Search Results (4,002)

Sentiment analysis of online game reviews has increasingly become important in understanding player experiences and supporting data-driven game development. However, research in this domain has continuously faced two unresolved challenges: (1) the extreme imbalance between positive and negative feedback, and (2) the inefficiency of existing feature-weighting schemes in capturing sentiment signals embedded in informal gaming discourses. Prior works demonstrated that negative feedback—though a few in number are highly influential—usually contain richer emotional content and longer textual structures; yet, prevailing classification models often perform poorly for these minorities (i.e., negative feedback). Numerous studies explored multimodal imbalance issues, class imbalance in cross-lingual ABSA (Aspect-Based Sentiment Analysis), reinforcement-learning-based architectures for imbalanced extraction tasks, and oversampling strategies like SMOTE (Synthetic Minority Over-sampling Technique) variants. Few investigations specifically addressed imbalanced sentiment classification in the contexts of online game reviews, where user-generated content exhibits unique lexical, structural, and emotional characteristics. To address these gaps, this study integrated TF-IDF (Term Frequency-Inverse Document Frequency), VADER (Valence Aware Dictionary and Sentiment Reasoner) lexicon features, and IGM (Inverse Gravity Moment) weightings with advanced oversampling methods such as ADASYN (Adaptive Synthetic Sampling Approach for Imbalanced Learning) and Borderline-SMOTE to improve the detection of minority sentiment classes. Ensemble models, including XGBoost (Extreme Gradient Boosting) and LightGBM (Light Gradient-Boosting Machine), were further employed to enhance the robustness of imbalance. Using a large-scale dataset of Steam game reviews, the proposed framework demonstrated substantial improvement in identifying negative sentiments, addressing a critical limitation in the existing computational game-analysis literature, and advancing the modeling for detecting the emotion-rich but imbalance-prone user feedback. Full article

The environmental impacts from fossil fuel use have accelerated the global transition to sustainable energy sources. Hydrogen has become a promising alternative due to its high energy density and clean combustion. However, hydrogen production streams are frequently contaminated with methane, which needs efficient, durable, and cost-effective purification technologies such as pressure swing adsorption (PSA). The present study provides a comparative evaluation of biomass-derived activated carbons and a commercial activated carbon for hydrogen–methane separation. High-surface-area activated carbons were synthesized from sustainable pine and birch precursors via chemical activation using potassium hydroxide (KOH, impregnation ratio 3:1) at 800 °C. Their dynamic adsorption performance was systematically assessed in a fixed-bed setup under a PSA system operating at pressures of 25, 35, and 50 bar, using a of hydrogen–methane gas mixture, where methane feed concentrations ranging from 10 to 30 vol%. This work focuses on the behavior of the adsorbent material and does not constitute a complete PSA process evaluation. The biomass-derived activated carbons showed well-developed textural characteristics, with specific surface areas up to 1416 m² g⁻¹, which exceeded that of the commercial reference material (1023 m² g⁻¹). This improved pore structure was reflected in their adsorption behavior at an operating pressure of 50 bar; the birch-derived carbon achieved a methane uptake of 10.5 mol kg⁻¹, more than twice the capacity of 5.30 mol kg⁻¹ measured for the commercial adsorbent. Beyond initial adsorption capacity, the study emphasizes operational durability and reusability. Cyclic adsorption–desorption experiments, supported by Raman spectroscopy, revealed pronounced structural changes in the commercial activated carbon under repeated operational stress, as indicated by an increase in the I_D/I_G ratio from 1.08 to 1.24. In contrast, the biomass-derived activated carbons preserved their morphological integrity and adsorption efficiency over successive cycles. These findings demonstrate that pine- and birch-derived activated carbons are not only sustainable alternatives but also operationally stable adsorbents capable for hydrogen purification processes. Full article

