Search Results (3,227)

Mitochondrial dysfunction in colonic smooth muscle cells (SMCs) is closely associated with impaired gut motility in functional constipation (FC), but the underlying molecular mechanisms remain incompletely understood. The mitochondrial unfolded protein response (UPR^mt) is a critical pathway for maintaining mitochondrial proteostasis, and heat shock factor 1 (HSF1) acts as an important upstream regulator of this response. In the present study, we employed a loperamide-induced FC mouse model, combined with single-cell transcriptomic, molecular, and functional analyses to characterize the HSF1-UPR^mt pathway in colonic SMCs and to investigate its role in FC. Single-cell transcriptomic analysis of colon tissue from FC mice revealed marked downregulation of UPR^mt-associated genes in colonic SMCs. Immunofluorescence, Western blotting, and RT-qPCR analyses of colonic tissue confirmed that HSF1 expression was reduced in colonic SMCs, along with the downregulation of the UPR^mt components, including HSP60, mtHSP70, and LONP1. These molecular changes were accompanied by mitochondrial structural damage, seen by transmission electron microscopy, and by functional impairments, including reduced mitochondrial membrane potential, elevated mtROS production, decreased ATP levels, and diminished activities of respiratory chain complexes I–V. AAV9-mediated overexpression of HSF1 reactivated the UPR^mt pathway, improved mitochondrial function, and ameliorated constipation, whereas shRNA-mediated knockdown of HSF1 further suppressed UPR^mt activity and aggravated mitochondrial damage, indicating that HSF1 bidirectionally regulates this pathway. Complementary experiments in primary colonic SMCs confirmed that this regulatory mechanism operates in a cell-autonomous manner, as modulation of HSF1 expression produced corresponding changes in the UPR^mt pathway, in the expression of mitochondrial respiratory chain complex subunits (ATP5A, NDUFA9, COX1, SDHA, UQCRC1), and in ATP production, mirroring the in vivo findings. Collectively, these results demonstrate that HSF1 plays a pivotal role in maintaining mitochondrial homeostasis in colonic SMCs through regulation of the UPR^mt pathway and that HSF1 dysfunction is closely associated with slowed gut motility in FC. These findings offer a new mechanistic perspective on FC and point to the HSF1–UPR^mt axis as a potential therapeutic target. Full article

Heat shock protein 90 (HSP90) is a molecular chaperone essential for maintaining the stability of many oncogenic client proteins. Although several HSP90 inhibitors (HSP90i) have entered clinical trials, their use has been limited by toxicity and resistance, underscoring the need for improved therapeutic strategies. In this study, we assessed the therapeutic potential of a new HSP90i, SP11, in T-cell acute lymphoblastic leukemia (T-ALL) in vitro and in the DLA mouse model in vivo, using single-cell transcriptomic profiling. Single-cell RNA sequencing showed that SP11 treatment reduces key oncogenic drivers, including MYC, BCL2, and stemness-related genes, consistent with impaired leukemic survival programs. In the DLA mouse model, SP11-mediated HSP90 inhibition was associated with alterations in the tumor microenvironment, including increased immune cell representation and enrichment of cytokine- and antigen-presentation-related transcriptional pathways. Despite these antitumor effects, a distinct subpopulation of cells continued to express or re-express MYC and BCL2, suggesting the development of early adaptive resistance. Consistent with these findings, an SP11-resistant MOLT4 cell line maintained high levels of MYC and BCL2 at both the transcript and protein levels, maintained CD44 expression, and exhibited altered inflammatory cytokine signaling. Functional studies confirmed that pharmacological inhibition of BCL2 notably increased SP11 sensitivity, supporting a rational combination strategy. Collectively, our results show that SP11 may exert both tumor-intrinsic and immune-modulating effects and reveal transcriptionally defined adaptive cellular states linked to resistance. This study provides mechanistic in sights into responses to HSP90 inhibition and supports combination approaches for improving therapeutic outcomes in T-ALL. Full article

