Open Research Europe

BackgroundMalaria control is increasingly challenged by the urban-adapted vector Anopheles stephensi, yet molecular and cellular tools for this species remain scarce, restricting functional genomic studies and the development of genetic control strategies. To help address this gap, we established a new embryo-derived Anopheles stephensi cell line. ResultsWe generated and characterised a novel embryo-derived Anopheles stephensi (SDA-500) cell line capable of sustained growth in vitro. Species identity was confirmed by mitochondrial COI barcoding, and karyotypic analysis revealed a diploid chromosome complement with the presence of a Y chromosome, confirming that at least some cells are of male origin. Transfection conditions were optimized, with TransIT-PRO showing higher efficiency than Lipofectamine-based reagents. Using a dual-luciferase reporter assay, of several promoters tested the Anopheles gambiae polyubiquitin promoter exhibited the strongest and most consistent transcriptional activity in SDA-500 cells. ConclusionsThe SDA-500 cell line provides a stable and genetically validated in vitro platform that supports efficient transgene expression. This resource provides a useful system for functional genomics and molecular manipulation in Anopheles stephensi and is expected to facilitate studies of mosquito biology and contribute to the development of novel malaria control strategies.

Occupational ocular injuries are a preventable yet neglected public health problem, particularly in low- and middle-income countries. Maintenance workers are exposed to diverse ocular hazards daily, yet compliance with protective measures is consistently poor. A descriptive cross-sectional study was conducted among 85 maintenance workers at the Maintenance and Essential Services Organization (MESO) of Kwame Nkrumah University of Science and Technology (KNUST), Ghana, recruited through stratified convenience sampling across seven occupational sections. A structured questionnaire assessed knowledge of ocular hazards and protective equipment, attitudes toward ocular safety, and safety practices. Data were analyzed using IBM SPSS version 26 (IBM Corp., Armonk, NY, USA); chi-square and Fishers exact tests assessed associations (p < 0.05). Participants were predominantly male (84/85, 98.8%), with a mean age of 44.5 {+/-} 10.4 years. Overall knowledge was good (mean 9.40 {+/-} 1.59 out of 11), but attitude and practice scores were average (2.78 {+/-} 0.92 and 3.27 {+/-} 0.93, respectively). Most workers correctly identified goggles and face shields as protective, but only about half recognized that ordinary sunglasses and spectacles offer inadequate protection. Although 97.6% (83/85) recognized the need for ocular protection, only 7.1% (6/85) reported consistent protective eyewear use, and fewer than half (45.9%, 39/85) had received formal ocular safety training. Routine general protective equipment use was significantly associated with ocular protection use (Fishers exact test, p = 0.011). Sand and dust particles were the leading causes of injury and only 25% (5/20) of injured workers sought formal care. Workers demonstrated good knowledge but poor attitudes and practices toward ocular safety, suggesting that knowledge alone does not translate into protective behaviour even within a relatively well-resourced institutional setting. Findings suggest that limited access to task-appropriate protective eyewear may represent an important institutional barrier. Institutional PPE supply and section-specific safety training are essential to bridge this knowledge-practice gap.

