Open Research Europe

This paper presents a six-stage methodological framework for Convolutional Neural Net-work (CNN)-based cetacean vocalization detection and classification in Passive Acoustic Monitoring (PAM), implemented as the open-source toolkit ai-pam-pipeline. The frame-work is generalizable across species and fully parameterised through a single configuration file, guaranteeing exact experimental reproducibility. Two experiments are reported. Experiment A examines the effect of FFT window length Nfft [isin] {256, 512, 1024} on binary Bottlenose dolphin (Tursiops truncatus) whistle detection using stratified 10-fold cross-validation on an in-domain dataset (Oltremare, 192 kHz) and a cross-domain benchmark (DCLDE 2022). In-domain performance is uniformly high (macro F1{approx} 0.98; Wilcoxon, all p > 0.05). Cross-domain results diverge substantially: Nfft = 256 is significantly superior (p = 0.006, rank-biserial r = 0.89). The mechanism is an upsampling amplification effect: coarser spectral bins produce wider, higher-contrast FM traces after bilinear resampling to fixed image dimensions. This superiority is threshold-invariant: precision equals 1.000 across all configurations and thresholds{theta} [isin] [0.1, 0.9], confirming that the advantage is not an artifact of threshold choice. These findings demonstrate that preprocessing choices -- often treated as secondary implementation details -- can significantly affect cross-domain generalisation. While Nfft serves here as a controlled case study, the framework is designed to enable systematic, reproducible evaluation of arbitrary preprocessing parameters within a unified experimental protocol. Experiment B demonstrates multiclass capability on five T. truncatus vocalization cate-gories (macro F1 = 0.843); inter-class confusion between click trains and burst-pulse sounds reflects biological signal overlap rather than classifier failure.

Pig vocalizations convey information about the emotional states of individuals, varying with arousal and valence. Studies show that different call types reflect distinct emotional contexts and social interactions for the receivers. However, little is known about the brain mechanisms behind the perception of conspecifics vocalizations. This study used BOLD fMRI to explore how pigs brains respond to emotionally varied vocalizations, with the aim to identify activity in regions linked to emotion, reward, and social processing. Eight healthy 2-month-old pigs underwent auditory brainstem response (ABR) testing and BOLD fMRI to assess brain responses to pig vocalizations with different hedonic valence. Sounds were delivered via MRI-compatible earphones, and imaging was performed on a 1.5T scanner. Data were analyzed using voxel-based and ROI-based statistics in SPM12 with small volume correction (SVC). Due to hearing anomalies or MRI artefacts, only 5 pigs were included in the final analysis. Functional MRI revealed that vocalizations activated regions of the auditory pathway and the left amygdala (pFWE at peak < 0.05 after SVC for all), with specific differences between positive and negative sounds. Clusters of activated voxels covering part of hippocampal areas, caudate nuclei and putamen were found with both positive and aversive vocal sounds. Limbic regions, including the amygdala and insula (p<0.05), as well as the right hippocampus after SVC (pFWE = 0.015) were uniquely engaged during the perception of negative conspecific vocalizations, indicating distinct processing based on emotional valence. This study shows for the first time that piglets brain can process and differentiate emotional vocalizations from other pigs, even under general anesthesia. Positive and negative vocal sound playbacks activated distinct brain regions related to hearing, emotion and reward. These findings highlight pigs cognitive and emotional processing of vocal cues. This study is part of a wider research program aimed at developing the fMRI protocol with acoustic stimulation in juvenile pigs.

