Search Results (4,203)

Background: The World Health Organization has identified the growing ineffectiveness of antibiotics against resistant pathogens as a global threat to public health, linked to increased morbidity and mortality. In this context, Pseudomonas aeruginosa stands out as a multidrug-resistant, biofilm-forming pathogen whose biofilm formation increases its tolerance to antimicrobials, which has driven the development of anti-virulence strategies as a therapeutic alternative. In this regard, the present study aimed to evaluate extracts and compounds from Piper species in assays targeting the inhibition of biofilm and virulence factors in Pseudomonas aeruginosa, as well as their anti-quorum sensing activity using Chromobacterium violaceum as a biosensor model. Methods: For this purpose, quorum sensing interference was first assessed through inhibition of violacein production using C. violaceum ATCC 12472 as a biosensor model. The modulation of virulence-associated phenotypes in P. aeruginosa ATCC BAA-47 was subsequently examined through inhibition of biofilm formation by crystal violet staining and spectrophotometric quantification of elastase, protease and pyocyanin production. Results: It was found that extracts from P. aduncum, P. sucrense, P. grande, and P. cumanense inhibited biofilm formation in P. aeruginosa and showed potential activity against quorum sensing in the C. violaceum model, while P. ceanothifolium exhibited only antibiofilm activity. Furthermore, hydroquinone-type compounds and benzoic acid derivatives reduced biofilm formation and virulence factors in P. aeruginosa. Conclusions: The results obtained demonstrate antibiofilm and anti-virulence activity, as well as a possible modulation of quorum sensing in model systems, suggesting that Piper species represent a promising source of bioactive compounds. Full article

Earthworms possess a highly developed innate immune system based on the coordinated activity of coelomocytes and humoral factors present in the coelomic fluid. These immune components play a central role in host defence against pathogens, maintenance of physiological homeostasis, and adaptation to environmental stressors. Coelomocytes exhibit remarkable functional and morphological diversity, including participation in phagocytosis, encapsulation, extracellular trap formation, cytotoxic responses, wound healing, and regulation of oxidative and osmotic stress. In addition, coelomic fluid contains numerous biologically active molecules, such as lysenin, coelomic cytolytic factor 1, perforin, serine proteases, lysozyme, antimicrobial peptides, and pattern recognition receptors, which contribute to cellular and humoral immune responses. Recent studies have demonstrated that earthworm coelomocytes are highly sensitive to environmental pollutants, including heavy metals, pesticides, nanomaterials, and microplastics, highlighting their importance in ecotoxicological research and soil biomonitoring. Furthermore, antifungal, antimicrobial, anti-inflammatory, antipyretic, and cytotoxic activities associated with coelomocytes and coelomic fluid suggest promising applications in agriculture, biotechnology, and pharmaceutical research. This review summarises current knowledge regarding the classification, characteristics, immune functions, toxicological responses, and applied significance of earthworm coelomocytes and coelomic fluid, with particular emphasis on their role in environmental monitoring and potential biomedical applications. Full article

Probiotics are considered promising feed additives for enhancing fish health and production performance in aquaculture. This study evaluated the effects of dietary supplementation with Sporolactobacillus laevolacticus on growth performance, feed utilization, intestinal health, and physiological responses in juvenile coho salmon (Oncorhynchus kisutch). Fish were fed a control diet or diets supplemented with S. laevolacticus at 0.89 × 10⁷, 0.90 × 10⁹, or 0.87 × 10¹¹ CFU/g for 10 weeks. Compared with the control, S. laevolacticus supplementation significantly increased final body weight, weight gain rate, specific growth rate, and protein efficiency ratio, while decreasing the feed conversion ratio (p < 0.05). It also significantly enhanced intestinal protease and α-amylase activities, improved serum biochemical and immune-related parameters, and promoted better intestinal morphology (p < 0.05). Additionally, S. laevolacticus supplementation led to elevated expression of antioxidant-related genes, reduced expression of pro-inflammatory genes, and altered gut microbial composition, characterized by a decrease in Proteobacteria and increases in Firmicutes and Lactobacillales. Among the tested dosages, 0.90 × 10⁹ CFU/g produced the most consistent improvements in growth performance, digestive function, intestinal health, antioxidant and immune responses, and gut microbial composition, and was therefore identified as the optimal supplementation level. Collectively, dietary S. laevolacticus at 0.90 × 10⁹ CFU/g improved growth performance and intestinal health in juvenile coho salmon, highlighting its potential as a probiotic candidate for coho salmon aquaculture. Full article

