Biology subjects

Li, J.

Publications and source records attributed to Li, J..

At least 199 records · Page 11Linked to original sources

bioRxiv2026

Insights into Genome Ejection by a Therapeutic phiKMV-like Bacteriophage

Ar-KM is a phiKMV-like therapeutic bacteriophage used in clinical candidate phage therapy cocktails to treat lung infections caused by Pseudomonas aeruginosa. Here, we present an integrative structural atlas of Ar-KM proteins using cryo-EM, proteomics, and bioinformatics. From a single purified Ar-KM preparation, we identified three distinct populations: mature DNA-filled virions, open-nozzle particles with ejection proteins extending from the tail, and closed-nozzle empty particles. Near-atomic-resolution reconstructions of all three states enabled us to build atomic models for eleven structural proteins. The mature virion revealed the pre-ejection conformation of three ejection proteins, gp41, gp42, and gp43, homologous to coliphage T7s gp14, gp15, and gp16, respectively. Unlike T7, peptidoglycan hydrolase activity associated with the ejectosome resides in the gp15-like periplasmic tunnel protein gp42, whereas in T7 the lysozyme-like domain is located at the N-terminus of gp16, underscoring the structural plasticity and evolutionary mosaicity of ejection proteins. We further identified a short -helical factor, gp34, present in eight copies at the mismatched interface between the portal barrel and gp41. Gp34 forms a cage within the nozzle, acting as a molecular wedge that stabilizes the open conformation and permits gp41 to assemble into a hexameric channel during ejection. Evolutionarily, gp34 appears to be an ortholog of the essential gene gp7.3 in phage T7 and is conserved across sequenced phiKMV-like phages. We propose that this protein functions as an ejection protein assembly factor, stabilizing the open nozzle during infection and allowing the coordinated exit of ejection proteins and their assembly into a DNA-ejectosome. HighlightsO_LIStructural polymorphism of phage Ar-KM defines open and closed nozzle states C_LIO_LIEjection proteins gp43, gp42, and gp41 assemble in a 4:8:8 stoichiometry C_LIO_LILysozyme domain shows modular positioning within the ejectosome C_LIO_LIPortal protein acts as the primary barrier to genome leakage C_LIO_LIWedge protein gp34 (T7 gp7.3 ortholog) bridges portal and gp41 C_LI

biophysics↗

bioRxiv2026

Identification of Reductases Catalyzing Benzyl Alcohol Formation during Salicylic Acid Biosynthesis in Plants

Salicylic acid (SA), a central hormone in plant immunity, is biosynthesized via a recently elucidated phenylalanine-derived pathway in most seed plants. This pathway requires benzyl alcohol as a key substrate for the formation of the SA precursor benzyl benzoate. However, how benzyl alcohol is produced in plants was unclear. Here, we identify a two-step conversion of benzoyl-CoA to benzyl alcohol via benzaldehyde in Nicotiana (N.) benthamiana. From a forward genetic screen for SA-deficient mutants, the and {beta} subunits of heterodimeric benzaldehyde synthase (BalS) involved in the conversion of benzoyl-CoA to benzaldehyde were found to be required for SA biosynthesis in N. benthamiana. Further reverse genetic analysis revealed that the NADPH-dependent benzaldehyde reductase (BalR1) acts downstream of BalS to convert benzaldehyde to benzyl alcohol. Interestingly, OsBalR1, but not OsBalS or OsBalS{beta}, is required for maintaining high basal SA levels in rice, suggesting the presence of redundant benzoyl-CoA-reducing activities or alternative biosynthesis routes for benzyl alcohol production. Together, this work defines the missing enzymatic steps in phenylalanine-derived SA pathway and provides insights into the evolutionary diversification of SA production strategies in plants.

plant biology↗

bioRxiv2026

cfMIND: A read-level methylation framework for accurate non- invasive disease detection using cell-free DNA

