Search Results (696)

Recent studies have highlighted the potential of urinary bladder matrix (UBM) derived from decellularized porcine urinary bladder as a bioactive hydrogel. Despite its complex composition of over 100 proteins, Type I collagen is the primary constituent of UBM. Caco-2 cells are widely used as an in vitro model of the intestinal epithelium; however, to date, no published study has evaluated the effects of UBM on Caco-2 cells. In this study, Electric Cell–Substrate Impedance Sensing (ECIS) was used to measure Caco-2 cell attachment and wound-healing migration on UBM-coated microelectrodes. Our results demonstrate that UBM hydrogel coating at 0.2 mg/mL significantly accelerates cell attachment and enhances migration rates compared to uncoated controls. These stimulatory effects were comparable to those observed with 0.2 mg/mL Type I collagen, suggesting that UBM can function as effectively as Type I collagen. We further monitored barrier formation in Caco-2 cells cultured on UBM-coated transwell membrane inserts using TEER measurements and scanning electron microscopy. The TEER values reached 300 Ω·cm² within three days, indicating the rapid establishment of mature tight junctions. Overall, these results show that UBM hydrogel coatings are effective substrates for Caco-2 cells, performing as well as Type I collagen in all our tests. Full article

Background/Objectives: Self-amplifying RNA (saRNA) vectors enable high-level transgene expression from minimal initial doses. While alphavirus-based saRNA systems are widely used, they suffer from limitations, including large genome size, complex replicase machinery, and cellular toxicity. Nodamura virus (NoV) offers a promising alternative due to its compact genome (3.2 kb) and low cytotoxicity. This study aimed to elucidate NoV RNA1 replication mechanisms and develop a novel NoV-based saRNA vector platform. Methods: We established a Schizosaccharomyces pombe system to investigate NoV RNA1 replication and protein A localization. N-terminal deletion mutants and ER-targeting chimeras were constructed to characterize membrane targeting determinants. Based on mechanistic insights, we developed NovaVec by inserting transgenes at the RNA3⁴²² site within the subgenomic RNA3 region. In vivo performance was evaluated using lipid nanoparticle-encapsulated NovaVec expressing nanoluciferase or monkeypox A33R antigen in BALB/c mice. Results: We identified redundant mitochondrial targeting domains (amino acids 2-15 and 16-33) in NoV protein A, where either domain was sufficient for proper localization and replication. The replication machinery could be functionally redirected to the endoplasmic reticulum while maintaining replication competence. Lipid nanoparticle-encapsulated NovaVec achieved sustained transgene expression for 54 days in mice, significantly outperforming conventional mRNA vectors that lost signal within 14 days. The NovaVec-based monkeypox A33R vaccine elicited robust antigen-specific humoral immunity with titers reaching approximately 1:12,800 following booster immunization. Conclusions: With its compact genome encoding only a single replicase protein, minimal cytopathic effects, and demonstrated capacity for long-term protein expression, NovaVec represents a highly promising next-generation saRNA platform for vaccines. Full article

The Kelch-repeat superfamily genes played important roles in regulating plant growth and development; however, their functions in foxtail millet (Setaria italica) have not yet been characterized. In this study, SiNL4, a homolog of ZmNL4 controlling leaf width in maize, was knocked out using the CRISPR/Cas9 technology, and two homozygous knockout lines (ko1 and ko2) were obtained. Phenotypic analysis showed that compared with the wild-type Ci846, ko1 and ko2 exhibited reduced leaf width and decreased yield related traits (e.g., panicle weight, grain width, and 1000-grain weight). Cytological analysis showed that changes in leaf width of ko1 and ko2 resulted from a decrease in leaf epidermal cell width and the number of small vascular bundles (SVBs) close to the leaf edge, suggesting that SiNL4 might regulate leaf width by influencing cell expansion and the development of SVB. Spatiotemporal expression analysis indicated that the relative expression level of SiNL4 was high in the stem, leaf, and young panicle. Subcellular localization showed that SiNL4 was mainly localized in the mitochondria and plasma membrane. In addition, the T-DNA insertion mutant (Atnl4) of AT5G18590, the ortholog of SiNL4 in Arabidopsis thaliana, exhibited similar phenotypes with reduced rosette leaf width, seed width, and 1000-seed weight. Moreover, complementary expression of SiNL4 in Atnl4 not only restored the phenotypes, but also significantly increased the 1000-seed weight, indicating that the function of these two genes might be conserved. Meanwhile, we found that SiNL4 knockout caused a decrease in chlorophyll content and net photosynthetic rate (Pn), showing that SiNL4 might be involved in regulating photosynthesis. In summary, this study revealed the function of SiNL4 in regulating leaf width in foxtail millet, providing a potential gene for the genetic improvement of foxtail millet. Full article

