Search Results (777)

Porcine epidemic diarrhea virus (PEDV) has emerged as a major pathogen responsible for porcine diarrheal diseases, causing outbreaks of severe diarrhea and high mortality in neonatal piglets, thereby inflicting severe economic losses on the global swine industry. Current commercial PED vaccines, comprising conventional inactivated and live attenuated formulations, have exhibited progressively diminished efficacy in the face of emerging PEDV variants. The development of high-efficiency vaccine platforms is therefore critical for PED control. This study engineered a cellular membrane nanovesicle (CMN)-based vaccine, which differs from existing inactivated or subunit vaccines by presenting the PEDV spike (S) protein on the cell membranes to mimic the bilayer phospholipid structure of the viral envelope. The full-length S protein (FS, aa 19-1309) or a truncated S protein fragment (TS, aa 19-726) was expressed in Expi293F cells, followed by extraction of cell membranes to assemble antigen-displaying CMN vaccines. Compared with commercial live attenuated vaccine, administration of the CMN vaccine elicited high-titer neutralizing antibodies and elevated IFN-γ-producing CD8⁺ T cells in murine studies. Safety assessments revealed no adverse effects on body weight, hepatic/renal function indices, or histopathological parameters in vaccinated mice. Furthermore, immunization of piglets elicited notable humoral and CD8⁺ T cell immune responses. Collectively, the strategy of CMN-based vaccine described herein delivers a potential PEDV vaccine platform, thereby offering a novel avenue for next-generation veterinary vaccine development. Full article

Pulsed electric fields (PEFs) are widely recognized as a non-thermal, selective physical therapy with wide clinical application in tumor ablation. The pulse width determines how electrical energy is distributed across plasma membrane to intracellular organelles. However, under an engineering-defined equal-dose condition (N·E²·tp), which serves as a practical control parameter rather than a measure of true cellular energy absorption, systematic and comparable experimental characterization of cellular and subcellular responses across pulse widths from the microsecond to nanosecond range remains limited. In this study, PEFs with pulse widths ranging from 100 μs to 50 ns were applied under equal-dose constraints, and cellular responses were evaluated using transmission electron microscopy (TEM), multi-organelle fluorescence imaging, and flow cytometry. The results indicate that pulse-width-dependent effects were observed under a fixed pulse-number, dose-equalized framework in which electric field strength varied across conditions. Structural and functional changes were observed in multiple organelles, including the nucleus, mitochondria, endoplasmic reticulum, and Golgi apparatus. Notably, nanosecond pulses were more effective in inducing mitochondrial membrane potential loss and increasing the proportion of apoptotic or non-viable cells. These findings demonstrate that, under equal-dose conditions, pulse width is a key temporal parameter governing PEF-induced biological effects, indicating that identical dose constraints do not necessarily result in equivalent biological responses. This work provides experimental foundation for parameter selection and optimization in PEF-based biomedical applications. Full article

Background/Objectives: Glioblastoma (GBM) remains the most aggressive primary brain tumor in adults, with limited therapeutic options and poor prognosis despite maximal surgery, radiotherapy, and chemotherapy. The complex and immunosuppressive tumor microenvironment, pronounced intratumoral heterogeneity, and the presence of the blood–brain barrier (BBB) severely restrict the efficacy of conventional and emerging therapies. In this context, cell-based strategies leveraging macrophages, mesenchymal stromal cells (MSCs), and their derivatives have gained attention as “cellular allies” capable of modulating the GBM microenvironment and acting as targeted delivery platforms. Methods: This review systematically analyzes preclinical and early clinical literature on macrophage- and MSC-based therapeutic strategies in GBM, including engineered cells, extracellular vesicles (EVs), membrane-coated nanoparticles, and hybrid biomimetic systems. Studies were selected based on relevance to GBM biology, delivery across or bypass of the BBB, microenvironmental modulation, and translational potential. Evidence from in vitro models, orthotopic and syngeneic in vivo models, and available clinical trials was critically evaluated, with emphasis on efficacy endpoints, biodistribution, safety, and manufacturing considerations. Results: The reviewed evidence demonstrates that macrophages and MSCs can function as active therapeutic agents or delivery vehicles, enabling localized oncolysis, immune reprogramming, stromal and vascular remodeling, and enhanced delivery of viral, genetic, and nanotherapeutic payloads. EVs and membrane-based biomimetic platforms further extend these capabilities while reducing cellular risks. However, therapeutic efficacy is highly context-dependent, influenced by tumor heterogeneity, BBB integrity, delivery route, and microenvironmental dynamics. Clinical translation remains limited, with most approaches at preclinical or early-phase clinical stages. Conclusions: Cell-based and cell-derived platforms represent a promising but still evolving therapeutic paradigm for GBM. Their successful translation will require rigorous biomarker-driven patient selection, improved models that capture invasive GBM biology, scalable GMP-compliant manufacturing, and rational combination strategies to overcome adaptive resistance mechanisms. Full article

