Search Results (162)

A novel, label-free chemiluminescence sensing platform for CpG methylation was developed, leveraging the G-quadruplex (G4) structural sensitivity of G4–protein interactions to eliminate bisulfite conversion. This sensing system is based on the enhancement of luminol chemiluminescence generated from myoglobin upon binding to the G4-forming DNA. At the core of this biosensor is the G4-structure-dependent modulation of the peroxidase-like activity generating luminol chemiluminescence of myoglobin. The structural change by CpG methylation within the G4-forming sequence of the B cell lymphoma 2 (BCL2) gene promoter altered its binding to myoglobin, transducing the methylation state into a measurable signal catalyzed by myoglobin. This principle was validated in a practical assay using immobilized probes to capture the target DNA for methylation analysis. This system demonstrated the capability to distinguish methylation differences of 50% when the target DNA concentration was over 25 nM. Versatility was further confirmed using the sequence from the dopamine receptor D2 (DRD2) gene promoter, where the methylation similarly induced distinct topological and functional changes. This is the first study to directly link the epigenetic state of a G4-forming DNA sequence to a protein-mediated enzymatic output, offering a framework for simple, rapid, and highly adaptable biosensors for research and clinical applications. Full article

Central Core Disease (CCD) is a congenital myopathy predominantly caused by mutations in the gene encoding ryanodine receptor type-1 (RYR1), the intracellular Ca²⁺ release channel embedded in the skeletal muscle sarcoplasmic reticulum membrane. The I4898T mutation represents one of the most common RYR1 mutations associated with CCD. Unfortunately, there are no approved therapies for CCD or for other myopathies caused by mutations in this gene. This study aims to perform a top-down differential proteomic analysis on soleus muscle samples from wild-type mice (WT) and heterozygous knock-in mice carrying the I4895T (IT) mutation in RyR1, to investigate the pathogenic mechanisms and molecular pathways involved in this myopathy and to shed light on new potential biomarkers useful for future therapies. Proteomic analysis revealed 50 dysregulated protein species, and multivariate analysis showed that IT mice exhibit a distinct proteomic signature compared to WT mice, characterized by alterations in proteins associated with contractile and structural dysfunction, metabolism, and stress response. In particular, a significant increase in myosin fragments was observed in IT mice, likely due to muscle breakdown. In contrast, myotilin was downregulated, suggesting a weakening of the muscle cytoskeletal structure. There was a notable downregulation of proteins involved in glycolysis and the TCA cycle; conversely, there was an increase in proteins related to anaerobic glycolysis, suggesting a shift from aerobic to anaerobic glycolysis. Furthermore, proteins involved in fatty acid beta-oxidation and oxidative phosphorylation were also found to be upregulated in IT mice, indicating an attempt by the muscle to maximize energy production. Finally, we found a significant decrease in PGC1α, which could serve as potential therapy target and biomarker in CCD. Full article

Background/Objectives: Thermoregulation is essential for survival, with the hypothalamic preoptic area integrating peripheral signals to maintain core body temperature. While fever enhances immune responses, excessive hyperthermia causes cellular damage. Previous work has shown that central glucagon-like peptide-1 (GLP-1) receptor antagonism intensifies lipopolysaccharide (LPS)-induced fever, suggesting a role for GLP-1 signaling in temperature regulation. However, the direct effects of GLP-1 receptor agonists on fever remained unexplored. This study investigated the effects of liraglutide (LIRA), a GLP-1 analog used to treat diabetes and obesity, on temperature regulation and fever in rats, with a focus on sex-dependent mechanisms. Methods: Male and female Wistar rats received lipopolysaccharide (LPS, i.p.) to induce fever, followed by LIRA treatment (0.3 mg/kg, i.p.) one hour later. Body temperature was monitored for up to six hours post-LPS injection. Results: LIRA reduced body temperature in both euthermic and febrile rats of both sexes. LPS increased PGE₂ concentration in both sexes, with males showing a twofold increase compared to females. LIRA treatment reduced PGE₂ levels in LPS-challenged males (62%, p < 0.01) but not in female rats. LPS elevated interleukin (IL)-6 levels in both sexes, while LIRA treatment decreased IL-6 only in females (45%, p < 0.05). In males, LPS reduced hypothalamic serotonin (5-HT) levels, and LIRA further decreased 5-HT in saline-treated animals. In females, LIRA increased 5-HT levels (84%, p < 0.01) in LPS-challenged animals. Additionally, LIRA exhibited sex-specific effects on hypothalamic JNK phosphorylation, increasing activation in LPS-treated males and reducing it in LPS-treated females. Conclusions: LIRA demonstrates antipyretic properties through distinct, sex-specific mechanisms. In males, temperature reduction correlates with decreased hypothalamic PGE₂, whereas in females, antipyretic effects are associated with reduced IL-6, decreased JNK phosphorylation, and increased 5-HT. These findings reveal sexually dimorphic GLP-1R-mediated thermoregulatory pathways during inflammation. However, the causal relationships between these molecular changes and temperature regulation require further investigation, particularly regarding whether observed biochemical alterations represent primary mechanisms or secondary consequences of temperature modulation. Future studies should investigate the functional significance of the apparent contradiction in serotonergic responses between sexes. Full article