Background/Objectives: Antimicrobial peptides (AMPs) are promising candidates against multidrug-resistant bacteria, although their clinical translation is frequently limited by cytotoxicity. In this study, we investigated whether combinations of structurally related oligomeric analogs could cooperatively enhance bacterial membrane targeting while redistributing the associated cytotoxic burden. Methods: Monomeric, dimeric, and tetrameric AMPs were evaluated through antimicrobial susceptibility testing, checkerboard interaction assays, RAW 264.7 macrophage cytotoxicity assays, and all-atom molecular dynamics simulations, including biased membrane insertion and umbrella sampling analyses. In addition, we introduced the Combinatorial Therapeutic Index (CTI) as an exploratory metric to estimate the predicted reduction in cytotoxic burden associated with peptide combinations. Results: Cytotoxicity varied substantially among oligomeric forms, with larger and more hydrophobic peptides, particularly tetramers, exhibiting the highest cytotoxicity. Additive effects were observed in checkerboard assays involving linear, dimeric, and tetrameric forms, supporting the redistribution of the toxic burden and enabling the beneficial membrane-interaction properties of hydrophobic linear peptides to be leveraged at lower cytotoxic cost. Predicted therapeutic improvement ranged from approximately twofold for the SET-M33:L33 combination to nearly ninefold for the SET-M33:DIM-33:L8 triple combination. Molecular dynamics simulations revealed non-redundant membrane interaction behaviors, with smaller peptides exhibiting deeper membrane insertion and the dimeric form favoring interfacial membrane engagement. Conclusions: These findings support a cooperative formulation strategy in which structurally related SET-M33 oligomers contribute complementary antibacterial functions while reducing the predicted cytotoxic burden. Further experimental validation using direct cytotoxicity assays of complete peptide mixtures will be necessary to confirm the therapeutic potential of these formulations. Full article

Background: The socket preservation technique involves filling the bone defect that occurs after a tooth is extracted with bone substitute material. This procedure helps to minimize bone resorption of the alveolar ridge following extraction. Various bone substitute biomaterials can be used for augmentation, including autogenous, allogeneic, and xenogeneic options. This study aimed to assess changes in alveolar ridge dimensions and variations in radiographic bone density in sockets grafted with two distinct biomaterials. Furthermore, bone biopsies collected from the grafted sites were subjected to histological analysis. Methods: Forty generally healthy patients were enrolled in the study and split into four equal groups. The first and third groups underwent first or second maxillary premolar extraction and received treatment with an allogeneic material (BIOBank®, Biobank, Paris, France), while the second and fourth groups underwent first or second mandibular molar extraction and were treated with a xenogeneic material (Geistlich Bio-Oss®, Geistlich Pharma AG, Wolhusen, Switzerland). Following tooth extraction, the appropriate biomaterial was inserted into the socket. It was covered with a collagen membrane (Geistlich Bio-Gide®, Geistlich Pharma AG, Wolhusen, Switzerland) and stabilized with sutures, which were removed seven to ten days after the procedure. Micro-CBCT scans were conducted to evaluate the dimensions of the alveolar ridge and the radiographic density of the grafted socket at 7–10 days and 6 months after the procedure. A bone trepanobiopsy was performed concurrently with implant placement six months after socket preservation. The obtained biopsy was analyzed histologically using hematoxylin and eosin (H&E) and Masson’s trichrome staining. Results: There was no statistically significant difference in alveolar ridge height or width preservation between allogeneic and xenogeneic biomaterials. After six months of healing, sockets grafted with both materials exhibited greater radiographic bone density, with significantly greater density observed in the xenograft group. Conclusions: The findings of this study suggest that the two biomaterials are comparable in their effectiveness at maintaining the dimensions of the alveolar ridge. However, the quality of the newly formed bone may differ depending on the type of biomaterial used. Full article