Uniform, reproducible blood smears are critical for reliable hematological evaluation. Manual smear preparation, however, is user-dependent and introduces variability that limits quantitative microscopy. Here we developed BUsmear, a low-cost, 3D-printed, motorized blood smear device that prepares two smears simultaneously from a printed stage and a micro-motor drive with tunable linear velocity, controlled through a joystick-operated driver module. By spreading two slides in parallel with fully repeatable slide-to-slide motion, the device doubles throughput while eliminating operator-dependent motion artifacts, and can be fabricated on any benchtop 3D printer in under one day. To validate smear quality, we analyzed blood films from three donors together with a manual smear prepared by an expert from the blood of one of the same donors, giving a matched device-versus-manual pair. Automated Cellpose segmentation of 14,046 red blood cells across 12 bright-field fields showed that cell diameter was preserved and closely matched the expert smear (6.7-7.8 um across groups, within the 6.2-8.2 um human reference range; 6.7 vs 6.9 um in the matched pair), and that all films formed non-aggregated monolayers (Clark-Evans index of aggregation 1.04-1.24). Critically, red blood cells in the device films were markedly more circular than in the expert manual smear (mean eccentricity 0.405 vs 0.502; 0.443 vs 0.502 in the matched pair), with complete separation between the two methods at the level of whole fields of view. Because eccentricity reports smear-induced cell distortion, this indicates that a constant, mechanically controlled spreading velocity preserves red blood cell morphology better than skilled manual technique. BUsmear offers an accessible route to standardized smear geometry for quantitative analysis, including AI-based morphometry, and its low cost may be particularly advantageous in low-income countries with a high prevalence of malaria.

The photic sneeze reflex (PSR), in which exposure to bright light triggers sneezing, is widely recognised but inconsistently defined and measured. We conducted the first systematic review and meta-analysis of PSR prevalence to synthesise the epidemiological evidence, assess methodological quality, and identify priorities for future research. We included 18 articles comprising 31 study groups and extracted prevalence estimates, study characteristics, ascertainment methods, and epidemiological information. Fifteen eligible study groups classified as healthy were included in the primary meta-analysis. The pooled prevalence was 22% (95% CI, 15%-29%), with extreme between-study heterogeneity (I2 = 99.3%). Reported prevalence estimates and associations with participant characteristics varied widely, and nearly all studies were judged to be at high risk of bias. The pooled estimate should therefore be interpreted as a descriptive summary of the available evidence rather than a precise estimate of population prevalence. Future studies require a standardised operational definition, representative sampling, transparent reporting, and reproducible methods for assessing light-triggered sneezing.

Previously, we have demonstrated that certain ferric salts common in Archean waters, such as iron trichloride and ferric ammonium citrate, can protect liposomes from destruction by short-wavelength UVC light. In this study, we investigate the propagation of 254 nm UV radiation through aqueous FeCl3 solutions and its interactions with liposomes. We then consider these findings in the context of early Earth UV environment, discuss their implications for our hypothesis of the Darwinian evolution of liposomes, and integrate them with our previous experimental results.

Background In Nigeria, the country with the greatest global malaria burden, funding constraints increasingly require insecticide-treated net (ITN) reprioritization to target those at highest risk. Large language models (LLM)- assisted decision-support tools may facilitate risk-informed ITN planning by supporting malaria programme officers in navigating analyses, interpreting outputs, and translating evidence into operational decisions. We developed ChatMRPT, an LLM-assisted ITN allocation planning tool based on user requirements, and analyzed post-interaction feedback, examined user digital competencies and workplace resources, and identified institutionalization pathways for LLM-assisted intervention planning. Methods A two-phase mixed-methods study began with software requirements gathering workshops (using a prototype) involving representatives from the National Malaria Elimination Programme (NMEP), State Malaria Elimination Programmes (SMEPs), and implementing partners. Phase 2two evaluated ChatMRPT through surveys, guided exercises, and focus group discussions with 34 SMEP officers from 28 Nigerian states. Quantitative data were analyzed using descriptive statistics and profile-based comparisons, while qualitative data were analyzed using reflexive thematic analysis to synthesize user experiences of ChatMRPT and identify institutionalization pathways. Findings Fifty-eight percent (19/33) of participants demonstrated both higher digital competency and adequate workplace resources; the remainder exhibited limitations in one or both domains ([4/33] higher competency/constrained resources; [6/33] higher resources/lower competency). Participants with higher digital competency but constrained workplace resources reported user experiences comparable to those with higher competency and adequate resources, whereas workplace resources alone did not appear to compensate for lower digital competency. Key software requirements included contextual guidance for malaria risk interpretation, operational decision support, and embedded analytical support. Following iterative incorporation of these requirements, ChatMRPT was positively evaluated across participant profiles. Participants viewed institutionalization as dependent on integration into routine malaria planning and adaptability to evolving programme priorities. Interpretation Many malaria programme officers may already have the foundational competency for LLM-assisted decision support. However, there is room to further strengthen digital competencies while facilitating access to basic workplace resources such as stable internet. Institutionalization of LLM tools may depend on addressing these capacity and infrastructural constraints alongside designing explainable, integrated, and flexible systems. Future research should evaluate long-term integration, sustainability, and effectiveness in routine malaria planning. Funding This work was funded by the Bill and Melinda Gates Foundation (INV-036449) and the Center for Health Outcomes and Informatics Research (CHOIR), Loyola University Chicago. The funders had no role in the study design, data analysis, interpretation of findings, or preparation of the manuscript.

