Anat Sci Int. 2025 Jun 27. doi: 10.1007/s12565-025-00858-x. Online ahead of print.

ABSTRACT

Cerebral microcirculation is a critical infrastructure for brain function, delivering energy substrates and clearing metabolic byproducts. Disruptions in vascular dynamics contribute to neurodegenerative diseases, stroke, and cognitive impairments. Traditional blood labeling methods for fluorescence imaging, such as fluorescent dextran injection, have advanced our understanding of microcirculation but are limited for long-term imaging. In this mini review, we introduce two recently developed molecular genetic techniques, achieved by recombinant adeno-associated virus (AAV)-mediated plasma label expression or genomic knock-in that enable stable, long-term microcirculation imaging. These AAV-mediated methods require only a single systemic injection, facilitating longitudinal imaging of microcirculation in mouse models of disease. We discuss the fundamental design concepts of these approaches and explore their potential applications in systems biology.

PMID:40576868 | DOI:10.1007/s12565-025-00858-x

Appl Biochem Biotechnol. 2025 Jun 27. doi: 10.1007/s12010-025-05293-2. Online ahead of print.

ABSTRACT

One of the main issues that has been receiving great attention globally is the pollution of water resources with dyes, as the chemicals present in them are carcinogenic and harmful to aquatic organisms and humans. Metal oxide nanoparticles synthesized using plant extracts have recently emerged as the best option to address this problem as they provide an eco-friendly and inexpensive approach for a clean water supply. This study describes the photocatalytic degradation of four different types of dyes-methylene blue (MB), rhodamine B (RB), Eriochrome black-T (EB), and methyl orange (MO) using tin oxide nanoparticles (SnO2 NPs) synthesized with ginger (Zingiber officinale) extract. From the results, the SnO2 NPs play a remarkable role in degrading all the dyes even though they require a longer time to reach maximum degradation. This study also describes the biosynthetic preparation of SnO2 NPs, which have a tetragonal structure, prepared using the optimum parameters of 0.02 g mL-1 of ginger extract, 0.5 M of tin (II) chloride dihydrate (SnCl2.H2O), and reaction pH of 5. SnO2 NPs were characterized using powder X-ray diffraction (XRD) to evaluate their crystallinity and composition, Fourier transform infrared (FTIR) to identify the changes in functional groups of SnO2 NPs, and ultraviolet-visible (UV-Vis) spectroscopy to analyze the optical properties of SnO2 NPs through band gap calculation. The photocatalytic activity of SnO2 NPs was then studied through their efficiency in degrading the four dyes. A toxicity test was also carried out in this study using brine shrimp (Artemia salina) to ensure the safety of the synthesized SnO2 NPs made with ginger extract for use in aquatic environments to degrade pollutants. The positive results obtained indicate their potential as a good photocatalyst for water remediation.

PMID:40576720 | DOI:10.1007/s12010-025-05293-2

J Evol Biol. 2025 Jun 27:voaf082. doi: 10.1093/jeb/voaf082. Online ahead of print.

ABSTRACT

Predator-prey systems often feature periodic population cycles. In an empirical system with a heritable prey defence trait, ecological oscillations were previously shown to cause evolution of prey defence on the timescale of the population cycles. In this paper, we develop a phenotypically structured model comprising partial differential equations to investigate the evolutionary dynamics of prey defence during population cycles for a clonally reproducing prey species. We reveal that ecological population cycles not only induce evolutionary oscillations of the mean prey defence trait but also of trait variance. We show that both eco-evolutionary oscillations and stable dynamics lead to high trait variance for a wide range of parameters. For stable dynamics, we show that this is caused by a mutation-selection balance whose impact is larger than in the absence of predators. For oscillatory dynamics, we show that high trait variance is caused by perpetual changes in the direction of selection. Finally, we highlight that switches between stable and oscillatory dynamics depend on the functional form of the cost and efficiency functions of prey defence.

PMID:40576623 | DOI:10.1093/jeb/voaf082

J Exp Bot. 2025 Jun 18:eraf270. doi: 10.1093/jxb/eraf270. Online ahead of print.