Plasma cell-free DNA (cfDNA) emerged as a promising non-invasive biomarker for cancers. However, reliable detection remains challenging due to the low abundance of tumor-derived cfDNA fragments and the dilution of informative methylation signals when aggregated into region-level features. Here, we propose a novel approach cfMIND, an efficient and robust machine-learning framework that leverages stratified read-level methylation signals to preserve rare cell-type-specific information and enhance detection sensitivity. By avoiding information loss inherent to conventional aggregation strategies, cfMIND enables more accurate and stable inference across diverse conditions. cfMIND is compatible with various cfDNA methylation sequencing technologies and cancer types. Across multiple cancer datasets (n = 868), cfMIND achieves high performance (AUROC up to 0.966) and maintains strong accuracy even at ultra-low sequencing depth (0.2x) and in early-stage cancers. Notably, cfMIND demonstrates exceptional robustness, generalizing effectively across cohorts and platforms without the need for model retraining. These results highlight its potential utility in heterogeneous experimental and clinically relevant settings. Beyond cancer detection, cfMIND is readily extendable to non-malignant diseases, as demonstrated by its ability to capture disease-associated methylation alterations in amyotrophic lateral sclerosis (ALS). Functional investigations on cfMIND-identified features further reveal enrichment in key regulatory regions implicated in disease pathogenesis and recapitulate tissue- and single-cell-level methylation and transcriptional programs underlying tumor biology. Collectively, cfMIND represents a significant advancement in the field, offering a broadly applicable, functionally interpretable, and high-resolution framework for non-invasive disease detection.

bioinformatics↗

bioRxiv2026

S2F-agent: Skill-grounded agent for Sequence-to-Function computational genomics workflows

Sequence-to-function (S2F) models offer a revolutionary paradigm for genotype-phenotype mapping, yet their broader application is bottlenecked by the need for reliable orchestration and interpretation across a fragmented model ecosystem. While general-purpose language models can automate scientific workflows, they are not inherently grounded in the model-specific execution constraints required for robust S2F analysis. Here, we present S2F-Agent, a human-in-the-loop framework designed for the verifiable orchestration of the heterogeneous S2F ecosystems. The framework employs a contract-based harness to bridge model-specific capabilities (Skills) and model-agnostic biological objectives (Playbooks), seamlessly translating free-form biological requests into reliable execution and rigorous downstream interpretation. Evaluated on a benchmark of 54 query cases derived from published S2F workflows, S2F-Agent systematically outperformed general-purpose LLMs, demonstrating superior reliability accuracy in routing, groundedness, and end-to-end task execution success. We further demonstrate the robustness and scalability of S2F-Agent across model adaptation, variant interpretation, genome-scale functional profiling and personal-genome analysis. First, the agent autonomously adapts a genomic foundation model to quantitative chromatin profiles, resolving sequence features associated with primed and active regulatory states. Second, integrating multi-perspective variant effect predictions prioritized 42 high-priority candidate variants among CAD-associated variants (>16,000), and identified tissue-resolved regulatory mechanisms including the hepatic SORT1 axis. Third, genome-scale profiling of multiple traits GWAS atlas variants (>250,000) revealed pervasive context dependence in molecular consequences and regulatory architecture, highlighting the analytical focus toward fine-grained, tissue-specific regulatory variants. Finally, evidence-gated analysis of personal genomes expanded functional hypothesis generation beyond clinically annotated variants to thousands of prioritized candidates per individual while imposing explicit evidence-dependent boundaries on clinical claims. Collectively, these results establish S2F-Agent as a general framework for converting heterogeneous sequence-to-function capabilities into verifiable, scalable, and evidence-aware genomic analyses. By bridging the chasm between LLMs, specialized S2F ecosystems and rigorous genomic science, this framework democratizes the S2F paradigm for unlocking the full potential of these advanced models in real-world discoveries.

bioinformatics↗

bioRxiv2026

De novo design of transmembrane accessory subunits for fold stabilization and expansion

Transmembrane (TM) proteins play essential roles in biology as transporters, ion channels, chaperones, enzymes, and mediators of signal transduction. However, membrane proteins often suffer from inefficient folding and intrinsic instability. Misfolding in cells can cause numerous loss-of-function pathologies. Likewise, denaturation upon purification in the laboratory is a critical barrier to structure determination and characterization of key biochemical mechanisms. Generalizable strategies to stabilize membrane proteins remain limited. Here, we developed an informatics-based de novo design strategy to create synthetic auxiliary subunits that interact with the TM helices of a model pentameric ion channel, thereby bolstering folding while maintaining channel function. Biochemical and structural characterization reveal the synthetic TM subunits can also be used to create larger multi-spanning designer proteins of custom topology. This proof-of-concept motivates the feasibility of computationally designed accessory TM helices as potential pharmacological chaperone "folding correctors" of membrane proteins in disease and as tools in structural biology.

