from an aqueous solution were investigated. Results reveal that nano-scale ZAC particles were crystalline, exhibited irregular shapes, with an average size of 45 nm, and were highly dispersed. The optimum quantity adsorbed was 248 mg/g at a methylene blue concentration of 140 mg/L for 60 min using 0.02 g/100 mL of ZAC. Adsorption and kinetics data closely aligned with the Freundlich isotherm and the pseudo-second-order model, respectively indicating heterogeneous surface adsorption and chemisorption as the dominant mechanisms. The regeneration study of ZAC shows that over five cycles, thermal regeneration maintained high adsorption capacity with minimal decline and chemical regeneration significantly led to reduction in the adsorption capacity but solvent washing offered a balance between efficacy and structural preservation., (© 2024. The Author(s).)

The 2018 African swine fever (ASF) outbreak highlighted the importance of biosecurity in food production systems. Despite the significant economic impacts, the sociopsychological consequences on decision-making have been overlooked. Previous studies have focused on algebraic models and simulation-based models without considering the complex psychological and social factors that influence farmers' biosecurity behaviors and decision-making processes. This study aims to classify livestock producers into distinct subgroups based on their attitudes towards biosecurity. We conducted a survey presenting producers with three scenarios to assess their willingness to report suspected ASF cases, trust in government agencies, risk perception, biosecurity knowledge, willingness to purchase livestock insurance, motivation to invest in biosecurity, readiness to report suspected infections, and intention to contact a veterinarian. Using latent class analysis, we identified three distinct classes: Biosecurity Sceptics, Biosecurity Compliant, and Biosecurity Ultra-Compliant. Our results show that producer characteristics significantly influence biosecurity attitudes and class membership, with small-scale producers less likely to adopt ultra-compliant biosecurity practices. Attending at least one eradication program encouraged biosecurity compliance. This research informs the design of targeted food policy and risk communication strategies that account for attitudes of livestock producers to encourage biosecurity adoption and reduce the likelihood of Tier 1 disease incursion., (© 2024. The Author(s).)

Within the scope of this investigation, we carried out experiments to investigate the potential of the Vision Transformer (ViT) in the field of medical image analysis. The diagnosis of osteoporosis through inspection of X-ray radio-images is a substantial classification problem that we were able to address with the assistance of Vision Transformer models. In order to provide a basis for comparison, we conducted a parallel analysis in which we sought to solve the same problem by employing traditional convolutional neural networks (CNNs), which are well-known and commonly used techniques for the solution of image categorization issues. The findings of our research led us to conclude that ViT is capable of achieving superior outcomes compared to CNN. Furthermore, provided that methods have access to a sufficient quantity of training data, the probability increases that both methods arrive at more appropriate solutions to critical issues., (© 2024. The Author(s).)

We investigated the prevalence, incidence, and rates of pharmacological treatment of delayed ejaculation using the TriNetX Diamond Network. We included all men evaluated in the inpatient, outpatient, and emergency settings. Prevalence was determined by comparing the number of men diagnosed with delayed ejaculation to the entire population. Incidence was determined by comparing the number of men diagnosed with delayed ejaculation without a prior diagnosis to the overall population without a prior diagnosis. Rates of pharmacologic treatment were calculated by comparing the number of men who received a prescription to the total number of men with delayed ejaculation. Trends in prevalence and incidence were compared using six-month intervals, while trends in pharmacologic treatment were compared using one-year intervals. A total of 23,164 adult males were diagnosed with delayed ejaculation from 2013 to 2019. During the final six-month interval (July to December 2019), 2,747 of 16,496,744 men received a delayed ejaculation diagnosis, and 1,375 of 16,488,270 men without a prior diagnosis were diagnosed with delayed ejaculation. In 2019, only 916 of 4,733 (19.4%) men diagnosed with delayed ejaculation received any prescription, with the most common being testosterone (9.5%), bupropion (6.6%), and buspirone (2.3%). Prevalence, incidence and pharmacologic treatment all had increasing trends., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system and a leading cause of neurological disability in young adults. Clinical presentation and disease course are highly heterogeneous. Typically, disease progression occurs over time and is characterized by the gradual accumulation of disability. The risk of developing MS is driven by complex interactions between genetic and environmental factors, including the gut microbiome. How the commensal gut microbiota impacts disease severity and progression over time remains unknown. In a longitudinal study, disability status and associated clinical features in 58 MS patients were tracked over 4.2 ± 0.98 years, and the baseline fecal gut microbiome was characterized via 16S amplicon sequencing. Progressor status, defined as patients with an increase in Expanded Disability Status Scale (EDSS), were correlated with features of the gut microbiome to determine candidate microbiota associated with risk of MS disease progression. We found no overt differences in microbial community diversity and overall structure between MS patients exhibiting disease progression and non-progressors. However, a total of 41 bacterial species were associated with worsening disease, including a marked depletion in Akkermansia, Lachnospiraceae, and Oscillospiraceae, with an expansion of Alloprevotella, Prevotella-9, and Rhodospirillales. Analysis of the metabolic potential of the inferred metagenome from taxa associated with progression revealed enrichment in oxidative stress-inducing aerobic respiration at the expense of microbial vitamin K 2 production (linked to Akkermansia), and a depletion in SCFA metabolism (linked to Oscillospiraceae). Further, as a proof of principle, statistical modeling demonstrated that microbiota composition and clinical features were sufficient to predict disease progression. Additionally, we found that constipation, a frequent gastrointestinal comorbidity among MS patients, exhibited a divergent microbial signature compared with progressor status. These results demonstrate a proof of principle for the utility of the gut microbiome for predicting disease progression in MS in a small well-defined cohort. Further, analysis of the inferred metagenome suggested that oxidative stress, vitamin K 2 , and SCFAs are associated with progression, warranting future functional validation and mechanistic study., (© 2024. The Author(s).)