Background Scientific mistrust contributes to lower participation and underrepresentation of Black Americans in genomics studies limiting understanding of how genomic variation and environmental exposures influence health disparities. Community-engaged research requires rebuilding scientific trust; however, there is a need for practical models that operationalize guidance for researchers without community-engaged research training. Therefore, we developed an educational intervention for genomics researchers initiating partnerships with Black American communities. Our intervention creates a bidirectional teaching environment that allows potential community and academic partners to discuss areas of expertise for each stakeholder, partnership perspectives and needs, while exploring modules related to genomics research and community-academic partnership. We report a novel and structured approach for prospective academic and community partners to mutually orient one another and assess partnership practicability. Methods Ten participants, recruited through established community channels, attended a four-hour workshop containing nine interactive modules about scientific mistrust, research safeguards, community-engaged research, genomics, and research for community-defined goals. We also experimented with humor to enhance engagement and trust. We used mixed-methods, single-arm research design and assessed feasibility through recruitment success and retention. Using inductive rapid thematic analyses, we assessed participant responses to eight workshop prompts. Preliminary quantitative data, used for descriptive purposes due to low sample size, were analyzed from pre- and post-Likert scale surveys assessing associations between the intervention and four domains, including scientific trust. Results All 10 enrolled participants completed the study, meeting a priori criteria of 100% for recruitment and show rate. Engagement was strongest during early workshop modules and declined in later modules. Survey data completeness was limited by missing responses. Survey instrument design limitations were identified for modification in future studies. Participants articulated expectations for partnership that aligned with community-based participatory research principles. Conclusions The intervention is feasible to deliver as a bidirectional educational experience in partnership with a community organization. The strongest implementation refinements needed were workshop duration, module prioritization, and survey instrument design, particularly the trust domain. A community advisory board is co-developing modules and refining the intervention to evaluate in a pilot version of the study with a larger sample.

Quantification of microbial growth inhibition is central to assays ranging from antibiotic susceptibility of bacteria to sensitivity of yeasts to antifungal therapeutics. Classical analysis approaches derive from zone-of-inhibition (termed halo) formats using filter paper discs, spanning methods from laser detection to machine learning. However, these tools struggle with non-uniform halos, fail to account for lawn density variability despite its experimental influence, and lack accessible, reproducible code. Here, we present Halo Unbiased Measurement of growth Inhibition (HaloUMI); an open-source Python graphical user interface for automated, high-throughput analysis of lawn-based microbial assays. HaloUMI integrates robust image processing with physics-informed models to quantify inhibition zones irrespective of shape, enabling accurate segmentation of uniform and irregular halo phenotypes. This analysis pipeline incorporates the critical correction for spatial heterogeneity in lawn density, improving reproducibility across experimental conditions. The software enhances usability without sacrificing precision, allowing rapid batch processing and intuitive parameter control. HaloUMI can be applied to multiple assay types, including yeast toxin halo, microbial mating, and conventional filter paper disc assays. It yields high-precision measurement of halo size and morphology, with improved consistency compared to standard thresholding and circular fitting. By combining accessibility, flexibility, and biophysical modelling, HaloUMI provides a quantitative framework for irregularly shaped halos of lawns of varying growth potential, enabling generalisable analysis of broad microbial interactions. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=84 SRC="FIGDIR/small/743694v1_ufig1.gif" ALT="Figure 1"> View larger version (20K): org.highwire.dtl.DTLVardef@1bd7c88org.highwire.dtl.DTLVardef@13ac9b6org.highwire.dtl.DTLVardef@9108e1org.highwire.dtl.DTLVardef@1de06bd_HPS_FORMAT_FIGEXP M_FIG C_FIG