Background/Objectives: BRI is a synthetic arylamide polymer designed to mimic the electrostatic and amphiphilic features of defensin-type antimicrobial peptides (AMPs), although its molecular organization and activity have not been experimentally validated. This study presents the first integrated computational and experimental characterization of BRI to define the physicochemical basis of its AMP-like behavior and membrane interactions. Methods: Molecular modelling was used to evaluate the structural and electrostatic properties of BRI. Model lipid membranes were used to study membrane interactions. Fluorescence spectroscopy, electrochemical measurements, and ζ-potential analyses were performed to characterize membrane insertion, aggregation, ionic conductance, and membrane resistance. Microbiology assays evaluating synergy with azole were also assessed. Results: Molecular modelling showed that BRI is a flexible molecule with cationic and hydrophobic surfaces, a strong amphiphilic dipole, and a dominant +4 charge state, explaining its sensitivity to ionic strength and membrane interactions. BRI displayed two membrane-dependent mechanisms of action. In zwitterionic phospholipid membranes, BRI resembled canonical AMPs, showing membrane insertion, pore formation, and increased ionic conductance. In anionic ergosterol-containing membranes mimicking fungal cells, BRI exhibited sterol-dependent insertion, in-plane aggregation, and modulation of membrane resistance without pore formation. Fluorescence, electrochemical, and ζ-potential measurements supported BRI–BRI interactions at the membrane interface and sensitivity to lipid domain organization. BRI also synergized with azole antifungal drugs, suggesting a mechanistic role for ergosterol in its antifungal activity. Conclusions: These findings reveal a sterol- and domain-mediated mechanism for arylamide polymers and identify lipid organization as a key determinant of antifungal activity. The dependence of BRI activity on ergosterol content provides a mechanistic explanation for its synergy with azole antifungals and supports further investigation of BRI as a membrane-active antifungal agent. Full article

One-dimensional mass transfer resistance-in-series framework was developed theoretically and validated experimentally using a flat-plate polytetrafluoroethylene/polypropylene (PTFE/PP) membrane module to predict CO₂ absorption fluxes and concentration distributions. The decline in CO₂ absorption efficiency along the membrane module is primarily attributed to increased concentration polarization resistance and a reduced driving force concentration gradient. To alleviate these limitations, carbon fiber promoters were strategically embedded to suppress concentration polarization, reduce the mass transfer resistances, and enhance turbulence intensity. In the present study, device performance was further improved by implementing properly ascending or descending hydraulic equivalent widths along the absorbent feed channel. Under the descending configuration, an absorption flux enhancement of up to 44.94% was achieved relative to an empty-channel module (i.e., without S-ribbed carbon fiber inserts). Theoretical formulations were established to predict absorption fluxes under varying monoethanolamine (MEA) volumetric flow rates, CO₂/N₂ mixture flow rates, and inlet CO₂ feed concentrations. The model predictions showed good agreement with experimental results obtained using MEA solutions under both ascending and descending hydraulic width operations, demonstrating effective mitigation of polarization effects and enhanced absorption flux along the absorbent feed channel. An economic assessment of the S-ribbed carbon fiber module was conducted by evaluating the trade-off between absorption flux enhancement and incremental power consumption. The results indicate that the proposed design provides a practical and economically viable approach for improving the performance of membrane-based CO₂ capture technologies. In addition, an enhanced Sherwood number correlation, expressed in a simplified form, was developed and employed to estimate the mass transfer coefficients of CO₂ membrane absorption modules incorporating S-ribbed carbon fiber promoters. Full article

Background/Objectives: Longer oligoarginines are very effective cell-penetrating peptides. It has been shown that a minimal number of positively charged side chains is necessary for efficient cellular uptake. But a highly positively charged peptide may interact with its cargo molecule, thereby reducing its efficiency. Several chemical modifications were tested to improve the internalization of short tetraarginine derivatives. Aromatic groups, such as Dabcyl at the N-terminus, Trp in the sequence, and AMBA or PABA in the backbone, were used to improve internalization. The other useful modification was the aza-glycine substitution in the case of penetratin. Methods: In this study, the effect of aza-glycine insertion into the peptide Dabcyl-RRRRK(Cf) on internalization was studied and compared with that of the Trp-modified peptide Dabcyl-RRWRRK(Cf). To explain the noticed difference in the biological activity of peptides, DFT calculations and the prediction of membrane-binding free energy (ΔΔF) from a peptide sequence were performed. Results: It turned out that the position of the aza-glycine moiety does not have an influence on the cellular uptake. The aza-glycine-containing peptide showed higher internalization than the Dabcyl-RRRRK(Cf) peptide. Besides this, these peptides have similar or higher cellular uptake than that of octaarginine at lower concentrations (c < 2 µM). The aza-glycine affected not only cellular uptake but also the entry mechanism. The structure of peptides depended on the amino acids (Trp, Gly, or azaGly) in their sequences and their positions. Conclusions: These may result in the different amphiphilicity of peptides, and thus changes in the hydrophobic moment and in the binding affinity of peptides to the negatively charged membrane surface. Full article

