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A phase oscillator model of cell cycles reveals nuclear density control in a branched fungal network

Maintaining an appropriate nuclear-to-cytoplasmic ratio is essential across cell types for physiological function, and mechanisms of size control have been extensively studied in mononucleate cells. Much less is known about how comparable control is achieved in cells where many nuclei share a common cytoplasm, which are seen in many contexts including muscle, placenta, and filamentous fungi. The filamentous fungus Ashbya gossypii forms a branching mycelial network in which individual nuclei divide asynchronously, while the number of nuclei per cell volume (the nuclear density) is tightly controlled. How global regulation of nuclear density coexists with local cell cycle asynchrony remains unclear. To address this we model nuclei as a dividing population of phase oscillators within a branching cell network and parameterize the model with measurements from Ashbya cells. The model demonstrates that asynchrony is required to prevent large density fluctuations that would result from synchronous division, and that introducing a nuclear density checkpoint to the cell cycles leads to synchrony if it is the only mechanism of density control. We find that coupling branch formation to nuclear density both stabilizes nuclear density and prevents the emergence of synchronous cycles. Supporting these predictions, we demonstrate that mutants with branching defects and increased cell cycle synchrony display greater variability in nuclear density. Our results indicate that asynchronous nuclear cycles together with density-responsive branching maintain a constant nuclear density, revealing a strategy for regulating the nuclear-to-cytoplasmic ratio in large multinucleate cells.

The effect of a body shape index on physical fitness index is more pronounced in boys than in girls: Evidence from a cross-sectional survey based on Tibetan adolescents aged 13–18 years in high-altitude areas of China

Background China is an important high-altitude region in the world, mainly inhabited by the Tibetan population. This study was conducted to understand the association between a body shape index (ABSI) and physical fitness index (PFI) among Tibetan adolescents in high-altitude areas of China. This study provides a reference for the improvement of physical fitness and the formulation of public health policies among Tibetan adolescents in high-altitude areas of China. Methods Using a stratified whole-group sampling method, 3819 Tibetan adolescents were selected from the high-altitude region of Ganzi, Sichuan, China, and tested on 8 items, including height, weight, waist circumference, grip strength, standing long jump, seated forward bend, 50-meter run, and endurance run (1000m/800m). ABSI and PFI were calculated, and Kruskal-Wallis H was used to compare PFI between groups. Linear regression analysis was used to analyze the correlations that existed between ABSI and PFI. Results The ABSI of Chinese Tibetan adolescents aged 13–18 years was (0.09 ± 0.01). Among them, ABSI was (0.09 ± 0.01) for boys and (0.09 ± 0.01) for girls, with no significant difference in comparison ( t  = 1.594, P  > 0.05). PFI [M(P 25 ,P 75 )] for Tibetan adolescents aged 13−18 years was −0.37 (−2.31,1.48), among them, PFI for boys was – 0.24 (−2.41,1.63) and the girl’s PFI was −0.46(−2.20,1.35), with no significant difference in comparison ( Z value = −0.878, P  > 0.05). Overall, the comparison of PFI between the different ABSI groups (A, B, C, and D) was statistically significant ( P  < 0.01) in all age groups except for the 17-year-old age group where there was no significant difference ( P  > 0.05). Conclusion The ABSI level of Tibetan adolescents in high-altitude areas in China is low and negatively correlated with PFI. Compared with girls, the effect of ABSI on PFI was more obvious in boys. It is suggested that in the future, Tibetan adolescents in high-altitude areas should control the occurrence of waist circumference and overweight obesity and increase physical exercise to promote the physical fitness level of Tibetan adolescents.

The combinatorial innexin code of heterochannel electrical synapses governs synaptic function and is maintained by distinct cellular mechanisms

