Search Results (27,653)

Aging and Alzheimer’s disease (AD) are associated with profound changes in glial cell morphology and signaling. This study investigates the three-dimensional morphology of microglia and the intracellular localization of phosphorylated SMAD proteins as downstream effectors of transforming growth factor β (TGF-β) signaling in the amyloid precursor protein and presenilin-1 (APP/PS1) transgenic mouse model of Alzheimer’s disease. Using confocal microscopy and Simple Neurite Tracer software, we reconstructed and quantitatively analyzed glial cell morphology in aged wild-type and APP/PS1 mice. Immunofluorescence staining revealed altered pSMAD2 distribution in microglia, suggesting impaired canonical TGF-β signaling. Our findings indicate a disturbed glial morphology and dysfunctional TGF-β signaling cascade in the APP/PS1 model, underlining their potential role in Alzheimer’s disease pathogenesis. Full article

Triple-negative breast cancer (TNBC) is a highly aggressive subtype linked to a high rate of metastasis and low survival rates worldwide. Bacterial cyclodipeptides (CDPs) demonstrate anticancer properties by targeting multiple signaling pathways. The impact of CDPs on TNBC metastasis was evaluated both in vitro and in advanced-stage tumors in immunosuppressed female mice. CDPs significantly decreased the migratory and invasive capabilities of the MDA-MB-231 cell line, outperforming methotrexate (MTX). This effect was associated with the inhibition of Akt/mTOR/S6K phosphorylation, as well as Gab1, Vimentin, and FOXO1. Mice bearing MDA-MB-231 xenografts treated with CDPs alone or in combination with MTX showed near-complete suppression of primary tumors and metastatic sites in organs; notably, the combined treatment displayed a synergistic effect. Consequently, key proteins involved in tumor progression and metastasis, including p-Akt, p-Gab1, and FOXO1, were markedly inhibited in tumors from CDP-treated mice. Additionally, genes related to EMT, invasiveness, and immune modulation—including PTEN, SNAIL, CXCL1, BRCA1, GADD45A, and PD-L1—were dysregulated in the livers of TNBC-bearing mice; however, CDP treatment restored their expression more effectively than MTX. These findings suggest that the anti-metastatic effects of CDPs in the TNBC xenograft model involve modulation of the Akt/mTOR/S6K pathway, EMT, invasiveness, and immune modulation, highlighting their potential for further preclinical development. Full article

Eukaryotic initiation factor 2 (EIF2) signaling plays a crucial role in regulating mRNA translation and initiating eukaryotic protein synthesis. Computational molecular network pathway analysis of the canonical pathways of the coronaviral infection revealed that EIF2 signaling is inactivated when the coronavirus pathogenesis pathway is activated and vice versa. Our computational analyses indicated that the coronavirus pathogenesis pathway and EIF2 signaling had inverse activation states. Computational investigation of upstream or downstream microRNA (miRNA) revealed that EIF2 signaling directly interacted with miRNAs, including let-7, miR-1292-3p (miRNAs with the seed CGCGCCC), miR-15, miR-34, miR-378, miR-493, miR-497, miR-7, miR-8, and MIRLET7. A total of 36 nodes, including 8 molecules (ATF4, BCL2, CCND1, DDIT3, EIF2A, EIF2AK3, EIF4E, and ERK1/2), 1 complex (the ribosomal 40s subunit), and 1 function (apoptosis) in the coronavirus pathogenesis pathway, overlapped with EIF2 signaling. Alterations in EIF2 signaling may play a role in the pathogenesis of coronavirus. Full article