Metabolic disease is epidemiologically linked to severe complications upon influenza virus infection, thus vaccination is a priority in this high-risk population. Yet, vaccine responses are less effective in these same hosts. Here we examined how the timing of diet switching from a high-fat diet to a control diet affected influenza vaccine efficacy in diet-induced obese mice. Our results demonstrate that the systemic meta-inflammation generated by high-fat diet exposure limited T cell maturation to the memory compartment at the time of vaccination, impacting the recall of effector memory T cells upon viral challenge. This was not improved with a diet switch post-vaccination. However, the metabolic dysfunction of T cells was reversed if weight loss occurred 4 weeks before vaccination, restoring a functional recall response. This corresponded with changes in the systemic obesity-related biomarkers leptin and adiponectin, highlighting the systemic and specific effects of diet on influenza vaccine immunogenicity., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

Members of the family Trichomeriaceae, belonging to the Chaetothyriales order and the Ascomycota phylum, are known for their capability to inhabit hostile environments characterized by extreme temperatures, oligotrophic conditions, drought, or presence of toxic compounds. The genus Knufia encompasses many polyextremophilic species. In this report, the genomic and morphological features of the strain FJI-L2-BK-P2 presented, which was isolated from the Mars 2020 mission spacecraft assembly facility located at the Jet Propulsion Laboratory in Pasadena, California. The identification is based on sequence alignment for marker genes, multi-locus sequence analysis, and whole genome sequence phylogeny. The morphological features were studied using a diverse range of microscopic techniques (bright field, phase contrast, differential interference contrast and scanning electron microscopy). The phylogenetic marker genes of the strain FJI-L2-BK-P2 exhibited highest similarities with type strain of Knufia obscura (CBS 148926 T ) that was isolated from the gas tank of a car in Italy. To validate the species identity, whole genomes of both strains (FJI-L2-BK-P2 and CBS 148926 T ) were sequenced, annotated, and strain FJI-L2-BK-P2 was confirmed as K. obscura. The morphological analysis and description of the genomic characteristics of K. obscura FJI-L2-BK-P2 may contribute to refining the taxonomy of Knufia species. Key morphological features are reported in this K. obscura strain, resembling microsclerotia and chlamydospore-like propagules. These features known to be characteristic features in black fungi which could potentially facilitate their adaptation to harsh environments., (© 2024. The Author(s).)

Mutations of the human leucine-rich repeat kinase 2 (LRRK2) have been associated with both, idiopathic and familial Parkinson's disease (PD). Most of these pathogenic mutations are located in the kinase domain (KD) or GTPase domain of LRRK2. In this study we describe a mechanism in which protein kinase activity can be modulated by reversible oxidation or reduction, involving a unique pair of adjacent cysteines, the "CC" motif. Among all human protein kinases, only LRRK2 contains this "CC" motif (C2024 and C2025) in the Activation Segment (AS) of the kinase domain. In an approach combining site-directed mutagenesis, biochemical analyses, cell-based assays, and Gaussian accelerated Molecular Dynamics (GaMD) simulations we could attribute a role for each of those cysteines. We employed reducing and oxidizing agents with potential clinical relevance to investigate effects on kinase activity and microtubule docking. We find that each cysteine gives a distinct contribution: the first cysteine, C2024, is essential for LRRK2 protein kinase activity, while the adjacent cysteine, C2025, contributes significantly to redox sensitivity. Implementing thiolates (R-S - ) in GaMD simulations allowed us to analyse how each of the cysteines in the "CC" motif interacts with its surrounding residues depending on its oxidation state. From our studies we conclude that oxidizing agents can downregulate kinase activity of hyperactive LRRK2 PD mutations and may provide promising tools for therapeutic strategies., (© 2024. The Author(s).)