Natural killer (NK) cells are promising effectors for cancer immunotherapy, as they can recognize and eliminate tumor cells without prior antigen sensitization. However, insufficient tumor recognition remains a critical limitation that reduces the anticancer efficacy of NK cells against solid tumors. To address this limitation, we developed a lipid-mediated cell membrane engineering strategy to enhance the targeting and cytotoxic efficacy of NK cells toward solid tumors, particularly ovarian cancer cells. In this strategy, poly-L-glutamic acid (PLE) was employed as an ovarian cancer-targeting module due to the specific affinity of PLE for cholesterol-rich membrane domains. To display PLE on NK cells, a lipid moiety is incorporated to anchor PLE onto the NK cell membrane via hydrophobic insertion, enabling rapid and non-genetic surface modification. As a result, the surface-engineered NK cells with PLE-Lipid (i.e., PLE-NK) displayed PLE on the NK cell surface, allowing direct recognition of ovarian cancer cells without compromising the intrinsic properties of NK cells. This enhanced recognition subsequently increased NK–cancer cluster formation by promoting interactions between membrane-presented PLE on NK cells and cholesterol on ovarian cancer cells. Consequently, PLE-NK cells exhibited enhanced cytotoxicity against ovarian cancer cells (i.e., OVCAR-3 cells) and effectively disrupted 3D tumoroids, while PLE-NK cells showed no off-target effects on normal fibroblasts. Collectively, these findings demonstrate that PLE-Lipid-mediated NK surface engineering provides a simple and effective strategy to improve the tumor targeting ability of NK cells and offers a promising platform for NK cell-based immunotherapy against ovarian cancer. Full article

Background: The reliable co-loading of paclitaxel (PTX) and indocyanine green (ICG) into a single lipid nanoparticle (LNP) enables synergistic antitumor delivery but remains challenging due to their distinct physicochemical properties. Methods: This study integrated COSMO-RS calculations, molecular dynamics simulations, and in vitro assays to systematically investigate the effects of lipid composition, drug modification, particle size, and solvent environment on dual-drug loading. Results: This work indicate that DMPS lipid membranes featuring highly polar headgroups and ordered bilayer structures stably bind both ICG and PTX, achieving drug-loading efficiencies (DLEs) of 7.2% and 5.6%, respectively. Carboxylation of PTX enhanced hydrogen bonding with DMPS, while alkyl chain modifications improved membrane insertion, though excessive chain length (e.g., C12) reduced stability due to increased flexibility. Increasing the LNP size from 50 nm to 250 nm raised the DLE of PTX from 4.7% to 8.1%, while sizes beyond 500 nm led to membrane destabilization. The use of 20 vol% ethanol increased total drug loading by 51% by disrupting the hydration shell of ICG and suppressing PTX aggregation; however, ethanol concentrations exceeding 40 vol% intensified drug–solvent competition and weakened membrane binding. Conclusions: This study provides a comprehensive elucidation of the multifactorial regulatory mechanisms underlying dual-drug loading in LNPs, offering a theoretical basis for the rational design of efficient co-delivery systems. Full article

