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Zhu, J.

Publications and source records attributed to Zhu, J..

At least 37 records · Page 2Linked to original sources

Dynamic architecture of the Rixosome reveals mechanism of activation and ITS2 processing

Eukaryotic ribosome assembly requires the coordinated processing and extensive remodeling of pre-rRNAs. During late nuclear maturation of the 60S subunit, sequential removal of the internal transcribed spacer 2 (ITS2) is initiated by endonucleolytic cleavage at site C2 by the conserved Las1 nuclease. Las1 acts together with the kinase Grc3 and the Rix1 complex to form the Rixosome, which also functions in transcriptional regulation. However, the assembly of the Rixosome, its recruitment to pre-ribosomes, and its activation for ITS2 cleavage remain unclear. Here, we present cryo-EM structures of the human LAS1 complex, two structures of the isolated Rixosome and nine transition states of Rix1-bound pre-60S particles from Schizosaccharomyces pombe. These structures reveal a dynamic Rixosome architecture in which the heterotetrameric Las1 complex engages one or two copies of the Rix1 complex. Rix1 binding is highly flexible in the human Rixosome but rigid in the yeast complex. The isolated yeast Rixosome remains inactive, but binding to the pre-60S particle triggers a structural rearrangement that allows for substrate engagement and activation of the nuclease. Together, our results define the dynamic architecture of the Rixosome and provide a structural framework for ITS2 processing during nuclear maturation of the eukaryotic 60S ribosomal subunit.

biochemistry↗

A Distal Super-Enhancer Regulate CD47 Expression and Promotes Macrophage-Mediated Phagocytosis of Ovarian Cancer Cells

CD47 is an innate immune checkpoint that can protect tumor cells from macrophage-mediated clearance, but the regulatory mechanisms underlying its expression in ovarian cancer (OC) remain incompletely understood. Here, we investigated the epigenetic regulation of CD47 expression in OC. Single-cell and spatial transcriptomic analyses indicated preferential CD47 expression in malignant epithelial cells. Integration of publicly available epigenomic datasets revealed a distal regulatory region with enhancer-associated features across ovarian cancer models and HGSOC patient samples. A genome-wide CRISPR-Cas9 screen further implicated PITX2 and EP300 as regulators of CD47 expression. Chromatin immunoprecipitation assays showed PITX2 and EP300 occupancy at candidate enhancer elements within this CD47 regulatory region. Comparative epigenomic analyses suggested that features of this regulatory landscape are also present in fallopian tube epithelial cells and become more prominent in OC. Together, these findings support a PITX2-EP300 regulatory axis associated with CD47 expression and provide evidence for its potential contribution to tumor immune evasion.

cancer biology↗

NAE1-Dependent Protein Neddylation Preserves Endothelial Identity and Vascular Integrity

Background: Endothelial dysfunction is a central driver of cardiovascular and inflammatory diseases, yet the post-translational mechanisms that preserve endothelial homeostasis remain incompletely understood. Protein neddylation, the covalent conjugation of a ubiquitin-like modifier, regulates diverse cellular processes, yet its physiological role in the vascular endothelium remains unknown. This study investigated whether protein neddylation is required to preserve endothelial identity and vascular homeostasis. Methods: We generated tamoxifen-inducible endothelial-specific Nae1 knockout mice to inhibit neddylation and combined bulk RNA sequencing, single-cell and single-nucleus transcriptomics, quantitative proteomics, biochemical analyses, and gain- and loss-of-function approaches to define the role of endothelial neddylation in vascular homeostasis and inflammatory injury. Results: Endothelial-specific Nae1 deletion caused rapid mortality associated with vascular leakage, platelet accumulation, inflammation, and multi-organ injury. Multi-omics analyses demonstrated profound loss of endothelial identity, characterized by suppression of core endothelial programs and activation of inflammatory, procoagulant, and pyroptotic pathways. Single-cell analyses revealed progressive endothelial dysfunction culminating in depletion of the endothelial population and remodeling of the vascular niche. Mechanistically, endothelial neddylation deficiency activated gasdermin D (GSDMD)- and gasdermin E (GSDME)-dependent pyroptosis, whereas dual inhibition of GSDMD and GSDME markedly attenuated inflammatory transcriptomic remodeling, vascular injury, hepatocyte death, immune cell infiltration, and platelet accumulation. Translational analyses demonstrated reduced endothelial neddylation in experimental endotoxemia and decreased expression of neddylation pathway components in human atherosclerosis and COVID-19 datasets. Conversely, restoration of endothelial neddylation partially reversed inflammatory endothelial transcriptomic reprogramming in vivo. Conclusions: NAE1-dependent protein neddylation is an essential regulator of endothelial identity and vascular integrity. Loss of endothelial neddylation promotes gasdermin-dependent pyroptosis and thrombo-inflammatory vascular injury, whereas restoration of the neddylation pathway mitigates inflammatory endothelial dysfunction. These findings identify endothelial neddylation as a fundamental mechanism maintaining vascular homeostasis and a potential therapeutic target for cardiovascular and inflammatory diseases.