Clinical metagenomics uses sequencing for culture-independent identification of pathogens directly from clinical specimens. While a number of protocols claim to be pathogen agnostic, sensitivity for RNA viruses is likely lower than for bacteria or fungi, as it requires additional processing steps including conversion to cDNA. Sequence-independent, single-primer amplification (SISPA) was first described in 1991, yet how it preferentially enriches viral molecules has never been described. Here we propose that single-primer amplification exploits the PCR suppression effect, which selectively amplifies longer viral molecules over shorter host-derived cDNA fragments on the basis of size. This model predicts that any upstream processing step that disrupts fragment length will prevent this enrichment occurring. To test this, we systematically compared two adapter introduction strategies - during cDNA synthesis and via tagmentation - followed by single primer amplification, using the ZeptoMetrix Respiratory Panel 2.1 containing 16 RNA and 3 DNA virus strains. SISPA-based approaches recovered all of the viral genomes in the control, whereas using tagmentation to amplify cDNA recovered none. We then spiked the controls into extracted clinical samples and found that SISPA-based methods performed best in all background settings, however in high-background settings no viral genomes were recovered by any approach. Finally, using a modified SMART-9N protocol, we demonstrated that single-primer PCR is critical to overall performance, indicating that direct tagmentation of cDNA and dual-primer PCR should be avoided in protocols for clinical metagenomics where high sensitivity for RNA viruses is critical. These findings demonstrate that library preparation strategy fundamentally determines RNA virus sensitivity and offer mechanistic insights for protocol optimisation with direct relevance to clinical metagenomics.

Rhesus macaques (Macaca mulatta) are widely used as non-human primate models for biomedical research. When housed in captivity, it is essential to provide an environment that supports their natural behaviours; otherwise, they risk developing mood disorders, stereotypies, and other behavioural issues that may lead to physical harm. The objective of this preliminary study was to monitor the behaviour of three aged rhesus macaques ([&ge;] 20 y.o.), relocated from a laboratory to a Rescue Centre for Exotic Animals (Italy), and to assess the impact of novel food enrichments. Behavioural data were collected over 18 weeks, beginning at their arrival, using continuous focal sampling from video recordings. Simultaneously, faecal samples were gathered for cortisol analysis. The study was divided into three phases: a control phase without enrichments, a feeding enrichment phase (divided into two periods), and a final control phase without enrichments. Each phase comprised 900 minutes of observations for each subject. Data were analysed using generalized linear mixed models. Results showed an increase in locomotion during the enrichment and final phase compared to the initial phase. Additionally, a reduction in scratching and body-shaking behaviours was observed in the final phase compared to the initial phase. These findings suggest that implementing an enrichment program can enhance the welfare of aged non-human primates and can be considered a valuable tool in the rehabilitation of non-human primates previously housed in laboratories. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=113 SRC="FIGDIR/small/719840v1_ufig1.gif" ALT="Figure 1"> View larger version (50K): org.highwire.dtl.DTLVardef@152a3a1org.highwire.dtl.DTLVardef@74b53forg.highwire.dtl.DTLVardef@275b21org.highwire.dtl.DTLVardef@1d004d8_HPS_FORMAT_FIGEXP M_FIG C_FIG RESEARCH HIGHLIGHTSO_LIEnvironmental enrichment positively affected activity and stress indicators in aged ex-laboratory rhesus macaques. C_LIO_LILocomotion rates increased while scratching, body-shaking, and cortisol levels decreased. C_LIO_LIEnrichment enhance welfare during rehabilitation, even in older individuals. C_LI