Rare earth element-related occupational and environmental health risks have received increasing attention, but the molecular mechanisms underlying neodymium oxide (Nd₂O₃)-induced pulmonary fibrosis remain unclear. This study aimed to identify potential targets, metabolites, and pathways involved in fibrosis-related lung responses after Nd₂O₃ exposure. An integrated network toxicology and metabolomics approach was combined with an in vivo mouse model of Nd₂O₃-induced pulmonary toxicity. Histopathological injury, collagen deposition, and fibrosis-related protein expression were evaluated by HE staining, Masson staining, and Western blotting, respectively. Candidate targets were identified from public databases, key pathways were analyzed through network construction, and selected genes were validated by RT-qPCR. Nd₂O₃ exposure caused evident lung injury, inflammatory cell infiltration, structural disruption, increased collagen deposition, and elevated fibrosis-related protein expression. A total of 162 overlapping targets related to Nd₂O₃ exposure and pulmonary fibrosis were identified. ESR1, PTGS2, HSP90AA1, and MMP9 emerged as key targets, while cAMP signaling, arachidonic acid metabolism, PI3K-Akt signaling, and efferocytosis-related processes were implicated. Metabolomics showed distinct separation between control and high-exposure groups, with differential metabolites mainly associated with lipid metabolism and inflammation. RT-qPCR further confirmed altered expression of key genes. These findings suggest that Nd₂O₃ may promote fibrosis-related lung responses through inflammatory signaling, lipid metabolic disturbance, and profibrotic pathway activation. Full article

Alzheimer’s disease (AD) and type 2 diabetes mellitus (T2DM) are major global health burdens that share interconnected pathological mechanisms involving impaired insulin signaling, metabolic stress, and chronic neuroinflammation. This study applied an integrative systems biology and atomistic simulation framework to investigate bioactive compounds from Caesalpinia sappan L. targeting shared molecular regulators linking AD and T2DM. Network topology analysis identified four central hub genes, STAT3, SRC, HSP90AA1, and TP53, representing key regulatory nodes involved in inflammatory signaling, kinase regulation, proteostasis, and cellular stress responses. Compound-specific interaction analysis revealed distinct target preferences among phytochemical subclasses. Protosappanin B showed strong binding toward both STAT3 and HSP90α, whereas flavonols including quercetin and rhamnetin exhibited high affinity for SRC, and the chalcone derivative sappanchalcone preferentially interacted with TP53. Atomistic molecular dynamics simulations and MM-PBSA calculations supported stable protein ligand interactions and favorable binding energetics, while density functional theory analysis indicated electronic properties consistent with sustained intermolecular interactions. Collectively, these findings suggest that structurally distinct subclasses of C. sappan phytochemicals converge on complementary regulatory hubs within the shared AD and T2DM molecular network, supporting coordinated multi-target modulation of interconnected inflammatory, kinase signaling, proteostasis, and cellular stress pathways underlying AD–T2DM comorbidity. Full article

The Hsp20 protein family, essential in heat stress responses across all organisms, is part of the heat shock protein (Hsp) superfamily, recognized for its conserved alpha-crystallin domain (ACD). Hsp20s are the smallest proteins in the superfamily and primarily assist in protein refolding during stress and developmental processes. We present an in silico characterization of the Hsp20 gene family in Chenopodium quinoa (2n = 4x = 36) using an integrative approach. Quinoa is well known for its global contributions to food production and tolerance to various abiotic stresses. We identified 69 CqHsp20 genes that exhibit a well-conserved evolutionary pattern, characterized by a balanced copy number distributed symmetrically across 19 homeologous pairs in both subgenomes (A and B), with localized expansions driven by tandem duplications on eight chromosomes. High sequence identity in contiguous gene pairs and Ka/Ks ratios consistently below 1 (0.14–0.84) mathematically demonstrate that strict purifying selection has maintained the structural and sequence integrity of these genes since the ancestral polyploidization event. The phylogenetic analysis grouped CqHsp20 into two main clusters, splitted into four sub-clusters based on peptides’ cellular localization, consistent with a characteristic gene structure and conserved motif analysis, which may reflect the evolutionary trajectory and functional specialization of the Hsp20 family in plants. The integration of transcriptomic data from published experiments enabled us to detect a cluster of putatively ubiquitously expressed CqHsp20, as well as other groups that showed differential responses across abiotic stress conditions. The pattern shows that more genes exhibit higher transcription abundance under drought and salinity than under heat, key adaptive traits underlying quinoa’s known ecological versatility. Some of these genes, which are undetectable or have low abundance under heat stress, encode organelle-targeting peptides, a phenomenon not reported in other model plant studies. Differential expression analysis revealed a highly transcribed sub-cluster where six out of seven of nuclear CqHsp20 genes were active in aerial tissue during initial heat stress, with a specific cohort of four genes (CQ025082, CQ031384, CQ041158, and CQ055373) maintaining significant upregulation ($|{log}_2\mathrm{Fold}\mathrm{Change}|\ge 1.0$, $\mathrm{padj}<0.05$) under prolonged and simultaneous shoot/root exposure. Varying expression within CqHsp20 homologous and paralogs supports the idea that gene duplication creates genomic diversity, facilitating adaptation to variable extreme environments. However, while theoretical and in silico analysis provide valuable insight into quinoa Hsp20 response, empirical data are essential to unequivocally understand how these gene expression variations affect quinoa response to abiotic stressors. Full article