The recognition of liquid–liquid phase separation (LLPS) as a widespread organizing principle has revolutionized our view of cellular biochemistry. By forming biomolecular condensates, cells spatially orchestrate reactions without membranes. However, the dysregulation of this precise physical organization is emerging as a driver of diverse pathologies, collectively termed “Condensatopathies.” Unlike traditional proteinopathies defined by static aggregates, these disorders span a dynamic spectrum of material state dysfunctions, from the failure to assemble essential compartments to the formation of aberrant, toxic phases. While research has largely focused on neurodegeneration and cancer, the impact of condensate dysfunction likely extends across broad physiological landscapes. A central unresolved challenge lies in deciphering the “molecular grammar” that governs the transition from functional fluids to pathological solids and, critically, visualizing these transitions in situ. This “material science” perspective presents a profound conundrum for drug discovery: how to target the collective physical state of a protein ensemble rather than a fixed active site. This review navigates the evolving therapeutic horizon, examining the limitations of current pharmacological approaches in addressing the complex “condensatome.” Moving beyond inhibition, we propose that the future of intervention lies in “reverse-engineering” the biophysical codes of phase separation. We discuss how deciphering these principles enables the creation of programmable molecular tools—such as synthetic peptides and state-specific degraders—designed to precisely modulate or dismantle pathological condensates, paving the way for a new era of precision medicine governed by soft matter physics. Full article

A one-dimensional analytical model for a proton exchange membrane fuel cell (PEMFC) engine is presented. The model is structured into three main subsystems: the fuel cell stack, the intake and exhaust system, and the thermal management system. The modeling of the thermal management system specifically encompasses key components such as the expansion tank, thermostat, pump, fan, and radiator. The heat transfer and fluid flow within key thermal management components—primarily fans and radiators—are analyzed via three-dimensional modeling. A porous media model represents the unit parallel-flow radiator, where the complex fin structures are replaced by a homogenized medium. This allows for the efficient calculation of 3D thermal and flow fields once the necessary constitutive parameters are identified. Ultimately, the one-dimensional (1D) thermal management system is coupled with the three-dimensional (3D) flow field analysis. This integrated 1D-3D co-simulation framework is implemented to enhance the computational fidelity of the PEMFC engine’s thermal management model. Full article

Manipulating the mitochondrial genome remains a significant challenge in genetic engineering, primarily due to the mitochondrial double-membrane structure. While recent advances have expanded the genetic toolkit for nuclear and cytoplasmic targets, precise editing of mitochondrial DNA (mtDNA) has remained elusive. Here we report the first successful mitochondrial import of a catalytically active RNA-guided prokaryotic Argonaute protein from the mesophilic bacterium Alteromonas macleodii (AmAgo). By guiding AmAgo to the single-stranded D- or R-loop region of mtDNA using synthetic RNA guides, we observed a nearly threefold reduction in mtDNA copy number in human cell lines. This proof of concept study demonstrates that a bacterial Argonaute can remain active within the mitochondrial environment and influence mtDNA levels. These findings establish a foundational framework for further development of programmable systems for mitochondrial genome manipulation. Full article

Considering the challenges associated with implementing emerging technologies and bacterial-derived antimicrobial metabolites at an industrial scale in the meat industry, this comprehensive review investigates the interactions between lactic acid bacteria-producing antimicrobial metabolites and emerging food preservation technologies applied to meat systems. By integrating evidence from microbiology, food engineering, and molecular physiology, the review characterizes how metabolites-derived compounds exert inhibitory activity through pH modulation, membrane permeabilization, disruption of proton motive force, and interference with cell wall biosynthesis. These biochemical actions are evaluated in parallel with the mechanistic effects of high-pressure processing, pulsed electric fields, cold plasma, irradiation, pulsed light, ultrasound, ohmic heating and nanotechnology. Across the literature, consistent patterns of synergy emerge: many emerging technologies induce structural and metabolic vulnerabilities in microbial cells, thereby amplifying the efficacy of antimicrobial metabolites while enabling reductions in process intensity. The review consolidates these findings to elucidate multi-hurdle strategies capable of improving microbial safety, extending shelf life, and preserving the physicochemical integrity of meat products. Remaining challenges include optimizing combinational parameters, ensuring metabolite stability within complex matrices, and aligning integrated preservation strategies with regulatory and industrial constraints. Full article