Background: Pneumococcal conjugate vaccines (PCVs) prevent severe disease in children, but high costs limit access. PCV10-SII (PNEUMOSIL), a 10-valent PCV prequalified by the World Health Organization (WHO) in 2019, offers a cost-effective alternative. This study assessed its safety and immunogenicity in Vietnamese children aged 6 weeks–24 months. Methods: An open-label, single-arm study enrolled 304 children in three age groups: 6 weeks–6 months (n = 151), >6–12 months (n = 76), and >12–24 months (n = 77). Participants received two or three doses. Safety was evaluated through immediate reactions, adverse events (AEs), serious adverse events (SAEs), and withdrawals. Immunogenicity was measured 28 days after the final dose using serotype-specific IgG geometric mean concentrations (GMCs), opsonophagocytic activity (OPA) titers, and seroresponse rates. The trial was approved by the IRB of the National Ethics Council (code: No. 75/CN-HĐĐĐ on date 4 June 2021) and was registered with ClinicalTrials.gov, NCT05140720. Results: Of 304 enrolled participants, 294 (96.7%) completed follow-up. No immediate adverse events or serious adverse events occurred. Unsolicited adverse events were reported in 17%, mainly respiratory, while serious adverse events occurred in 4%. Mild local/systemic reactions (e.g., injection site pain, crying) resolved without sequelae. Immunogenicity was strong, with GMCs 1.8–9.11 µg/mL, GMTs 277.8–22,342, and seroresponse rates >90% for 9 of 10 serotypes, serotype 6B demonstrated a slightly lower seroresponse rate of 88.6%. Conclusions: PCV10-SII demonstrated favorable safety and robust immunogenicity, supporting its inclusion in national immunization programs as an affordable option for pneumococcal disease prevention. Full article

(1) Background: Potentially arthropod-pathogenic and plant-associated Metarhizium fungi are of high interest for basic research, biological pest control and plant growth promotion. Unambiguous species delineation enabling the taxonomic assignment of new isolates and the identification of new Metarhizium species is of crucial importance for both research and application. Recently, the new species Metarhizium hybridum and Metarhizium parapingshaense were introduced on the basis of phylogenomic studies. (2) Methods: Neighbor- joining and Bayesian inference-based phylogenetic reconstruction of ribosomal intergenic spacer (rIGS) sequences were used to critically evaluate new species introductions. A species-discriminating diagnostic PCR tool for Metarhizium was adapted to M. hybridum and M. parapingshaense. GenBank database mining was performed to identify cryptic descriptions of the new species. (3) Results: The introduction of M. hybridum and M. parapingshaense was corroborated by rIGS sequence comparison. Data mining revealed cryptic first descriptions of M. hybridum from Canada, China, Colombia, Costa Rica, Cuba, Honduras, Mexico, New Zealand, the USA and the Philippines, and of M. parapingshaense from China, India, Japan, the Philippines and South Korea. (4) Conclusions: Results support the reliability of rIGS as a single taxonomic marker for species-level identification of Metarhizium fungi. Species-discriminating diagnostic PCR was successfully adapted to enable the sequencing-independent identification of the confirmed new species M. hybridum and M. parapingshaense. Full article

