Advanced Science

Spatial transcriptomics enables investigation of tissue organization while preserving molecular and spatial information within intact tissues. However, existing computational methods primarily focus on clustering and batch integration and provide limited characterization of higher-order spatial organization and transferable tissuestate dynamics across heterogeneous biological systems. This study introduces a cross-domain spatial transcriptomic framework centered on recurrence-based latent tissuestate analysis, pathological fragmentation quantification, and transferable representation learning between wound repair and tumor-associated microenvironments. Human spatial transcriptomic datasets spanning cutaneous wound healing, oral squamous cell carcinoma, and head and neck squamous cell carcinoma were integrated within a graph-based latent embedding framework. Recurrence analysis was applied within latent transcriptomic space to characterize spatial organization and remodeling dynamics. A pathological fragmentation index quantified intra-tissue spatial disorganization from recurrence structure. The learned latent embeddings achieved a mean silhouette score of 0.79, demonstrating coherent separation of tissue states. Recurrence analysis revealed progressive restoration of spatial organization during wound remodeling, whereas tumor-associated tissues exhibited increased fragmentation and heterogeneous recurrence structure. Independent single-cell RNA-seq reference atlases demonstrated reproducible multicellular enrichment patterns within latent spatial niches. The proposed framework demonstrates that recurrence-inspired latent spatial analysis may provide biologically interpretable characterization of tissue organization and pathological remodeling across heterogeneous biological systems.

Periodontal disease is characterized by progressive degradation of the gingival extracellular matrix and loss of the physical confinement it imposes on resident stromal cells. In human periodontal tissue, ECM collagen integrity is inversely correlated with facultative nuclear histone acetylation in stromal cells. We hypothesized that matrix stiffness directly coordinates an epigenomic shift in stromal cells. We use a three-dimensional mechanically tunable hydrogel system to independently tune the storage moduli across the mechanical range of healthy and periodontitis-affected gingival tissue. Matrix stiffness drives a genome-wide response in donor-derived human gingival fibroblasts. Matrix-induced confinement leads to an isotropic nuclear geometry and a folded nuclear envelope architecture compared with more permissive, soft matrices. H3K27Ac is suppressed through a stiffness and actomyosin contractility-dependent mechanism. DNMT inhibition in stiff matrices restores the high-acetylation chromatin state with persistent nuclear envelope folding. At the genomic level, stiff matrix confinement drives global CpG methylation gain concentrated at pericentromeric satellite repeats and repeat-dense regions, while collagen synthesis gene promoters and CTCF binding sites are selectively hypomethylated. Non-canonical NF-{kappa}B inflammatory signaling is attenuated through promoter methylation of MAP3K14, and pharmacological NIK inhibition reduces TLR2-stimulated IL-6 secretion in soft-matrix fibroblasts to levels comparable to the stiff condition. These findings identify the gingival ECM as an active epigenomic regulator of stromal inflammatory competence and provide a mechanistic rationale for targeting matrix mechanics to restore stromal homeostasis in periodontitis. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=85 SRC="FIGDIR/small/728299v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@131db0borg.highwire.dtl.DTLVardef@23ba0borg.highwire.dtl.DTLVardef@18b67d9org.highwire.dtl.DTLVardef@14ede81_HPS_FORMAT_FIGEXP M_FIG C_FIG The mechanobiological state of human gingival fibroblasts differs between healthy, stiff extracellular matrices and degraded, soft matrices characteristic of periodontal disease. In a healthy environment, stiff matrices impose physical confinement that enforces an isotropic nuclear geometry, driving dense heterochromatin formation, high global CpG methylation, and reduced histone acetylation. Conversely, the loss of mechanical confinement in soft matrices enables cell spreading and an open euchromatin state, fundamentally rewiring the cellular epigenome to promote non-canonical NF-{kappa}B signaling and chronic inflammation.

