Extracellular vesicles (EV) serve as critical mediators in physiological and pathological processes, holding great promise for cancer diagnosis, real-time monitoring, and prognostic applications. However, their small size and low density present considerable challenges in achieving efficient, specific, and mild isolation, which currently hinders widespread clinical translation. Here, we report a proximity-induced pH-responsive DNA switch (PPS) that enables reversible EV capture via Hoogsteen triplex formation. Specifically, dual-aptamer probes targeting EpCAM and CD63 on EV exploit membrane fluidity to form proximity-induced duplex helix, which assemble into Hoogsteen triplex helix with magnetic bead-conjugated strands at mildly acidic conditions (pH6.5) for EV capture. Crucially, simply adjusting the pH to neutral (pH7.4) triggers triplex dissociation, releasing intact EV without using chemical denaturants and preserving vesicle integrity for high-purity isolation. Under optimized conditions, this method achieves 79.4% and 72.6% EV purification efficiency in PBS and complex biological matrices, respectively, within 1 h. Furthermore, its modular aptamer design enables rapid adaptation to diverse EV subpopulations through simple probe substitution, without modifying the core framework, thereby reducing cost and complexity for broad applications. Meanwhile, due to the gentle capture-release process, the structural integrity and bioactivity of EV are well preserved, as demonstrated by wound-healing and cellular uptake assays. These advantageous features─rapid processing, high specificity, mild operation, and preservation of EV activity─indicate that the PPS strategy is a robust, nondestructive method for EV isolation. It thus holds significant potential for further application in diverse EV-related research fields such as disease diagnosis and drug delivery.
The anaerobic co-digestion (AcoD) of food waste (FW) and biodegradable plastics (BPs) is a promising waste-to-energy strategy, yet it remains severely bottlenecked by asynchronous hydrolysis. Overcoming this limitation via traditional trial-and-error optimization is prohibitively slow and expensive. To address this, an integrated machine learning (ML) framework was developed, coupling predictive modeling with rigorous experimental and mechanistic validation. Virtual screening initially identified mesophilic enzyme-loaded biochar (BC) as the optimal intervention. Consequently, a novel proteinase K-loaded BC (PKBC) was synthesized, which successfully achieved up to 90% degradation of 2-mm BPs films in a 120-day trial. A high-precision random forest model further mapped the system dynamics, confirming that degradation is positively driven by reaction time and methane yield, but tightly constrained by larger particle sizes. Providing a fundamental basis for these predictions, multi-omics analyses revealed that PKBC triggers a targeted metabolic cascade. This cascade reprograms the microbial community to accelerate lactate-driven BPs cleavage and maximize hydrogenotrophic methanogenesis. Ultimately, this work delivers a highly efficient solution for mixed waste treatment and establishes a transferable AI-first paradigm for the intelligent design of complex bioprocesses.
Polymer gels are fundamentally constrained by an inherent trade-off between mechanical stiffness and dynamicity, impeding the synergistic integration of high strength with adaptive functionalities such as self-healing, recyclability, and reconfigurability. Here, we present a strategy to resolve this long-standing challenge by engineering switchable hydrogen-bond clustering (HC) within a deep eutectic solvent-based adhesive gel. Precise modulation of water content triggers the reversible transition between a rigid state (elastic modulus ∼45 MPa) and a highly dynamic state (modulus reduction by 105-fold). This on-demand switching capability facilitates direct 3D printing, efficient recycling assisted by trace water, and autonomous self-healing (∼83% recovery efficiency). Crucially, the gel retains both high stiffness and robust interfacial adhesion after turning on the HC. The HC switchable strategy establishes a versatile design principle for fabricating strong, yet dynamically reconfigurable polymeric networks, with promising implications for advanced wearables, soft robotics, and adaptive adhesive technologies.
