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Root-knot nematodes (Meloidogyne spp.) cause significant economic losses and have a profound impact on the agricultural sector, while commercial chemical nematicides have been found to cause pollution and are harmful to human health. The unique qualities and possible advantages of metal-based nanoparticles have gained considerable interest in plant disease control. This study synthesized, described, and evaluated CuONPs against the M. incognita nematode in terms of female biological cycle arrest, reduction of egg hatchability, and second juvenile mortality in pepper plants. It was done in both in vitro and open-field trials. Copper nanoparticles tested concurrently in graded concentrations (100, 75, 50, 25, and 12.5%) were evaluated. CuO nanoparticles were revealed to be effective nematode agents in the in vitro assay against eggs and second juvenile stages (J2s) of M. incognita, with a 100% concentration after 48 h of exposure, causing 87.9% larval mortality and 57.5% egg hatchability reduction. The open field experiment on pepper CuONPs application in direct irrigation showed 100% results with a concentration of 100%, an 84.4% decrease in egg masses/plant, a 92% decrease in J2s/100 g soil, and ultimately, an effect on the female's life cycle with an 83% decrease in number and an 89.6% reduction in galls/plant. Furthermore, pepper plants treated with CuONPs showed fresh and dry weights for the shoots and roots in open field trials. Finally, the safety effect of CuONPs on the growth of normal human cells was evidenced by its higher safety on normal human lung and skin cell lines (WI-38 and HFB4, respectively). CuONPs exhibited significantly effective growth parameters and strong nematicidal effects against nematodes, so they could be considered a safe and potential nanofertilizer and nematicide agent.

This study aimed to investigate the chemical composition and biological activities of Lavandula angustifolia essential oil (LAEO) cultivated in Ouezzane, Northwest Morocco. The chemical profile of LAEO was determined using gas chromatography-mass spectrometry (GC-MS), and its bioactivities, including antioxidant, antidiabetic, dermatoprotective, neuroprotective, and antibacterial properties, were evaluated through in vitro assays and molecular docking studies. GC-MS analysis identified 23 compounds, predominantly oxygenated monoterpenes, with linalyl acetate (31.78%) and linalool (16.58%) as major constituents. LAEO exhibited strong antioxidant activity in DPPH, ABTS, and ferric reducing power assays. It also displayed potent antibacterial effects, particularly against Gram-positive bacteria. For antidiabetic potential, LAEO demonstrated significant inhibition of α-amylase and α-glucosidase (IC₅₀: 131.94 ± 1.82 and 86.20 ± 1.23 µg/mL, respectively). Dermatoprotective effects were observed through tyrosinase inhibition (IC₅₀: 140.93 ± 3.30 µg/mL), while neuroprotective potential was evidenced by greater inhibition of acetylcholinesterase (AChE) than butyrylcholinesterase (BChE). These findings were further supported by in silico molecular docking analysis. In summary, LAEO presents a promising natural source of bioactive molecules with potential applications in food and pharmaceutical industries, addressing antioxidant, antidiabetic, and other therapeutic needs.

Pseudomonas aeruginosa is a highly adaptable opportunistic pathogen in diverse environments, causing plant, animal, and human infections. Its remarkable ability to resist antibiotics and deploy multiple virulence strategies is attributed to its large genome, horizontal gene transfer, and complex regulatory networks. In this study, we comprehensively investigated 15 structurally distinct phlorotannins against 18 major virulence-associated proteins, such as quorum-sensing proteins, adhesion proteins, exotoxins, siderophore receptors, secretion system components, proteases, motility, and biofilm formation. Molecular docking and 50-ns molecular dynamics simulations revealed that compounds such as 2-phloroeckol, 7-phloroeckol, phlorofucofuroeckol A, and phlorofucofuroeckol B formed strong and stable interactions with critical targets, type IV pilus biogenesis factor PilY1, ferripyoverdine receptor, and phenazine-1-carboxylate-methyltransferase, with binding free energies as low as - 12.24 kcal/mol. These compounds exhibited a wide range of non-covalent interactions, including hydrogen bonding and π-π stacking, with essential active site residues in target proteins. Drug-likeness and environmental safety assessments utilizing the pkCSM and VEGA (Q)SAR models revealed high oral bioavailability, low toxicity, minimal cytochrome P450 interactions, and mostly non-mutagenic profiles. This study reveals phlorotannins as prospective eco-friendly alternatives for reducing P. aeruginosa infection by addressing a broad spectrum of virulence factors with ecologically benign and biodegradable natural compounds.