Bacterial β-barrel pore-forming toxins, including Staphylococcus aureus α-toxin (Hla) and Bacillus cereus toxins hemolysin II (HlyII) and cytolytic toxin K2 (CytK-2), are secreted by bacterial cells as water-soluble monomers. These monomers assemble within lipid bilayers to form cylindrical pores, leading to lysis of target eukaryotic cells. We created mutant forms of these toxins that, based on the results of X-ray structural analysis of Hla and the prediction of the 3D structure of HlyII and CytK2, can form intramolecular disulfide bonds in monomers. The substitutions were made in the region responsible for toxin insertion into the target membrane. The mutant forms reversibly altered their hemolytic activity depending on the presence of reducing reagents and were non-toxic when injected into experimental animals. The immune response to injection of the mutant forms of Hla and CytK-2 toxins resulted in higher antibody titers against the wild-type toxins and a higher level of immunological memory than with injection of the HlyII mutant. The mutant form of CytK-2 demonstrates the properties of a prototype vaccine, as immunization with this protein protects animals against the effects of the wild-type toxin. Full article

Physical integration between endosymbiotic algae and host mitochondria is a recurring feature across photosynthetic symbioses, yet the structural nature of this association has remained unresolved. In the ciliate Paramecium bursaria, each endosymbiotic Chlorella cell is enclosed by a perialgal vacuole (PV) membrane consistently surrounded by host mitochondria, suggesting a conserved architecture for metabolic interaction. Although transmission electron microscopy has shown close membrane apposition, it has remained unclear whether this reflects incidental proximity or a reinforced adhesion. Here, we provide direct evidence that the PV membrane and host mitochondrial membrane form a stable physical association. Using discontinuous Percoll centrifugation, we isolated intact units in which Chlorella and mitochondria co-sedimented, indicating that their association withstands mechanical disruption. By fluorescently labeling the PV and mitochondrial membranes with BODIPY FL C₅-ceramide (BC₅C), together with a mitochondria-specific monoclonal antibody and DAPI, we visualized the PV membrane under light microscopy and demonstrated that the mitochondrial–PV membrane complex persists after homogenization and centrifugation. As expected from the membrane-insertion behavior of BC₅C, this fluorescent labeling revealed that the PV–mitochondrial membrane association is structurally reinforced rather than incidental, providing a mechanistic framework for understanding how Chlorella cells are stably positioned beneath the host cortex. Full article

Background/Objectives: Fungal keratitis remains a vision-threatening infection, and current amphotericin B (AmphB) eye drops suffer from low corneal residence time, poor aqueous solubility, and the need for frequent dosing. This study develops electrospun nanofiber-based ophthalmic inserts combining polyvinyl alcohol (PVA), gamma-cyclodextrin (γ-CD), and sodium taurocholate (STC) to enhance AmphB solubility and provide a non-invasive, rapidly dissolving ophthalmic dosage form. Methods: γ-CD and STC-enhanced AmphB-loaded PVA nanofiber-based ophthalmic inserts with varying γ-CD and STC concentrations were prepared by electrospinning and characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD). Drug content, in vitro release (Weibull modeling), antifungal activity against Candida albicans, Fusarium solani, and Aspergillus fumigatus, ocular cytocompatibility using the Hen’s Egg Test on Chorioallantoic Membrane (HET-CAM), and accelerated stability (40 ± 2 °C, 75 ± 5% relative humidity, 4 weeks) were evaluated. Results: Bead-free nanofibers with mean diameters between 216 ± 33 nm and 310 ± 35 nm were obtained, and XRD confirmed complete amorphization of AmphB within the PVA nanofiber matrix, forming an amorphous solid dispersion. All formulations showed rapid and nearly complete AmphB release (≈100% within 60 min), with Weibull β values < 0.75, indicating Fickian diffusion-controlled release. AmphB-loaded PVA nanofiber-based ophthalmic inserts produced inhibition zones and broth susceptibility profiles comparable to AmphB in dimethyl sulfoxide (DMSO), demonstrating preserved antifungal activity. HET-CAM scores (0–0.9) classified the inserts as practically non-irritant, and SEM/FTIR after accelerated storage showed no relevant morphological or physicochemical changes. Conclusions: These γ-CD and STC-enhanced AmphB-loaded PVA nanofiber-based ophthalmic inserts provide a non-invasive, rapidly dissolving ophthalmic dosage form that combines amorphous AmphB, immediate drug availability, and good ocular tolerance, supporting their further development as a patient-friendly treatment option for fungal keratitis. Full article

