Science Advances

Why some surgical participants experience pain that extends beyond the original site of injury while others do not remains poorly understood. Both pain intensity and widespread pain contribute to recovery and quality of life, yet their psychosocial correlates are often examined separately. Using data from two large pre-surgical cohorts--participants preparing for knee replacement or thoracic surgery--we examined associations between sociodemographic and psychosocial factors, pain intensity at surgical and non-surgical sites, and widespread chronic pain. Across cohorts and outcomes, fatigue showed the strongest and most consistent associations with pain intensity and widespread pain, independent of other measured factors. Fatigue also occupied a central position in statistical association networks and accounted for substantial shared variance among multiple psychosocial variables, including sleep disturbance, depression, stress, and socioeconomic disadvantage. Pain at non-surgical sites was strongly associated with widespread pain and frequently accounted for observed associations between surgical-site pain and widespread pain. Together, these findings highlight robust patterns of association linking fatigue, pain intensity, and widespread pain in pre-surgical populations. One Sentence SummaryFatigue is the strongest and most consistent factor linked to how pain intensifies and spreads before surgery.

Nitazenes are driving a wave of overdose deaths in the United States and Europe and often require additional doses of naloxone to reverse. To understand the molecular basis, we conducted a joint experimental and simulation study of three common nitazenes, eto-, etodes-, and protonitazene. Radioligand experiments demonstrated that all three nitazenes display higher receptor affinity and longer dissociation half-lives than fentanyl. Notably, protonitazene dissociates slower than carfentanil and its displacement requires fourfold higher antagonist concentrations. The observed trend in nitazene half-lives is recapitulated by molecular dynamics simulations, which suggest that kinetics is controlled by specific interactions with two receptor subpockets. A newly published cryo-EM structure of fluetonitazene-OR complex confirms the predicted interactions, including a{pi} -hole bond between the nitro group and Tyr1.39, a residue recently shown to modulate OR signaling bias. Our findings suggest slow receptor dissociation as a key factor challenging overdose reversal. The mechanistic insights have implications for understanding opioid toxicity and designing more effective countermeasures.

Mechanistic simulations of blood flow and oxygen exchange showed regions of cortical tissue tolerating substantial increase in local oxygen consumption (CMRO2) before reaching hypoxia (pO2<10 mmHg). The observed robustness in O2 supply was attributed to overcapacity in convective oxygen transport in the pial arterial network combined with a surplus in the number of capillary flow paths. Microcirculatory flux analysis suggests that network induced hemodynamic flow patterns impart intrinsic reserve to protect the brain against perfusion variances or metabolic demand surges during activation. Furthermore, oxygen transport in cortical tissue is characterized by two regimes: in the transport zone -- centered on penetrating arteriole trees composed of a single penetrating vessel connected to the post-arteriole capillary transition zone -- strong diffusion supports high oxygen tension with only modest contribution from capillaries. This regime transitions into the terminal/reactive zone where oxygenation is sensitive to capillary density and perfusion. Quasi-dynamic simulations also enabled reconstruction of the BOLD signal underlying functional imaging. Simulations at single micron resolution further show that age-related reductions in arterial saturation and systemic hematocrit were sufficient to induce hypoxic tissue pockets in the terminal zone at nominal perfusion (CBF) and metabolic activity (CMRO2), and neutrophil adhesion induced capillary flow stalling further exacerbates hypoxia.

Understanding how different populations respond to environmental variation is fundamental to breeding climate resilient crops. In this study, we integrate three diverse georeferenced Cannabis datasets comprising North American feral populations and Eurasian samples (n=909) to resolve population structure, infer evolutionary relationships and quantify adaptive responses to climate across a broad environmental gradient. Phylogenetic analysis rooted to Humulus lupulus shows North American feral populations are more closely related to basal and hemp-type lineages than to drug-type or Iranian populations, a pattern supported by ancestry and PCA analyses. By combining these datasets, we capture a wider range of climatic variation and gain new insight into the adaptive potential of cannabis germplasm. Using environmental genomic selection (EGS), we identified nine bioclimatic traits with prediction accuracies exceeding 0.5 across the combined datasets (12,030 SNPs; 909 samples; training = 191). When analyzing the North American dataset alone, EGS revealed 14 traits with prediction accuracies greater than 0.85 (22,852 SNPs; 760 samples; training = 310). Genomic estimated adaptive values (GEAVs) revealed population specific climatic responses, particularly for precipitation related traits, with northeastern North American populations (Indiana and New York) showing signatures consistent with adaptation to wetter and cooler environments. Climate projections under a high emission scenario (SSP585) indicate that [~]34 % of sampled locations are expected to transition to different Koppen-Geiger climate classes by 2050, with distinct shifts observed among North American and Iranian populations. Genome environment association (GEA) analysis identified replicated temperature and precipitation associated loci across multiple chromosomes. Using genotype-phenotype data, genomic selection for cannabinoid traits revealed that only CBD achieved prediction accuracies exceeding 0.5, consistent with a polygenic architecture extending beyond the CBDAS locus. Collectively, these results demonstrate that feral and landrace Cannabis populations harbor substantial adaptive variation and represent an underutilized reservoir for climate resilient breeding and allele discovery, with relevance for pre-breeding efforts under future climate scenarios.