biochemistry↗

bioRxiv2026

Dynamic Fluoroleucine PET Detects Impaired Cardiac Leucine Uptake Before Hypertensive Left Ventricular Hypertrophy Develops

PurposeLeft ventricular hypertrophy (LVH) is a major complication of chronic hypertension and an independent risk factor for cardiovascular morbidity and mortality. There are currently no clinically validated markers available to identify hypertensive individuals at risk for developing LVH. In hearts of hypertensive rats, we previously described metabolic changes that precede LVH development, including in branched-chain amino acid (BCAA) metabolism. This study investigated whether cardiac leucine uptake, measured with dynamic 5-[18F]fluoroleucine positron emission tomography-computed tomography ([18F]FLE-PET/CT), was impaired and could serve as an in vivo marker for hypertension-induced LVH development. ProceduresWe synthesized [18F]FLE following established radiochemistry protocols and performed dynamic [18F]FLE-PET/CT imaging in 3-month-old spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) control rats (n = 4 per group). Cardiac magnetic resonance (CMR) imaging was conducted on the same animals for structural co-registration. A dual-output reversible two-tissue compartment model with spill-over (SP) and partial volume (PV) corrections was developed to quantify the first-pass rate constant (K1) and total distribution volume (Vt = K1/k2) for [18F]FLE. Protein expression of L-type amino acid transporter 1 (LAT1) and branched-chain keto acid dehydrogenase (BCKDH) phosphorylation status were assessed by immunoblotting of isolated heart tissue. ResultsSHR demonstrated markedly lower first-pass leucine uptake rates (K1) and total distribution volumes (Vt) compared with WKY rats, consistent with reduced cardiac BCAA uptake. Concurrently, LAT1 (SLC7A5) expression was significantly reduced in SHR hearts compatible with decreased leucine uptake. Elevated BCKDH phosphorylation at Ser293 in SHR hearts indicated diminished BCKDH enzymatic activity and impaired BCAA catabolism. ConclusionsDynamic cardiac [18F]FLE-PET imaging successfully detects decreased leucine uptake in hypertensive rat hearts at 3 months of age, before LVH is established at 5 months. Reduced cardiac leucine uptake may thus serve as a surrogate marker for impaired cardiac BCAA metabolism and early in vivo indicator of cardiometabolic dysfunction that precedes LVH. The imaging approach holds translational potential for identifying hypertensive patients at risk for LVH progression.

bioengineering↗

bioRxiv2026

Somatic-to-germline transmission of horizontally acquired extrachromosomal circular DNA in Brassica graft chimeras

Whether somatically acquired traits can contribute to heritable variation has remained an open question in plant biology for over a century. Here, using graft-induced periclinal chimeras between Brassica juncea and Brassica oleracea, we identified extrachromosomal circular DNA (eccDNA) as mobile genetic elements capable of crossing histological boundaries and entering the germline. We demonstrated that grafting could promote the horizontal transfer of eccDNA between somatic cell layers, enabling its stable maintenance in recipient tissues and permitting transmission through sexual reproduction across multiple generations. We found that transmitted eccDNA is non-randomly distributed across the genome, preferentially originating from gene-dense regions, and enriched for features associated with molecular persistence, such as inverted repeats, hairpin-forming sequences, and autonomously replicating sequence (ARS) consensus motifs. These genetic regions conferred replication competence, as evidenced by autonomous propagation in an ARS-less heterologous yeast system. Sexual progeny carrying graft-acquired eccDNA exhibited reproducible and lineage-dependent alterations in leaf morphology and drought tolerance, accompanied by coordinated transcriptional reprogramming. Notably, a subset of inherited eccDNA remained transcriptionally active in progeny, producing transcripts absent from self-grafted controls. Our findings establish eccDNA as heritable extrachromosomal elements that link graft-mediated somatic genetic transfer with stable germline transmission, thereby expanding the molecular scope of heritable variation in plants and providing a conceptual framework for graft-based trait transmission.

plant biology↗

bioRxiv2026

Single-Cell Atlas of Renal Cell Carcinoma Brain Metastasis Uncovers Mechanisms of Immune Dysfunction and Resistance

