Search Results (226)

MicroRNAs (miRNAs) have emerged as important biomarkers for complex neurological conditions. Modifications in synaptic morphology characterize several of these disease states, indicating a possible role of miRNA in modulating synaptic formation and plasticity. Within the third-instar larvae of Drosophila melanogaster, we uncovered a functional role for highly human-conserved miR-92 in synaptogenesis of the glutamatergic peripheral nervous system. Loss of miR-92 results in underdeveloped synaptic architecture, coinciding with significantly reduced physiological activity. We demonstrate a novel role for miR-92 glial-specific expression to support synaptic growth function and plasticity. Modifications of miR-92 within glial tissue result in aberrant glial barrier properties, including an increased uptake of external dyes. Within the glia, miR-92 regulates a V-ATPase subunit (Vha55), impairing the glial cells from forming appropriate insulating layers around the nervous system. These modifications may impact how the nervous system adapts to its environment, increasing immature ‘ghost bouton’ budding and impairing responses to changes in environmental conditions. Our work highlights the importance of glial-specific miR-92 on synaptic development, affecting glial health and function through its downstream target Vha55, and demonstrates a novel mechanism for glia in synaptogenesis and homeostatic plasticity. Full article

Polyoxovanadates are a subclass of polyoxometalates (POMs) known to interact with proteins and to present anticancer, antimicrobial, and antiviral activities. Herein, we aimed to pursue the study of the breast anticancer activity of a mixed-valence polyoxovanadate, [Cl@V^V₇V^IV₈O₃₆]⁶⁻ (V₁₅) against MCF-7 and MDA-MB-231 cancer cell lines and to analyze its Ca²⁺-ATPase inhibition potential. ⁵¹V NMR and UV-Vis/NIR studies of V₁₅ indicated its stability in HEPES and RPMI media. For the Ca²⁺-ATPase activity, V₁₅ showed an IC₅₀ value of 14.2 μM and a mixed type of inhibition. The electrostatic potential map of V₁₅ and other POMs were correlated with the enzyme activity inhibition. V₁₅ also exhibited cytotoxicity against MDA-MB-231 (IC₅₀ = 17.2 μM) and MCF-7 (IC₅₀ = 15.1 μM) breast cancer cell lines. Using V₁₅ concentrations equivalent to half and 1/4 of the IC_50, it was observed that MDA-MB-231 cell migration was reduced by 90 and 70%, after 24 h, respectively. Moreover, V₁₅ caused morphological changes from fusiform to an epithelial-like (amoeboid) shape. Finally, V₁₅ induced the increase in RIPK1, MLKL, and RIPK3 gene expression, up to 3, 10, and 15-fold, respectively, pointing out that the mechanisms of cell death in the triple-negative breast cancer cell line may occur by necroptosis. Full article

Acrobeloides nanus is a cosmopolitan, parthenogenetic soil nematode that is widely distributed across various terrestrial environments, including forests, sand dunes, and agricultural lands. In Korea, this nematode was first isolated from soil collected from a potato farm. It has been used as a biological indicator for monitoring contamination caused by divalent metals such as copper and zinc. In this study, A. nanus was isolated from the soil collected from a cucumber farm, and its identity was confirmed using both morphological and molecular markers. Spray-induced gene silencing using double-stranded RNA (dsRNA) represents a promising new strategy for pest control. Here, we tested a spraying dsRNA that would specifically suppress the target genes in A. nanus. Three genes (Pat-10, Unc-87, and vATPase-B) were targeted, and their expression levels were assessed following treatment with their corresponding dsRNAs. The dsRNAs were sprayed onto the nematode diet. As the concentration of dsRNA increased, the expression levels of the target genes were significantly reduced, leading to notable nematode mortality. However, nematicidal activity varied among the three different dsRNAs. To practically assess these dsRNAs under field conditions, the dsRNAs were applied to the soil containing the nematodes by a drenching application. Significant mortality was observed in treatments with dsRNAs targeting vATPase-B or Pat-10, but not with dsRNA targeting Unc-87. To enhance nematicidal activity in soil, the dsRNAs were formulated with chitosan. This formulation significantly improved the stability of dsRNAs under soil conditions and increased their control efficacy against A. nanus. This study suggests that the drenching technique offers an effective strategy to the control of soil-dwelling nematode pests affecting agricultural crops. Full article