BackgroundSickle cell disease (SCD) is the most common recessive genetic disorder, caused by pathogenic variants of the HBB gene. SCD is associated with a range of clinical manifestations, including vaso-occlusive crises, infections, and severe anaemia, which contribute to increased morbidity and mortality. The frequency of pathogenic alleles is high in Sub-Saharan African countries, with heterozygous carriers reaching up to 25% of the population. Several methods can be employed for molecular diagnostics, with HBB gene sequencing being the most precise. However, access to DNA analyses and sequencing in Low- and Middle-Income Countries (LMICs), where SCD prevalence is high, is limited. Understanding genetic profiles is crucial at both individual and population levels, as it can guide public health strategies and facilitate accurate genetic counselling. AimThis feasibility study aimed to demonstrate that a portable medical genetic laboratory (in suitcases) can be used to genotype individuals for the HBB A, S, and C alleles and their combinations within a few hours outside of a laboratory setting. Methods and resultsWe established a portable medical genetics laboratory capable of DNA extraction and isothermal DNA amplification using a commercially available kit for the A, S, and C alleles of the HBB gene. During one single study day, this portable lab was set up in a room where the Swiss Association of Patients with SCD was holding its annual meeting. We analysed the samples of 27 participants who were aware of their A, S, or C status. We collected buccal swabs and dried blood samples for genotyping. Genotype results for all participants were obtained within five hours after sample collection. In four cases, we observed discrepancies between the buccal swab and blood genotypes; three were resolved upon repeat testing, and one reflected donor chimerism following hematopoietic stem-cell transplantation. ConclusionsThis study demonstrates the feasibility and efficiency of using a portable medical genetics laboratory for rapid genotyping of HBB SCD alleles in community settings.This approach can improve access to molecular diagnostics in resource-limited environments. Such tools have the potential to significantly enhance local capabilities for genetic screening, counselling, and public health planning in regions heavily affected by SCD.

Celiac disease (CD) is strongly associated with specific HLA DQ heterodimers, formed by HLA DQA1 and HLA DQB1 proteins. In particular DQ2.5 (DQB1*02 associated to DQA1*05) and DQ8 (DQB1*03:02 with DQA1*03) are present in virtually all celiac patients. HLA DQB1*02 is considered the main single genetic susceptibility marker and has been reported in 90 to 95% of CD patients. However, the distribution of these alleles may vary across populations, potentially impacting the performance of genetic screening strategies. In this study, we evaluated the prevalence of HLA DQ2.5 and DQ8 genotypes in celiac patients (n = 41) and an unbiased general population cohort (n = 60) from San Luis, Argentina, using a PCR-based genotyping approach. In addition, we assessed the feasibility of a simplified saliva direct PCR protocol for large scale testing. Overall, 95.1% of CD patients carried DQ2.5 and/or DQ8. Notably, 41.5% of patients were DQ8(+)/DQ2.5(-), and 36.6% lacked the DQB1*02 allele, indicating that DQB1*02 based screening alone would have reduced sensitivity in this population. In the general population, 53.3% of individuals carried CD associated genotypes, with a markedly higher prevalence of DQ8 compared to European cohorts. Genotype distributions deviated from Hardy Weinberg equilibrium in CD patients but not in the general population. We show that DQB1*03:02 is a reliable proxy for DQ8, allowing simplification of genotyping strategies, whereas DQA1*05 typing remains essential to discriminate DQ2.5 from other lower risk DQB1*02 carrying heterodimers. We also describe a saliva direct PCR approach showing a performance comparable to purified DNA based assays. These findings highlight the importance of population specific genetic data for optimizing CD screening strategies and foster the development of simplified, cost effective genotyping approaches for large scale applications.

Close to 50% of all bird species are reservoirs of potentially pathogenic fungi, including those listed as priority by the World Health Organization. In Malawi, data on diversity, pathogenic potential, and ecological avian sources of medically important yeast are scarce. A cross-sectional study using a descriptive approach was conducted in Blantyre, Southern Malawi, to characterise medically important yeasts recovered from environments contaminated with excreta/guano from synanthropic pigeons. A total of 20 samples were collected from 4 peri-urban areas, which yielded 71 yeast isolates. To assess the pathogenic potential of the environmental isolates, we compared their phenotypic virulence traits with those of 21 clinical yeast isolates collected from referral hospital laboratories. Pichia kudriavzevii (39%) and Candida orthopsilosis (30%) were the commonly isolated species in the pigeon-guano-contaminated environments. Candida parapsilosis sensu stricto (29%) and Candida albicans (24%) constituted most of the clinical yeast isolates. Half of the species isolated in the pigeon-guano-contaminated environments were also identified among the clinical isolates. A majority of the environmental isolates showed virulence traits similar to or stronger than clinical isolates. The findings underscore the critical need for integrated surveillance under the One Health framework, especially in bird-inhabited spaces close to human settlements.