Brain metastasis (BM) in renal cell carcinoma (RCC) remains poorly understood and often resistant to immune checkpoint inhibitors. We generated a large single-nucleus RNA-seq data of RCC BM, profiling 14 BM samples alongside matched extracranial metastases and primary tumors. Tumor cells in BM displayed neuronal infiltration, neural-like adaptation, and marked remodeling of the microenvironment, including expansion of immunosuppressive myeloid cells and depletion of antigen-presenting dendritic cells. Tumor, immune, and stromal cells exhibited metabolic rewiring characterized by fatty-acid metabolism, oxidative phosphorylation, and MYC-driven programs. CD8 T cells showed terminal exhaustion and impaired proliferative capacity, and tertiary lymphoid structures were absent. Spatial profiling of 12 BM samples (13,128 cells) validated key cellular interactions, while ligand-receptor analysis revealed immunoregulatory circuits between tumor, stromal, and immune cells. These findings define BM-specific adaptations that promote immune evasion and resistance, revealing therapeutic vulnerabilities in RCC BM. SIGNIFICANCESingle-nucleus RNA-sequencing profiling reveals tumor, immune, and metabolic adaptations in renal cell carcinoma (RCC) brain metastases, including neuroglial remodelling and immunosuppressive niche formation. These findings identify immune evasion mechanisms that could contribute to therapeutic resistance, providing new avenues for site-specific therapeutic interventions to improve treatment efficacy and outcomes in patients with RCC BM.

cancer biology↗

bioRxiv2026

Iridescence in pterosaur pycnofibers and the evolution of integumentary coloration

The bodies of pterosaurs, the first flying vertebrates, are covered with integumentary filaments (pycnofibres) thought to be homologous to feathers in dinosaurs, but their coloration remains unknown. Here, we report a layered internal arrangement of melanosomes containing a photonic nanostructure within the monofilaments in a previously undescribed specimen of tapejarid pterosaur Sinopterus dongi from the Early Cretaceous Jehol Biota. Optical simulations showed that this structure reflects green to magenta iridescent coloration, confirming the presence of melanosome-based iridescent coloration previously thought to be unique to birds. This finding deepens our understanding of structure/color gamut relationships in amniotes, while supporting further shared characteristics associated with derived genetic and regulatory shifts in archosaurs.

paleontology↗

bioRxiv2026

The variable wheat stripe rust effector AvrYr7 evades Yr7 recognition through sequence and expression polymorphisms

Introductory paragraphWheat provides about 20% of total dietary calories worldside1. Wheat diseases, including wheat stripe (yellow) rust, cause billions of dollars in losses each year2. Wheat stripe (yellow) rust is caused by the fungal pathogen Puccinia striiformis f. sp. tritici (Pst) which is best controlled by fungicide application and disease resistant wheat cultivars3. To-date, there are over 80 catalogued and >10 cloned yellow rust resistance genes (Yr genes)4. Yet our knowledge of corresponding avirulence (Avr) genes lags far behind5-8. The absence of cloned Avrs reflects Psts complex genome and the lack of robust transformation and genetic systems3. Recent advances in generating high-quality genome assemblies and the development of wheat defense assays have addressed these challenges9-11. Here we clone AvrYr7 which is recognized by Yr712. We further identify six additional alleles of AvrYr7 that escape recognition due to non-synonymous genetic variations, transposable element activity, missense mutation, and expression polymorphism. These findings provide critical insights into virulence evolution in one of the worlds most important wheat pathogens.

plant biology↗

bioRxiv2026

Tripeptidyl peptidase II is essential for maintaining cerebrovascular homeostasis of female mice and represents a novel therapeutic target for vascular dementia