Endocytic uptake and lysosomal localization are suggested to be the key mechanisms underlying the toxicity of metal oxide nanoparticles (MONPs), with dissolution in the acidic milieu driving the response. In this study, we aimed to investigate if MONPs of varying solubility are similarly sequestered intracellularly, including in lysosomes and the role of the acidic lysosomal milieu on toxicity induced by copper oxide (CuO) nanoparticles (NPs), nickel oxide (NiO) NPs, aluminum oxide (Al₂O₃) NPs, and titanium dioxide (TiO₂) NPs of varying solubility in FE1 lung epithelial cells. Mitsui-7 multi-walled carbon nanotubes (MWCNTs) served as contrasts against particles. Enhanced darkfield hyperspectral imaging (EDF-HSI) with fluorescence microscopy was used to determine their potential association with lysosomes. The v-ATPase inhibitor Bafilomycin A1 (BaFA1) was used to assess the role of lysosomal acidification on toxicity. The results showed co-localization of all MONPs with lysosomes, with insoluble TiO₂ NPs showing the greatest co-localization. However, only acute toxicity induced by soluble CuO NPs was affected by the presence of BaFA1, showing a 14% improvement in relative survival. In addition, all MONPs were found to be associated with large actin aggregates; however, treatment with insoluble TiO₂ NPs, but not soluble CuO NPs, impaired the organization of F-actin and α-tubulin. These results indicate that MONPs are sequestered similarly intracellularly; however, the nature or magnitude of their toxicity is not similarly impacted by it. Future studies involving a broader variety of NPs are needed to fully understand the role of differential sequestration of NPs on cellular toxicity. Full article

Cancer cells can adapt to their surrounding microenvironment by upregulating glucose-regulated protein 78 kDa (GRP78) and vacuolar-type ATPase (V-ATPase) proteins to increase their proliferation and resilience to anticancer therapy. Therefore, targeting these proteins can obstruct cancer progression. A comprehensive computational study was conducted to investigate the inhibitory potential of four proton pump inhibitors (PPIs), dexlasnoprazole (DEX), esomeprazole (ESO), pantoprazole (PAN), and rabeprazole (RAB), against GRP78 and V-ATPase. Molecular docking revealed high-affinity scores for PPIs against both proteins. Moreover, molecular dynamics showed favorable root mean square deviation values for GRP78 and V-ATPase complexes, whereas root mean square fluctuations were high at the substrate-binding subdomains of GRP78 complexes and the α-helices of V-ATPase. Meanwhile, the radius of gyration and the surface-accessible surface area of the complexes were not significantly affected by ligand binding. Trajectory projections of the first two principal components showed similar motions of GRP78 structures and the fluctuating nature of V-ATPase structures, while the free-energy landscape revealed the thermodynamically favored GRP78-RAB and V-ATPase-DEX conformations. Furthermore, the binding free energy was −16.59 and −18.97 kcal/mol for GRP78-RAB and V-ATPase-DEX, respectively, indicating their stability. According to our findings, RAB and DEX are promising candidates for GRP78 and V-ATPase inhibition experiments, respectively. Full article

The frescoed Annunziata Oratory chapel in Riofreddo (Italy), a unique testimony to the pontificate of Martin V, sheds light on the trade routes of Ninfa in the first half of the 15th century. Despite having undergone several restorations in the past (the most recent in the 2010s), the Oratory presents serious conservation issues. At first glance, there are no evident signs of biological colonization; rather, the most obvious damage is attributed to detachments and saline efflorescence. Biological colonization at several points was identified using various diagnostic field and laboratory techniques such as ATPase point analysis, field stereoscopy in visible and UV light, culture-based and molecular approaches, Raman spectroscopy, and SEM analysis, biological colonization at several points was identified. The characterization of salt efflorescence was carried out using ion chromatography analysis. The presence of bacteria, fungi and algae, which are also linked to saline efflorescence, was observed. A clear correlation between the biological colonization and salt efflorescence composition was highlighted by our results, as well as the potential sources of microorganisms and salts via the capillary rise of groundwater. This early diagnostic approach regarding the presence of lithobionts and salt efflorescence demonstrates the complex interplay between environmental factors and microbial colonization, which can lead to biodeterioration processes. Full article