Common terns (Sterna hirundo) are regionally threatened migratory seabirds that form large breeding colonies during the North American summer months. They are highly vocal and serve as important bioindicators of aquatic ecosystems. Historically, acoustic studies on colonial seabirds have proven difficult due to the dense aggregations of individuals and high rate of call overlap. However, as passive acoustic monitoring (PAM) becomes increasingly common for studying seabird colonies, quantitative descriptions of species vocalizations are needed to accurately interpret behavioral information from colony soundscapes and support automated analysis of large acoustic datasets. This study aims to quantify the vocal repertoire of adult common terns. We deployed AudioMoths to collect acoustic data at a tern colony on Seavey Island, New Hampshire, USA from across the breeding season. Using RavenPro, unique call types were identified through visual and aural inspection of the acoustic data in the spectrogram. For each call, we then extracted measurements of peak frequency (Hz), bandwidth 90% (Hz), syllable duration 90% (s), and total bout duration (s) to quantify the characteristics of each call type. Statistical analyses for acoustic parameters by call type were performed using Kruskal-Wallis tests, followed by post-hoc Dunn tests. Our results demonstrate that each call type is significantly different from another by at least one parameter, with the exception of the kek and kip/tjuk calls. These findings present the first quantitative analysis of common tern vocalizations for North America. By defining temporal and spectral characteristics for multiple call types, this work helps translate colony soundscape into biologically meaningful information about tern behavior and colony dynamics. These descriptions also provide key parameters for developing automated tools to detect and classify vocalizations in dense, noisy colonies. Integrating quantified vocal characteristics with PAM offers a promising approach for monitoring colony activity and behavior while minimizing disturbance relative to traditional methods.

AimsThis study aimed to explore the relationship between gut microbiota composition, obesity, and the effects of a dietary intervention in 50 participants with obesity diagnosis from Antioquia, Colombia. MethodsA single-blind intervention study was conducted, with 25 participants assigned to a control group (CG) and 25 to an intervention group (IG), these last followed a microbiota-enhancing dietary plan for 90 consecutive days. Gut microbiota changes were assessed by sequencing region V3-V4 of 16S rRNA gene and applying the analytical methodology of Biomatest(R) gut health index. Blood biomarkers, including HbA1C, cholesterol, HDL, LDL, triglycerides, and glucose, were measured at baseline and post-intervention. ResultsPrevotella and Succinivibrio were prevalent in the study population. The IG showed significant increases in gut microbial diversity (Shannon index) from baseline to post-intervention. Both groups exhibited significant changes in the Biomatest gut health index, with significant improvements in the IG. Significant correlations were found between dietary intake, blood biomarkers, and microbial abundances, such as the direct association between serum glucose and ultra-processed food intake and between total cholesterol and Dialister. Fish and seafood consumption correlated positively with Akkermansia, while egg intake was associated with higher levels of Desulfovibrio, and Lactobacillus with decreased glycated hemoglobin. The IG experienced a significant rise in Roseburia, a gut health biomarker, while the CG showed higher levels in inflammatory groups like Fusobacteriota. ConclusionsDietary intake significantly influences gut microbiota composition and blood biomarkers. Nutritional programs that improve gut microbiota, as demonstrated by the IG, positively impact gut health in people with obesity diagnosis and may influence healthier dietary choices. These findings support integrating microbiota diagnostics into personalized nutrition strategies, contributing valuable data on Latin American populations. O_FIG O_LINKSMALLFIG WIDTH=195 HEIGHT=200 SRC="FIGDIR/small/25331845v1_ufig1.gif" ALT="Figure 1"> View larger version (48K): org.highwire.dtl.DTLVardef@1652035org.highwire.dtl.DTLVardef@25a04borg.highwire.dtl.DTLVardef@3ed98corg.highwire.dtl.DTLVardef@3caedf_HPS_FORMAT_FIGEXP M_FIG C_FIG