Biosensors based on electrolyte-gated organic field-effect transistors (EGOFETs) have attracted considerable attention due to their advantages, including low cost, inherent signal amplification, and low-voltage operation. A critical step influencing sensing performance is the integration of specific receptors onto the device surface. Among various strategies, the covalent immobilization of biorecognition elements onto gold surfaces via thiol chemistry is one of the most widely used approaches. In this study, we report the optimization of a mixed self-assembled monolayer (SAM) composed of 11-mercaptoundecanoic acid (11-MUA) and 3-mercaptopropionic acid (3-MPA) for label-free detection of human IgG using EGOFETs. The quality of the SAM was systematically modulated by varying the total concentration from 10 to 400 mM and characterized using X-ray Photoelectron Spectroscopy (XPS), Electrochemical Impedance Spectroscopy (EIS), Cyclic Voltammetry (CV), and Atomic Force Microscopy (AFM). The results revealed that a concentration of 50 mM yielded a densely packed and well-ordered monolayer. After covalent immobilization of anti-IgG antibodies via 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) chemistry and subsequent blocking with ethanolamine and bovine serum albumin (BSA), the functionalized gate electrodes were integrated into poly(3-hexylthiophene) (P3HT)-based EGOFETs. Electrical measurements demonstrated that EGOFET biosensors functionalized with the 50 mM SAM achieved optimal sensing performance. The devices exhibited a highly linear response (R² = 0.998) over a wide concentration range from 1 fM to 10 nM, with a LOD of 2.82 fM, and showed excellent selectivity against non-target immunoglobulins A and M (IgA and IgM). This SAM concentration optimization strategy provides a versatile approach for engineering high-performance EGOFET biosensors, with potential applicability to a broad range of disease biomarkers. Full article

The incidence of furunculosis in juvenile Atlantic salmon, Salmo salar, has increased in recent years in Chile, with isolates of Aeromonas salmonicida being the primary cause. However, in some cases, molecular diagnostics failed to identify the etiological agent. We previously demonstrated that a proportion of undiagnosed cases was produced by a new A. salmonicida strain. In those cases where the pathogen remained unidentified, we isolated colonies with an A. salmonicida-like appearance. Subsequent phylogenetic analysis presented in this work grouped those A. salmonicida-like isolates within the Aeromonas piscicola clade. Whole genome sequencing confirmed the taxonomic affiliation, giving additional insights into virulence and antibiotic resistance markers. Indeed, one of the strains showed reduced susceptibility to oxytetracycline. Virulence potential was assessed by in vivo testing in S. salar, which resulted in disease with pathognomonic signs of furunculosis. Although the pathogen presents common antigens with A. salmonicida, the current vaccine triggered only a modest IgM response against A. piscicola in the field. Our results support the hypothesis that the increasing incidence of furunculosis in Chile cannot solely be ascribed to the emergence of the new less-virulent A. salmonicida strain, but may partially result from furunculosis-like infections caused by A. piscicola strains which exhibit a comparable virulence level. 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

This study employed UPLC-MS/MS to determine the contents of major polyphenolic compounds and proanthocyanidins (PCs) in Kyoho grape seeds, optimized the extraction method and conditions for PCs using response surface methodology (RSM), and further evaluated the scavenging activities of PCs against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl (•OH) radicals as well as their effects on growth, immunity, and oxidative stress in mice. Three hundred and sixty 3-week-old male mice (42.28 ± 0.31 g) were assigned to a single factor complete randomized trial design and fed with six different diets including 0 mg/kg vitamin E(VE) + 0 mg/kg PCs, 100 mg/kg VE, 25 mg/kg PCs + 75 mg/kg VE, 50 mg/kg PCs + 50 mg/kg VE, 75 mg/kg PCs + 25 mg/kg VE and 100 mg/kg PCs, respectively. The results demonstrated that PCs were identified as the predominant phenolic compounds, accounting for 29.6% of total phenolic substances in Kyoho grape seeds. Additionally, the ultrasound-assisted extraction method was superior to the shaker-assisted and low-temperature infiltration extraction methods, with optimal conditions of 60% ethanol concentration, material-to-liquid ratio of 1:20 g/mL, temperature of 30 °C, and extraction time of 50 min. Scanning electron microscopy (SEM) revealed that ultrasound treatment effectively disrupted the seed surface structure, facilitating PC release. In vitro, PCs exhibited significantly stronger DPPH and hydroxyl radical (•OH) scavenging activities than vitamin C (VC), Trolox, and gallic acid. Compared with the control group, mice fed diets containing PCs and VE showed higher superoxide dismutase (SOD) activity, glutathione peroxidase (GSH-PX) activity, and total antioxidant capacity (TAOC), Catalase (CAT), GPX and inflammation factor 10 (IL-10) genes levels in the serum and liver (p < 0.05), whereas the levels of immunoglobulin G (IgG), immunoglobulin A (IgA), immunoglobulin M (IgM), tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6), as well as the mRNA expression of IL-1β and TNF-α, showed the opposite trend (p < 0.05). In conclusion, the antioxidant capacity of PCs was stronger than that of VC and VE. The addition of PCs improved the antioxidant activity and immune function of mice. Full article