pathology↗

Extracellular water withdrawal drives disease resistance in the phyllosphere

A central question in immunity is how hosts arrest pathogen growth. Diverse plant pathogens create a water-soaked niche in host tissue essential for pathogenesis, yet how water shapes infection outcome is unknown. Using genetics and hyperspectral imaging, we show that extracellular water status is rate-limiting for both compatible and incompatible interactions. We find that the hypersensitive response of effector-triggered immunity (ETI) is, mechanistically, a desiccation event. Water loss imposes osmotic stress that arrests bacterial division while the pathogen remains alive and metabolically active, rather than killing it. Restoring apoplastic water reverses this stasis and licenses growth despite intact immune signaling and cell death. Water status, not immune signaling per se, gates pathogen growth. This reframes ETI as a controlled desiccation mechanism and identifies hydration as a decisive lever on disease outcome.

plant biology↗

Design and Assembly of Combinatorial DNA Barcodes for Probe-based Genomics Applications

Probe-based genomics technologies are extending molecular analysis into intact tissues and fixed cells, yet strategies to decode complex experimental conditions encoded in cellular RNA remain limited. Here we present a modular framework that integrates custom software tools with purpose-built cloning reagents to design, assemble, validate, and deploy combinatorial DNA barcodes. Combinatorial barcodes comprise spatially adjacent collections of known sequences, enabling millions of unique molecules to be efficiently distinguished using a limited set of probes. Our software tools integrate with optimized assembly plasmids and whole plasmid long-read sequencing for high-fidelity construction and structural validation of diverse combinatorial barcode architectures. Assembled barcode libraries are flexibly transferred into user-modified expression vectors to support diverse downstream experimental applications. We showcase the versatility of this framework by assembling two structurally distinct combinatorial barcode libraries, each containing millions of unique sequences. Following rabies virus-based delivery to the mouse brain, we validate in vivo decoding of a combinatorial barcode architecture capable of distinguishing ~16.3 million expressed RNAs through probe-based in situ sequencing. Our framework for flexible and accurate combinatorial barcode construction fills a technically demanding niche delivering cost-effective molecular reagents for multiplexed experimentation on current and evolving probe-based genomics platforms.

genomics↗

The transcription factor Bhlhe40 promotes inflammatory functions of ILC2s through inducing GM-CSF while inhibiting IL-10 expression

Group 2 innate lymphoid cells (ILC2s) regulate type 2 immune responses partly by recruiting eosinophils, but the mechanisms underlying this process remain incompletely understood. Although both ILC2s and type 2 T helper (Th2) cells are capable of expressing IL-5, a cytokine critical for recruiting eosinophils, ILC2s are more potent than Th2 cells in this process. Here, we show that the transcription factor Bhlhe40 promotes GM-CSF production in ILC2s, and Bhlhe40 expression in ILC2s is required for efficient eosinophil recruitment during both papain- and helminth-induced type 2 immune responses. However, Bhlhe40 deficiency in ILC2s had no effect on the production of classical type 2 cytokines, including IL-4, IL-5 and IL-13, despite Bhlhe40 being required for type 2 cytokine production by Th2 cells. Furthermore, in contrast to ILC2s, Th2 cells produced little GM-CSF and administration of GM-CSF rescued eosinophil recruitment in ILC2-specific Bhlhe40-deficient mice. In addition to promoting GM-CSF expression, Bhlhe40 repressed IL-10 production in ILC2s as it did in Th2 cells, particularly during chronic inflammation. Single-cell transcriptomic analyses further supported this regulatory network, and the ChIP-Seq data revealed direct binding of Bhlhe40 to the enhancer and promoter regions within the Il10 and Csf2 loci. Collectively, our findings show that Bhlhe40 exerts distinct gene regulatory functions in ILC2s and Th2 cells, and Bhlhe40 modulates the inflammatory and anti-inflammatory functions of ILC2s by promoting GM-CSF while suppressing IL-10 production.