Phytochemistry is a core player in shaping plant-pollinator networks and pollination services. Yet, little is known about the dynamic evolution of phytochemical traits in response to limited pollinator access, especially concerning chemical diversity. We combined an evolutionary experiment manipulating pollinator access with predictive metabolomics to uncover evolutionary changes in phytochemical traits of Brassica rapa. Our results unveiled chemical changes in both leaf and flower chemistry. Moreover, plants under selection by limited pollinator access showed a decreased chemical richness and diversity and a modulated primary and specialised metabolism, which could be used to predict pollinator access with 88% accuracy. Chemical indices and metabolites responding to pollinator access were associated with variation in flowering time and performance of outcrossing flowers. Our findings provide key insights into the influence of pollinator access on plant chemistry and indicate a risk of pollinator decline and losses of chemical diversity for plant-pollinator network structure and ecosystem dynamics.

Resting-state electroencephalography (EEG) has been proposed as a scalable source of biomarkers for chronic pain, but its clinical potential remains uncertain. To systematically evaluate this potential, we benchmarked nine modeling strategies, spanning conventional machine learning with handcrafted features to state-of-the-art deep learning. Across 72 configurations of signal representations and model architectures, we trained models to predict self-reported pain intensity, using chronological age decoding as a positive control. Pain prediction performance was limited (R=0.15), with the best results achieved by conventional connectivity-based models. In contrast, age was robustly decoded from the same dataset (R=0.53), confirming technical efficacy. These findings indicate that resting-state EEG contains limited information about inter-individual differences in chronic pain intensity, making it unlikely to yield clinically actionable biomarkers in cross-sectional settings. Instead, its potential may lie in intra-individual modeling of pain dynamics, which could advance individualized mechanistic insights and more personalized treatment of chronic pain.

Ecological models using light limitation to explain coral depth distribution have largely disregarded the energetic cost of sustaining photosynthetic activity. Here, we quantified photosystem II (PSII) turnover across a depth-simulated light gradient in a zooxanthellate coral, measuring PSII half-life, D1 protein abundance, and PSII-complex gene expression. Maximum photosynthetic capacity remained stable across irradiance levels while respiration rose and PSII turnover accelerated as a power law, imposing increasing ATP demand at the shallowest depths. Declining D1 protein abundance alongside stable transcript levels demonstrated that this escalating maintenance cost operates through post-transcriptional regulation. Consequently, a decreasing fraction of photosynthetic usable energy is available for translocation to the coral host at high irradiance, as the energy required for PSII repair increases. Integrating these physiological constraints into a bio-optical model revealed that the balance between photosynthetic capacity and its maintenance cost defines an optimal depth, the Photosynthetic Usable Energy Supply (PUES) maximum, where host energetic returns are maximized. This framework provides a mechanistic basis for understanding depth distributions in symbiotic corals and extends as a predictive tool for any photosynthetic organism operating under variable irradiance, including forecasting how environmental degradation contracts viable depth ranges.

Dietary patterns worldwide have shifted toward increased consumption of ultraprocessed foods (UPFs), which has been linked to higher disease burden. One mechanism proposed to impact both their consumption and contribution to metabolic disease is altered post-ingestive metabolic response in comparison to nutritionally similar foods. Here, we recruited 57 healthy-weight 18-45-year-old adults to examine the effects of food processing on postprandial metabolism and brain response. Despite nutritional matching, UPF meals evoked a greater insulinemic and energetic response with attenuated carbohydrate oxidation relative to non-UPF meals. Next, between-condition differences in peak carbohydrate oxidation were associated with mesolimbic and superior temporal gyrus activation in response to food cues. Finally, although food value did not differ between conditions, brain responses correlated with food valuation were positive for non-UPF but negative for UPF in visual cortex and striatum. These findings demonstrate that food processing influences post-ingestive metabolism in a way that could help explain long term health effects and differences in food reward through mechanisms beyond calories and macronutrient composition alone.