Monitoring therapeutic response is essential for improving the precision and efficacy of photodynamic therapy (PDT), yet quantitative and real-time evaluation remain challenging. Herein, we report a programmable DNA nanotube-based dual-mode biosensing platform for monitoring PDT-induced apoptosis through complementary fluorescence and electrochemical signals. In this system, methylene blue (MB) functions as a multifunctional component, simultaneously acting as a photosensitizer, near-infrared imaging probe, and electrochemical reporter. A caspase-3-responsive peptide is integrated into the DNA nanotube scaffold, enabling apoptosis-triggered fluorescence activation. Caspase-3 activation triggers peptide cleavage, resulting in fluorescence recovery and enhanced electrochemical signals, with limits of detection (LOD) of 0.786 ng/mL for the fluorescence method and 0.622 ng/mL for square-wave voltammetry (SWV). Gold nanoparticles incorporated into the DNA nanotube further catalyze endogenous H2O2 decomposition to generate oxygen, alleviating tumor hypoxia and enhancing PDT efficacy. The platform demonstrated accurate dual-mode detection capability in complex biological samples. These results demonstrate that the programmable DNA nanotube-based platform enables synchronized electrochemical and fluorescence biosensing through a shared caspase-3-responsive mechanism, thereby allowing real-time therapeutic feedback during photodynamic therapy.
The antiviral protein MORC3 is frequently inhibited by viruses. To counteract viral antagonism, MORC3 represses a noncanonical pathway of type-I-interferon (IFN) such that viral inhibition of MORC3 triggers ( > 10,000-fold) IFN induction. How MORC3 represses this pathway, and why IFN induction upon MORC3 loss is so potent without canonical IRF3/7 transcription factors, is unknown. Here, we show that MORC3 restricts chromatin accessibility at tandem repeat elements harboring up to 61 homotypic transcription factor motifs. One such element becomes a potent enhancer of IFNB1 upon MORC3 loss. Its motif cluster contains 45 PU.1 binding sites and is necessary and sufficient for MORC3-mediated repression and enhancer activity upon MORC3 loss. PU.1 recruits MORC3 to repress this enhancer by recruiting DAXX and enabling H3.3 incorporation. Upon MORC3 loss, PU.1 drives IRF3/7-independent IFN induction. Other restricted tandem repeats contain homotypic motif clusters of SPI, AP-1, and SP/KLF transcription factors. Our findings uncover a TF motif cluster-driven repression mechanism by MORC3 at tandem repeats, enabling specific repression of an IFNB1 enhancer such that viral antagonism of MORC3 induces interferon.
Endometriosis defined by the growth of endometrial tissues outside the uterus, affects women of reproductive age. A critical process in endometriosis progression, angiogenesis involves endothelial cell migration, proliferation and tube formation, with vascular endothelial growth factor (VEGF) playing a powerful role. Exposure to endocrine-disrupting pollutants, like Hexachlorobenzene (HCB) andChlorpyrifos (CPF), is linked to a higher risk of reproductive diseases including endometriosis. Both HCB and CPF are weak aryl hydrocarbon receptor (AhR) ligands. These study examined HCB and CPF mechanisms of action in endometriosis-associated angiogenesis in vitro. Our results show that HCB (0.005, 0.05 and 5 μM) and CPF (0.5-50 μM) induced VEGF secretion in human stromal endometrial cells (T-HESCs). Moreover, HCB- or CPF-conditioned media (HCB-CM, CPF-CM) from T-HESCs boosted endothelial cell (EA.hy926) proliferation and survival. In addition, wound healing assays rendered an increase in EA.hy926 cell migration after exposure to HCB-CM (0.005-0.5 μM) or CPF-CM (5 μM), while tube-like structures revealed an increase in neovasculogenesis after HCB-CM (0.5 μM) or CPF-CM (5 μM). Our findings show that HCB- and CPF-induced angiogenesis was mediated by AhR and VEGF receptor-2. These results demonstrate that both pesticides increase VEGF secretion in endometrial cells and triggers angiogenesis, a critical event in endometriosis progression.
Recently, ferroptosis has emerged as a pathogenic mechanism that drives metabolic dysfunction-associated steatohepatitis (MASH); however, the upstream triggers and their relevance to fibrosis remain poorly understood. Here we identified dietary cholesterol-induced ferroptosis and the downregulation of peroxisome proliferator-activated receptor delta (PPARδ) as central drivers of MASH pathogenesis. To investigate this, human liver samples and cholesterol-enriched dietary murine models of MASH were examined in parallel with mechanistic studies in hepatocytes and hepatic stellate cells (HSCs). Cholesterol-induced MASH was associated with pronounced hepatic lipid peroxidation and the selective downregulation of PPARδ. The loss of PPARδ disrupted redox homeostasis and sensitized hepatocytes to ferroptosis, whereas exosomal double-stranded DNA released from ferroptotic hepatocytes activated STING-TBK1-IRF3 and induced expression of profibrotic genes in HSCs. These effects were reversed by either overexpression of hepatocyte-specific PPARδ or pharmacologic treatment with DN203316, a novel and highly selective PPARδ agonist. In vivo, DN203316 mitigated ferroptosis, inflammation and fibrosis without inducing metabolic derangement. These findings were substantiated by clinical data demonstrating a marked increase in lipid peroxidation and STING-driven HSC activation in liver tissues from patients with MASH. In conclusion, PPARδ is a key regulator of cholesterol-induced ferroptosis and exosome-mediated fibrogenic signaling in MASH. DN203316 offers a promising therapeutic strategy to suppress ferroptosis, disrupt hepatocyte-HSC crosstalk and attenuate disease progression.