immunology↗

TRAPPC8 Is an Endogenous Brake on UFMylation That Suppresses Tauopathy

UFMylation, a ubiquitin-like protein modification, drives tau spread through the brain, but what keeps this process in check has remained unclear. We show that TRAPPC8 acts as a natural brake on UFMylation, binding directly to the E1 enzyme UBA5 to dampen pathway activity. In Alzheimer's disease brain tissue, TRAPPC8 is reduced while UFMylation is elevated, suggesting this brake fails as disease progresses. Sustaining UFMylation in human iPSC-derived neurons increased tau aggregation and spread while disrupting lysosomal function and lipid balance. Restoring TRAPPC8-UBA5 binding reversed these defects through the lysosomal protein CLN8, which restored lysosomal function and reduced tau pathology. Notably, expressing just the UBA5-binding region of TRAPPC8 was enough to suppress tau pathology in vivo, marking it as a promising therapeutic target.

neuroscience↗

Structure-aware deep learning predicts influenza antigenicity and guides vaccine strain recommendation

The continuous accumulation of genetic mutations in influenza A viruses (IAVs) drives antigenic drift, necessitating precise antigenic prediction for optimal vaccine strain selection. While sequence-based methods have advanced antigenic surveillance, they neglect the three-dimensional structural context that fundamentally dictates viral antigenicity. Here, we introduce Vir3D, which leverages ESMFold-derived structural information from amino acid sequences to precisely predict viral antigenicity and guide vaccine strain selection. Across both human H3 and highly pathogenic avian H5 subtypes, Vir3D not only accurately discriminates antigenic variants and infers pairwise antigenic distances, but also mechanistically delineates key structural residues driving viral immune evasion. In a decade-long retrospective analysis, Vir3D-prioritized vaccines consistently achieve broader antigenic coverage of circulating strains than World Health Organization (WHO) recommendations. Crucially, Vir3D successfully predicts that the emerging U.S. dairy cattle H5N1 virus (TX/24) remains antigenically stable relative to clade 2.3.4.4b vaccine strains, and subsequent wet-laboratory validation of hemagglutination inhibition (HI) assays definitively corroborates this finding. Overall, Vir3D establishes a powerful, structure-driven framework for proactive influenza surveillance and pandemic preparedness.

bioinformatics↗

Collaborative multi-agent intelligence uncovers subtype-selective allosteric sites at GPCR-lipid interaction interface

Closely related G protein-coupled receptor (GPCR) subtypes often share highly conserved orthosteric pockets, making subtype-selective ligand development challenging. Here, we developed a five-agent workflow to systematically identify divergent protein-membrane-interface sites across class A GPCRs and exploit them for selective allosteric ligand discovery. By combining dMaSIF-derived surface fingerprints with Ballesteros-Weinstein (BW) position alignment, we compared structurally equivalent membrane-facing regions and identified the three most divergent hotspots for each of 163 receptor pairs. These regions showed substantial spatial overlap with experimentally characterized allosteric sites. Paired target-off-target screening of one million lead-like compounds, followed by detail-mode redocking and multi-seed consistency filtering, yielded 352 receptor-pair-specific candidates corresponding to 344 unique compounds across 104 receptor pairs. These candidates, together with their divergent sites and predicted selectivity profiles, were integrated into a searchable database. Our findings establish a scalable strategy for translating GPCR membrane-interface divergence into precise allosteric sites and testable subtype-selective ligand candidates.

biochemistry↗

Individual-tree phenology reveals climate-dependent responses to urban thermal heterogeneity across U.S. cities