Microbial contamination of food is a crucial cause of food spoilage and foodborne diseases, posing a severe threat to global public health. Although chemical preservatives are effective, their potential hazards to human health and the environment, coupled with the growing demand for “clean label” products, have driven the search for natural alternatives. Lactic acid bacteria (LAB), recognized as the Generally Recognized as Safe (GRAS) microorganisms, have emerged as the promising bio-preservatives due to their safety, effectiveness, and multifunctionality. This review systematically summarized the core antimicrobial properties of LAB, including their inhibitory spectrum against foodborne pathogens, spoilage microorganisms, viruses, parasites, and their ability to degrade toxic substances such as mycotoxins, pesticides, and heavy metals. Key inhibitory mechanisms of LAB are highlighted, encompassing the production of antimicrobial metabolites, leading to metabolism disruption and cell membrane damage, nutrition and niche competition, quorum-sensing interference, and anti-biofilm formation. Furthermore, recent advances in LAB applications in preserving various food matrices (meat, dairy products, fruits and vegetables, cereals) are integrated, including their roles in enhancing food sensory quality, extending shelf life, and retaining nutritional value. The review also discusses critical factors influencing LAB’s inhibitory activity (medium composition, culture conditions, ionic components, pathway regulator, etc.) and the challenges associated with the application of LAB. Finally, future research directions are outlined, including the novel LAB and metabolites exploration, AI-driven cultural condition optimization, genetic engineering application, nano-encapsulation and active packaging development, and building up the LAB-based cellular factories. In conclusion, LAB and their antimicrobial metabolites hold great promise as green and safe food preservatives. This review is to provide comprehensive theoretical support for the rational improvement and efficient application of LAB-based natural food preservatives, contributing to the development of a safer and more sustainable food processing and preservation systems. Full article

While drug-eluting cardiovascular devices, including drug-eluting stents and drug-coated balloons, have significantly reduced restenosis rates, they remain limited by delayed vascular healing, chronic inflammation, and late adverse events. These limitations reflect a fundamental mismatch between current device pharmacology, which relies on nonselective antiproliferative drugs, and the highly coordinated, cell-specific programs that orchestrate vascular repair. Extracellular vesicles (EVs), nanometer-scale membrane-bound particles secreted by virtually all cell types, provide a biologically evolved platform for intercellular communication and cargo delivery. In the cardiovascular system, EVs regulate endothelial regeneration, smooth muscle cell phenotype, extracellular matrix remodeling, and macrophage polarization through precisely orchestrated combinations of miRNA, proteins, and lipids. Here, we synthesize mechanistic insights into EV biogenesis, cargo selection, recruitment, and functional effects in vascular healing and inflammation and translate these into a formal framework for EV-inspired device engineering. We discuss how EV-based or EV-mimetic coatings can be designed to sense the local microenvironment, deliver encoded biological “instruction sets,” and function within ECM-mimetic scaffolds to couple local stent healing with systemic tissue repair. Finally, we outline the manufacturing, regulatory, and clinical trial issues that must be addressed for EV-inspired cardiovascular devices to transition from proof of concept to clinical reality. By shifting the focus from pharmacological suppression to biological regulation of healing, EV-based strategies offer a path to resolve the long-standing tradeoff between restenosis prevention and durable vascular healing. Full article

Microbial fuel cells (MFCs) are a promising technology for simultaneously treating wastewater and recovering energy, yet scaling them from lab prototypes to practical systems poses persistent challenges. This review addresses the scale-up gap by systematically examining recent pilot-scale MFC studies from multiple perspectives, including reactor design configurations, materials innovations, treatment performance, energy recovery, and environmental impact. The findings show that pilot MFCs reliably achieve significant chemical oxygen demand (COD) removal (often 50–90%), but power densities remain modest (typically 0.1–10 W m⁻³)—far below levels needed for major energy generation. Key engineering advances have improved performance; modular stacking maintains higher power output, low-cost electrodes and membranes reduce costs (with some efficiency trade-offs), and power-management strategies mitigate issues like cell reversal. Life cycle assessments indicate that while MFC systems can outperform conventional treatment in specific scenarios, overall sustainability gains depend on boosting energy yields and optimizing materials. The findings highlight common trade-offs and emerging strategies. By consolidating recent insights, a roadmap of design principles and research directions to advance MFC technology toward sustainable, energy-positive wastewater treatment was outlined. Full article