Although cerebrovascular impairment is a known common driver of both Vascular dementia (VaD) and Alzheimers disease (AD), the underlying mechanisms remain poorly defined. Substantial evidence has demonstrated that brain-derived estrogen is essential for cerebrovascular health and neuroprotection. Consequently, therapeutic strategies that can replicate the beneficial effects of estrogen replacement therapy (ERT) in the brain while avoiding its peripheral risks are being actively pursued. Here, we demonstrate that tripeptidyl peptidase II (TPP2) is essential for cerebrovascular homeostasis in adult female mice by orchestrating intracellular Ca{superscript 2} distribution and local estrogen biosynthesis. Specifically, TPP2 deficiency triggers a vicious cycle of Ca{superscript 2} imbalance and estrogen deficiency, thereby disrupting the anticipatory unfolded protein response (UPR). This disruption consequently drives aberrant pexophagic flux and ultimately depletes ether-linked phosphatidylcholine (PC-O)--the essential building block of endothelial cell (EC) membranes. In addition, we identified PC-O as a key facilitator of choline uptake through FLVCR2 and therefore its deficiency causes subsequent choline depletion as well as significantly decreased acetylcholine (ACh). Consistent with the fact that choline is a building block of PC and that ACh is a primary driver of vascular dilation, TPP2 depletion leads to significant cerebrovascular degeneration characterized by narrowed lumens, decreased EC number, and abnormal aggregation of ECs within the blood vessel. Additionally, AAV-mediated specific expression of Far1 in ECs to promote ether-PC biosynthesis significantly increases cerebrovascular volume and diameter in hippocampi of adult female TPP2 knockout (T2KO) mice. Importantly, ectopic expression of Far1 in ECs not only significantly ameliorates memory impairment in global and conditional TPP2-depleted female mice, but also significantly improves memory performance of naturally aged female mice. In total, our findings establish TPP2 as a key determinant of cerebrovascular homeostasis in adult female mice, positioning it as a novel therapeutic candidate for the treatment of VaD and AD.

neuroscience↗

bioRxiv2026

Parallel Activation and Interference CRISPR (PAIR) with Sequencing Uncovers DNA Repair Networks Guiding Precision Cell Engineering

The dynamic balance of cellular homeostasis is often maintained by opposing regulatory pathways, yet most genetic screens interrogate them in one direction and therefore miss the bidirectional gene-gene interactions that shape complex phenotypes such as DNA damage response (DDR). Here, we present PAIR (Parallel Activation and Interference CRISPR), a bidirectional perturbation platform that enables simultaneous activation and suppression of distinct genes within the same cell using CRISPR activation (CRISPRa) and Cas13d RNA knockdown. Applying PAIR to the CRISPR/Cas9 induced DSB repair screen, we mapped gene-gene interactions across competing repair branches and identify synergistic perturbations, including NBN activation combined with suppression of end-joining factors, that shift repair outcomes toward homology-directed repair (HDR) and improve the precision of CRISPR-based gene editing. Using coupled PAIR with single-cell transcriptomic, we further demonstrated that NBN activation induces inflammatory and interferon programs, whereas co-suppression of end-joining factors buffers this response, revealing transcriptional states missed by conventional unidirectional perturbations. To translate these findings into non-viral chimeric antigen receptor (CAR) T cell engineering, we developed an mRNA-based strategy for parallel overexpression and knockdown of NBN-anchored DDR effectors in primary T cells, priming the T cells into a transient HDR-favored state that enhances the efficiency of CAR knock-in on the TRAC locus. Together, the PAIR system provides a general framework for studying opposing regulatory networks, uncovering hidden cell states, and guiding cell-state engineering through bidirectional perturbation.

bioengineering↗

bioRxiv2026

IFITM1 inhibits Henipavirus membrane fusion by trapping ephrinB2 receptors in fusion-unfavorable membrane nanodomains.

Nipah (NiV) and Hendra (HeV) viruses are prototype henipaviruses (HNV) within the family of Paramyxoviridae. They are highly pathogenic zoonotic paramyxoviruses that cause fatal encephalitis in humans, with case fatality rates of 40-70%. These viruses use the receptor-binding protein (G) to engage host receptors ephrinB2 and/or ephrinB3, while the fusion protein (F) drives membrane fusion leading to virus entry and cell-cell fusion. Interferon-inducible transmembrane (IFITM) proteins restrict the infection of many unrelated viruses at virus entry. We show that the knockdown of IFITM1 enhances HNV entry and cell-cell fusion in both human epithelial and endothelial cells, while overexpression of IFITM1 inhibits both. Using single-molecule localization microscopy, we observed that IFITM1 forms nanoclusters on the plasma membrane that are enhanced upon interferon stimulation. IFITM1 clusters spatially co-occur with GM1-enriched, raft-like membrane domains. Notably, IFITM1 interacts with ephrinB2, and redistributes ephrinB2 into larger and denser clusters, potentially within GM1-enriched regions. Using single-particle tracking, we further show that ephrinB2s lateral diffusion on the plasma membrane is confined by IFITM1, and this confinement is not reversed by amphotericin B, a membrane-permeabilizing agent that increases membrane fluidity. Our data suggest that IFITM1 sequesters ephrinB2 in fusion-unfavorable membrane domains, thereby enhancing the energy barrier for virus-cell membrane fusion and decreasing the virus-receptor encounters. SignificanceNipah (NiV) and Hendra (HeV) viruses are zoonotic paramyxoviruses that cause fatal encephalitis in humans. Here, we show that IFITM1 inhibits NiV and HeV entry at the surface of epithelial and endothelial cells. We report that IFITM1 restricts NiV entry by sequestering the host receptor ephrinB2 in membrane domains that are energetically unfavorable for virus-cell membrane fusion. IFITM1 also restricts ephrinB2 lateral diffusion on the plasma membrane, thereby reducing productive virus-receptor encounters. Notably, amphotericin B treatment, which increases membrane fluidity, did not rescue IFITM1-mediated confinement of ephrinB2 or restore NiV entry. Our data reveal a new mechanism by which IFITM1 restricts NiV infection, suggesting that its antiviral activity extends beyond simply altering membrane fluidity.