The red sea urchin Loxechinus albus is a species of high commercial importance in Chilean aquaculture, whose performance is strongly influenced by environmental conditions such as temperature. The gut microbiota plays a central role in host physiology; however, its interaction with stress-induced molecular responses remains poorly understood. This study evaluated the effects of thermal stress on food consumption, gut microbial composition, oxidative status, and immune- and stress-related gene expressions in L. albus gut. Sea urchins were exposed to control (16 °C) and elevated temperature (22 °C) conditions for 7 and 14 days. Gut microbiota was characterized using 16S rRNA sequencing, while oxidative damage to DNA and proteins was quantified. Gene expression analyses targeted markers of apoptosis (casp3, casp10, bak1), cellular growth (mtor, raptor), stress response (hsp70), and immune regulation (nfκb, foxo). Thermal stress induced a marked reduction in microbial alpha diversity and promoted a shift toward opportunistic taxa. Heat-stressed individuals exhibited significantly increased oxidative DNA damage, whereas protein oxidation remained unchanged. Gene expression analyses revealed early upregulation of casp3, casp10, nfκb, foxo, and hsp70, suggesting activation of apoptotic, immune, and stress-response pathways. In contrast, bak1, mtor, and raptor showed limited or no significant modulation. These findings demonstrate that thermal stress disrupts host–microbiota homeostasis and induces oxidative and molecular responses in L. albus. This integrative response provides insight into mechanisms underlying physiological performance under thermal stress, with important implications for aquaculture sustainability. Full article

Air pollution is a major environmental and public health issue, largely driven by human activities. The present study evaluates the combined use of two bioindicators from different taxonomic groups, the moss Rhytidiadelphus squarrosus and the terrestrial snail Cornu aspersum, to assess early biological effects induced by atmospheric exposure to toxic elements. Both species, chosen for their sensitivity, simple physiology, and suitability for field transplantation, were exposed for 30 days at two sites in southern Italy with contrasting environmental conditions. Toxic element accumulation in moss biomass and snail tissues was measured using ICP-OES, while snail shell composition was analyzed using FTIR spectroscopy. Biological responses were assessed through oxidative stress biomarkers (ROS levels and catalase activity), HSP70 expression determined by Western blotting, and structural damage, including ultrastructural changes in mosses and histopathological alterations in snails. Results showed site-dependent patterns of toxic elements accumulation in both organisms, consistent with increased oxidative stress and induction of HSP70 expression. Enlargement of the albumen gland and histological alterations in digestive tubules and reproductive systems were found in snails. Mosses showed severe ultrastructural alterations. FTIR analysis revealed changes in snail shell composition consistent with metal exposure. Principal component analysis highlighted clear patterns linking contamination, oxidative stress, and structural damage, supporting the complementarity of the two bioindicators and their ability to capture distinct exposure pathways and biological effects. Full article