Operant paradigms are powerful tools to quantify motivation and reward. Traditionally, operant conditioning research has been limited to food and drug reinforcers. Recent advances in commercially available operant equipment, however, allow for the quantification of social motivation. These operant assays are an improvement over commonly used social preference tasks, as they enable direct measurement of the effort and motivation driving social behavior. Based on a design by Venniro et al. (2020), the MedPC social operant boxes modify the traditional operant box setup for social interactions. The experimental rat can lever-press to raise a door for an interaction with a target rat behind a porous barrier. These social operant boxes have been widely adapted to test social behavior in adult and adolescent rodents and investigate how a range of conditions (e.g. stress, drug taking, etc.) affect social motivation. However, there is a gap in assessing maternal motivation for pups during the postpartum period, despite ample evidence that postpartum social behavior is highly relevant for offspring health outcomes. Here, we detail 3D-printed modifications to the standard Med PC social operant boxes to adapt the social target chamber to safely house neonatal pups. We have also developed testing protocols to assess motivation during the limited postpartum period. These data demonstrate that, with simple modifications to social operant chambers and testing protocols, the field can implement advanced behavioral approaches to directly assess maternal motivation.

The European leaf-toed gecko (Euleptes europaea) is a small, nocturnal gecko endemic to the western Mediterranean. As a phylogenetically distinctive member of the Gondwanan family Sphaerodactylidae, it represents an important species for studying Mediterranean island biogeography, adaptation, and reptile genome evolution. The species also occupies a key position for investigating the evolution of sex chromosomes, as geckos exhibit remarkable diversity and frequent transitions in sex-determination systems. We present a chromosome-level genome assembly of Euleptes europaea generated as part of the Vertebrate Genomes Project. The 1.8 Gb assembly has a scaffold N50 of 102.3 Mb (contig N50 27 Mb), with 21 chromosome-scale scaffolds corresponding to the known karyotype (2n = 42). The primary assembly has a BUSCO completeness of 97.80% (95.60% as single-copy), a k-mer completeness of 96.00%, and a k-mer quality value (QV) of 61.20. Repetitive elements account for 53.20% of the genome and genome annotation identified 18,633 protein-coding genes. This high-quality reference genome will facilitate studies of genome evolution, island adaptation, and sex chromosome evolution across geckos and other reptiles.

Prolonged low-temperature exposure may extend the preservation window of mammalian cells but can also disrupt cellular homeostasis and ultimately compromise cell viability. This study investigated the time-dependent phenotypic and transcriptomic responses of primary canine dermal fibroblasts to sustained hypothermic stress. Passage-three fibroblasts were continuously maintained at 15 for up to 15 days, with samples collected on Days 0, 3, 6, 9, 12, and 15. Cellular morphology, metabolic activity and viability, and apoptosis were evaluated using bright-field microscopy, Cell Counting Kit-8 assays, and Annexin V-FITC/propidium iodide flow cytometry, respectively. RNA sequencing was performed to characterize dynamic transcriptional changes throughout the exposure period. Early low-temperature exposure was associated with relatively preserved cellular morphology and viability, suggesting a transient adaptive response. With increasing exposure duration, fibroblasts exhibited progressive morphological deterioration, reduced metabolic activity, loss of adhesion, and increased apoptosis. Time-series transcriptomic analysis further revealed temporally coordinated and stage-dependent gene-expression programs associated with metabolic regulation, cellular stress responses, structural homeostasis, and cell survival. Integration of phenotypic and transcriptomic data demonstrated that the response of primary canine dermal fibroblasts to 15 was dynamic rather than linear, progressing from early adaptation to cumulative dysfunction during prolonged exposure. These findings provide a framework for defining the low-temperature tolerance of primary canine dermal fibroblasts and may inform the optimization of protocols for their short- to medium-term preservation and transportation.