Ubiquitin-specific protease 22 (USP22) regulates epigenetic gene expression by deubiquitinating histone H2B (H2Bub1) and upregulating oncogenic proteins and pathways, while antagonizing p53-mediated tumor suppression. USP22 is frequently overexpressed in cancers and associated with therapy resistance and poor prognosis yet remains largely untargeted pharmacologically. Here, using a fluorescence-based USP22 deubiquitinase assay to screen the LOPAC^®1280 library, we identified β-Lapachone, a natural ortho-naphthoquinone with strong anticancer activities, as a potent USP22 inhibitor. β-Lapachone potently inhibited USP22 enzymatic activity, with a half-maximal inhibitory concentration (IC₅₀) of ~0.75 μM, and molecular docking revealed its occupation of the catalytic pocket adjacent to the USP22 active-site triad, supporting a potential binding mode. Functionally, β-Lapachone suppressed proliferation and induced apoptosis in A549 and H1299 RAS-mutant lung adenocarcinoma (LUAD) cells, while USP22 knockout conferred marked resistance, indicating partial USP22 dependence. In patient-derived LUAD models, β-Lapachone inhibited sphere formation and reduced CD133⁺ cancer stem cell populations. Notably, it synergized with cisplatin to enhance DNA damage and apoptosis. In vivo, β-Lapachone significantly suppressed tumor growth in a syngeneic KRAS-mutant/p53-Null mouse lung cancer model and further potentiated cisplatin-induced antitumor effects. Collectively, these findings identify β-Lapachone as a potent inhibitor of USP22 and validate USP22 inhibition as a key mechanism underlying its anticancer activity in LUAD cells, both in vitro and in vivo. Full article

Protease-activated receptor 1 (PAR1), a G protein-coupled receptor, plays a central role in coordinating multiple phases of cutaneous wound healing, including hemostasis, cell proliferation, migration, and extracellular matrix remodeling. Despite its therapeutic potential, PAR1-selective positive allosteric modulators (PAMs) remain limited. Here, we characterized the wound healing efficacy of gestodene, a third-generation progestin previously identified as a selective PAM of PAR1. Gestodene exhibited no intrinsic agonist activity but selectively potentiated PAR1-activating peptide (PAR1-AP)-induced calcium signaling without affecting PAR2 or PAR4 responses. Consistently, gestodene induced a concentration-dependent leftward shift in the PAR1-AP dose–response curve. Notably, gestodene enhanced PAR1-dependent cell proliferation, migration, and ERK1/2 activation, effects abolished by PAR1 knockout or pharmacological inhibition with vorapaxar in human keratinocytes (HaCaT) and dermal fibroblasts (HDF). Gestodene also potentiated the expression of wound healing-associated genes, including matrix metalloproteinases (MMP-1, -2, -3, -10), fibronectin, and type I collagen (COL1A1). In a murine wound model, topical administration of gestodene accelerated wound closure, achieving complete re-epithelialization by Day 8 and significantly enhancing collagen deposition, effects reversed by vorapaxar. Collectively, these findings demonstrate that gestodene accelerates cutaneous wound healing through PAR1-selective positive allosteric modulation and supports its potential as a drug repositioning candidate for wound repair. Full article

Background: Pycnodysostosis is a rare autosomal recessive skeletal disorder caused by biallelic pathogenic variants in CTSK, which encodes cathepsin K, a lysosomal cysteine protease required for osteoclast-mediated degradation of bone matrix. Case Report: We describe a girl with short stature, skeletal deformities, osteosclerosis, craniofacial features, clavicular dysplasia, and radiological evidence of fractures. Clinical exome sequencing identified two heterozygous CTSK variants, c.85T > C (p.Trp29Arg) and c.679A>T (p.Ile227Phe), both currently classified as variants of uncertain significance. Segregation analysis showed that the variants were inherited in trans. Computational modeling and in silico prediction tools supported a possible deleterious effect on cathepsin K structure or function. Serum cathepsin K was higher in the patient than in two age-matched controls; this result is reported as an exploratory observation only. Increased serum cathepsin K may reflect altered expression, secretion, clearance, or accumulation of dysfunctional protein, but cannot be interpreted as proof of compensatory upregulation. Conclusions: The patient’s clinical and radiographic features, the biallelic trans configuration of the CTSK variants, their rarity in population databases, and computational predictions support p.Trp29Arg and p.Ile227Phe as strong candidate disease-associated variants. Functional studies are required to confirm their effect on cathepsin K expression, maturation, and enzymatic activity. Full article