The midgut of Antheraea pernyi plays a critical role in antiviral defense. However, its transcriptional complexity remains poorly understood. Here, a full-length (FL) transcriptome atlas of A. pernyi midgut was developed by integrating PacBio Iso-Seq and RNA-seq techniques. The transcriptome sequences included 1850 novel protein-coding genes, 17,736 novel alternative isoforms, 1664 novel long non-coding RNAs (lncRNAs), and 858 transcription factors (TFs). In addition, 2471 alternative splicing (AS) events and 3070 alternative polyadenylation (APA) sites were identified. Moreover, 3426 and 4796 differentially expressed genes (DEGs) and isoforms were identified after ApNPV infection, respectively, besides the differentially expressed lncRNAs (164), TFs (171), and novel isoforms of ApRelish (1) and ApSOCS2 (4). Enrichment analyses showed that KEGG pathways related to metabolism were suppressed, whereas GO terms related to DNA synthesis and replication were induced. Furthermore, the autophagy and apoptosis pathways were significantly enriched among the upregulated genes. Protein–protein interaction network (PPI) analysis revealed the coordinated downregulation of genes involved in mitochondrial ribosomes, V-type and F-type ATPases, and oxidative phosphorylation, indicating the disruption of host energy metabolism and organelle acidification. Moreover, coordinated upregulation of genes associated with cytoplasmic ribosomes was observed, suggesting that the infection by ApNPV interferes with host translational machinery. These results show that ApNPV infection reprograms energy metabolism, biosynthetic processes, and immune response in A. pernyi midgut. Our study provides a foundation for elucidating the mechanisms of A. pernyi–virus interactions, particularly how the viruses affect host defense strategies. Full article

The “milky disease” in Chinese mitten crabs (Eriocheir sinensis), caused by Metschnikowia bicuspidata, poses significant threats to aquaculture, though its pathogenic mechanisms remain poorly understood. This study employs transcriptomic sequencing to analyze gene expression changes in Metschnikowia bicuspidata under hemocyte challenge, iron overload (1 mmol/mL), and combined stress, with functional validation through Common in Fungal Extracellular Membrane (CFEMgene) overexpression strains. Key findings reveal that (1) hemocyte challenge activated base excision repair (−log₁₀[P] = 7.58) and ribosome biogenesis pathways, indicating fungal adaptation through DNA repair and enhanced protein synthesis to counter host immune attacks (e.g., ROS-mediated damage). (2) Iron overload induced glutathione metabolism and pentose phosphate pathway enrichment, demonstrating mitigation of ferroptosis through NADPH/GSH antioxidant systems and autophagy/proteasome coordination. (3) Under combined stress, ribosome biogenesis (−log₁₀[P] = 1.3) and non-homologous end-joining pathways coordinated DNA repair with stress protein synthesis, complemented by vacuolar V-ATPase-mediated iron compartmentalization. (4) CFEM genes showed significant upregulation under hemocyte stress, with overexpression strains exhibiting enhanced biofilm formation (35% increased MTT cytotoxicity) and infectivity (40% higher infection rate), confirming CFEM domains mediate pathogenesis through iron homeostasis and virulence factor production. This work elucidates how M. bicuspidata employs metabolic reprogramming, oxidative stress responses, and CFEM-mediated iron regulation to establish infection, providing critical insights for developing targeted control strategies against milky disease. Full article

Diabetes is a risk factor for periodontitis. Increasing evidence suggests that a central player in this link is the vacuolar H+-ATPase (V-ATPase), which provides a physical and functional core for regulation by the catabolic lysosomal AMP-activated protein kinase complex (L-AMPK) and the anabolic mammalian target of rapamycin complex 1 (mTORC1). These complexes detect levels of various cellular nutrients, including glucose at the lysosome, and promote cellular responses to restore homeostasis. The high-glucose conditions of diabetes foster anabolic mTORC1 signaling that increases inflammation and inflammatory bone resorption in response to periodontal infections. Here, we review the structure and composition of V-ATPase, L-AMPK, mTORC1, and other elements of the energy-sensing platform. Mechanisms by which V-ATPase passes signals to the complexes are examined and recent data are reviewed. Current anti-bone resorptive therapeutics, bisphosphonates and denosumab, enhance the risk of medicine-related osteonecrosis of the jaw (MRONJ) and are not used to treat periodontal bone loss. Accumulating data suggest that it may be possible to target inflammatory bone resorption through agents that stimulate L-AMPK, including metformin and glucagon-like peptide-1 agonists. This approach may reduce inflammatory bone resorption without major effects on overall bone remodeling or increased risk of MRONJ. Full article