Objective. Respiratory infections are the leading cause of illness at major sporting events, yet surveillance relies on athletes recognising and reporting symptoms. We evaluated whether continuous air sampling with point-of-care molecular testing could detect respiratory-virus nucleic acids in an elite team's congregate spaces during competition, and whether the resulting signals were operationally useful. Methods. We performed a prospective, descriptive environmental-surveillance study following the Canadian men's national soccer team across five host cities during the 2026 FIFA World Cup (3 June to 4 July 2026). InBio Apollo bioaerosol samplers ran continuously in up to four team-designated rooms per hotel (physiotherapy, meal, equipment, and coaches' room or hallway). Filters were changed approximately twice daily, eluted on-site, and tested with the Cepheid Xpert Xpress(R) SARS-CoV-2/Flu/RSV plus assay. A sample was considered positive if any cycle-threshold (Ct) value was reported, as less than 45, for a target. Results. Of 174 air filters, there were 13 detections of virus genetic material (9 SARS-CoV-2, 3 influenza A virus, 1 influenza B virus, 0 RSV). Detections were sparse early and clustered late in the tournament. An influenza A signal appeared the morning a player was sent home febrile, and SARS-CoV-2 signals coincided with visibly ill hotel staff, with signals falling after ill staff were excluded. Conclusion. Air sampling with point-of-care testing is feasible in the mobile environment of an elite team and can surface behavior-independent viral signals during competition that may offer opportunities for earlier precautionary actions.

Introduction: Veterinary Professionals are at risk of contracting zoonoses, including potential emerging infections. Personal Protective Equipment (PPE) could reduce infection transmission risk, but use of it in the profession is low. Understanding Veterinary Professionals experiences with PPE could help identify facilitators and barriers, therefore informing a future strategy to improve usage. Methods: Two focus group discussion with veterinary nurses and two with veterinarians took place at a tertiary small animal teaching hospital. With an interpretivist epistemology, transcriptions were inductively coded and analysed using thematic analysis. Results: Veterinary Professionals frequently reported underusing PPE, despite significant zoonotic risk. Friction in interdisciplinary relationships negatively impacted veterinary professionals experiences with PPE: differing views on risk, policy and PPE, contrasting perceptions of each other, and challenges with communication impacted PPE decision-making. Barriers included an absence of initial recognition of risk, a staff culture of prioritising patient health over risk to self, friction in response to others PPE use or lack thereof, an overly complex policy, and a lack of cultural and institutional reaction to occupationally contracted zoonoses. Facilitators included effective inter and intradisciplinary communication and previous personal experience with serious zoonoses. Conclusion: Veterinary Professionals experiences with PPE, are shaped by social and professional factors, such as interdisciplinary friction, perceptions of self and others, of risk and policy. Recommendations are for the findings to be used locally to embed PPE use into everyday practice. At a national level, the United Kingdom Health Security Agency (UKHSA) should use the findings to evaluate current policies regarding PPE and zoonoses in veterinary practice and produce a simplified national guidance, in collaboration with veterinary professionals with lived experience. Further research into General Practice Veterinary Professionals experiences and the interdisciplinary social dynamics is recommended.