Connexin36 (Cx36) is broadly expressed in neurons and serves as the principal protein that forms interneuronal gap junctions (GJs), also known as electrical synapses. Recent high-resolution structures of human Cx36 GJ have revealed crucial electrostatic interactions (ESIs) of charged residues between two docked Cx36 hemichannels at the second extracellular (E2) loops. Despite their structural importance, the mechanistic roles of these ESIs remain poorly understood. To investigate their significance, we systematically designed and tested a series of missense variants targeting key E2 interface residues, aiming to disrupt or modulate the electrostatic landscape at the docking interface. Based on the ESI pairs defined from the crystal structure, our combined computational calculations and dual patch-clamp experiments in engineered HEK293 cell pairs suggest that at least three ESI residual pairs per E2-E2 interface are required to support functional GJ formation. Furthermore, we found that these unique ESIs of Cx36 could play a role in its docking specificity to itself, as they rarely form heterotypic GJs with other brain connexins. Overall, these findings provide essential molecular and functional insights into the mechanisms governing Cx36 GJ formation and partner specificity, paving the way for future therapeutic approaches targeting connexin dysfunction in human diseases.

Three-dimensional (3D) in vitro tissue models are emerging as powerful platforms for studying development, disease, and therapeutic responses, where close monitoring of electrophysiological activity is essential. However, existing probing methods remain limited in accessibility or spatiotemporal resolution for comprehensive electrophysiological mapping of suspended 3D tissues that closely mimic the native environments. Here we introduce a Venus flytrap-inspired bioelectronic mesh system that enables the full spherical enclosure of 3D tissues in a suspended configuration. The system consists of two hemispherical meshes that envelop the tissue, constructed from highly flexible, stretchable, cell-scale ribbons interconnected into a tissue-compliant network with integrated recording electrode arrays. This architecture enables intimate, conformal tissue integration and supports stable electrophysiological recordings over 300 days. The high-resolution recordings allow precise tracking of local dynamics and correlated global signaling, enabling comprehensive assessment of tissue development as well as detailed evaluation of drug responses for disease modeling. Beyond single tissues, the mesh architecture is extended to fully enclose assembloids composed of multiple tissues, enabling characterization of cross-tissue signaling relevant to advanced heterogenous tissue modeling. Furthermore, the system is translated into array-based platforms, demonstrated by a 4x4 array integrating 1024 electrodes, for high-throughput tissue sampling and cross-study analysis. The developed bioelectronic system and integration method provide a broadly applicable platform to advance electrophysiological studies across diverse tissues and organoids.

Laser lithotripsy (LL) is the gold standard for urinary stone management yet maximizing ablation efficiency while maintaining procedural safety remains clinically challenging. Here, we present a visibly transparent, near-infrared (NIR)-absorbing ITO@SiO2 nanofluid irrigation strategy that significantly enhances LL efficiency without compromising endoscopic visibility. By spectrally matching the absorption profile of ITO@SiO2 with the clinical Holmium:YAG laser wavelength, ablation efficiency improved by >200% in the bench-top spot treatments and >100% in the hydrogel kidney model. Mechanistic investigations revealed that the enhanced optical absorption of the nanofluid modifies bubble dynamics and synergistically amplifies photothermal/microexplosion effects and cavitation damage. Importantly, both in-vitro hydrogel and in-vivo porcine kidney models demonstrated a substantial thermal safety margin (maximum temperatures <35 {degrees}C) and excellent acute biocompatibility, with no evidence of thermal tissue injury. Integrating seamlessly into established clinical workflows without requiring stone pretreatment, this strategy offers a highly translatable, safe, and efficient platform for next-generation endoscopic lithotripsy.