Plasmodium vivax (Pv) infections are developmentally asynchronous and often polyclonal, complicating interpretation of bulk parasite transcriptomes. Here, we analyzed paired in vivo and short-term ex vivo transcriptomes from Ethiopian clinical isolates using stage deconvolution and PvMSP1 haplotyping. Ex vivo maturation modestly increased inferred schizont representation while largely preserving the proportion of trophozoites and gametocytes. After adjustment for parasite stage composition, in vivo and ex vivo transcriptomes remained globally similar, with no genes significantly differentially expressed, indicating the absence of major culture-induced transcriptional response. In contrast, short-term culture reduced multiplicity of infection, contracted within-host haplotype diversity, and non-randomly depleted specific haplotypes, consistent with a clonal bottleneck. In a subset of low-complexity infections, residual expression patterns were clustered by dominant haplotype, suggesting genotype-associated transcriptional heterogeneity independent of developmental stage. Together, these findings indicate that short-term ex vivo culture enriches late asexual stages and selectively filters clones rather than inducing a common transcriptional program. These results shows that ex vivo cultures are reliable way to study gene expression, especially for late stages. However, these needs explicitly model developmental composition and infection complexity when interpreting Pv transcriptomes from natural infections Author summaryMalaria caused by Plasmodium vivax is difficult to study because this parasite cannot yet be grown continuously in the laboratory and infections in patients often contain parasites at different developmental stages and multiple parasite lineages at the same time. In this study, we wanted to understand how much of the parasite gene-expression signal reflects true biological differences, and how much is explained by parasite development or changes that occur during short-term laboratory maturation. We compared parasites collected directly from patients in Ethiopia with matched parasite matured briefly outside the body. We found that short-term culture mainly increased the proportion of later-stage parasites, but after accounting for developmental stage, the overall gene-expression patterns remained very similar. However, culture reduced the diversity of parasite lineages within infections, suggesting that some parasite lineages survive better than others under laboratory conditions. Our findings highlight that natural Pv infections are complex mixtures of parasite stages and lineages. Accounting for this complexity will improve how researchers interpret parasite gene-expression studies and design future studies of parasite invasion, transmission, and survival.

Long-term, automated tracking of group-housed social animals using RFID (radio frequency identification) is a promising approach in ethological neuroscience. However, low-frequency (LF) RFID, while long-established in the field, is constrained by its inherent low data rates, which lead to two critical limitations: (1) compromised spatiotemporal resolution, and (2) the inability to identify multiple tags (animals) simultaneously. To address these limitations, we developed eeeHive, a high-frequency (HF) RFID-based animal tracking system with a fully custom hardware architecture that enables high-speed, multiplexed antenna polling and concurrent multi-tag reading. The polling time per antenna in eeeHive was 5.9 ms, with an additional 8.2 ms read time per tag. We applied the system to track 24 mice for one week, and six common marmosets for seven weeks. The system successfully tracked individuals even within dense clusters, revealing complex behavioral traits characterized by spatial utilization, temporal dynamics, behavioral regularity, and inter-individual relationships. Additional tests with Japanese fire-bellied newts and Nile tilapia juveniles demonstrated comparable tracking performance in aquatic environments. Taken together, eeeHive overcomes the inherent limitations of conventional LF RFID, establishing a powerful HF RFID-based platform for fine-scale behavioral tracking of group-housed animals across terrestrial and aquatic species.