Motor neurons are the final common pathway 1 through which the brain controls movement of the body, forming the basic elements from which all movement is composed. Yet how a single motor neuron contributes to control during natural movement remains unclear. Here we anatomically and functionally characterize the individual roles of the motor neurons that control head movement in the fly, Drosophila melanogaster. Counterintuitively, we find that activity in a single motor neuron rotates the head in different directions, depending on the starting posture of the head, such that the head converges towards a pose determined by the identity of the stimulated motor neuron. A feedback model predicts that this convergent behaviour results from motor neuron drive interacting with proprioceptive feedback. We identify and genetically 2 suppress a single class of proprioceptive neuron 3 that changes the motor neuron-induced convergence as predicted by the feedback model. These data suggest a framework for how the brain controls movements: instead of directly generating movement in a given direction by activating a fixed set of motor neurons, the brain controls movements by adding bias to a continuing proprioceptive-motor loop., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

Global climate change and land use change underlie a need to develop new crop breeding strategies, and crop wild relatives (CWR) have become an important potential source of new genetic material to improve breeding efforts. Many recent approaches assume adaptive trait variation increases towards the relative environmental extremes of a species range, potentially missing valuable trait variation in more moderate or typical climates. Here, we leveraged distinct genotypes of wild chickpea (Cicer reticulatum) that differ in their relative climates from moderate to more extreme and perform targeted assessments of drought and heat tolerance. We found significance variation in ecophysiological function and stress tolerance between genotypes but contrary to expectations and current paradigms, it was individuals from more moderate climates that exhibited greater capacity for stress tolerance than individuals from warmer and drier climates. These results indicate that wild germplasm collection efforts to identify adaptive variation should include the full range of environmental conditions and habitats instead of only environmental extremes, and that doing so may significantly enhance the success of breeding programs broadly., (© 2024. The Author(s).)

We aim to clarify the relationship between low skeletal muscle mass and varying levels of adiposity and to identify the types of physical function impairments associated with sarcopenic obesity (SO). This study examined cross-sectional data from the National Health and Nutrition Examination Survey with whole-body dual-energy X-ray absorptiometry (DXA) scans. The data included age, gender, DXA-assessed body composition, and physical functional activity with performing daily tasks by questionnaire. We subdivided the data by body composition into a non-SO group and a SO group (ASMI 0-49.99% and FMI of 50-100%), after which the SO data were subdivided into three classes. A higher class indicated higher adiposity and lower muscle mass. The physical function impairment of the two groups was compared. Our study examined 7161 individuals, of which 4907 did not have SO and 2254 had SO, and their data were further divided into three classes (i.e., class I, 826 individuals; class II, 1300 individuals; and class III, 128 individuals). Significant differences in demographics and DXA parameters were identified between the non-SO and SO groups (P < 0.001); the individuals with SO were older, included more women, and exhibited high adiposity and less lean muscle mass. The individuals with class III SO exhibited greater differences and reported more difficulty in performing daily activities. The individuals with class III SO exhibited the most severe physical function impairment. Our study highlights the considerable difficulties encountered by individuals with SO in performing daily activities. Given this finding, customized rehabilitation strategies should be implemented to improve the quality of life of individuals with SO., (© 2024. The Author(s).)

Both food insecurity and home and wild food procurement (HWFP), including gardening, increased in many countries during the COVID-19 pandemic; yet little evidence has demonstrated what impact HWFP had on food security. Using data from a representative sample of nearly 1000 residents in the two most rural US states (Vermont and Maine) conducted via an online survey in Spring/Summer 2021, as well as matching techniques, we compare food security outcomes among households who did and did not participate in HWFP in the first year of the pandemic. Nearly 60% of respondents engaged in HWFP in some way during the first year of the pandemic, with food insecure households more likely to do HWFP. Furthermore, HWFP early in the COVID-19 pandemic is associated with improved food security in the 9-12 months later, though these improvements were primarily associated with newly, not chronically, food insecure households. Newly and chronically food insecure households were more likely to want to continue these activities in the future, but also exhibited greater barriers to land access and costs associated with these activities. These results suggest that HWFP may provide food security improvements for certain households that utilize them, especially during crisis situations. Future research about HWFP should continue to explore multiple HWFP strategies, their barriers, and their potentially myriad relationships to food security, diet, and health outcomes, especially with longitudinal data., (© 2024. The Author(s).)

Acute kidney injury (AKI) is a frequent complication following allogeneic hematopoietic stem cell transplantation (allo-HSCT), but few studies have focused on AKI treated with kidney replacement therapy (AKI-KRT), particularly among critically ill patients. We investigated the incidence, risk factors, and 90-day mortality associated with AKI-KRT in 529 critically ill adult allo-HSCT recipients admitted to the ICU within 1-year post-transplant at two academic medical centers between 2011 and 2021. AKI-KRT occurred in 111 of the 529 patients (21.0%). Lower baseline eGFR, veno-occlusive disease, thrombotic microangiopathy, admission to an ICU within 90 days post-transplant, and receipt of invasive mechanical ventilation (IMV), total bilirubin ≥5.0 mg/dl, and arterial pH <7.40 on ICU admission were each associated with a higher risk of AKI-KRT. Of the 111 patients with AKI-KRT, 97 (87.4%) died within 90 days. Ninety-day mortality was 100% in each of the following subgroups: serum albumin ≤2.0 g/dl, total bilirubin ≥7.0 mg/dl, arterial pH ≤7.20, IMV with moderate-to-severe hypoxemia, and ≥3 vasopressors/inotropes at KRT initiation. AKI-KRT was associated with a 6.59-fold higher adjusted 90-day mortality in critically ill allo-HSCT vs. non-transplanted patients. Short-term mortality remains exceptionally high among critically ill allo-HSCT patients with AKI-KRT, highlighting the importance of multidisciplinary discussions prior to KRT initiation., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