Diversity in the molecular composition of intercellular gap junction channels, the functional units of electrical synapses, determines their physiological properties. Yet the mechanisms by which neurons coordinate the use of multiple coexpressed gap junction proteins—connexins or innexins—to establish selective connections with distinct synaptic partners remain largely unknown. Using the posterior mechanosensory circuit in Caenorhabditis elegans, we found that individual electrical synapses can form by clustering molecularly distinct gap junction channel types composed of three different innexin proteins: INX-1, UNC-7, and UNC-9. This previously uncharacterized combinatorial configuration, which we term heterochannel synapses, enables molecularly distinct gap junction channel types to collaborate functionally to regulate posterior touch sensory behavior and enhance functional robustness. We show that the synaptic trafficking of these molecularly distinct channel types within a heterochannel synapse is independently regulated by specific and conserved kinesin motor proteins, while distinct molecular pathways involving channel type-specific retrograde kinesins control their turnover. These independent, channel-specific regulatory mechanisms allow for individual synapse-level alterations in the combinatorial innexin code of heterochannel synapses in response to altered environmental conditions, providing a mechanism for electrical synapse plasticity. Finally, we present evidence and functional significance of heterochannel electrical synapses in C. elegans locomotory circuits. Altogether, our findings demonstrate a combinatorial heterochannel organization of electrical synapses, their functional significance, and the regulatory mechanisms that govern them.

Associations of demographic, socioeconomic, lifestyle factors and comorbidity with accelerometer-measured physical activity in adults with cardiovascular diseases

Cardiovascular diseases (CVDs) are a leading cause of morbidity and mortality, and physical activity (PA) is central to secondary prevention. However, factors associated with PA among individuals with CVDs remain poorly understood, with few studies using accelerometer-based methods. This cross-sectional study examined the associations between accelerometer-measured PA and demographic, socioeconomic, lifestyle factors, and comorbidity in a large Swedish cohort of individuals with CVDs. Participants aged between 50 and 64 years with a confirmed diagnosis of angina pectoris, myocardial infarction, atrial fibrillation and flutter, heart failure, or stroke were included. Moderate-to-vigorous physical activity was measured using hip-worn triaxial accelerometers over seven consecutive days and categorized into tertiles of low, medium and high PA level. Demographic, socioeconomic, lifestyle factors, and comorbidities were assessed through questionnaires and registry data. Associations with PA levels were analyzed using multiple ordinal logistic regression. A total of 1,484 participants (32% women; median age 60.2 years) were included. Being in the oldest age group was significantly associated with lower odds of having a higher PA level (OR = 0.59; 95% CI [0.44, 0.77]), as well as female sex (OR = 0.73; 95% CI [0.59, 0.91]), regular/occasional smoking (OR = 0.37; 95% CI [0.26, 0.52]), and having one (OR = 0.61; 95% CI [0.48, 0.77]) or more comorbidities (OR = 0.45; 95% CI [0.30, 0.68]). A healthy diet was significantly associated with higher PA levels (OR = 1.84; 95% CI [1.46, 2.31]). Financial strain, level of education and alcohol consumption had no association with PA. Using data from a representative sample of middle-aged adults, the findings indicate that engagement in PA is primarily associated with lifestyle factors, demographic characteristics, and comorbidity. These identified characteristics may inform future research aimed at CVD populations with low PA. Further research should investigate mediating and moderating pathways to clarify how these factors influence PA.

SpyCEP dismantles neutrophil immunity via disorder-driven chemokine remodeling and GAG targeting

Streptococcus pyogenes (Group A Streptococcus ) employs sophisticated virulence strategies to evade human immunity, including secretion of the cell envelope protease SpyCEP, which cleaves and inactivates key neutrophil-attracting chemokines such as CXCL8. Here, we integrate cryo-electron microscopy, NMR spectroscopy, and native mass spectrometry to investigate how SpyCEP disrupts CXCL8 function. We demonstrate that a disordered aromatic and acidic region within the cleaved autocatalytic maturation loop (CAML) of SpyCEP mimics receptor N-domains and binds an allosteric site on CXCL8. The resulting interaction forms a dynamic fuzzy complex and is coupled to dimer dissociation, consistent with enhanced access to the cleavage site. This disorder-mediated substrate engagement differs from classical protease mechanisms that rely on rigid recognition interfaces. Additionally, glycan microarray and NMR analyses show that the CAML region mediates glycosaminoglycan (GAG) binding, suggesting a means for SpyCEP to maximize encounters with GAG-enriched CXCL8 reservoirs. Together, these findings provide a structural and biophysical framework for understanding how SpyCEP combines substrate engagement with GAG targeting to dismantle chemokine gradients and inhibit neutrophil recruitment. More broadly, this work highlights the role of intrinsic disorder in protease recognition and suggests avenues for anti-virulence therapies and vaccine strategies targeting SpyCEP.