Nutgall tree anthracnose, caused primarily by Colletotrichum species, acts as a primary bottleneck restricting the sustainable development of the Rhus chinensis industry. Developing green biocontrol strategies by screening molecular targets for novel fungicides is highly imperative. A strain designated as Serratia plymuthica WF63 was isolated from healthy R. chinensis tissues. The strain exhibited broad-spectrum antifungal activity and multiple plant growth-promoting (PGP) traits, including the production of protease, cellulase, and indole-3-acetic acid (IAA). In vivo experiments revealed that S. plymuthica strain WF63 achieved a biocontrol efficacy of over 50% against anthracnose pathogens (Colletotrichum nymphaeae and C. fioriniae) and demonstrated significant plant growth-promoting effects. Gas chromatography–mass spectrometry (GC-MS) analysis, combined with in vitro toxicity validation of pure compounds, identified hexahydro-2H-pyrido [1,2-a]pyrazin-3(4H)-one as a core antifungal component in the fermentation broth, with a half maximal effective concentration $\left(E{C}_{50}\right)$ of 133.88 mg·L⁻¹ against the target pathogen. These findings not only highlight S. plymuthica strain WF63 as a promising antifungal biological agent but also suggest that the specific nitrogen-containing heterocyclic compound may serve as a candidate scaffold for further fungicide optimization, pending comprehensive ecotoxicological evaluation. Full article

The composition of low-polarity extracts obtained by sequential extraction of the aerial parts of Rhododendron adamsii Rehd. with hexane and methyl tert-butyl ether (MTBE) was investigated using GC-MS. The hexane extract was dominated by non-polar components: squalene, n-alkanes (nonacosane, hentriacontane), sesquiterpenes (trans-nerolidol, spathulenol, β-farnesene), and β-sitosterol. The subsequent MTBE extract was enriched in more polar lipids, primarily free triterpenic acids (ursolic and oleanolic acids). A critical finding was the complete absence of diterpene grayanotoxins in all tested extracts, confirming the safety of the non-polar extraction approach. In bioactivity assays, the total hexane extract demonstrated potent inhibitory activity against the SARS-CoV-2 main protease (3CLpro) with IC₅₀ values of 0.0125–0.025 mg/mL, only one order of magnitude higher than the reference inhibitor disulfiram. Fractionation revealed that the activity was distributed among free acids, bound acids, and the unsaponifiable residue, indicating a multicomponent mechanism. Importantly, none of the samples inhibited HIV-1 protease (IC₅₀ > 0.1 mg/mL), demonstrating selectivity for the cysteine protease 3CLpro over the aspartyl protease of HIV-1. These results highlight that sequential non-polar extraction of R. adamsii provides a grayanotoxin-free lipophilic complex with selective anti-SARS-CoV-2 protease activity, paving the way for bioactivity-guided identification of individual inhibitors. Full article

Cathepsin C (CTSC), also known as dipeptidyl peptidase I, is an upstream activator of serine protease networks that may promote metastatic progression through inflammatory amplification and microenvironmental remodeling. Increasing evidence suggests that CTSC contributes to cancer progression not simply as an overexpressed lysosomal protease, but as a context-dependent regulator of metastatic traits. This review summarizes the structure, maturation, and biological functions of CTSC, with emphasis on its protease-activating capacity and its links to tumor-associated inflammation. Current evidence connecting CTSC to epithelial–mesenchymal transition, extracellular matrix remodeling, neutrophil extracellular trap formation, and immune microenvironment reprogramming is then synthesized across hepatocellular carcinoma, renal cell carcinoma, breast cancer, colorectal cancer, non-small-cell lung cancer, and glioma. Available data most strongly support a pro-metastatic role for CTSC in breast cancer and colorectal cancer, whereas evidence in several other malignancies remains predominantly preclinical and mechanistically incomplete. Importantly, CTSC is better viewed as a targetable protease network hub than as a universal pan-cancer metastatic driver. The biomarker potential and therapeutic relevance of CTSC are also evaluated, with particular attention to the opportunities and limitations of current DPP-1/CTSC inhibitors and the need for tumor-specific translational strategies. Overall, CTSC represents a promising but still incompletely validated target in oncology, and future work should prioritize tissue-specific dependency, biomarker qualification, and rational combination approaches. Full article

