Search Results (347)

Background/Objectives: Tirzepatide is a dual GIP and GLP-1 receptor agonist indicated for the treatment of type 2 diabetes and obesity. Oral delivery of tirzepatide is limited by poor gastrointestinal permeability, pH-dependent solubility, and manufacturing challenges associated with high-dose absorption enhancers. Methods: This study developed an immediate-release oral tirzepatide tablet using a Quality by Design (QbD) approach. Sodium caprate (C10) was selected as the absorption enhancer based on acid-neutralizing capacity, Caco-2 permeability enhancement, and preliminary rat pharmacokinetic screening. Quality target product profile, critical quality attributes, preliminary hazard analysis, and failure mode and effects analysis identified binder/disintegrant ratios as critical material attributes and hammer milling conditions as critical process parameters. Face-centered central composite designs and multiple-response optimization (MRO) were applied to optimize dissolution, flowability, and tablet mechanical integrity. Results: The optimized binder/disintegrant composition produced benchmark-comparable dissolution profiles against oral semaglutide tablets in pH 1.2, 4.0, and 6.8 media, with f₂ values exceeding 50 for both C10 300 mg and 500 mg formulations. The optimized process yielded tablets with low friability (0.58%) and acceptable flowability (Carr’s index, 24). In beagle dogs, the C10 300 mg formulation achieved higher systemic exposure than the C10 500 mg formulation, with a C_max of 46.49 ± 23.79 ng/mL and AUC_last of 1261.03 ± 690.44 h·ng/mL. Conclusion: These results support C10-mediated oral tirzepatide delivery and QbD-based optimization for oral peptide tablets. Full article

Gingivitis, periodontitis, and stomatitis are common oral inflammatory disease affecting a large proportion of the global population. Increasing attention has recently been given to the development of health functional materials aimed at maintaining oral health and preventing microbial-associated oral disease. This study evaluated the efficacy of the lipoteichoic acid (LTA) fraction derived from the probiotic Lactiplantibacillus plantarum K8 (pLF) in preventing oral inflammation and microbial infection using the oral epithelial cell line YD-38. The results confirmed that pLF enhances the expression of interleukin-1 receptor-associated kinase M (IRAK-M), a negative regulator of Toll-like receptor (TLR) signaling, and inhibits the expression of pro-inflammatory cytokines, including C-C motif ligand 2 (CCL2), interleukin-6 (IL-6), and interleukin-8 (IL-8), in YD-38 cells stimulated with tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). Furthermore, it was demonstrated that pLF induces IRAK-M expression in a TLR2-involved manner and inhibits nuclear factor-kappa B (NF-κB) signaling, thereby reducing the expression of pro-inflammatory cytokines. pLF also exhibits oral antimicrobial efficacy by increasing the expression of the antimicrobial peptide human β-defensin 1 (hBD1) and human β-defensin 2 (hBD2) in a TLR2-involved manner and effectively inhibiting the growth of Porphyromonas gingivalis and Staphylococcus aureus in the epithelial cell associated system. Therefore, the LTA fraction derived from L. plantarum K8 represents a promising postbiotic candidate for the regulation of oral immune and microbial responses. Full article

This study was conducted to optimize the conditions for the synergistic treatment of cottonseed protein with microorganisms and enzymes and to evaluate the nutritional value and antioxidant activity of the resulting cottonseed peptides, with the ultimate goal of improving the nutritional quality of cottonseed protein. In single-factor experiments, laccase, alkaline protease, Saccharomyces cerevisiae, and Lactobacillus acidophilus were individually applied to cottonseed protein, and the optimal ranges for additive dosage, temperature, moisture content, and treatment duration were established using free gossypol, acid-soluble protein, and pH as response indicators. A Box–Behnken response surface design was subsequently adopted to perform an integrated analysis of the three responses and to determine the optimal conditions for the combined microbial–enzymatic treatment. The nutritional value and antioxidant activity of the cottonseed peptides obtained under these conditions were then evaluated. The optimal process parameters were identified as follows: microbial and enzyme dosages each at 1% (w/w), temperature of 37 °C, 37% moisture content, and treatment time of 96 h. Under the optimized conditions, the free gossypol content of the treated cottonseed protein was reduced to 67.30 mg/kg, representing a decrease of 83.69%; the acid-soluble protein content reached 29.72%, an increase of 25.86 percentage points; the reducing sugar content was 19.49 mg/g, an increase of 13.89 mg/g; and the pH dropped by 1.59 units to 4.91. Analysis of the peptide molecular weight distribution revealed that 99.61% of the cottonseed peptides had a molecular weight below 10,000 Da, and 65.45% were below 1000 Da. The peptides also exhibited excellent antioxidant capacity. In conclusion, the microbial–enzymatic synergistic treatment significantly elevated the contents of acid-soluble protein, reducing sugars, and peptides, enhanced antioxidant capacity, and reduced both free gossypol content and pH, thereby effectively improving the nutritional quality of cottonseed protein. Full article