Non-native plant integration into plant-insect pollinator networks in urban parks

Pollination sustains biodiversity and ecosystem functioning, yet it is increasingly threatened by biological invasions. Non-native plants frequently integrate into plant-pollinator networks, generating both positive and negative effects on native plants. However, most current knowledge derives from static or short-term studies of communities invaded by a few species, offering limited understanding of plant-pollinator dynamics in systems where many non-native species coexist under favorable environmental conditions. Here, we examined plant-insect pollinator networks in urban green areas of a Mediterranean city as a model system to evaluate the role of non-native plants in structuring plant-pollinator networks over the course of one year. We compared native and non-native plants in terms of (i) species-level parameters (normalized degree, strength, specialization, contribution to nestedness, and module roles), (ii) modularity and co-flowering patterns, (iii) persistence of interactions, and (iv) beta-diversity metrics. Non-native plants showed greater specialization and among-module connectivity, whereas native species exhibited greater species strength and within-module connectivity. Native–non-native pairs within modules co-flowered more often, suggesting simultaneous interactions that may foster both competition and facilitation. In contrast, non-native–non-native pairs displayed asynchronous flowering, potentially sustaining shared pollinators over longer periods and mitigating competition. Despite these differences, both groups contributed similarly to temporal network connectivity, and interaction turnover was unaffected by the proportion of non-native species. Overall, these findings reveal that while native and non-native plants play distinct roles in structuring plant-pollinator networks, they do not differ in their contribution to maintaining temporal connectivity. Our study underscores the importance of incorporating temporal and multispecies approaches when assessing the ecological consequences of biological invasions.

A DNA break-5mC cycle activates transposable elements in <i>Arabidopsis</i>

Plant genomic and epigenomic integrity are perpetually threatened by exogenous and endogenous DNA damage. However, the interplay between DNA damage, DNA methylation (5mC), and transposable element (TE) activity remains poorly understood. Here, we demonstrate that defective single-strand break (SSB) repair acts as a potent trigger for genome-wide TE derepression and extensive de novo DNA methylation in Arabidopsis . Mutations in ZDP/APE2, which encode conserved DNA 3’-end repair enzymes, impair the repair of 3’-blocked SSBs arising from base excision repair, ultimately leading to widespread TE activation. Concurrently, inefficient SSB repair activates the ATR–SOG1-mediated DNA damage response, which enhances the RNA-directed DNA methylation (RdDM) pathway to counteract TE activation by depositing 5mC. Paradoxically, the resulting methylation is excised by the DNA demethylase ROS1—a process that itself generates 3’-blocked SSBs requiring resolution by ZDP/APE2. In zdp ape2 mutants, ROS1-mediated 5mC excision produces additional SSBs, which in turn reactivate RdDM. This establishes a self-sustaining SSB–5mC cycle that perpetuates DNA damage and drives massive TE activation in the mutant. Our findings reveal a critical mechanistic link between SSB repair, DNA methylation dynamics, and TE derepression, positioning defective SSB repair as a major inducer of epigenomic instability.

Psychological assessment in patients suffering from chronic pain treated with Spinal Cord Stimulation: A prospective observational study protocol

Introduction Spinal cord stimulation (SCS) has been used to manage a variety of chronic pain conditions, including cancer population. People suffering from this medical condition often report strong limitations in their daily and work activities, and a deterioration of health-related quality of life. This study aims at investigating the relationship between psychological factors and intervention outcomes (pain reduction, removal of SCS). Further, the acceptability and satisfaction levels regarding the SCS intervention will be assessed. Methods and analysis Cancer survivors 1) suffering from chronic pain (VAS &gt; 4) from at least 6 months, 2) aged over 18 years, and 3) refractory to conventional pain therapies will be assessed at the pre-surgical visit (T0), and after 1, 2, 3, 5, and 8 months from T0. During each assessment, patients will undergo a clinical consultation on their ability to cope with chronic pain and fill out validated questionnaires on patients’ perception of pain (BPI, PCS), state and trait anxiety (STAI Y1-Y2), depression (BDI-II), and pain-specific resilience (PRS). Patients Global Impression of Change (PGIC) after SCS implantation will be also assessed. Data collection is ongoing. The correlation between continuous variables at considered time points will be evaluated with Pearson’s correlation coefficient. The difference in mean patients’ pain, anxiety, depression, catastrophizing tendency, and resilience according to the SCS removal request will be tested with Student’s t-test. Finally, the overall correlation between a psychological trait of interest and an intervention outcome over the entire course of observation will be estimated using a bivariate linear mixed effect model for longitudinal data. Ethics and dissemination The study protocol was reviewed and approved by the Ethics Committee of the European Institute of Oncology (Comitato Etico Territoriale Lombardia 2; R1865/23 – IEO 2028). The results of the study will be disseminated through a peer-reviewed journal. Registration details The study is registered on ClinicalTrials.gov (Identifier: NCT06761300).