Cottonseed hull is an abundant and low-cost cotton processing byproduct, but its feed application is severely limited by free gossypol. This study screened a gossypol-degrading fungal strain from naturally fermented cottonseed hulls and soy sauce koji, and evaluated its detoxification and feed improvement effects via solid-state fermentation. Strain TM-2 was identified as Aspergillus oryzae. It degraded over 60% of gossypol in liquid fermentation and 69.54% in cottonseed hull solid-state fermentation. Genome annotation revealed 409 CAZyme genes and key pathways for naphthalene and aromatic compound degradation. After fermentation, crude protein and acid-soluble protein were significantly increased, while cellulose, hemicellulose, lignin, neutral detergent fiber, and acid detergent fiber were notably reduced. Antioxidant activity was also greatly enhanced. Secretomic analysis identified 92 extracellular proteins, including hemicellulases, cellulases, proteases, and peptidases that jointly promoted detoxification and quality improvement. A. oryzae TM-2 efficiently degrades gossypol and improves feed quality, showing high value in fermented feed development and agricultural byproduct utilization. Full article

Cutaneous infections and chronic inflammatory wounds remain difficult to treat because antimicrobial resistance, polymicrobial biofilms, excessive protease activity, oxidative stress, and impaired barrier repair collectively reduce the effectiveness of conventional topical therapies. Plant-derived antimicrobial peptides (AMPs) and peptide-associated bioactives offer antimicrobial, antibiofilm, immunomodulatory, and tissue reparative potential; however, their clinical translation is limited by proteolytic instability, poor stratum corneum penetration, short cutaneous residence time, formulation variability, cytotoxicity risks and limited human evidence. The key research gap is the lack of an integrated translational framework linking plant-derived peptide bioactivity with polymer engineering, advanced delivery systems, skin microenvironment biology, manufacturability, and regulatory feasibility. This review aims to critically evaluate the design principles, therapeutic mechanisms, delivery platforms, and translational barriers of plant-based peptide–polymer therapeutics for cutaneous infection and inflammation. We summarize major classes of plant-derived antimicrobial peptides, including defensins, cyclotides, thionins, hevein-like peptides, snakins, lipid transfer proteins, and knottin-type scaffolds, and examine engineering strategies such as self-assembly, aromatic N-capping, PEGylation, lipidation, dendritic architectures, and stimuli-responsive conjugation. We further discuss topical matrices, nanocarriers, liposomes, electrospun fibers, and surface-tethered biomaterials as delivery platforms for improving peptide stability, local retention, and controlled release. Finally, we identify key translational bottlenecks, including selectivity, toxicity, scalability, batch reproducibility, regulatory classification, and insufficient clinical validation. Mechanism-driven peptide optimization, quality-by-design manufacturing, standardized preclinical models, and controlled clinical trials will be essential for advancing these systems toward safe and effective dermatological therapies. Full article

The Lysobacter capsici XL1 β-lytic protease (Blp) is a bacteriolytic enzyme that hydrolyzes peptide bonds in the interpeptide bridge of the peptidoglycan of Gram-positive bacteria, including antibiotic-resistant strains of pathogenic bacteria. The Blp has been extensively characterized. The only unexplored aspect is the mechanism by which this enzyme recognizes target cells. In this work, we demonstrated for the first time that the Blp structure contained a C-terminal subdomain that can be responsible for this interaction. Molecular modeling suggested a hydrophobic nature of the interaction between the Blp and peptidoglycan. Model mutant forms of the Blp, which have fewer hydrophobic areas in the C-terminal subdomain, also had fewer sites for potential interaction with the ligand. Wet lab experiments showed that these mutant Blp forms exhibited poorer binding to peptidoglycan and living Staphylococcus aureus 209P cells, resulting in decreased bacteriolytic and proteolytic activity. Amino acid residues N136 and Y160 in the C-terminal subdomain were identified and can be important for the interaction of the enzyme with target cells. Further research into the mechanism of target cell recognition by bacterial bacteriolytic proteases will enable the use of this knowledge to expand the specificity of action of these enzymes, including as antimicrobial agents for medical applications. Full article