Foodborne diseases present a serious public health challenge, causing roughly 600 million illnesses and 420,000 deaths annually. A significant portion of this impact is felt in Asia, where traditional fermented and dry-salted seafood, such as katsuobushi, budu, and peda, are dietary staples. These products rely on diverse microbial communities that determine their final safety, flavor, texture, and shelf life. Historically, research has centered on lactic acid bacteria (LAB), yet the functional contributions of non-LAB halotolerant species, including genera like Tetragenococcus, Staphylococcus, and Bacillus, are functionally important in these high-salinity niches. This review evaluates the transition from basic taxonomic surveys to mechanistic multi-omics approaches, integrating genomics, transcriptomics, proteomics, and metabolomics to decode microbial functionality under selective environmental pressures. We discuss how genomic mining using platforms such as BAGEL4 and antiSMASH can uncover biosynthetic gene clusters and antimicrobial peptides, while CARD supports antimicrobial resistance monitoring. Transcriptomic analysis reveals microbial responses to osmotic stress, low water activity, and pH fluctuations, whereas proteomic profiling links gene expression to active enzymes, stress proteins, and functional biomarkers. Metabolomics captures the chemical outcomes of fermentation, including amino acids, volatile organic compounds, spoilage markers, and biogenic amines. By merging these high-dimensional datasets with artificial intelligence, researchers can move toward predictive modeling that distinguishes biological causation from simple correlation. This shift offers a strategy to improve the safety, consistency, and resilience of traditional high-salinity fermented seafood systems. Full article

The growing threat of antimicrobial resistance necessitates alternative strategies to conventional antibiotics. Insects represent a promising source of antimicrobial peptides (AMPs) due to their potent innate immune responses. In this study, we investigated the hemolymph peptide extracts from Hermetia illucens larvae as a bioresource of infection-induced AMPs. Larvae were challenged with Gram-negative (Escherichia coli) and Gram-positive (Micrococcus flavus) bacteria, and hemolymph-derived peptides were extracted and fractionated by RP-HPLC. Peptide fractions were screened in vitro by agar diffusion assay against a panel of pathogenic strains. While extracts from uninfected larvae were essentially inactive, fractions 9–13 from infected larvae showed broad-spectrum antibacterial activity, including against multidrug-resistant clinical isolates such as carbapenem-resistant Klebsiella pneumoniae, suggesting an inducible immune response. Fractions were further analyzed by SDS-PAGE and LC-MS/MS to identify candidate AMPs, while shotgun proteomics on unfractionated extracts enabled quantitative comparison among conditions. Proteomics raw data are available via ProteomeXchange Consortium, with the dataset identifier PXD071144. In silico prediction using ProP 1.0, APD3, and CAMP_R4 supported the antimicrobial potential of candidate peptides. Overall, our integrative approach demonstrates that bacterial infection induces a diverse and bioactive immunopeptidome in H. illucens, supporting its potential as a sustainable platform for the discovery of novel antimicrobial agents. Full article

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a major cause of morbidity and mortality worldwide, particularly due to the emergence of drug-resistant strains. Membrane-active antimicrobial peptides (AMPs) represent attractive therapeutic candidates because they target bacterial envelope integrity and disrupt essential cellular processes. We evaluated two rationally designed LL-37-derived peptides: a truncated C-terminally amidated analog (LL37-1) and a modified variant incorporating N-terminal acetylation and a single D-amino acid substitution (D-LL37). Dose–response analysis demonstrated that D-LL37 exhibited greater antimycobacterial potency, with lower inhibitory concentrations of 90% (IC₉₀) and 50% (IC₅₀) values (18.40 ± 0.39 μM and 10.11 ± 0.60 μM, respectively) compared with LL37-1 (25.44 ± 0.36 μM and 15.45 ± 1.40 μM). Fluorescence-based permeability assays revealed partial membrane disruption (36% and 44% at IC₉₀ for LL37-1 and D-LL37, respectively), which was supported by ultrastructural alterations observed by scanning electron microscopy, including bacillary shortening, rough surface formation, cell clusters, and the presence of cellular debris, all of which are consistent with membrane damage. RT-qPCR analysis demonstrated significant upregulation of the P-type ATPase genes ctpF, ctpA, and ctpH following D-LL37 exposure. Collectively, these findings indicate that optimized LL-37-derived peptides exert antitubercular activity associated with envelope perturbation and coordinated activation of ion transport-related stress responses. Full article