Erythropoietin-producing hepatocellular receptor A2 (EphA2) has emerged as a key mediator that promotes tumor malignant progression. EphA2 overexpression and its non-canonical signaling lead to oncogenic transformation, metabolic reprogramming, resistance to treatments, and metastasis. Therefore, strategies targeting EphA2 have been evaluated in clinical trials. However, the clinical effects were not sufficient. An anti-EphA2 monoclonal antibody (mAb), Ea₂Mab-7 (mouse IgG₁, κ), demonstrated high affinity and specificity among Eph receptors. In this study, we produced recombinant class-switched Ea₂Mab-7 variants, including Ea₂Mab-7-mG_2a (mouse IgG_2a) and Ea₂Mab-7-hG₁ (human IgG₁). Both Ea₂Mab-7-mG_2a and Ea₂Mab-7-hG₁ recognized human triple-negative breast cancer MDA-MB-231, pancreatic cancer MIA PaCa-2, and colorectal cancer HCT-15 in flow cytometry. Furthermore, both Ea₂Mab-7-mG_2a and Ea₂Mab-7-hG₁ exerted significant antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity against these tumors. In mouse xenograft models of breast, pancreatic, and colorectal cancers, both mAbs demonstrated antitumor activity. These results indicate the potential of Ea₂Mab-7 variants for the treatment of EphA2-positive cancers. Full article

Background: Crimean-Congo hemorrhagic fever (CCHF) is a severe tick-borne viral disease with a high fatality rate, and no licensed vaccines are currently available. The nucleoprotein (NP) of the Crimean-Congo hemorrhagic fever virus (CCHFV) plays a critical role in viral replication and immune recognition, making it a promising target for vaccine development. This study aimed to design and evaluate a multiepitope recombinant DNA vaccine targeting the NP of CCHFV. Methods: Cytotoxic T lymphocyte (CTL) epitopes from the NP were predicted via immunoinformatics approaches and systematically assessed for antigenicity, allergenicity, toxicity, hydrophobicity, and global population coverage. The selected epitopes were incorporated into four DNA vaccine constructs driven by a cytomegalovirus promoter, adjuvanted with human β-defensin 3 (hBD3), and fused to the reporter protein mRuby3. The constructs were evaluated in vitro using a fluorescent reporter system designed to provide a readout of TCR signaling upon the co-culture of T lymphocytes with differentiated monocytic cells expressing antigens. In vivo immunogenicity and protective efficacy were assessed in BALB/c (exploratory pilot) and IFNAR^−/− mice, a highly susceptible model for viral infection. Cytokine responses were measured to assess immunogenicity. Results: In vitro assays showed predominantly antigen-independent T-cell activation, suggesting that nonspecific stimulation inherent to the reporter co-culture system likely obscured the detection of antigen-specific TCR signaling. In vivo analyses in BALB/c mice revealed that the constructs elicited only modest systemic cytokine profiles while CCHFV-specific IgG and IFN-γ secretion remained undetectable, indicating that antigen-specific T-cell and antibody responses were limited. In the IFNAR^−/− challenge model, several peptide groups achieved significant 2–3 log reductions in tissue viral RNA and infectious titers (p < 0.05 vs. sham). However, the observed viral modulations were insufficient to reach the protective threshold and did not translate to a survival benefit (0%). Conclusion: Despite a rational in silico foundation, the multiepitope DNA vaccine constructs demonstrated limitations in inducing potent, antigen-specific immunity across both mouse models. The lack of antigen-specific responses indicates limitations in epitope selection, construct design, and delivery strategies, requiring optimization of next-generation epitope-based vaccines. These findings highlight the complexity of translating computational epitope predictions into functional vaccines, and provide benchmark data as a framework to guide future optimizations. Full article