Traditionally, complex behavioral tasks in mice have relied upon water restriction as an external motivator to increase task engagement. However, citric acid (CA) water, the technique whereby water is given ad libitum but made sour by the addition of CA, has emerged as an alternative to typical water restriction. Evidence suggests CA water can effectively motivate task performance while improving animal welfare in alignment with the 3Rs of animal research and reducing experimenter labor. While promising, the applicability of CA water in mice remains incompletely characterized with higher concentrations only tested in rats and "ramping" schedules, where mice progress to increasingly higher concentrations, indirectly examined. Here, we evaluate four CA concentration/schedule combinations for their effects on mouse health (weight changes, home cage behaviors, fecal counts) and motivation to drink regular water (lick counts, drinking behaviors) in female and male C57BL/6J mice. We find that a schedule ramping from 1% to 2% CA after one week is the easiest for mice to adapt to and sustained 2% CA use maintains robust lick counts for at least 5 weeks. Additionally, CA has been directly characterized for wheel-turning and touchscreen tasks, but not virtual reality (VR) tasks, an increasingly popular class of behavioral experiments. Therefore, we also assess how 2% CA affects motivation and task performance in two VR treadmill tasks (running and stopping task). We find that CA water does not improve task performance above that of mice given regular water, but does limit competing motivations and produce more uniform, reward-motivated behavior.

Behavioral thermoregulation allows animals to reduce the physiological costs associated with variation in the thermal environment, yet little is known about how chimpanzees respond behaviorally to the cool and humid conditions of tropical montane forests. We examined how microclimatic conditions influenced resting posture, sun/shade use, substrate use, vertical position within the canopy, and day nest use in eastern chimpanzees (Pan troglodytes schweinfurthii) in Nyungwe National Park, Rwanda. From March 2024 to February 2025, we collected 5,661 individual behavioral records during 486 contact hours and paired behavioral observations with measurements of ambient temperature, water vapor pressure, wind speed, and rainfall. Bayesian mixed models showed that temperature was the most consistent climatic correlate of resting behavior. Under cooler conditions, chimpanzees were more likely to adopt heat conserving postures, particularly curled and huddled postures, and to use day nests. As temperature increased, chimpanzees increased their use of shade, shifted from arboreal to terrestrial resting, and used lower canopy strata more frequently. Higher wind speed was also associated with reduced use of the upper canopy, whereas rainfall, water vapor pressure, and fruit availability generally showed weaker or behavior-specific associations. These findings indicate that chimpanzees in a cool montane forest flexibly adjust posture, solar exposure, substrate use, vertical position, and day nest use in relation to local thermal conditions. Resting behaviour may therefore provide an important context for behavioral thermoregulation, extending our understanding of chimpanzee thermal ecology beyond the warmer lowland and savanna habitats that have received most previous attention.

Background Madagascar has one of the highest burdens of schistosomiasis globally, yet epidemiological evidence from the South-east of the country remains limited. Existing surveillance estimates suggest relatively low prevalence in this region. This contrasts with the region's environmental suitability for transmission and its population's high occupational risk through rice farming. Methods A cross-sectional exploratory study was conducted across 13 villages around the Manombo Special Reserve, Farafangana District, between May and July 2025. Participants were surveyed for geographic, environmental and behavioural risk factors for schistosomiasis infection. Infection was diagnosed by Kato-Katz stool microscopy. Multivariable mixed-effects logistic and linear regression, with village as a random intercept, estimated associations with infection and intensity respectively, adjusting for the water catchment area of household water sources, sex, age group, recent praziquantel treatment and village-level open defecation rate. Results 227 participants were included. Overall S. mansoni prevalence was 53.3% (95% CI 46.8-59.8%); median infection intensity was 96 (IQR 48-288) eggs per gram of stool among those infected. Surface water contact was near-universal (97.3%). Prevalence varied markedly between neighbouring villages. No demographic, behavioural, or WASH variable was independently associated with infection. The water catchment area of household water sources was the primary predictor of infection. Conclusions Prevalence substantially exceeded current surveillance estimates, suggesting this population is underserved by existing control programmes. Water catchment area was the dominant determinant of transmission heterogeneity, possibly consistent with Biomphalaria habitat suitability. Water catchment area may represent a valuable and underutilised unit for targeting schistosomiasis surveillance and control in Madagascar.