ABSTRACT

Copper (Cu) deficiency compromises plant growth and limits crop productivity. Plants respond to Cu scarcity by activating the expression of several microRNAs, known as Cu-miRNAs, which degrade mRNAs from various cuproproteins to conserve Cu. Cu-miRNAs, like most plant miRNAs, associate with ARGONAUTE 1 (AGO1), the primary effector protein of miRNA-mediated gene silencing pathways, whose function is typically modulated by interacting proteins acting as cofactors. However, how AGO1 is regulated and functions under Cu deficiency remains unknown. Here, we searched for AGO1 interactors in Arabidopsis thaliana plants expressing a functional AGO1 protein tagged with the Twin-Strep-tag (TST) polypeptide, grown under Cu-sufficient or Cu-deficient conditions. TST-AGO1 complexes were affinity-purified, and proteins were identified using tandem mass spectrometry. Interestingly, the iron superoxide dismutase 1 (FeSOD1) encoded by FSD1, was enriched in TST-AGO1 complexes purified from plants grown under Cu deficiency. Moreover, fsd1-2 mutant plants showed reduced levels of AGO1 compared to wild-type plants under Cu sufficiency, while both Cu-miRNA-specific and general AGO1 target mRNAs accumulated to higher levels in fsd1-2 plants under both Cu-deficient and Cu-sufficient conditions compared to wild-type plants. These findings suggest that FeSOD1 is essential for proper AGO1 function, and that its superoxide dismutase activity, which mitigates oxidative stress, enhances AGO1 stability, particularly under Cu deficiency.

PMID:40576540 | DOI:10.1093/jxb/eraf270

Plant J. 2025 Jun;122(6):e70300. doi: 10.1111/tpj.70300.

ABSTRACT

Pepper (Capsicum spp.) is a rich source of natural compounds, including primary metabolites essential for plants and influencing human nutrition and taste perception. Although pepper represents an important horticultural crop, the genetic bases underlying the primary metabolism remain largely unclear. Here, we performed a complementary approach for mapping primary metabolites via quantitative trait loci analysis (mQTL) and genome-wide association studies (mGWAS). Using gas chromatography coupled with mass spectrometry we quantified and mapped 80 metabolites, including amino acids, sugars, and organic acids in an interspecific backcross inbred line population and a GWAS panel over three independent trials. We identified 263 candidate genes implicated in 91 robust QTL across studies. Additionally, 28 QTL containing 84 candidate genes were identified with various pleiotropic effects. We further combined agro-physiological characteristics determining their relationships with metabolites, both underlying the quality of pepper fruits. We implemented plasticity analysis to investigate candidate genes causal for metabolic dispersion. Eighty-six genes were identified; among these, a previously reported UDP-glycosyltransferase responsible for capsianosides biosynthesis was found to be associated with a cluster of sugars, organic, and amino acids, which are the main precursors of sensory taste in vegetables. This study provides the first attempt to comprehend the genetic basis of Capsicum primary metabolism, which will further support assisted breeding for fruit quality.

PMID:40576468 | DOI:10.1111/tpj.70300

Int J Radiat Biol. 2025 Jun 27:1-21. doi: 10.1080/09553002.2025.2519561. Online ahead of print.

ABSTRACT

PURPOSE: By expanding the catalog of spaceflight-induced molecular signatures in Caenorhabditis elegans, we aim to identify key molecular features and potential mechanisms underlying space environment-induced health risks to astronauts using C. elegans as a model organism.

METHODS AND MATERIALS: We employed an optimized combination algorithm that integrated two co-expression network analysis methods and four machine learning-based models to identify space environment-responsive genes (SEGs) in space-flown C. elegans. The expression levels and associated biological processes of human orthologues of identified C. elegans genes were further analyzed using data from the JAXA CFE and NASA Twins studies.

RESULTS: A total of 114 SEGs that were implicated in four biological processes, including DNA repair, metabolic process, reproductive and developmental process, and lifespan regulation in space-flown C. elegans. We obtained 19 SEGs as potential indicators associated with health risks of the space environment. Further, the human orthologues of C. elegans SEGs that also exhibited altered expression in the blood samples of astronauts.

CONCLUSIONS: This study provides new insights into the molecular mechanisms behind spaceflight-induced health risks and highlights potential mechanistic targets for preventive measures. The findings suggest a conserved genetic response to space conditions between C. elegans and astronauts, offering valuable targets for mitigating the health risks of space exploration.