Systemic glucose regulation depends on coordinated signaling among metabolically specialized tissues, yet most human in vitro models capture only limited portions of this network. Here, we developed and benchmarked a perfused human six-tissue MPS by combining AnthroHive, a recirculating perfusion platform, with MOTIVE-6, a six-compartment Multiorgan Tissue Interaction Vessel, to culture human gut epithelium, pancreatic islets, liver organoids, adipocytes, skeletal muscle, and midbrain-patterned brain organoids in a microphysiological system. Shared perfusion redirected engineered tissue states toward tissue-aligned metabolic, endocrine, absorptive, contractile, and neural-associated programs while reducing selected isolation-associated stress and remodeling signatures. Under High nutrient conditions, however, multi-tissue interaction shifted liver and islet responses toward inflammatory and nutrient-stress-associated gene expression, indicating context-dependent effects of cross-compartment signaling. Graded nutrient exposure resolved a staged circuit trajectory: Low nutrient conditions supported maintenance-associated programs, Mid nutrient exposure induced compensatory endocrine and anabolic remodeling with declining net glucose depletion, and High nutrient exposure shifted the system toward stress-associated metabolic dysfunction. Under High conditions, metformin and semaglutide produced distinct response modes. Metformin preserved circuit-level glucose handling without increasing insulin or C-peptide accumulation, while semaglutide remodeled gut, brain organoid, islet, and liver organoid transcriptional programs linked to nutrient sensing, epithelial maintenance, endocrine signaling, and neurometabolic state. Together, this study establishes a benchmarked human six-tissue MPS resource, paired with tissue-resolved transcriptomic, shared-media metabolomic, functional, endocrine, and inflammatory datasets, for investigating how tissue interaction, nutrient availability, and metabolic therapies reshape glucose-regulatory networks. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=138 SRC="FIGDIR/small/726943v1_ufig1.gif" ALT="Figure 1"> View larger version (57K): org.highwire.dtl.DTLVardef@181cdfforg.highwire.dtl.DTLVardef@fb211eorg.highwire.dtl.DTLVardef@13b7284org.highwire.dtl.DTLVardef@1db7543_HPS_FORMAT_FIGEXP M_FIG C_FIG Created in BioRender. Trapecar, M. (2026) https://BioRender.com/a4tl7nv

Helicobacter pylori has been classified as a Group 1 carcinogen by the International Agency for Research on Cancer of the World Health Organization and is one of the most well-established risk factors for gastric cancer. Long-term colonization by H. pylori depends on adhesin-mediated attachment to the gastric mucosa, among which the blood group antigen-binding adhesin BabA is a key surface factor involved in host recognition, tissue tropism, and persistent infection. In this study, we established a structure-guided computational design pipeline to develop compact protein binders targeting functionally relevant epitopes of BabA. First, using experimentally resolved BabA-antibody and BabA-nanobody complex structures as templates, we extracted structural contact residues on BabA through heavy-atom contact analysis, thereby defining antibody-recognition epitopes supported by complex-structure evidence. In addition, sequence-based, structure-based, and evolutionary conservation analyses were integrated to identify candidate functional epitope residues with high antigenicity, strong conservation, and surface-exposed features. On this basis, constrained de novo backbone generation was performed around the prioritized epitope regions, followed by amino acid sequence design and structural back-validation of the candidate binders. Candidate BabA-binder complexes were further evaluated using molecular docking, molecular dynamics simulations, and residue-level interface perturbation analysis to assess interface stability, epitope occupancy, and potential binding hotspots. This workflow enables systematic screening of BabA-targeting binders that may compete with antibody-recognized functional surfaces. Although these candidates still require experimental validation, this study provides a transferable computational framework for designing compact protein binders against pathogen adhesins by integrating experimentally resolved complex-structure resources with computational epitope prioritization based on sequence, conformation, and evolutionary conservation, and establishes a preliminary library of BabA candidate binders for subsequent validation and optimization.

Fibrogenesis is essential to wound healing, but aberrant fibrogenesis is a driver of many chronic diseases and cancers. Lysyl oxidases (LOX) play a pivotal role in fibrogenesis by catalyzing the oxidation of lysine residues to reactive aldehydes (allysine) in collagens and elastin, resulting in the crosslinking and excessive deposition of these extracellular matrix components. Currently, rapid and robust histological assays to visualize the spatial distribution of LOX activity are lacking, hindering the precise validation of anti-fibrotic therapies. Here, we present a histological fluorescent staining method to visualize fibrogenesis (active fibrosis) and LOX activity in tissue sections utilizing a bioorthogonal tag and a click reaction with a turn-on fluorophore. Notably, requiring only two commercial reagents, this protocol can be completed in under two hours and is compatible with other imaging modalities, including second-harmonic generation and immunofluorescence staining. We validated this method across various healthy and fibrotic mouse and human tissue specimens.