Spontaneous phase separation and pattern formation in a lyotropic nematic mixture

Lyotropic liquid crystals can display rich phase behavior and self-organization, yet the physical principles underlying their self-assembly into large scale patterns remains understudied. Here, we combine theory, simulations, and experiments on Sunset Yellow-water chromonic mixtures to show that such materials spontaneously phase separate, even without assuming any underlying microscopic attraction between the molecular species. In our minimal model, demixing depends solely on the Onsager-like coupling between local nematogen density and orientational order. If such a coupling is sufficiently strong, nematic defects trigger the nucleation of isotropic droplets, which then coalesce due to elastic or interfacial tensions. We further show that strong anchoring of the director field at the interface arrests this coarsening process, resulting in a stable microphase-separated lamellar pattern. This self-assembled lamellar phase has striking and unusual features, including spontaneous undulations, heterogeneous layer spacing, long-lived glassy defect patterns and lamellar onions. Our results identify orientational-density coupling and elastocapillarity as fundamental mechanisms to guide self-assembly in lyotropic and chromonic liquid crystals.

Editorial Note: Apoptosis by [Pt(O,O′-acac)(γ-acac)(DMS)] requires PKC-δ mediated p53 activation in malignant pleural mesothelioma

Helfrich walls in nematic toroidal droplets in the presence of magnetic fields

We observe the formation of stable, magnetic field induced, alignment inversion walls, also called Helfrich walls, in nematic liquid crystals (NLCs) confined to a toroidal droplet with tangential anchoring at the boundaries. Walls are rare in NLCs due to their high energy compared to lower-dimensional defect structures. However, we find that Helfrich walls naturally form, transform, and migrate in this unconventional toroidal setting, generating intricate director-field configurations due to the interplay of confining geometry and external fields. Interestingly, the presence and number of point defects in the zero field nematic configuration of the toroid influences the types of walls that arise. In the absence of point defects, splay-bend walls emerge. By contrast, when a point defect pair exists on the toroidal surface, a wall not previously classified by Helfrich, and that involves both twist and splay-bend distortions, forms, and transforms. Our experiments and numerical simulations enable the quantification and elucidation of the physics behind the formation and transformation of these wall structures, extending the class of walls that were believed to occur in NLCs.

Sociocultural factors influencing alcohol use among Korean immigrant women: A scoping review

Background Although studies of alcohol use have explored the experiences of various ethnocultural communities in diverse countries such as Canada, there is a noticeable gap in primary research concerning the experiences of Korean immigrant women. With alcohol consumption rising among women relative to men in recent decades, a critical understanding of existing knowledge on alcohol use through the lenses of culture and gender is essential. Research Question What is the current state of evidence on alcohol use among Korean immigrant women? Methods This scoping review was conducted using the five-stage framework adapted from Arksey and O’Malley (2005) and extended by Levac et al. (2010), with reporting guided by the Joanna Briggs Institute PRISMA-ScR framework. Inclusion criteria comprised primary studies focused on both Korean immigrant women and alcohol use, from Canada, the United States, the United Kingdom, Australia, and New Zealand, restricted to adult ages and English-language publications. Inductive thematic analysis as outlined by Gottlieb et al. (2021) was employed to extract themes from the included articles. Results Seven articles met the inclusion criteria, all from the United States, revealing no existing research on this topic from any other eligible country. Three major sociocultural themes influencing alcohol use were identified: acculturation, religion, and social networks. Additional contextual factors including immigration-related stress and traditional Korean drinking customs were noted as plausible but less consistently reported influences. Conclusions Findings underscore the importance of adopting culturally sensitive, gender-specific approaches in public health initiatives targeting alcohol use among Korean immigrant women. Substantial gaps remain, particularly regarding the Canadian context.