PMID:40576322 | DOI:10.1080/09553002.2025.2519561

Microbiol Resour Announc. 2025 Jun 27:e0001425. doi: 10.1128/mra.00014-25. Online ahead of print.

ABSTRACT

We report 26 subsurface sediment and 9 groundwater metagenomes from the Oak Ridge Reservation at Oak Ridge, TN, USA. Samples were collected from various depths and phases (attached vs planktonic) to study subsurface microbial metabolism, the effect of contamination on microbial communities, and differences across groundwater and sediment microbial communities.

PMID:40576316 | DOI:10.1128/mra.00014-25

Bioinformatics. 2025 Jun 27:btaf376. doi: 10.1093/bioinformatics/btaf376. Online ahead of print.

ABSTRACT

MOTIVATION: The rise of transformer-based architectures has dramatically improved our ability to analyze natural language. However, the power and flexibility of these general-purpose models come at the cost of highly complex model architectures with billions of parameters that are not always needed.

RESULTS: In this work, we present CSpace: a concise word embedding of bio-medical concepts that outperforms all alternatives in terms of out-of-vocabulary ratio (OOV) and semantic textual similarity (STS) task, and has comparable performance with respect to transformer-based alternatives in the sentence similarity task. This ability can serve as the foundation for semantic search by enabling efficient retrieval of conceptually related terms. Additionally, CSpace incorporates ontological identifiers (MeSH, NCBI gene and taxonomy IDs), enabling computationally efficient disease, gene or condition relatedness measurement, potentially unlocking previously unknown disease-condition associations.

AVAILABILITY: Full and compressed models are available on Zenodo at https://doi.org/10.5281/zenodo.14781672, while training code, examples, interactive visualizations and experiments are available at https://doi.org/10.5281/zenodo.15125706 and on the GitHub repository.

SUPPLEMENTARY INFORMATION: Supplementary methods (Supplementary File S1) and tables (Supplementary File S2) to showcase the applicability of the embedding space to multiple scenarios.

PMID:40576218 | DOI:10.1093/bioinformatics/btaf376

Front Med (Lausanne). 2025 Jun 12;12:1519216. doi: 10.3389/fmed.2025.1519216. eCollection 2025.

ABSTRACT

BACKGROUND: Hepatitis B virus (HBV) is a globally prevalent pathogen that poses significant public health challenges. Active HBV replication can trigger immune responses that result in liver damage. Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains one of the leading causes of death from a single infectious agent worldwide. Notably, in TB patients with HBV infection and, the incidence of adverse events is six times higher than in those with TB alone, and HBV infection increases the risk of latent TB. However, the relationship between HBV and TB have not been thoroughly investigated.

METHODS: To elucidate the relationship between HBV and TB, we performed an integrated bioinformatics analysis using expression profiling and RNA sequencing data from the GSE83148 and GSE126614 datasets. We identified differentially expressed genes (DEGs) associated with both diseases and analyzed shared biological pathways, key genes, transcriptional regulatory networks, and gene-disease associations. Furthermore, we predicted potential therapeutic agents targeting these shared molecular features.

RESULTS: A total of 35 overlapping DEGs were identified for in-depth analysis. Functional enrichment revealed that these genes are involved in both immune-related pathways and cellular metabolic regulation, underscoring their potential role in the progression of HBV and TB. Protein-protein interaction (PPI) network analysis highlighted four hub genes: CCL2, CD69, EGR2, and CCL20. Additionally, 35 transcription factors (TFs) were predicted to regulate these hub genes. Several candidate drugs, including etoposide, 8-azaguanine, menaquinone, emetine and N-acetyl-L-cysteine, were identified as potential therapeutic options. The DEGs were also significantly associated with other conditions such as pneumonia.

CONCLUSION: This study provides novel insights into the relationship between HBV and TB, offering potential targets for diagnosis and treatment. Our findings may contribute to the development of integrated strategies to manage HBV infection and TB more effectively.

PMID:40575568 | PMC:PMC12198134 | DOI:10.3389/fmed.2025.1519216

RSC Chem Biol. 2025 Jun 19. doi: 10.1039/d5cb00016e. Online ahead of print.