Abdominal aortic aneurysm (AAA) is a life-threatening vascular disease characterized by chronic inflammation and immune dysregulation, with lesional macrophages playing a pivotal role in disease progression. However, effective and safe delivery of immune modulators to macrophages at the site of AAA remains a major clinical challenge. To address this unmet need, we report a nature-inspired nanodisc platform based on high-density lipoproteins for targeted delivery to lesional macrophages, further engineered with a multi-component targeting strategy incorporating an aneurysm-homing peptide and phosphatidylserine lipids. Nanodiscs encapsulating an anti-inflammatory protein kinase R-like endoplasmic reticulum kinase (PERK) inhibitor remarkably attenuated progression of established AAA in an elastase-induced mouse model. Using a combination of in vivo biodistribution and immune profiling approaches, we demonstrate that nanodisc-assisted PERK inhibitor delivery selectively reprograms the local immune microenvironment and attenuates pathological inflammation in AAA disease models. Notably, a single administration achieves sustained therapeutic efficacy with favorable safety profiles, effectively limiting the progression of established AAA in a clinically relevant setting. This work presents a new avenue of designer nanomedicines for targeted immunomodulation and maybe broadly applicable for a wide range of vascular and immune-mediated pathologies.

We present a compact pump-and-probe mid-infrared Optothermal Spectrometer (OTHES) equipped with Spatial Probing and Autocorrection (SPAC) optimized for robust intravital application in humans. SPAC-OTHES facilitates alignment stability and spectral comparability across different measurement sessions involving different skin types. Contrary to state-of-the-art, SPAC-OTHES uses camera-based beam detection and an auto-calibration mechanism that enables ca. 73% better spectral reproducibility in intravital measurements in human volunteers than non-calibrated readouts. Moreover, SPAC-OTHES has the potential to lower the glucose quantification error, as demonstrated here in artificial skin phantoms, where an improvement of 52% compared to conventional diode-based detection was observed. The compactness of OTHES, combined with reliable SPAC-readout, has the potential to accelerate commercialization and broad application of biosensors based on mid-infrared spectroscopy.

The subarachnoid space is critical to cerebrospinal fluid (CSF) circulation and waste clearance, yet its flow organization in humans remains poorly characterized due to the lack of methods for direct, quantitative measurement of ultra-slow flow. Here we introduce slow-flow-sensitized phase-contrast imaging (SOPHI), which enables noninvasive quantification of ultra-slow flow velocities (e.g., [~]100 m/s) and directionality at high spatiotemporal resolution, allowing whole-brain mapping of complex CSF circulation in the human subarachnoid space. We found brain-wide, spatiotemporally coherent flow patterns characterized by strong cardiac-driven oscillations. Flow dynamics were closely coupled with vascular anatomy, exhibiting higher velocities and earlier responses in subarachnoid spaces near major arteries and a spatiotemporal propagative pattern to distal spaces. We further identified localized flow pathways associated with potential CSF efflux in ventricles and key subarachnoid regions. Together, SOPHI reveals a previously undercharacterized, highly organized CSF flow system, providing a macroscopic view of human CSF circulation and a framework for investigating brain fluid transport.

Astrocytes play a significant role in neuroprotection by internalizing neurodegenerative aggregates and facilitating their degradation. Recent studies indicate that -Synuclein (-SYN) protofibrils promote the transfer of pathogenic aggregates and dysfunctional mitochondria between astroglia via tunneling nanotubes (TNTs), which enhances cell survival and resistance to apoptosis. However, the underlying mechanism of TNT-driven apoptosis resistance remains unclear. We find that -SYN protofibrils induce aberrant mitochondria with decreased membrane potential ({Psi}m) and promote dynamic actin remodeling by relocating phosphorylated focal adhesion kinase (pFAK) to the nucleus, which triggers TNT formation in human astrocytoma cell lines and primary murine astrocytes. The important novel finding of this study is that pFAK in the nucleus co-localizes with Nanog, a crucial transcription factor for preserving stemness, and the interaction between pFAK and Nanog is critical for promoting p53 degradation via Mdm2-mediated ubiquitination and upregulating autophagy, thereby supporting the survival of astroglia exposed to toxic -SYN protofibrils. ROCK inhibitor y-27632 also drives TNT-formation via pFAK translocation to the nucleus, colocalizes with Nanog, and enhances stemness-related gene expression. Inhibiting TNT with the actin depolymerizing agent cytochalasin-D prevents pFAK co-localization with Nanog in the nucleus and fails to protect cells from -SYN-induced apoptosis. Nanog knockdown does not degrade p53 and hinders cell rescue from apoptosis. Furthermore, these transient TNTs transfer mitochondria to adjacent cells, potentially helping maintain metabolic stability. This study reveals that the TNT formation pathway promotes pFAK-Nanog interaction in the nucleus, leading to p53 degradation, which protects astroglia against -SYN proteotoxicity and prevents apoptosis. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=194 SRC="FIGDIR/small/727344v1_ufig1.gif" ALT="Figure 1"> View larger version (57K): org.highwire.dtl.DTLVardef@18d8675org.highwire.dtl.DTLVardef@76a383org.highwire.dtl.DTLVardef@e92202org.highwire.dtl.DTLVardef@1b7ee4b_HPS_FORMAT_FIGEXP M_FIG C_FIG