IntroductionMagnetic resonance imaging (MRI) is central to neurological care, yet access remains profoundly inequitable in low- and middle-income countries, especially in rural health facilities where high costs and fragile electricity supply limit services. Ultra-low-field (ULF) portable MRI offers a way to expand access, but deployment in weak-grid settings requires robust affordable power. We characterized the power needs of a 0.064T portable ULF MRI system and assessed the feasibility of a solar-powered MRI-capable facility in a rural Zimbabwean clinic, which we believe to be the first of its kind in the world. MethodsWe measured the power draw of an ultra-low-field MRI session from a portable photovoltaic (PV) battery kit in the UK, quantifying scan, standby and energy use. We then monitored a PV-battery micro-grid supplying a protected circuit at an MRI-capable clinic in Shurugwi, Zimbabwe. Inverter telemetry was used to derive PV generation, load, battery state of charge (SoC) and grid import for working days in October-November 2025, spanning the end of the dry season and onset of the rainy season. ResultsIn the portable configuration, a 64-minute MRI session consumed [~]0.21 kWh, with standby demand of [~]1.44 kWh per 24 hours. In clinic, mean PV generation was 9.10 kWh (SD=1.34) and load 9.91 kWh, with zero recorded grid import and minimum daily SoC typically [&ge;]60%, including during the early rainy season. ConclusionAn affordable PV-battery micro-grid can reliably support ULF MRI and associated research power loads in a rural, weak-grid clinic, offering a reproducible blueprint to narrow diagnostic equity gaps in resource-limited settings.

Holcosus orcesi, the Orces Blue Whiptail, is a Critically Endangered lizard endemic to the upper Jubones River basin in southern Ecuador. Restricted to a narrow elevational range within semi-arid Andean shrublands, it represents one of the few montane members of a predominantly lowland lineage. Here we present the first high-quality reference genome for H. orcesi, generated using Oxford Nanopore Technologies long-read sequencing. The assembly spans 1.68 Gb across only 91 contigs, with an N50 of 76.2 Mb and a BUSCO completeness of 96.8%, making it among the most contiguous and complete squamate genomes to date. Structural annotation predicted 25,682 genes, of which 85% showed homology to known proteins and 45% were assigned Gene Ontology terms. Repetitive elements accounted for 46.3% of the genome, with LINEs representing the predominant class. This genome provides a foundational resource for future evolutionary, comparative and conservation-genomic research of H. orcesi and other mountain reptiles, enabling studies of population genomics, local adaptation, and genomic erosion in isolated populations. By expanding the genomic representation of tropical montane reptiles, this work helps address longstanding phylogenetic and geographic gaps in global biodiversity genomics and provides a foundation for evidence-based conservation of H. orcesi and related taxa.

This work aimed to establish a translationally viable, xeno-free, serum-free platform and protocol for the isolation and expansion of human salivary stem/progenitor cells (hS/PCs) suitable for regulatory qualification and future FDA-approved first-in-human autologous regenerative therapy trials for the treatment of hyposalivation disorders. Parotid gland specimens from non-cancerous regions/tissues were collected from consented surgical patients. Primary hS/PCs were isolated from tissue specimens, cultured in animal-component-free conditions, expanded to produce millions of cells, then enriched for CD44+ stem/progenitor cells by magnetic cell sorting. Normal epithelial purity was assessed using cytokeratins 5/14. Anti-CD133/PROM1 (cancer marker) and anti- fibroblast (clone TE-7) antibodies were used to demonstrate a lack of contaminating cells. Phenotype validation was performed by flow cytometry and immunocytochemistry on both CD44+ sorted and unsorted populations. Senescence-associated beta-galactosidase (SA-{beta}-gal) assays were performed across serial passages (P1-P6). Pluripotency was demonstrated by culture under conditions supporting lineage-specific differentiation. Primary hS/PCs demonstrated consistent expansion and epithelial morphology under serum-free conditions. CD44 expression remained high (>95%) throughout expansion, with negligible detection of CD133 or fibroblast markers, confirming epithelial purity and absence of tumorigenic or stromal contamination. Immunocytochemistry corroborated these expression profiles. SA-{beta}-gal staining revealed only a minor, passage-dependent increase (5-16%) in senescent cells from multiple donors, indicating retention of proliferative potential. Our defined, animal-free culture system supports stable expansion of pure low passage hS/PCs under conditions compatible with good manufacturing practice (GMP).