Obesity stems from a chronic imbalance between energy intake and expenditure. Current therapeutic strategies primarily focus on reducing caloric intake, yet their long-term efficacy is often limited by compensatory metabolic adaptations that lead to weight regain. This review outlines the neural mechanisms through which the central nervous system regulates appetite and the peripheral metabolic pathways that drive adipose thermogenesis. Furthermore, it examines how integrated approaches-spanning from approved to preclinical and clinical-stage investigational agents (e.g., dual- or multi-target agonists), microbiome-targeted interventions (e.g., probiotics), and exercise therapy-can synergistically overcome the limitations of single-pathway strategies. Ultimately, this review provides a theoretical foundation for designing next-generation, personalized, multimodal obesity management regimens. Traditional weight-loss drugs primarily act by centrally suppressing appetite, reducing food intake through modulation of neural circuits in regions such as the hypothalamus. However, studies show that relying on appetite suppression often triggers compensatory metabolic adaptation, ultimately leading to weight regain. Current anti-obesity drug development is therefore shifting toward integrated central-peripheral dual mechanisms. GLP‑1/glucagon dual-receptor agonists and triple-receptor agonists (such as retatrutide) have exhibited unprecedented weight-loss efficacy in clinical trials. These novel agents overcome the limitations of single-target appetite suppression by synergistically integrating central anorexigenic signaling with peripherally mediated increases in energy expenditure, thereby achieving more potent and durable weight reduction. The sustainability of obesity treatment relies on a dual-pronged intervention strategy: suppressing appetite to reduce energy intake while actively promoting energy expenditure, thereby overcoming the metabolic adaptation and weight rebound associated with monotherapy.
Cellular decision-making relies on the integration of multiple extracellular cues into coordinated functional responses. Synthetic biology provides tools to rewire this process by engineering receptors that convert defined inputs into programmable outputs. Here, we describe a synthetic receptor-based architecture that enables monocytic-like cells to sense an immune-regulatory ligand and conditionally activate a phagocytic program. We engineered a synthetic Notch-based receptor (SNIPR) that detects programmed death-ligand 1 (PD-L1), a broadly expressed immune-regulatory ligand. Upon PD-L1 engagement, the circuit triggers programmable outputs, including expression of a fluorescent reporter or CV1-Fc, as model effector that interferes with CD47-mediated inhibition of phagocytosis. We show that circuit activation scales with PD-L1 levels, partially attenuates PD-1/PD-L1 signaling, and that conditional CV1-Fc expression enhances engulfment of SKOV-3 ovarian cancer cells by THP-1-derived macrophages in vitro. Collectively, this work reframes PD-L1 from an end-point therapeutic target to a programmable input signal for synthetic circuit activation and establishes a modular framework for ligand-responsive control of engineered macrophage behaviour.
Programmable technologies that sense nucleic acid signatures in living cells and trigger cellular functions hold promise for biotechnology and medicine. Here, we develop SONAR (Sensing Of Nucleic acids using ASOs and Reverse-transcriptases), a platform that detects target DNA and RNA sequences and triggers controlled gene expression in human cells. SONAR operates through circularizable single-stranded DNA (ssDNA) sensors that, upon hybridization with complementary DNA or reverse-transcribed RNA, undergo target-dependent ligation via cellular ligases, subsequently driving expression of genetic payloads. For RNA sensing, we employ antisense oligonucleotides (ASOs) to prime targeted reverse transcription, generating complementary DNA that promotes ssDNA circularization. We demonstrate SONAR's ability to detect ssDNA, exogenous and endogenous RNA, couple sensing to programmable expression of diverse protein payloads, including reporters, recombinases, and genome editors, and enable enrichment and clonal recovery of target-positive cells from mixed populations. This platform establishes a versatile framework for targeted nucleic acid detection and inducible gene expression, with broad potential applications in diagnostics, therapeutics, and synthetic biology.