Heat shock proteins (HSPs) are highly conserved molecular chaperones that play a key role in maintaining protein homeostasis and cellular survival under stress conditions. Clinically relevant human pathogenic fungi include opportunistic fungi, dimorphic fungi, dermatophytes, Mucorales, and other pathogenic groups. HSPs, including Hsp90, Hsp70, Hsp60, Hsp40, and Hsp110, are essential for the correct nascent protein folding, aggregation prevention, and degradation of misfolded polypeptides. Fungal pathogens frequently encounter environmental and host-imposed stresses, including oxidative stress, temperature fluctuations, and antifungal treatments. This review synthesizes and critically analyzes current evidence on the role of HSP families in essential processes linked to fungal virulence, including morphogenetic transitions, biofilm formation, maintenance of cell wall integrity, and interactions with host immune cells. Beyond their canonical chaperone functions, HSPs act as central mediators in pathogenic processes, such as morphogenesis transitions, biofilm formation, cell wall integrity, and interactions with host immune cells. Hsp90 stabilizes key signaling proteins involved in stress responses, morphogenesis, and antifungal resistance, while Hsp60 and Hsp70 contribute to mitochondrial function, cell wall integrity, and immune modulation. Disruption of these chaperones impairs growth, reduces virulence, and increases susceptibility to antifungal agents. The rise of antifungal resistance underscores the urgent need for new therapeutic strategies. Targeting fungal HSPs has emerged as a promising approach due to their essential roles in stress tolerance and pathogenesis. Hsp90 inhibitors, including geldanamycin derivatives and other small molecules, have demonstrated the ability to impair fungal growth, reduce virulence traits, and sensitize resistant strains to conventional antifungal drugs. Combining HSP inhibitors with existing antifungal drugs represents a potential strategy to overcome resistance and improve treatment outcomes. This review summarizes the current knowledge on HSPs in pathogenic fungi, focusing on their roles in stress adaptation, virulence, host-pathogen interaction, antifungal resistance, and their potential as targets for novel antifungal therapies. Full article

Osteoarthritis (OA) and major depressive disorder (MDD) share inflammatory and oxidative stress pathways, but the role of programmed cell death (PCD) in their comorbidity remains unclear. This study used independent OA synovial and MDD peripheral blood transcriptomic datasets—not a unified comorbid discovery cohort—to identify candidate PCD-related molecular signatures commonly dysregulated in both conditions. Transcriptomic data from OA synovium and MDD brain tissues were obtained from GEO (six training [three OA synovial and three MDD peripheral-blood], seven validation, and two single-cell RNA-seq datasets). Differentially expressed genes (DEGs) were identified, and PCD-related DEGs were screened. Machine learning (LASSO, SVM-RFE, Random Forest) was used to identify hub PCD-DEGs from the OA training set. WGCNA identified MDD-associated modules for comorbidity-gene selection. Functional enrichment, immune infiltration, scRNA-seq localization, and clinical validation (qRT-PCR/WB) were performed. From the OA cohort, four hub PCD-DEGs (CDKN1A, CX3CR1, INHBB, RHOB) showed moderate diagnostic value for OA (nomogram AUC = 0.82). Eight candidate genes (VAMP8, PDK4, P2RX4, ITM2C, IL10RA, HSP90AA1, CTSO, CRIP1) were commonly dysregulated across both OA and MDD datasets. Immune infiltration revealed upregulated B memory cells, plasma cells, Tregs, and neutrophils in OA, and neutrophils in MDD. scRNA-seq localized CDKN1A/RHOB to OA synovial cells and HSP90AA1/ITM2C to MDD neurons. Enrichment analyses highlighted TNF signaling, apoptosis, and stress responses in both diseases. An independent OA–MDD clinical cohort confirmed differential expression of CDKN1A, RHOB, ITM2C, and HSP90AA1. This study identifies four PCD-related hub genes associated with OA and eight candidate comorbidity genes showing common dysregulation across OA and MDD datasets and in an independent clinical cohort. These findings generate hypotheses about shared inflammatory pathways linking OA and MDD. As these associations derive from independent disease-specific cohorts rather than a true comorbid discovery cohort, they represent candidate signatures requiring functional validation rather than established mechanisms. Full article