The concentrations of natural radioactive elements in the groundwater environment are regulated by several factors, including aquifer geology, groundwater hydrochemical properties, and changes in environmental conditions. Many studies have explored these factors, but few have systematically elucidated the mechanisms underlying the dissolution of radioactive elements from their host minerals into groundwater. This study investigated the petrological, mineralogical, and weathering properties of aquifer materials and their effects on the leaching of uranium (U) and thorium (Th) into groundwater. The time required for the U concentration to reach the drinking water standard (30 μg/L) was estimated through artificial weathering experiments performed under diverse environmental conditions. Rock core samples were obtained from three sites differing in their geology and groundwater U concentrations. Mineralogical analyses revealed that thorite, a representative radioactive mineral that contains large amounts of U and Th, was present in samples from all collection sites. Thorite minerals differed in terms of their sizes, shapes, cracks, and chemical compositions between samples from different sites, indicating that geological features, mineral alteration characteristics, and environmental conditions controlled the behavior of U and Th. These factors appear to play crucial roles in regulating the mobility and potential long-term leachability of U and Th. Artificial weathering experiments confirmed that a neutral pH with surplus bicarbonate ions favored U leaching. Under these environmental conditions, aquifer U concentrations were estimated to require 8.7–226 years to reach the drinking water standard, depending on the groundwater dissolved oxygen content. Our results provide scientific evidence that may be used for managing radioactive elements in the groundwater environment, and are likely to inform new environmental policies and regulatory standards. Full article

2,4-Dichlorophenoxyacetic acid (2,4-D) is a widely used herbicide, yet its potential to induce hepatorenal injury via oxidative stress and apoptosis raises significant health concerns. Lycium barbarum polysaccharides (LBP) possess recognized antioxidant and anti-apoptotic properties, but their protective mechanisms against 2,4-D toxicity, particularly through a multi-target network, remain inadequately explored. This study aimed to systematically investigate the mechanisms of 2,4-D-induced hepatorenal injury and the protective efficacy of LBP by integrating network toxicology, molecular docking, and experimental validation. An integrated approach was employed. Core targets and pathways were identified via network toxicology. Molecular docking predicted interactions between 2,4-D and these targets. In vivo validation was conducted on Sprague-Dawley rats treated with 2,4-D (75 mg/kg) and/or LBP (50 mg/kg) for 28 days, assessing histopathology, serum oxidative stress markers superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA) and cellular apoptosis (TUNEL staining). Network analysis identified PPARG, NFKB1, PPARA, NFE2L2, and SERPINE1 as core targets, with molecular docking confirming strong binding affinities (binding energies: −5.1 to −6.3 kcal·mol⁻¹) and KEGG enrichment implicating cAMP, Ca²⁺, and PPAR signaling pathways. Experimentally, 2,4-D exposure induced significant histopathological damage, suppressed SOD/GSH-Px activities (p < 0.001), elevated MDA levels (p < 0.001), and markedly increased renal apoptosis (p < 0.01). Crucially, LBP intervention substantially mitigated these alterations, ameliorating tissue injury, restoring antioxidant defenses, increasing SOD/GSH-Px (p < 0.01), reducing MDA (p < 0.001) and significantly decreasing renal apoptosis (p < 0.05). This study elucidates a multi-target mechanism for 2,4-D-induced hepatorenal injury centered on oxidative stress–apoptosis dysregulation and demonstrates that LBP confers significant protection likely via modulation of this network. These findings underscore the potential of LBP as a natural protective agent against pesticide-induced organ damage and highlight the utility of integrated network approaches in toxicological research. Full article