Correction for Hong et al., Mechanistic links between coexistence, productivity, and stability in experimental grasslands

ShinyDataMatcher: A user-friendly application for integrating survey data

In this work, we introduce ShinyDataMatcher , a user-friendly R Shiny application designed to support the integration of survey data through statistical matching. The tool enables practitioners to import, explore and process survey data, harmonize variables, select appropriate matching variables, and apply a wide range of macro- and micro-level matching methods without writing any code. The application guides the user through the full workflow of a matching exercise, from data preparation to the creation of a synthetic matched dataset, and includes diagnostic tools for assessing matching quality. To illustrate its capabilities, we present an application based on the Italian Household Budget Survey (HBS) and the Survey on Household Income and Wealth (SHIW), where the goal is to fuse income and expenditure information and to construct a Social Accounting Matrix (SAM). The example also highlights how repeated random hot-deck imputations can be used to account for the additional uncertainty induced by statistical matching. Overall, ShinyDataMatcher provides a transparent and accessible environment for exploring, prototyping, and implementing statistical matching procedures, lowering the technical barriers that often limit their use in applied and official-statistics contexts.

Topological expansion of Boehm’s brushes via structured light

We report an entoptic phenomenon in which the classical two-lobed Boehm’s brushes are transformed into a multilobed structure by projecting spin–orbit-coupled light onto the human retina. These structured beams, composed of nonseparable superpositions of circular polarization and orbital angular momentum, produce azimuthally modulated entoptic patterns through polarization-dependent scattering in the retina. Unlike Haidinger’s brushes, which arise from dichroic absorption in the macula, the observed effect is driven by angular variations in scattering strength relative to the local polarization direction. In regions where scattering centers exhibit polarization orientations that converge toward a common point, their combined contributions reinforce one another, producing brighter and more sharply defined entoptic lobes whose number and orientation vary systematically with the topology of the spin–orbit stimulus. Psychophysical measurements across retinal eccentricities from 0.5 ° to 4 ° in eleven participants revealed that contrast detection thresholds decreased exponentially with eccentricity, consistent with polarization-sensitive scattering by isotropic structures in the nonfoveal retinal regions. From the psychophysical fits, the mean eccentricity at which the entoptic pattern reached a 50% threshold was r 50 = 1 . 03 ° with a 95% CI of [0.72, 1.34] ° , indicating that the spin–orbit-induced entoptic structure becomes perceptually robust at approximately 1 ° retinal eccentricity and that perception improves with increasing retinal eccentricity. Together, these findings demonstrate that spin–orbit light modulates scattering-based visual phenomena in previously unrecognized ways, enabling approaches for probing retinal structure and visual processing using topological features of light.

Inter-brain functional connectivity: Are we measuring the right thing?

Hyperscanning—the simultaneous recording of brain activity from multiple individuals—and the study of inter-brain synchronization is gaining popularity in social neuroscience. MEG/EEG hyperscanning studies often estimate inter-brain functional connectivity using phase-based metrics applied to oscillatory brain signals, assuming matching peak frequencies between the individuals studied. However, in reality peak frequencies typically differ between subjects and even between brain regions. Using simulated MEG/EEG signals, we systematically assessed how inter-individual frequency differences affect commonly used connectivity measures. Phase-based metrics were highly sensitive to frequency differences across individuals, whereas amplitude envelope correlation remained comparatively stable under these conditions. Our results underscore the need for connectivity metrics specifically tailored for inter-brain analyses. These findings are relevant to a range of disciplines that are increasingly integrating hyperscanning into their methodological toolkits.

IL-13 signaling in cDC2 is required for systemic anaphylactic responses

Cutaneous allergen sensitization (CAS) is a primary driver of atopic dermatitis (AD) and a key initiator of the “atopic march”, which can lead to systemic conditions such as food allergy and anaphylaxis. The type 2 cytokine interleukin-13 (IL-13) is an important regulator of high-affinity IgE antibodies, yet the precise cellular targets and mechanisms by which it orchestrates systemic allergic responses remain incompletely understood. Here, we evaluated the role of IL-13 in a murine CAS model that links skin inflammation to systemic anaphylaxis. Using cell-specific deletions of the IL-13 receptor α1 subunit ( Il13ra1 ), we identify conventional dendritic cells (cDCs), and not T or B cells, as the essential targets of IL-13 for generating high-affinity IgE. Single-cell transcriptomics reveal that IL-13 signaling acts specifically in a cDC2 subset characterized by high expression of CX3CR1, Clec10a (CD301a), and CD301b (Mgl2). Licensing by IL-13 endows these cDC2 with superior antigen-presenting capacity, characterized by the upregulation of MHC class II and costimulatory molecules, including CD301a, CD301b, and ICOSL. These mature cDC2s are mobilized from the periphery to the spleen by a CX3CR1-dependent mechanism, where they are uniquely equipped to induce the differentiation of IL-13-producing T follicular helper (T FH 13) cells. This cascade results in robust germinal center reactions and production of pathogenic, high-affinity IgE. Our findings define an IL-13–cDC2 axis that functions as a critical regulator of the atopic march, providing a mechanistic rationale for the clinical efficacy of IL-13-targeted therapies in allergic diseases.