Objective: Examine the relationship between weight trajectory and 2-year neurodevelopmental outcomes for extremely low birthweight (ELBW) infants with BPD., Study Design: Secondary analysis of infants born from 2010 to 2019. The predictor was BPD severity and the outcome was neurodevelopmental impairment, defined as any Bayley Scales of Infant Development (BSID) III score <70 at 24 months' corrected age. Repeated measures logistic regression was performed., Results: In total, 5042 infants were included. Faster weight trajectory was significantly associated with a decreased probability of having at least one BSID III score <70 for infants with grade 1-2 BPD (p < 0.0001) and an increased probability of at least one BSID III score <70 for infants with grade 3 BPD (p < 0.009). There was no significant association between weight trajectory and BSID III score <70 for infants with grade 0 BPD., Conclusion: The association between postnatal weight trajectory and neurodevelopmental outcome in this study differs by BPD severity., (© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)

LRRK2 is one of the most promising drug targets for Parkinson's disease. Though type I kinase inhibitors of LRRK2 are under clinical trials, alternative strategies like type II inhibitors are being actively pursued due to the potential undesired effects of type I inhibitors. Currently, a robust method for LRRK2-inhibitor structure determination to guide structure-based drug discovery is lacking, and inhibition mechanisms of available compounds are also unclear. Here we present near-atomic-resolution structures of LRRK2 with type I (LRRK2-IN-1 and GNE-7915) and type II (rebastinib, ponatinib, and GZD-824) inhibitors, uncovering the structural basis of LRRK2 inhibition and conformational plasticity of the kinase domain with molecular dynamics (MD) simulations. Type I and II inhibitors bind to LRRK2 in active-like and inactive conformations, so LRRK2-inhibitor complexes further reveal general structural features associated with LRRK2 activation. Our study provides atomic details of LRRK2-inhibitor interactions and a framework for understanding LRRK2 activation and for rational drug design., (© 2024. The Author(s).)

In eukaryotic cells, ATP generation is generally viewed as the primary function of mitochondria under normoxic conditions. Reactive oxygen species (ROS), in contrast, are regarded as the by-products of respiration, and are widely associated with dysfunction and disease. Important signaling functions have been demonstrated for mitochondrial ROS in recent years. Still, their chemical reactivity and capacity to elicit oxidative damage have reinforced the idea that ROS are the products of dysfunctional mitochondria that accumulate during disease. Several studies support a different model, however, by showing that: (1) limited oxygen availability results in mitochondria prioritizing ROS production over ATP, (2) ROS is an essential adaptive mitochondrial signal triggered by various important stressors, and (3) while mitochondria-independent ATP production can be easily engaged by most cells, there is no known replacement for ROS-driven redox signaling. Based on these observations and other evidence reviewed here, we highlight the role of ROS production as a major mitochondrial function involved in cellular adaptation and stress resistance. As such, we propose a rekindled view of ROS production as a primary mitochondrial function as essential to life as ATP production itself., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

Objective: To quantify the association between necrotizing enterocolitis (NEC) and neurodevelopmental disability (NDI) in extremely low birth weight (ELBW) infants with intraventricular hemorrhage (IVH)., Study Design: ELBW survivors born 2011-2017 and evaluated at 16-26 months corrected age in the Vermont Oxford Network (VON) ELBW Follow-Up Project were included. Logistic regression determined the adjusted relative risk (aRR) of severe NDI in medical or surgical NEC compared to no NEC, stratified by severity of IVH., Results: Follow-up evaluation occurred in 5870 ELBW survivors. Compared to no NEC, medical NEC had no impact on NDI, regardless of IVH status. Surgical NEC increased risk of NDI in patients with no IVH (aRR 1.69; 95% CI 1.36-2.09), mild IVH (aRR 1.36;0.97-1.92), and severe IVH (aRR 1.35;1.13-1.60)., Conclusions: ELBW infants with surgical NEC carry increased risk of neurodevelopmental disability within each IVH severity stratum. These data describe the additive insult of surgical NEC and IVH on neurodevelopment, informing prognostic discussions and highlighting the need for preventative interventions., (© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Serine Threonine Kinase 11 (STK11) loss of function (LoF) correlates with anti-PD-1 therapy resistance in patients with KRAS-driven lung adenocarcinoma (LUAD). The molecular mechanisms governing this observation remain unclear and represent a critical outstanding question in the field of lung oncology. As an initial approach to understand this phenomenon, we knocked-out (KO) STK11 in multiple KRAS-driven, STK11-competent human LUAD cell lines and performed whole transcriptome analyses to identify STK11-loss-dependent differential gene expression. Subsequent pathway enrichment studies highlighted activation of the HIPPO/YAP1 signaling axis, along with the induction of numerous tumor-intrinsic cytokines. To validate that YAP1-mediated transcriptional activation occurs in response to STK11 loss, we pursued YAP1 perturbation as a strategy to restore an STK11-competent gene expression profile in STK11-KO LUAD cell lines. Together, our data link STK11 loss with YAP1-mediated transcriptional activation, including the upregulation of immune-evasion promoting cytokines IL-6, CXCL8 and CXCL2. Further, our results raise the intriguing possibility that YAP1 antagonism may represent a therapeutic approach to counter anti-PD-1 therapy resistance in STK11-null, KRAS-driven LUADs by modulating tumor-intrinsic gene expression to promote a "hot" tumor immune microenvironment., (© 2023. The Author(s).)