This study explores the impact of public elderly care services on social participation among rural older adults and examines the underlying mechanisms, providing empirical evidence for improving the rural elderly care system and promoting sustainable development in rural aging societies in China. Using four waves of panel data from the China Longitudinal Aging Social Survey (CLASS) (2014–2020), this research focuses on home- and community-based elderly care services. Employing a two-way fixed-effects model and an instrumental variable approach, the study finds that the accessibility of public elderly care services significantly promotes social participation among rural older adults. This result remains robust after conducting various checks, such as replacing outcome variables, altering measurement methods, and adjusting sample sizes. Heterogeneity analysis reveals that the positive effects are more pronounced among older adults with higher education, those co-residing with adult children, and those in more economically developed regions. The accessibility of public elderly care services primarily facilitates social participation by alleviating labor supply constraints, improving mental health, and strengthening the willingness to engage. The findings suggest that improving the accessibility of public elderly care services can significantly enhance social participation and recommend expanding service coverage as a core strategy to promote active aging in rural areas, with a focus on fostering localized models like rural mutual aid and neighborhood care. Additionally, addressing information asymmetry by establishing “village-level public elderly care information service stations” and creating time banks for mutual aid care at the township level could help foster a virtuous cycle of intergenerational support. Full article

Hydrothermal fluids altered Mississippian (Osagian) carbonates in the Rebecca K. Bounds (RKB) core in western Kansas, U.S.A. Carbonate mineralization is similar to that associated with Mississippian valley type (MVT) mineralization. The RKB core displays fractures, vugs, channels, and breccias filled with saddle dolomite and blocky calcite cements. Homogenization temperature indicates that dolomite (65 to 126 °C, 18.4 to 23 wt. % NaCl) and calcite (67 to 101 °C, 13.2 to 22.4 wt. % NaCl) cements were precipitated by hot, saline fluids. These data are consistent with previous studies on the southern midcontinent. Carbon and oxygen isotope values for dolomite (δ¹³C 0.15 to 2.08‰, δ¹⁸O −6.44 to −4.66‰) and calcite (δ¹³C −1.01 to 1.79‰, δ¹⁸O −9.44 to −8.69‰) indicate multiple pulses of fluids likely sourced from basins to the south and west. Strontium isotopes data (0.7088812 to 0.7094432 in dolomite and 0.7089503 to 0.7111501 in calcite) indicate fluid interaction with granitic basement or basement-derived siliciclastics. These results are consistent with mixing of upwelling Ordovician-sourced fluids and Permian evaporitic brines, transported by advective and/or vertical migration. Although sulfide minerals were not observed in this study, earlier reports in western Kansas document sphalerite linked to hydrothermal brines in underlying strata. This study highlights the potential for MVT mineralization in the Mississippian of western Kansas. Full article

Virus-like particles (VLPs) formed as a result of self-assembly of viral capsid proteins are widely used as a platform for antigen presentation in vaccine development. However, since the inclusion of a foreign peptide into the capsid protein can alter its spatial structure and interfere with VLP assembly, such insertions are usually limited to short peptides. In this study, we have demonstrated the potential of capsid protein (CP) of single-stranded RNA phage PQ465 to present long peptides using green fluorescent protein (GFP) as a model. GFP was genetically linked to either the N- or C-terminus of PQ465 CP. Hybrid proteins were expressed in Escherichia coli and Nicotiana benthamiana plants. Spherical virus-like particles (~35 nm according to transmission electron microscopy) were successfully formed by both N- and C-terminal fusions expressed in E. coli, and by plant-produced CP with GFP fused to the C-terminus. ELISA revealed that GFP in VLPs was accessible for specific antibodies suggesting that it is exposed on the surface of PQ465-GFP particles. VLPs carrying GFP were recognized by anti-CP antibodies with less efficiency than VLPs formed by empty CP, which indicates shielding of the CP core in PQ465-GFP particles. Therefore, PQ465 CP can be used as a chimeric VLP platform for the display of relatively large protein antigens, which can operate in bacterial and plant expression systems. Full article