To improve the flavor quality and antioxidant activity of walnut gluten peptides, gluten was extracted from defatted walnut meal by alkaline solubilization and acid precipitation, hydrolyzed with alkaline protease to prepare antioxidant peptides, and further modified by the Maillard reaction. The optimal sugar source was selected by single-factor experiments, and reaction conditions were optimized by response surface methodology. Peptide conformational changes were characterized by UV, fluorescence, DSC, FTIR, and SEM, while changes in amino acid composition, flavor properties, and antioxidant activity were systematically evaluated. Fructose was identified as the optimal sugar source. The optimal reaction conditions were a peptide-to-sugar ratio of 1:1.2, 78.5 °C, initial pH 7.6, and 2 h reaction time, under which the sensory score reached 8.5 and DPPH radical scavenging activity reached 66.92%. Maillard modification markedly altered peptide conformation, as shown by increased UV absorbance, decreased intrinsic fluorescence intensity with a red shift, an increase in denaturation temperature from 80 °C to 100 °C, reduced α-helix content, increased β-sheet content, and transformation of the microstructure from a loose porous morphology to dense block-like aggregates. Free amino acid content increased initially and then decreased, whereas total essential amino acids were largely retained, indicating that the overall nutritional composition was preserved. However, further evaluation of digestibility and bioavailability is required to confirm nutritional value. These findings provide a feasible strategy for improving the flavor and functional properties of walnut gluten peptides and support their high-value utilization. Full article

Background/Objective: In lung cancer treatment, increasing the concentration of antitumor drugs at the tumor site, enhancing efficacy, and reducing systemic toxicity are significant challenges. This study aims to develop an intelligent responsive polymer micelle system (GPDD) that achieves efficient accumulation and controlled release of drugs at lung tumor sites through targeted and pH-responsive design. Methods: The GPDD system is formed by the self-assembly of GE11-PEG-hyd-DOX conjugates and co-loads free DOX. This system utilizes the targeting effect of the GE11 peptide with the epidermal growth factor receptor (EGFR) to accumulate at the tumor site, while the hydrazone bond serves as a pH-responsive linker that breaks in the acidic tumor microenvironment, triggering drug release. Experiments employed CCK-8 cytotoxicity assays and tumor-bearing nude mouse models (strain not specified) for in vitro and in vivo evaluations. Results: In vitro experiments showed that GE11-modified GPDD effectively inhibited tumor cell growth. In tumor-bearing nude mouse experiments, GPDD demonstrated more significant tumor suppression effects and lower systemic toxicity compared to free DOX and unmodified PDD. Conclusions: The GPDD nanocarrier integrates targeting and pH responsiveness, improving antitumor efficacy and reducing side effects, with translational potential. The novelty of the study lies in its dual-functional design and co-loading strategy, providing new insights for tumor-targeted delivery systems. Full article

Peptide-based supramolecular hydrogels have emerged as promising biomaterials due to inherent biocompatibility, tunable self-assembly, and structural similarity to the extracellular matrix. This work describes the design, synthesis and characterization of a library of symmetrical bolaamphiphiles based on dehydropeptides, systematically varying both the dehydroamino acid residue and the linker. Aromatic and aliphatic dicarboxylic acids with distinct rigidities were employed to elucidate their influence on molecular self-assembly, hydrogelation, and functional performance. Hydrogel formation was triggered using a pH-responsive approach, and critical aggregation and gelation concentrations were determined. Morphological analysis by transmission electron microscopy revealed dense fibrillar networks with nanometer-scale fiber diameters, while rheological studies demonstrated viscoelastic behavior, tunable mechanical strength, and, in selected systems, efficient self-healing properties. The incorporation of phenylalanyldehydrophenylalanine significantly enhanced hydrogel formation, highlighting the importance of π–π interactions and hydrophobicity. Biological evaluation using HaCaT keratinocytes confirmed low cytotoxicity across the series. A representative injectable hydrogel exhibited sustained release of the anticancer drug methotrexate, governed predominantly by Fickian diffusion. These results establish clear structure–property–function relationships and demonstrate the potential of symmetrical bolaamphiphilic dehydropeptides as versatile platforms for controlled drug delivery. 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