Background/Objectives: Protein misfolding and amyloid fibril formation underlie several degenerative diseases, including Alzheimer’s disease and Parkinson’s disease. Alpha-1 antitrypsin (A1AT), a serpin protein, is particularly prone to misfolding, with polymerization and aggregation implicated in alpha-1 antitrypsin deficiency and associated hepatic and pulmonary disorders. In this study, we examined the structural changes in A1AT induced by the fluorinated alcohol, trifluoroethanol (TFE), and assessed the inhibitory effects of two natural polyphenols, amentoflavone (AMF) and theaflavin (TF), on aggregation and fibril formation. Methods: A library of selected phytocompounds was virtually screened against the crystal structure of A1AT (PDB 3NE4) using AutoDock Vina to elucidate their binding affinity towards it. Based on binding affinities, two compounds, AMF and TF, were selected for further studies. Protein aggregation was induced with TFE, and the protective effects of AMF and TF were evaluated using protease inhibitory activity, intrinsic fluorescence, turbidity, Rayleigh scattering, ANS fluorescence, and ThT fluorescence assays. Furthermore, 100 ns molecular dynamics simulation and MM-PBSA calculations were performed to assess the stability and binding interactions of the A1AT–ligand complexes. Results: Pre-treatment of A1AT with AMF or TF significantly inhibited TFE-induced aggregation in a dose-dependent manner, with AMF being consistently more effective. ThT fluorescence analysis revealed a ~60–65% decrease in aggregate formation upon treatment with polyphenols, with IC₅₀ values estimated at ~40 µM for AMF and ~50 µM for TF, both of which are statistically significant. Molecular docking and 100 ns molecular dynamics simulation also revealed stable A1AT–polyphenol interactions, with AMF exhibiting greater binding affinity and greater attenuation of solvent-induced conformational perturbation. Conclusions: Collectively, our findings show that TFE causes A1AT misfolding via a molten globule-like intermediate, resulting in fibril formation at 30–40% TFE, and natural polyphenols AMF and TF inhibited aggregation in a concentration-dependent manner. These observations suggest the potential of AMF and TF as lead scaffolds for anti-aggregation strategies, as modulators of amyloidogenic processes. Full article

Trichoderma asperellum and T. asperelloides are two cryptic species that have potential for use as biocontrol and biofertilizer (B&B) agents. Comparison of the reference genomes of the two species revealed that each species had seven chromosomes, but Trichoderma asperellum has about 1000 more genes than T. asperelloides. The number of genes coding for chitinases, cellulases, xylanases, secreted proteases, and genes involved in soil and plant health was slightly greater in T. asperellum than in T. asperelloides. Moreover, T. asperellum had five more genes than T. asperelloides involved in the synthesis of secondary metabolites like peptaibols and siderophores. The B&B genes were distributed on all the chromosomes. No duplicate genes were found for any of the enzymes searched. The investigation also revealed that T. asperellum had 15 copies of the internal transcribed spacer (ITS) region of ribosomal DNA compared to only seven copies in T. asperelloides. Further transcriptomic, proteomic, and efficacy studies are needed to determine the impact of the missing genes in T. asperelloides on its B&B activities compared to those of T. asperellum. The search for B&B genes in T. asperelloides was hindered by the lack of annotation for the genome. Thus, comparison only involves B&B genes searched in T. asperellum and whether homologs to the genes were available or missing in T. asperelloides. A comparison between additional strains of the two species is essential to show whether the data in this study apply to all intraspecies strains of the two species. Full article

Wound healing remains a persistent health problem with no definitive solution. It is crucial to characterize the complex wound healing process and the various growth factors, cytokines, and polypeptides involved. Transforming growth factor beta1 (rhTGF-β1) stimulates different cell types, providing multifunctionality in the wound healing process. Since proteins are sensitive to proteases, drug delivery systems are needed. Developed polymeric carrier systems are as important as the active substance. The carrier systems used in our study aim to contribute to wound healing in addition to the rhTGF-β1. We hypothesized that PLGA nanoparticles embedded in PVA/Chitosan (PVA/Chi) hydrogels could enhance the therapeutic effect of rhTGF-β1. PVA/Chitosan hydrogels were prepared by the freezing/thawing method. Several characterization studies (Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), texture analysis, and cell culture) were performed to investigate the potential of the prepared formulations to enhance the therapeutic effect of rhTGF-β1. Hydrogel formulations reduced the inhibitory effect of rhTGF-β1 on keratinocytes. The H5 hydrogel exhibited a proliferative effect on fibroblast cells, which play a crucial role in wound healing, resulting in a 78.8% increase compared to the control. As the PVA content in the hydrogel formulations increased, bioadhesion and viscosity also increased. Although TGF-β1 inhibited keratinocytes, it induced migration of both NIH-3T3 and HACAT cell lines. The formulations developed exhibit the potential to improve the therapeutic efficacy of rhTGF-β1 in wound healing. A small amount of the protein can have the same therapeutic efficacy and fewer side effects because the developed polymeric carrier systems contribute to the therapeutic efficacy. Full article