ABSTRACT

In terms of biomass, bacteria are the most successful organisms on earth. This is partly attributed to their tremendous adaptive capabilities, which allows them to sense and rapidly organise responses to changing environmental stimuli. Using complex signalling mechanisms, bacteria can relay cellular information to fine-tune their metabolism, maintain homeostasis, and trigger virulence processes during infection. Across all life, protein phosphorylation represents the most abundant signalling mechanism, which is controlled by a versatile class of enzymes called protein kinases and their cognate phosphatases. For many years, histidine kinase (HK)-containing two-component systems (TCSs) were considered the canonical instruments of bacterial sensing. However, advances in metagenomics has since proven that bacterial phosphorelay is in fact orchestrated by a functionally diverse array of integrated protein kinase types, including Ser, Thr, Tyr and Arg-targeting enzymes. In this review, we provide an up-to-date appraisal of bacterial kinase signalling, with an emphasis on how these sensing pathways are regulated to modulate kinase output. Finally, we explore how selective kinase inhibitors may be exploited to control infections and combat the looming health emergency of multidrug resistant bacteria.

PMID:40575134 | PMC:PMC12189002 | DOI:10.1039/d5cb00016e

Patterns (N Y). 2025 Apr 25;6(6):101239. doi: 10.1016/j.patter.2025.101239. eCollection 2025 Jun 13.

ABSTRACT

Existing antibody language models (AbLMs) are pre-trained using a masked language modeling (MLM) objective with uniform masking probabilities. While these models excel at predicting germline residues, they often struggle with mutated and non-templated residues, which concentrate in the complementarity-determining regions (CDRs) and are crucial for antigen binding specificity. Here, we demonstrate that preferential masking of the primarily non-templated CDR3 is a compute-efficient strategy to enhance model performance. We pre-trained two AbLMs using either uniform or preferential masking and observed that the latter improves residue prediction accuracy in the highly variable CDR3. Preferential masking also improves antibody classification by native chain pairing and binding specificity, suggesting improved CDR3 understanding and indicating that non-random, learnable patterns help govern antibody chain pairing. We further show that specificity classification is largely informed by residues in the CDRs, demonstrating that AbLMs learn meaningful patterns that align with immunological understanding.

PMID:40575131 | PMC:PMC12191730 | DOI:10.1016/j.patter.2025.101239

Brain Commun. 2025 Jun 10;7(3):fcaf231. doi: 10.1093/braincomms/fcaf231. eCollection 2025.

ABSTRACT

Multiple sclerosis (MS) is a chronic inflammatory CNS disease with heterogeneous manifestation. Prognostic markers for early classification of MS are currently under investigation. Higher diagnostic resolution of cerebrospinal fluid (CSF) has the potential to contribute significantly to patient stratification, which should be especially important for a subgroup of patients with high risk to convert to a progressive disease course. This study aimed to determine whether spectral flow cytometry of CSF cells could identify pathogenic CD4+ T cell subset in MS. Using a two-step approach, we designed a marker panel informed by publicly available transcriptomic datasets from early human MS and our own single-cell RNA sequencing (scRNA-seq) in acute and chronic experimental autoimmune encephalomyelitis (EAE), a murine MS model. Notably, chronic ('phase') markers such as Il7r and Ramp3 (associated with memory T cells), Itgb1 (integrin beta-1) and anti-apoptotic genes like Dnaja1, Hsph1 and Jun/AP-1 were enriched in EAE. These markers reflect pro-survival signalling and tissue-residency characteristics, including CXCR6, CD69 and Bhlhe40, which suggest an adaptation of CD4+ T cells towards persistent neuroinflammatory responses in chronic EAE. This phase-specific marker profile highlights CD4+ T cells as both indicators and contributors to disease progression in EAE. Translating these findings to MS datasets, we found an enrichment of phase-specific markers in CSF cells. Spectral flow cytometry in an independent MS cohort revealed distinct memory and effector T cell subsets, indicating unique CSF signatures in MS. This study underscores the heterogeneity and dynamic changes of CD4+ T cells detectable by spectral flow cytometry, enhancing diagnostic resolution of CSF cells and informing more precise therapeutic strategies for MS.

PMID:40574973 | PMC:PMC12199765 | DOI:10.1093/braincomms/fcaf231

Virus Evol. 2025 May 11;11(1):veaf035. doi: 10.1093/ve/veaf035. eCollection 2025.