Anaphylaxis during pregnancy is a rare but potentially life-threatening condition for both mother and fetus, requiring rapid recognition and immediate treatment. Although the fundamental mechanisms of anaphylaxis in pregnancy are similar to those in nonpregnant women, physiological adaptations of pregnancy, peripartum exposures, and fetal considerations substantially complicate diagnosis, management, and prevention, contributing to variability in care and avoidable adverse outcomes. In this multidisciplinary review, experts in allergy-immunology, obstetrics, anesthesiology, and epidemiology synthesize current evidence on the epidemiology, triggers, pathophysiology, diagnostic challenges, management, outcomes, and prevention of anaphylaxis throughout pregnancy, labor, and delivery. We highlight how gestational cardiovascular and respiratory changes may obscure classic diagnostic features, emphasize the safety and critical importance of prompt intramuscular epinephrine use as first-line therapy, and review maternal and fetal outcomes associated with timely versus delayed intervention. Strategies for risk stratification, allergology workup, prevention of recurrence, and implementation of coordinated care pathways are discussed. This review underscores the need for increased awareness, structured interdisciplinary collaboration, and integration of prevention-focused strategies across obstetric and allergy care. By providing a practical, evidence-based framework, it aims to support health professionals in optimizing diagnosis, management, and maternal-fetal safety when anaphylaxis occurs during pregnancy.
Perinatal brain injury (PBI) is a major predictor of neurological disability. Commonly associated with prematurity, infection, stroke, hypoxia-ischemia, hemorrhage, and/or toxin exposure, PBI triggers acute and persistent systemic inflammation. There are many stages of vulnerability to PBI during development including pregnancy, birth - term and preterm, and neonatal age. The vulnerable stages can compound inflammation through injury to the placental-fetal-brain axis, adaptive and innate immune system development, neural-immune communication, and central nervous system maturation. Neonates exhibit unique inflammatory signatures and lasting neural-immune responses to various etiologies. Chronic immune dysregulation and priming to a secondary, later-in-life immune challenge defines different forms of PBI while shaping the neonatal and adult immune response with long-term changes. Immunomodulated changes impact regulatory, helper and innate T cells, neutrophils, natural killer cells and immune responsiveness. The major routes of persistent and compounding inflammation in PBI are perinatal neural-immune interactions, cytokine influx, and glial crosstalk. Most treatments are not administered long enough or in the optimal time window to combat sustained inflammation in tertiary and quaternary phases of PBI pathophysiology and are ineffective in reducing neonatal mortality and morbidity and promoting functional recovery. Indeed, persistent systemic and central inflammation is a likely explanation for failed recovery of PBI after the resolution of acute insults. We propose attenuating persistent inflammation and normalizing systemic immune reactivity as key to reducing the functional impact of PBI throughout the lifespan through various avenues including therapeutic treatment, gut microbiome modulation, and novel immunomodulation from preclinical research.
Osmotic stress-induced Ca2⁺ accumulation promotes the formation of specific RFP-ATG8i labeled autophagosome near the hypocotyl-root transition zone, suggesting ATG8i-dependent autophagy or reticulophagy, as a mechanism to alleviate osmotic and endoplasmic reticulum stress in plants. Autophagy is a crucial and evolutionarily conserved process that breaks down and recycles damaged or unnecessary cytoplasmic components. This process is essential for plant growth and for responding to environmental stresses. In this study, we investigated how autophagy is regulated during osmotic stress in Arabidopsis thaliana, with a focus on the behavior of the RFP-tagged ATG8i protein and the potential role of cytosolic Ca2⁺ in this process. Osmotic stress was induced with sorbitol and Ca2+ accumulation was monitored with a Cameleon transgenic line. Ca2⁺ influx from the apoplast was disrupted using EGTA, and autophagy was analyzed in a transgenic line expressing RFP-ATG8i under its native promoter. We found that osmotic stress triggers Ca2⁺ accumulation, with a pronounced response in the hypocotyl-root transition zone and a weaker response at the root tip. Both osmotic stress and Ca2⁺ signaling promote the accumulation of RFP-ATG8i-labeled autophagosomes in predominantly in hypocotyl-root transition zone. Furthermore, analysis of GFP-HDEL/RFP-ATG8i doubled transgenic line revealed colocalization of RFP-ATG8i with the endoplasmic reticulum marker HDEL, suggesting that ATG8i participates in reticulophagy and may contributed to the ER turnover under stress conditions.