Neuropathic pain (NP) arises from maladaptive changes in peripheral and central nociceptive circuits, yet molecular alterations spanning the entire pain neuraxis remain poorly understood. Neuroinflammation is increasingly recognized as a central mechanism in NP chronification, yet the region-specific molecular events linking immune activation to affective pain processing remain inadequately defined. In this study, we employed high-resolution LC-HRMS-based quantitative proteomics to investigate chronic constriction injury (CCI)-induced molecular alterations in the sciatic nerve (SN), spinal cord (SC), and orbitofrontal cortex (OFC) of male Wistar rats, a region critical for affective and cognitive pain modulation. Behavioral assessments confirmed the development of NP phenotypes and motor deficits. Proteomic profiling revealed exclusive and differentially expressed proteins enriched in neuroinflammatory pathways across all regions. S100 proteins (S100A8 and S100B) were significantly elevated in SN, SC, and OFC, as confirmed by immunofluorescence. Their up-regulation coincided with increased astrocyte (GFAP) and microglial (Iba-1) activation, highlighting a pervasive inflammatory milieu. Intriguingly, the OFC proteome demonstrated marked up-regulation of dopamine-regulating proteins and positive regulation of dopaminergic neurotransmission, suggesting involvement of reward-related analgesic circuits. Together, our findings delineate a “nociceptive neuraxis” driven by neuroimmune activation and neuromodulatory adaptations that interfaces with dopaminergic signaling to influence sensory and affective components of pain. This integrative molecular map highlights potential therapeutic targets, including glial-derived S100 proteins and dopamine modulators for the comprehensive management of NP. Full article

With the intensification of global climate change and the frequent occurrence of extreme drought events, forest production is facing severe challenges. This study imposed drought stress and exogenous abscisic acid (ABA) treatment on Larix gmelini seedlings, evaluated their physiological characteristics, and analyzed the transcriptional response mechanism using transcriptome sequencing. The results showed that drought stress induced organ-specific changes in superoxide dismutase (SOD) and peroxidase (POD) activities, malondialdehyde (MDA) accumulation, and soluble protein content. SOD activity in leaves significantly increased, while POD activity, MDA content, and soluble protein levels in roots exhibited more dynamic changes. After ABA application, SOD activity in leaves reached its peak at 24 h, which was opposite to the situation in roots and stems, where POD activity was highest at 24 h. At 48 h, MDA accumulation was most significant in roots, while the early response in leaves was minimal. At 24 h, the soluble protein increase was most significant in stems. In addition, at this time point, ABA application significantly increased the soluble protein content in all three organs. Transcriptome sequencing analysis further identified core response genes involved in the MAPK signaling pathway, plant hormone signal transduction, starch and sucrose metabolism, and flavonoid biosynthesis pathways, including SNRK2, MAPKKK17, PYL, PP2C, XRN4, TMEM, TIR1, and TGA. In summary, Larix gmelini seedlings alleviate the inhibitory effect of drought stress on growth through a synergistic mechanism, specifically by activating the antioxidant system, initiating the MAPK signaling pathway, regulating plant hormone signal transduction, and reshaping carbon metabolism pathways, thereby enhancing stress resistance. Full article

Genomic instability not only drives tumor initiation and progression but also cooperates with apoptosis resistance to promote therapeutic evasion in hepatocellular carcinoma (HCC). Activation of MDM2, a negative regulator of p53, together with XIAP overexpression, represents a critical axis underlying this resistance. Simultaneous targeting of MDM2 and XIAP by MX69, a small molecule inhibitor, may therefore offer a potent interventional strategy to suppress cell proliferation and enhance pro-apoptotic signaling in HCC in vitro models. To evaluate the effects of MX69, cell viability was assessed via CVDK-8, colony formation, and real-time cell analysis. Oxidative stress levels and DNA damage were examined using fluorescence imaging and comet assays, respectively, while mitochondrial membrane potential was monitored through JC-1 staining. Furthermore, flow cytometry was employed to quantify apoptotic cell death and cell cycle distribution, while Western blot analysis was used to characterize the expression of apoptosis-related proteins. In vitro cytotoxicity assays revealed that MX69 reduced the viability of HUH7 and Hep3B cells in a dose-dependent manner, suppressed colony formation, and exerted anti-proliferative effects in real-time proliferation assays. Cell viability and IC50 values were evaluated using CVDK-8 and RTCA assays. Furthermore, MX69 induced oxidative stress and mitochondrial dysfunction, as evidenced by elevated ROS levels and loss of mitochondrial membrane potential. This was accompanied by significant DNA damage, detected by comet assay and γ-H2AX immunofluorescence, and G0–G1 cell cycle arrest. Moreover, MX69 triggered apoptotic cell death, demonstrating potent anticancer activity. Collectively, our findings identify MDM2/XIAP dual inhibition by MX69 as a promising therapeutic approach in HCC, with potential to overcome apoptosis resistance linked to genomic instability. Full article