ABSTRACT

The recombinant FY.4 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variant was first reported in Kenya in March 2023 and was the dominant circulating variant between April and July 2023. The variant was characterized by two important mutations: Y451H in the receptor-binding domain of the spike protein and P42L in open reading frame 3a. Using phylogenetics and phylodynamic approaches, we investigated the emergence and spread of FY.4 in Kenya and the rest of the world. Our findings suggest FY.4 circulated early in Kenya before its export to North America and Europe. Early circulation of FY.4 in Kenya was predominantly observed in the coastal part of the country, and the estimated time to the most recent common ancestor suggests FY.4 circulated as early as December 2022. The collected genomic and epidemiological data show that the FY.4 variant led to a large local outbreak in Kenya and resulted in localized outbreaks in Europe, North America, and Asia-Pacific. These findings underscore the importance of sustained genomic surveillance, especially in under-sampled regions, in deepening our understanding of the evolution and spread of SARS-CoV-2 variants.

PMID:40574750 | PMC:PMC12202047 | DOI:10.1093/ve/veaf035

Bioconjug Chem. 2025 Jun 27. doi: 10.1021/acs.bioconjchem.5c00139. Online ahead of print.

ABSTRACT

Prostate-specific membrane antigen (PSMA) is a promising theranostic target. Different PSMA-targeting small molecule ligands have been FDA-approved or are in development, yet their biological fate at the single-cell level is often unknown. An improved understanding of the cellular distribution of these probes will confer insights into their microdosimetry and guide next-generation theranostic probe development. To enable detailed single-cell pharmacokinetics, it is desirable to have fluorescence affinity ligands that preserve the properties of the native agent. Building upon the structure of the FDA-approved PSMA-617, we synthesized a panel of fluorescent analogs and evaluated their in vitro and in vivo properties. We described a facile solid-phase-based synthesis and optimized the synthesis of the crucial urea pharmacophore. We identified two compounds, PSMA-Lys-DOTA-Cy680 (3) and PSMA-Lys-DOTA-AF647 (4), with similar PSMA binding affinities compared to the parent compound and robust optical imaging properties. Tissue and cellular biodistribution data from imaging can populate microdosimetric and systemic modeling to provide potential insights into future radiopharmaceutical therapy design.

PMID:40574660 | DOI:10.1021/acs.bioconjchem.5c00139

Viruses. 2025 Jun 8;17(6):830. doi: 10.3390/v17060830.

ABSTRACT

In the eastern Brazilian Amazon, the Viseu municipality has almost 70% of its territory deforested. Monitoring viruses from wildlife hosts enables the prevention and control of epizootic events and outbreaks. Seven samples from three marsupials and two rodents were screened by high-throughput sequencing for virome analysis. The three samples from the two Proechimys roberti rodents, one from the liver, one from the brain, and one from a pooled viscera sample, showed the highest results in terms of viral abundance and richness. From these we obtained two strains of a new rodent hepacivirus (RHV), which belongs to a new putative genotype of an unclassified RHV species previously described in Panama and Northeast Brazil. The findings expand the host range of the cited RHV species, imply virus circulation in the study area, and suggest a viral tropism in the liver and perhaps in the brain.

PMID:40573421 | DOI:10.3390/v17060830

Viruses. 2025 May 30;17(6):796. doi: 10.3390/v17060796.

ABSTRACT

Enteroviruses are a group of highly prevalent human pathogens responsible for a wide range of illnesses, ranging from common cold symptoms to life-threatening diseases. A deep understanding of enterovirus biology, evolution, and host interaction is required for the development of effective vaccines and antivirals. Recombinant reporter viruses encoding luminescent or fluorescent proteins have been developed to facilitate such investigation. In this work, using coxsackievirus B3 as our model, we analyze how the insertion of fluorescent reporter genes at three distinct sites within the viral polyprotein affects viral fitness, identifying the most tolerant site for reporter insertion. We then describe a set of experimental workflows for measuring viral fitness, sera neutralization, antiviral efficacy, and recombination using fluorescent reporter viruses. The high homology between different enteroviruses suggests these assays can be readily adapted to study additional members of this medically and economically relevant group of viruses.

PMID:40573387 | DOI:10.3390/v17060796

Pharmaceuticals (Basel). 2025 May 30;18(6):822. doi: 10.3390/ph18060822.