Diabetic retinopathy (DR) and retinal aging, though arising from distinct causes, share converging mechanisms-including oxidative stress, chronic inflammation, mitochondrial dysfunction, and AGE accumulation-that compromise retinal integrity. These overlaps suggest common gene expression patterns, and highlight the contribution of disease-associated or accelerated aging processes to diabetes-induced retinal injury. We systematically retrieved DR- and aging-related human genes from public genetic databases, identified their overlap, and focused on those involved in key metabolic pathways (AGEs, oxidative stress, lipid metabolism, and autophagy). Genes were then cross-validated across multiple databases and filtered by ocular expression to ensure relevance to retinal pathology. Our findings show that although DR and retinal aging arise from distinct etiologies, they converge on four principal metabolic pathways-oxidative stress, AGE accumulation, lipid dysregulation, and impaired autophagy-that collectively drive similar vascular, neuronal, and inflammatory injury within the retina. Shared genes with high ocular expression reinforce the biological relevance of these pathways, while multi-pathway hub genes appear to function as central regulators that integrate redox imbalance, metabolic disruption, and proteostatic failure. These results provide a unified molecular perspective of retinal degeneration and support the potential development of therapeutic approaches designed to simultaneously target age- and diabetes-associated retinal pathology. This review suggests that DR and retinal aging, although initiated by distinct triggers, converge on shared metabolic pathways-oxidative stress, AGE accumulation, lipid dysregulation, and autophagy impairment-mediated by genes expressed in ocular tissues. Within these intersecting pathways, shared hub genes emerge as central control nodes that may amplify molecular dysfunction and represent potential therapeutic entry points. By mapping these molecular intersections, this study may provide a unified mechanistic perspective of retinal degeneration and support the development of dual-purpose strategies aimed at preventing or mitigating both DR and aging-related retinal decline. These findings highlight potential translational opportunities for targeting shared metabolic networks; however, they should be interpreted as hypothesis-generating and require further experimental and clinical validation to establish causal relationships and therapeutic relevance.
Cancer and cardiovascular diseases represent leading chronic threats to human health, with pharmacotherapy serving as the primary intervention for both. Plant-derived bioactive compounds have emerged as vital sources of antineoplastic agents. Veratramine (VEM), a naturally occurring plant steroidal alkaloid, is widely used in medicine and agriculture. Assessing the toxicity of natural compounds to living organisms is crucial, given the pivotal role of the cardiovascular system in maintaining physiological homeostasis. Nevertheless, the potential toxicity of natural compounds to this vital system remains poorly understood. Herein, we used zebrafish as a model system to evaluate the effects of VEM on cardiac development. Our results demonstrated that VEM impairs zebrafish larval development, with cardiovascular toxicity manifested as yolk sac edema, pericardial edema, increased heart size, reduced atrial-ventricular overlap, increased distance between the sinus venosus and bulbus arteriosus, abnormal cardiac looping, cardiac ejection disorders, and decreased heart rate. Specifically, VEM dysregulates the expression of genes involved in cardiac development, induces oxidative stress in the cardiac region of zebrafish larvae, and triggers cardiomyocyte apoptosis and abnormal proliferation, ultimately leading to cardiac injury. Furthermore, our study is the first to show that VEM exposure activates the Wnt/β-catenin pathway and treatment with a Wnt signaling inhibitor rescues both VEM-induced cardiac developmental defects and oxidative stress. Taken together, our findings indicate that VEM induces cardiac developmental defects in zebrafish larvae by inducing oxidative stress and activating the Wnt/β-catenin pathway.