Electrostatic actuation is an emerging technology for generating tactile sensations on capacitive touchscreens through voltage-induced attractive forces between a fingertip and the surface. However, accurate control of electrostatic attraction during natural touchscreen interactions remains challenging because the applied normal force and sliding speed continuously vary, and their effects on the fingertip-screen contact and resulting actuation strength are not fully characterized. Here, we show how normal force and sliding speed systematically alter fingertip- screen contact area and electrical impedance, and use these measured changes to estimate electrostatic attraction during sliding. Contact area, interaction forces, and electrical impedance were measured simultaneously as participants slid their fingertips across an electrostatic surface under systematically varied normal forces and sliding speeds. These measurements revealed condition-dependent changes in fingertip contact, electrical interaction impedance, effective capacitance, derived effective gap thickness, and electrostatic attraction. We then incorporated these measured contact quantities into a physics-informed, data-driven model based on parallel-plate capacitor theory, in which effective capacitance, apparent contact area, and effective voltage determine the estimated electrostatic attraction. The resulting model links force- and speed-dependent changes in these quantities to electrostatic attraction while accounting for inter-participant variability through a participant-specific scaling factor. These findings provide experimentally grounded guidance for designing electrostatic surface-haptic feedback and future adaptive control strategies under realistic touch conditions.

Background/ObjectivesElymus sensu lato (Poaceae) is arguably the largest and most complex genus in the tribe Triticeae. It includes hybrids and polyploids based on x=7 chromosomes, all including the St genome, forming a valuable genepool for forage grass and cereal breeding. Analysis of chloroplast genome diversity and structural dynamics is critical for resolving maternal lineages, reticulate evolution and biodiversity across this agronomically important complex, refining their taxonomy, conservation and exploitation. MethodsWe sequenced the complete chloroplast genome (plastome) of Elymus ciliaris (4x=2n=28; StStYY genome composition) using ultra-long Oxford Nanopore single-molecule reads and compared it to 76 additional chloroplast genomes representing major St-genome lineages in Elymus s.l. (Pseudoroegneria St; Elymus s.s. StH, StY; Thinopyrum StJ/E; Campeiostachys StYH; Kengyilia StYP). We analyzed structure, nucleotide diversity, inverted repeat (IR) dynamics, and phylogenetic signal. ResultsThe E. ciliaris chloroplast genome was 135,004 bp long (38.3% GC) with a canonical quadripartite structure. Single-molecule reads (n=74) revealed heteroplasmy: two Small-Single-Copy (SSC) orientations at 30%:70% frequency, indicating an inversion polymorphism. Across the Elymus group, comparative analysis of chloroplast assemblies showed high structural conservation but lineage-specific IR-boundary shifts. Kengyilia exhibits exceptional IR expansion. Nucleotide diversity hotspots localize to the large single-copy region, especially in StY lineages. Phylogenies recover a monophyletic St-containing clade but do not delineate genera, reflecting reticulate evolution, with North American/Southeast Asian and Eurasian geographic sub-clades. ConclusionsSingle-molecule sequencing uncovered heteroplasmy with an inversion polymorphism in a single plant of Elymus ciliaris, hidden in short read assemblies. There were no other polymorphisms, as expected for chloroplast sequences (except for technical homopolymer variation). Our analyses showed that a Pseudoroegneria-like St chloroplast genome predominates as the maternal donor across Elymus polyploids. Variable regions and IR dynamics offer strong models for chloroplast genome evolution in reticulate lineages and suggest exploiting plastome variation to complement nuclear biodiversity studies.