Background: To characterize neurodevelopmental abnormalities in children up to 36 months of age with congenital Zika virus exposure., Methods: From the U.S. Zika Pregnancy and Infant Registry, a national surveillance system to monitor pregnancies with laboratory evidence of Zika virus infection, pregnancy outcomes and presence of Zika associated birth defects (ZBD) were reported among infants with available information. Neurologic sequelae and developmental delay were reported among children with ≥1 follow-up exam after 14 days of age or with ≥1 visit with development reported, respectively., Results: Among 2248 infants, 10.1% were born preterm, and 10.5% were small-for-gestational age. Overall, 122 (5.4%) had any ZBD; 91.8% of infants had brain abnormalities or microcephaly, 23.0% had eye abnormalities, and 14.8% had both. Of 1881 children ≥1 follow-up exam reported, neurologic sequelae were more common among children with ZBD (44.6%) vs. without ZBD (1.5%). Of children with ≥1 visit with development reported, 46.8% (51/109) of children with ZBD and 7.4% (129/1739) of children without ZBD had confirmed or possible developmental delay., Conclusion: Understanding the prevalence of developmental delays and healthcare needs of children with congenital Zika virus exposure can inform health systems and planning to ensure services are available for affected families., Impact: We characterize pregnancy and infant outcomes and describe neurodevelopmental abnormalities up to 36 months of age by presence of Zika associated birth defects (ZBD). Neurologic sequelae and developmental delays were common among children with ZBD. Children with ZBD had increased frequency of neurologic sequelae and developmental delay compared to children without ZBD. Longitudinal follow-up of infants with Zika virus exposure in utero is important to characterize neurodevelopmental delay not apparent in early infancy, but logistically challenging in surveillance models., (© 2023. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Climate projections predict major changes in alpine environments by the end of the 21st century. To avoid climate-induced maladaptation and extinction, many animal populations will either need to move to more suitable habitats or adapt in situ to novel conditions. Since populations of a species exhibit genetic variation related to local adaptation, it is important to incorporate this variation into predictive models to help assess the ability of the species to survive climate change. Here, we evaluate how the adaptive genetic variation of a mountain ungulate-the Northern chamois (Rupicapra rupicapra)-could be impacted by future global warming. Based on genotype-environment association analyses of 429 chamois using a ddRAD sequencing approach, we identified genetic variation associated with climatic gradients across the European Alps. We then delineated adaptive genetic units and projected the optimal distribution of these adaptive groups in the future. Our results suggest the presence of local adaptation to climate in Northern chamois with similar genetic adaptive responses in geographically distant but climatically similar populations. Furthermore, our results predict that future climatic changes will modify the Northern chamois adaptive landscape considerably, with various degrees of maladaptation risk., (© 2023. The Author(s), under exclusive licence to The Genetics Society.)

The network reconstruction task aims to estimate a complex system's structure from various data sources such as time series, snapshots, or interaction counts. Recent work has examined this problem in networks whose relationships involve precisely two entities-the pairwise case. Here, using Bayesian inference, we investigate the general problem of reconstructing a network in which higher-order interactions are also present. We study a minimal example of this problem, focusing on the case of hypergraphs with interactions between pairs and triplets of vertices, measured imperfectly and indirectly. We derive a Metropolis-Hastings-within-Gibbs algorithm for this model to highlight the unique challenges that come with estimating higher-order models. We show that this approach tends to reconstruct empirical and synthetic networks more accurately than an equivalent graph model without higher-order interactions., (© 2023. The Author(s).)