The appearance of antibiotic-resistant strains of Helicobacter pylori (H. pylori) is leading to a decreased eradication rate of H. pylori infection. There is an urgent need to find new agents with antimicrobial mechanisms different from those of antibiotics, with therapeutic potential to clear colonization of H. pylori in the stomach. Some antimicrobial peptides (AMPs) possess bactericidal activity by enhancing the permeability of the outer membrane and damaging the integrity of the cell membrane. Bacteria are not susceptible to drug resistance through this antimicrobial mechanism. In this study, 28 short peptides containing 12 amino acid residues were designed based on nine amino acid fragments (KRIVQRIKD) from human cathelicidin LL-37, which is stable in gastric juice, and 3 amino acids were added at the C-terminus of the peptide. These designed peptides were not digested and degraded by pepsin at low pH values. The peptides were predicted using the online tool platform. Then, the strongest antimicrobial peptide, named SAMP-12aa (KRIVQRIKDVIR), was screened from 28 short peptides. Further studies found that SAMP-12aa retained anti-H. pylori activity after incubation in simulated gastric juice. The MIC and MBC of SAMP-12aa were 8 μg/mL and 32 μg/mL, respectively. SAMP-12aa showed good bactericidal kinetics. SAMP-12aa was found to have cell selectivity, penetrating and damaging bacterial cell membranes and exhibiting almost no toxicity to human cells at a relatively high concentration (128 μg/mL). Regulatory T (Treg) cells express CD25^High with immunosuppressive activity that induces immune tolerance in response to H. pylori. Molecular docking prediction revealed that SAMP-12aa could target the active center of Foxp3. Flow cytometry analysis revealed that SAMP-12aa can inhibit Foxp3 activity and downregulate CD25 protein expression on CD4⁺ T cells, thereby reducing the development and differentiation of CD4⁺Foxp3⁺CD25^High Treg cells with immunosuppressive effects. Further research revealed that the levels of the cytokine interferon-γ (IFN-γ), which activates CD8⁺ T-cell activity, were significantly elevated, and the levels of transforming growth factor-β (TGF-β), which inhibits CD8⁺ T-cell activity, were significantly reduced. The results of this study reveal that SAMP-12aa not only possesses antibacterial activity but also has immunomodulatory effects. Full article

Hydrogels are endowed with exceptional hydrophilicity and biocompatibility by their network structure, while also exhibiting soft physical properties similar to living tissues, which renders them ideal biomaterials. Responsive hydrogels—particularly those constructed from multicomponent systems including proteins, polysaccharides, peptides, and polyphenols—have emerged as a frontier research focus owing to their tunable responsiveness and controllable functional properties. In this review, hydrogel response mechanisms were categorized according to pH, ionic strength, temperature, light, enzymes, and multi-stimuli interactions. Key preparation strategies, encompassing chemical, physical, and enzymatic crosslinking, were systematically introduced. The preparation of hydrogels from various food-grade matrices, such as polysaccharide-based, protein-based, peptide-based, and polyphenol-based systems, was also summarized, with emphasis placed on how their tailored structures govern functional performance. Furthermore, innovative applications of responsive hydrogels were highlighted, including targeted delivery of nutrients and bioactive substances (e.g., probiotics, anthocyanins, vitamins) in functional foods, smart packaging and sensing for real-time freshness monitoring of meat and fruits, food quality detection through colorimetric and photothermal sensors, and 4D food printing for personalized nutrition and dysphagia-friendly foods. Full article