ABSTRACT

Background/Objectives: Acute kidney injury (AKI) remains an unsolved medical problem due to the lack of effective treatments, high mortality, and increased susceptibility to progression to chronic kidney disease (CKD), especially in the elderly. Cellular senescence has been described in AKI, CKD, and aging and has been proposed as a promising therapeutic target. The senolytic drug combination of dasatinib plus quercetin (D&Q) is beneficial in some pathological conditions, including experimental CKD, but there are no data for AKI. Methods: The effect of D&Q combination was tested in folic acid-induced nephrotoxicity (FAN-AKI), a murine AKI model. Results: D&Q pretreatment did not prevent renal dysfunction in the acute phase of FAN-AKI, as determined by serum creatinine and BUN levels at 48 h. Moreover, gene expression of the kidney damage biomarkers Lcn2 and Havcr1, the Cdkn1a gene, which encodes p21, and some genes encoding components of the senescent cell secretome were significantly increased in response to D&Q treatment. The number of senescent p21-positive cells in injured kidneys was similar in untreated or D&Q-treated FAN mice. In addition, D&Q did not prevent the downregulation of the antiaging factor Klotho in damaged kidneys. Conclusions: D&Q treatment was not protective in FAN-AKI, exacerbating some deleterious responses. These results suggest caution when exploring the clinical translation of D&Q senolytic activity.

PMID:40573218 | DOI:10.3390/ph18060822

J Forensic Sci. 2025 Jun 26. doi: 10.1111/1556-4029.70113. Online ahead of print.

ABSTRACT

The distinction between live births and stillbirths is crucial for determining the appropriate legal consequences. Historically, researchers have operated under the principle that an infant's lungs will be filled with air upon death, whereas a fetus's lungs will not. The flotation test for the lungs is the primary method still used in many jurisdictions. However, there are concerns regarding its reliability. For this reason, we conducted a pilot study to evaluate the feasibility of postmortem micro-computed tomography (micro-CT) for differentiating between formalin-fixed aerated and nonaerated lungs. Lung samples from aerated (Group 1) and nonaerated (Group 2) subjects were scanned using micro-CT. We calculated the gas volume percentage (GV%) in each sample. Group 1 (aerated lungs) showed a mean GV% of 9.52 ± 6.77, while Group 2 (nonaerated lungs) showed a mean GV% of 0.58 ± 0.66. These findings suggest that micro-CT can qualitatively and quantitatively detect pulmonary aeration and may serve as a valuable tool in forensic investigations involving suspected neonaticide, feticide, and intrauterine fetal demise.

PMID:40571984 | DOI:10.1111/1556-4029.70113

Can J Anaesth. 2025 Jun 26. doi: 10.1007/s12630-025-02999-z. Online ahead of print.

NO ABSTRACT

PMID:40571892 | DOI:10.1007/s12630-025-02999-z

Nat Commun. 2025 Jun 26;16(1):5403. doi: 10.1038/s41467-025-61071-y.

ABSTRACT

Hepatitis E virus (HEV) is a major cause of acute hepatitis and mainly transmitted faecal-orally. HEV particles present in faeces are naked (nHEV), whereas those found in the blood are quasi-enveloped (eHEV) with a cell-derived lipid membrane. Despite its global health impact, the cellular life cycle of HEV remains poorly understood, particularly regarding the mechanisms of viral entry into host cells. To address this knowledge gap, we develop a high content RNA-FISH-based imaging assay that allows for the investigation of the entry pathways of both naked and quasi-enveloped HEV particles. Surprisingly, we find that integrin α3, previously implicated in nHEV cell entry, is not expressed in the cell types that are most permissive for HEV infection. Instead, we identify integrin β1 (ITGB1) pairing with different α-integrins as the key player mediating nHEV cell entry. Our results indicate that the interaction of nHEV with ITGB1 facilitates entry through Rab11-positive recycling endosomes. In contrast, eHEV particles do not interact with ITGB1 and enter cells using a classical endocytic route via Rab5a-positive early endosomes. The entry of both types of HEV particles requires endosomal acidification and proteolytic cleavage by lysosomal cathepsins, which ultimately results in delivery of the HEV genome to the cytoplasm.

PMID:40571699 | DOI:10.1038/s41467-025-61071-y