Marine sponges (Porifera) from the Red Sea host diverse microbial communities that are integral to sponge health, nutrient cycling, and ecological resilience. Using high-throughput 16S rRNA amplicon sequencing, we characterized the bacterial diversity and functional potential across several Red Sea sponge species. Our findings revealed that these microbiomes are dominated by Alphaproteobacteria, Gammaproteobacteria, and Roseobacteraceae, with notable contributions from bacterial taxa involved in nitrogen fixation, organic matter degradation, and antimicrobial compound production. Functional predictions indicate that these symbionts support sponge nutrition, defense, and adaptation to the extreme Red Sea environment, including high salinity and temperature. Compared to sponge microbiomes from other marine regions, the Red Sea communities display unique taxonomic compositions and enhanced metabolic and defensive capacities. This highlights the essential ecological roles and potential biotechnological applications of these symbiotic assemblages. Our study underscores the significance of exploring sponge-associated microbiomes in understudied and extreme marine ecosystems. These results provide a foundation for future bioprospecting and work on adaptive mechanisms, emphasizing the value of Red Sea sponges and their microbiota for marine biotechnology and ecosystem resilience.

Canine B-cell lymphoma is a malignant hematologic neoplastic disease, with diffuse large B-cell lymphoma being the predominant subtype. This subtype highly expresses canine CD20 on its surface. Most clinical treatments are chemotherapeutic, with the standard treatment being the CHOP regimen. In immunotherapy, rituximab is effective for treating human B-cell lymphoma. However, its inability to bind natural canine CD20 renders it ineffective in treating canine B-cell lymphoma. The main objective of this study was to establish camel-canine chimeric antibodies and validate their in vitro biological activity. A Bactrian camel was immunized with the CD20 extracellular region peptide. The VHH fragment was amplified using nested PCR, ligated to the phage vector pMECS, and used to construct a phage antibody library with a capacity of 3.4 × 10¹⁰. After three rounds of enrichment and screening, 92 clones were selected for phage ELISA. Clones 4, 5, 8, 30, 43 and 46 exhibited high binding affinities for canine CD20. These strains were induced and purified using a prokaryotic expression system to obtain anti-canine CD20 single-domain antibodies. ELISA, Western blotting and flow cytometry analysis showed that the anti-canine CD20 single-domain antibody specifically binds to canine CD20. Cellular immunofluorescence validation revealed that the anti-canine CD20 single-domain antibody specifically binds to CD20 on the surface of Raji cells. The anti-canine CD20 single-domain antibody was subsequently ligated to the canine Fc fragment to construct a camel-canine chimeric antibody. The camel-canine chimeric antibody was expressed in CHO-S cells, and ELISA revealed that the chimeric antibody can specifically bind to the canine CD20 extracellular region polypeptide and the cell surface CD20 in Raji cells. In vitro killing assays revealed that the camel-canine chimeric antibody effectively mediated the targeting of canine peripheral blood mononuclear cells to Raji cells. These results indicate that the camel-canine chimeric antibody has strong anti-canine B-cell lymphoma activity and provide a basis for further development and clinical application of anti-canine B-cell lymphoma drugs.

Autism Spectrum Disorder (ASD) involves a multi-system interaction mechanism among genetics, immunity, and gut microbiota, yet its regulatory network remains undefined. This study conducted a meta-analysis on Genome-Wide Association Study data from four independent ASD cohorts to identify potential genetic loci. By integrating Polygenic Priority Score, brain region, and brain cell eQTL enrichment analyses, and combining summary-data-based Mendelian Randomisation (SMR) analyses of brain cis-eQTL and mQTL, bidirectional Mendelian Randomisation analyses of 473 gut microbiota, and SMR analysis of blood eQTL, SNPs such as rs2735307 and rs989134 with significant multi-dimensional associations were identified. These loci exert cross-tissue regulatory effects by participating in gut microbiota regulation, involving immune pathways such as T cell receptor signal activation and neutrophil extracellular trap formation, as well as cis-regulating neurodevelopmental genes (HMGN1 and H3C9P), or synergistically influencing epigenetic methylation modifications to regulate the expression of BRWD1 and ABT1. The cross-scale evidence chain constructed in this study provides a theoretical foundation for precision medicine research in ASD, holding promise to advance the development of innovative therapeutic strategies.