Background. MiR-218 is a micro-RNA expressed in two isoforms (miR-218-1 and miR-218-2) in the brain and, within the mesencephalic area, it represents a specific regulator of differentiation and functional maturation of the dopamine-releasing neurons (DAn). Deletion of miR-218 isoforms within the midbrain alters the expression of synaptic mRNAs, the neuronal excitability of DAn of the substantia nigra pars compacta (SNpc), and their ability to release dopamine (DA) within the dorsal striatum. Objectives. Here we have investigated if miR-218 impacts the function of the DAn population adjacent to SNpc, the mesencephalic ventral tegmental area (VTA) innervating the nucleus accumbens (NAcc), and the medial prefrontal cortex. Methods. With the use of miR-218-1, miR-218-2, and double conditional knock-out mice (KO1, c-KO2, c-dKO), we performed electrophysiological recordings in VTA DAn to investigate firing activity, measurements of DA release in NAcc slices by constant potential amperometry (CPA), and in vivo behavioral analysis. Results. We find that KO1 VTA neurons display hyperexcitability in comparison with c-KO2, c-dKO, and wild type (WT) neurons. DA efflux in the NAcc core and shell is reduced in all single- and double-conditional KO striatal slices in comparison with controls. The KO1 mice display a tendency toward an anxiety-like trait, as revealed by the elevated plus maze test. Conclusions. Our data indicate that miR-218-1 is the isoform that mainly regulates VTA DA neuron excitability whereas both miR-218-1 and miR-218-2 impair DA release in the mesoaccumbens pathway. Full article

Background: Primary hepatocellular carcinoma (HCC) and liver metastases differ in origin, progression, and therapeutic response, yet a direct high-resolution spatial comparison of their tumor microenvironments (TMEs) within the liver has not previously been performed. Methods: We applied high-definition spatial transcriptomics to fresh-frozen specimens of one HCC and one liver metastasis (>16,000 genes per sample, >97% mapping rates) as a proof-of-principle two-specimen study, cross-validated in human proteomics and patients’ survival datasets. Transcriptional clustering revealed spatially distinct compartments, rare cell states, and pathway alterations, which were further compared against an independent systemic dataset. Results: HCC displayed an ordered lineage architecture, with transformed hepatocyte-like tumor cells broadly dispersed across the tissue and more differentiated hepatocyte-derived cells restricted to localized zones. By contrast, liver metastases showed two sharply compartmentalized domains: an invasion zone, where proliferative stem-like tumor cells occupied TAM-rich boundaries adjacent to hypoxia-adapted tumor-core cells, and a plasticity zone, which formed a heterogeneous niche of cancer–testis antigen–positive germline-like cells. Across both tumor types, we detected a conserved metabolic program of “porphyrin overdrive,” defined by reduced cytochrome P450 expression, enhanced oxidative phosphorylation gene expression, and upregulation of FLVCR1 and ALOX5, reflecting coordinated rewiring of heme and lipid metabolism. Conclusions: In this pilot study, HCC and liver metastases demonstrated fundamentally different spatial architectures, with metastases uniquely harboring a germline/neural-like plasticity hub. Despite these organizational contrasts, both tumor types converged on a shared program of metabolic rewiring, highlighting potential therapeutic targets that link local tumor niches to systemic host–tumor interactions. Full article