Cell membrane-coated nanoparticles represent a biomimetic drug delivery approach that integrates biological membrane functions with synthetic nanomaterials. Among the various membrane sources, those derived from blood cells such as red blood cells, platelets, and leukocytes offer distinctive advantages, including immune evasion, prolonged systemic circulation, and selective tissue targeting. These properties collectively enable efficient and biocompatible delivery of therapeutic agents to diseased tissues, minimizing off-target effects and systemic toxicity. This review focuses on blood cell membrane-derived nanocarriers as drug delivery and immune-regenerative platforms, in which membrane-mediated immunomodulation synergizes with therapeutic payloads to address inflammatory or degenerative pathology. We discuss recent advances in blood cell membrane coating technologies, including membrane isolation, nanoparticle core selection, fabrication techniques, and the development of hybrid and engineered membrane systems that enhance therapeutic efficacy through integrated immune regulation and localized drug action. To illustrate these advances, we also compile membrane type-specific nanocarrier systems, summarizing their core nanoparticle designs, coating strategies, therapeutic cargoes, and associated disease models. Challenges related to biological source variability, scalability, safety, and regulatory standardization remain important considerations for clinical translation. In this review we systematically address these issues and discuss emerging solutions and design strategies aimed at advancing blood cell membrane-based nanocarriers toward clinically viable immune-regenerative therapies. Full article

Exosomes are nanoscale extracellular vesicles that mediate intercellular communication by transporting microRNAs, proteins, and lipids. Generated through Endosomal Sorting Complex Required for Transport (ESCRT)-dependent mechanisms or ESCRT-independent pathways, exosomes are released when multivesicular bodies fuse with the plasma membrane. The ESCRT-dependent pathway involves sequential protein complexes (ESCRT-0, I, II, III) that recognize and sort ubiquitinated cargo, induce membrane budding, and facilitate vesicle scission. In contrast, the ESCRT-independent pathway relies on membrane lipids such as ceramide and proteins like tetraspanins (CD9, CD63, CD81) to promote vesicle formation without ESCRT machinery. Furthermore, post-translational modifications, including ubiquitination, sumoylation, and phosphorylation, further serve as molecular switches, modulating the affinity of ESCRT complexes or cargo proteins for membrane domains and affecting ILV formation rates. In reproductive medicine, exosomes regulate oocyte maturation, embryo–endometrial crosstalk, placental development, and maternal–fetal communication. Altered exosomal signaling contributes to obstetric complications, including preeclampsia, gestational diabetes mellitus, and preterm birth, whereas distinct exosomal miRNA signatures serve as potential diagnostic biomarkers. In gynecology, dysregulated exosomes are implicated in endometriosis, polycystic ovary syndrome, premature ovarian insufficiency, and gynecological malignancies. In contrast, mesenchymal stem cell-derived exosomes show therapeutic promise in restoring ovarian function and enhancing fertility outcomes. The distinctive molecular profiles of circulating exosomes enable minimally invasive diagnosis, while their biocompatibility and ability to cross biological barriers position them as vehicles for targeted drug delivery. Characterization of accessible data provides non-invasive opportunities for disease monitoring. However, clinical translation faces challenges, including standardization of isolation protocols, establishment of reference ranges for biomarkers, and optimization of therapeutic dosing. This review summarizes exosome biogenesis, characterization methods, physiological functions, and clinical applications in obstetrics and gynecology, with an emphasis on their diagnostic and therapeutic potential. Future directions include large-scale biomarker validation studies, engineering approaches to enhance exosome targeting, and integration with precision medicine platforms to advance personalized reproductive healthcare. Full article

