Abstract Zinc (Zn) is an essential trace element that contributes to skeletal development, bone remodeling, and mineral homeostasis, yet the molecular mechanisms underlying its effects on osteoblast differentiation remain incompletely defined. This study investigated the effects of Zn ions on preosteoblast growth and osteogenic differentiation and characterized Zn-responsive transcriptomic changes. Under osteogenic conditions, Zn increased alkaline phosphatase activity, mineralized matrix formation, and expression of osteogenic markers, including alkaline phosphatase (ALP), osteocalcin, procollagen I, and Runx2. RNA sequencing identified over a thousand differentially expressed genes following Zn²⁺ treatment. Functional enrichment analysis revealed modulation of bone-related pathways, including Wnt, TGF-β/BMP, Hippo, and PI3K-Akt signaling, as well as extracellular matrix (ECM)-receptor interaction, focal adhesion, and osteoclast differentiation. Transcriptomic analysis highlighted osteogenesis-associated candidate genes, including Wnt16, Lrp6, Bmp6, and Tnfrsf11b. Together, these findings associate Zn-enhanced osteogenic responses with changes in bone-related gene expression and identify candidate signaling relationships for future functional investigation.
Abstract Staphylococcus aureus and Escherichia coli remain major concerns in global public health, posing clinical challenges due to their adaptability, virulence, and biofilm-mediated persistence. The continued emergence of drug-resistant strains necessitates the development of new antimicrobial compounds. In this study, we evaluated the multifunctional potential of a hetero-ligated, thiolato-bridged dinuclear metallacyclic Mn(I)-based CO-releasing compound (S-MnC), analysing its antibacterial, antibiofilm, molecular, and target-binding efficacies against S. aureus and E. coli. The antibacterial potential of S-MnC was confirmed by a significant decrease in bacterial growth, biofilm biomass, matrix integrity, motility, and structural integrity. Our results further confirmed its anti-adhesive potential, demonstrating the depletion of intracellular free thiols and an increase in reactive oxygen species. Additionally, S-MnC displayed safe hemolysis levels. ADMET analysis indicated a good balance of drug-likeness properties. Molecular docking studies suggested potential binding sites of S-MnC to molecular targets such as bacterial DNA Gyrase B, SaeR, SaeS, and SarA of S. aureus, as well as DNA Gyrase B and SdiA of E. coli, suggesting the antivirulence mechanisms mediated by S-MnC. In this study, we report, for the first time, the antibacterial and antibiofilm activities of S-MnC against both Gram-negative, E. coli, and Gram-positive, S. aureus, and its binding affinity to multiple molecular and virulence targets.
Iron (Fe), zinc (Zn), manganese (Mn), and copper (Cu) are essential micronutrients for plant growth and development, but their low bioavailability in soil limits crop growth and yield. The ZIP (ZRT/IRT-like protein) family plays critical roles in the uptake and distribution of these divalent metal ions. Here, we performed a genome-wide identification of the ZIP family in tomato (Solanum lycopersicum) and analyzed their expression profiles under various metal deficiencies. A total of 14 SlZIP genes were identified and classified into three clades based on phylogenetic analysis. Conserved motif and gene structure analyses revealed that SlZIP genes within the same clade share similar exon/intron organization and protein motifs. The 14 SlZIP genes are unevenly distributed across eight chromosomes of tomato, and their promoters contain multiple iron- and copper-deficiency-responsive cis-elements. To further explore the potential functions of these SlZIP genes, we quantified their expression levels in tomato roots under metal-deficient conditions and found that SlIRT1, SlIRT2, and SlZIP4 were significantly induced by Fe deficiency, while SlZIP2 and SlZIP11 were specifically induced by Cu deficiency, and SlZIP13 and SlZIP14 by Mn deficiency. These results provide a comprehensive resource for the tomato ZIP family and lay a foundation for future functional studies of SlZIP genes in metal uptake and transport in tomato.
Metal ions play key roles in both physiological and pathological processes. The activity of nearly all nucleases depends on metal ions as cofactors. CRISPR-Cas system is widely used in gene editing and nucleic acid detection, and its ongoing development and frontier research have attracted significant attention. This study aims to explore and elucidate the effects of various metal ions on Cas12a-catalysed cleavage reactions including their specificity and regulatory mechanisms. Here, we found that certain metal ions significantly affected both the cleavage activity and the reaction specificity of Cas12a. Metal ions mediate enzyme-substrate binding by increasing the affinity between Cas12a and single-stranded DNA (ssDNA). In particular, unlike other metal ions, Mn2+ can achieve cleavage of ssDNA by only activating the catalytic domain without requiring the entire Cas12a. Structural alignment and site-directed mutagenesis of the active site revealed the structural basis of Cas12a catalysis and elucidated how metal ions influence its catalytic reaction in terms of specificity and regulatory mechanism. These results provide a theoretical foundation and offer innovative perspectives for the application of Cas12a-based enzyme technology.
Preservation and embedding procedures can substantially influence elemental distributions measured by X-ray fluorescence imaging (XFI), particularly for mobile endogenous elements. While effects of formalin fixation have been widely studied, comparatively little is known regarding methacrylate embedding. Here, larval zebrafish (Danio rerio) exposed to methylmercury-l-cysteinate (MeHg-l-Cys) were preserved either by JB-4 methacrylate embedding or using cryosectioning/air-drying, called herein cryopreservation, then were sectioned and analyzed using quantitative XFI. Mercury distributions were largely unaffected by preservation method, whereas endogenous elements including potassium, calcium, copper, sulfur, selenium, and zinc showed varying degrees of depletion or redistribution following methacrylate embedding, with potassium showing particularly strong depletion. Despite these changes, methacrylate embedding provided improved anatomical preservation relative to cryopreservation. These results demonstrate that methacrylate embedding is suitable for studies focused primarily on mercury localization, while cryopreservation remains highly preferable when accurate preservation of endogenous elemental distributions is required.