The adolescent brain is attuned to social and environmental exploration, allowing behavioral adaptation as experiences shape lasting patterns of morphological organization. Using a convolutional neural network on longitudinal structural MRI data, we assess the early part of this developmental window and derive latent brain representations reflecting patterns of structural variability linked to personal, social, and neighborhood conditions in adolescence. These representations offer a flexible framework for mapping brain-trait associations in adolescence and beyond.

Inorganic phosphate (Pi) availability determines root development and plant performance. In Arabidopsis thaliana, external Pi is sensed by root tips and Pi limitation triggers ER stress-induced autophagy, yet the physiological impact and mechanisms controlling autophagosome biogenesis remain unclear. Here, we identify CAN1 (COMPONENT OF AUTOPHAGIC NETWORK) as a novel plant-specific regulator of autophagosome size in local Pi sensing. CAN1 associates with the plasma membrane-ER tethering protein VAP27-1 and binds to Pi-responsive ATG8 isoforms through conserved interaction motifs. Loss of CAN1 augments ER stress resistance by reducing autophagosome size in Pi-deprived root tips. Our findings establish CAN1 as a determinant of autophagosome size at the ER membrane. Thus, CAN1 functions as a molecular link between Pi-dependent ER stress signaling and fine-tuning of autophagic capacity.

Homeostatic mechanisms protect synapses from destabilizing challenges throughout an organisms lifespan, ensuring stable yet flexible neural network activity. To delineate the molecular basis of presynaptic homeostatic potentiation (PHP), we conducted a comprehensive, in vivo CRISPR/Cas9-based screen of all 16 glutamate receptor (GluR) genes encoded in the Drosophila genome. We first generated a complete expression atlas across larval and adult stages, identifying nine GluRs expressed in presynaptic motor neurons. We then generated null mutants for all 16 GluRs and screened them at the larval neuromuscular junction. While the loss of any single presynaptic GluR did not affect baseline synaptic growth or neurotransmission, our screen revealed a selective and critical requirement for the kainate receptor subunit ekar in the expression of chronic PHP. Further genetic analysis indicates that Ekar functions coordinately with the kainate receptor subunits KaiRID and Ukar within a shared pathway to promote this plasticity. Mechanistically, Ekar acts downstream of active zone remodeling to drive the homeostatic enhancement of presynaptic Ca2+ influx, which is the defining feature of chronic PHP. Together, this genome-wide analysis establishes a definitive functional atlas for the Drosophila glutamate receptome and highlights a specialized, essential role for Ekar in stabilizing long-term synaptic homeostasis.

Volatile pheromones are vital for insect intraspecific communication, yet the genetic basis of homosexual recognition remains elusive. In the bean bug Riptortus pedestris, we show that two previously identified aggregation pheromone components, (E)-2-hexenyl-(Z)-3-hexenoate (E2HZ3H) and (E)-2-hexenyl-(E)-2-hexenoate (E2HE2H), serve as key chemosensory cues for male-male recognition during mate selection. Their biosynthesis is governed by the sex-determination cascade Rpfmd-Rpdsx. The male-specific isoform Rpdsx_M promotes pheromone production in males and induces ectopic synthesis in females upon knockdown of the feminizing switch gene Rpfmd. Knockdown of Rpdsx_M in males abolishes both compounds, prompting wild-type males to court them as if they were females. Metathoracic gland cells act as the production hub. Behaviourally, E2HZ3H or E2HE2H disrupt mating when applied to females: males avoid such females. E2HZ3H reduces female mobility in the presence of the male-derived primary aggregation pheromone tetradecyl isobutyrate (14:iBu), whereas E2HE2H shows no obvious such effect. These differential effects ensure mating accuracy. The discovery of volatile pheromones functioning in male-male recognition and of their synthesis being governed by the sex-determination cascade updates our understanding of mating accuracy in insect chemical communication.

Solid stress shapes tumor growth, invasion, and therapeutic response, yet its physical origin and clinical relevance remain unclear. Here, we develop a mechano-electro-osmotic model integrating metabolic gradients, ion transport, and cellular mechanics to explain residual solid stress emergence in tumor spheroids, common models of solid tumors. We show that solid stress arises predominantly from osmotic cell swelling driven by metabolic deprivation and ion accumulation, rather than proliferation. This mechanism generates a characteristic stress architecture: isotropic compression in the hypoxic core balanced by peripheral tangential tension, causing pronounced cell and nuclear deformation. The resulting nuclear strain provides a mechanical basis for DNA damage and genomic instability implicated in disease progression and treatment resistance. We validate these predictions in breast cancer using MDA-MB-231 spheroids and patient-derived ductal carcinoma in situ lesions, and corroborate them across published spheroid models and in vivo and ex vivo tumors spanning additional cancer types. Our findings link tumor metabolism to clinically relevant mechanical stresses, suggesting opportunities to target osmotic and metabolic pathways to mitigate solid stress and improve therapeutic outcomes.