The Erdaoling Mining area, located in Inner Mongolia, NW China, is recognized for its considerable potential in coalbed methane (CBM) exploration and development. However, the complex structures in this region have significant influences on coal reservoir characteristics, particularly pore structure features. This study focuses on the No. 2 coal seam of the Middle Jurassic Yan’an Formation. Three structural patterns were classified based on the existing structural characteristics of the study area. Coal samples of No. 2 coal seam were collected from different structural positions, and were subjected to low-temperature CO₂ adsorption (LTCO₂A), low-temperature N₂ adsorption/desorption (LTN₂A), low-field nuclear magnetic resonance (LF-NMR), and scanning electron microscopy (SEM) experiments, so that the structural controlling effects on pore structure would be revealed. Quantitative analysis results indicate that in terms of asymmetric syncline, from the limb to the core, the total porosity and movable fluid porosity of the coal decreased by 1.47% and 0.31%, respectively, reaching their lowest values at the core. Meanwhile, the dominant pore type shifted from primarily one-end closed pores to “ink-bottle” pores, indicating increased pore complexity. In the fold-thrust structure, the micropore specific surface area, micropore volume, mesopore specific surface area, mesopore volume, and total porosity show clear correlations with variations in coal seam structure. These parameters all reach their maximum values in the fault-cut zone at the center of the syncline, measuring 268.26 m²/g, 0.082 cm³/g, 0.601 m²/g, 1.262 cm³/g, and 4.2%, respectively. Simple pore types, like gas pores and vesicular pores, were identified in the syncline limbs, while open pores, “ink-bottle” pores, and complex multiporous types were mainly developed at fault locations, indicating that faults significantly increase the complexity of coal reservoir pore types. For the broad and gentle syncline and small-scale reverse fault combination, porosity exhibits a decreasing trend from the syncline limbs toward the core. Specifically, the mesopore specific surface area and movable fluid porosity increased by 52.24% and 43.69%, respectively, though no significant effect on micropores was observed. The syncline core in this structural setting developed normal gas pore clusters and tissue pores, with no occurrence of highly complex or heterogeneous pore types, indicating that neither the broad gentle syncline nor the small-scale faulting significantly altered the pore morphology. Comparatively, the broad and gentle syncline and small-scale reverse fault combination was determined to exert the strongest modification on pore structures of coal reservoir, followed by the asymmetric syncline, while the broad syncline alone demonstrated minimal influence. Full article

Mood disorders, including major depressive disorder (MDD) and bipolar disorder (BD), are among the leading causes of disability worldwide and are frequently associated with treatment resistance, functional impairment, and high comorbidity with metabolic dysfunction. Increasing evidence implicates insulin resistance (IR) as a key pathophysiological factor linking metabolic and psychiatric illness. IR is associated with chronic low-grade inflammation, hypothalamic–pituitary–adrenal (HPA) axis dysregulation, impaired neuroplasticity, mitochondrial dysfunction, and altered reward processing mechanisms that may contribute to core depressive features such as anhedonia, cognitive slowing, and emotional dysregulation. These processes are further exacerbated by the metabolic side effects of many psychotropic medications, creating a self-perpetuating cycle that worsens both psychiatric and physical health outcomes. Glucagon-like peptide-1 receptor agonists (GLP-1 RAs), initially developed for type 2 diabetes and obesity, have emerged as promising candidates to address this metabolic–psychiatric interface. Beyond improving glycemic control and promoting weight loss, GLP-1 RAs exert central actions relevant to mood disorders, including modulation of dopaminergic reward pathways, enhancement of hippocampal neurogenesis, attenuation of neuroinflammation, and regulation of appetite and energy balance. Preclinical studies demonstrate that GLP-1 RAs reduce microglial activation, promote hippocampal neurogenesis, and normalize stress-induced behavioral changes. Early clinical trials in patients with metabolic disorders suggest improvements in depressive symptoms, quality of life, and cognitive function, with some effects independent of weight loss or glycemic outcomes. Observational evidence also indicates reduced antidepressant use and psychological distress in diabetic and obese populations receiving GLP-1 RAs. While these findings are promising, large randomized controlled trials in primary psychiatric populations are lacking. Key challenges include clarifying dose–response relationships, disentangling central from peripheral effects, and addressing safety and adherence concerns in individuals with comorbid psychiatric conditions. Future research should focus on biomarker-informed stratification, comparative trials with standard treatments, and integration of GLP-1 RAs into multimodal care frameworks. Overall, GLP-1 RAs represent a biologically plausible and clinically relevant approach to bridging metabolic and psychiatric care, with the potential to improve outcomes in patients with mood disorders who carry a high metabolic burden. Full article