microbiology↗

bioRxiv2026

Serine Stabilizes SLC7A11 and Enhances Cystine Influx to Protect Against Acute Pancreatitis

Lethal sterile inflammatory diseases are linked to amino acid metabolism, but the role of serine remains unclear. Here, we show that dysregulated serine metabolism and reduced plasma serine levels correlate with disease severity of acute pancreatitis (AP) in patients and mouse models. Elevating serine levels via exogenous serine supplementation or pancreatic phosphoglycerate dehydrogenase (PHGDH) overexpression mitigates pancreatic injury, whereas a serine deprivation diet or pancreatic PHGDH knockdown exacerbates AP. Serine prevents cell death and oxidative stress in pancreatic acinar cells, human induced pluripotent stem cells-derived pancreatic organoids and mouse pancreatic tissue. Serine enhances cysteine and glutathione biosynthesis primarily by promoting solute carrier family 7 member 11 (SLC7A11)-dependent cystine uptake rather than by serving as a direct substrate. Mechanistically, the E3 ubiquitin ligase NEDD4 mediates ubiquitination and degradation of SLC7A11, whereas serine binds to NEDD4 and thereby inhibits SLC7A11 degradation. Similarly to serine, pharmacological inhibition of NEDD4 alleviates lipid peroxidation and pancreatic injury. These findings identify serine as a critical signaling regulator of SLC7A11 stability and oxidative stress, and provides a new therapeutic strategy for AP and associated sterile inflammatory disorders. HighlightsAcute pancreatitis (AP) is linked to abnormal serine metabolism and serine depletion. Serine prevents cell death in AP acinar cells, human pancreatic organoids and mice. Serine promotes SLC7A11-dependent cystine uptake and glutathione levels in acinar cells. Serine reduces NEDD4-mediated ubiquitination of SLC7A11. In briefSerine protects against cell death and pancreatic injury in acute pancreatitis by stabilizing SLC7A11 through disruption of NEDD4-mediated ubiquitination in acinar cells.

pharmacology and toxicology↗

bioRxiv2026

Hippocampal Glucuronyl C5-epimerase promotes stress resilience by directly engaging PI3K through a non-enzymatic mechanism

Major depressive disorder (MDD) seriously affects human physical and mental health and causes global socioeconomic burdens. Stress-induced depressive etiology is linked to dendritic atrophy in the hippocampus, however, the underlying mechanism was poorly understood. Here, we identify that glucuronyl C5-epimerase (Glce), a heparan sulfae-modifying enzyme, as a critical regulator of hippocampal dendritic integrity and stress resilience. Genetic ablation of Glce in the hippocampal excitatory neurons is sufficient to induce dendritic atrophy and depressive-like behaviors, whereas its restoration rescues these deficits. Notably, Glce levels are reduced in the plasma of depressed individuals. We further find that Glce functions independently of its enzymatic activity, instead binding directly to and activating PI3K (p85/p110), thereby triggering an Akt/CREB/BDNF signaling cascade. Together, our findings uncover a non-canonical role for Glce and establish that Glce-PI3K-BDNF axis is essential for maintaining hippocampal structure and behavioral resilience, thereby highlighting a critical role of Glce in the pathophysiology of depression.

neuroscience↗

bioRxiv2026

3D-PAULM: Integrated Photoacoustic Tomography and Ultrasound Localization Microscopy for Multiparametric Brain and Tumor Imaging