Precise activation of ion channels enables fine-tuned control of neuronal excitability, providing a powerful strategy for dissecting and modulating neural circuits. Although optogenetics offers high spatiotemporal neuronal manipulation with cell-type specificity, its reliance on visible light (400-650 nm) limits tissue penetration to millimeter depths, restricting applications in deep-brain stimulation. Here we report a near-infrared-II (NIR-II)-sensitive calcium channel, HaloNeu, created by genetically fusing a circularly permuted HaloTag (cpHaloTag) to the thermo-sensitive transient receptor potential vanilloid 1 (TRPV1) and covalently conjugating NIR-II photothermal nanotransducers (HPN). The HaloNeu enables non-invasive neuromodulation at depths up to 1.0 cm at ultralow laser power ([~]60 mW/cm2) and up to 5.0 cm under the safe exposure limit ([~]1 W/cm2) with 1064 nm laser illumination. Remarkably, HaloNeu maintains stable, on-demand neuron-specific modulation for over two months in vivo, providing sustained activation of ventral tegmental area (VTA) circuits and effective alleviation of Parkinsonian symptoms in mouse models. These results establish HaloNeu as a robust and versatile platform for cell-type-specific, deep-tissue, and chronic neuromodulation, with broad implications for neuroscience and neurotherapeutics.

Postoperative cognitive decline (POCD) after coronary artery bypass grafting (CABG) is increasingly conceptualized as a system-level disturbance of large-scale brain coordination rather than focal dysfunction. Here, we propose a multiscale neural engineering framework that combines static and dynamic information-theoretic connectivity with graph-theoretical analysis to characterize postoperative network vulnerability and its association with cognitive outcome. Resting-state fMRI was acquired in 14 male CABG patients at an early postoperative baseline (BL) and at 3-month follow-up (FU). Cognitive outcome at follow-up was assessed with the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS), classifying 7 patients as POCD (RBANS < 80) and 7 as NO POCD. Functional connectivity between 32 brain regions, grouped in 8 resting-state networks (RSN), was estimated using mutual information (MI; static dependence) and mutual information rate (MIR; dynamic information exchange), each computed with parametric Gaussian (linear) and model-free k-nearest neighbor estimators. Pairwise connections were validated via surrogate testing, and group differences in longitudinal connectivity change ({Delta} = FU-BL) were assessed with permutation tests at global, intra- and inter-RSN scales. Graph metrics were computed on statistically thresholded weighted networks and related to RBANS using permutation-based Spearman correlations. POCD was not associated with a uniform reduction in connectivity but with a structured pattern of network reorganization. Static connectivity showed widespread alterations, particularly within higher-order associative systems, including salience, dorsal attention, and default mode networks. Dynamic connectivity did not exhibit global group differences but revealed selective, network-specific alterations in temporal information exchange. Longitudinal analyses showed that better cognitive outcomes were associated with increased global efficiency and density and reduced modularity and small-worldness, indicating a greater brain integration. In contrast, poorer outcomes were associated with increased segregation and higher betweenness centrality, suggesting greater reliance on hub-mediated communication. Linear measures captured more widespread connectivity changes, whereas nonlinear estimators revealed more selective alterations in dynamic information flow. Combining static and dynamic information measures with complementary estimators and surrogate-validated graph analysis reveals dissociable signatures of postoperative network dysfunction. POCD is characterized by impaired restoration of distributed integration and a progressive shift toward hub-dependent communication, suggesting that large-scale integrative vulnerability may constitute a candidate biomarker of cognitive resilience after cardiac surgery.