Salmonella is an intracellular pathogen that can reside within a vacuole, which protects it from cytosolic host defenses at the expense of limited nutrient access. Glucose serves as a critical carbon source supporting Salmonella's intracellular replication. However, the molecular mechanisms driving glucose uptake of host cells and the pathways by which cytosolic glucose becomes accessible to intravacuolar Salmonella remain poorly understood. Here, we elucidate a three-pronged strategy through which Salmonella Typhimurium (S. Typhimurium) exploits Glut1 to co-opt host glucose metabolism for pathogenic advantage. Firstly, S. Typhimurium infection upregulates the glucose transporter Glut1 by activating the MAPK signaling cascade, enhancing host glucose uptake, and accelerating glycolytic flux. Secondly, S. Typhimurium utilizes Glut1 to the bacterial vacuolar membrane to establish a glucose-import conduit that facilitates bacterial acquisition of cytosolic glucose. Thirdly, K29-linked ubiquitination on bacterial vacuolar membranes is a previously unrecognized regulatory mechanism that potentiates Glut1 transporter activity. Inhibition of Glut1 potentiates S. Typhimurium-triggered innate immune responses and attenuates bacterial virulence in vitro and in vivo. Collectively, these findings delineate a novel paradigm of metabolic hijacking, wherein S. Typhimurium systematically rewires host glucose metabolic networks to support intracellular proliferation, providing new insights into host-directed antimicrobial interventions.
The hepatitis E virus (HEV) capsid protein, pORF2, mediates virion assembly, attachment and entry, yet the molecular mechanisms underlying these processes remain poorly defined. Structural studies have provided high-resolution views of pORF2 virus-like particles, revealing an architecture like that of the calicivirus VP1. However, while a paradigm has been established for calicivirus capsid dynamicity - environmentally triggered transitions that regulate receptor engagement, uncoating and immune evasion - comparable conformational plasticity has not yet been explored for HEV. At the same time, efforts to map pORF2 interactions with host factors have yielded several candidate attachment and entry molecules, but no definitive receptor. This review summarises the current knowledge of the structure, forms and host interactions of pORF2, contrasting it with the extensive body of work that has revealed the dynamic behaviour of the calicivirus capsid. This 'dynamic capsid lens' view of HEV may inspire new approaches to unresolved aspects of HEV entry biology. These include how pORF2 engages with host factors, how quasi-enveloped and non-enveloped particles differ functionally, and whether environmental cues encountered during gut-to-liver transit affect capsid conformation. To determine whether HEV, like its calicivirus relatives, exploits capsid dynamicity to establish infection, it will be key to integrate structural, biophysical and cell-based approaches.
2,2',3,4,4',5'-Hexachlorobiphenyl (PCB138) is a persistent organic pollutant with potential intestinal toxicity. Theabrownin (TB) can improve intestinal injury, but its protective effect against PCB138 remains unclear. This study found that 48-week PCB138 exposure induced ileal damage, inflammation, oxidative stress, and necroptosis in mice, which were mitigated by TB intervention. TB alleviated PCB138-induced intestinal damage by suppressing oxidative-stress-mediated intestinal epithelial cell necroptosis. Mechanistically, both TB and N-acetylcysteine (NAC) suppressed PCB138-triggered ROS and subsequent RIPK1/RIPK3/MLKL phosphorylation. The additive effect of NAC combined with necrostatin-1 confirmed ROS as the upstream trigger and RIPK1 as the downstream executor. TB acts similar to NAC, establishing that it prevents necroptosis by scavenging upstream ROS. This study revealed a new mechanism of PCB138-induced intestinal toxicity and identified TB as a potential protective agent against environmentally pollutant-triggered intestinal inflammation.
Anaerobic chain elongation (CE) has emerged as a promising technology for upgrading low-value organic substrates into high-value medium-chain fatty acids (MCFAs); however, achieving targeted metabolic flux and efficient electron transfer remains challenging. To address this, this study explores the role of iron speciation in enhancing chain elongation (CE) driven by ethanol. Two iron-modified activated carbons, Fe3O4@AC and ZVI@AC, were evaluated to assess their impact on microbial metabolic networks. Results revealed that Fe3O4@AC significantly enhanced caproate production (4600.0 mg/L) and electron transfer efficiency (87.0 %), while ZVI@AC triggered a diversion towards alcohol production (940.61 mg/L n-butanol). The superior performance of Fe3O4@AC was attributed to its semiconductive properties, which facilitated interspecies electron transfer (potentially via DIET-like mechanisms) and balanced electron flow, promoting the activation of both fatty acid biosynthesis (FAB) and reverse β-oxidation (RBO) pathways. Metagenomic analysis revealed a shift in microbial community composition, with Massilibacterium enrichment under Fe3O4@AC, highlighting the importance of tailored material design for targeted MCFA production. These findings provide insights into optimizing microbial metabolism for enhanced CE efficiency.