Rapid urbanization is altering the seasonal life cycles, or phenology, of urban trees. Diverse species composition and heterogeneous urban thermal environments can generate pronounced phenological variation within cities, yet such fine-scale seasonality remains poorly understood. This limits the ability to anticipate localized ecosystem services and disservices, including canopy cooling and pollen exposure. Here, we analyzed individual-tree phenology and their response to temperature across 76 cities in the contiguous United States by integrating municipal street-tree inventories, PlanetScope satellite time series, and 1-km near-surface urban air-temperature data. We quantified intra-city and cross-city phenological variation, estimated species-level associations between phenology and fine-scale urban temperature, and tested whether these associations vary with regional climatic context. Intra-city phenological variation rivaled cross-city variation for 33.3% of species in spring and 53.7% in fall. Temperature-phenology associations were widespread: warmer local urban environments were associated with earlier spring onset in 51.5% of cases and delayed fall senescence in 66.5%, with approximately two-thirds of these responses cascading to longer growing seasons. These associations varied with regional climate, generally showing stronger spring advancement and fall delay in cooler and wetter cities, but weaker or reversed responses in hotter or drier cities. Together, these results suggest that urban tree phenology is shaped by interactions among fine-scale urban temperature, species identity, and regional climate. Accounting for this heterogeneity can improve predictions of urban forest function and inform climate-resilient species selection and management. Plain Language SummaryStreet trees within the same city do not all leaf out in spring or lose their leaves in fall at the same time. This seasonal timing, called phenology, affects services such as shade and cooling, as well as disservices such as pollen exposure. We studied individual tree phenology and their temperature response in 76 U.S. cities using street-tree records, satellite observations, and local air-temperature data. We asked how much phenology varies within cities, how it responds to neighborhood-scale temperature differences, and whether these responses depend on regional climate. Variation within a city was often as large as variation among cities, especially in fall. Trees in warmer parts of cities generally leafed out earlier and lost their leaves later, often extending the growing season. However, these responses were stronger in cooler and wetter cities and weaker or sometimes reversed in hotter or drier cities. These findings show that urban tree seasonality depends on local temperature, species identity, and regional climate, which should be considered when selecting and managing trees for future cities. Key PointsO_LISubstantial within-city phenological variation is not captured by city-level averages. C_LIO_LIFine-scale temperature shaped phenology and growing-season length, but responses varied across species, cities, and phases. C_LIO_LITemperature sensitivity depended on regional climate, weakening or reversing in hotter or drier cities. C_LI

ecology↗

Evolution-inspired multi-objective Bayesian optimization for protein engineering

Protein engineering requires efficient navigation of vast sequence spaces under limited evaluation budgets, especially when multiple properties must be optimized simultaneously. We developed Evolution-inspired Multi-Objective Bayesian Optimization (EvoMOBO), an active-learning framework that integrates path-dependent sequence generation, global competition among generated variants, and explicit multi-objective optimization. Benchmarking against state-of-the-art methods on complete steroid receptor DNA-binding domain and ParD3 antitoxin landscapes demonstrated robust target-region enrichment, Pareto-front advancement, and sequence diversity across two- and three-objective tasks. In the DBD landscape, simulation-derived geometric descriptors served as labels for both initialization and iterative updating, enriching variants with favorable measured activities without experimental labels. Building on this validation, we applied EvoMOBO to two enzyme-engineering tasks using simulation-derived mechanistic descriptors, with experiments reserved for final validation. For an old yellow enzyme (GkOYE), 16 of 26 tested variants outperformed the wild type, and the best increased non-native oxidative dehydrogenation conversion from 17.5% to 95%. For a formate oxidase (AoFOx), EvoMOBO identified aggregation-resistant variants, two of which nearly doubled diethyl phthalate degradation in a photoenzymatic cascade. Together, these results establish EvoMOBO as a modular framework for multi-objective protein engineering using experimental or mechanism-derived labels.

bioengineering↗

Coronary Artery Disease Transcriptomics Reveals Two Drivers of the Endothelial Cell SR-BI Expression and LDL Transport that Underlie Atherosclerosis

Atherosclerosis is initiated by circulating low-density lipoprotein (LDL) cholesterol transfer into the artery wall, which is mediated by scavenger receptor class B, type I (SR-BI) in endothelial cells(1). Employing single-cell RNA sequencing in human coronary artery disease (CAD) samples, here we show that endothelial SR-BI expression is increased in atheroma, and in endothelial cells with a transcript signature indicative of responding to disturbed blood flow. In vivo in mice hypercholesterolemia and disturbed blood flow independently upregulate endothelial SR-BI; the flow-related upregulation initiates endothelial cell LDL uptake and atherogenesis. Guided by transcription factor networks, it is revealed that HIF-1 binding to human Scarb1 Intron 1 governs endothelial SR-BI transcription, and in mice HIF-1 drives hypercholesterolemia-related SR-BI upregulation and artery LDL uptake. Thus, the two major instigators of atherosclerotic lesion formation, hypercholesterolemia and disturbed blood flow, both upregulate endothelial SR-BI to drive the LDL transport that underlies the disorder. Targeting the processes regulating endothelial SR-BI potentially represents a new therapeutic strategy against CAD.

cell biology↗

Parallel Emergence and Adaptive Evolution of Multicountry Circulating Mycobacterium abscessus Clones