Lactic acid bacteria (LAB) are increasingly used as functional feed additives in aquaculture. This study evaluated the effects of dietary Lactobacillus plantarum supplementation on growth performance, hepatic biochemical status, lipid-related indices, antioxidant status, and immune-related responses in Chinese tongue sole (Cynoglossus semilaevis). A total of 1620 juveniles (initial body weight 8.11 ± 0.23 g) were randomly assigned to three dietary treatments for 10 weeks: a control diet (CON) and diets supplemented with 500 mg/kg (LAB1) or 1000 mg/kg (LAB2) LAB (three replicate tanks per treatment). Compared with CON, LAB supplementation improved growth performance, with LAB2 showing significantly higher final body weight, weight gain rate, and specific growth rate. LAB altered hepatic function-related indices and reduced hepatic lipid peroxidation, as indicated by lower malondialdehyde (MDA). Hepatic lipid-related indices were also modulated, with LAB2 showing reduced total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triacylglycerol (TG), and total bile acid (TBA), together with increased high-density lipoprotein cholesterol (HDL-C). Serum immune indices showed non-linear responses, with C3, C4, and IgM increasing in LAB1 but decreasing in LAB2, whereas lysozyme showed an overall decreasing trend. qPCR analysis showed that LAB supplementation upregulated hepatic IGF-I and TGF-β1 expression, downregulated IL-8, TNF-α, and G6PD expression, and reduced FAS expression at the higher dose. Overall, dietary L. plantarum at 500–1000 mg/kg improved growth performance and was associated with changes in hepatic lipid-related indices, oxidative status, and immune-related responses in tongue sole. These results support further evaluation of L. plantarum as a functional feed additive in this species. Full article

Background/Objectives: Pathogenic mammarenaviruses cause severe hemorrhagic and neurologic disease in humans. Machupo virus (MACV), a New World (NW) mammarenavirus, causes Bolivian hemorrhagic fever in humans, and there are no approved vaccines. Methods: Here, we describe and compare the immunogenicity of three vaccines expressing the MACV glycoprotein complex (GPC) in C57BL/6 mice: a recombinant vesicular stomatitis virus (rVSV) and two different lipid nanoparticle (LNP)-encapsulated nucleoside-modified mRNA (mRNA-LNP) vaccines. The first mRNA-LNP vaccine, designated MACV mRNA, expresses the full-length MACV GPC. The second mRNA-LNP vaccine, called MACV VLP mRNA, encodes MACV GPC with appended sequences that induce the budding of virus-like particles (VLPs) with MACV GPC on the surface. This is the first description of any mRNA-LNP vaccine for MACV and the first comparison of mRNA and rVSVs as vaccine candidates for MACV. Results: We find that two doses of either MACV mRNA or MACV VLP mRNA are required for the induction of robust humoral and cellular immune responses including total MACV GPC IgG, neutralizing antibodies, cross-reactive antibodies that bind the related Junín virus GPC, and MACV-specific T-cell responses. To further investigate vaccination strategies for MACV, we also evaluated a heterologous prime-boost regimen involving the MACV mRNA vaccine coupled with the rVSV-based MACV vaccine. We find that the highest levels of MACV GPC-specific IgG and neutralizing titers were achieved when heterologous mRNA and rVSV prime-boost regimens were employed. Conclusions: These results elucidate differences in the immune response to different vaccine platforms for MACV and can inform future vaccine development for NW arenaviruses. Full article