SNX11, a sorting nexin protein localized on the endosomal membrane, is an important protein closely related to protein sorting and endosomal trafficking. Previously, through a genome-wide CRISPR screening, we identified SNX11 as a critical protein for the entry of Dabie bandavirus. SNX11 deletion significantly inhibits the replication of Dabie bandavirus. We further discovered that the loss of SNX11 alters endosomal pH, potentially affecting the release process of Dabie bandavirus from endosomes to the cytoplasm. However, the mechanism by which SNX11 modulates endosomal pH and whether SNX11 deletion similarly inhibits other viruses remain to be elucidated. This study reveals that SNX11 can interact with the V1 subunit of the endosomal proton pump V-ATPase, affecting the expression level of this subunit on the endosomal membrane and thereby disrupting the assembly of V-ATPase. Additionally, we found that SNX11 deletion significantly inhibits the replication of dengue virus, hantavirus, and influenza virus. These findings suggest that SNX11 may be a key protein in the process of viral infection and could serve as a broad-spectrum antiviral target. Full article

Cardiac glycosides (CGs), a class of plant- and animal-derived compounds historically used to treat heart failure, have garnered renewed interest for their diverse pharmacological properties beyond Na⁺/K⁺-ATPase (NKA) inhibition. Recent studies reveal that CGs modulate key signaling pathways—such as NF-κB, PI3K/Akt, JAK/STAT, and MAPK—affecting processes central to cancer, viral infections, immune regulation, and neurodegeneration. In cancer, CGs induce multiple forms of regulated cell death, including apoptosis, ferroptosis, pyroptosis, and immunogenic cell death, while also inhibiting angiogenesis, epithelial–mesenchymal transition, and cell cycle progression. They demonstrate broad-spectrum antiviral activity by disrupting viral entry, replication, and mRNA processing in viruses such as HSV, HIV, influenza, and SARS-CoV-2. Immunologically, CGs regulate Th17 differentiation via RORγ signaling, although both inhibitory and agonistic effects have been reported. In the nervous system, CGs modulate neuroinflammation, support synaptic plasticity, and improve cognitive function in models of Alzheimer’s disease, epilepsy, and multiple sclerosis. Despite their therapeutic potential, clinical translation is hindered by narrow therapeutic indices and systemic toxicity. Advances in drug design and nanocarrier-based delivery are critical to unlocking CGs’ full potential as multi-target agents for complex diseases. This review synthesizes the current knowledge on the emerging roles of CGs and highlights strategies for their safe and effective repurposing. Full article

Mammalian renal intercalated cells are known for their role in acid secretion and maintaining acid–base balance. Herein, we discuss the theoretical reasons behind their development based on published data, focusing on the unique characteristics of renal intercalated cell biology that distinguish them from other mammalian cell types, while simultaneously attempting to explain the persistence of cells similar to intercalated cells throughout evolution. In addition, we traced these characteristics phylogenetically back to the simplest organisms. Intercalated cells have several functions and attributes. First, they contribute to kidney defense mechanisms in response to both infectious and non-infectious kidney damage. Second, intercalated cells are energized by V-ATPases in a manner similar to that of protozoa. Third, they possess T-antigens, which are commonly found in embryonic and cancer cells and which confer invasive abilities to these cells. Fourth, their plasticity enables the regeneration of other epithelial cells. These observations indicate that the origins of renal intercalated cells may be traceable back to amoeboid cells that originated from an evolutionary lineage including protists, or even to the last eukaryote common ancestor. The theoretical framework presented herein supports two predictions: first, that sponge amoebocytes possess membrane V-ATPase and are sensitive to bafilomycin, but not to ouabain; and second, that sponge amoebocytes—along with cells from diploblasts (such as Xenacoelomorpha), cnidarians, worms, fish and mollusk ionocytes, and the entire cell lineage containing V-ATPase, carbonic anhydrase, and anion exchangers (HCO₃⁻/Cl⁻)—have innate immunity, cellular dedifferentiation, and regeneration capabilities. Full article