Hematoxylin and eosin (H&E) has been the fundamental method for visualising tissue morphology. Recent advances in tissue clearing and microscopy have enabled the observation of tissue morphology in 3D, but incomplete penetration of nucleic acid dyes has remained the bottleneck. To address this, we develop a new staining chemistry called Cyclodextrin and Organic solvent-assisted deep Labelling of ORgans in 3D (COLOR-3D), which attains the best penetration depth and homogeneity among state-of-the-art methods. We also demonstrate the scalability of COLOR-3D and its compatibility with other staining modalities. To bridge the gap between 3D histology and its wider application in biomedical research and histopathology, we develop a computational pipeline to convert 3D fluorescence images into bright-field H&E images, enabling the creation of a 3D atlas of both normal tissues and pathological specimens. Apart from qualitative observation of tissue morphology, COLOR-3D also enables quantitative analysis for studying biological phenomena. In the mouse liver, we discover rare populations of tetranuclear hepatocytes as well as m16n, t4n and t8n hepatocytes. We also propose the first structural model of the liver lobule based on 3D histology. With a more complete penetration, we reveal the following aging-related changes in tissue microarchitecture, including an increase in extreme nuclear polyploidy, and the disruption of vasculature and portal triad.

Engineering biomimetic extracellular matrices that isolate specific biochemical cues is essential for understanding how matrix chemistry regulates tumor cell behavior and therapeutic response. Aberrant sulfation due to proteoglycan expression is a hallmark of lung tumor matrices, yet its functional impact is difficult to study using conventional materials where mechanical and biochemical variables are coupled. To address this, mechanically matched sulfated alginate hydrogels are engineered to mimic the elevated sulfated glycosaminoglycan (sGAG) content of malignant ECM, enabling sulfation to be examined as a single, tunable variable. Within this system, ECM sulfation is shown to enhance tumor cell proliferation, promote oxidative and mitochondrial stress tolerance, suppress apoptotic signaling and attenuate the efficacy of cisplatin, gemcitabine and paclitaxel. Sulfated matrices preserve mitochondrial membrane potential, limit ROS accumulation, shift apoptotic gene expression toward a survival-favoring profile, selectively upregulate ABCB1-mediated efflux and modulate drug response through the PI3K/Akt-ABCB1 signaling axis. Functional inhibition of PI3K and ABCB1 uncovers drug-specific dependencies while dual pathway targeting completely restores chemotherapeutic sensitivity. These findings identify ECM sulfation as a potent regulator of stress adaptation and therapeutic efficacy in lung adenocarcinoma and underscore the importance of biomimetic ECM design in controlling tumor cell fate and drug response.

Anxiety disorders are highly prevalent and often refractory to existing treatments, motivating the development of alternative neuromodulatory strategies. Peripheral bioelectronic approaches targeting the gut-brain axis such as vagus nerve stimulation (VNS) demonstrate that modulation of visceral afferent pathways can influence central emotional circuits. Gastric electrical stimulation (GES) is a clinically established therapy for gastrointestinal motility disorders. While the stomach is densely innervated by vagal afferents, the effect of GES on anxiety-related behavior has not been systematically examined. We sought to identify neural pathways engaged by GES and effects of continuous chronic GES on behavior. To do this, we developed a gastric stimulation platform for rodents, a fully implantable, untethered system enabling chronic neuromodulation in freely moving rats. We combined this with cross-species whole-brain activity mapping in mice to interrogate circuit-level mechanisms. Using open field and elevated plus mazes, together with machine-learning-based behavioral tracking and multivariate modeling, we show that acute GES induces a robust, context-dependent anxiogenic phenotype characterized by reduced exploration and increased freezing, particularly in novel open-field environments. In contrast, chronic GES produces a divergent post-stimulation behavioral profile marked by enhanced exploratory behavior relative to acutely stimulated animals, indicating temporally dynamic reorganization of anxiety-related behavior. Principal component analysis and hierarchical clustering further revealed that stimulation reshapes the multivariate structure of behavioral features rather than shifting animals along a single anxiety continuum. Whole-brain c-Fos mapping revealed anatomically distributed modulation of limbic-cortical networks following gastric stimulation, including suppression of ventral medial entorhinal cortex, excitation of nucleus tractus solitarii and heterogeneous recruitment of amygdalar and hippocampal subregions. These circuit-level patterns align with the behavioral dissociation between contextual exploration and explicit threat avoidance, providing convergent cross-species evidence that gastric stimulation engages distributed anxiety-related networks. Together, these findings establish the first freely moving behavioral model of chronic gastric neuromodulation, demonstrate temporally dynamic and context-sensitive effects on anxiety-like behavior, and provide systems-level validation that the stomach can serve as a viable peripheral access point for modulating central emotional circuits.