Objective: Describe the frequency of best practice behaviors during NICU provider and nursing shift-to-shift handoffs and identify strengths and opportunities for improvement., Study Design: Observational study of handoff characteristics among 40 centers participating in a learning collaborative over a 10-month period. Data were gathered using a handoff audit tool that outlined best practices. Comparisons of behaviors between nurse-to-nurse and provider-to-provider handoffs were made where appropriate., Results: Overall, 946 audits of shift-to-shift handoffs were analyzed. While many behaviors were demonstrated reliably, differences between nurse-to-nurse vs provider-to-provider handoffs were noted. Families were present for 5.9% of handoffs and, among those who were present, 48.2% participated by contributing information, asking questions, and sharing goals., Conclusions: Observation and measurement of handoff behaviors can be used to identify opportunities to improve handoff communication, family participation, and human factors that support handoff. Auditing handoffs is feasible and necessary to improve these critical transitions in infants' care., (© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Background: Maternal stress has negative consequences on infant behavioral development, and COVID-19 presented uniquely stressful situations to mothers of infants born during the pandemic. We hypothesized that mothers with higher levels of perceived stress during the pandemic would report higher levels of infant regulatory problems including crying and interrupted sleep patterns., Methods: As part 6 sites of a longitudinal study, mothers of infants born during the pandemic completed the Perceived Stress Scale, the Brief Infant Sleep Questionnaire, and an Infant Crying survey at 6 (n = 433) and 12 (n = 344) months of infant age., Results: Maternal perceived stress, which remained consistent at 6 and 12 months of infant age, was significantly positively correlated with time taken to settle infants. Although maternal perceived stress was not correlated with uninterrupted sleep length, time taken to put the infant to sleep was correlated. Perceived stress was also correlated with the amount of infant crying and fussiness reported at 6 months., Conclusions: Mothers who reported higher levels of perceived stress during the pandemic reported higher levels of regulatory problems, specifically at 6 months. Examining how varying levels of maternal stress and infant behaviors relate to overall infant developmental status over time is an important next step., Impact: Women giving birth during the COVID-19 pandemic who reported higher levels of stress on the Perceived Stress Scale also reported higher levels of infant fussiness and crying at 6 months old, and more disruptive sleep patterns in their infants at 6 months and 12 months old. Sleeping problems and excessive crying in infancy are two regulatory problems that are known risk factors for emotional and behavioral issues in later childhood. This paper is one of the first studies highlighting the associations between maternal stress and infant behaviors during the COVID-19 pandemic., (© 2023. The Author(s).)

To optimize post-graduate competency-based assessment for medical trainees, the Accreditation Council for Graduate Medical Education initiated a sub-specialty-specific revision of the existing Milestones 1.0 assessment framework in 2016. This effort was intended to increase both the effectiveness and accessibility of the assessment tools by incorporating specialty-specific performance expectations for medical knowledge and patient care competencies; decreasing item length and complexity; minimizing inconsistencies across specialties through the development of common "harmonized" milestones; and providing supplemental materials, including examples of expected behaviors at each developmental level, suggested assessment strategies, and relevant resources. This manuscript describes the efforts of the Neonatal-Perinatal Medicine Milestones 2.0 Working Group, outlines the overall intent of Milestones 2.0, compares the novel Milestones to the original version, and details the materials contained in the novel supplemental guide. This new tool should enhance NPM fellow assessment and professional development while maintaining consistent performance expectations across specialties., (© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Higher rates of cardiovascular events have been observed among rural residents compared with urban. Hypertension and lack of blood pressure (BP) control are risk factors for cardiovascular events. We compared the prevalence of hypertension and controlled BP, and the distribution of systolic blood pressure (SBP), by urban-rural residence. Participants from the REasons for Geographic And Racial Differences in Stroke (REGARDS) Study, a prospective cohort of Black and White adults aged ≥45 years, were categorized as either urban, large rural, or small-isolated rural, by using the Rural-Urban Commuting Area (RUCA) categorization B system. Oucomes were hypertension prevalence (BP ≥ 140/90 mmHg or antihypertensive use), BP control (BP < 140/90 among participants on antihypertensive medication), and the distribution of SBP. Counfounders were age, race, sex, antihypertensive medication use, and US Census Bureau division. The analysis included 26,133 participants (80.3% urban, 11.6% large-rural, 8.2% small-isolated rural). The unadjusted prevalence of hypertension was not different between groups. However, after adjustment, the odds of hypertension was higher among participants in the large rural group (odds ratio [OR] 1.17; 95% confidence interval [CI], 1.08-1.27) and small-isolated rural group (OR 1.19; 95% CI, 1.08-1.30), compared with the urban group. There was no evidence of an adjusted difference in BP control for those taking antihypertensive medications. Adjusted differences in SBP were greater for both rural groups, compared with urban, at the higher percentiles of SBP. Rural residence was associated with a higher adjusted odds of hypertension and higher SBP., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

The skeleton forms from multipotent human mesenchymal stem cells (hMSCs) competent to commit to specific lineages. Long noncoding RNAs (lncRNAs) have been identified as key epigenetic regulators of tissue development. However, regulation of osteogenesis by lncRNAs as mediators of commitment to the bone phenotype is largely unexplored. We focused on LINC01638, which is highly expressed in hMSCs and has been studied in cancers, but not in regulating osteogenesis. We demonstrated that LINC01638 promotes initiation of the osteoblast phenotype. Our findings reveal that LINC01638 is present at low levels during the induction of osteoblast differentiation. CRISPRi knockdown of LINC01638 in MSCs prevents osteogenesis and alkaline phosphatase expression, inhibiting osteoblast differentiation. This resulted in decreased MSC growth rate, accompanied by double-strand breaks, DNA damage, and cell senescence. Transcriptome profiling of control and LINC01638-depleted hMSCs identified > 2000 differentially expressed mRNAs related to cell cycle, cell division, spindle formation, DNA repair, and osteogenesis. Using ChIRP-qPCR, molecular mechanisms of chromatin interactions revealed the LINC01638 locus (Chr 22) includes many lncRNAs and bone-related genes. These novel findings identify the obligatory role for LINC01638 to sustain MSC pluripotency regulating osteoblast commitment and growth, as well as for physiological remodeling of bone tissue., (© 2023. The Author(s).)