Background: Verticillium wilt, which is a soil-borne vascular disease, causes serious economic losses worldwide. Various toxins secreted by V. dahliae are key factors that lead to wilt symptoms. Methods: The Vd-toxins CIA, indazole, and 3ICD were labeled with fluorescence groups, respectively, to observe the transport pathway. Transcriptome sequencing and qRT-PCR were employed to assess the expression patterns under Vd-toxin treatment. Results: AtALA1 and AtALA7 were up-regulated by V. dahliae and LC-toxins, and overexpression of either AtALA1 or AtALA7 increased Arabidopsis resistance against LC-toxins. Overexpression of AtALA1 improved the resistance of Arabidopsis to 4MBA, 3ICD, and indazole, while AtALA7 enhanced resistance to 4MBA, 3ICD, and CIA. AtALA7-overexpressing plants showed a stronger capability to transport CIA^FITC and 3ICD^5-FAM into vacuoles, while AtALA1-overexpressing plants accumulated indazole^5-FAM and 3ICD^5-FAM. Aggregation of AtALA1 and AtALA7 enhances the resistance of plants to V. dahliae. Conclusions: Arabidopsis P4-ATPase genes AtALA1 and AtALA7 mediated cell detoxification by transporting different Vd-toxins to vacuoles for degradation, thereby increasing resistance to Verticillium wilt. Full article

L-Glutamate (L-Glu) and its salt derivatives are widely used in the food industry as flavor enhancers. Although the consumption of these compounds is generally considered safe, some studies suggest that chronically consuming L-Glu may be associated with various disorders. In this study, Wistar pregnant rats were treated daily with 1 g/L of L-Glu in their drinking water throughout the gestational period. OPA-1, DRP-1, and mitofusin 2—key proteins involved in mitochondrial fusion and fission—were analyzed by Western blot. The results showed that L-Glu exposure significantly decreased DRP-1 levels, while OPA-1 and mitofusin 2 levels were unaffected. This was accompanied by a notable decrease in mitochondrial complexes III and V. The activities of Mg²⁺-ATPase and Na⁺/K⁺-ATPase were also analyzed in fetal cerebellar plasma membranes. Maternal L-Glu intake significantly increased Mg²⁺-ATPase activity. Regarding Na⁺/K⁺-ATPase, the data showed that L-Glu exposure did not modulate the protein level or its activity. However, a positive interaction with glutamate receptors was observed in both activities, although neither AMPA nor NMDA receptors appeared to be involved. These results suggest that chronic maternal L-Glu intake during gestation modulates Mg²⁺-ATPase activity and protein markers of mitochondrial dynamics in the fetal cerebellum, which could affect neonatal development. Full article

The Rare Cold-Inducible 2 (RCI2) gene encodes a conserved hydrophobic peptide that plays a crucial role in ion homeostasis, membrane stability, and responses to abiotic stress. In this study, six members of the MsRCI2 gene family were identified in Medicago sativa L., all of which contain highly conserved PMP₃ domains. Comparative collinearity analysis revealed syntenic relationships between M. sativa and M. truncatula, with each gene displaying distinct expression profiles under various stress conditions. Among them, MsRCI2B was significantly upregulated in response to salt stress. Alfalfa plants overexpressing MsRCI2B exhibited enhanced salt tolerance, as evidenced by increased antioxidant enzyme activities and reduced accumulation of malondialdehyde (MDA), hydrogen peroxide (H₂O₂), and superoxide anion (O₂⁻) compared to wild-type plants. Furthermore, the transgenic lines maintained better Na⁺/K⁺ homeostasis under salt stress, reflected by a lower Na⁺/K⁺ ratio and significantly elevated expression of key ion transport genes, including MsSOS1, MsAKT1, and MsNHX1. To elucidate the molecular mechanisms underlying MsRCI2B function, a yeast two-hybrid (Y2H) screen identified 151 potential interacting proteins. Gene Ontology (GO) enrichment analysis revealed that these interactors are mainly involved in antioxidant defense and ion transport. Further validation confirmed direct interactions between MsRCI2B and both calmodulin (CaM) and vacuola H⁺-ATPase (V-H⁺-ATPase), suggesting that MsRCI2B contributes to ion homeostasis through interactions with CaM and V-H⁺-ATPase, thereby promoting Na⁺/K⁺ balance and enhancing salt tolerance. These findings provide new insights into the role of MsRCI2B in salt stress responses and underscore its potential as a genetic target for enhancing salinity tolerance in forage crops. Full article