Epstein–Barr virus (EBV) lytic reactivation contributes to the pathogenesis of EBV-associated epithelial malignancies, including nasopharyngeal carcinoma and gastric carcinoma, highlighting the need for therapeutic strategies targeting viral reactivation. Capsaicin exhibits anticancer and antiviral activities; however, its effects on EBV lytic reactivation remain unclear. This study investigated the effects of capsaicin on EBV lytic reactivation in EBV-positive epithelial cancer models. Capsaicin significantly suppressed the expression of lytic genes, including BZLF1, BRLF1, BMRF1, and BLLF1, and reduced EBV virion production. Proteomic analysis revealed alterations in host cellular pathways associated with metabolism, chromatin organization, and cytoskeletal regulation, whereas metabolomic profiling demonstrated perturbations in nucleotide, amino acid, and polyamine metabolism processes involved in viral DNA replication and protein synthesis. Protein–protein interaction network analysis identified key host proteins, including HSP90AB1, MYH9, and ANXA2, implicated in metabolic reprogramming, cytoskeletal organization, and stress responses. Moreover, upstream regulators associated with EBV lytic activation, including p65, AP-1, HIF-1α, and SP1, were down-regulated following capsaicin treatment. Collectively, these findings demonstrate a multitarget inhibitory effect of capsaicin on EBV lytic reactivation and support its therapeutic potential against EBV-associated epithelial malignancies. Full article

Allergic rhinitis (AR) is one of the most prevalent allergic disorders worldwide. Current pharmacological treatments are often limited by suboptimal efficacy and notable adverse effects. Herbal medicines, with their multi-component and multi-target therapeutic characteristics, have attracted increasing attention. Artemisia ordosica Krasch. (AOK), a traditional Chinese/Mongolian medicine has demonstrated immunomodulatory, antioxidant, and anti-inflammatory activities. The anti-AR potential of AOK extract fractions was evaluated using in vitro mast cell degranulation inhibition assays, network pharmacology analysis, molecular docking, and molecular dynamics simulations to elucidate underlying pharmacological mechanisms. The P815 mast cell model induced by compound 48/80 was employed to assess the inhibitory activity and cytotoxicity of different extract fractions. Among the tested fractions, the ethyl acetate fraction exhibited the most potent inhibitory effect on mast cell degranulation without significant cytotoxicity. Network pharmacology analysis identified 254 potential AR-related targets of AOK, with Signal Transducer and Activator of Transcription 3(STAT3), Src protein(SRC), Tumor protein 53(TP53), AKT Serine/Threonine Kinase 1(AKT1), Heat Shock Protein 90 Alpha Family Class A Member 1(HSP90AA1), Estrogen Receptor 1(ESR1), and Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3CA) identified as key hub proteins. Gene Ontology and KEGG pathway enrichment analyses indicated that AOK primarily modulated inflammatory and oxidative stress-related processes through the lipid and atherosclerosis, hypoxia-inducible factor-1, and AGE-RAGE signaling pathways. Molecular docking and dynamics simulations demonstrated strong binding affinities and stable interactions between major active constituents, particularly hydroxygenkwanin, and key targets such as SRC. The ethyl acetate fraction of AOK extract exhibited significant mast cell degranulation inhibitory activity, likely mediated via a synergistic multi-component, multi-target mechanism involving regulation of inflammatory and immune-related signaling pathways. These findings provide a pharmacological basis for the potential application of AOK in AR treatment. Full article

During development, stem cells rapidly proliferate and differentiate to form the embryo and the placenta, requiring intensive increases in cellular protein synthesis and changes to the cell architecture. Chaperone proteins, including the small heat shock proteins (HSPs), are critical assistants to protein folding, preventing protein aggregation, and promoting autophagy. Mitogen-activated protein kinase kinase kinase 4 (MAP3K4) is a stress-activated kinase that promotes fetal and placental growth. MAP3K4 directly activates p38 and JNK in trophoblast stem (TS) cells by phosphorylating MAP2K3 and MAP2K4/7, respectively. In addition, MAP3K4 promotes activation of the Akt signaling pathway by controlling Igf1r expression. TS cells differentiate to placental trophoblasts comprising the junctional zone (JZ) and labyrinth (LAB) placental layers. In this study, we demonstrate that JZ differentiation transiently increases JNK activity, whereas LAB differentiation induces sustained p38, JNK, and Akt activation. Each of these pathways is inhibited in MAP3K4 kinase-inactive (KI) LAB^KI trophoblasts. JZ and LAB differentiation also induces HSP22 and HSP27 expression and HSP27 phosphorylation; these are also reduced in TS^KI and LAB^KI cells. JZ and LAB differentiation induces GABARAP-positive autophagosomes that are deficient in KI cells. Altogether, our findings demonstrate that MAP3K4 is critical for responses during differentiation in placental trophoblasts. Full article