Background Occupational exoskeletons are used to reduce physical workload and prevent work-related musculoskeletal disorders in physically demanding jobs. Although laboratory studies demonstrate reduced muscle load during simulated manual work tasks, evidence from long-term, real-world implementations remains very limited. The RELAX project aims to investigate the long-term effects of a passive back-support exoskeleton (BSE) during manual order-picking work in a Danish warehouse, focusing on health and socio-economic outcomes. Methods This 18-month controlled in-field intervention study compares outcomes at two warehouse departments: one where workers use a passive BSE and a control group where workers perform work tasks as usual. Approximately 90 full-time workers will be followed during the intervention period with questionnaires, interviews and company-registered performance indicators. Primary outcomes include perceived work intensity and musculoskeletal discomfort, while secondary outcomes include sickness absence, employee turnover, productivity and cost effectiveness. Furthermore, a process evaluation will be conducted based on questionnaires, focus-group interviews, and reported exoskeleton use. Quantitative effects will be analysed using difference-in-difference analysis with generalized linear mixed models to account for repeated measures over time. Employee turnover will be analysed using time-to-event analysis, and qualitative focus-group interviews will be analysed using reflexive thematic analysis to explore implementation processes and contextual factors. Cost-effectiveness and return on investment will be assessed by comparing the investment with potential savings in costs and resource use. Discussion By combining longitudinal quantitative outcomes with qualitative process evaluation, the study seeks to provide ecologically valid evidence on the effectiveness, feasibility and sustainability of occupational exoskeleton implementation. This approach will help clarify whether long-term exoskeleton use improves worker health without compromising productivity and may inform future workplace guidelines and large-scale adoption strategies.

Stress is a major risk factor for mental disorders, and urban living is a key environmental contributor. Nature exposure may promote stress recovery and mental health, but how physiological arousal and subjective stress change across green versus gray space during naturalistic urban mobility is poorly understood. This preregistered study (https://doi.org/10.17605/OSF.IO/HF4RW) employed geolocation-based ambulatory assessment to examine psychophysiological arousal and subjective stress during transitions between urban green and gray environments. Thirty-six healthy urban residents completed a counterbalanced circular walking route in Cologne, Germany, with continuous GPS, cardiovascular, and electrodermal recording alongside ecological momentary assessment of subjective stress, affect, and exertion. Green compared to gray spaces were associated with lower subjective stress and higher affective well-being, with cardiac indices reflecting reduced autonomic arousal during green space exposure. Autonomic changes surrounding environmental transitions persisted beyond the immediate transition window, suggesting that physiological benefits of green space exposure extend into subsequent gray environments. These findings underscore the public health potential of urban green infrastructure for preventing stress-related mental health conditions.

ProblemThe anti-microbial protein granulysin is present in vaginal secretions during the follicular phase of the menstrual cycle but nearly disappears during the luteal phase. The reason for this change is unknown. Method of studyParticipants (n = 23) with regular menstrual cycles collected daily vaginal swabs for granulysin ELISAs. Endocervical cytobrushes, ectocervical biopsies, vaginal biopsies, and PBMC were collected across the cycle to enumerate granulysin-expressing cells by flow cytometry. Cycle phase was determined by daily urinary luteinizing hormone testing and confirmed by serum progesterone levels. ResultsGranulysin levels in secretions were up to 10,000 times higher during menstruation than during the luteal phase (menstruation, median 3,924 pg/mL [IQR 400-17,280]; luteal, median and IQR undetectable [<7.81 pg/mL]). In the endocervical canal, granulysin-expressing cells were much more abundant during menstruation than during the mid-follicular or mid-luteal phases. In contrast, the number of granulysin-expressing cells in the ectocervix and vagina remained stable during the cycle. The most abundant granulysin-expressing cell types in the mucosa were CD8 T cells and NK cells. In a minority of participants, granulysin was consistently detected in luteal-phase swabs; this phenomenon was associated with parity. ConclusionsGranulysin in vaginal secretions is associated with menstruation, which also drives a spike in granulysin-expressing cells in the endocervical canal. This result explains the much higher granulysin levels in secretions during the follicular than the luteal phase. In contrast, immune cells from ectocervical and vaginal biopsies express granulysin independently of the menstrual cycle, indicating their continuous ability to respond to microbial infection.