Seeds possess long-lived messenger RNAs (mRNAs), some of which are involved in triggering germination and supporting seed longevity. Nevertheless, comprehensive studies on longevity-associated long-lived mRNAs in sweet corn are still scarce. To address this, eight sweet corn inbred lines were subjected to artificial aging (AA) and natural aging (NA). Based on half-inhibition time (ID₅₀), two representative lines—a high-longevity (HL, T7) and a low-longevity (LL, T3) line—were selected. Physiological and biochemical assays revealed significant reductions in superoxide dismutase (SOD) and peroxidase (POD) activities, along with increased malondialdehyde (MDA) content and electrical conductivity, with more severe membrane damage in the LL line. RNA sequencing (RNA-seq) showed a strong correlation in differentially expressed genes (DEGs) between AA and NA. The combined DEGs were enriched in mitogen-activated protein kinase (MAPK) signaling and tryptophan metabolism, while five common long-lived mRNAs, including Zm00001eb157210 and Zm00001eb164610, were consistently downregulated, suggesting their potential role in regulating seed vigor. These findings highlight key molecular players in sweet corn seed longevity. Full article

Ovarian cancer remains the most lethal gynecological malignancy, largely due to its early dissemination and extensive peritoneal metastasis. The tumor microenvironment (TME), particularly tumor-associated macrophages, promotes this invasive phenotype; however, the precise molecular effectors linking immune-to-tumor signaling remain unclear. We identified FRY, a microtubule-binding protein previously uncharacterized in ovarian pathology, as a critical mediator of macrophage-driven invasion. We observed that conditioned medium from ovarian cancer-stimulated macrophages (OCM) robustly induced FRY expression in ovarian cancer cells. Clinically, elevated FRY levels correlate with advanced tumor stage and poor patient survival. Functionally, FRY knockdown significantly abrogated OCM-induced invasion without affecting cell viability, highlighting its specific role in motility. Mechanistically, FRY facilitates epithelial–mesenchymal transition (EMT) and acts as an essential downstream effector of the PI3K/AKT signaling cascade; notably, FRY was required for AKT1-driven invasive behaviors. Furthermore, we identified the transcription factor NFIX as a key regulator of FRY expression. Macrophage-derived signals upregulate NFIX, which directly regulates FRY transcription. Pharmacological inhibition of the CXCR1/2 axis with reparixin effectively blocked OCM-mediated induction of both NFIX and FRY, suggesting that chemokine signaling initiates this pro-invasive loop. Collectively, these findings suggest that FRY is a macrophage-driven mediator of invasion and underscore its potential relevance in ovarian cancer. Full article

The immune status of dogs is shaped by continuous exposure to antigenic and various environmental stimuli, which together influence the development, regulation, and effectiveness of immune responses. Stress-related immune alterations may not be evident at the systemic level but can emerge at cellular and molecular scales. Therefore, this study aimed to comprehensively characterize the hematological and immunological profiles of dogs in different environments. We evaluated lymphocyte responses under basal conditions and following CD3/CD28-mediated in vitro activation, with subsequent long-term culture. Gene expression analyses targeted markers of early T cell activation, cytotoxic effector function, cytokine signaling, and inhibitory immune regulation. The memory phenotype of T lymphocytes was evaluated after blood collection and prolonged in vitro culture. In addition, hematological and biochemical profiles were assessed, including basic parameters, cortisol, and C-reactive protein. Our results revealed that client-owned dogs exhibited lower baseline expression of activation markers, especially in comparison with the short-term stay group, indicating an early immune activation state upon entry to the shelter environment. Furthermore, T lymphocytes from short- and long-term shelter dogs exhibited marked differences in the distribution of naïve and effector-memory subsets as well as different expansion capacity. These alterations persisted during prolonged in vitro culture, indicating that stress duration and environmental antigen exposure differentially shape immune responsiveness. In summary, chronic stress modulates canine immune status in a time-dependent manner, highlighting the importance of integrated cellular and molecular approaches in assessing the impact of environmental stressors on dogs’ health and welfare. Full article