Introduction Safe and timely blood availability remains a major global health challenge, especially in low- and middle-income countries. Digital tools may accelerate donor contact, but digital reachability alone does not ensure that people will notice, trust and act on urgent requests to support blood donation efforts. We examined factors associated with anticipated engagement in digitally coordinated urgent blood-donor mobilisation among digitally reachable adults in Ghana. Methods We conducted a cross-sectional online survey from September 2025 to January 2026 across Ghana's 16 regions. Participants were recruited via Facebook advertising and snowball sampling. Factors associated with urgent blood-donor mobilisability were assessed under four criteria: high future-donation willingness; high willingness to install a trusted donation app; high willingness to respond to a trusted urgent-request; and high practical flexibility to leave current activities. Descriptive analyses and multivariable logistic regression examined prevalence and associated factors. Results Among 1,067 participants, 577 (54.1%) met all four criteria. Future-donation willingness (91.8%), trusted-app installation willingness (83.2%) and trusted-request response willingness (82.7%) were common, whereas practical flexibility was lower (66.6%). In the adjusted model, high formal health-system trust (adjusted OR (AOR) 3.95, 95% CI 2.08-7.50), high digital-response readiness (AOR 2.26, 1.66-3.08), previous donation (AOR 1.47, 1.08-2.01), high donation knowledge (AOR 1.42, 1.03-1.97) and willingness to donate to strangers were positively associated with high mobilisability. Women (AOR 0.60, 0.43-0.83), participants reporting a work-schedule barrier (AOR 0.43, 0.29-0.66) and those travelling over 30 min to the nearest healthcare facility at night (AOR 0.66, 0.45-0.96) had lower adjusted odds. Conclusions Digital reachability and stated donation willingness may overestimate the population pool available for emergency donation. Digital blood-donor solutions should consider verifiable health-system requests, account for response readiness and current availability, and connect willing individuals with accessible collection options and transport support where needed.

Despite the well-known health risks of neutron exposures, key gaps remain in understanding neutron-induced molecular responses and identifying reliable biodosimetric markers that distinguish neutrons from photon exposure. We provide the first genome-wide analysis of the human blood transcriptional response to an accelerator-derived fission-like spectrum of neutrons versus photons, evaluating transcriptomic relative biological effectiveness (RBE) and radiation quality-discriminating gene signatures. Whole blood from healthy donors was irradiated ex vivo with X-rays (140 kV, 0-4 Gy, n = 3) or neutrons (0.1-8 MeV, 0-1 Gy, n = 2), incubated for 6 h or 24 h, and processed for RNA sequencing from peripheral blood mononuclear cells (PBMCs). Neutrons were markedly more potent than X-rays at inducing differentially expressed genes (DEGs) at equal doses, showing a peak response 6 h post-irradiation followed by a decline. In contrast, X-rays caused a continuous increase in DEGs up to 24 h (neutrons vs. X-rays at 1 Gy: 1,449 vs. 121 DEGs at 6 h; 996 vs. 621 DEGs at 24 h). A universal p53-centered 34-gene signature, including FDXR, EDA2R, GADD45A, and ZMAT3, showed highly monotonic dose responses (Spearman correlation coefficient {approx} 1) across donors, radiation qualities, and timepoints. Additionally, difference-in-differences analysis identified radiation quality-discriminating genes only at 6 h, with transcriptional convergence observed by 24 h, suggesting a very narrow time window for biodosimetric differentiation. We identified a neutron-specific gene signature driven by cGAS-STING-NF-{kappa}B signaling (RELB, NFKB1, C3, MALAT1) and suppression of B-cell and myeloid identity genes (IGHD, TCL1A, CLEC7A, TLR2), defining a biologically coherent neutron quality index with distinct immunomodulatory effects. For the first time, we assessed neutron RBEs at the gene, pathway, and global transcriptomic levels in a human blood model, reporting a global transcriptomic neutron RBE of 1.30 (95% CI: 1.14-1.49) at 6 h and 1.21 (95% CI: 1.14-1.28) at 24 h, providing a valuable basis for biodosimetry in mixed-field exposure scenarios. Our findings advance the mechanistic understanding of neutron radiation responses and support the development of biodosimetric approaches for mixed-field exposure scenarios.