Coastal ecosystems are increasingly threatened by human activities, highlighting the need for sensitive tools to assess environmental risk. An active biomonitoring approach, using the Mediterranean mussel (Mytilus galloprovincialis), was employed to evaluate anthropogenic chemical contamination in the North Ionian Sea, a still poorly studied area, by comparing mussel health status before (PrePT) and after (PostPT) the peak tourist season. Bioaccumulation of metal(loid)s was quantified in whole organisms. Oxidative stress was assessed in the gills and digestive gland through catalase (CAT), superoxide dismutase (SOD), lipid peroxidation (LPO), and oxidized carbonyl proteins (OMP). Neurotoxicity was evaluated via acetylcholinesterase (AChE) activity, while gene expression of stress-related biomarkers was analysed for metallothioneins (mt10, mt20), sod, cat, Glutathione S-transferase (gst), and Heat Shock Protein 70 (hsp70). Results suggest a progressive contaminant accumulation likely associated with intensified summer anthropogenic activity. Biomarker responses revealed clear activation of oxidative stress, with tissue-specific patterns. The findings confirm the effectiveness of active biomonitoring and multibiomarker approach in assessing coastal water quality and provide valuable baseline data for the management of marine ecosystems. Full article

Environmental exposures can influence phenotypes across generations, yet the cellular routes by which somatic stress signals reach the germline remain poorly defined. piRNAs are attractive candidates for transgenerational epigenetic inheritance because they silence transposable elements, guide chromatin regulation, carry a stabilizing 3′ 2′-O-methyl modification, and participate in self-reinforcing amplification pathways, including ping-pong amplification in animals and RNA-dependent RNA polymerase (RdRP)-mediated secondary small-RNA amplification in systems such as C. elegans. This review examines evidence linking piRNAs, macrophage biology, and environmentally induced inheritance. We first summarize small-RNA inheritance in animals, plants, and ciliates, emphasizing C. elegans piRNA-triggered epigenetic memory and plant RNA-directed DNA methylation as parallel small-RNA-based inheritance systems. We then discuss emerging evidence that macrophage polarization states contain distinct piRNA signatures and release extracellular vesicles carrying non-coding RNAs. Finally, we revisit the Drosophila ectopic large bristle outgrowth (ELBO) phenotype as a possible example of macrophage-like hemocytes linking stress, tissue remodeling, and heritable morphological variation. We propose the macrophage-mediated morphological evolution (M3) model as a testable framework connecting environmental stress to transgenerational phenotypes. Full article

(1) Reverse transcription quantitative real-time PCR (RT-qPCR) requires reliable reference genes for accurate data normalization; however, optimal reference genes for the economically and ecologically valuable timber species Phoebe zhennan remain uncharacterized; (2) Here, we selected nine candidate reference genes derived from full-length transcriptome sequencing to evaluate their expression stability across abiotic (drought) and biotic (Colletotrichum fructicola infection) stresses. Transcript abundance was analyzed via RT-qPCR using four distinct algorithms (Delta Ct, geNorm, NormFinder, and BestKeeper), with RefFinder used to reconcile analytical discrepancies and generate a definitive consensus ranking; (3) Our analysis showed that expression stability is highly context-dependent: CYP20-1 and HSP70-1 were the most stable reference genes under drought stress, whereas Actin-101 and Actin constituted the optimal pair under disease stress. For cross-condition assessments, Actin-101 and β-Tubulin served as the most reliable baseline combination. Subsequent empirical validation quantifying stress-responsive transcripts demonstrated a significant positive correlation between RT-qPCR relative expression and corresponding RNA-seq data (drought: R = 0.80; disease: R = 0.76); (4) This study identifies and validates the first set of reference genes for P. zhennan, providing a foundation for accurate gene expression analysis in this species, which is crucial for understanding its response to environmental stresses. Full article