Advanced glycation end-products (AGEs) are a variety of endogenous and exogenous substances that play an important role in inflammation, allergies, and autoimmune diseases. AGEs’ pathogenicity, alongside advanced oxidation protein products (AOPPs) and other ligands, lies in their ability to bind the receptor for advanced glycation end-products (RAGE) and trigger pro-inflammatory signaling pathways and cytokine release. The literature reports numerous studies on the role of the AGE-RAGE axis in various allergic conditions, including bronchial asthma, atopic dermatitis, food allergies, and autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, or Hashimoto’s thyroiditis, where the significant role of the AGE–RAGE axis in the immunopathogenesis of both allergic and autoimmune conditions is largely discussed and demonstrated. They suggest promising opportunities for the development of new diagnostic markers and targeted therapeutic strategies. However, further large-scale studies are needed to fully understand this multifaceted pathway and translate these insights into effective clinical interventions. Full article

Alzheimer’s disease (AD) is characterized by progressive cognitive decline, with amyloid beta oligomers (AβOs) emerging as the most neurotoxic species and acting as early triggers of cellular alterations. Before the appearance of other protein aggregates, AβOs disrupt the dynamics and stability of the neuronal cytoskeleton, a structure essential for maintaining neuronal morphology, axonal transport, and synaptic plasticity. Experimental evidence demonstrates that AβOs promote microtubule disassembly, Tau hyperphosphorylation, reduced kinesin levels, impaired axonal transport, and alterations in actin dynamics through the LIMK–cofilin signaling pathway. In addition, increased levels of neurofilament light chain have been identified as an early biomarker of axonal damage. Notably, these cytoskeletal disturbances arise in the absence of extensive neuronal death, underscoring the cytoskeleton as a critical early target in AD pathogenesis. In this review, we analyze cytoskeletal alterations induced by AβOs in neurons and discuss how these changes may contribute to disrupted neuronal communication, a defining early hallmark of AD pathology. Full article

Glyoxalase I (GLYI) is the key regulatory enzyme in the glyoxalase pathway. This pathway enables plants to neutralize methylglyoxal (MG) using glutathione (GSH), a mechanism significant for their acclimation to environmental stress. While functionally significant, the specific functions of GLYI genes in Amaranthus palmeri remain unexplored. In this study, integrated bioinformatics and expression analysis was used to identify five GLYI genes in A. palmeri. The results indicate that ApGLYI proteins are hydrophilic and slightly acidic, localized to scaffolds 1, 11, 13, and 16 of the A. palmeri genome. Phylogenetic analysis grouped ApGLYIs with other plant GLYI proteins into three distinct clades, each exhibiting conserved motif patterns. Expression analyses demonstrate that ApGLYI genes participate in both early and late regulatory phases of MG detoxification and signaling, responding to diverse stimuli including high temperature, NaCl, osmotic stress, exogenous methylglyoxal, abscisic acid (ABA), and methyl jasmonate (MeJA). Conversely, glufosinate ammonium treatment appears to compromise this cellular detoxification system. These results offer the evolutionary trajectory and functional significance of the ApGLYI gene. They establish a foundation for subsequent studies toward managing A. palmeri infestation and using these genes to improve stress resilience in cultivated crops through breeding strategies. Full article