Antimicrobial peptides have been increasingly recognized as potential anticancer agents, with the KLA peptide (KLAKLAK₂) being one of the most well-known and successful examples. The research interest in the KLA peptide is attributed to its ability to induce apoptosis in cancer cells by disrupting the mitochondrial membrane. However, the KLA peptide exhibits poor cellular uptake and it lacks targeting specificity, limiting its clinical potential in cancer therapy. In this review, recent advances in nano-engineered delivery platforms for overcoming the limitations of KLA peptides and enhancing their anticancer efficacy are discussed. Specifically, various nanocarrier systems that enable targeted delivery, controlled release and/or improved bioavailability, including pH-responsive nanosystems, photo-chemo combination liposomes, self-assembled peptide-based nanostructures, nanogel-based delivery systems, homing domain-conjugated KLA structures, inorganic-based nanoparticles, and biomimetic nanocarriers, are highlighted. Additionally, synergistic strategies for combining KLA with chemotherapeutic agents or immunotherapeutic agents to overcome resistance mechanisms in cancer cells are examined. Finally, key challenges for the clinical application of these nanotechnologies are summarized and future directions are proposed. Full article

Background: Large-scale production of poultry viral vaccines increasingly requires robust suspension cell platforms. However, most avian cell lines, including DF-1, are strictly anchorage-dependent, limiting scalability. Aquaporin-1 (AQP1) regulates cell–cell adhesion and membrane dynamics, making it a potential target for engineering suspension growth. This study aimed to generate a stable DF-1 suspension cell line via AQP1 disruption and evaluate its potential for enhanced infectious bursal disease virus (IBDV) production. Methodology: DF-1 cells were engineered using a CRISPR/Cas9 ribonucleoprotein system to create a truncated AQP1 gene. DF-1/AQP1⁻ cells were assessed for morphology, tumorigenicity in nude mice, and genetic stability across 20 passages. Suspension growth, cell density, and viability were measured. Cells were infected with IBDV strain BJQ902, and viral titers were compared with wild-type DF-1 and monolayer DF-1/AQP1⁻ cells. Results: DF-1/AQP1⁻ cells maintained normal morphology, were non-tumorigenic, and retained stable AQP1 mutations. They grew as true suspension cultures without adaptation, reaching 4.0 × 10⁶ cells/mL with >95% viability. Suspension DF-1/AQP1⁻ cells cells produced significantly higher viral titers (9.0 log TCID₅₀/mL; 8.63 log EID₅₀/mL) than both monolayer DF-1/AQP1⁻ and wild-type DF-1 cells. Virus production time was shortened in suspension cultures. Conclusions: Targeted AQP1 disruption converts DF-1 cells into a stable, non-tumorigenic suspension cell line with markedly enhanced IBDV production, providing a scalable platform for next-generation avian vaccine manufacturing. Full article

Background/Objectives: Impaired intestinal barrier function is a hallmark of inflammatory bowel disease (IBD). Akkermansia muciniphila and its outer membrane protein Amuc_1100 can enhance this barrier, but the clinical application of Amuc_1100 is limited by the fastidious growth of its native host. This study aimed to overcome this by utilizing the robust probiotic Lacticaseibacillus paracasei L9 for targeted Amuc_1100 delivery. Methods: We engineered Lc. paracasei L9 to express Amuc_1100 via intracellular (pA-L9), secretory (pUA-L9), and surface-display (pUPA-L9) strategies. Their efficacy was assessed in Lipopolysaccharide (LPS)-induced macrophages and a dextran sulfate sodium (DSS)-induced colitis mouse model, evaluating inflammation, barrier integrity, and mucosal repair. Results: The secretory (pUA-L9) and surface-display (pUPA-L9) strains most effectively suppressed pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α) in macrophages. In mice, both strains alleviated colitis and outperformed native A. muciniphila in improving disease activity. Crucially, they exhibited distinct, specialized functions: pUA-L9 acted as a systemic immunomodulator, reducing pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α), elevating anti-inflammatory mediators (IL-4 and IL-10), and promoting goblet cell differentiation; notably, the inhibitory effect of pUA-L9 on IL-6 expression was approximately 2-fold greater than that of pUPA-L9. In contrast, pUPA-L9 excelled in local barrier repair, uniquely restoring mucus layer integrity (Muc1, Muc2, and Tff3) and reinforcing tight junctions (ZO-1, Occludin, Claudin1, Claudin3, and Claudin4). In particular, pUPA-L9 increased Muc2 expression by approximately 3.6-fold compared with pUA-L9. Conclusions: We demonstrate that the subcellular localization of Amuc_1100 within an engineered probiotic dictates its therapeutic mode of action. The complementary effects of secretory and surface-displayed Amuc_1100 offer a novel, spatially targeted strategy for precision microbiome therapy in IBD. Full article