Mycobacterium abscessus (MAB) is a recently emerged bacterial pathogen causing an increasing number of human infections. Despite increasing recognition that different circulating clones contribute disproportionately to MAB population expansion, the global population dynamics and the evolutionary pressures underlying the emergence and spread of MAB clones remain incompletely understood. We analyzed 11,314 publicly available genomes of MAB isolates sampled from 30 countries and identified 38 recently emerged clones that have spread across multiple countries, including the seven previously described Dominant Circulating Clones (DCCs). There was a significant association between the time since the emergence of each circulating clone and its population size, with older clones generally having larger populations. Among the 26 circulating clones of M. abscessus subsp. abscessus, 12 carried the macrolide-susceptible erm(41) 28C genotype, highlighting clone-level variation in macrolide susceptibility. Genome-wide analyses identified 55 genes that were previously under purifying selection but showed evidence of positive selection during recent clade expansion. Collectively, our findings reveal that the recent global expansion of MAB comprises a broad continuum of newly emerged clones, accompanied by widespread shifts in gene-level selective pressures indicative of ongoing adaptation to new ecological environments.

microbiology↗

Integrative spatial profiling of 3D genome organization and gene expression in tissue

The interplay between 3D genome architecture and transcriptional activity is fundamental to gene regulation. However, existing methodologies cannot simultaneously measure these modalities within intact tissues, limiting our understanding of how genome organization coordinates transcriptional programs across diverse cell types and spatial microenvironments. Here, we introduce Spatial Hi-C-RNA, a spatial multi-omics technology that enables the genome-wide co-mapping of chromatin conformation and transcriptome directly from the same tissue section at near- single-cell resolution. Applied to the mouse embryo and adult brains, Spatial Hi-C-RNA generated high-resolution tissue maps revealing that chromatin organization and gene expression jointly define spatially coherent domains aligned with histological structures. While concordant features were observed across modalities, distinct domain patterns also emerged, indicating that chromatin structure and transcription each contribute complementary layers of spatial regulation. We further demonstrated the robustness and biological insight of Spatial Hi-C-RNA in human melanoma, where both modalities delineated tumor boundaries and microenvironmental niches. Notably, chromatin maps revealed fine-scale tumor subdomains undetectable by transcriptomic profiling alone, highlighting the added resolution provided by spatial chromatin architecture. Integrated analysis revealed that multiscale 3D genome features, from A/B compartments and topologically associating domains to chromatin loops, are closely coupled with domain- and cell-type-specific transcriptional programs. In addition, Spatial Hi-C-RNA resolves spatiotemporal dynamics underlying embryonic lineage specification and tumor progression. Together, these capabilities extend the spatial omics landscape beyond transcriptome and epigenome profiling to the level of chromatin organization, establishing an integrative framework for understanding tissue biology across development and disease.

genomics↗

Multiplex genome engineering in yeast using the TIGR-Tas system

Tandem Interspaced Guide RNA (TIGR)-TIGR-associated (Tas) systems are a newly discovered family of ultracompact, modular RNA-guided DNA-targeting proteins that function without a protospacer adjacent motif (PAM) requirement. Their utility as genome engineering tools in microbes remains unexplored. Here, we report the first functional implementation of TIGR-Tas in Saccharomyces cerevisiae for genome engineering. We show that TasR from Parcubacteria (ParTasR) can be programmed by user-defined tigRNAs to generate targeted DNA double-strand breaks at yeast endogenous loci. By co-delivering ParTasR with customized tigRNAs and donor templates, we achieved precise gene fragment deletion and targeted codon substitutions at multiple genomic loci. The multiplex genome engineering capability of this TIGR-Tas system was demonstrated through high-efficiency multiplex gene disruption and chromosomal assembly of a lycopene biosynthesis pathway while inactivating an endogenous gene. This work establishes TIGR-Tas as a valuable addition to the yeast genome engineering toolbox, particularly for applications requiring PAM-independent targeting or compact delivery.

synthetic biology↗

Structural homology reveals cerato-platanins as conserved antimicrobials repeatedly co-opted for fungal host colonization