Emerging research indicates that high HDL-C levels might not be cardioprotective, potentially worsening cardiovascular disease (CVD) outcomes. Yet, there is no data on HDL-C's association with other CVD risk factors like myocardial fibrosis, a key aspect of cardiac remodeling predicting negative outcomes. We therefore aimed to study the association between HDL-C levels with interstitial myocardial fibrosis (IMF) and myocardial scar measured by CMR T1-mapping and late-gadolinium enhancement (LGE), respectively. There were 1863 participants (mean age of 69 years) who had both serum HDL-C measurements and underwent CMR. Analysis was done among those with available indices of interstitial fibrosis (extracellular volume fraction [ECV]; N = 1172 and native-T1; N = 1863) and replacement fibrosis by LGE (N = 1172). HDL-C was analyzed as both logarithmically-transformed and categorized into < 40 (low),40-59 (normal), and ≥ 60mg/dL (high). Multivariable linear and logistic regression models were constructed to assess the associations of HDL-C with CMR-obtained measures of IMF, ECV% and native-T1 time, and myocardial scar, respectively. In the fully adjusted model, each 1-SD increment of log HDL-C was associated with a 1% increment in ECV% (p = 0.01) and an 18-ms increment in native-T1 (p < 0.001). When stratified by HDL-C categories, those with high HDL-C (≥ 60mg/dL) had significantly higher ECV (β = 0.5%, p = 0.01) and native-T1 (β = 7 ms, p = 0.01) compared with those with normal HDL-C levels. Those with low HDL-C were not associated with IMF. Results remained unchanged after excluding individuals with a history of myocardial infarction. Neither increasing levels of HDL-C nor any HDL-C category was associated with the prevalence of myocardial scar. Increasing levels of HDL-C were associated with increased markers of IMF, with those with high levels of HDL-C being linked to subclinical fibrosis in a community-based setting., (© 2023. The Author(s).)

The combination of mirror therapy (MT) and neuromuscular electrical stimulation (NMES) has been devised as an intervention method in stroke rehabilitation; however, few studies have investigated its efficacy in lower extremity motor function recovery. In this systematic review and meta-analysis, we examined the effectiveness of combined MT and NMES therapy in improving poststroke walking speed, spasticity, balance and other gait parameters. Randomized controlled trials (RCTs) were selected from PubMed, Cochrane Library, EMBASE, and Scopus databases. In total, six RCTs which involving 181 participants were included. Our findings indicate that MT combined with NMES elicits greater improvement relative to control group in walking speed (SMD = 0.67, 95% confidence interval [CI] 0.26-1.07, P = 0.001), Berg Balance Scale (SMD = 0.72; 95% CI 0.31-1.13; P = 0.0007), cadence (SMD = 0.59, 95% CI 0.02-1.16, P = 0.04), step length (SMD = 0.94, 95% CI 0.35-1.53, P = 0.002), and stride length (SMD = 0.95, 95% CI 0.36-1.54, P = 0.002) but not in modified Ashworth scale (SMD =  - 0.40, 95% CI - 1.05 to 0.26, P = 0.23). Our findings suggest that MT combined with NMES may be a suitable supplemental intervention to conventional therapy in stroke survivors., (© 2023. The Author(s).)

Timely repair of chromosomal double-strand breaks is required for genome integrity and cellular viability. The polymerase theta-mediated end joining pathway has an important role in resolving these breaks and is essential in cancers defective in other DNA repair pathways, thus making it an emerging therapeutic target 1 . It requires annealing of 2-6 nucleotides of complementary sequence, microhomologies, that are adjacent to the broken ends, followed by initiation of end-bridging DNA synthesis by polymerase θ. However, the other pathway steps remain inadequately defined, and the enzymes required for them are unknown. Here we demonstrate requirements for exonucleolytic digestion of unpaired 3' tails before polymerase θ can initiate synthesis, then a switch to a more accurate, processive and strand-displacing polymerase to complete repair. We show the replicative polymerase, polymerase δ, is required for both steps; its 3' to 5' exonuclease activity for flap trimming, then its polymerase activity for extension and completion of repair. The enzymatic steps that are essential and specific to this pathway are mediated by two separate, sequential engagements of the two polymerases. The requisite coupling of these steps together is likely to be facilitated by physical association of the two polymerases. This pairing of polymerase δ with a polymerase capable of end-bridging synthesis, polymerase θ, may help to explain why the normally high-fidelity polymerase δ participates in genome destabilizing processes such as mitotic DNA synthesis 2 and microhomology-mediated break-induced replication 3 ., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

Nano-scale extracellular vesicles are lipid-bilayer delimited particles that are naturally secreted by all cells and have emerged as valuable biomarkers for a wide range of diseases. Efficient isolation of small extracellular vesicles while maintaining yield and purity is crucial to harvest their potential in diagnostic, prognostic, and therapeutic applications. Most conventional methods of isolation suffer from significant shortcomings, including low purity or yield, long duration, need for large sample volumes, specialized equipment, trained personnel, and high costs. To address some of these challenges, our group has reported a novel insulator-based dielectrophoretic device for rapid isolation of small extracellular vesicles from biofluids and cell culture media based on their size and dielectric properties. In this study, we report a comprehensive characterization of small extracellular vesicles isolated from cancer-patients' biofluids at a twofold enrichment using the device. The three-fold characterization that was performed using conventional flow cytometry, advanced imaging flow cytometry, and microRNA sequencing indicated high yield and purity of the isolated small extracellular vesicles. The device thus offers an efficient platform for rapid isolation while maintaining biomolecular integrity., (© 2023. Springer Nature Limited.)