Heterologous overexpression of certain recombinant proteins in E. coli often triggers the formation of inclusion bodies (IBs). Traditionally viewed as inactive aggregates, IBs are now known to sometimes retain native-like folding and catalytic activity, termed catalytically active inclusion bodies (CatIBs). In this work, we investigated the CatIBs formed by recombinant LipAMS8, a cold-adapted lipase from an Antarctic Pseudomonas sp., expressed in E. coli iBL21(DE3)/pET32b. Overexpression yielded abundant insoluble aggregates which were gently isolated using 50 mM Tris-HCl, 50 mM NaCl, 1% Triton X-100 (pH 8.0), and subsequently solubilized in Tris-HCl (pH 8.0) without denaturants. Scanning electron microscopy revealed rod-like protein particles ranging from ~ 100 nm up to 1 μm. Biochemical characterization demonstrated that these LipAMS8 aggregates indeed function as CatIBs, exhibiting enzymatic activity with a defined optimum temperature, broad pH stability, specific metal ion responses, and remarkable tolerance to organic solvents. The LipAMS8 CatIBs retained significant residual activity (≥ 50%) across a wide pH range and various temperatures and showed only modest activity loss after prolonged storage (over 13 weeks at 4 °C and 8 weeks at 25 °C, maintaining > 50% activity). Notably, these aggregates displayed higher stability in extreme conditions (pH and organic media) compared to typical soluble enzymes. This study is the first to characterize a naturally formed lipase CatIB, highlighting that LipAMS8 CatIBs are produced in vivo without any artificial tags. The LipAMS8 CatIBs combine high catalytic activity with exceptional stability and solvent tolerance, underscoring an alternative strategy to obtain cold-active lipases in immobilized form. Such robust CatIB biocatalysts are highly sought after in industries for diverse biotransformations in challenging conditions.

The stability of lipase in organic solvents is crucial for biocatalytic processes in industrial biotechnology. Previously, we described a thermostable and solvent-tolerant Lip 42 from Geobacillus sp. Despite these features, the wild-type Lip 42 activity deteriorated and became unstable in methanol at high temperatures, limiting its effectiveness in solvent-driven catalysis. This study aims to integrate experimental data with molecular dynamics simulation, free energy landscape (FEL), and principal component analysis (PCA) to explore the structural and dynamic properties of V171S mutant lipase, in aqueous and methanol environments. Optimal conditions were determined at pH 8.0 and 70 °C, with notable thermal stability at 65 °C. Importantly, V171S exhibited solvent tolerance, maintaining over 70% relative activity in methanol, ethanol, acetone, 1-propanol, heptanol, octanol, and n-hexane. To further evaluate its performance in methanol, comparative in silico analyses were performed against the wild-type lipase. Structural analysis revealed that V171S maintained stability with only minor fluctuations compared to the native Lip 42 in methanol condition at 65 °C. Root mean square fluctuation (RMSF) analysis highlighted increased flexibility in the lid 1 region, suggesting structural adaptation to solvent exposure. The principal component analysis demonstrated that Lip 42 adopted broader structural distributions in methanol compared to the V171S variant. Free energy landscape analysis confirmed the presence of distinct and stable energy minima for V171S in methanol. Collectively, V171S mutation improves structural integrity under methanol stress, especially at high temperatures. This study contributes to the development of robust biocatalysts that function efficiently in mixed-solvent systems operating at elevated temperatures, especially in the field of biodiesel production.

Autoimmune liver disease (AILD) is a group of immune-mediated chronic liver injury diseases of unknown etiology. The pathogenesis of AILD is not clear, and it poses a serious threat to human life and health for a long time. Several case reports and observational studies have shown that the Epstein-Barr virus (EBV) is directly associated with AILD.However, the causal nature of the intrinsic association remains uncertain.In this study, we investigated the potential association between Epstein-Barr virus and AILDs using a two-sample Mendelian randomization analysis and generalized summary-data-based MR (GSMR) by integrating a genome-wide association study (GWAS) of five antibodies against EBV with the latest GWAS summary statistics for AILDs derived from the FinnGen database. Using summary data-based Mendelian randomization (SMR), we identified AILD-associated candidate genes linked to EBV-related single nucleotide polymorphisms (SNPs). Subsequent transcriptomic and single-cell RNA sequencing analyses were performed to elucidate their functional roles in disease pathogenesis and their interactions with immune cell populations. We found that Epstein-Barr nuclear antigen 1 (EBNA-1) may be a protective factor in autoimmune hepatitis (AIH), while our reverse MR analysis similarly found that autoimmune hepatitis may decrease the levels of EBNA-1 antibodies. More importantly, we found a positive correlation between ZEBRA antibodies and primary biliary cholangitis (PBC). In addition, according to post-GWAS analysis, five genes showed significant causal relationships with AIH and four genes with PBC, of which AIF1-positive B cells may play an important role in AIH, and M1 macrophages may play an important role in PBC progression.Our findings suggest a complex role for EBV in AILD pathogenesis, potentially acting as a double-edged sword. This study represents the first lines of genetic evidence linking EBV and AILDs, opening new avenues for understanding the etiology and potential therapeutic targets in these chronic liver conditions.