Temporal binding and social perception – A replication study

Previous studies reported action-effect binding for social contexts in a variety of experimental contexts. When confronted with social information, perceptual or conceptual, participants underestimated the duration of time intervals between their actions and subsequent social events, as compared to only observing social events. Current evidence demonstrates stronger temporal binding both for conditions of believing in an interaction with a social partner, and for perceiving an interaction partner. Fewer data exist about the influence of exclusively perceptual social cues such as interaction with a face stimulus. We here report on a study investigating this so-called social hyperbinding for face perception. 40 participants performed a duration estimation task in four blocks involving either the active manipulation of gaze direction, the active manipulation of a geometric pattern, the passive observation of gaze direction, or the passive observation of movement in a geometric pattern. Although results indicated trends towards shorter time estimates for the active manipulation of gaze, with no visible differences in time perception for the other three conditions, our analysis did neither confirm temporal binding nor social hyperbinding for face stimuli. This finding challenges the theory of social hyperbinding for face stimuli. While stronger temporal binding for real and believed interactions with another human has been reliably replicated in the literature, the results from this experiment cannot confirm increased binding for face-like stimuli.

Spatially structured heterogeneity shapes large-scale cortical dynamics in a model of the human cortex

The human brain displays substantial spatial variability in molecular, anatomical, and physiological organization. Yet, how this heterogeneity shapes large-scale neuronal dynamics remains poorly understood. To address this question, we employed a biologically informed large-scale cortical model capable of generating distinct brain states, from awake-like to sleep-like dynamics. Our model was constrained by empirical human structural connectivity (SC) and regional cholinergic muscarinic receptor (CHRM) maps derived from transcriptomic data, together with complementary positron emission tomography (PET)-based receptor maps. These regional maps were implemented as modulators of adaptation-related excitability. We found that modulating excitability according to the spatial organization of CHRM maps significantly impacted large-scale cortical dynamics: It not only facilitated network synchronization but also enhanced information flow between cortical regions. Importantly, these effects were conserved across transcriptomic and PET-derived maps and could not be fully reproduced by multiple null models preserving generic forms of heterogeneity. Moreover, we addressed a particularly intricate dynamic regime characterized by the coexistence of localized sleep-like activity within otherwise awake-like states. We showed that the emergence of these sleep-like slow waves was a byproduct of both regional levels of neuronal adaptation and SC. In summary, our findings highlight the critical role of molecular and anatomical heterogeneity in shaping widespread cortical dynamics, suggesting broad avenues for linking microscale diversity to macroscale function.

A human 3D BBB chip model of acute stroke simulating a reversible penumbra

Ischemic stroke is a leading cause of mortality and disability worldwide. However, existing models often fail to replicate key aspects of human pathophysiology, particularly blood–brain barrier (BBB) dysfunction and the salvageable ischemic penumbra. We developed a three-dimensional BBB chip model of acute ischemic stroke that reproduces penumbra-like, partially reversible BBB injury. This platform integrates a microfluidic BBB chip (Emulate) with parallel Transwell inserts to facilitate complementary structural, molecular, and functional analyses. Ischemia-like injury was induced using 2.5 μM antimycin A for 1 hour under oxygen-glucose deprivation conditions, followed by medium replacement to simulate reperfusion. Therapeutic hypothermia (33°C) was also applied during the reperfusion phase. The combination of reperfusion and hypothermia resulted in the most pronounced restoration of BBB integrity compared with reperfusion alone. The Emulate chip enabled structural evaluation of endothelial morphology, while the transwell model showed concordant recovery of BBB-related markers, including ZO-1 and VE-cadherin, along with decreased expression of the hypoxia-associated marker HIF-1α. This integrated platform enabled evaluation of BBB injury and recovery under ischemia- and reperfusion-like conditions. Our human cell-based 3D BBB stroke model captures key BBB-related features of penumbra-like injury and provides a human-relevant in vitro platform for investigating stroke pathophysiology and evaluating therapeutic strategies.