Sex chromosome differentiation is often accompanied by the evolution of dosage compensation (DC) mechanisms that balance gene expression between autosomes and sex chromosomes, and consequently between the sexes. ZW systems were traditionally thought to exhibit only partial DC, but recent studies in Lepidoptera, Artemia franciscana, and Apalone spinifera suggest a diverse range of compensation strategies, challenging traditional assumptions. While DC often involves chromatin-level regulation, the specific mechanisms in most ZW systems are still poorly understood. To explore these gaps, we generated a genome assembly of Cameraria ohridella (horse-chestnut leaf miner moth, Gracillidae), and combined transcriptomic data from two tissues with CUT&Tag epigenomic profiling targeting both active (H4K16ac, H3K4me3) and repressive (H3K27me3) chromatin marks. Our findings reveal a highly dynamic landscape of sex chromosome evolution in C. ohridella, including the ancestral Z (AncZ), a NeoZ1 formed by fusion of AncZ with an autosome, and a NeoZ2 that likely arose via the fusion of another autosome to the W chromosome. We uncover distinct dosage compensation (DC) patterns across ancestral and neo-sex chromosome regions. On the Ancestral Z, DC involves repression of gene expression in males (ZZ), with a depletion of the active histone mark H4K16ac being observed in this region. In contrast, NeoZ1 regions show upregulation of gene expression in the heterogametic sex (ZW), accompanied by enrichment of H4K16ac. Together, our findings underscore the dynamic nature of sex chromosome evolution and reveal variation in dosage compensation strategies across the Z chromosome, where the NeoZ1 appears to evolve entirely new, Drosophila-like mechanisms rather than co-opting the existing Nematode-like mechanisms of AncZ. We performed simulations that suggest that cooption of ancient DC mechanisms on a neo-sex chromosome is expected when it acts through up-regulation in the heterogametic sex but not when it involves downregulation in the homogametic sex, providing a framework for understanding the different patterns observed between Drosophila and Lepidoptera neo sex chromosomes.

Plant terrestrialization necessitated that a barrage of stressors had to be overcome1. Land plants use an integrated response network in adjusting their molecular physiology to terrestrial stressors2--one of the foremost being UV irradiance. The zygnematophytes are the closest streptophyte algal relatives of land plants3-5, are renowned for their resilience to UV stress6-8, and thus allow to glean key information for inferring the UV response toolkit of the earliest land plants9,10. Throughout streptophyte evolution, specialised metabolism radiated into creating diverse compounds used for responses to environmental challenges, such as sun-shielding compounds and antioxidants11-14. This includes UV-shielding compounds like flavonoids and coumarins but also the land plant specific polymer lignin, giving structural support in vascular plants15; homologs of the underpinning core pathway occur in streptophyte algae16. Here, we exposed the zygnematophyte Mesotaenium to UV-B irradiation and profiled its physiology, morphology, transcriptomics as well as metabolomic features. After UV-B exposure, cells showed rapid photophysiological responses and progressively growing terminal vacuoles. Our transcriptome data capture dynamic changes in gene expression of (i) core downstream responses such as genes homologous to phenol metabolic enzymes, photophysiological homeostats, and DNA repair factors; but also (ii) upstream components featuring key homologs of kinase-mediated signalling cascades, as well as light quality and abscisic acid-mediated signalling components. To scrutinize the acclimatory chassis, we created a metabolite feature database specifically for the Mesotaenium metabolome. Upon UV-B exposure, the metabolome displayed pronounced temporal shifts, with several phenolic features that accumulate along the stress-acclimation kinetics. Overall, we capture a chemodiverse response including various phenolics such as purpurogallin-like, methoxypsoralen-like derivatives and coumarins. Our data establish an integrated model for UV responses in the closest algal relatives of land plants, shedding light on the toolkit that allowed the progenitors of land plants to move out of a protective water column.

Animals rely on olfaction to locate food, mates, and suitable habitats, yet natural odour environments contain thousands of volatile molecules, creating a high-dimensional sensory problem for both nervous systems and the researchers who study them 1-5. For example, a banana emits around 100 individual volatiles4,6. It remains unclear which components of complex odour blends animals have evolved to use as behavioural cues. Here, combining fieldwork, chemical and behavioural analyses, we show across multiple Drosophila species that behaviourally relevant cues can be predicted directly from the statistical structure of natural odour environments. Animals preferentially respond to components that are most distinctive within their natural host odour blends, and therefore most ecologically informative. These cues can be either major or minor blend components. Our results indicate that host-guided olfactory behaviours have evolved to exploit the statistical structure of natural odour environments by selectively targeting the most informative features of odour blends.