The G protein-coupled galanin receptors include three different subtypes: galanin receptor 1, 2 and 3 (GalR1, GalR2, GalR3). The neuropeptide galanin is the principal natural agonist of the galanin receptors, the so-called galaninergic system. Galanin-like peptide and spexin have also been identified as natural ligands of the galanin receptors. Galanin receptors are widely expressed in the brain; however, they can be found in other tissues, such as the skeletal muscle, the heart, and the gastrointestinal tract. The galaninergic system regulates diverse biological processes, including feeding behavior, neuroprotection, learning, memory, cardiovascular and renal function, and nociception. Its dysregulation is associated with various diseases, such as Alzheimer’s disease, diabetes mellitus, epilepsy, depression, and cancer. The stimulation of GalR1 and GalR3 leads to the Gαi/o-type G protein-mediated inhibition of cyclic AMP/protein kinase A, whereas GalR2 stimulation initiates phospholipase C activation via Gαq/11-type G proteins. A galanin-activated β-arrestin-dependent pathway has also been described for GalR2. In this review, we summarize the recent advances concerning galanin receptor signaling, including both the G protein-dependent and -independent pathways. A better understanding of the complex interplay of the signaling molecules, receptors, and various signaling pathways is crucial for the future development of specific agonists with therapeutic potential. Full article

Annurca apple extract is gaining growing attention for its beneficial properties, particularly its outstanding antioxidant activity. Using a combination of biophysical, cell, and molecular biology techniques, this study investigates the sustainable valorization of Annurca apple by-products at different ripening stages and their role in the formation of advanced glycation end-products (AGEs), as well as in protection against AGE-related cytotoxicity. AGEs are a class of compounds formed by non-enzymatic reactions between reducing sugars and proteins, lipids, or nucleic acids. They can be produced endogenously or ingested through dietary sources and tobacco smoke. AGEs accumulate in nearly all mammalian tissues and are linked to various health issues, such as diabetes and its related complications, cardiovascular disease, and neurodegenerative disorders. Our data show that Annurca apple by-products at different ripening stages differentially counteract AGEs’ formation by inhibiting protein glycation and protect against AGE-induced cytotoxicity in endothelial cells. In particular, the extracts reduce AGE-induced reactive oxygen species (ROS) production, thereby inhibiting MAPK signaling pathways and caspase-3 activation. Moreover, ripening significantly enhances the concentration of bioactive compounds and the extent of cellular protection. This study highlights new beneficial properties of Annurca apple extracts and suggests that adopting nutritional interventions may support health and potentially reduce the risk of complications associated with AGE accumulation. Full article

Sepsis is frequently accompanied by myocardial dysfunction, which significantly worsens clinical outcomes. Lipopolysaccharide (LPS), a key component of Gram-negative bacteria, induces excessive oxidative stress and apoptosis in cardiomyocytes, contributing to sepsis-associated cardiac injury. Plasma-derived extracellular vesicles (EVs) have emerged as important mediators of intercellular communication and cardiovascular protection; however, their role in LPS-induced cardiomyocyte injury remains unclear. In this study, human AC16 cardiomyocytes were exposed to LPS in the presence or absence of plasma-derived EVs. Intracellular reactive oxygen species (ROS) production and apoptosis were assessed by flow cytometry, while apoptosis-related proteins and NF-κB signaling components were analyzed by Western blotting. The involvement of NF-κB signaling was further examined using pharmacological rescue experiments. Our results demonstrate that EV treatment markedly attenuated LPS-induced ROS accumulation and cardiomyocyte apoptosis. These protective effects were associated with reduced phosphorylation of NF-κB p65 and IκBα, as well as inhibition of p65 nuclear translocation. Notably, activation of NF-κB signaling abolished the anti-apoptotic and antioxidative effects of EVs under LPS challenge. Collectively, these findings suggest that plasma-derived EVs mitigate LPS-induced oxidative stress and apoptosis in human cardiomyocytes, potentially through modulation of NF-κB signaling. This study provides molecular insights into the cardioprotective actions of EVs and supports their potential as therapeutic candidates for sepsis-associated cardiovascular dysfunction. Full article