Microcystins (MCs), a group of potent hepatotoxins from cyanobacterial blooms, threaten global water security due to the resistance to conventional treatment processes and multi-organ toxicity to human. This study innovatively proposed a novel sequential process combining UV irradiation with biodegradation by Sphingopyxis sp. m6 for efficient microcystin-LR (MC-LR) removal. Results revealed that sequential UV-C pretreatment followed by Sphingopyxis sp. m6 biodegradation achieved complete degradation of 1 mg/L of MC-LR within 1 h of the biological phase, drastically reducing the treatment time compared to biodegradation alone (5 h). Mechanistic investigation revealed that low-dose UV-C (50 mJ/cm²) pretreatment induced MC-LR photoisomerization consistently with previously reported Adda geometric isomers. These photoisomers, along with residual parent MC-LR, were subsequently mineralized by Sphingopyxis sp. m6. Enzymatic pathway analysis confirmed a dual-pathway degradation, where Mlr enzymes processed both the native toxin and its isomeric forms, leading to a series of linearized peptides and Adda-derived products. Critically, the process achieved efficient detoxification, as confirmed by the restoration of HepG2 cell proliferation and protein phosphatase 2A activity. Moreover, response surface methodology optimized the key parameters (31.49 °C, pH of 7.36, 0.23 mg/L) for the highest degradation efficiency. This work provides an energy- and cost-efficient strategy for MC-LR remediation and elucidates the molecular mechanism of UV-induced photoisomerization facilitating subsequent biodegradation. Full article

The valorization of poultry bone by-products into high-value bioactive ingredients aligns with the principles of a sustainable circular bioeconomy. This study established an integrated process for the production, identification, and validation of bioactive antioxidant peptides from Xuefeng black-bone chicken bones (BCB). Alcalase was selected as the optimal protease due to its superior performance in both the degree of hydrolysis and antioxidant activity under the optimized conditions. Using response surface methodology (RSM), the optimal hydrolysis conditions were determined as 50 °C, pH 10.18, and 4.2 h, resulting in a hydrolysate with a hydrolysis degree of 25.10% and ABTS radical scavenging activity of 84.36%. Upon ultrafiltration, the <3 kDa fraction demonstrated a significantly higher antioxidant capacity than the crude hydrolysate. Further purification through gel filtration chromatography yielded the F3 sub-fraction (predominantly <1 kDa peptides), which exhibited the most potent activity across all four antioxidant assays conducted (ABTS, DPPH, hydroxyl radical scavenging, and reducing power). A liquid chromatography–tandem mass spectrometry (LC-MS/MS) analysis of F3 led to the identification of 21 peptide sequences. An in silico screening based on bioactivity and toxicity predictions pinpointed three promising candidates: DYPF, WDY, and FGYK. These peptides were chemically synthesized and validated to possess significant in vitro radical scavenging activities against both DPPH and hydroxyl radicals. Molecular docking simulations revealed that all three peptides could spontaneously bind to the Keap1 protein with a high affinity (binding energy < −7.0 kcal/mol), primarily through hydrogen bonds and hydrophobic interactions, suggesting a possible molecular mechanism that may involve the Keap1-Nrf2-ARE antioxidant pathway. This computational insight provides a testable hypothesis for their bioactivity, the verification of which is contingent upon future studies demonstrating their cellular delivery and intracellular action. This work not only provides a sustainable strategy for BCB utilization but also identifies potent antioxidant peptides with potential applications in functional foods and nutraceuticals. Full article

Staphylococcus aureus (S. aureus) is the most prevalent pathogen associated with subclinical mastitis, which significantly impacts dairy farming worldwide. Fluctuations in reproductive hormones, such as bovine prolactin (bPRL) and 17β-estradiol (E2), are known to compromise the innate immune response (IIR) of the mammary gland (MG). In this study, we evaluated the effects of bPRL and E2 on the effector response of primary bovine macrophages, isolated from lactating Holstein cows, challenged with S. aureus. We demonstrated that physiological concentrations of bPRL (5 ng/mL) and E2 (50 pg/mL) induced differential changes in the expression of pro-inflammatory (TNF-α, IL-6, and IL-1β) and anti-inflammatory (IL-10) cytokines, chemokines (IL-8), antimicrobial peptides (BNBD10 and S100A7), and miRNAs (miR-451, miR-155, miR-7863, miR-146a, miR-21a, Let-7a-5p, miR-30b, and miR-23a) in S. aureus-challenged macrophages. Moreover, these hormones promoted global histone H3 acetylation and the epigenetic H3K9ac mark without affecting H3K9me2 levels. Hormonal treatment also modulated histone deacetylase (HDAC) activity. Furthermore, hormonal treatment altered macrophage chemotaxis and phagocytosis. In conclusion, bPRL and E2 modulate the effector functions of bovine macrophages during S. aureus infection. This process could be associated with the regulation of histone H3 modifications, such as H3K9ac, in IIR-related genes. Full article