Despite the prognostic value of arterial stiffness (AS) and pulsatile hemodynamics (PH) for cardiovascular morbidity and mortality, epigenetic modifications that contribute to AS/PH remain unknown. To gain a better understanding of the link between epigenetics (DNA methylation) and AS/PH, we examined the relationship of eight measures of AS/PH with CpG sites and co-methylated regions using multi-ancestry participants from Trans-Omics for Precision Medicine (TOPMed) Multi-Ethnic Study of Atherosclerosis (MESA) with sample sizes ranging from 438 to 874. Epigenome-wide association analysis identified one genome-wide significant CpG (cg20711926-CYP1B1) associated with aortic augmentation index (AIx). Follow-up analyses, including gene set enrichment analysis, expression quantitative trait methylation analysis, and functional enrichment analysis on differentially methylated positions and regions, further prioritized three CpGs and their annotated genes (cg23800023-ETS1, cg08426368-TGFB3, and cg17350632-HLA-DPB1) for AIx. Among these, ETS1 and TGFB3 have been previously prioritized as candidate genes. Furthermore, both ETS1 and HLA-DPB1 have significant tissue correlations between Whole Blood and Aorta in GTEx, which suggests ETS1 and HLA-DPB1 could be potential biomarkers in understanding pathophysiology of AS/PH. Overall, our findings support the possible role of epigenetic regulation via DNA methylation of specific genes associated with AIx as well as identifying potential targets for regulation of AS/PH., (© 2023. Springer Nature Limited.)

Previous studies have documented natural infections of SARS-CoV-2 in various domestic and wild animals. More recently, studies have been published noting the susceptibility of members of the Cervidae family, and infections in both wild and captive cervid populations. In this study, we investigated the presence of SARS-CoV-2 in mammalian wildlife within the state of Vermont. 739 nasal or throat samples were collected from wildlife throughout the state during the 2021 and 2022 harvest season. Data was collected from red and gray foxes (Vulpes vulples and Urocyon cineroargentus, respectively), fishers (Martes pennati), river otters (Lutra canadensis), coyotes (Canis lantrans), bobcats (Lynx rufus rufus), black bears (Ursus americanus), and white-tailed deer (Odocoileus virginianus). Samples were tested for the presence of SARS-CoV-2 via quantitative RT-qPCR using the CDC N1/N2 primer set and/or the WHO-E gene primer set. Surprisingly, we initially detected a number of N1 and/or N2 positive samples with high cycle threshold values, though after conducting environmental swabbing of the laboratory and verifying with a second independent primer set (WHO-E) and PCR without reverse transcriptase, we showed that these were false positives due to plasmid contamination from a construct expressing the N gene in the general laboratory environment. Our final results indicate that no sampled wildlife were positive for SARS-CoV-2 RNA, and highlight the importance of physically separate locations for the processing of samples for surveillance and experiments that require the use of plasmid DNA containing the target RNA sequence. These negative findings are surprising, given that most published North America studies have found SARS-CoV-2 within their deer populations. The absence of SARS-CoV-2 RNA in populations sampled here may provide insights in to the various environmental and anthropogenic factors that reduce spillover and spread in North American's wildlife populations., (© 2023. Springer Nature Limited.)

Lentiviral vectors are attractive delivery vehicles for cystic fibrosis gene therapy owing to their low immunogenicity and ability to integrate into the host cell genome, thereby producing long-term, stable gene expression. Nonetheless, repeat dosing may be required to increase initial expression levels, and/or boost levels when they wane. The primary aim of this study was to determine if repeat dosing of a VSV-G pseudotyped LV vector delivered into mouse lungs is more effective than a single dose. C57Bl/6 mouse lungs were conditioned with lysophosphatidylcholine, followed one-hour later by a LV vector carrying the luciferase reporter gene, using six different short-term (≤1 wk) and long-term (>1 wk) dosing schedules. Luciferase expression was quantified using bioluminescence imaging over 12 months. Most dosing schedules produced detectable bioluminescence over the 12-month period, but the shorter intervals (≤1 wk) produced higher levels of flux than the longest interval (five doses at least 1-month apart). Ex vivo lung analysis at 12 months showed that the estimated mean flux for the group that received two doses 1-week apart was significantly greater than the single dose group and the two groups that received doses over a period greater than 1-week. These results suggest that early consecutive multiple doses are more effective at improving gene expression in mouse lungs at 12 months, than longer repeat dosing intervals., (© 2023. The Author(s).)