Climate change threatens coastal biodiversity, necessitating proactive conservation for endangered species like the mangrove Acanthus ebracteatus. This study integrated the MaxEnt and OPGD models to simulate its potential suitable habitats under current and three future SSP scenarios (SSP126, SSP245, and SSP585). Based on the MaxEnt model, sea surface salinity (SSS_range), sea surface temperature (SST_max), soil texture (T_silt, T_sand), and annual precipitation (Bio12) were identified as the dominant factors influencing its distribution, with SSS_range emerging as the key constraint. Furthermore, interaction analysis using the OPGD model revealed significant synergistic effects, particularly between salinity and soil properties (q > 0.8), underscoring the importance of multi-factor interactions in ecological niche modeling. Under the three SSP scenarios, the suitable habitat is projected to expand northeastward, accompanied by a poleward shift in the distribution centroid, driven predominantly by warming temperatures and altered rainfall patterns. KDE analysis revealed that existing protected areas do not fully cover regions with high habitat suitability. We propose a stratified conservation strategy that enhances in situ protection in core zones, initiates assisted restoration in potential habitats, and promotes experimental outplanting in future climatically suitable areas. This study provides scientific insights for the conservation and management of Acanthus ebracteatus under global climate change. Full article

Drought stress poses a significant threat to olive cultivation in Mediterranean regions. This study investigated the resilience and functional adaptation of root-associated and rhizosphere soil microorganisms of four olive cultivars under contrasting water regimes (irrigated vs. drought) across seasons. Using a combination of amplicon-targeted metagenomics, phylogenetic analysis, and text mining of the scientific literature, we identified a conserved core microbiome and revealed that drought stress significantly alters the structure of root-associated—but not rhizosphere soil—bacterial communities. Potential functional profiling indicated that drought conditions enriched for genes involved in stress response pathways, including branched-chain amino acid transport, glutathione S-transferase activity, thioredoxin reductase, and chemotaxis. Text mining co-occurrence networks highlighted strong associations between some key bacterial genera and plant growth-promoting functions like phytohormone production and biocontrol. Furthermore, we identified Solirubrobacter, Microvirga, and Pseudonocardia as the primary contributors to these drought-resilience functions. The stability of the soil microbiome suggests functional redundancy, whereas the restructuring of the root endophytic compartment indicates active plant selection for beneficial microbes. Our findings provide a foundation for developing tailored microbial consortia (SynComs) to enhance drought tolerance in olive trees and support sustainable agriculture in water-limited environments. Full article

Background/Objectives: An important new consideration when studying autism spectrum disorder (ASD) is the bioenergetic mechanisms underlying the relatively recent rapid evolutionary expansion of the human brain, which pose fundamental risks for mitochondrial dysfunction and calcium signaling abnormalities and their potential role in ASD, as recently highlighted by insights from the BTBR mouse model of ASD. The rapid brain expansion taking place as Homo sapiens evolved, particularly in the parietal lobe, led to increased energy demands, making the brain vulnerable to such metabolic disruptions as are seen in ASD. Methods: Mitochondrial dysfunction in ASD is characterized by impaired oxidative phosphorylation, elevated lactate and alanine levels, carnitine deficiency, abnormal reactive oxygen species (ROS), and altered calcium homeostasis. These dysfunctions are primarily functional, rather than being due to mitochondrial DNA mutations. Calcium signaling plays a crucial role in neuronal ATP production, with disruptions in inositol 1,4,5-trisphosphate receptor (ITPR)-mediated endoplasmic reticulum (ER) calcium release being observed in ASD patient-derived cells. Results: This impaired signaling affects the ER–mitochondrial calcium axis, leading to mitochondrial energy deficiency, particularly in high-energy regions of the developing brain. The BTBR mouse model, with its unique Itpr3 gene mutation, exhibits core autism-like behaviors and metabolic syndromes, providing valuable insights into ASD pathophysiology. Conclusions: Various interventions have been tested in BTBR mice, as in ASD, but none have directly targeted the Itpr3 mutation or its calcium signaling pathway. This review presents current genetic, biochemical, and neurological findings in ASD and its model systems, highlighting the need for further research into metabolic resilience and calcium signaling as potential diagnostic and therapeutic targets for ASD. Full article