Dynamic ionic changes are hallmarks of physiological and behavioral state transitions, including sleep in animals. Although biosensors for specific cellular ions are widely available, real-time monitoring of overall ionic strength in living organisms remains challenging. Here, we present a genetically encoded nuclear translocation ionic sensor (GENTIS) that enables direct visualization of ionic stress in vivo. Using GENTIS in C. elegans, we uncover rhythmic elevations in ionic strength during larval molting transitions that coincide with the lethargus sleep. Cytosolic proton ionic increase through inhibition of v-ATPase is sufficient to induce GENTIS nuclear translocation and evoke behavioral quiescence, characterized by reduced feeding and activation of sleep-active neurons. Apical membrane v-ATPases undergo disassembly during lethargus and under sleep-inducing stress conditions, leading to proton accumulation. Notably, this proton-mediated sleep is suppressed by proton buffering with ammonium. Together, these findings establish GENTIS as a powerful tool for tracking ionic strength dynamics in living organisms and support protons as a physiological driver of sleep.

In their recent study entitled "Ancient DNA reveals the co-existence of domestic horses, donkeys and their hybrids in the prehistorical northwestern China", Li and colleagues (2026) report the genetic identification of three horses, three donkeys and four first-generation hinny hybrids dating to 400-160 BCE from the Mazongshan jade mining site in northwestern China. While a re-analysis of their ancient DNA sequence data confirms the horse and donkey identifications, it indicates that the four putative hinny specimens were, in fact, donkeys. This revision removes the primary evidence originally shown for the presence of hinnies at this site. Therefore, new data from the Mazongshan bone assemblage are required to support the proposed role of hinny hybrids as integral components of trans-regional trade networks during the Late Warring States and Early Han periods.

Animal venoms represent a major source of chemical novelty, yet how venom compounds originate, diversify, and are maintained across deep evolutionary timescales remains poorly understood. This gap is especially pronounced in cephalopods, which evolved venom systems used in predation, defense, and sexual competition, but whose venom genetic architectures, secretory cell types, and venom-producing glands remain largely unexplored. To date, only a single cephalopod venom compound with confirmed paralytic activity and a known primary sequence, SE-CTX from the golden cuttlefish Acanthosepion esculentum, has been described. Here, we reconstruct the evolutionary history, molecular diversity, and glandular localization of SE-CTX-like proteins using a multimodal approach. We identify 29 homologs across 20 squid and cuttlefish species and define a previously unrecognized venom gene family, which we name deca-ctx, specific to decapodiform cephalopods (squids and cuttlefish). Phylogenetic analyses reveal a single origin of deca-ctx followed by gene duplication and lineage-specific diversification, indicating long-term retention of this venom gene. Predicted DECA-CTX protein structures were separated into two clusters and 20 singletons highlighting potentially extensive structural diversity within a single cephalopod venom gene family. Proteomic analysis confirms expression of five DECA-CTX proteins across three species. Our imaging and histological analyses localize deca-ctx expression to specialized secretory cells within squid and cuttlefish venom glands. Together, these findings reposition SE-CTX as part of an evolutionarily and chemically diverse venom system, rather than an isolated venom protein, and establish cephalopods as a key lineage for investigating how new venom genes arise, diversify, and are integrated into functional venom arsenals.

Protein-bound uremic toxins are inefficiently cleared by dialysis and contribute to complications in chronic kidney disease, motivating approaches that target their gut-derived precursors. Here we investigate anaerobic p-cresol metabolism by the environmental denitrifier Thauera aminoaromatica S2, a pathway originally evolved for aromatic pollutant degradation. Proteomic stable isotope probing with 13C-labeled p-cresol reveals strong incorporation of labeled carbon into T. aminoaromatica proteins, whereas parallel incubations with human fecal microbiomes show minimal incorporation, indicating limited intrinsic gut capacity for p-cresol utilization. Label-enriched proteins enable reconstruction of the anaerobic p-cresol degradation pathway and identification of key enzymes synthesized during growth on p-cresol. Moreover, hydrogel-encapsulated T. aminoaromatica remains active during co-incubation with the gut microbiome, achieving complete removal of 0.3 mM p-cresol in less than 10 hours, a timescale compatible with typical intestinal transit in the colon. Together, these findings establish a biochemical basis for repurposing environmental aromatic degradation pathways for gut-localized p-cresol removal.