The potential of water-soluble membrane proteins (wsMPs) has not been fully realized. In this article, we exploit the nearly identical functionality of wsMPs with their membrane-bound counterparts and show that we can create water-soluble membrane proteins that incorporate into the plasma membranes of cells and alter their fate. As a proof of concept, we demonstrate the functional properties of water-soluble engineered pore-forming proteins, K⁺ ionic channels (MthK), and constitutively active GPCRs—among them frizzled receptors—both in vitro and in vivo. We call this method in vivo deployment of recombinant viable MPs, iDRIVE. Furthermore, we demonstrate that our strategy mediates the unidirectional insertion of MPs into the plasma membrane, and through constitutively active receptors, we present evidence for similar signaling pathway activation between small molecules and our water-soluble proteins using model phenotypes and molecular signaling assays. We present three examples where wsMPs are functional in dictating cellular fate, both in vitro and in vivo. Lastly, we show the induction of similar differential methylation via the activation of the Wnt signaling pathway using the conventional small molecule agonist, CHIR99021, or our wsFrizzled receptors (iDRIVE-FZD) in human embryonic kidney (HEK 293) embryoid spheroids (ESs). Additionally, we show that Wnt activation via wsFrizzled receptors results in even more biologically relevant epigenetic changes than via the small molecule CHIR99021. Future work will employ iDRIVE to differentiate stem cells in the production of research and clinically relevant organoids. Full article

Background: Rabies is a highly fatal zoonotic disease that causes approximately 59,000 human deaths worldwide each year. Current inactivated rabies vaccines require multiple doses and are associated with high costs. The full-length rabies virus glycoprotein (RVG), a membrane protein, exhibits substantial instability in its trimeric structure during recombinant expression. This instability makes it difficult to obtain high-purity, correctly folded antigens. Objectives: This study focuses on the preparation of a full-length recombinant RVG subunit vaccine candidate expressed in a glycoengineered Pichia pastoris system with mammalian-like glycosylation. Methods: The full-length RVG gene (including the transmembrane domain and cytoplasmic tail) from the Challenge Virus Standard-11 (CVS-11) strain was codon-optimized and inserted into the pPICZαA vector to construct the recombinant expression plasmid pPICZαA-RVG. The plasmid was transformed into glycoengineered Pichia pastoris X33-7 (low-mannose type) by electroporation for inducible expression. The target protein was purified by nickel affinity chromatography, anion-exchange chromatography, and Superdex-200 size-exclusion chromatography. The structural characteristics of the purified protein were analyzed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The purified antigen was formulated with the adjuvants AS03 or MF59. BALB/c mice (n = 5 per group) were immunized intramuscularly following a four-dose schedule (days 0, 7, 14, and 28). Antigen-specific IgG antibody titers were measured by ELISA, and neutralizing antibody titers were determined using the rapid fluorescent focus inhibition test (RFFIT). Results: Glycoengineered Pichia pastoris yeast strains expressing wild-type RVG (RVG-WT) or a mutant variant (RVG-M6: R84S, R199S, H270P, R279S, K300S, and R463S) were successfully constructed. The purified RVG antigen formed nanoparticles with an average particle size of approximately 75 nm. Immunized mice generated robust RVG-specific IgG responses, with titers reaching approximately 6.31 × 10⁵ for RVG-WT after the fourth immunization, compared to 3.16 × 10³ for RVG-M6 and 5.62 × 10³ for the RVG-WT-PEG control. Two weeks after the fourth immunization, RVG-WT formulated with AS03 or MF59 induced significant neutralizing antibody responses compared with the control group (p < 0.0001 and p < 0.01, respectively). The neutralizing antibody titers reached 1:79.43 in the AS03 group and 1:33.11 in the MF59 group, whereas the WT-PEG + AS03 control group showed a low titer of 1:3.72. In contrast, RVG-M6 formulated with MF59 failed to induce detectable neutralizing antibodies (1:3.02). Furthermore, RVG-WT + AS03 induced significantly higher neutralizing antibody responses than the WT-PEG + AS03 control group (p < 0.0001), and a significant difference was also observed between the RVG-WT + MF59 and RVG-M6 + MF59 groups (p < 0.01). Conclusions: The glycoengineered Pichia pastoris expression system successfully produced uniform full-length rabies virus glycoprotein nanoparticles with high purity. When formulated with the AS03 adjuvant, RVG-WT induced high-titer neutralizing antibodies in mice, suggesting a promising strategy for the development of recombinant subunit vaccines against rabies. However, this study is limited by the absence of challenge studies and validation in target animal species, which will be further investigated in future work. Full article