Amyloid oligomers have been widely implicated as primary cytotoxic intermediates; however, their selective and scalable production remains challenging due to rapid fibril amplification. Here, we demonstrated that sustained axial rotation enables programmable mechanochemical control over amyloid pathway selection without the use of chemical additives. Using a thermal axial rotator, native monomeric hen egg-white lysozyme was incubated at 60 {degrees}C under quantitatively tunable rotational speeds, imposing defined centrifugal forces and wall-associated shear that restructured the hydrodynamic boundary conditions. A discrete transition emerged near 600 RPM, separating the two distinct assembly regimes. Below this threshold, aggregation followed a fibril-amplifying pathway characterized by elevated {beta}-sheet content and elongated fibrillar morphologies. Above this threshold, fibrillar growth was strongly attenuated and oligomer-dominant assemblies predominated. Spectroscopic analyses and atomic force microscopy revealed that axial rotation redistributes the amyloid assembly states rather than simply suppressing aggregation. Functionally, the RPM-defined assemblies exhibited kinetically distinct seeding behaviors and induced divergent cytotoxic phenotypes in SH-SY5Y neuroblastoma cells. These findings establish axial rotation-mediated hydrodynamic boundary control as a scalable, chemical-free strategy for reprogramming amyloid assembly pathways and producing oligomer-rich assemblies with well-defined structural and functional properties.

Flexible organic small molecules can assemble into supramolecular biomaterials whose properties are intrinsically governed by their crystal structures, yet experimental structure determination remains difficult to scale during molecular modification and materials optimization. Crystal structure prediction (CSP) provides a potential solution, but its prospective power is limited for flexible molecules owing to incomplete conformational sampling and the difficulty of identifying experimentally realized structures from large candidate sets. Here, an SE(3)-equivariant deep-learning workflow, SE3CSP, is developed for organic crystal structure prediction. By learning molecular conformations, unit-cell parameters and packing patterns from experimentally resolved crystal structures and integrating these predictions with MACE-based structure optimization, SE3CSP establishes an end-to-end pipeline from two-dimensional molecular representations to three-dimensional crystal structures. Using nucleosides as representative flexible self-assembling building blocks, SE3CSP achieves an overall prediction accuracy of [~]57%, substantially outperforming the benchmark MACE-based Genarris 3.0 workflow ([~]14%). Furthermore, a prospective prediction strategy is developed in which an SE3CSP-predicted density window ({+/-} 0.15 g/cm3) is applied prior to energy ranking and structural deduplication, enabling all experimentally realized structure to be consistently ranked within the top 1-2% of all generated candidates. Beyond structure prediction, SE3CSP-derived energy landscapes provide insight into potential single-crystal-to-single-crystal transformations and enable the identification of a nucleoside supramolecular material with dynamic breathing porosity, which is further developed as an adsorptive platform for inflammatory mediator removal with excellent anti-inflammatory performance and biocompatibility. These results establish SE3CSP as a practical framework for prospective CSP and highlight its utility in guiding the discovery and design of bioactive self-assembled materials.

Anti-staphylococcal lytic agents, such as lysostaphin (LSN), a glycyl-glycyl (Gly-Gly) peptidase, have long been considered for the management of chronic and complicated bacterial infections typically resistant to conventional antibiotics, but their use is restricted by poor pharmacokinetic properties. We generated half-life extended lysostaphin constructs by fusing the lysin - either alone or chimerized with an additional enzymatic cysteine, histidine-dependent amidohydrolases/peptidase (CHAP) domain - to the human IgG1 Fc fragment. The Fc-CHAP-LSN constructs retain high potency against Staphylococcus aureus and coagulase negative staphylococcal strains in vitro and are efficacious in S. aureus ex vivo biofilm models and in vivo sepsis models. A detailed investigation of the Fc-CHAP-LSN mode of action revealed that upon binding to the bacterial cell, but not in solution, LSN mediates its own release by cleaving the Gly-Gly CHAP-LSN linker. The bactericidal activity of Fc-CHAP-LSN is driven by the LSN-catalyzed and target cell-dependent release of free LSN. The half-life extended lysin acts as a pro-drug, unveiling a novel mechanism of targeted release and an alternative approach to half-life extension.