West Nile virus (WNV) is a mosquito-borne flavivirus of growing importance for both human and equine health in Europe. Horses are highly susceptible to neurological disease and, because they share ecological exposure with humans, they represent valuable sentinels for detecting local viral circulation within a One Health framework. However, diagnosis of WNV infection in equines is complicated by the short and low-level viraemia, which limits the sensitivity of molecular assays, and by serological cross-reactivity with related flaviviruses and the confounding effects of vaccination. In this narrative review, we summarise the current diagnostic tools for WNV in horses, including direct detection methods (RT-qPCR, virus isolation, antigen detection) and indirect serological approaches (IgM and IgG ELISA, virus neutralisation tests), and discuss their practical performance and constraints in clinical and surveillance settings. We further examine equine surveillance systems, passive clinical reporting, active serosurveys and sentinel cohorts, and their integration with vector, avian and environmental monitoring. Key challenges include methodological heterogeneity, limited access to confirmatory testing and variable cross-sector data sharing. Finally, we outline future directions, highlighting the need for harmonised laboratory protocols, innovative field-deployable diagnostics, genomic surveillance and integrated, multi-source monitoring systems to strengthen early warning capacity and improve preparedness for WNV outbreaks in equine populations. Full article

Comparative evaluations of different serological methods for the diagnosis of toxoplasmosis in pregnant women remain limited, and the performance of indirect immunofluorescence assay (IFA), modified agglutination test (MAT), and chemiluminescent microparticle immunoassay (CMIA) has not been previously assessed simultaneously in this population. This study aimed to compare the performance of these three serological methods for the detection of Toxoplasma gondii antibodies in pregnant women. A total of 469 serum samples were collected from pregnant women receiving prenatal care through the Brazilian public healthcare system. Samples were tested using IFA, MAT, and CMIA for the detection of IgG and IgM antibodies. Statistical analyses included McNemar’s χ² test, Kappa agreement, and Pearson and Spearman correlation coefficients. IFA and MAT showed higher IgG seropositivity rates (53.1% and 51.2%, respectively) compared to CMIA (46.7% and 48.6%). Agreement between CMIA and IFA was moderate for IgG (Kappa = 0.51) and very strong for IgM (Kappa = 0.89). Pearson’s correlation for IgG between CMIA and IFA was moderate (r = 0.678), while Spearman’s correlation for IgM was weak. IFA and MAT demonstrated greater sensitivity for IgG detection than CMIA, while CMIA and IFA performed similarly for IgM. Conventional methods may complement automated systems to improve diagnostic accuracy in prenatal screening. Full article

Gut ischemia/reperfusion (I/R) injury releases damage-associated molecular patterns (DAMPs), such as extracellular cold-inducible RNA-binding protein (eCIRP). Milk fat globule–epidermal growth factor VIII-derived oligopeptide 3 (MOP3) is a novel peptide enabling macrophage uptake of eCIRP via αvβ3-integrin. MOP3 reduces inflammation in gut I/R, but its mechanisms are not completely understood. We hypothesized MOP3 promotes macrophage polarization toward an anti-inflammatory, M2-like phenotype in gut I/R. We induced gut I/R in mice through 60 min of superior mesenteric artery occlusion followed by 4 h of reperfusion. Intestines were evaluated for macrophage polarization by flow cytometry and immunofluorescence histology. Peritoneal cavity macrophages were isolated from mice and treated with eCIRP, MOP3, α_vβ₃-antibody, and/or naïve IgG for 4 or 24 h. Polarity was assessed by flow cytometry, qPCR, and ELISA. Compared to the sham, the M2 proportion after gut I/R decreased by 22.7%, and the M1 proportion increased by 241%. MOP3 treatment increased the M2 proportion by 64.3%, and the M1 proportion decreased by 22.7%. In eCIRP-stimulated macrophages, MOP3 treatment increased M2-like and reduced M1-like cell-surface markers, gene expression, and cytokine levels. α_vβ₃ antibody dramatically reduced MOP3′s effects. MOP3 promotes M2 polarization through α_vβ₃ integrin-mediated clearance of eCIRP, a novel mechanism whereby MOP3 reduces gut I/R injury. Full article