Understanding plant growth dynamics requires imaging across day-and-night cycles to quantify growth, movement and development in the aerial plant body and to capture the rhythmic nature of these processes. This requires imaging in light during the day and in darkness at night without perturbing plant physiology. Nighttime imaging has typically depended on infrared (IR) illumination, producing monochrome datasets that require specialised hardware and separate analysis pipelines when combined with daytime RGB imaging. Here, we evaluated very low-intensity green (dimG) illumination from standard LEDs as a practical alternative for colour-consistent nighttime imaging and assessed its physiological impact in Arabidopsis thaliana and Lactuca sativa (lettuce). We show that high resolution colour images can be obtained under dimG using low- cost cameras, with sufficient consistency between full-spectrum and dimG images to allow direct comparison and unified image analysis. We show that very low-fluence green light (<0.5 mol m-2 s-1) does not sustain circadian oscillations of gene activity under continuous exposure and does not perturb rhythms when applied during the dark phase of diel cycles. DimG imaging enabled accurate detection of diel leaf movement profiles in Arabidopsis circadian mutants, revealing genotype-specific phase differences under varying photoperiods. In lettuce, dimG pulses and continuous dimG enabled accurate quantification of diel leaf movement without affecting growth, stomatal opening, electron transport rate or chlorophyll content. Motion profiles under continuous dimG mirrored those under darkness. Our findings establish dim green illumination as a cost-effective solution for night-time imaging, simplifying phenotyping workflows with minimal impact on physiology.

The marbled teal (Marmaronetta angustirostris) is a widely distributed but declining waterfowl species, classified as Near Threatened globally and Critically Endangered in Spain. Despite ongoing conservation actions, including ex situ management and population reinforcement programmes, the genomic consequences of long-term captivity, inbreeding, and patterns of functional genetic variation remain unknown due to the absence of a species-specific reference genome. Here, we present the first chromosome-level genome assembly for this species. The genome was generated using PacBio HiFi long reads and Omni-C data, yielding a 1.15Gb assembly with a scaffold N50 of 76.95Mb. A total of 97.16% of the assembly was anchored into 36 chromosome-scale scaffolds, including the Z and W sex chromosomes. BUSCO analysis recovered 99.2% of conserved avian genes. Gene prediction was performed using both ab initio and homology-based strategies, resulting in 16,048 protein-coding genes. This resource provides a foundation for genomewide analyses of inbreeding, demographic history, and adaptive variation, and will support evidencebased in situ and ex situ conservation strategies for this threatened species.

Environmental exposures are increasingly examined in relation to mental health, yet large-scale epidemiological analyses remain constrained by fragmented geospatial data, heterogeneous spatial and temporal resolutions, and privacy-preserving linkage requirements, limiting systematic investigation of multiple environmental domains at the population level. We present environMAP, a harmonised set of analysis-ready environmental exposure layers derived from open, global sources. environMAP spans the built environment, green and blue spaces, light exposure (solar radiation and night-time light), terrain, weather and extremes, and air pollution. We document data provenance, spatial buffers, preprocessing, projection alignment, and metadata, and provide a reproducible workflow for privacy-preserving linkage to cohort residential locations. To demonstrate utility, we linked environMAP to >200,000 adults in the German National Cohort (NAKO) and summarised self-reported lifetime doctor-diagnosed depression across exposure gradients using sex-stratified descriptive analyses. Gradients were interpretable and broadly consistent with prior evidence, supporting feasibility, scalability, and hypothesis generation. The framework is adaptable to other outcomes, cohorts, and regions.