Fungal cerato-platanins (CPs) are small ([~]12 kDa) secreted proteins broadly conserved across filamentous fungi and implicated in a striking diversity of biological processes, ranging from fungal development to interactions with plant hosts. However, a core molecular activity unifying these diverse functions has remained elusive. Fungal pathogens secrete effector proteins, including CPs, to manipulate host physiology and promote colonization. Increasing evidence demonstrates that particular fungal effectors possess antimicrobial activity that enables pathogens to reshape host-associated microbiota during infection, and that antimicrobial activity is strongly enriched among evolutionarily conserved secreted proteins. These findings suggests that intermicrobial competition represents an ancient and fundamental fungal trait, and that several effectors that manipulate plant hosts evolved from ancient antimicrobials. Here, we identify antimicrobial activity as the conserved core function of the CP family, from which diverse roles in environmental adaptation and host interaction evolved. Structural analysis of the Verticillium dahliae secretome identified a structural cluster containing the CP protein CP1 and the previously characterized antimicrobial effector Ave1, providing the first structural link between CPs and antimicrobial activity. Accordingly, functional assays demonstrate that V. dahliae CPs exhibit selective antimicrobial activity in vitro. Expanding this analysis to a wide diversity of phylogenetic lineages and ecological lifestyles revealed that antimicrobial features are broadly conserved across the CP family. Together, our findings identify antimicrobial activity as an ancient and conserved molecular function that unifies the CP family and support a model in which host-manipulating effector functions evolved from ancestral proteins that mediate inter-microbial competition.

microbiology↗

Structured Sparsification of Signal-Transmission Networks Enhances Visual Information Coding

The brain can rapidly adjust sensory processing according to behavioral context from moment to moment without altering its underlying anatomical wiring. Such flexibility is thought to arise from dynamic reconfiguration of the effective network through which signals propagate, yet the principles governing such reconfiguration and its consequences for coding remain unclear. Here, we exploited the distinct stationary and locomotion states within the same recording sessions and addressed this question by inferring directed, millisecond-scale signal-transmission networks at single-neuron resolution in behaving mice. Locomotion was accompanied by a counterintuitive structured sparsification of the inferred network: interactions became fewer but more temporally precise, while the remaining interactions were more local, modular, feature-specific, and feedforward. We then used theoretical analysis and controlled perturbations of multi-area rate models to systematically determine how each empirically observed component of this reorganization affects population coding. We found that sparsification reduced shared variability; local organization reduced signal-noise alignment; feature-specific interactions sharpened selectivity; and a more feedforward architecture accelerated decoding. These results provide an experimentally grounded mechanism by which behavioral state reorganizes neuronal interactions given the same sensory inputs, and suggest structured sparsification as an underlying principle of network reconfiguration that supports accurate and faster sensory coding.

neuroscience↗

Diaphragmatic EMG Uncovers Left-Right Diaphragmatic Imbalance After Ischemic Stroke with Implications for Motor Recovery

BackgroundStroke-associated respiratory dysfunction has been increasingly recognized, yet whether diaphragmatic impairment after stroke involves lateralized neuromuscular imbalance remains unclear. MethodsIn this controlled experimental study, transient left middle cerebral artery occlusion and reperfusion (tMCAO) was performed in mice, followed by multimodal assessment of treadmill-based peak oxygen uptake testing, diaphragm ultrasonography, bilateral diaphragmatic electromyography (dEMG) with simultaneous respiratory flow monitoring, behavioral assessment, and whole-mount immunofluorescence imaging of the diaphragm. ResultsIschemic stroke reduced exercise capacity and diaphragmatic excursion without detectable diaphragm thinning, indicating functional impairment rather than overt atrophy. Bilateral dEMG revealed a subacute left-right asymmetry in inspiratory activation, with the ipsilesional hemidiaphragm exhibiting reduced amplitude, decreased area under the curve, and altered burst duration, consistent with a relative contralateral-dominant pattern rather than frank hyperactivation. Whole-mount imaging demonstrated asymmetric nerve remodeling, characterized by a nadir in ipsilesional nerve fiber density at 1 week and partial recovery by 2 weeks after stroke. These structural changes closely paralleled the dEMG alterations, suggesting a neural substrate for lateralized dysfunction. Exploratory analyses further linked dEMG parameters with motor recovery. ConclusionsThese findings provide novel evidence that ischemic stroke induces a left-right diaphragmatic neuromuscular imbalance, with structural and functional correlates in diaphragmatic innervation. The study further supports the utility of bilateral dEMG as a functional marker for assessing diaphragmatic dysfunction and monitoring recovery after stroke. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=190 SRC="FIGDIR/small/737872v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@10001a4org.highwire.dtl.DTLVardef@16bc3d5org.highwire.dtl.DTLVardef@5e40aorg.highwire.dtl.DTLVardef@be53b7_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