Understanding processes such as blood-brain barrier (BBB) disruption and tumor progression can greatly benefit from simultaneous molecular, functional, and hemodynamic imaging in deep tissue, yet few existing imaging modalities can provide all three in a single system. Here, we present an integrated imaging platform that combines 3D photoacoustic tomography with ultrasound localization microscopy (3D-PAULM) to enable intrinsically co-registered, multiparametric imaging. 3D-PAULM unifies multispectral photoacoustic molecular imaging, ultrasound B-mode imaging, microbubble-enhanced power Doppler, and ultrasound localization microscopy, and concurrently measures blood oxygenation, blood perfusion, microvascular flow dynamics, and molecular probes from near-infrared dyes and photoswitchable phytochromes. We apply 3D-PAULM to quantify BBB leakage in focal ischemia and systemic inflammation, and to perform high-sensitivity molecular imaging of solid tumors alongside functional mapping of tumor hypoxia and super-resolved vascular remodeling. Together, these results establish 3D-PAULM as a versatile platform for integrated functional and molecular imaging in deep tissue.

bioengineering↗

bioRxiv2026

Targeting the AgMOBP1-PfSyn5 mosquito-parasite interface completely blocks malaria transmission

Malaria control remains constrained by an incomplete understanding of the molecular interactions that enable Plasmodium falciparum transmission within mosquito vectors. Although blood-stage parasite biology has been extensively studied, the mosquito-stage processes required for transmission are less well defined. Here, we characterize a previously unannotated Anopheles gambiae midgut factor, Midgut Ookinete Binding Protein 1 (AgMOBP1; AGAP008138), and define its role in parasite infection. AgMOBP1 is a membrane-associated protein localized predominantly to the apical surface of midgut epithelial cells, is induced by blood feeding, and appears restricted to anopheline mosquitoes. Functional infection assays showed that exogenous recombinant AgMOBP1 enhanced Plasmodium infectivity by increasing midgut oocyst burden, whereas anti-AgMOBP1 antibodies significantly reduced infection, establishing AgMOBP1 as a host factor that promotes transmission. Using biochemical interaction assays, we identified the parasite binding partner of AgMOBP1 as the P. falciparum syntaxin-5-like protein PfSyn5. Disruption of this interaction with anti-PfSyn5 polyclonal antibodies produced exceptionally potent transmission-blocking activity, with complete inhibition at 1 {micro}g/mL and an estimated IC50 of [~]3 ng/mL ([~]20 pM). This potency is approximately one million-fold lower than the total IgG concentration in normal human plasma. Together, these findings identify a critical mosquito-parasite molecular interface required for malaria transmission and establish the AgMOBP1-PfSyn5 axis as a promising target for transmission-blocking vaccines, monoclonal antibodies, and other intervention strategies. Significance StatementInterrupting malaria transmission requires a deeper mechanistic understanding of how Plasmodium parasites interact with their mosquito vectors. Here, we define a previously uncharacterized mosquito midgut factor, AgMOBP1, and identify its parasite binding partner, the Plasmodium falciparum membrane protein PfSyn5. We show that this molecular interaction is required for efficient parasite transmission to Anopheles mosquitoes. Remarkably, antibodies against PfSyn5 completely blocked transmission at low concentrations, revealing exceptional functional potency. These findings uncover a previously unknown mosquito-parasite invasion pathway and establish the AgMOBP1-PfSyn5 interface as a high-value target for transmission-blocking vaccines, monoclonal antibodies, and next-generation malaria control strategies.

microbiology↗

bioRxiv2026

Transformed reactivation of latent working memory enables hierarchical language processing

Human language comprehension requires tracking words across intervening material to construct grammatical structures, yet two fundamental questions remain unresolved: whether maintenance of earlier words relies on sustained neural activity or activity-silent working memory mechanisms, and whether memory retrieval during integration engages domain-general or language-selective networks. Using magnetoencephalography with time-resolved decoding, we tracked main-clause subjects as participants heard sentences with embedded clauses (e.g., "The dog, who chases the cat, jumps over the mud."). The subject representation ("dog") decayed to baseline during the embedded clause but reactivated after the main-clause verb ("jumps"), with transformed rather than reinstated neural codes. Critically, reactivation emerged first in right dorsolateral prefrontal cortex before engaging frontotemporal language regions, and reactivation strength was modulated by syntactic structure and predicted comprehension accuracy. These findings demonstrate activity-silent working memory maintenance, structure-dependent retrieval, and cooperative function of domain-general and language-selective networks during hierarchical language processing.

neuroscience↗